Aguilera, et al, 2008-Hybridization of intelligent techniques and ARIMA models for time series predi
HER-2在胃癌中的研究进展
HER-2在胃癌中的研究进展高福平【摘要】人类表皮生长因子受体2(HER-2)是一种原癌基因,其与多种肿瘤的发生、发展及预后关系密切,通过下游信号的转导参与肿瘤细胞增殖、凋亡、浸润及转移等的调控.HER-2的过表达与肿瘤患者不良总体预后相关.作为胃癌预后指标、治疗反应预测指标、治疗靶点,其可预测新辅助化疗的疗效,HER-2已成为国际肿瘤生物治疗的热点.该文就HER-2的生物学特征、作用机制、检测方法及与胃癌生物学行为关系的研究作一综述.【期刊名称】《医学综述》【年(卷),期】2014(020)005【总页数】3页(P816-818)【关键词】人类表皮生长因子受体2;原癌基因;胃癌【作者】高福平【作者单位】高淳县人民医院病理科,南京,211300【正文语种】中文【中图分类】R735.2胃癌是一类常见的消化系统的恶性肿瘤,其发病率仅次于肺癌、乳腺癌、大肠癌,位列第4[1]。
近年来研究表明,胃癌的发生、发展过程复杂,与癌基因激活、抑癌基因失活等因素有关[2]。
寻找与胃癌发生、发展及预后相关的生物学指标、检测癌基因及抑癌基因表达产物对胃癌的影响,有助于胃癌的早期发现及诊断,并为胃癌的靶向治疗提供方向。
原癌基因人类表皮生长因子受体2(human epidermalgrowth factor receptor-2,HER-2)是一种磷酸化受体蛋白,其在肿瘤中过度表达,并影响肿瘤细胞的增殖、分化以及浸润转移,其在肿瘤的发生、发展中起重要作用。
1 HER-2基因概述1.1 HER-2的结构特征 HER-2基因是一种原癌基因,为表皮生长因子受体(epidermal growth factor receptor,EGFR)家族成员之一,属于Ⅰ型受体酪氨酸激酶,它是一种膜结合蛋白。
HER-2定位于人类第17号染色体长臂(17q21),编码产物为185×103的跨膜糖蛋白。
HER-2第1~653个氨基酸为胞外区,第654~675个氨基酸为穿膜区,第676~1255个氨基酸为胞内区,其蛋白表达产物具有酪氨酸激酶活性的跨膜糖蛋白,通过细胞间的信号转导,调节细胞增殖、分化及生长。
20种甘蓝型油菜抗根肿病资源的筛选
20种甘蓝型油菜抗根肿病资源的筛选高愿;高峰;李淑娟;彭少丹;陈升位;林良斌【摘要】模拟根肿病发病条件,采用菌土接种法进行盆栽试验,对20个甘蓝型油菜品种进行根肿病抗性鉴定.根据病情指数将20个甘蓝型油菜品种的根肿病抗性分为高抗、抗病、低感、感病和高感.结果显示:华杂9号、华双3号的病情指数分别为25.06,25.03,属高抗品种;云花油早熟1号、陕油8号的病情指数分别为34.6,36.92,属抗病品种;中双9号、花油5号、花油8号的病情指数分别为45.8,46.89,47.33,属低感品种;玉红油2号等5个品种属感病品种;宁油16号等8个品种属高感品种.因此,华杂9号、华双3号表现为较强的抗根肿病能力,可以作为油菜抗根肿病育种的材料.【期刊名称】《云南农业大学学报》【年(卷),期】2015(030)003【总页数】5页(P346-350)【关键词】油菜;根肿病;抗性【作者】高愿;高峰;李淑娟;彭少丹;陈升位;林良斌【作者单位】云南农业大学农学与生物技术学院,云南昆明650201;云南农业大学农学与生物技术学院,云南昆明650201;云南农业大学农学与生物技术学院,云南昆明650201;云南农业大学农科专业实验教学中心,云南昆明650201;云南农业大学农学与生物技术学院,云南昆明650201;云南农业大学农学与生物技术学院,云南昆明650201【正文语种】中文【中图分类】S565.402.4芸薹根肿病会侵染和危害十字花科植物,给我国油菜和蔬菜生产造成了巨大损失[1-2]。
前人通过对芸薹属大白菜抗根肿病的遗传分析发现了Crr1,Crr2,Crr3,Crr4,CRa,CRb,CRc,CRk[3-9]8个抗根肿病(clubroot resistance,CR)基因;SAITO等[10]通过对拟南芥抗根肿病的QTL分析,精准定位了Crr3基因;HATAKEYAMA等[11]在大白菜中精准定位了Crr1基因;PIAO等[8-9]通过大白菜发现CRb基因对根肿病生理小种2,4,8表现为抗性;CHIAUG[12]发现在欧洲油菜中,抗生理小种1和抗生理小种3是受到单一显性基因控制,抗生理小种5是受2对隐性基因控制;GRUTE等[13]在研究了芸薹根肿病菌的某些菌株与3种芸薹属植物(芜菁、油菜、甘蓝)基因型的关系后,认为油菜和芜菁对不同菌株的抗性是不同的,它们可能是由寡基因控制的;郭海风[14]还发现抗病的油菜基因型和芜菁基因型之间可能存在共同的基因。
蛋白酶K的生理特性及应用(含文献列表)
特异性蛋白酶K具有广泛的底物特异性。
即便存在洗涤剂的情况下,其仍可降解许多非变性状态的蛋白质。
蛋白酶K分离自一种可在角质上生长的腐生真菌(Tritirachium album)。
因此,蛋白酶K能够降解非变性状态的角质(头生理性质蛋白酶K是稳定的S8家族丝氨酸碱性蛋白酶,在邻近活性位点组氨酸的位置含有两个二硫键和一个游离半抑制剂起到抑制作用。
蛋白酶K不会受到碘乙酸、胰蛋白酶特异性抑制剂TLCK、糜胰蛋白酶特异性抑制剂TPCK以及对-氯汞基苯甲酸盐抑制。
应用制备说明蛋白酶K可溶于水(1mg/ml),获得无色透明的溶液。
溶解性和溶液稳定性建议在-20℃下冻存粉末。
产品可稳定保持至少2年。
蛋白酶K溶液在较广的pH范围内(4.0-12.5,最适pH8.0)保持稳定,同时在使用时可在25-65℃范围内保持稳定。
pH8.0时,溶液至少可在4℃下稳定保存12个月[3]。
pH4-11.5时,含有Ca2+ (1-6mM)的溶液预计可稳定保存数周。
80%的硫酸铵悬液可在4℃下至少稳定保存12个月[2]。
文献列表[1]Betzel, C., Three Dimensional Structure of Proteinase K at 0.15 nm Resolution. Eur. J. Biochem., 178, 155-171 (1988).[2]Ebeling, W., et al., Proteinase K from Tritirachium album Linder, Eur. J. Biochem., 47, 91 (1974).[3]Enzymes of Molecular Biology, vol. 16, Burrell, M.M., ed. Humana Press (Totowa, NJ: 1993), p. 307. Kraus,E., and Femfert, U., Proteinase K from the Mold Tritirachium album Limber, Specificity and Mode of Action. Z. Physiol. Chem., 357, 937 (1976).[4]Lizardi, P.M., and Engelberg, A., Rapid Isolation of RNA Using Proteinase K and Sodium Perchlorate. Anal. Biochem., 98, 116 (1979).[5]Gross-Bellard, et al., Isolation of High Molecular Weight DNA from Mammalian Cells, Eur. J. Biochem., 36, 32-38 (1973).[6]Molecular Cloning: A Laboratory Handbook, 2nd ed., Sambrook et al., eds., Cold Spring Harbor Press (Cold Spring Harbor, NY: 1989) p. 1.61 and p. B.16.[7]Kasche, V., et al., A Two-step Procedure for Quantitative Isolation of Pure Double-strand DNA from Animal Tissues and Cell Cultures. Prep. Biochem., 11, 233 (1981).[8]Hansen, J.N., Isolation of Higher Molecular Weight DNA from Bacillus cereus T Using Proteinase K. Prep. Biochem., 4, 473 (1974).[9]Holm, C., et al., A Rapid, Efficient Method for Isolating DNA from Yeast. Gene, 42, 169 (1986).[10]La Claire, J.W., and Herrin, D.L., Co-isolation of High-Quality DNA and RNA from Coenocytic Green Algae. Plant Mol. Biol. Reporter, 15, 263 (1997).[11]Petsch, P., et al., Proteinase K Digestion of Proteins Improves Detection of Bacterial Endotoxins by the Limulus Amebocyte Assay: Application for Endotoxin removal from Cationic Proteins. Anal. Biochem., 259, 42 (1998).[12]Brdiczyka, D., and Krebs, W., Localization of Enzymes by Means of Proteases. Biochem. Biophys. Acta, 297, 203 (1973).[13]Short, B.G., et al., Automated Double Labeling of Proliferation and Apoptosis in Glutathione S-transferase-positive Hepatocytes in Rats. J. Histochemistry and Cytochemistry, 45, 1299 (1997).[14]Angerer, L.M., et al., Identification of Tissue-Specific Gene Expression by in-situ Hybridization. Methods in Enzymology, 152, 649 (1987).[15]Sakaguchi, S., et al., Accumulation of Proteinase K-Resistant Prion Protein (PrP) is Restricted by the Expression Level of Normal PrP in Mice Inoculated with a Mouse-Adapted strain of the Creutzfeldt-Jakob Disease Agent. J. Virology, 69, 7586 (1995).[16]Bennion, B.J., and Daggett, V., Protein Conformation and Diagnostic Tests: the Prion Protein. Clinical Chemistry, 48, 2105 (2002).[17]Hori, R., and Carey, M., Protease Footprinting Analysis of Ternary Complex Formation by Human TFIIA. J. Biol. Chem., 272, 1180 (1997).[18]Hilz, H., et al., Stimulation of Proteinase K action by Denaturing Agents: Application to the Isolation of Nucleic Acids and the degradation of “Masked” Proteins. Eur. J. Biochem., 56, 103 (1975).[19]Methods of Enzymatic Analysis, 3rd Edition, Bergmeyer, H.U., ed., Academic Press (New York, NY: 1983) vol. 2, p. 299.[20]Jany, K.D., et al., Amino Acid Sequence of Proteinase K from the Mold, Tritirachium album Linder. Proteinase K; a Subtilisin-related Enzyme with Disulfide Bonds. FEBS Letters, 199, 139 (1986).[21]Jany, K.D., and Mayer, B., Proteinase K from Tritirachium album linder, Molecular Mass and Sequence Around the Active Serine Residue. Biol. Chem. Hoppe-Seyler, 366, 485 (1985).[22]Bajorath, J., et al., The Enzymatic Efficiency of Proteinase K is Controlled by Calcium. Eur. J. Biochem., 176, 441-447 (1988).[23]IUBMB Enzyme Nomenclature: /iubmb/enzyme/EC3/4/21/64.html。
中国牧草育种中存在的问题
中国牧草育种中存在的问题孙进武,魏鹏飞,王跃栋甘肃农业大学草业学院,甘肃兰州(730070)Email:wykissme@摘要:本文综述了我国牧草种质资源、新品种培育、良种繁育等方面的取得的研究成绩,分析了我国牧草育种研究中存在的育种方法较为落后、优异牧草种质资源匮乏、良繁体系不健全等问题,从而提出了以现代生物技术为中心的解决建议.关键词:牧草育种,研究成绩,解决建议,生物技术1 中国牧草育种研究的成绩1.1 牧草种质资源方面新中国的成立标志新时代的到来,人类对各方面的需求日益增长,我国牧草育种也取得相当不错的成绩。
首先,初步完成了我国牧草品种资源的考察、收集、鉴定评价、入库保存。
现已查明我国牧草野生资源中至少有28科、184属、567种,共3296份材料具有保护、引种、育种价值。
国家牧草中期库已保存牧草种质3500多份,可保存20~ 25年。
另外,在我国不同气候带的生态区建立起5处多年生牧草种质资源圃,对一些材料的生物学特性和农艺性状等开展了鉴定和评价,并建立起了中国牧草与草地资源网站,通过以上工作,初步建立了以国家长期库、中期库为核心,多年生牧草种质资源圃为网络的保存体系[1]。
1.2 牧草常规育种上世纪50年代开始,中国的牧草育种工作者采用野生引种驯化、地方品种整理、国内外优良品种引进、选择育种及杂交育种等基本的育种方法培育出一批新品种,经全国牧草品种审定委员会审定登记的品种达250个,其中野生栽培品种40个、地方品种40个、引进品种6个、育成品种84个。
从国外引进包括苜蓿、三叶草、百脉根、多年生黑麦草、猫尾草、黄花草木樨、白花草木樨等种质,为新品种的培育奠定了一定基础。
大量引种试验筛选出一批适应我国不同地域的草种,如蔚县苜蓿、晋南苜蓿、柱花草、白三叶、多年生黑麦草等;并驯化了一批野生牧草,如羊草、老芒麦、无芒雀麦、黄花苜蓿、沙打旺、披碱草等。
人工选育种研究也有不少贡献,如吉林农业科学院通过系统选育培育出高产、抗寒、适应性广的公农1号、公农2号苜蓿新品种;内蒙古农业大学通过杂交培育出抗寒抗旱的草原1号、草原2号、草原3号杂花苜蓿;甘肃农业大学、新疆农业大学培育出了甘农1号、甘农2号、甘农3号、新牧1号、新牧2号、新牧3号苜蓿新品种等等[2]。
Small-2008-Microfluidic Synthesis of Nanomaterials
Microfluidics DOI:10.1002/smll.200701029Microfluidic Synthesis of NanomaterialsYujun Song,Josef Hormes,and Challa S.S.R.Kumar*From the Contents1.Introduction (699)2.Microfluidic Reactors:An Overview (700)3.Unique Features ofMicrofluidic Reactorsfor Controlled Synthesisof Nanomaterials (701)4.Discussion andAnalysis (706)5.Conclusion (707)Keywords:controlled synthesismicrofluidicsnanomaterialsnanoparticles698ß2008Wiley-VCH Verlag GmbH&Co.KGaA,Weinheim small2008,4,No.6,698–711A n overview of the current information and analyses on the microfluidic synthesis of different types of nanomaterial,including metallic and silica nanoparticles and quantum dots,is presented.Control of particle size,size distribution,and crystal structure of nanomaterials are examined in terms of the special features of microfluidic reactors.1.IntroductionMicrofluidic reactors,a part of the vastfield of micro-fabrication,deal with theflow of minute amounts of liquid within micrometer-size channels.Although microfluidic reactors werefirst developed in the early1990s,sta-te-of-the-art developments in recent years in the design, fabrication,and utilization of microfluidic devices have found many applications in pharmaceutical,biotechnology,and chemical industries for applications such asfine chemical synthesis,diagnosis,crystallization,combinatorial synthesis, rapid chemical analyses,and high-throughput screening.[1–3] In addition,several possibilities to fabricate three-dimensional (3D)networks of nanoscaleflow paths and the ability to focus nanoparticles within microfluidic systems are beginning to open up[4,5]and are likely to lead to unique microfluidic,even nanofluidic,[6]designs and reactors for a newer variety of applications.Therefore,it is not surprising that these developments in microfluidic devices coupled with nanotech-nology are beginning to pave the way for growing numbers of investigations to replace,in the future,conventional synthesis of nanomaterials by lab-on-a-chip systems.Microfluidic reactors offer many potential advantages in the chemical industry due to superior reaction control,high throughput and safer operational environment.[1,2]Complica-tions in traditional batch processes associated with large-scale transport and storage as well as safety and health issues(such as explosion and leakage of toxic andflammable solvents)are minimized.[7,8]Process scale-up,based on the concept of parallel processing,with a precise control of the synthetic aspects of thefinal product to produce chemicals with well-defined and pre-determined properties in higher yields, has been well demonstrated.[1,2,7–9]The option of manipulat-ing and controlling additional reaction parameters,compared to traditional batch processes,such asflow rates and the ability to cease the reaction as soon as the product is formed in a microliter or even nanoliter volume,offers even more possibilities for product control.[9]Fundamentally,it is pertinent to understand the various forces and interactions within the microfluidic environment that impact the outcome of a reaction.In addition to producing diffusive mixing,an inherently high surface-to-volume ratio enables heat generated by exothermic reactions to be dissipated rapidly,thereby creating more‘‘active sites’’for reactions.[10]The impact of surface forces(e.g.,surface tension),electrical effects,van der Waals interactions and surface roughness of the channel side walls become more significant and often dominate the particle motion,particularly for nanometer-sized materials(such as proteins,DNA,nanocrystals,etc.).[11]The possibility to create complicated 3Dfluidic geometries[4,5]offers unique opportunities to manipulate these surface forces as their effects on nanome-ter-sized materials are significant.In addition to the‘‘laminar flow-dominant’’microfluidic reactors,‘‘segmentedflow’’microfluidic reactors are providing alternate approaches to chemical syntheses.[3c,12]Through the possibility of introdu-cing time-control parameters(flow rates and sequence of reagent addition),reactions can be controlled at different stages,resulting in aflexible kinetic control and reactor design according to the reaction mechanism.The main advantage,as well as a challenge,in using microfluidic approaches is the possibility to creatively use these different effects and reaction parameters to produce tailor-made nanomaterials.While the recent publications clearly demonstrate the potential of microfluidic reactor technology in chemical syntheses,we believe that microfluidic reactors are likely to play an extremely important role in nanomaterials synthesis. Nanomaterials arefinding a number of unique applications in a broad range offields such as catalysis,biomedical, electronics,and the environment.[13]As they exhibit a variety of size-,shape-,and crystal-structure-dependent physical and chemical properties,their synthesis requires superior control of reactions,both kinetic and thermodynamic,in order to produce materials of required features and properties.[14]The role of microreactors in nanomaterials synthesis is therefore primarily in three areas:1)size-,shape-,and structure-controlled syntheses,2)scale-up through continuousflow processes,and3)high-throughput tools in process develop-ment.The Review presented here provides up-to-date information and analysis on microfluidic synthesis of nano-materials.It also compares the efficiency of microfluidic reactor processes with the conventional batch processes. Syntheses of different types of nanomaterial(nanoparticles, nanowires,nanotubes,etc.)using microfluidic reactors are also reviewed and attempts tofind explanations for the claimed control over particle size,size distribution,and crystal structure of nanomaterials,vis a vis special features of microfluidic reactors,have also been made.As results reported in the majority of the publications are more qualitative than quantitative,it is rather difficult to obtain a complete and clear picture about the potential of microfluidic rectors for nanomaterial synthesis.Nevertheless,we can draw [Ã] C.S.S.R.Kumar,Y.Song,J.HormesCenter for Advanced Microstructures and Devices at Louisiana State University6980Jefferson Hwy.,Baton Rouge,LA70806(USA)E-mail:ckumar1@small2008,4,No.6,698–711ß2008Wiley-VCH Verlag GmbH&Co.KGaA,Weinheim 699some valuable conclusions and lessons.In addition,weanticipate that the topic is of interest to a large group of scientists and this Review will serve as a guide,generate enthusiasm,and spark new developments in thefield of ‘‘microfluidic synthesis of nanomaterials’’.The Review is divided into three major sections.In thefirst section,an overview of microfluidic reactors is provided for those who are not familiar with microfluidic reactor technology.The second section contains an analysis of controlled synthesis of nanomaterials using some of the unique features that microfluidic reactors offer.Finally,the last section presents some important conclusions on the status,trends,and a future perspective of this new technology.2.Microfluidic Reactors:An OverviewThe fabrication of microfluidic reactors can be divided into four major stages:in thefirst stage,microstructuring of wafers and polymer substrates to form microchannels is carried out by various microfabrication processes.The microstructures, which may be endowed with functional surfaces,are designed to function as catalytic reaction channels or micromixers, microsensors,microseparators,and so on.In the second stage, microstructures are sealed and then bonded to a substrate by welding,gluing,or by using any other suitable bonding process.In the third stage,integration of various components such as microparts(mixers,actuators,pumps,separators,and heat exchanger)is carried out to form a microfluidic device.In thefinal stage,the microfluidic device is connected to the macroscale world to provide the microparts with energy and the chemicals necessary to carry out chemical synthesis,to collect the products and to analyze product/process develop-ment data,if necessary.More specific details on several types of microfabrication processes can be found in several review articles.[15]A microfluidic device,in general,needs to be integrated with various components such as micromixers,microscale heat exchanger,micropumps,microsensors such as pressure and flow sensors,microextractors,and microactuators depending on the nature of the reaction being carried out and the type of application.To date,fabricating a single microscale reactor chip with all the microcomponents integrated is still a challenge.Also,providingflexible connections with macro-scale feed and detector is still not a trivial ually,screws or orifice holes are designed in the substrate(e.g.,glass, polymers)itself and the tubing from the external devices(such as syringe pumps,product collectors)is connected by direct gluing to the opening parts of the microfluidic reactor through fittings and nuts.[10b,16–18]The microreactors used for nano-particle synthesis usually have micromixers,microchannels, and microheaters in a single reactor chip.[19,20–26] While the majority of studies on nanomaterial synthesis reports using microfluidic reactors are based on conventional laminarflow,other reactor designs such as tubular reactors, segmented-flow reactors,microstructured mixers,and micro-fluidic droplet reactors are also beginning to show promise.[27–29]Inherent problems associated with laminar-flow-based microfluidic reactors such as velocity and residenceDr.Yujun Song is Professor at the KeyState Laboratory of Chemical ResourceEngineering and College of MaterialsScience and Engineering at BeijingUniversity of Chemical Technology inthe area of integrated micro-andnanomaterials and devices.Heobtained his Ph.D.in Materials Scienceand Engineering from Beijing Universityof Chemical Technology(BUCT).Prior tojoining BUCT,Dr.Song was a post-doctoral researcher in the nanofabrication group at the Center for Advanced Microstructures and Devices(CAMD)working in such wide-ranging areas as microfluidic devices,nanoparticle and polymer composites,and nanoparticle synthesis and character-ization,and then worked at the Applied Research Center of Old Dominion University in the area of fabrication and functionalization of nanomaterials and their applications in biotechnology.Josef Hormes is Professor of Physics atLouisiana State University and theDirector of the Center for AdvancedMicrostructures and Devices(CAMD).Prior to joining CAMD in1999he wasfor about15years the Director of theSynchrotron Radiation Facility at BonnUniversity(Bonn,Germany).He wasalso a visiting professor at the ImperialCollege,London,and the Institute ofPhysics in Stockholm.He has beenactively involved in‘nano’research especially in application of X-ray absorption spectroscopic tools for characterization of nanomaterials.He has nearly200peer reviewed publications.He has a doctorate degree and a Habilitation in Physics from Bonn University.Challa Kumar obtained his doctoratedegree in synthetic organic chemistryfrom Sri Sathya Sai Institute of HigherLearning,Prashanti Nilayam,India.Heis currently the leader of the Nano-fabrication group at the Center forAdvanced Microstructures andDevices,Baton Rouge,USA.Hisresearch interests are in developingnovel synthetic methods for functionalnanomaterials with special emphasison microreactor technologies.He isactively involved with several collab-orators in developing innovative therapeutic,diagnostic,and sensor tools based on nanotechnology.He has seven years of industrial research and development experience working for ICI plc. and UB Ltd.He has worked at the Max Planck Institute fu¨r Biochemie,Munich as a post-doctoral fellow and at the Max Planck Institute fu¨r Kohlenforschung,Mulheim as an invitedscientist.ß2008Wiley-VCH Verlag GmbH&Co.KGaA,Weinheim small2008,4,No.6,698–711time distributions are to a certain extent addressed in segmented-flow microfluidic reactors,leading to better control over nanoparticle size distribution.However,the problem of physical contact of the particles with the walls of the reactor channels,resulting in cross contamination and significant volume changes due to gases utilized,still exists.One possible solution for these problems is to use microfluidic droplet reactors,where the precursor solutions can be encapsulated within nanoliter-scale droplets of the carrierfluid.Two microfluidic devices based on the concept of droplet reactors have been reported for the synthesis of nanoparticles.In the first device,as shown in Figure1,a polydimethylsiloxane (PDMS)-based reactor was fabricated with microfluidic channels approximately50m m high,50m m wide,and 24.5mm long based on rapid prototyping in PDMS.[27,30,31] In the second device,as shown in Figure2,a glass-based microfluidic reactor with a droplet jet injector was specifically fabricated for the high-temperature synthesis of quantum dots (Q dots).[29,32]In addition to the droplet generator,the microfluidic reactor was designed to include200-m m-wide heated serpentine channels,with semicircular turns intended for mixing,thermocouple wells,and inlets.[33a]An out-of-plane expansion with sharp increase in channel height at the beginning of the nozzle was designed for producing monodisperse emulsions.[32]The microfluidic droplet reactor can be used to manipulate droplets over a wide range offlow rates with the ability to form droplets at a low viscosity ratio of the two immiscible liquids at high capillary numbers and temperatures.Though the techniques for microfluidic device fabrication have seen some rapid developments,especially for fabrication of microstructures that serve as pipes,valves,mixers,pumps,and so on,there is a need for further investigation into further improving the fabrication processes,utilization of appropriate materials,and,most importantly,in integrating microproces-sing components related to the microfluidic reactor system (such as micropumps or electro-osmosis parts,microheaters, microseparators).The interested reader can refer to the text by O.Levenspiel for more information on designing continuous-flow reactors.[33b]3.Unique Features of Microfluidic Reactors for Controlled Synthesis of NanomaterialsNanoparticles exhibit a variety of size-,shape-,and crystal-structure-dependent physical and chemical proper-ties.[8,9,34–37]Synthesis of nanoparticles,therefore,requires superior control of reactions,both kinetic and thermodynamic, in order to produce materials of required features and properties.[9,34,35,37–40]Some of the challenges,particularly in the control of particle size,have been overcome using traditional wet-chemical methods.[41]For example,reactions using inverse micelles,[42–44]polymer templates,[45–47]meso-porous ceramic hosts,[48,49]and high-temperature hydroly-sis[36,50–55]have been employed to obtain nearly monodisperse nanoparticles.However,most of the traditional approaches usually require multipurification steps in order to obtain stable monodisperse nanoparticles.[55]Nanoparticles made by using physical techniques such as laser vaporization,[56,57]sputter-ing,[58a]metal evaporation,[59b]and grinding(ball milling)[60] usually produce‘‘naked’’nanoparticles.[46]These are easilyFigure1.Micrograph of a PDMS microfluidic device for performing droplet based synthesis of nanoparticles.Reproduced with permission from Reference[27].Figure2.Microreactor with droplet jet injector.a)Channel schematic showing dimensions,inlets(b),thermocouple wells(O),and bound-aries of Kapton heater(square brackets).b)Optical micrograph of droplet-injection cross section.Octadecene is injected in the top channel,while the PFPE is injected in the side channels.The narrowest point is160m m wide.c)Lateral‘‘D’’-shaped cross section of channel etched on the bottom wafer only.d)Cross section of ellipsoidal channel etched on both top and bottom wafers.e)Axial cross section showing the45-m m step up in channel height.Reproduced with permission from Reference[29].small2008,4,No.6,698–711ß2008Wiley-VCH Verlag GmbH&Co.KGaA,Weinheim 701oxidized and agglomerate intensively,thereby decreasing the post-synthesis processing ability for their practical applica-tions.[60]Small production rates and expensive equipment also limit the commercialization of these‘‘physical’’meth-ods.[9,34,35,37,61,62]Unlike the physical methods,most wet-chemical methods,involving liquid-phase synthesis,can provide larger amounts of nanoparticles conveniently coated with organic stabilizers to protect them from aggregation and, possibly,oxidation from air,in some cases,using relatively simple equipment,which is crucial for commercialization.[42] A majority of the liquid-phase reactions are usually performed at high temperature(%2008C),limiting the choice of solvents and reagents.Metal-salt reduction to obtain nanoparticles is widely used in liquid-phase synthesis.However,chemical contamination of thefinal product from reducing agents, solvents,and other reagents utilized in wet-chemical synthesis is unavoidable.[36,50–61]Since it is difficult to control the reaction conditions precisely on a large scale in a liquid-phase synthesis,there is a need for using microfluidic-reactor-based synthetic methods where a better control over reaction conditions is expected to provide monodisperse nanoparticles of a defined size,shape,and crystal structure without recourse to multipurification steps.[36,45,50,63]The six key challenges in the wet-chemical synthesis of nanomaterials are:1)Can we synthesize monodisperse nanoparticles of a desired size?2)Can we obtain nanomater-ials with satisfactory crystallinity and the desired crystal structure and composition,particularly for alloy nanoparti-cles?3)Can we synthesize nonspherical,that is,anisotropic nanomaterials?4)Can wefine tune interactions between the stabilizing ligands and the nanoparticle surface,not only for better compatibility in the required application system but also for controlling the properties?5)Can we ensure self-assembly of the nanoparticles on a desired substrate with desired geometrical patterns?6)Can we produce nanomaterials with desired properties on a large scale?Over the past couple of years,preliminary experiments using microfluidic reactors demonstrated that physical proper-ties of a variety of nanomaterials,such as quantum dots, nanoparticles,nanotubes,nanowires,and nanocomposites can befine tuned through control ofnanocrystal growth parameters andkinetics.[19–21,23–25,64–66]In addition to themajority of laminar-flow-based microflui-dic approaches,microfluidic droplet tech-niques for nanoparticle synthesis have alsobeen found to be promising.[27–29]Based onthese results from initial investigations,onecan anticipate that microreactors are likelyto provide cost-effective and environmen-tal friendly technologies for the rationaldesign and synthesis of nanoparticles.Also,process scale-up either by utilizingseveral thousands of microreactors inparallel or using continuous-flow reactionprotocols can provide several advantagesin large-scale production of nanoparti-cles.[8,10,11,61]Some of the unique featuresof microfluidic reactors are the ability to:–efficiently mix reagents,using appropriate mixers,on a short time scale,resulting in an homogeneous reaction environ-ment throughout–investigate the fundamentals of nanoparticle formation through spatial resolution–operate within continuousflow regimes allowing additional reagents to be added downstream as required–control properties of nanomaterials by controlling their formation at the desired nucleation or growth stage–continuously vary the composition of a reaction mixture by varying differential injection rates of the inlet channels–scale up the synthesis with controlled kinetic parameters These features endow microfluidic devices with the significant potential to resolve many of the current issues in the wet-chemical synthesis of nanomaterials.The development of microfluidic devices for the synthesis of nanomaterials is still in its infancy.There is yet to be a clear demonstration of their superiority over‘‘flask’’reactions. However,the available information from the literature(as described briefly in the following examples)indicates that there is a potential for obtaining better control in size,size distribution,crystal structure,and shape of nanomaterials on both small-and large-scale processes.[9,18,34,64]Given below are some examples from the literature that illustrate how the above-mentioned critical features of microfluidic reactors are being utilized for the controlled synthesis of nanomaterials.3.1.MicromixingSeveral approaches to offset the negative impact of laminarflow and the absence of turbulence in the micro-channels have been undertaken in order to increase the efficiency of mixing and thereby improve the monodispersity of synthesized nanoparticles.For example,Edel et al. demonstrated the synthesis of CdS nanoparticles in a continuous-flow microfluidic reactor with a micromixer based on the principle of distributive mixing(Figure3a).[19]In contrast to the nanoparticles synthesized in traditionalflask reactions(inset curve in Figure3),nanoparticles synthesized in the microfluidic reactor showed a sharper decline inFigure3.a)Typical micromixer fabricated in a glass/silicon/glass sandwich for CdS nano-particles;b)absorption spectra offluid streams exiting the micromixer chip subsequent tomixing of cadmium nitrate and sodium sulfide solution as a function of volumetricflow rate.Graph:absorption spectrum of nanoparticles produced by mixing bulk solutions(750m L).Reproduced with permission from References[19]and[26].ß2008Wiley-VCH Verlag GmbH&Co.KGaA,Weinheim small2008,4,No.6,698–711polydispersity tending towards monodispersity rather than a gradual increase in polydispersity.Over all,it appears that,just downsizing the reaction vessel from bulk to microliter volume is sufficient to improve monodispersity of the nanocrystallites. The nanoparticle aggregation on channel surfaces is a common occurrence within microreactors having simple T-mixer geometries.In order to overcome this,an innovative microreactor system with a radial interdigitated mixer,as shown in Figure4,was utilized for the synthesis of nanoparticles of gold stabilized by an adsorbed monolayer of a thiol(monolayer-protected clusters).[67]Efficient mixing in the microfluidic syntheses produced particles with improved monodispersity with the standard deviations of particle size range between0.6and0.9nm in comparison with those obtained from the bulk syntheses,which vary between1.3and 2.1nm.Size-controlled Pd nanoparticles were also demonstrated using a polymer-based microfluidic reactor with mixing elements.[9]The completely polymeric microreactor used in this study is shown in Figure5a.The microfluidic reactor, fabricated using SU-8on a polyetheretherketone(PEEK) substrate,was found to be tolerant of a variety of organic solvents.It hasfive parallel reaction channels on a single10Â10-cm2chip and theflow rate in the channels can be varied from120m L minÀ1to2400m L minÀ1.In order to ensure that the reagents mix rapidly(fast on the time scale of the reaction),[68,69]two four-way mixers(Figure5b)are incorpo-rated into the reactor.Two additional four-way mixers were included to enhance the mixing efficiency and to prevent back flow.Pd nanoparticles obtained from the microreactor have a smaller mean particle diameter of3.0nm,with a narrower size distribution,in comparison to those obtained from theflask process.[9]Apart from the examples given above,recent investiga-tions have also focused on alternate ways to improving mixing efficiency and control of the concentration of reagents through microfluidic droplet fusion techniques,both at room and higher temperatures.[27–29]3.2.Spatial Resolution of KineticsIn addition to the possibility to provide opportunities to control the properties of the nanomaterials produced within the microreactor,the microfluidic system provides a unique platform for investigating the fundamental reaction processes through spatially resolved analysis of nanoparticle formation within the channels.This was recently demonstrated in our ing a PMMA microreactor,cobalt nanoparticle formation was probed at three different positions using synchrotron-radiation-based X-ray absorption spectro-scopy.[70]CoK-edge XANES spectra recorded at three different positions of the microchannel together with reference spectra of the precursor and thefinal product collected at the end of microfluidic system show that time resolution of the reaction(in the order of milliseconds)is obtained by spatial resolution within the microreactor. Similarly,Sounart et al.have recently reported spatially resolved photoluminescence imaging and spectroscopy of Q dot formation within a microfluidic reactor.[71b]The results from their investigation have provided direct insight into the kinetics and mechanistic data on the Q dot formation (Figure6).The study also shows how a diffusion-controlled reaction environment forces the nanoparticles to nucleate under uniform conditions of negligible precursor concentra-tion in a narrow region at the centre of the channel.Figure4.a)Three-dimensional schematic of a radial interdigitated mixer.Each mixer is fabricated in three layers.In thefirst two layers, inputflows are directed to two circular bus channels which,in turn,split theflow into eight identicalfluid laminae and deliver reagent streams towards a central mixing chamber.Thefinal layer acts as a cap to enclose channels and as a guide for input and output capillaries.The output is from the centre of the uppermost layer.b)Photograph of the fabricated mixer.Microchannels arefilled with dye solutions to show different shadings for the different channels.Reproduced with per-mission from Reference[67].Figure5.a)Schematic of the microreactor:1)Orifices to Feed A reservoir,Ø1.6mm;2)Feed A(metal salt THF solution)inlet channels, 150m m wide and9.5mm long;3)orifices to Feed B(LiBH(C2H5)3THF solution)reservoir,Ø1.6mm;4)Feed B inlet channels,100m m wide and5.5mm long;5),Four-way mixers;6,8,9)reaction channels, 300m m wide and70mm long for(6),400m m wide and120mm long for (7),400m m wide and160mm long for(8);7)four-pole mixers,smallest poles,100m m wide;10)orifices to product collector,Ø1.6mm.b)Four-way mixer.c)Multipole mixer.Reproduced with permission from Reference[16].small2008,4,No.6,698–711ß2008Wiley-VCH Verlag GmbH&Co.KGaA,Weinheim 7033.3.Online Variation of Reactant CompositionWet-chemical synthesis of nanomaterials using microflui-dic reactors can also take advantage of yet another unique ability of the microfluidic reactor to operate within con-tinuous-flow regimes allowing additional reagents to be added downstream as required.Such a feature allows for pre and post treatment and multistep synthesis in a single continuous-flow regime.Shestopalov et al.[27]carried out a multistep synthesis of Q dots at room temperature using the microfluidic droplet reactor shown in Figure1.The advantage of using such a reactor is that the reaction can be controlled on a millisecond time scale.Such a segmented-flow microfluidic approach temporarily isolates the reactants from the channel walls minimizing or eliminating cross contamination.As can be seen from thefigure,aqueous streams of a mixture of CdCl2, mercaptopropionic acid(MPA),and Na2S are injected into the reactor through the left and right inlets,with NaOH solution infused through the middle aqueous inlet.Droplets are formed when the aqueous streamsflow into theflow of oil within5ms(Figure7).The reaction is allowed to take place for 75ms and then quenched using MPA to improve the monodispersity of the particles obtained.A comparison of the reactions,with and without the use of a microreactor indicates better control over particle size distribution when the microreactor was used.The ratio of CdCl2/MPA to Na2S had an effect on the particle size.A twenty fold increase in the ratio resulted in obtaining smaller CdS particles.A more startling finding is that when the reaction was quenched using Na2S instead of MPA,larger CdS particles with Na2S-rich shells were obtained.These observations logically led to the development of a microfluidic droplet reactor for synthesis of core/shell nanoparticles.When the reaction was quenched using Na2Se instead of Na2S,CdS core/CdSe shell Q dots were obtained(Figure8).Findings such as these illustrate yet another unique feature of the microfluidic device for synthesis of nanoparticles where,in a single step,core/shell nano-particles can be obtained.The core/shell nanoparticle synthesis using microreactors therefore has advantages such as the ability to control overcoat thickness,avoid secondaryFigure6.Continuousflow microfluidic reactor used for observation of CdS-Cys NC synthesis at the boundary between two laminarflowing streams.a)Sketch of microfluidic device(channel width exaggerated); w¼100mm,L¼2cm,depth2h¼20mm.b)Fluorescent plume of CdS NPs,25mM CdSO4,Cd:S¼1:1,Cys:Cd¼4:1.Reproduced with per-mission from Reference[71].Figure7.Two-step synthesis on chip with millisecond quenching yields CdS colloidal nanoparticles that are less disperse than those syn-thesized without millisecond quenching.a)A schematic diagram of the microfluidic network.b)UV/Vis spectra of nanoparticles synthesized on chip with millisecond quench(A),on chip without quench(B),and on the bench top(C).Reproduced with permission from Reference[27]. Figure8.Two-step synthesis of nanoparticles with various sizes and composition.a)A schematic diagram of the microfluidic network. b)UV/Vis spectra of four different types of nanoparticle.A)CdS nanoparticles synthesized using a20:1ratio of CdCl2to Na2S with thiol quench.B)CdS nanoparticles synthesized using a1:1ratio of CdCl2to Na2S with thiol quench.C)CdS nanoparticles synthesized using a1:1 ratio of CdCl2to Na2S with Na2S quench.D)CdS/CdSe core/shell nanoparticles synthesized using a1:1ratio of CdCl2to Na2S with Na2Se quench.Reproduced with permission from Reference[27].ß2008Wiley-VCH Verlag GmbH&Co.KGaA,Weinheim small2008,4,No.6,698–711。
2008年诺贝尔化学奖简介
3 典型应用
3.1 活细胞内基因表达及蛋白质-蛋白质相互作用 的光学成像与可视化
随着人类基因组计划的完成,蛋白质功能研究 已成为生命科学面临的最重要任务之一. 传统的生 化方法只能对蛋白质进行体外分析,这些离体研究 不能反映活细胞内蛋白质动力学. GFP 标记技术结 合多种光学成像技术很好地回答了上述问题. 其中 基 于 GFP 的 荧 光 共 振 能 量 转 移 (fluorescence resonance energy transfer,FRET)技术是应用最广泛
的 , [19] 它 可 实 时 动 态 监 测 蛋 白 质 时 空 动 力 学 . FRET 是 供 体 分 子 和 受 体 分 子 在 近 距 离 范 围 内 (<10 nm)所发生的一种非辐射式能量转移现象,目 前在生物和医学中被广泛应用的是 CFP(供体)和 YFP(受体). 国内也开展了基于 FRET 的蛋白质功 能研究,并在蛋白酶活性检测[20~23]和钙离子浓度[24] 方面取得了较好进展. FRET 除了可研究蛋白酶活 性[25]、蛋白质构象[26]、蛋白质磷酸化[27]和钙离子浓 度测定[28]等之外,蛋白质相互作用的 FRET 检测是 目前最受关注的焦点. 细胞内众多生命活动都是通 过蛋白质复合体来介导的,而蛋白质复合体则是多
美国公司法证券法历年经典论文列表
美国是世界上公司法、证券法研究最为发达的国家之一,在美国法学期刊(Law Review & Journals)上每年发表400多篇以公司法和证券法为主题的论文。
自1994年开始,美国的公司法学者每年会投票从中遴选出10篇左右重要的论文,重印于Corporate Practice Commentator,至2008年,已经评选了15年,计177篇论文入选。
以下是每年入选的论文列表:2008年(以第一作者姓名音序为序):1.Anabtawi, Iman and Lynn Stout. Fiduciary duties for activist shareholders. 60 Stan. L. Rev. 1255-1308 (2008).2.Brummer, Chris. Corporate law preemption in an age of global capital markets. 81 S. Cal. L. Rev. 1067-1114 (2008).3.Choi, Stephen and Marcel Kahan. The market penalty for mutual fund scandals. 87 B.U. L. Rev. 1021-1057 (2007).4.Choi, Stephen J. and Jill E. Fisch. On beyond CalPERS: Survey evidence on the developing role of public pension funds in corporate governance. 61 V and. L. Rev. 315-354 (2008).5.Cox, James D., Randall S. Thoma s and Lynn Bai. There are plaintiffs and…there are plaintiffs: An empirical analysis of securities class action settlements. 61 V and. L. Rev. 355-386 (2008).6.Henderson, M. Todd. Paying CEOs in bankruptcy: Executive compensation when agency costs are low. 101 Nw. U. L. Rev. 1543-1618 (2007).7.Hu, Henry T.C. and Bernard Black. Equity and debt decoupling and empty voting II: Importance and extensions. 156 U. Pa. L. Rev. 625-739 (2008).8.Kahan, Marcel and Edward Rock. The hanging chads of corporate voting. 96 Geo. L.J. 1227-1281 (2008).9.Strine, Leo E., Jr. Toward common sense and common ground? Reflections on the shared interests of managers and labor in a more rational system of corporate governance. 33 J. Corp. L. 1-20 (2007).10.Subramanian, Guhan. Go-shops vs. no-shops in private equity deals: Evidence and implications.63 Bus. Law. 729-760 (2008).2007年:1.Baker, Tom and Sean J. Griffith. The Missing Monitor in Corporate Governance: The Directors’ & Officers’ Liability Insurer. 95 Geo. L.J. 1795-1842 (2007).2.Bebchuk, Lucian A. The Myth of the Shareholder Franchise. 93 V a. L. Rev. 675-732 (2007).3.Choi, Stephen J. and Robert B. Thompson. Securities Litigation and Its Lawyers: Changes During the First Decade After the PSLRA. 106 Colum. L. Rev. 1489-1533 (2006).4.Coffee, John C., Jr. Reforming the Securities Class Action: An Essay on Deterrence and Its Implementation. 106 Colum. L. Rev. 1534-1586 (2006).5.Cox, James D. and Randall S. Thomas. Does the Plaintiff Matter? An Empirical Analysis of Lead Plaintiffs in Securities Class Actions. 106 Colum. L. Rev. 1587-1640 (2006).6.Eisenberg, Theodore and Geoffrey Miller. Ex Ante Choice of Law and Forum: An Empirical Analysis of Corporate Merger Agreements. 59 V and. L. Rev. 1975-2013 (2006).7.Gordon, Jeffrey N. The Rise of Independent Directors in the United States, 1950-2005: Of Shareholder V alue and Stock Market Prices. 59 Stan. L. Rev. 1465-1568 (2007).8.Kahan, Marcel and Edward B. Rock. Hedge Funds in Corporate Governance and Corporate Control. 155 U. Pa. L. Rev. 1021-1093 (2007).ngevoort, Donald C. The Social Construction of Sarbanes-Oxley. 105 Mich. L. Rev. 1817-1855 (2007).10.Roe, Mark J. Legal Origins, Politics, and Modern Stock Markets. 120 Harv. L. Rev. 460-527 (2006).11.Subramanian, Guhan. Post-Siliconix Freeze-outs: Theory and Evidence. 36 J. Legal Stud. 1-26 (2007). (NOTE: This is an earlier working draft. The published article is not freely available, and at SLW we generally respect the intellectual property rights of others.)2006年:1.Bainbridge, Stephen M. Director Primacy and Shareholder Disempowerment. 119 Harv. L. Rev. 1735-1758 (2006).2.Bebchuk, Lucian A. Letting Shareholders Set the Rules. 119 Harv. L. Rev. 1784-1813 (2006).3.Black, Bernard, Brian Cheffins and Michael Klausner. Outside Director Liability. 58 Stan. L. Rev. 1055-1159 (2006).4.Choi, Stephen J., Jill E. Fisch and A.C. Pritchard. Do Institutions Matter? The Impact of the Lead Plaintiff Provision of the Private Securities Litigation Reform Act. 835.Cox, James D. and Randall S. Thomas. Letting Billions Slip Through Y our Fingers: Empirical Evidence and Legal Implications of the Failure of Financial Institutions to Participate in Securities Class Action Settlements. 58 Stan. L. Rev. 411-454 (2005).6.Gilson, Ronald J. Controlling Shareholders and Corporate Governance: Complicating the Comparative Taxonomy. 119 Harv. L. Rev. 1641-1679 (2006).7.Goshen , Zohar and Gideon Parchomovsky. The Essential Role of Securities Regulation. 55 Duke L.J. 711-782 (2006).8.Hansmann, Henry, Reinier Kraakman and Richard Squire. Law and the Rise of the Firm. 119 Harv. L. Rev. 1333-1403 (2006).9.Hu, Henry T. C. and Bernard Black. Empty V oting and Hidden (Morphable) Ownership: Taxonomy, Implications, and Reforms. 61 Bus. Law. 1011-1070 (2006).10.Kahan, Marcel. The Demand for Corporate Law: Statutory Flexibility, Judicial Quality, or Takeover Protection? 22 J. L. Econ. & Org. 340-365 (2006).11.Kahan, Marcel and Edward Rock. Symbiotic Federalism and the Structure of Corporate Law.58 V and. L. Rev. 1573-1622 (2005).12.Smith, D. Gordon. The Exit Structure of V enture Capital. 53 UCLA L. Rev. 315-356 (2005).2005年:1.Bebchuk, Lucian Arye. The case for increasing shareholder power. 118 Harv. L. Rev. 833-914 (2005).2.Bratton, William W. The new dividend puzzle. 93 Geo. L.J. 845-895 (2005).3.Elhauge, Einer. Sacrificing corporate profits in the public interest. 80 N.Y.U. L. Rev. 733-869 (2005).4.Johnson, . Corporate officers and the business judgment rule. 60 Bus. Law. 439-469 (2005).haupt, Curtis J. In the shadow of Delaware? The rise of hostile takeovers in Japan. 105 Colum. L. Rev. 2171-2216 (2005).6.Ribstein, Larry E. Are partners fiduciaries? 2005 U. Ill. L. Rev. 209-251.7.Roe, Mark J. Delaware?s politics. 118 Harv. L. Rev. 2491-2543 (2005).8.Romano, Roberta. The Sarbanes-Oxley Act and the making of quack corporate governance. 114 Y ale L.J. 1521-1611 (2005).9.Subramanian, Guhan. Fixing freezeouts. 115 Y ale L.J. 2-70 (2005).10.Thompson, Robert B. and Randall S. Thomas. The public and private faces of derivative lawsuits. 57 V and. L. Rev. 1747-1793 (2004).11.Weiss, Elliott J. and J. White. File early, then free ride: How Delaware law (mis)shapes shareholder class actions. 57 V and. L. Rev. 1797-1881 (2004).2004年:1Arlen, Jennifer and Eric Talley. Unregulable defenses and the perils of shareholder choice. 152 U. Pa. L. Rev. 577-666 (2003).2.Bainbridge, Stephen M. The business judgment rule as abstention doctrine. 57 V and. L. Rev. 83-130 (2004).3.Bebchuk, Lucian Arye and Alma Cohen. Firms' decisions where to incorporate. 46 J.L. & Econ. 383-425 (2003).4.Blair, Margaret M. Locking in capital: what corporate law achieved for business organizers in the nineteenth century. 51 UCLA L. Rev. 387-455 (2003).5.Gilson, Ronald J. and Jeffrey N. Gordon. Controlling shareholders. 152 U. Pa. L. Rev. 785-843 (2003).6.Roe, Mark J. Delaware 's competition. 117 Harv. L. Rev. 588-646 (2003).7.Sale, Hillary A. Delaware 's good faith. 89 Cornell L. Rev. 456-495 (2004).8.Stout, Lynn A. The mechanisms of market inefficiency: an introduction to the new finance. 28 J. Corp. L. 635-669 (2003).9.Subramanian, Guhan. Bargaining in the shadow of takeover defenses. 113 Y ale L.J. 621-686 (2003).10.Subramanian, Guhan. The disappearing Delaware effect. 20 J.L. Econ. & Org. 32-59 (2004)11.Thompson, Robert B. and Randall S. Thomas. The new look of shareholder litigation: acquisition-oriented class actions. 57 V and. L. Rev. 133-209 (2004).2003年:1.A yres, Ian and Stephen Choi. Internalizing outsider trading. 101 Mich. L. Rev. 313-408 (2002).2.Bainbridge, Stephen M. Director primacy: The means and ends of corporate governance. 97 Nw. U. L. Rev. 547-606 (2003).3.Bebchuk, Lucian, Alma Cohen and Allen Ferrell. Does the evidence favor state competition in corporate law? 90 Cal. L. Rev. 1775-1821 (2002).4.Bebchuk, Lucian Arye, John C. Coates IV and Guhan Subramanian. The Powerful Antitakeover Force of Staggered Boards: Further findings and a reply to symposium participants. 55 Stan. L. Rev. 885-917 (2002).5.Choi, Stephen J. and Jill E. Fisch. How to fix Wall Street: A voucher financing proposal for securities intermediaries. 113 Y ale L.J. 269-346 (2003).6.Daines, Robert. The incorporation choices of IPO firms. 77 N.Y.U. L. Rev.1559-1611 (2002).7.Gilson, Ronald J. and David M. Schizer. Understanding venture capital structure: A taxexplanation for convertible preferred stock. 116 Harv. L. Rev. 874-916 (2003).8.Kahan, Marcel and Ehud Kamar. The myth of state competition in corporate law. 55 Stan. L. Rev. 679-749 (2002).ngevoort, Donald C. Taming the animal spirits of the stock markets: A behavioral approach to securities regulation. 97 Nw. U. L. Rev. 135-188 (2002).10.Pritchard, A.C. Justice Lewis F. Powell, Jr., and the counterrevolution in the federal securities laws. 52 Duke L.J. 841-949 (2003).11.Thompson, Robert B. and Hillary A. Sale. Securities fraud as corporate governance: Reflections upon federalism. 56 V and. L. Rev. 859-910 (2003).2002年:1.Allen, William T., Jack B. Jacobs and Leo E. Strine, Jr. Function over Form: A Reassessment of Standards of Review in Delaware Corporation Law. 26 Del. J. Corp. L. 859-895 (2001) and 56 Bus. Law. 1287 (2001).2.A yres, Ian and Joe Bankman. Substitutes for Insider Trading. 54 Stan. L. Rev. 235-254 (2001).3.Bebchuk, Lucian Arye, Jesse M. Fried and David I. Walker. Managerial Power and Rent Extraction in the Design of Executive Compensation. 69 U. Chi. L. Rev. 751-846 (2002).4.Bebchuk, Lucian Arye, John C. Coates IV and Guhan Subramanian. The Powerful Antitakeover Force of Staggered Boards: Theory, Evidence, and Policy. 54 Stan. L. Rev. 887-951 (2002).5.Black, Bernard and Reinier Kraakman. Delaware’s Takeover Law: The Uncertain Search for Hidden V alue. 96 Nw. U. L. Rev. 521-566 (2002).6.Bratton, William M. Enron and the Dark Side of Shareholder V alue. 76 Tul. L. Rev. 1275-1361 (2002).7.Coates, John C. IV. Explaining V ariation in Takeover Defenses: Blame the Lawyers. 89 Cal. L. Rev. 1301-1421 (2001).8.Kahan, Marcel and Edward B. Rock. How I Learned to Stop Worrying and Love the Pill: Adaptive Responses to Takeover Law. 69 U. Chi. L. Rev. 871-915 (2002).9.Kahan, Marcel. Rethinking Corporate Bonds: The Trade-off Between Individual and Collective Rights. 77 N.Y.U. L. Rev. 1040-1089 (2002).10.Roe, Mark J. Corporate Law’s Limits. 31 J. Legal Stud. 233-271 (2002).11.Thompson, Robert B. and D. Gordon Smith. Toward a New Theory of the Shareholder Role: "Sacred Space" in Corporate Takeovers. 80 Tex. L. Rev. 261-326 (2001).2001年:1.Black, Bernard S. The legal and institutional preconditions for strong securities markets. 48 UCLA L. Rev. 781-855 (2001).2.Coates, John C. IV. Takeover defenses in the shadow of the pill: a critique of the scientific evidence. 79 Tex. L. Rev. 271-382 (2000).3.Coates, John C. IV and Guhan Subramanian. A buy-side model of M&A lockups: theory and evidence. 53 Stan. L. Rev. 307-396 (2000).4.Coffee, John C., Jr. The rise of dispersed ownership: the roles of law and the state in the separation of ownership and control. 111 Y ale L.J. 1-82 (2001).5.Choi, Stephen J. The unfounded fear of Regulation S: empirical evidence on offshore securities offerings. 50 Duke L.J. 663-751 (2000).6.Daines, Robert and Michael Klausner. Do IPO charters maximize firm value? Antitakeover protection in IPOs. 17 J.L. Econ. & Org. 83-120 (2001).7.Hansmann, Henry and Reinier Kraakman. The essential role of organizational law. 110 Y ale L.J. 387-440 (2000).ngevoort, Donald C. The human nature of corporate boards: law, norms, and the unintended consequences of independence and accountability. 89 Geo. L.J. 797-832 (2001).9.Mahoney, Paul G. The political economy of the Securities Act of 1933. 30 J. Legal Stud. 1-31 (2001).10.Roe, Mark J. Political preconditions to separating ownership from corporate control. 53 Stan. L. Rev. 539-606 (2000).11.Romano, Roberta. Less is more: making institutional investor activism a valuable mechanism of corporate governance. 18 Y ale J. on Reg. 174-251 (2001).2000年:1.Bratton, William W. and Joseph A. McCahery. Comparative Corporate Governance and the Theory of the Firm: The Case Against Global Cross Reference. 38 Colum. J. Transnat’l L. 213-297 (1999).2.Coates, John C. IV. Empirical Evidence on Structural Takeover Defenses: Where Do We Stand?54 U. Miami L. Rev. 783-797 (2000).3.Coffee, John C., Jr. Privatization and Corporate Governance: The Lessons from Securities Market Failure. 25 J. Corp. L. 1-39 (1999).4.Fisch, Jill E. The Peculiar Role of the Delaware Courts in the Competition for Corporate Charters. 68 U. Cin. L. Rev. 1061-1100 (2000).5.Fox, Merritt B. Retained Mandatory Securities Disclosure: Why Issuer Choice Is Not Investor Empowerment. 85 V a. L. Rev. 1335-1419 (1999).6.Fried, Jesse M. Insider Signaling and Insider Trading with Repurchase Tender Offers. 67 U. Chi. L. Rev. 421-477 (2000).7.Gulati, G. Mitu, William A. Klein and Eric M. Zolt. Connected Contracts. 47 UCLA L. Rev. 887-948 (2000).8.Hu, Henry T.C. Faith and Magic: Investor Beliefs and Government Neutrality. 78 Tex. L. Rev. 777-884 (2000).9.Moll, Douglas K. Shareholder Oppression in Close Corporations: The Unanswered Question of Perspective. 53 V and. L. Rev. 749-827 (2000).10.Schizer, David M. Executives and Hedging: The Fragile Legal Foundation of Incentive Compatibility. 100 Colum. L. Rev. 440-504 (2000).11.Smith, Thomas A. The Efficient Norm for Corporate Law: A Neotraditional Interpretation of Fiduciary Duty. 98 Mich. L. Rev. 214-268 (1999).12.Thomas, Randall S. and Kenneth J. Martin. The Determinants of Shareholder V oting on Stock Option Plans. 35 Wake Forest L. Rev. 31-81 (2000).13.Thompson, Robert B. Preemption and Federalism in Corporate Governance: Protecting Shareholder Rights to V ote, Sell, and Sue. 62 Law & Contemp. Probs. 215-242 (1999).1999年(以第一作者姓名音序为序):1.Bankman, Joseph and Ronald J. Gilson. Why Start-ups? 51 Stan. L. Rev. 289-308 (1999).2.Bhagat, Sanjai and Bernard Black. The Uncertain Relationship Between Board Composition and Firm Performance. 54 Bus. Law. 921-963 (1999).3.Blair, Margaret M. and Lynn A. Stout. A Team Production Theory of Corporate Law. 85 V a. L. Rev. 247-328 (1999).4.Coates, John C., IV. “Fair V alue” As an A voidable Rule of Corporate Law: Minority Discounts in Conflict Transactions. 147 U. Pa. L. Rev. 1251-1359 (1999).5.Coffee, John C., Jr. The Future as History: The Prospects for Global Convergence in Corporate Governance and Its Implications. 93 Nw. U. L. Rev. 641-707 (1999).6.Eisenberg, Melvin A. Corporate Law and Social Norms. 99 Colum. L. Rev. 1253-1292 (1999).7.Hamermesh, Lawrence A. Corporate Democracy and Stockholder-Adopted By-laws: Taking Back the Street? 73 Tul. L. Rev. 409-495 (1998).8.Krawiec, Kimberly D. Derivatives, Corporate Hedging, and Shareholder Wealth: Modigliani-Miller Forty Y ears Later. 1998 U. Ill. L. Rev. 1039-1104.ngevoort, Donald C. Rereading Cady, Roberts: The Ideology and Practice of Insider Trading Regulation. 99 Colum. L. Rev. 1319-1343 (1999).ngevoort, Donald C. Half-Truths: Protecting Mistaken Inferences By Investors and Others.52 Stan. L. Rev. 87-125 (1999).11.Talley, Eric. Turning Servile Opportunities to Gold: A Strategic Analysis of the Corporate Opportunities Doctrine. 108 Y ale L.J. 277-375 (1998).12.Williams, Cynthia A. The Securities and Exchange Commission and Corporate Social Transparency. 112 Harv. L. Rev. 1197-1311 (1999).1998年:1.Carney, William J., The Production of Corporate Law, 71 S. Cal. L. Rev. 715-780 (1998).2.Choi, Stephen, Market Lessons for Gatekeepers, 92 Nw. U. L. Rev. 916-966 (1998).3.Coffee, John C., Jr., Brave New World?: The Impact(s) of the Internet on Modern Securities Regulation. 52 Bus. Law. 1195-1233 (1997).ngevoort, Donald C., Organized Illusions: A Behavioral Theory of Why Corporations Mislead Stock Market Investors (and Cause Other Social Harms). 146 U. Pa. L. Rev. 101-172 (1997).ngevoort, Donald C., The Epistemology of Corporate-Securities Lawyering: Beliefs, Biases and Organizational Behavior. 63 Brook. L. Rev. 629-676 (1997).6.Mann, Ronald J. The Role of Secured Credit in Small-Business Lending. 86 Geo. L.J. 1-44 (1997).haupt, Curtis J., Property Rights in Firms. 84 V a. L. Rev. 1145-1194 (1998).8.Rock, Edward B., Saints and Sinners: How Does Delaware Corporate Law Work? 44 UCLA L. Rev. 1009-1107 (1997).9.Romano, Roberta, Empowering Investors: A Market Approach to Securities Regulation. 107 Y ale L.J. 2359-2430 (1998).10.Schwab, Stewart J. and Randall S. Thomas, Realigning Corporate Governance: Shareholder Activism by Labor Unions. 96 Mich. L. Rev. 1018-1094 (1998).11.Skeel, David A., Jr., An Evolutionary Theory of Corporate Law and Corporate Bankruptcy. 51 V and. L. Rev. 1325-1398 (1998).12.Thomas, Randall S. and Martin, Kenneth J., Should Labor Be Allowed to Make Shareholder Proposals? 73 Wash. L. Rev. 41-80 (1998).1997年:1.Alexander, Janet Cooper, Rethinking Damages in Securities Class Actions, 48 Stan. L. Rev. 1487-1537 (1996).2.Arlen, Jennifer and Kraakman, Reinier, Controlling Corporate Misconduct: An Analysis of Corporate Liability Regimes, 72 N.Y.U. L. Rev. 687-779 (1997).3.Brudney, Victor, Contract and Fiduciary Duty in Corporate Law, 38 B.C. L. Rev. 595-665 (1997).4.Carney, William J., The Political Economy of Competition for Corporate Charters, 26 J. Legal Stud. 303-329 (1997).5.Choi, Stephen J., Company Registration: Toward a Status-Based Antifraud Regime, 64 U. Chi. L. Rev. 567-651 (1997).6.Fox, Merritt B., Securities Disclosure in a Globalizing Market: Who Should Regulate Whom. 95 Mich. L. Rev. 2498-2632 (1997).7.Kahan, Marcel and Klausner, Michael, Lockups and the Market for Corporate Control, 48 Stan. L. Rev. 1539-1571 (1996).8.Mahoney, Paul G., The Exchange as Regulator, 83 V a. L. Rev. 1453-1500 (1997).haupt, Curtis J., The Market for Innovation in the United States and Japan: V enture Capital and the Comparative Corporate Governance Debate, 91 Nw. U.L. Rev. 865-898 (1997).10.Skeel, David A., Jr., The Unanimity Norm in Delaware Corporate Law, 83 V a. L. Rev. 127-175 (1997).1996年:1.Black, Bernard and Reinier Kraakman A Self-Enforcing Model of Corporate Law, 109 Harv. L. Rev. 1911 (1996)2.Gilson, Ronald J. Corporate Governance and Economic Efficiency: When Do Institutions Matter?, 74 Wash. U. L.Q. 327 (1996)3. Hu, Henry T.C. Hedging Expectations: "Derivative Reality" and the Law and Finance of the Corporate Objective, 21 J. Corp. L. 3 (1995)4.Kahan, Marcel & Michael Klausner Path Dependence in Corporate Contracting: Increasing Returns, Herd Behavior and Cognitive Biases, 74 Wash. U. L.Q. 347 (1996)5.Kitch, Edmund W. The Theory and Practice of Securities Disclosure, 61 Brooklyn L. Rev. 763 (1995)ngevoort, Donald C. Selling Hope, Selling Risk: Some Lessons for Law From Behavioral Economics About Stockbrokers and Sophisticated Customers, 84 Cal. L. Rev. 627 (1996)7.Lin, Laura The Effectiveness of Outside Directors as a Corporate Governance Mechanism: Theories and Evidence, 90 Nw. U.L. Rev. 898 (1996)lstein, Ira M. The Professional Board, 50 Bus. Law 1427 (1995)9.Thompson, Robert B. Exit, Liquidity, and Majority Rule: Appraisal's Role in Corporate Law, 84 Geo. L.J. 1 (1995)10.Triantis, George G. and Daniels, Ronald J. The Role of Debt in Interactive Corporate Governance. 83 Cal. L. Rev. 1073 (1995)1995年:公司法:1.Arlen, Jennifer and Deborah M. Weiss A Political Theory of Corporate Taxation,. 105 Y ale L.J. 325-391 (1995).2.Elson, Charles M. The Duty of Care, Compensation, and Stock Ownership, 63 U. Cin. L. Rev. 649 (1995).3.Hu, Henry T.C. Heeding Expectations: "Derivative Reality" and the Law and Finance of the Corporate Objective, 73 Tex. L. Rev. 985-1040 (1995).4.Kahan, Marcel The Qualified Case Against Mandatory Terms in Bonds, 89 Nw. U.L. Rev. 565-622 (1995).5.Klausner, Michael Corporations, Corporate Law, and Networks of Contracts, 81 V a. L. Rev. 757-852 (1995).6.Mitchell, Lawrence E. Cooperation and Constraint in the Modern Corporation: An Inquiry Into the Causes of Corporate Immorality, 73 Tex. L. Rev. 477-537 (1995).7.Siegel, Mary Back to the Future: Appraisal Rights in the Twenty-First Century, 32 Harv. J. on Legis. 79-143 (1995).证券法:1.Grundfest, Joseph A. Why Disimply? 108 Harv. L. Rev. 727-747 (1995).2.Lev, Baruch and Meiring de V illiers Stock Price Crashes and 10b-5 Damages: A Legal Economic, and Policy Analysis, 47 Stan. L. Rev. 7-37 (1994).3.Mahoney, Paul G. Mandatory Disclosure as a Solution to Agency Problems, 62 U. Chi. L. Rev. 1047-1112 (1995).4.Seligman, Joel The Merits Do Matter, 108 Harv. L. Rev. 438 (1994).5.Seligman, Joel The Obsolescence of Wall Street: A Contextual Approach to the Evolving Structure of Federal Securities Regulation, 93 Mich. L. Rev. 649-702 (1995).6.Stout, Lynn A. Are Stock Markets Costly Casinos? Disagreement, Mark Failure, and Securities Regulation, 81 V a. L. Rev. 611 (1995).7.Weiss, Elliott J. and John S. Beckerman Let the Money Do the Monitoring: How Institutional Investors Can Reduce Agency Costs in Securities Class Actions, 104 Y ale L.J. 2053-2127 (1995).1994年:公司法:1.Fraidin, Stephen and Hanson, Jon D. Toward Unlocking Lockups, 103 Y ale L.J. 1739-1834 (1994)2.Gordon, Jeffrey N. Institutions as Relational Investors: A New Look at Cumulative V oting, 94 Colum. L. Rev. 124-192 (1994)3.Karpoff, Jonathan M., and Lott, John R., Jr. The Reputational Penalty Firms Bear From Committing Criminal Fraud, 36 J.L. & Econ. 757-802 (1993)4.Kraakman, Reiner, Park, Hyun, and Shavell, Steven When Are Shareholder Suits in Shareholder Interests?, 82 Geo. L.J. 1733-1775 (1994)5.Mitchell, Lawrence E. Fairness and Trust in Corporate Law, 43 Duke L.J. 425- 491 (1993)6.Oesterle, Dale A. and Palmiter, Alan R. Judicial Schizophrenia in Shareholder V oting Cases, 79 Iowa L. Rev. 485-583 (1994)7. Pound, John The Rise of the Political Model of Corporate Governance and Corporate Control, 68 N.Y.U. L. Rev. 1003-1071 (1993)8.Skeel, David A., Jr. Rethinking the Line Between Corporate Law and Corporate Bankruptcy, 72 Tex. L. Rev. 471-557 (1994)9.Thompson, Robert B. Unpacking Limited Liability: Direct and V icarious Liability of Corporate Participants for Torts of the Enterprise, 47 V and. L. Rev. 1-41 (1994)证券法:1.Alexander, Janet Cooper The V alue of Bad News in Securities Class Actions, 41 UCLA L.Rev. 1421-1469 (1994)2.Bainbridge, Stephen M. Insider Trading Under the Restatement of the Law Governing Lawyers, 19 J. Corp. L. 1-40 (1993)3.Black, Bernard S. and Coffee, John C. Jr. Hail Britannia?: Institutional Investor Behavior Under Limited Regulation, 92 Mich. L. Rev. 1997-2087 (1994)4.Booth, Richard A. The Efficient Market, portfolio Theory, and the Downward Sloping Demand Hypothesis, 68 N.Y.U. L. Rev. 1187-1212 (1993)5.Coffee, John C., Jr. The SEC and the Institutional Investor: A Half-Time Report, 15 Cardozo L. Rev 837-907 (1994)6.Fox, Merritt B. Insider Trading Deterrence V ersus Managerial Incentives: A Unified Theory of Section 16(b), 92 Mich. L. Rev. 2088-2203 (1994)7.Grundfest, Joseph A. Disimplying Private Rights of Action Under the Federal Securities Laws: The Commission's Authority, 107 Harv. L. Rev. 961-1024 (1994)8.Macey, Jonathan R. Administrative Agency Obsolescence and Interest Group Formation: A Case Study of the SEC at Sixty, 15 Cardozo L. Rev. 909-949 (1994)9.Rock, Edward B. Controlling the Dark Side of Relational Investing, 15 Cardozo L. Rev. 987-1031 (1994)。
解释了 HIV 入侵宿主细胞的一些途径
HIV vaccines and microbicides hold promise for prevent‑ing the acquisition of HIV‑1 and HIV‑2, the two viruses that cause AIDS, but the success of designing such agents needs a clear understanding of where HIV first encoun‑ters its target cells — primarily T cells, macrophages and dendritic cells (DCs) — and how it gains entry at various sites to eventually establish infection. HIV infection has rapidly spread since the early 1980s to become an epi‑demic disease (see the UNAIDS/WHO AIDS epidemic update) that is largely maintained by sexual transmis‑sion through the lower genital and rectal mucosa (FIG. 1, TABLE 1). Here, we have endeavoured to assemble the current knowledge on the acquisition of HIV at mucosal sites, confining our discussion to the lower genital mucosa. We clearly recognize that other sites of entry, such as the blood, placenta and gastrointestinal mucosa (BOX 1), are also important for HIV acquisition but these are beyond the scope of this Review.Many studies have provided insights into certain aspects of HIV and simian immunodeficiency virus (SIV) mucosal entry by carrying out detailed examinations of the relevant tissues and target cells following their in vivo, ex vivo or in vitro exposure to the virus. This Review discusses experimental systems that use the same mucosal source to avoid confusion and the inconsistencies that often emerge when findings from one system are extrapolated to those of another. Where appropriate, we have emphasized the benefits and limitations of the experimental approaches, important considerations in the interpretation of findings and their relevance for future studies.HIV invasion in the female genital tract Anatomical sites. An estimated 30–40% of all new HIV‑1 infections in women occur through vaginal intercourse, which carries a lower HIV transmission probability per exposure event than anal intercourse or parenteral inoculation (TABLE 1). Although HIV‑1 can infect the vaginal, ectocervical and endocervical mucosa (FIG. 1), the relative contribution of each site to the establish‑ment of the initial infection is not known. The multi‑layered squamous epithelium that covers the vagina and ectocervix, when intact, provides better mechani‑cal protection against HIV invasion than the single‑layer columnar epithelium that lines the endocervix. However, the greater surface area of the vaginal wall and ectocervix, which often exceeds 15 times that of the endocervix, provides more potential access sites for HIV entry, particularly when breaches occur in the epithelial‑cell layer. HIV or SIV can establish an initial infection solely through invasion of the vaginal mucosa, as shown in women who lack a uterus at birth1 and in female macaques after surgical removal of the uterus2. In fact, selective transmission of HIV through the vaginal mucosa rather than the cervix may commonly occur, as suggested by a recent large, randomized, controlled, prevention clinical trial in African women. In this study, no significant reduction in HIV‑1 acquisition occurred in women using a diaphragm compared with the con‑trol group3. However, the observed potential benefit of blocking HIV‑1 exposure to the cervix may have been undermined because the sexual partners of the women in the group using a diaphragm reported lower condom use than those in the control group.The region where the ectocervix transforms into the endocervix (FIG. 1) can have enriched CD4+ T‑cell populations and therefore may be a particularly sus‑ceptible site for HIV entry. Whether HIV can cross the endocervical mucus plug, reach the uterine cavity and*Vaccine and Infectious Disease Institute,Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA. Departments of ‡Medicine, §Obstetrics and Gynecology, ||Global Health and Laboratory Medicine, University of Washington School of Medicine, Seattle, Washington 98195, USA. Correspondence to M.J.M., e‑mail: jmcelrat@ doi:10.1038/nri2302 Published online 12 May 2008Setting the stage: host invasion by HIV Florian Hladik*‡§ and M. Juliana McElrath*‡||Abstract | For more than two decades, HIV has infected millions of people worldwide each year through mucosal transmission. Our knowledge of how HIV secures a foothold at both the molecular and cellular levels has been expanded by recent investigations that have applied new technologies and used improved techniques to isolate ex vivo human tissue and generate in vitro cellular models, as well as more relevant in vivo animal challenge systems. Here, we review the current concepts of the immediate events that follow viral exposure at genital mucosal sites where most documented transmissions occur. Furthermore, we discuss the gaps in our knowledge that are relevant to future studies, which will shape strategies for effective HIV prevention.NATURe ReVIeWS |immunology ADVANCe ONlINe pUblICATION | |©2008Nature Publishing GroupNature Reviews | Immunologyab Simian–HIV (SHIV). SHIVs are chimericviruses that are created byinserting the envelope protein(Env), the transcriptionaltransactivator (T at) and the regulator of virion geneexpression (Rev) of HIV into theSIV MAC239 clone. Depending onthe particular HIV Env protein,these SHIVs have different in vivo characteristics. The SHIV chimeric viruses are bestused for testing antibodiesspecific for HIV in non-humanprimate models. invade through the mucosa of the upper genital tract has not been well examined. In principle, uterine tissue is susceptible to infection if directly inoculated with HIV 4. Uterine simian–HIV (SHIV) infection has been shown in one monkey following vaginal inoculation two days earlier, an interval that is probably too short for stromal or lymphatic spread from the lower genital tract 5. This indicated that ascent of the virus through the endocervical mucus plug may be possible, but this observation has not been confirmed in humans. Conceivably, the upper mucosal tract may become more vulnerable to HIV‑1 penetration during ovula‑tion, a period when rising oestrogen levels alter the endocervical mucus making its consistency less viscous and more alkaline.both(FIG. 2)6–10. This has been shown in vivo in female macaques 11 and in mice 12, and13. studies using human cervical explants have also (REFS 14,15). Initially, cervical mucus can trap16,17. Conceivably, HIV virions that are initially free, or those that are transcytosis , endocytosis and(FIG. 2). The understanding of these events has been 18–20. Transcytosis tissue. On release from the epithelial cells, the virions readily infect susceptible leukocytes 21,22. Interestingly,cell‑associated virions secreted from infected seminal leukocytes appear markedly more efficient at transcy‑tosis than cell‑free virions 7,10,21. It has been reported that virions can also productively infect the cervical epithelial cells themselves 6, although this remains controversial 18,23. Conceivably, HIV‑1 can also be transported through the cervicovaginal epithelium to the draining lymphatics by donor lymphocytes and macrophages, as has been suggested in mouse studies 6,24. Our ex vivo experiments using sheets of isolated vagi‑nal epithelium, devoid of mucosal stroma, confirmed that HIV‑1 virions are sequestered in endocytic com‑partments and in the cytosol of epithelial cells (FIG. 3).Interestingly, although the experimental conditions permitted HIV‑1 access to both the luminal and basalsides of the epithelium, the virions were detected exclu‑sively in the basal and suprabasal epithelial cells (F.H., p . Sakchalathorn and M.J.M., unpublished observations). This suggests that initially, rather than entering and traversing superficial epithelial cells in the vagina and ectocervix, HIV‑1 probably disperses through the nar‑row gaps between them 16, as depicted in FIG. 2. This route might then permit HIV‑1 to directly contact and infect intraepithelial Langerhans cells (lCs) and CD4+ T cells 25 (see later), or it might allow HIV‑1 to reach suprabasal or basal epithelial cells that are more susceptible to viral sequestration and transcytosis. Importantly, components of human semen, most notably amyloid fibrils that form the single-layer columnar epithelium of the endocervix. The endocervical canal is filledwith mucus, providing a barrier against the ascent of pathogens. However, ovulation isaccompanied by hydration and alkalinization of the mucus plug, possibly decreasing itsbarrier function. Infection in women can also ensue when HIV-1 invades the single-layer columnar epithelium of the rectum following receptive anal intercourse. b | Inmen, viral invasion occurs most frequently through the inner foreskin and the penileurethra as a consequence of penile–vaginal or penile–anal intercourse. Thinly stratified columnar epithelial cells line most of the urethra except for the fossa navicularis near the external meatus (exit hole), which is covered by non-keratinized squamous epithelium. The glans penis and the outer foreskin are protected by keratinized squamous epithelium, which provides a strong mechanical barrier against HIV invasion. By contrast, a thin and poorly keratinized squamous epithelium covers the innerforeskin, rendering this site vulnerable to HIV invasion. Men are also infected by viralinvasion through the rectum.| ADVANCe ONlINe pUblICATION/reviews/immunol© 2008Nature Publishing GroupT ranscytosisThe process of transport of material, including HIV virions, across a cell layer by uptake on one side of the cell into a coated vesicle. The vesicle might then be sorted through the trans -Golgi network and transported to the opposite side of the cell, where itscontents are released into the extracelluar space.Langerhans cell(LC). A type of dendritic cell that is localized in thesquamous epithelial layer of the skin and certain mucosae.SyndecansSingle transmembrane domain proteins that carry three to five heparan sulphate andchondroitin sulphate chains that allow for interaction with various ligands including residues on the HIV-1 gp120 protein.from naturally occurring fragments of seminal prostatic acidic phosphatase, can capture virions and promote their attachment to epithelial cells and leukocytes, thus increasing infectivity 26.Several proteins expressed on the surface of epithelial cells may mediate the attachment of HIV‑1. Two cell‑surface glycosphingolipids, sulphated lactosylceramide expressed by vaginal epithelial cells 27 and galactosyl‑ceramide expressed by ectocervical epithelial cells 18,28, bind HIV‑1 gp120 and foster transcytosis 21. Interactions of HIV‑1 gp120 with transmembrane heparan sulphate proteoglycans (syndecans ) expressed by genital epithe‑lial cells can also contribute to HIV‑1 attachment and entry 19,22. Recently, gp340, a splice variant of salivary agglutinin that is expressed by cervical and vaginal epithelial cells, was shown to specifically bind to the HIV envelope protein and to enhance the passage of HIV through the epithelium giving it access to suscep‑tible leukocytes 29. One research group found that the β1 subunit of integrins expressed by cervical epithelial cells from some explants, but not all, bound virions that were presumably coated with fibronectin, which is abundant in human semen 16. Detection of HIV‑1 chemokine co‑receptor expression has been inconsist‑ent: one study did not detect the expression of either CC‑chemokine receptor 5 (CCR5) or CXC‑chemokine receptor 4 (CXCR4) by cervical epithelial cells 18, another reported the expression of CXCR4 by these cells 20, whereas another reported the exclusive expression of CCR5 (REF . 28).Regardless of the mode, the penetration of virus through the cervicovaginal epithelium in vivo occurs rapidly within 30–60 minutes of exposure, as shown in SIV‑infected macaques 30. Once within the epithelium,Table 1 |Contribution of HIV invasion sites to global HIV infections* *Table adapted from the UNAIDS/WHO AIDS epidemic update and men. §Includes MSM, bisexual men and women infected via anal receptive intercourse. ||Mother-to-child transmission. ¶Mostly intravenous drug use, butincludes infections by transfusions and health-care-related accidents. GI, gastrointestinal.NATURe ReVIeWS | immunologyADVANCe ONlINe pUblICATION |© 2008Nature Publishing GroupActivated memory T cell Resting memory T cellLangerhans cellEpithelial cell Stromal dendritic cell Array(DCs), reach close to the luminal surface of the mucosa. Monocytic precursor cells differentiate on arrival either intomacrophages or DCs, and DCs may differentiate further into subsets. Three stromal DC subsets have been identified in humanskin, distinguished by blood DC antigen 1 (BDCA1), CD1 and CD14 expression patterns64, but their presence and susceptibilityto HIV have not been determined in the mucosa. Infected donor cells and free virions may migrate along the abrasion anddirectly contact various target cells in the mucosal epithelium and stroma. Resident mucosal leukocytes such as DCs andT cells tend to cluster in these regions, creating susceptible foci for infection. Characteristic phenotypic cell receptors andreceptors relevant for HIV binding and infection are shown on the top of the figure (A). The possible pathways of HIVpenetration are summarized in B. a | Free HIV virions or HIV-infected donor cells are trapped in mucus, resulting in penetrationof the free virions into gaps between epithelial cells or attachment of HIV-infected donor cells to the luminal surface of themucosa and secretion of virions on contact. The virions are then captured and internalized into endocytic compartments byLangerhans cells that reside within the epithelium. b | HIV can also fuse with the surface of intraepithelial CD4+ T cells, followedby productive infection of these cells. c | Infected donor cells or free virions can immigrate along physical abrasions of theepithelium into the mucosal stroma. There they are taken up by lymphatic or venous microvessels and transported to locallymph nodes or into the blood circulation, respectively, or they make contact with stromal DCs, T cells and macrophages.d | Virions can transcytose through epithelial cells near or within the basal layer of the squamous epithelium (see also Figure 3),productively infect basal epithelial cells, be internalized into endocytic compartments, or penetrate between epithelial cells.e | Once within the stroma, virions can productively infect stromal DCs or be internalized into the endocytic compartments ofDCs and pass from the stromal DCs to CD4+ T cells across an infectious synapse (see also Figure 4) where massive productiveinfection of CD4+ T cells ensues. In addition, virions can productively infect resting mucosal CD4+ memory T cells in the stromaand possibly stromal macrophages. f | Productively infected CD4+ T cells and stromal DCs, and stromal DCs or intraepithelialLCs harboring virions in endocytic compartments, can emigrate into the submucosa and the draining lymphatic and venousmicrovessels. CCR5, CC-chemokine receptor 5; DC-SIGN, dendritic-cell-specific ICAM3-grabbing non-integrin.| ADVANCe ONlINe pUblICATION /reviews/immunol©2008Nature Publishing GroupNature Reviews | Immunology DesmosomesNucleusBasal cell layera200nm2µmStromal papillaeSuperficial areas of themucosal stroma that interdigitate with the epithelium.C-type lectin receptorsA large family of receptors that bind glycosylated ligands and have multiple roles, such as in cell adhesion, endocytosis, natural-killer-cell targetrecognition and dendritic-cell activation.R5-tropic HIV-1An HIV strain that uses CC-chemokine receptor 5 (CCR5) as the co-receptor to gain entry to target cells.HIV encounters CD4+ T cells as well as lCs. lCs have dendrites that extend and retract through the intercel‑lular spaces 31, and which can even reach up to the surface of the epithelium 32, where HIV can bind directly to them (T. Hope, personal communication). based on observa‑tions of gut DCs 33,34, this could also be particularly true for DCs that are located just beneath the endocervical columnar epithelium. However, direct sampling of luminal pathogens by endocervical DCs or vaginal lCs, which could be exploited by HIV to bypass the epithelial‑cell barrier, has not yet been formally demonstrated. Mechanical micro‑abrasions of the mucosal surface induced by intercourse may also allow HIV to directly access target cells, such as DCs, T cells and macrophages, at the basal epithelium and the underlying stroma 35. Areas above the stromal papillae , where the epithelium is relatively thin and where lCs on the epithelial‑cell side (F.H., l. ballweber and M.J.M., unpublished observa‑tions) and T cells and macrophages on the stromal side 28 congregate, appear particularly vulnerable to viral inva‑sion. Consistent with this notion, in vivo simian immu‑nodeficiency virus (SIV) infection of the genital mucosa of macaques is initially established in a highly focal man‑ner, and continuous seeding from this nidus of infection is crucial for establishing systemic infection 17. Similarly, chemical micro‑abrasions from the use of certain topical microbicides and micro‑abrasions due to genital ulcers caused by sexually transmitted diseases (for example, syphilis, chancroid and those caused by herpes simplex virus) are also likely to result in the exposure of vulnerable target cells in the basal epithelium and stroma 36.Importance of cervicovaginal LCs in HIV invasion. For many years, HIV‑1 invasion of the lower genital tract has been assumed to occur through the internalization of HIV‑1 by lCs. This view was supported by evidence that skin lCs are susceptible to infection by HIV‑1 (REFS 37–40) and that genital mucosal lCs harbour SIV virions within 24 hours of intravaginal inoculation of macaques 30. However, soon after ex vivo organ culture, lCs migrate out of the epithelium 14,23,41–43; therefore, examination of lC infection specifically within the human vaginal epithelium has been technically difficult. For example, one landmark study demonstrated that after exposure of complete cervical mucosa from humans to HIV‑1, emigrating DCs had efficiently captured HIV and were capable of transmitting the virus in trans 42. However, it was not possible to determine whether the cells originated from the epithelium as lCs or from the underlying stroma as DCs. More recently, we resolved this issue by preparing sheets of vaginal epithelium separated from the underlying stroma, and observed that vaginal lCs efficiently internalized HIV‑1 into their cytoplasmic compartments 25. As lCs exit the epithelium at the basal side, they transport intact virions, thereby enabling the infection to spread beyond the site of viral entry (FIG. 2). However, it is not clear if lCs in the cervicovaginal epithelium can produce and release new HIV‑1 virions. HIV receptors expressed by these lCs include CD4, CCR5 and the C-type lectin receptor langerin (also known as CD207), but not CXCR4 and DC‑specific ICAM3 (inter‑cellular adhesion molecule 3)‑grabbing non‑integrin (DC‑SIGN; also known as CD209)25,44–47. Antibodies that bind CD4 and CCR5 partially block the uptake of R5-tropic HIV-1 by lCs, but blocking the binding of HIV‑1 to C‑type lectin receptor with mannan, a mannose polymer, had little effect on uptake 25. Although low‑level CD4‑ and CCR5‑mediated productive infection of lCs by HIV‑1 in human skin explants has been shown 39,40,48, we were unable to confirm this finding in our imaging studies of vaginal lCs 25. Therefore, if de novo production of virions occurs, it appears relatively inefficient in con‑trast to the high capacity of vaginal lCs to endocytose HIV‑1. Nevertheless, even low levels of productive HIV‑1 infection of cutaneous DCs and lCs lead to profound viral replication in co‑cultured T cells 40,48,49. Therefore, future investigations must conclusively determine if lCs in cervicovaginal epithelium can support productive HIV infection in vivo and if this property is required for the passage of the virus to T cells, as has been reported for other types of DC 50–53.The relative inefficiency of mannan in blocking the binding and endocytosis of HIV‑1 by vaginal lCs was sur‑prising 25 because C‑type lectin receptors, which recognize mannose‑containing carbohydrate structures, mediate viral entry in other types of DCs 46. However, HIV‑1 can bind DC subsets independent of C‑type lectin receptors and CCR5 (REFS 54–56). So, although HIV virions can efficiently bind to langerin expressed by epidermal lCs for their entry into these cells 57, they appear to largely bypass binding to langerin expressed by vaginal lCs in favour of alternative endocytic routes. This distinction may be highly relevant for mucosal HIV transmission.Figure 3 | HiV-1 transcytosis in situ in the vaginal epithelium. Electron micrographshowing a vaginal epithelial cell located one layer above the basal cell layer andcontaining intact cytoplasmic virions following 24 hours of infection with HIV-1BaL (a procedure performed on vaginal epithelial sheets, as previously described 25) (a ). As shown in the higher power insets, the virions can be seen on both the apical (b ) and basal (c ) sides of the nucleus, signifying transcytosis. Desmosomes and keratin fibres,distinguishing features of the epithelial cell, can also be seen (c ).NATURe ReVIeWS | immunologyADVANCe ONlINe pUblICATION |© 2008Nature Publishing GroupBirbeck granules Membrane-bound rod- or tennis-racket-shaped structures with a central linear density, found in the cytoplasm of Langerhans cells. Their formation is induced by langerin, an endocytic C-type lectin receptor that is specific to Langerhans cells. PhagosomesVacuolar compartments that confine microorganisms after enforced endocytosis orafter phagocytosis. Unless counteracted by a microbial survival strategy, the phagosome matures into a hostile environment that is designed to kill and digest microorganisms.Cross-presentationThe initiation of a CD8+ T-cell response to an antigen that is not present within antigen-presenting cells (APCs). This exogenous antigen must be taken up by APCs and then re-routed to the MHC-class-I pathway of antigen presentation.Lamina propria Connective tissue that underlies the epithelium of the mucosa and contains various myeloid and lymphoid cells, including macrophages, dendritic cells, T cells andB cells. langerin expressed by epidermal lCs can direct HIV‑1to Birbeck granules for degradation57. by contrast, by gain‑ing entry to the vaginal lCs independently of langerin,HIV may survive by reaching endocytic compartments,such as early phagosomes, where antigens are preservedfor cross-presentation58. This is consistent with our observa‑tions that intact virions were still present in lCs that hadmigrated out of the vaginal epithelium 60 hours after viralchallenge25. Therefore, it appears that HIV‑1 enters vaginallCs through a different route than when it enters skin lCs,and this results in a distinct fate of the endocytosed viri‑ons. More detailed studies are now warranted to uncoverwhich endocytic pathways HIV uses in vaginal lCs, andhow this can be harnessed therapeutically.Infection of DCs in the cervicovaginal stroma. Unlikegenital lCs, stromal DCs express both DC‑SIGN46,47and CCR5 (REFS 59,60) and have been implicated inSIV and HIV infection, but their exact role in mucosaltransmission is not clear. In situ studies in the humanexplant model have failed to identify infected DCs in thecervicovaginal stroma14,23,41,43. by contrast, SIV‑infectedDCs were present in the lamina propria of the cervico‑vaginal mucosa of macaques shortly after intravaginalSIV challenge30,61, as well as in chronically infected ani‑mals62. likewise, HIV‑infected DCs were identified intissue biopsies of the vaginal stroma of asymptomaticHIV‑1‑infected women63.The failure to identify infection of stromal DCs in thehuman explant models might be attributable to the rela‑tively low sensitivity of the detection methods used andthe migration of stromal DCs from the tissue, which maydrastically decrease the number of infected cells in situover time. Indeed, when DCs were harvested from theculture supernatants of human cervical explants that werechallenged with HIV‑1, significant in trans infectivity wasdetected42 and massive budding of virions was observedamong emigrant DCs 5 days after virus exposure60(FIG. 4).However, it could not be determined if the original sourceof DCs was from the epithelium proper or from the stro‑mal tissue. In addition, inferring the initial susceptibilityof DCs while confined to the mucosal stroma, based onfindings from emigrated DCs that undergo phenotypicchanges as they exit the mucosa, may be less reliable.Therefore, stromal DCs exhibit different HIV‑1 recep‑tor expression patterns compared with lCs, potentiallypermitting different HIV‑1 entry pathways. Much stillremains to be learned about stromal DCs in the humangenital mucosa in general, whether different subsets existsimilar to those found in skin dermis64, and the interactionof these DC subsets with HIV virions in particular. Moresensitive in situ detection methods of HIV infection, aswell as assays that can distinguish de novo virus produc‑tion from endocytically engulfed virions, as recentlyreported65, will be helpful in determining the exactcontribution of stromal DCs to the propagation of HIV.HIV infection of cervicovaginal CD4+ T cells. CD4+T cells are dispersed throughout the lamina propria ofthe human vagina, ectocervix and endocervix, oftenclustering just beneath the basal membrane66,67. They arealso found at variable numbers within the vaginal andectocervical squamous epithelium66,67. The majority ofthese cells are memory T cells that express higher levelsof CCR5 than those that circulate in the blood25,68–70. Oneday after HIV‑1 inoculation of vaginal, ectocervical andendocervical tissue cultures, infected CD4+ T cells wereshown to be confined to the mucosal stroma14,16,23,42.This result was surprising, given the presence of CCR5+CD4+ T cells within the squamous epithelium. However,by analysing the fate of fluorescently tagged virions asearly as 2 hours after viral exposure, we observed thatR5‑tropic HIV‑1 efficiently bound to intraepithelialvaginal CD4+ T cells, and this was followed by fusionand productive infection25. Therefore, infected T cellsmust rapidly leave the epithelium, and those found inthe stroma may be the same or the early progeny ofintraepithelial T cells.Findings in the human explant studies show thatHIV‑1 very effectively targets CD4+ T cells in thegenital mucosa for productive infection14,25,42, and thatthe initial infection of intraepithelial CD4+ T cells isprobably independent of lCs25. The central role forgenital CD4+ T cells in early infection and propaga‑tion is also evident from SIV challenge experimentsin macaques62,71,72. Interestingly, not only does SIVproductively infect activated T cells, characterized byHlA‑DR and Ki67 expression, it also infects T cells thatare in the resting state (HlA‑DR– and Ki67– T cells)71.Consistent with this finding, we observed binding ofHIV‑1 to both HlA‑DR+ and HlA‑DR– intraepithelialT cells in our human vaginal explant model (M.J.M.,p. Sakchalathorn, l. ballweber and F.H., unpublishedobservations). In addition, the contribution of restingCD4+ T cells to viral production is substantial duringthe very earliest stages of infection73. The fact that vagi‑nal CD4+ T cells are rapidly depleted following intra‑venous SIV inoculation of macaques5,72, similar to thedepletion of CD4+ T cells in the gut during acute SIVinfection74, further illustrates their high susceptibilityto infection in vivo.Other leukocyte HIV targets in the female genital tract.Macrophages in the female genital mucosa are alsosusceptible targets for early HIV‑1 infection, as demon‑strated in studies using human cervical explant mod‑els23,41,43, and in two reports these cells were the major celltype that was infected by R5‑tropic HIV‑1 (REFS 23,43).Whether or not resident macrophages in the femalegenital tract constitutively express CCR5 in situ is notknown, but most macrophages do so when harvestedfrom supernatants of vaginal organ cultures60, suggestingthat the expression of CCR5 by macrophages may occurduring the period of activation and emigration fromthe mucosa75. by contrast, SIV‑infected macrophagesin genital tissues were either rare71, or undetectable30,61.likewise, macrophages in the human intestinal mucosawere reported to lack CCR5 expression and to possesslow permissibility to HIV‑1 infection76. These discrep‑ancies illustrate a potential limitation of organ cultures.If explantation activates stromal macrophages and as aconsequence increases surface CCR5 expression, this| ADVANCe ONlINe pUblICATION /reviews/immunol©2008Nature Publishing Group。
stochastic calculus for fractional brownian motion and related processes附录
kH (t, u)dWu = CH Γ (1 + α)
(2)
R
α (I− 1(0,t) )(x)dWx
(see Lemma 1.1.3). Therefore, the first equality is evident, since
0 R t
(kH (t, u))2 x)α )2 dx +
k n
2H
2
.
C . n2
(B.0.12)
References
[AOPU00] Aase, K., Øksendal, B., Privault, N., Ubøe, J.: White noise generalization of the Clark-Haussmann-Ocone theorem with applications to mathematical finance. Finance Stoch., 4, 465–496 (2000) [AS96] Abry, P., Sellan, F.: The wavelet-based synthesis for fractional Brownian motion proposed by F. Sellan and Y. Meyer: Remarks and fast implementation. Appl. Comp. Harmon. Analysis, 3, 377–383 (1996) [AS95] Adler, R.J.; Samorodnitsky, G.: Super fractional Brownian motion, fractional super Brownian motion and related self-similar (super) processes. Ann. Prob., 23, 743–766 (1995) [ALN01] Al` os, E., Le´ on, I.A., Nualart, D.: Stratonovich stochastic calculus with respect to fractional Brownian motion with Hurst parameter less than 1/2. Taiwanesse J. Math., 5, 609–632 (2001) [AMN00] Al` os, E., Mazet, O., Nualart, D.: Stochastic calculus with respect to fractional Brownian motion with Hurst parameter less than 1/2. Stoch. Proc. Appl., 86, 121–139 (2000) [AMN01] Al` os, E., Mazet, O., Nualart, D.: Stochastic calculus with respect to Gaussian processes. Ann. Prob., 29, 766–801 (2001) [AN02] Al` os, E., Nualart, D.: Stochastic integration with respect to the fractional Brownian motion. Stoch. Stoch. Rep., 75, 129–152 (2002) [And05] Androshchuk, T.: The approximation of stochastic integral w.r.t. fBm by the integrals w.r.t. absolutely continuous processes. Prob. Theory Math. Stat., 73, 11–20 (2005) [AM06] Androshchuk, T., Mishura Y.: Mixed Brownian–fractional Brownian model: absence of arbitrage and related topics. Stochastics: Intern. J. Prob. Stoch. Proc., 78, 281–300 (2006) [AG03] Anh, V., Grecksch, W.: A fractional stochastic evolution equation driven by fractional Brownian motion. Monte Carlo Methods Appl. 9, 189–199 (2003)
1p36.33-32缺失所致婴儿痉挛症1例并文献复习
甘肃医药2021年40卷第6期Gansu Medical Journal ,2021,Vol.40,No.61p36.33-32缺失所致婴儿痉挛症1例并文献复习王三萍赵淑珍祁俏英罗璇甘肃省人民医院,甘肃兰州730000【摘要】通过对1例表现为横眉、双眼稍凹陷、智力语言运动明显低下、难治性癫痫、痉挛发作的患儿进行染色体微矩阵检测,发现1p36.33-1p36.32区域缺失,探讨1p36.33-32缺失的临床特征与遗传学特点。
应用促肾上腺皮质激素(ACTH )、多种抗癫痫药、生酮饮食治疗,治疗有效但易复发,产前诊断可能有效减少本病的发生。
【关键词】1p36缺失;婴儿痉挛症;染色体微阵列分析技术中图分类号:R742.1文献标识码:B文章编号:1004-2725(2021)6-0570-03·短篇及个案报告·1p36缺失是最常见的染色体亚端粒区域微缺失,患病率在活产新生儿中约为1/5000[1]。
主要表现为中度到重度智力障碍、听、视力障碍、肌张力低、脑畸形、癫痫、心脏畸形等[2]。
lp36缺失综合征发病率高,但由于表现多样化,所以临床医生对此征的认识鉴别仍有困难。
目前国内报道2例[3],表现生长发育迟缓,尚无婴儿痉挛症相关报道。
本文回顾分析经染色体微阵列分析技术基因检测确诊的1例患儿,并复习文献。
1临床资料患儿,男,8月龄,因反复抽搐3月就诊,抽搐早期表现为部分性发作,双眼一侧斜视,口周发绀,伴或不伴四肢抽搐,10余秒缓解,给予左乙拉西坦治疗,发作缓解。
1月后出现点头弯腰样成串发作,每天2~8次不等,生长发育迟缓,不能俯卧抬头、翻身、抓物,视觉追踪差。
个人史及家族史:足月顺产,G3P3,出生身长50cm ,体重2.5kg 。
母亲怀孕期间孕酮低,无流产史,四维超声提示侧脑室略增宽,9mm ,唐氏筛查及无创DNA 检测阴性,家族无遗传史。
查体:神清,身高正常,眼窝稍深,一字眉(面部特征见图1),前囟门3cm ×3cm ,喉软骨发育不良,心前区未闻及杂音,外生殖器未见异常。
绿色荧光蛋白及其应用
p-HBI 生色团的成熟过程经历 GFP 多肽骨架折叠 和生色团形成两个阶段,期间 4 个保守氨基酸残基发 挥着特殊的功能作用[10]。
2011,31( 1)
邓 超 等: 绿色荧光蛋白及其应用
97
图 1 野生型 avGFP 的结构 Fig. 1 The structure of wild type avGFP
4 不同类型的荧光蛋白
通过 定 点 突 变 和 随 机 突 变 得 到 了 不 同 突 变 型 的 avGFP 样蛋白,珊瑚类荧光蛋白的发现使人们发展出更 多性质各异的荧光蛋白,发射谱覆盖 420 ~ 655 nm,应 用范围不断扩大 [14-15]。部分荧光蛋白及基本性质见表 1 所示。 4. 1 蓝色荧光蛋白
2 绿色荧光蛋白的结构
从维 多 利 亚 多 管 水 母 中 分 离 出 来 的 野 生 型 GFP ( avGFP ) 由 238 个 氨 基 酸 残 基 组 成,分 子 质 量 约 27kDa,二级结构包括 11 个 β 折叠链( β-sheet strand) , 8 个螺旋( helix) ,3 个转折( turn) [图 1 ( a) ],三维结构 ( PDB 登录号 1EMA 和 1GFL,1GFL 为二聚体) 为 42 × 24 ( 高 × 直径) 的 β 圆柱( β-barrel) ,圆柱两端由一 些较短的 α 螺旋盖住,圆柱中央是几段 α 螺旋,生色团 的三肽( Ser65-Tyr66-Gly67) 位于圆柱中央[图 1 ( b) ~ ( d) ]。该结构性质稳定,圆柱内部的微环境对维持生 色团的正确构象从而产生荧光以及保护生色团不被氧 气淬灭等都有重要作用[10][图 1( e) ]。
荧光蛋白的寡聚可能会影响融合蛋白的正确定位和迁移几乎所有荧光蛋白都有寡聚趋势通过对有相互作用的侧链氨基酸进行突变可消除这种趋如蓝色荧光蛋白激发光接近紫外光一些珊瑚类荧光蛋白细胞毒性已有报道荧光蛋白发展至今人们对其在研究生物大分子相互作用及时空变化中的重要作用已没有质疑但相对于复杂的生命来说荧光蛋白还不足以解决许多问题
临床基因组学检验:染色体微阵列技术原理 与临床应用
Genetic/Genomic Disorders
Genomic disorders (number) Trisomy 21 Trisomy 18 Trisomy 13 Mosaic trisomies of other chromosomes
Genomic disorders (structure) More than 400 known disorders
longevity
More than 150 genes prevents cholesterol buildup
精确控制胚胎发育和分化的每一个步骤; 决定了个体的所有生命特征; 决定了个体患各种疾病的可能性.
遗传病:遗传物质发生突变所引起的疾病。 种类:确定的遗传疾病超过7000种。 1、单基因病--涉及一对基因,AR、AD、XR、XD、Y连 锁遗传病。隐性遗传4000,显性遗传3000。 2、多基因病--多对基因和环境共同作用所导致的疾病。 3、染色体病--数目异常及结构异常引起的疾病。 4、体细胞遗传病--体细胞突变如肿瘤。 5、线粒体病--线粒体功能异常为主要起因的一大类疾病。 特征:垂直传递、终生性、发病率低、危害严重、家族性 发病、多无有效治疗。成为危害人类健康的主要疾病。
出生缺陷
出生缺陷
遗传因素
染色体异常(所有新生儿中,染色 体异常占0.92%,多为新发而非遗 传)
单基因突变(多为孟德尔遗传,少 数为新发)
环境因素(理、化、生物因素、生活方式)
遗传+环境因素
基因组:细胞核DNA成分和线粒体DNA分子的总和 基 因: 基因组内一个个具体的结构和功能单位 染色体:基因的载体
复,被収现广泛存在于人类基因组中[1,2]。已有大量研究证实CNVs
The largest outbreak of hand; foot and mouth disease in Singapore in 2008
The largest outbreak of hand;foot and mouth disease in Singapore in2008: The role of enterovirus71and coxsackievirus A strainsYan Wu a,b,Andrea Yeo b,M.C.Phoon a,E.L.Tan b,c,C.L.Poh d,S.H.Quak b,Vincent T.K.Chow a,*a Department of Microbiology,Yong Loo Lin School of Medicine,National University of Singapore,Kent Ridge117597,Singaporeb Department of Paediatrics,National University Hospital,Singaporec School of Chemical and Life Sciences,Singapore Polytechnic,Singapored Swinburne University of Technology,Victoria,Australia1.IntroductionHand,foot and mouth disease(HFMD)is a common childhooddisease characterized by a brief febrile illness,typical vesicularrashes on the palms,soles,or buttocks,and oropharyngeal ulcers.In rare cases,patients may also develop neurological complica-tions,such as encephalomyelitis,aseptic meningitis,and acuteflaccid paralysis.1,2The most common etiologic agents arecoxsackievirus A16(CA16)and enterovirus type71(EV71).However,throughout an outbreak,many other enteroviruses,such as coxsackieviruses A4and A6and certain echoviruses,mayco-circulate.EV71has been strongly associated with neurologicalcomplications,especially during large HFMD epidemics triggeredby EV71within the Southeast and East Asian regions in recenttimes.3In2008,the largest outbreak of HFMD in Singapore afflicted29686patients ranging from kindergarten to primary schoolstudents,four of whom developed EV71-related encephalitis.Moreover,a3-year-old boy with EV71infection died of encepha-lomyelitis in August2008–thefirst HFMD-related death since2001.4Although the HFMD cases surged to an unprecedented highlevel,only a single fatality was documented,suggesting that thestrains responsible for the2008Singapore outbreak caused highlytransmissible but relatively mild disease.In this study,clinical samples from HFMD patients wereinvestigated in order to identify the circulating virus serotypes inthe2008HFMD outbreak in Singapore.The predominantenteroviruses and EV71subgenogroups were characterized byphylogenetic and genome sequence analyses.2.Materials and methods2.1.Patients,clinical specimens,virus isolation,and statistical testsThis study was approved by the Institutional Review Board ofthe National University of Singapore(Approval No.NUS-301).Between March13and August15,2008,clinical samples wereobtained from43children presenting with suspected HFMD at theInternational Journal of Infectious Diseases14(2010)e1076–e1081A R T I C L E I N F OArticle history:Received8April2010Received in revised form14July2010Accepted20July2010Corresponding Editor:Jane Zuckerman,London,UK.Keywords:HandFoot and mouth diseaseHerpanginaLargest outbreakSingaporeEnterovirus71Coxsackievirus AHigh transmissibilityA B S T R A C TBackground:During2008,Singapore experienced its largest ever outbreak of hand,foot and mouthdisease(HFMD),resulting in29686cases,including four cases of encephalitis and one fatality.Methods:A total of51clinical specimens from43patients with suspected HFMD at the NationalUniversity Hospital,Singapore were collected for virus isolation and identification by reversetranscription polymerase chain reaction(RT-PCR)and sequencing.Results:Enteroviruses were identified in34samples(66.7%),with11samples(21.6%)being positive forenterovirus71(EV71).Other non-EV71enteroviruses(including coxsackievirus A4,A6,A10,and A16)were identified in23samples(45.1%).The most prevalent virus serotypes were CA6,CA10,and EV71.CA6and CA10accounted for35.3%of all HFMD cases,which may explain the high transmissibility andlow fatality that characterized this unprecedented epidemic associated with relatively mild disease.Phylogenetic analyses of10circulating EV71strains indicated that they belonged to two subgenogroups,i.e.,B5(80%)and C2(20%).The VP1sequences of the2008EV71strains also exhibited continuousmutations during the outbreak,reflecting the relatively high mutation rate of the EV71capsid protein,which may have implications for future vaccine development.Conclusions:A safe and effective vaccine against EV71is certainly warranted in view of its potentialneurovirulence and its role in HFMD epidemics of recurring frequency with resultant fatalities in Asia,aswell as other parts of the world.ß2010International Society for Infectious Diseases.Published by Elsevier Ltd.All rights reserved.*Corresponding author.Tel.:+6565163691;fax:+6567766872.E-mail address:micctk@.sg(Vincent T.K.Chow).Contents lists available at ScienceDirectInternational Journal of Infectious Diseasesj o u r n a l h o m e p a g e:w w w.e l se v i e r.co m/l oc a t e/i j i d1201-9712/$36.00–see front matterß2010International Society for Infectious Diseases.Published by Elsevier Ltd.All rights reserved.doi:10.1016/j.ijid.2010.07.006National University Hospital,Singapore.The study inclusion criteria were the presence of papules and/or ulcers on the palms, soles,or buttocks or in the mouth.A similar HFMD case definition was used in a previous study of the HFMD outbreak in2000.4A total of51samples,including throat swabs,nasal swabs,rectal swabs,saliva,urine,and blood were collected.Throat,nasal,and rectal swabs were transferred into virus transport medium.For virus isolation,all specimens were immediately inoculated upon receipt into the human rhabdomyosarcoma(RD)cell line,and cultured for up to three passages.5,6Pathogens other than enteroviruses were not investigated.The statistical differences between proportions were tested using the Chi-square test or Fisher’s exact test.2.2.RNA extraction and reverse transcriptionViral RNA was extracted using the QIAamp viral RNA mini kit (Qiagen,Hilden,Germany)according to the manufacturer’s instructions.Reverse transcription(RT)was performed to synthe-size cDNA using MMLV(Moloney murine leukemia virus)reverse transcriptase.72.3.EV71real-time RT-PCR hybridization assayEach specimen was subjected to a real-time one-step RT-PCR hybridization assay with EV71-specific primers and probes FL and LC using the LightCycler RNA amplification hybridization probe kit (Roche Applied Science,Indianapolis,IN,USA),as reported previously.72.4.Classical RT-PCR and direct sequencing for enterovirusidentificationSeveral pairs of primers were employed for the identification of enteroviruses.Firstly,pan-enterovirus(pan-EV)primers were used for enterovirus detection by amplifying a target within the50 untranslated region(50UTR).8To confirm the enterovirus type, 50UTR-F and50UTR-R primers were employed to amplify a segment also within the50UTR of all enteroviruses,9and amplified products were subjected to direct DNA sequencing.For EV71detection,VP3-Fa and EV2A-R primers targeting the VP1gene of all EV71genogroups (50-ATWWTRGCAYTRGCGGCRGCC-30and50-TCGCKRGAGCTGT CTTCCCAVA-30,respectively)were newly designed to amplify a 1200-bp segment,followed by DNA sequencing.To confirm the identity of non-EV71enteroviruses,primers were designed to amplify the30segment of VP110for CA4(50-CCTAAGCCTGATGCCC-GAGA-30and50-TTGTGATCTCAAAGGCCTAGGGA-30),CA6(50-GTGTCCGTCCCATTCATGTC-30and50-GTTCTCTGTGGGTCTGCTGG-30),CA10(50-AAACCGACTGGAAGGGATGC-30and50-CGATCTCGTG-CACTGTTGGC-30),and CA16(50-TGAAAATGACGGACCCACCA-30and 50-ATCTTGTCTCTACTAGTGCTGGTG-30).2.5.DNA sequencing,nucleotide and amino acid sequences,and phylogenetic analysesAll amplicons were sequenced in both directions using the BigDye cycle sequencing kit and ABI automated DNA sequence analyzer(Applied Biosystems,Carlsbad,CA,USA).Sequences were subjected to BLAST analysis(/blast),while ClustalW(/tools/clustalW2/index.html)was ap-plied for multiple sequence alignment.11A phylogenetic tree was constructed based on EV71VP1nucleotide sequences of10 Singapore2008strains together with selected strains isolated from different geographic regions from1974to2003that were used to construct the tripartite genogroup structure of EV71.12In addition, two strains from fatal cases of the2000Singapore outbreak and 2008China outbreak were selected for comparative sequence analysis.The dendrogram was constructed by the neighbor-joining method using MEGA 4.0software,and bootstrapping was performed from1000replicates.3.Results3.1.Clinical features of patients with EV71versus non-EV71infectionsClinical data were available for42out of43patients;most of them(n=37,88.1%)received outpatient treatment at the Children’s Emergency Department,while only12.0%(n=5)were hospitalized on the pediatric ward.The number of females and males was similar,and the majority of patients(n=28,66.7%)were under5years of age.With regard to age group,the highest percentages of patients were children aged3years(40%of EV71 and19%of non-EV71)and4years(20%of EV71and22%of non-EV71).Figure1compares the symptoms of HFMD patients infected with EV71and other enteroviruses.None of the patients presented with severe symptoms,and there were no fatal or encephalitis cases.Notably,papules were observed for all cases,while mouth ulcers were also seen in almost all cases,i.e.,90%(n=10)for EV71 versus100%(n=32)for non-EV71infections.Papules and mouth ulcers are two of the clinical criteria for the diagnosis of HFMD and herpangina.Interestingly,more thanfive papular lesions per patient were noticed in75%(n=9)of those positive for EV71and CA16,compared to only43%(n=13)of other patients,but this was not statistically significant(p>0.05).Rhinorrhea was only observed in31%(n=10)of non-EV71infections and not in any EV71infections.Poor feeding was documented in30%(n=3)of EV71cases and in25%(n=8)of non-EV71infections.With regard to other clinical signs,non-EV71patients displayed these more frequently,but the differences were not statistically significant (p>0.05).For example,the difference was about two-fold for cough and vomiting,while diarrhea was only occasionally observed.3.2.Pan-enterovirus RT-PCR,direct sequencing,and virus isolation elucidate the distribution of enterovirus types and the involvement of EV71in HFMD patientsOnly one clinical sample was collected from each of37patients, while three different clinical samples were collected from two patients and two from four patients,giving a total of51samples from43patients.Table1summarizes the detection rate based on clinical sample type,and highlights the throat swab as a good clinical specimen for HFMD virus detection and isolation.ThisFigure1.Distribution of clinical features in EV71and non-EV71patients.Y.Wu et al./International Journal of Infectious Diseases14(2010)e1076–e1081e1077finding is not surprising since substantial amounts of virus are expected in patients with multiple vesicles and/or ulcers in the oral mucosa.We did not collect vesicular fluid samples from our patients as their parents perceived this procedure to be invasive and painful.Classical RT-PCR assays with both pairs of pan-EV primers targeting the 50UTR were able to detect enteroviruses in 34samples (66.7%),with 17samples (33.3%)being enterovirus-negative (Table 2).RT-PCR with EV71-specific primers VP3-Fa and EV2A-R detected 11EV71-positive samples (21.6%).In addition,the non-EV71enterovirus types were identified via sequencing of 50UTR and VP1amplicons.The circulating enteroviruses responsi-ble for the 2008Singapore outbreak included CA4,CA6,CA10,CA16,and EV71,the most prevalent being CA6(23.5%),EV71(21.6%),and CA10(11.8%).It is noteworthy that CA4and CA16accounted for only 9.8%,even though CA16has played a major role in previous HFMD outbreaks.If the enterovirus-negative samples were excluded,more than 50%of the samples were positive for CA6and CA10,while 32%were EV71-positive,thus reiterating the predominant role of CA6,CA10,and EV71in this outbreak.This was corroborated by cell culture inoculation,which successfully isolated enteroviruses from 19out of 51samples (approximately 40%),later confirmed as nine CA6(47.4%),six CA10(31.6%),three EV71(15.8%),and one CA4(5.3%).The EV71real-time RT-PCR hybridization assay failed to detect EV71in all 51specimens.VP1sequence alignment of the 10EV71strains with the RT-PCR primers and hybridization probes revealed one mismatch in each primer.However,three mismatches were found for the probe with acceptor fluorophore spanning nucleo-tides 2519–2496,one of which was near the 30end (Figure 2)and may compromise binding of the probe to the target product leading to failure of detection.This mismatch of the latter probe rather than primers was supported by classical RT-PCR using the primers which could amplify specific bands for all EV71strains as detected by gel electrophoresis (data not shown).3.3.Molecular epidemiology of EV71outbreak strains identifies two major subgenogroupsOut of the 11EV71-positive samples,2were from the same patient,with the rest from individual patients.Thus,the distribution of EV71subgenogroups of 10strains was determined by RT-PCR amplification of their complete VP1fragments,followed by nucleotide sequencing and phylogenetic analyses.The sub-lineage structure of EV71was reconstructed,12and revealed two circulating subgenogroups (Figure 3)belonging to B5(eight strains or 80%)and C2(two strains or 20%).Three EV71strains were successfully isolated from the samples,two of which were B5and the other was C2.Interestingly,the VP1sequences of strains from the dominant B5subgenogroup displayed differences,whereas those of the two subgenogroup C2strains were identical.The complete viral genomes of the two representative subgenogroups B5(NUH0083/SIN/08)and C2(NUH0075/SIN/08)were sequenced and deposited in the GenBank database under accession numbers FJ461781and EU868611,respectively.The GenBank accession numbers for the other sequences are FJ461782–FJ461789and GU198753–GU198764.3.4.VP1sequence comparison reveals interesting disparities between 2008outbreak and known virulent strainsTwelve VP1gene sequence disparities of B5strains were identified,suggesting the occurrence of viral evolution and mutation during this outbreak (Figure 4).An interesting trend was noticed at nucleotides 19,373,and 756,whereby three B5samples obtained in April and May 2008(NUH0049,0047,0086)were identical at these positions,in contrast to the existence of disparities for the other five B5strains that were received later,during mid-May to August 2008(NUH0083,0085,0043,0037,0012).Furthermore,two B5strains that were collected later harbored non-conservative VP1amino acid substitutions,i.e.,K to E at position 215of NUH0043,and T to A at position 289of NUH0037.These phenomena provide strong evidence for virus mutational events through the course of the outbreak that may partly arise from strong immunological pressure on the immuno-dominant VP1region.In addition,for the 2008strains,disparities were discovered at three VP1epitopes that are capable of eliciting neutralizing antibodies against EV71in vitro and in vivo.13–15Table 1Identification of enteroviruses by classical and real-time RT-PCR and virus isolation from different clinical specimens.Virus detection techniqueThroat swab (n =38)Saliva (n =3)Nasal swab (n =1)Rectal swab (n =3)Urine (n =3)Foot ulcer (n =1)Blood (n =2)Overall positivity rate (%)Pan-EV 154bp 2630221066.7Pan-EV 439bp2630221066.7VP3-Fa and EV2A-R 820010021.6Real-time RT-PCR 00000000Virus isolation17237.2RT-PCR,reverse transcription polymerase chain reaction.Table 2Distribution of enteroviruses identified in clinical specimens.Enterovirus serotype Number of cases Percentage (%)CA43 5.9CA61223.5CA10611.8CA162 3.9EV711121.6EV-negative 1733.3Total5110025192496NUH0049/SIN/08–B5G C T G G C A G G G C C C G G G T G A G C G C C NUH0047/SIN/08–B5G C T G G C A G G G C C C G G G T G A G C G C C NUH0086/SIN/08–B5G C T G G C A G G G C C C G G G T G A G C G C C NUH0083/SIN/08–B5G C T G G C A G G G C C C G G G T G A G C G C C NUH0085/SIN/08–B5G C T G G C A G G G C C C G G G T G A G C G C C NUH0043/SIN/08–B5G C T G G C A G G G C C C G G G T G A G C G C C NUH0037/SIN/08–B5G C T G G C A G G G C C C G G G T G A G C G C C NUH0012/SIN/08–B5G C T G G C A G G G C C C G G G T G A G C G C C NUH0075/SIN/08–C2G C C G G T A G A G C T C G G G T G A G G G C T NUH0013/SIN/08–C2G C C G G T A G A G C T C G G G T G A G G G C T HybridizaƟon probe G C T G G C A G G G C C T G G G T A A G T G C CFigure 2.Sequence alignment of 10outbreak EV71strains against the hybridization acceptor probe for real-time RT-PCR.The highlighted mismatches may explain the failure of detection of the EV71strains using this assay.The subgenogroups of the strains,and nucleotide positions at the 50(2496)and 30(2519)ends of the probe are shown.Y.Wu et al./International Journal of Infectious Diseases 14(2010)e1076–e1081e1078Figure 5highlights the differences,especially the non-polar to polar amino acid change within the SP55peptide.Therefore,the neutralizing antibodies of patients infected with previous EV71strains may not be able to recognize the 2008counterpart strains.In 2008,EV71epidemics were documented in many other countries in East Asia,but the behavior of the causative strains varied somewhat.For example,the predominant strain in the 2008China and Vietnam epidemics belonged to subgenogroup C4,16andNUH0037/SIN/08 NUH0012/SIN/08 NUH0083/SIN/08NUH0043/SIN/08NUH0085/SIN/08NUH0086/SIN/08NUH0049/SIN/08 NUH0047/SIN/08 2542-Yamagata-03 2716-Yamagata-03S110031-SAR-03 S19741-SAR-03 5536/SIN/00 S21082/SAR/004350/SIN/98MY16/1/SAR/970899-MAA-972604-AUS-74 2232-NY-777673-CT-87 7633-PA-872222-IA-88BrCr-CA/USA/701277S/VNM/05999T/VNM/051135T/VNM/05 SHZH98 H25-CHN-00 F2-CHN-00 2M/AUS/3/99 03750-MAA-97 NUH0075/SIN/08NUH0013/SIN/08KOR-EV71-01KOR-EV71-131M/AUS/12/00 2246-NY-87S11051/SAR/98 CVA16-G1099996699929999659989515446988556998984559999713562879876999099787199980.05B5B4B3B1B2C5C4C2C3C1AFigure 3.Dendrogram constructed based on the complete VP1gene sequences of 10outbreak EV71strains and selected known strains,elucidating B5and C2as the respectivemajor and minor EV71subgenogroups circulating during the 2008Singapore epidemic.The percentages of replicate trees in which the associated taxa are clustered together are shown next to the branches.Branch lengths are proportional to the number of nucleotide differences.Y.Wu et al./International Journal of Infectious Diseases 14(2010)e1076–e1081e1079exerted relatively high virulence culminating in numerous child fatalities.In contrast,the B5and C2strains of the 2008Singapore outbreak caused generally mild disease.VP1constitutes the major capsid protein and is considered to be an important factor that mediates viral pathogenesis.17To better understand differences in their VP1amino acid composition,the 10Singapore 2008strains were compared with two virulent strains from fatal cases,i.e.,Fuyang.Anhui.PRC/17.08/3from the 2008China outbreak and 5865/Sin/000009from the 2000Singapore outbreak.A disparity at position 22was noted,with R being replaced by Q or H in virulent strains,making it less basic.The Fuyang strain displayed an E to K substitution at position 98that could result in conformational change at the hydrophobic pocket of VP1.18Another mutation at amino acid 164of the virulent 2000Singapore strain was identified,but this was a conservative substitution.4.DiscussionHFMD has been a legally notifiable communicable disease in Singapore since October 1,2000,and the capture of epidemiologic data on HFMD has been effective since then.The 2008outbreak represents the largest HFMD epidemic in Singapore since the year 2000,with almost 30000cases,four patients with encephalitis,and one fatality.However,the actual number of HFMD cases may have been much higher given that most infections are asymptom-atic.There were two periods within which the number of infected cases increased significantly above the epidemic threshold,i.e.,a large peak from mid-March to the end of May (10927cases in weeks 12–22),and a smaller peak from early October to early December 2008(5391cases in weeks 42–49).However,in 2009,the number of HFMD cases did not exceed the epidemic threshold,and 17278cases were reported in 2009(without any encephalitis cases or deaths),compared with 29686for the whole of 2008(Figure 6).The specimens in our cohort were collected from patients with mild disease and without complications or sequelae,of whom only12%were hospitalized.Despite the relatively small sample size,this cohort is a subset representative of the pediatric HFMD cases in the community during the 2008Singapore outbreak,as the specimens were obtained during and after the peak of the epidemic curve based on distinct recruitment criteria.The most prevalent enterovirus infections associated with the 2008Singapore outbreak were attributed to CA6,followed by EV71and CA10.Both CA6and CA10are common etiologic agents of herpangina and have been prevalent in Japan since 2005,19,20and are less virulent but apparently more infectious than EV71.Consequently,the high transmissibility of HFMD during the 2008Singapore epidemic may be due to the dominance of CA6and CA10.In the autumn of 2008in Finland,CA6and CA10were also reported to be the predominant enteroviruses causing a large outbreak of HFMD with onychomad-esis.21,22The similar clinical presentations of HFMD and herpangina make it difficult for physicians to accurately diagnose the two disease entities.Most patients infected by non-EV71and non-CA16enteroviruses presented fewer than five papules,whereas the significant majority of EV71and CA16patients exhibited more than five papules.From a general clinical perspective,it is suspected that patients with fewer than five papules may actually be suffering from herpangina rather than HFMD.Hence,we postulate that this outbreak comprised a mixture of HFMD as well as herpangina cases.Moreover,the percentage of patients who had fever was significantly different between patients with EV71and those with non-EV71infections.Besides CA6and CA10,another major contributor was EV71,which accounted for more than one third of enterovirus-positive cases.VP1-based phylogenetic analysis revealed two EV71subgenogroups,namely B5and C2.There have been two major lineages (B and C)circulating during HFMD outbreaks in Southeast Asia since 1997,with five subgenogroups under genogroup B.B1and B2were identified throughout the world during the 1970s andEV71B5strainCollecƟon date (2008)719373427439604644658756820832866NUH0049/SIN/087Apr T C A T T C A T T C A A NUH0047/SIN/087Apr T C A T T C A T T C A A NUH0086/SIN/086May T C A T T C A T T C A A NUH0083/SIN/0815May C T G T C T A T C C A A NUH0085/SIN/083Jun T T G C T C A C C C A A NUH0043/SIN/0812Jun T T G C T C G T C C A A NUH0037/SIN/0814Aug C T G T C T A T C T G G NUH0012/SIN/0815AugT T G T C T A T C C A AFigure 4.Alignment of VP1nucleotides of eight EV71strains belonging to subgenogroup B5according to the time of specimen receipt.The disparities at 12different positions highlight the evolution of the VP1regions of B5strains during the course of the large epidemic.For example,five disparities at positions 644,658,820,832,and 866emerged in strains that were obtained later in the mon disparities at nucleotides 19,373,and 756were also identified only in strains detected later.SP12(34–48)VSSHRLDTG E VPALQ C 2008strains VSSHRLDTG (K/E)VPALQ A SP55(163–177)P E SRESLAWQTATNP C 2008strains P D SRESLAWQTATNP S SP70(208–222)YPTFGEHKQEKDLEY C 2008strainsYPTFGEHKQEKDLEY GFigure 5.Amino acid sequence variations within the VP1neutralizing antibody epitopes SP12,SP55,and SP70of the 2008outbreak EV71strains.Figure 6.Weekly incidence of hand,foot and mouth disease in Singapore during 2008and 2009.The dotted line depicts the epidemic threshold of 665cases (.sg/mohcorp/statisticsweeklybulletins.aspx ).Y.Wu et al./International Journal of Infectious Diseases 14(2010)e1076–e1081e10801980s.B3and B4later emerged as the dominant subgenogroups in Australia,Malaysia,and Singapore.23–25Subgenogroup B5wasfirst identified in Japan in2003,and replaced the previous dominant strain within a short period of time.26In Singapore,the transition of the predominant EV71subgenogroup from B4in200027to B5in 2008correlated with the reduced fatality rate from 7/4000to 1/ 30000.Replacement of subgenogroups was also witnessed in Taiwan and Sarawak in2008.28Subgenogroup C2was initially identified in Taiwan,being responsible for the largest ever HFMD outbreak in1998.In the2008Singapore outbreak,C2subge-nogroup accounted for only one-fifth of EV71-positive samples, relegating it to a minor role.Nevertheless,this reiterates that multiple genetic lineages of EV71circulate endemically in the Singapore population all year round.The complete nucleotide sequence analyses of whole EV71 genomes facilitate the characterization of circulating strains at the genetic level and of their predicted viral proteins.Interactions between viruses and their hosts play critical roles in virus evolution of structural and non-structural genes.Quite a number of VP1mutations were observed in the10studied EV71strains in comparison with known sequenced strains.This reflects the relatively high mutation rate of EV71,which helps it to escape human immune surveillance.The considerable mutation rate of the EV71capsid protein may also have implications on future vaccine development.29Furthermore,disparities were also dis-covered in other non-structural and non-coding regions such as the 3D polymerase and50UTR(data not shown).Finally,a safe and effective vaccine against EV71is certainly warranted in view of its potential neurovirulence and its role in HFMD epidemics of recurring frequency with resultant fatalities in Asia,as well as other parts of the world.15Conflict of interestWe declare that we have no conflict of interest with respect to this study.Financial supportThis study was supported by a grant from the Academic Research Fund,Ministry of Education,Singapore(R178-000-146-112).AcknowledgementsWe thank the patients and their parents who participated in this study,our clinical colleagues at the National University Hospital,Singapore for obtaining specimens,and u for technical assistance.References1.McMinn PC.An overview of the evolution of enterovirus71and its clinical andpublic health significance.FEMS Microbiol Rev2002;26:91–107.2.Pe´rez-Ve´lez CM,Anderson MS,Robinson CC,McFarland EJ,Nix WA,PallanschMA,et al.Outbreak of neurologic enterovirus type71disease:a diagnostic challenge.Clin Infect Dis2007;45:950–7.3.Qiu J.Enterovirus71infection:a new threat to global public health?LancetNeurol2008;7:868–9.4.Singh S,Chow VT,Phoon MC,Chan KP,Poh CL.Direct detection of enterovirus71(EV71)in clinical specimens from a hand,foot,and mouth disease outbreak in Singapore by reverse transcription-PCR with universal enterovirus and EV71-specific primers.J Clin Microbiol2002;40:2823–7.5.Leong PW,Liew K,Lim W,Chow VT.Differential display RT-PCR analysis ofenterovirus-71-infected rhabdomyosarcoma cells reveals mRNA expression responses of multiple human genes with known and novel functions.Virology 2002;295:147–59.6.Leong WF,Chow VT.Transcriptomic and proteomic analyses of rhabdomyosar-coma cells reveal differential cellular gene expression in response to enterovi-rus71infection.Cell Microbiol2006;8:565–80.7.Tan EL,Chow VT,Kumarasinghe G,Lin RT,Mackay IM,Sloots TP,et al.Specificdetection of enterovirus71directly from clinical specimens using real-time RT-PCR hybridization probe assay.Mol Cell Probes2006;20:135–40.8.Rotbart HA.Enzymatic RNA amplification of the enteroviruses.J Clin Microbiol1990;28:438–42.9.Zoll GJ,Melchers WJ,Kopecka H,Jambroes G,van der Poel HJ,Galama JM.General primer-mediated polymerase chain reaction for detection of entero-viruses:application for diagnostic routine and persistent infections.J Clin Microbiol1992;30:160–5.10.Oberste MS,Maher K,Kilpatrick DR,Flemister MR,Brown BA,Pallansch MA.Typing of human enteroviruses by partial sequencing of VP1.J Clin Microbiol 1999;37:1288–93.11.Singh S,Chow VT,Chan KP,Ling AE,Poh CL.RT-PCR,nucleotide,amino acid andphylogenetic analyses of enterovirus type71strains from Asia.J Virol Methods 2000;88:193–204.12.Brown BA,Oberste MS,Alexander JP,Kennett ML,Pallansch MA.Molecularepidemiology and evolution of enterovirus71strains isolated from1970to 1998.J Virol1999;73:9969–75.13.Foo DG,Alonso S,Phoon MC,Ramachandran NP,Chow VT,Poh CL.Identificationof neutralizing linear epitopes from the VP1capsid protein of enterovirus71 using synthetic peptides.Virus Res2007;125:61–8.14.Foo DG,Alonso S,Chow VT,Poh CL.Passive protection against lethal enterovirus71infection in newborn mice by neutralizing antibodies elicited by a synthetic peptide.Microbes Infect2007;9:1299–306.15.Ho SY,Chua SQ,Foo DG,Locht C,Chow VT,Poh CL,et al.Highly attenuatedBordetella pertussis strain BPZE1as a potential live vehicle for delivery of heterologous vaccine candidates.Infect Immun2008;76:111–9.16.Huemer HP,Ortner B,Huang CW,Schmid D,Mutz I,Wewalka G,et al.IsolatingAsian enterovirus71subgenogroup C4in two Austrian clinical samples from 2004.Euro Surveill2008;13.pii:18922.l SK,Kumar P,Yeo WM,Kar-Roy A,Chow VT.The VP1protein of humanenterovirus71self-associates via an interaction domain spanning amino acids 66–297.J Med Virol2006;78:582–90.18.Chen TC,Liu SC,Huang PN,Chang HY,Chern JH,Shih SR.Antiviral activity ofpyridyl imidazolidinones against enterovirus71variants.J Biomed Sci 2008;15:291–300.19.Yamashita T,Ito M,Taniguchi A,Sakae K.Prevalence of coxsackievirus A5,A6,and A10in patients with herpangina in Aichi Prefecture,2005.Jpn J Infect Dis 2005;58:390–1.20.Sano T,Saito T,Kondo M,Watanabe S,Onoue Y,Konnai M,et al.Enterovirusdetection status of patients with herpangina and hand,foot and mouth disease in epidemic season2007,Kanagawa Prefecture,Japan.Jpn J Infect Dis 2008;61:162–3.21.Osterback R,Vuorinen T,Linna M,Susi P,Hyypia¨T,Waris M.Coxsackievirus A6and hand,foot,and mouth disease,Finland.Emerg Infect Dis2009;15:1485–8.22.Blomqvist S,Klemola P,Kaijalainen S,Paananen A,Simonen ML,Vuorinen T,et al.Co-circulation of coxsackieviruses A6and A10in hand,foot and mouth disease outbreak in Finland.J Clin Virol2010;48:49–54.23.McMinn P,Lindsay K,Perera D,Chan HM,Chan KP,Cardosa MJ.Phylogeneticanalysis of enterovirus71strains isolated during linked epidemics in Malaysia, Singapore,and Western Australia.J Virol2001;75:7732–8.24.Cardosa MJ,Perera D,Brown BA,Cheon D,Chan HM,Chan KP,et al.Molecularepidemiology of human enterovirus71strains and recent outbreaks in the Asia-Pacific region:comparative analysis of the VP1and VP4genes.Emerg Infect Dis 2003;9:461–8.25.Shimizu H,Utama A,Yoshii K,Yoshida H,Yoneyama T,Sinniah M,et al.Enterovirus71from fatal and nonfatal cases of hand,foot and mouth disease epidemics in Malaysia,Japan and Taiwan in1997–1998.Jpn J Infect Dis 1999;52:12–5.26.Mizuta K,Abiko C,Murata T,Matsuzaki Y,Itagaki T,Sanjoh K,et al.Frequentimportation of enterovirus71from surrounding countries into the local com-munity of Yamagata,Japan,between1998and2003.J Clin Microbiol 2005;43:6171–5.27.Singh S,Poh CL,Chow plete sequence analyses of enterovirus71strainsfrom fatal and non-fatal cases of the hand,foot and mouth disease outbreak in Singapore(2000).Microbiol Immunol2002;46:801–8.28.Huang SW,Hsu YW,Smith DJ,Kiang D,Tsai HP,Lin KH,et al.Reemergence ofenterovirus71in2008in Taiwan:dynamics of genetic and antigenic evolution from1998to2008.J Clin Microbiol2009;47:3653–62.29.Bible JM,Pantelidis P,Chan PK,Tong CY.Genetic evolution of enterovirus71:epidemiological and pathological implications.Rev Med Virol2007;17:371–9.Y.Wu et al./International Journal of Infectious Diseases14(2010)e1076–e1081e1081。
Self-Templated Synthesis of Nanoporous CdS Nanostructures
Self-Templated Synthesis of Nanoporous CdS Nanostructures for Highly Efficient Photocatalytic Hydrogen Production underVisible LightNingzhong Bao,*,†Liming Shen,†Tsuyoshi Takata,and Kazunari Domen Department of Chemical System Engineering,School of Engineering,The Uni V ersity of Tokyo,7-3-1Hongo,Bunkyo-Ku,Tokyo113-8656,JapanRecei V ed October11,2007.Re V ised Manuscript Recei V ed No V ember5,2007Nanoporous CdS nanostructures,including nanosheets and hollow nanorods,have been prepared by a two-step aqueous route,which consists of afirst precipitation of nanoporous Cd(OH)2intermediates and a subsequent S2-/OH-ion-exchange conversion of the obtained Cd(OH)2used as template either to nanoporous CdS nanosheets with sizes up to60nm and an average thickness of about9nm or to CdS hollow nanorods with lengths up to30nm and outer diameters in the range7–14nm.The obtained CdS nanostructures containing nanopores with diameters of∼3nm exhibit a very large BET surface area of about112.8m2g-1.A very high hydrogen yield of about4.1mmol h-1under visible light irradiation (λg420nm),corresponding to the highest apparent quantum yield of about60.34%measured at420 nm so far reported,has been attained over the obtained nanoporous CdS nanostructures loaded with monodisperse3–5nm Pt nanocrystals,which is due to an efficient charge separation,a fast transport of the photogenerated carriers,and a fast photochemical reaction at the CdS/electrolyte interface.The photocatalytic reaction conditions,such as the Pt-loading content,the amount of catalyst,and the concentration of sacrificial regents,have been optimized.1.IntroductionHydrogen,an attractive sustainable clean energy source, is currently obtained from nonrenewable natural gas,petro-leum,and coal but could in principle be generated from the renewable resource of water.1–7The photosplitting of water is an attractive environmental-friendly method,which offers a way of capturing available solar energy and converting it into valuable hydrogen.8–11However,most of developed photocatalysts capable of splitting water are mixed transition metal oxides with wide band gaps,which can only take advantage of ultraviolet irradiation constituting only4%of the incoming solar energy.Considerable efforts have been focused on developing visible-light-driven photocatalysts,such as limited mixed oxides,12–14(oxy)nitrides,15–21and (oxy)sulfdes,22,23capable of using the less energetic but more abundant visible light(λg420nm)accounting for about 43%of the solar spectrum.Sulfides,such as CdS,ZnS-CuInS2solid solution,etc., have narrow band gaps and valence bands at relatively negative potentials,which offer them good visible-light-driven photocatalytic activities in the presence of sacrificial reagents.24–32The usage of sacrificial reagents,such as S2-,*Corresponding author:Tel205-348-5041;fax205-348-9104;e-mail nzhbao@.†Present address:Center for Materials for Information Technology(MINT), The University of Alabama,Tuscaloosa,AL35487.(1)Maeda,K.;Teramura,K.;Lu,D.;Saito,N.;Inoue,Y.;Domen,K.Angew.Chem.,Int.Ed.2006,45,7806.(2)Maeda,K.;Teramura,K.;Lu,D.;Takata,T.;Saito,N.;Inoue,Y.;Domen,K.Nature(London)2006,440,295.(3)Ishikawa,A.;Takata,T.;Kondo,J.N.;Hara,M.;Kobayashi,H.;Domen,K.J.Am.Chem.Soc.2002,124,13547.(4)Kim,H.G.;Borse,P.H.;Choi,W.;Lee,J.S.Angew.Chem.,Int.Ed.2005,44,4585.(5)Zou,Z.;Ye,J.;Sayama,K.;Arkawa,H.Nature(London)2001,414,625.(6)Gratzel,M.Nature(London)2001,414,338.(7)Khan,S.U.M.;Al-shahry,M.;Ingler,W.B.,Jr Science2002,279,2243.(8)Bard,A.J.;Fox,M.A.Acc.Chem.Res.1995,28,141.(9)Khaselev,O.;Turner,J.A.Science1998,280,425.(10)Fujishima,A.;Honda,K.Nature(London)1972,238,37.(11)Kato,H.;Asakura,K.;Kudo,A.J.Am.Chem.Soc.2003,125,3082.(12)Kudo,A.;Omori,K.;Kato,H.J.Am.Chem.Soc.1999,121,11459.(13)Kato,H.;Kudo,A.J.Phys.Chem.B2001,105,4285.(14)Kato,H.;Kudo,A.J.Phys.Chem.B2002,106,5029.(15)Chun,W.;Ishikawa,A.;Fujisawa,H.;Takata,T.;Kondo,J.N.;Hara,M.;Kawai,M.;Matsumoto,Y.;Domen,K.J.Phys.Chem.B2003, 107,1798.(16)Kasahara,A.;Nukumizu,K.;Takata,T.;Kondo,J.N.;Hara,M.;Kobayashi,H.;Domen,K.J.Phys.Chem.B2003,107,791. (17)Hara,M.;Chiba,E.;Ishikawa,A.;Takata,T.;Kondo,J.N.;Domen,K.J.Phys.Chem.B2003,107,13441.(18)Maeda,K.;Takata,T.;Hara,M.;Saito,N.;Inoue,Y.;Kobayashi,H.;Domen,K.J.Am.Chem.Soc.2005,127,8286.(19)Maeda,K.;Teramura,K.;Lu,D.;Saito,N.;Inoue,Y.;Domen,K.Angew.Chem.,Int.Ed.2006,45,7806.(20)Lee,Y.;Terashima,H.;Shimodaira,Y.;Teramura,K.;Hara,M.;Kobayashi,H.;Domen,K.;Yashima,M.J.Phys.Chem.C2007,111, 1042.(21)Ito,S.;Thampi,K.R.;Comte,P.;Liska,P.;Gratzel,M.Chem.Commun.2005,268.(22)Ishikawa,A.;Yamada,Y.;Takata,T.;Kondo,J.N.;Hara,M.;Kobayashi,H.;Domen,K.Chem.Mater.2003,15,4442.(23)Ishikawa,A.;Takata,T.;Matsumura,T.;Kondo,J.N.;Hara,M.;Kobayashi,H.;Domen,K.J.Phys.Chem.B2004,108,2637. (24)Tsuji,I.;Kato,H.;Kobayashi,H.;Kudo,A.J.Am.Chem.Soc.2004,126,13406.(25)Tsuji,I.;Kato,H.;Kudo,A.Angew.Chem.,Int.Ed.2005,44,3565.(26)Tsuji,I.;Kato,H.;Kudo,A.Chem.Mater.2006,18,1969.(27)Tsuji,I.;Kato,H.;Kobayashi,H.;Kudo,A.J.Phys.Chem.B2005,109,7323.110Chem.Mater.2008,20,110–11710.1021/cm7029344CCC:$40.75 2008American Chemical SocietyPublished on Web12/06/2007SO32-,etc.,prevents sulfide photocatalysts from the anodic photocorrosion by providing sacrificial electron donors to consume the photogenerated holes.At the same time,it promotes hydrogen evolution through making up half of the water-splitting reaction.33–35Because the extraction products, such as sulfides,sulfites,etc.,of fossil energy resources now being produced in large quantities are undesirable and polluting byproduct in hydrogenation andflue-gas desulfu-rization processes at chemical plants,the photocatalytic reaction yielding valuable hydrogen energy over sulfides from aqueous solution containing S2-and SO32-is very attractive both for achieving practical photocatalytic hydrogen production using sunlight and in solving the environmental problem caused by the petrochemical industries.The CdS photocatalysts with good photocatalyic activity process the combination of materials-related characteristics in particularly requiring hexagonal crystal structure,good crystallinity,large surface area,good dispersity,short bulk-to-surface diffusion distance for e-and h+,and monodisperse cocatalyst-sulfide surface nanostructures,which contributes to an efficient charge separation,a fast transport of the photogenerated carriers,and a fast photochemical reaction at the photocatalyst/electrolyte interface.36–38The Ag2S-activated ZnS-doped colloidal CdS nanocrystals are the most active CdS ever reported,exhibiting a very high quantum yield of37%at450nm,which is due to a very large surface area of>100m2g-1.35However,it suffers from disadvan-tages of serious particle agglomeration and unstability. Compared with the above Ag2S-activated ZnS-doped col-loidal CdS nanocrystals,a relatively higher quantum yield of25%at450nm has been achieved over platinized pure CdS powder which is stable but has a low specific surface area of<6.7m2g-1.Therefore,there is large room for a further improvement of the photocatalytic activity of pure CdS by increasing the surface area.39Nanoporous sheetlike/hollow CdS nanostructures can be prepared in relatively large sizes up to dozens of nanometers, which decreases the particle agglomeration occurring in many colloidal nanocrystals.Meanwhile,at least the thickness of the sheetlike nanostructures or the wall of hollow nanostruc-tures has the size down to several nanometers.Electrons and holes generated inside the CdS are able to quickly migrate to the surface of CdS,passing a shorter distance of either half of the thickness or near to the nanopore wall,which greatly decreases the bulk e-/h+recombination.At the same time,a quick surface e-/h+separation,a fast interfacial charge carrier transfer,and an easy charge carrier trapping are achieved,which is due to the factor that the large surface area and the nanoporous structure ensure the strong abilities to interact with ions,atoms,and molecules not only on the outer surface but also throughout the whole nanostructures.40–44 As a result,the nanoporous sheetlike/hollow CdS nanostruc-tures are capable of exhibiting highly visible-light-driven photocatalytic activity for hydrogen production.45–47On the other hand,there still exist several disadvantages for the nanoporous/hollow nanostructures.First,the synthesis of materials containing nanopores and hollow structures often requires synthetic templates,and thus a simple,direct,and low-cost synthesis approach needs to be developed.40Second, microporous solids,such as zeolites,etc.,consist of three-dimensional crystalline frameworks and interpenetrating small micropore channels(<1nm in diameter),which cannot allow thefixation of large molecules,the reduction of the diffusion restriction of reactants,reactions involving bulky molecules,and in particular for the aqueous heterogeneous photocatalytic reaction involving sacrificial reagents.48–50Third, the majority of mesoporous(2–50nm diameter)materials, in particular for nonoxides,cannot be synthesized in crystal-line form,and only several limited semicrystalline mesopo-rous materials have been reported.51–53Therefore,the de-velopment of a simple,mild,and effective synthetic method to directly prepare crystalline nanoporous CdS nanostructures is a key step for achieving highly efficient photocatalytic hydrogen production under visible light.In this study,we report a simple,aqueous solution route for the large-scale preparation of crystalline nanoporous CdS nanostructures at room temperature under air condition using air-insensitive inorganic reactants of CdCl2·2.5H2O and Na2S·9H2O.The synthesis procedure involves an initial precipitation of nanoporous Cd(OH)2nanosheets and nano-rods in aqueous solution and a subsequent S2-/OH-ion-exchange conversion of the obtained Cd(OH)2intermediates to CdS.The obtained CdS nanostructures loaded with monodisperse uniform Pt nanocrystals exhibit the highest apparent quantum yield of about60.34%at420nm for photocatalytic hydrogen production in the presence of sacrificial reagents of Na2SO3and Na2S,showing the potential for practical photocatalytic hydrogen production(28)Shangguan,W.;Yoshida,A.J.Phys.Chem.B2002,106,12227.(29)Fujishiro,Y.;Uchida,S.;Sato,T.Int.J.Inorg.Mater.1999,1,67.(30)Lei,Z.;Ma,G.;Liu,M.;You,W.;Yan,H.;Wu,G.;Takata,T.;Hara,M.;Domen,K.;Li,C.J.Catal.2006,237,322.(31)Tricot,Y.M.;Fendler,J.H.J.Am.Chem.Soc.1984,106,2475.(32)Uchihara,T.;Matsumura,M.;Yamamoto,A.;Tsubomura,H.J.Phys.Chem.1989,93,5870.(33)Bulher,N.;Meier,K.;Reber,J.F.J.Phys.Chem.1984,88,3261.(34)Reber,J.F.;Meier,K.J.Phys.Chem.1984,88,5903.(35)Reber,J.F.;Rusek,M.J.Phys.Chem.1986,90,824.(36)Kudo,A.;Kato,H.;Tsuji,I.Chem.Lett.2004,33,1534.(37)Fox,M.A.;Dulay,M.T.Chem.Re V.1995,83,341.(38)Bao,N.;Shen,L.;Takata,T.;Domen,K.;Gupta,A.;Yanagisawa,K.;Grimes,A.C.J.Phys.Chem.C,2007,111,17527.(39)Matsumura,M.;Furukawa,S.;Saho,Y.;Tsubomura,H.J.Phys.Chem.1985,89,1329.(40)Davis,M.E.Nature(London)2002,417,813.(41)Bao,N.;Shen,L.;Yanagisawa,K.J.Phys.Chem.B2004,108,16739.(42)Bao,N.;Feng,X.;Yang,Z.;Shen,L.;Lu,X.En V iron.Sci.Technol.2004,38,2729.(43)Lu,D.;Hitoki,G.;Katou,E.;Kondo,J.N.;Hara,M.;Domen,K.Chem.Mater.2004,16,1603.(44)Ito,S.;Thampi,K.R.;Comte,P.;Liska,P.;Gratzel,M.Chem.Commun.2005,268.(45)Bao,N.;Shen,L.;Takata,T.;Lu,D.;Domen,K.Chem.Lett.2006,35,318.(46)Zheng,N.;Bu,X.;Vu,H.;Feng,P.Angew.Chem.,Int.Ed.2005,44,5299.(47)Hu,J.;Ren,L.;Guo,Y.;Liang,H.;Cao,A.;Wan,L.;Bai,C.Angew.Chem.,Int.Ed.2005,44,1269.(48)Corma,A.Chem.Re V.1997,97,2373.(49)Soler-Illia,G.J.;de,A.A.;Sanchez,C.;Lebeau,B.;Patarin,J.Chem.Re V.2002,102,4093.(50)Ying,J.Y.;Mehnert,C.P.;Wong,M.S.Angew.Chem.,Int.Ed.1999,38,56.(51)Katou,T.;Lee,B.;Lu,D.;Kondo,J.N.;Hara,M.;Domen,K.Angew.Chem.,Int.Ed.2003,42,2382.(52)Yang,P.;Zhao,D.;Margolese,D.I.;Chmelka,B.F.;Stucky,G.D.Nature(London)1998,396,152.(53)Crepaldi,E.L.;Soler-Illia,G.J.;de,A.A.;Grosso,D.;Cagnol,F.;Ribot,F.;Sanchez,C.J.Am.Chem.Soc.2003,125,9970.111Chem.Mater.,Vol.20,No.1,2008Self-Templated Synthesis of Nanoporous CdSusing sunlight.The present synthesis strategy could be a general method for other binary sulfides.2.Experimental Section2.1.Preparation and Characterization of Photocatalysts.Ina typical synthesis,the white Cd(OH)2intermediate was precipitated by adding10mL of0.1M CdCl2·2.5H2O aqueous solution to100 mL of0.1M NaOH aqueous solution under stirring for2min at room temperature.The obtained white Cd(OH)2intermediate was then converted to yellow CdS by adding10mL of0.1M Na2S·9H2O to the above solution under continuous stirring for5 min.The obtained CdS precipitate was collected,washed with distilled water for several times,andfinally dried overnight in an oven at70°C.The obtained CdS were investigated using a combination of characterization technologies including N2Sorption (BEL,BELsorp-mini),transmission electron microscopy(TEM, coupled with high-resolution(HR)and energy dispersive spectros-copy(EDS),Tecnai F-20),scanning electron microscopy(Hitachi S-4700),X-ray diffraction(XRD,Rigaku RINT-UltimaIII;Cu K R), and ultraviolet–visible diffuse reflectance spectra(UV–vis DRS, JASCO V-560).2.2.Photocatalytic Reaction.Photocatalytic reactions were carried out in a closed gas circulation and evacuation systemfitted with a top window Pyrex cell.Prior to the photocatalytic reaction, the CdS powders were uniformly platinized by a0.5h photore-duction of1wt%H2PtCl6with UV light irradiated using a450W high-pressure Hg lamp.Approximately0.15g of Pt-loaded CdS photocatalyst was dispersed by a magnetic stirrer in200mL of aqueous solution containing0.35M Na2SO3and0.25M Na2S as sacrificial reagents.The photocatalysts were irradiated with visible light(λg420nm)using a cutofffilter from a300W Xe lamp. The temperature of the reactant solution was maintained at room temperature by providing aflow of cooling water during the photocatalytic reaction.The amount of hydrogen evolved was determined with online gas chromatography.The apparent quantum yield was measured under the same photocatalytic reaction except for the wavelength of irradiation light. The hydrogen yields of10h photocatalytic reaction in one continuous reaction under visible light with different wavelengths of420,470,500,520,560,and600nm were measured.Apparent quantum yields at different wavelengths were calculated by the following function.The band-pass and cutofffilters and a photo-diode were used in measurement.apparent)number of photons used to generate H2 total number of photons underirradiation at fixed wavelengthPhotocatalytic hydrogen production by irradiating suspensions containing platinized CdS and the electrolytes of S2-and SO32–has shown very high efficiency.The addition of SO32-can efficiently suppress the formation of disulfide(S22-)ions,allowing hydrogen to evolve at a high rate.Chemical analysis has been used to confirm the photodegradation products of sacrificial regents of S2-and SO32-.SO42-ions in the presence of SO32-and S2O32-ions were analyzed by precipitation with BaCl2solution.The excess of BaCl2was then titrated with EDTA solution.After SO32-ions were separated byflushing N2through the acidified solution,the concentration of S22-ions was determined by oxidation with a hot solution of K2Cr2O7.Analysis of S2O32-ions in the presence of S2-and SO32-ions was carried out iodometrically after separating S2-ions by precipitation with zinc acetate in an alkane medium and masking SO32-with formaldehyde.3.Results and Discussion3.1.Characterizations and Properties of Photocatalysts. XRD patterns of the Cd(OH)2intermediates and the CdS products,together with the standard diffraction patterns of Cd(OH)2and hexagonal CdS,are depicted in Figure1.All the different peaks of products are well indexed either as the Cd(OH)2(JCPDS card No.31-0228)in Figure1a or as the hexagonal CdS(JCPDS card No.41-1049)in Figure1b. Figure1b shows three very broadening diffraction peaks attributed to the(002),(110),and(112)planes of a hexagonal phase of CdS.The average grain sizes,calculated from the full width at half-maximum(fwhm,in radians),of the(101) line for the Cd(OH)2intermediates and the(101)line for the CdS products using the Scherrer formula are12.4and 3.5nm,respectively.The morphology,structure,and composition of the ob-tained products have been investigated by TEM,HRTEM, SEM,and EDS.The Cd(OH)2intermediates(Figure1)are composed of nanosheets of dozens of nanometers in size and nanorods of9–14nm in diameter and40–60nm in length. The HRTEM image(Figure2a2)of a typical individual Cd(OH)2nanosheet shows clear crystal lattices with short-range continuous size up to5nm,indicating the polycrys-talline nature of the nanosheets.HRTEM image(Figure2a3) of a typical individual nanorod clearly shows large-scale continuous crystal lattices with an interplanar distance of about0.3nm corresponding to the(100)plane,confirming the single crystalline nature of the nanorods.Nanopores with diameter of∼3nm are observed within both nanosheets and nanorods.The Cd(OH)2intermediates have been converted to CdS via a S2-ion-exchange reaction.As shown in Figure2b1, the shape and size of the obtained CdS nanostructures are very close to those of the Cd(OH)2intermediates.AHRTEM Figure1.XRD patterns of(a)Cd(OH)2intermediates and(b)as-made hexagonal CdS and standard XRD patterns of Cd(OH)2(JCPDS No.31-0228)and hexagonal CdS(JCPDS No.41-1049).112Chem.Mater.,Vol.20,No.1,2008Bao et al.image (Figure 2b2)of an individual CdS nanosheet shows many random arranged crystal lattices with short-range continuous sizes,which indicates a polycrystalline nature of the CdS nanosheets.Figure 2b3shows HRTEM image of two CdS hollow nanorods with inner diameters of 2.8and 6.2nm.Both of the CdS hollow nanorods have clear,continuous crystal lattices,indicating the single crystalline nature of the obtained hollow CdS nanorods.A penetrable nanopore,marked with a red arrow in Figure 2b3,is observed at one end of a hollow nanorod.The obtained CdS nano-structures with special nanopores and hollow structures have never been reported.Pt nanocrystals have been uniformly photodeposited on the obtained CdS nanostructures.No morphologic and structural differences have been observed between the as-made CdS nanostructures (Figure 2b1)and the Pt-loaded CdS nanostructures (Figure 2c1).The EDS spectra (not shown)of the Pt-loaded nanoporous CdS nanostructures show strong Cd and S signals with Cd:S ratios of 1:1.03,which indicates the Cd(OH)2intermediates have been totally converted to CdS.Furthermore,Pt signal with 8%content (the atom content to Cd)was also detected.As shown in Figure 2c2,c3,monodisperse Pt nanocrystals with uniform size of about 3nm have been deposited on the surface of CdS nanostruc-tures.HRTEM image (Figure 2c4)of a CdS nanosheet shows two different clear crystal lattices with interplanar distances of 0.34nm of the CdS(002)plane and 0.23nm of the Pt(111)plane.More stereomicrographs of the Pt-loaded CdS nano-structures were obtained using SEM.As shown in Figure 2c5,either nanosheets with size up to 50nm and thickness of around 9nm or the hollow nanorods with diameter of around 10nm and length up to 40nm were observed in the CdS products covering with monodisperse Pt nanocrystals (see Figure 2c6),in good agreement with the above TEM results.Textural property of the obtained nanoporous CdS nano-structures has been determined by N 2adsorption/desorption isotherms and corresponding pore size distribution.AsshownFigure 2.TEM,HRTEM,and SEM images of the products formed at various synthestic steps.(a1-a3)TEM images of Cd(OH)2intermediates:(a1)TEM image of typical Cd(OH)2intermediates;(a2)and (a3)respectively are TEM images of an individual nanosheet and nanorod both containing nanopores marked with red arrows.(b1-b3)TEM and HRTEM images of nanoporous nanosheets and hollow nanorods of polycrystalline CdS:(b1)TEM image of typical CdS nanostructures;(b2)and (b3)respectively are HRTEM images of individual nanosheet and hollow nanorod of polycrystalline CdS.(c1-c6)TEM,HRTEM,and SEM images of Pt-loaded CdS nanosheets and hollow nanorods:(c1)TEM image of typical Pt-loaded CdS nanostructures;(c2,c3)magnified TEM images of CdS nanosheets showing monodisperse Pt nanocrystals on the surface;(c4)HRTEM image of an individual Pt-loaded CdS nanosheet,showing crystal lattices of both CdS and Pt;(c5,c6)SEM images of Pt-loaded CdS nanostructures.Some Pt nanocrystals have been marked with blue arrows in parts c1,c2,and c6.113Chem.Mater.,Vol.20,No.1,2008Self-Templated Synthesis of Nanoporous CdSin Figure 3,two well-defined steps at P /P 0of 0.5–0.9and 0.9–1were observed on the N 2adsorption/desorption iso-therms.The former gives a typical type IV isotherm with a clear H1-type hysteresis loop which is characteristic of mesoporous materials.The latter of the uptake at high pressure is associated with the empty spaces between nanoparticles.A relatively narrow pore size distribution with maximum at around 3.3nm is also shown in the inset of Figure 3,in agreement with the TEM results.Other typical textural parameters of the as-made CdS nanostructures are BET surface area of 112.8m 2g -1and pore volume of 0.38cm 3g -1,which are very high for crystalline semiconductor sulfides.Figure 4show the UV–vis diffuse reflectance spectra of the as-made CdS and the Pt-loaded CdS nanostructures,corresponding to a band gap of about 2.25eV,which are comparable to the measured value for the standard bulk CdS sample (about 2.4eV).Generally,there exist critical sizes on nanoscale for semiconductors exhibiting a blue shift in the absorption spectrum.As for the CdS,a blue shift can be observed in the absorption spectrum when the size of CdS is smaller than 5nm.54,55Although the structural units of the present CdS nanostructures have a relatively fine size of about 5nm,the macroscopic size of each individualnanostructure is larger than decades of nanometers,which could be the reason why the absorption spectrum does not exhibit a blue shift.3.2.Photocatalytic Activity of Nanoporous CdS Nano-structures.Figure 5a shows the photocatalytic activity of the Pt-loaded CdS nanostructures decreased slightly after four run reactions in one continuous reaction.The initially photocatalytic hydrogen yield attained4.1mmol h -1.After the fourth reaction runs,the photocatalyst retained 80%of the initial photocatalytic activity under visible light irradiation and reached a stable hydrogen yield of 3.1mmol h -1.Figure 5b shows an action spectrum for hydrogen evolution from an aqueous solution containing both 0.35M Na 2SO 3and 0.25M Na 2S over the Pt(10wt %)-loaded photocatalysts of CdS nanostructures.The onset of the action spectrum agrees well with that of the diffuse reflection spectrum.Hydrogen evolution stopped by absorption in the tail region (>600nm),revealing that the visible-light response of the photocatalyst is due to the band-gap transition between the valence and conduction bands.The quantum yield of the Pt-loaded CdS nanostructures is 60.34%at 420nm,which is the highest apparent quantum yield so far reported.Previous results reported active Ag 2S activated CdS doping with ZnS showing a high quantum yield of 37%at 450nm,but the materials are less stable than platinized pure CdS powders,giving a quantum yield of 25%at 450nm at 60°C.32,33The apparent quantum yield decreased with increasing the active wave-length until the lowest value of about 0.2%at 600nm.Hydrogen evolution did not appear by absorption in the region of >600nm because only photons of a wavelength shorter than 600nm can induce photochemical reactions.The change tredency of the quantum yields at different waver-lengths should agree with that of the UV–vis DSR.24,33The quantum yield at 450nm should be close to that obtained at 420nm because of the agreeable absorbance at both 420and 450nm.The present results show that the quantum yield at 450nm is about 20%lower than 60.34%at 420nm,which is due to the fact that,in the present study,we measured the quantum yields at 420,470,500,520,560,and 600nm in one continuous reaction,and the reaction at each wavelength continued for 10h,which caused the decreased hydrogen evolution rate because of both the contamination of the platinum oxidized at the CdS surface by sulfurous products to form platinum sulfide and an increasing concentration of(54)Nakanishi,T.;Ohtani,B.;Uosaki,K.J.Phys.Chem.B 1998,102,1571.(55)Wang,Y.;Suna,A.;Mahler,W.;Kosowski,R.J.Chem.Phys.1987,87,7315.Figure 3.Nitrogen adsorption/desorption isotherms and pore diameter distribution (the inset,calculated from the adsorption branch using the BJH method)of as-made CdSnanostructures.Figure 4.UV–vis diffuse reflectance spectrum of (a)as-made hexagonal CdS and (b)10wt %Pt-loaded CdSnanostructures.Figure 5.(a)Time course of hydrogen yield over 0.15g of Pt-loaded CdS (λg 420nm)from an aqueous solution containing 0.25M Na 2SO 3and 0.35M Na 2S under visible-light irradiation,four runs in one continuous reaction.(b)Influence of wavelength on the apparent quantum yield for hydrogen production over 0.15g of Pt-loaded CdS from fresh aqueous solution containing 0.25M Na 2SO 3and 0.35M Na 2S under visible-light irradiation with various wave-lengths of 420,470,500,520,560,and 600nm.114Chem.Mater.,Vol.20,No.1,2008Bao et al.the oxidation state of sacrificial regents suppressing the proceeding of photocatalytic reaction.Optimized photocatalytic conditions for the highest hy-drogen yield have been investigated,considering the factors such as the amount of photocatalyst,the content of loaded Pt nanocrystals,and the concentration of sacrificial regents.Figure 6a shows the influence of the amount of photocatalysts on the rate of hydrogen evolution.The rate of hydrogen evolution rapidly increases from 0.85mmol h -1in the presence of 0.05g of Pt-loaded CdS to the maximum rate of 4.1mmol h -1in the presence of 0.15g of Pt-loaded CdS and then decreases to 0.75mmol h -1in the presence of 0.45g of Pt-loaded CdS.The hydrogen production rate stably increases as the photons are continuously injected into the semiconductors.The usage ratio of all the photons from the irradiation in photocatalytic reaction containing diluted photocatalysts is low because a lot of photons either pass through the solution without being absorbed by the photo-catalysts or are reflected by the glass reactor wall.With increasing the concentration of photocatalysts,the number of photons absorbed by the photocatalysts increases and the number of photons reflected by the glass reactor decreases.The total amount of the photons absorbed by the photocata-lysts thus increases.As the concentration of photocatalysts reached a critical value at which the photons absorbed by the photocatalysts reached the stable maximum,however,the photons cannot be continuously injected into photocata-lyst particles,which increases the recombination of electrons and holes.Therefore,the hydrogen production rate decreases again.All in all,the reason is due to the combination effectof thermodynamic and kinetic factors of the photoelectron generation and transfer in heterogeneous photocatalytic reactions.An optimal amount of photocatalysts is required in order to attain the highest photocatalytic efficiency.In the present study,the optimal amount of photocatalysts is smaller than those of previous reported data because the present CdS nanostructures have relatively fine particle size,nanoporous structure,and much larger surface area.32,33Both bulk and surface electron/hole pairs quickly separate and migrate to the CdS -electrolyte interface via a much shorter distance,reducing water to hydrogen and oxidizing the sacrificial reagents.Figure 6b shows the dependence of the rate of hydrogen evolution upon the amount of Pt cocatalyst loaded on the CdS photocatalyst.The photocatalytic activity of the pure CdS photocatalyst was as low as 0.02mmol h -1and unstable but was greatly improved after loading with Pt nanocrystals.As shown in Figure 6b,the hydrogen yield rapidly increases from 0.55mmol h -1at a 0.5%Pt loading to the maximum rate of 4.1mmol h -1at a 13%Pt loading.However,with continuously increasing the Pt loading content,the hydrogen yield quickly decreases to 1.4mmol h -1at 15%Pt loading and then slowly decreases to 0.62mmol h -1at 40%Pt loading,which is due to a shielding effect of the Pt cocatalyst.We also observed that the as-made Pt-loaded CdS photo-catalyst becomes much darker with increasing the Pt-loading concentration,which is because of the increasing surface Pt coverage.The presence of metal particles,particularly Pt,on the surface of the CdS microcrystals drastically decreases the luminescence intensity.This expected quenching action of Pt indicates that Pt particles are very efficient traps for electrons.56Figure 6c shows the influence of the concentration of sacrificial regents of Na 2SO 3and Na 2S.H 2O is reduced to hydrogen by the electrons photogenerated in the conduction band accompanied by oxidation of sacrificial reagents.There exists an optimal concentration of 0.35M for Na 2SO 3and 0.25M for Na 2S for the maximal hydrogen evolution rate.Generally,concentrated sacrificial reagents could be expected for the better diffusion of the reacting species to the surface of photoatalysts.On the other hand,as the pH increases under basic conditions,the negative shift has been observed for both the flat-band potential of CdS and the redox potential of H +/H 2.However,the flat-band potential of CdS becomes less negative than the redox potential of H +/H 2,leading to a decrease in the reactivity of the CdS photocatalyst in the present of sacrificial regents,such as SO 32-,formic acid,formaldehyde,methanol,etc.57,58Further,the flat-band potential of CdS also depends on the concentration of S 2–.34With increasing the concentration of S 2-,the flat-band potential of CdS located more negative than the redox potential of H +/H 2.However,the S 22-ions,forming from the oxidation of S 2-ions,have a less negative reduction potential than photons,which results in a decreasing ef-(56)Hengleln,A.;Llndlg,B.;Westerhausen,J.J.Phys.Chem.1981,85,1627.(57)Matsumura,M.;Hiramoto,M.;Irhara,T.;Tsubomura,H.J.Phys.Chem.1984,88,248.(58)Matsumura,M.;Saho,Y.;Tsubomura,H.J.Phys.Chem.1984,88,3807.Figure 6.(a)Relation of the amount of CdS catalyst loaded with 10wt %Pt nanocrystals to the hydrogen evolution rate from an aqueous solution containing 0.25M Na 2SO 3and 0.35M Na 2S.(b)Relation of the Pt-loading content to the hydrogen evolution rate from an aqueous solution containing 0.25M Na 2SO 3,0.35M Na 2S,and 0.15g of CdS catalyst.(c)Relation of the concentration of sacrificial regents to the hydrogen evolution rate from an aqueous solution containing 0.15g of Pt-loaded CdS and sacrificial regents of Na 2S and Na 2SO 3at different concentrations.(d)Hydrogen evolution rates from aqueous solutions containing 0.25M Na 2SO 3,0.35M Na 2S,and 0.15g of 10wt %Pt-loaded CdS catalyst for each of the six 12h reations.All reactions were performed under visible-light irradiation provided by a light source,300W Xe lamp with a cutoff filter (λ>420nm).115Chem.Mater.,Vol.20,No.1,2008Self-Templated Synthesis of Nanoporous CdS。
肺粘液表皮样癌1例报告
肺粘液表皮样癌1例报告肺粘液表皮样癌(pulmonary mucoepidermoid caicinoma PMEC)是一种起源于气管、支气管粘膜下腺体Kulchitsky细胞的罕见的肺部恶性肿瘤,占所有肺癌的0.1-0.2%[1]。
PMEC的发病率无明显性别差异,发病年龄4-78岁不等, 成年人多见, 平均为28.5岁[2]。
在此,收集了 1例罕见的肺粘液表皮样癌的病例,现报告如下:临床资料患者,男性,51岁,在我院行胸部CT发现右肺下叶见大小约6*4cm肿块,增强后轻度不均匀强化,右肺下叶支气管狭窄伴阻塞。
癌胚抗原:52ng/ml,结合各项检查,考虑肿块恶性的可能性大,故在全麻下行单孔胸腔镜下肺癌根治术+纵膈淋巴结清除术。
术后病理:肿物大小4*2*3cm,切面呈灰白色,质硬伴大量粘液;右肺浸润性粘液腺癌,伴坏死,淋巴结未见转移。
免疫组化:CK7(+),TTF-1(+),NapsinA(+),p63(+),Ki-67(+)。
明确诊断:浸润性粘液表皮样癌T2bN0M0 ⅡA期高分化型。
讨论PMEC是一种源于支气管的恶性肿瘤,主要由分泌黏液细胞,表皮样细胞和未特殊分化中间细胞组成[3],病理上分为高分化黏液样癌和低分化黏液样癌,Ki-67指数可作为高分化和低分化的鉴别指标[4],指数越高,低分化的可能性越大,恶性的可能性越高。
高分化黏液表皮样癌是以黏液细胞为主,细胞异型性不明显,肿瘤多无坏死区域,可见钙化,行手术切除,预后良好;低分化黏液表皮样癌较少见,以中间细胞为主,异型细胞核分裂像和坏死表现及区域淋巴结转移为特征,可远处转移,手术切除,预后不佳。
免疫组化:P63、Ki-67, NapsinA、CK7和TTF-1及FISH检测MAML2可以识别PMEC,其中普遍认为t(11;19)(q21;p13)染色体易位形成的MAML2与粘液表皮样癌的发生相关,且具有高度特异性[10-11],因此成为了PMEC研究方向,并用于MAML2阳性的患者的靶向治疗。
ch1
1Overview and Historical PerspectiveNatural gas hydrates are crystalline solids composed of water and gas.The gas molecules(guests)are trapped in water cavities(host)that are composed of hydrogen-bonded water molecules.Typical natural gas molecules include methane,ethane,propane,and carbon dioxide.Historically,the research efforts on natural gas hydrates can be classified into three landmark phases that cover the following periods:•Thefirst period,from their discovery in1810until the present,includes gas hydrates as a scientific curiosity in which gas and water aretransformed into a solid.•The second period,continuing from1934until the present,predomin-antly concerns man-made gas hydrates as a hindrance to the natural gasindustry.•The third period,from the mid-1960s until the present,began with the discovery that nature predated man’s fabrication of hydrates by millionsof years,in situ in both the deep oceans and permafrost regions as wellas in extraterrestrial environments.As a result,the present is a culmination of three periods,representing the most fascinating and productive time in the history of natural gas hydrates.During thefirst century after their discovery,the number of hydrate publications totaled approximately40;in modern times,the number of hydrate publications,both in the technical and in the popular press,has increased dramatically with over400 publications in2005alone.The semilogarithmic plot of Figure1.1illustrates the exponential growth in the number of hydrate-related publications in the twenti-eth century.Table1.1lists reviews,chapters,and monographs on the subject of hydrates.The purpose of this chapter is to review the three periods mentioned above, as an overview and historical perspective.The major concepts will be discussed briefly;detailed investigations are presented in the following chapters.1.1H YDRATES AS A L ABORATORY C URIOSITYIn1778,Joseph Priestley performed cold experiments in his Birmingham labor-atory by leaving the window open before departing on winter evenings,returning the next morning to observe the result.He observed that vitriolic air(SO2)would impregnate water and cause it to freeze and refreeze,whereas marine acid air(HCl)12Clathrate Hydrates of Natural Gases10,0001000L o g a r i t h m s c a l e o f p u b l i c a t i o n s 100101212Decades of the twentieth century3456789104514379517246112973010F IGURE 1.1The growth of hydrate-related publications in the twentieth century by decade.(Reproduced from Sloan,E.D.,Am.Mineral.,89,1155(2004).With permission from the Mineralogy Society of America.)and fluor acid air (SiF 4)would not.With such experiments,Makogon and Gordejev (1992,unpublished data)suggest that Priestley might have discovered clathrate hydrates more than 30years before Davy’s discovery of clathrate hydrates.“It is water impregnated with vitriolic acid air that may be converted into ice,whereas water impregnated with fluor acid will not freeze ....I had observed that with respect to marine acid air and alkaline air (NH 3)that they dissolve ice,and that water impregnated with them is incapable of freezing,at least in such a degrees of cold as I had exposed them to.The same I find,is the case with fluor acid air,but it is not so at all with vitriolic acid air,which,entirely contrary to my expectation,I find to be altogether difficult ....But whereas water impregnated with fixed air discharges it when it is converted into ice,water impregnated with vitriolic acid air,and then frozen retains it as strongly as ever.”However,unlike Davy’s experiments,Priestley’s temperature (17◦F)of the gas mixture was below the ice point,so there is no unequivocal evidence that the frozen system was hydrate.There is also no record of validation experiments by Priestley;consequently,Davy’s independent discovery of chlorine hydrate is generally credited as the first observance.Natural gas hydrates were first documented by Sir Humphrey Davy (1811),with these brief comments on chlorine (then called oxymuriatic gas)in the Bakerian lecture to the Royal Society in 1810.“It is generally stated in chemical books,that oxymuriatic gas is capable of being condensed and crystallized at low temperature;I have found by several experiments that this is not the case.The solution of oxymuriatic gas in water freezes moreOverview and Historical Perspective3 T ABLE1.1Reviews,Chapters,and Monographs on Clathrate Hydrates1927Schroeder:Die Geschichte der Gas Hydrate1946Deaton and Frost:Gas Hydrates and Relation to the Operation of Natural-Gas Pipelines 1959Katz et al.:“Water–Hydrocarbon Systems”in Handbook of Natural Gas Engineering1967Jeffrey and McMullan:“The Clathrate Hydrates”in Progress in Inorganic Chemistry1973Davidson:“Clathrate Hydrates”in Water:A Comprehensive Treatise V ol.21974Makogan:Hydrates of Natural Gas1977Berecz and Balla-Achs:Gas Hydrates1980Kvenvolden and McMenamin:Hydrates of Natural Gas:A Review of Their Geologic Occurrence1983Cox,ed.:Natural Gas Hydrates:Properties,Occurrence and Recovery1983Lewin&Associates and Consultants:Handbook of Gas Hydrate Properties and Occurrence 1987Krason and Ciesnik:Geological Evolution and Analysis of Confirmed or Suspected Gas Hydrate Localities(13volumes)1988Holder et al.:“Phase Behavior in Systems Containing Clathrate Hydrates”Rev.Chem.Eng. 1990Katz and Lee:“Gas Hydrates and Their Prevention”in Natural Gas Engineering: Production and Storage1990Sloan:Clathrate Hydrates of Natural Gases1993Englezos:“Clathrate Hydrates”Ind.Eng.Chem.Res.1994Sloan,Happel and Hnatow,eds.:International Conference on Natural Gas Hydrates,NY 1995Kvenvolden,K.A.:A Review of the Geochemistry of Methane in Natural Gas Hydrate 1996Monfort,ed.:Second International Conference on Natural Gas Hydrates,Toulouse1997Makogon:Hydrates of Hydrocarbons1998Henriet and Mienert,eds.:Gas Hydrates:Relevance to World Margin Stability and Climate Change1998Ginsburg and Soloviev:Submarine Gas Hydrates2000Holder and Bishnoi,eds.:Third International Conference on Natural Gas Hydrates,Salt Lake City2000Sloan:Hydrate Engineering2000Paull et al.:Proc.Ocean Drilling Program,Science Results for Leg164(Blake Ridge) 2001Paull and Dillon,eds.:Natural Gas Hydrates:Occurrence,Distribution and Detection 2002Mori,ed.:Fourth International Conference on Natural Gas Hydrates,Yokohama2003Kennett et al.:Methane Hydrates in Quaternary Climate Change:The Clathrate Gun Hypothesis2003Max,ed.Natural Gas Hydrates in Oceanic and Permafrost Environments2004Taylor and Kwan,eds.:Advances in the Study of Gas Hydrates2004Zhang and Lanoil,eds.:“Geomicrobiology and Biogeochemistry of Gas Hydrates and of Hydrocarbon Seeps”in Chemical Geology2005Austvik,ed.:Fifth International Conference on Natural Gas Hydrates,Trondheim2005Dallimore et al.,eds.:“Report of the Mallik5L International Field Experiment on Recovering In Situ Hydrates from Permafrost”,Geological Survey of Canada Report. 2005IODP:Preliminary Report Leg311(Northern Cascadia Margin)2006Johnson et al.,eds.:Economic Geology of Natural Gas Hydrates2006Tréhu et al.:Ocean Drilling Program Scientific Report Leg2044Clathrate Hydrates of Natural Gases readily than pure water,but the pure gas dried by muriate of lime undergoes no change whatever at a temperature of40below0◦of Fahrenheit.”Over the following one and one-quarter centuries,researchers in thefield had two major goals,namely,(1)to identify all the compounds that formed hydrates and (2)to quantitatively describe the compounds by their compositions and physical properties.Table1.2provides a summary of the research over this period.T ABLE1.2Hydrates from1810to1934Year Event1810Chlorine hydrate discovery by Sir Humphrey Davy1823Corroboration by Faraday—formula Cl2·10H2O1882,1883Ditte and Mauménédisputed the composition of chlorine hydrates1884Roozeboom confirmed the composition as Cl2·10H2O1884LeChatelier showed that the Cl hydrate P–T curve changes slope at273K1828Bromine hydrates discovered by Löwig1876Br2hydrates corroborated by Alexeyeff as(Br2·10H2O)1829SO2hydrates found by de la Rive as SO2·7H2O1848Pierre determined the formula of SO2·11H2O1855Schoenfield measured the formula as SO2·14H2O1884,1885Roozeboom postulated upper/lower hydrate quadruple points using SO2as evidence;determined univariant dependence of P on T1856–1858CS2hydrate composition disputed by Berthelot(1856),Millon(1860),Duclaux (1867),Tanret(1878)1877,1882Cailletet and Cailletet and Bordetfirst measured mixed gas hydrates from CO2+PH3 and from H2S+PH31882de Forcrand suggested H2S·(12–16)H2O and measured30binary hydrates of H2S with a second component such as CHCl3,CH3Cl,C2H5Cl,C2H5Br,C2H3Cl.Heindicated all compositions as G·2H2S·23H2O1883Wroblewski measured carbon dioxide hydrates1885Chancel and Parmentier determined chloroform hydrates1888Villard obtained the temperature dependence of H2S hydrates1888de Forcrand and Villard independently measured the temperature dependence of CH3Cl hydrate1888Villard measured hydrates of CH4,C2H6,C2H4,C2H2,N2O1890Villard measured hydrates of C3H8and suggested that the temperature of the lower quadruple point is decreased by increasing the molecular mass of a guest;Villardsuggested hydrates were regular crystals1896Villard measured hydrates of Ar,and proposed that N2and O2form hydrates;first to use heat of formation data to get the water/gas ratio1897deForcrand and Thomas sought double(w/H2S or H2Se)hydrates;found mixed(other than H2S x)hydrates of numerous halohydrocarbons mixed with C2H2,CO2,C2H6 1902de Forcrandfirst used Clausius–Clapeyron relation for H and compositions;tabulated15hydrate conditions1919Scheffer and Meyer refined Clausius–Clapeyron technique1923,1925de Forcrand measured hydrates of krypton and xenonOverview and Historical Perspective5 In Table1.2,the following pattern was often repeated:(1)the discovery of a new hydrate was published by an investigator;(2)a second researcher disputed the composition proposed by the original investigator;and(3)a third(or more) investigator(s)refined the measurements made by the initial two investigators, and proposed slight extensions.As a typical example,in the case of chlorine hydrate after Davy’s discovery in1810,Faraday confirmed the hydrate(1823)but proposed that there were ten water molecules per molecule of chlorine.Then Ditte (1882),Mauméné(1883),and Roozeboom(1884)re-examined the ratio of water to chlorine.The period from1810to1900is characterized by efforts of direct composition measurements with inorganic hydrate formers,especially bromine,inorganics con-taining sulfur,chlorine,and phosphorus,and carbon dioxide.Other notable work listed in Table1.2was done by Cailletet and Bordet(1882),whofirst measured hydrates with mixtures of two components.Cailletet(1877)was also thefirst to measure a decrease in gas pressure when hydrates were formed in a closed cham-ber,using a precursor of an apparatus still in use at the Technical University of Delft,the Netherlands.1.1.1Hydrates of Hydrocarbons Distinguished fromInorganic Hydrates and IceTwo French workers,Villard and de Forcrand,were the most prolific researchers of the period before1934,with over four decades each of heroic effort.Villard (1888)first determined the existence of methane,ethane,and propane hydrates. de Forcrand(1902)tabulated equilibrium temperatures at1atm for15components, including those of natural gas,with the exception of iso-butane,first measured by von Stackelberg and Müller(1954).The early period of hydrate research is marked by a tendency to set an integral number of water molecules per guest molecule,due to the existing knowledge of inorganic stoichiometric hydrates that differed substantially from clathrate hydrates.For example,Villard’s Rule(1895)states that“all crystallize regularly and have the same constitution that can be expressed by the formula M+6H2O.”Schroeder(1927)noted that Villard’s Rule was followed by15of the17known gas hydrate formers.Today,we know that too many exceptions are required for Villard’s Rule to be a useful heuristic.Molecules approximated by Villard’s Rule are small guests that occupy both cavities of structures I or II(see Chapter2).It gradually became clear that the clathrate hydrates distinguished themselves by being both nonstoichiometric and crystalline;at the same time,they differed from normal hexagonal ice because they had no effect on polarized light.1.1.2Methods to Determine the Hydrate CompositionThe work in Table1.2illustrates one of the early research difficulties that is still present—namely,the direct measurement of the water to gas ratio in hydrates (hydration number,n=water molecules per guest).Whereas many solids6Clathrate Hydrates of Natural Gases such as carbon dioxide precipitate in a relatively pure form,or a form offixed composition,gas hydrate composition is variable with temperature,pressure,and the composition of associatedfluid phases.Although the composition measure-ment of either the gas or the water phase is tractable(usually via chromatography), measurement of the hydrate composition is more challenging.On a macroscopic basis,it is difficult to remove all excess water from the hydrate mass;this causes a substantial decrease in the accuracy of hydrate com-position measurements.Hydrate formations often occlude water within the solid in a metastable configuration,thereby invalidating the composition obtained upon dissociation.Mixed guest compositions of the hydrate are also confounded by the concentration of heavy components in the hydrate phase.Unless the associated gas reservoir is large,preferential hydration may result in variable gas consumption and perhaps an inhomogeneous hydrate phase as discussed in Chapter6.Villard(1896)proposed an indirect macroscopic method to determine hydra-tion number,which uses the heat of formation,both above and below the ice point. In his review,Schroeder(1927)indicates that after1900,researchers abandoned direct measurement of hydrate phase composition,preferring Villard’s method (see Section4.6.2)that relies on easier measurements of pressure and temperature. Miller and Strong(1946)provided another thermodynamic method to determine hydration number,discussed in Section4.6.2.2.Circone et al.(2005)obtained hydration numbers from direct macroscopic measurements of the amount of gas released during dissociation.Their results were in close agreement with those obtained by Galloway et al.(1970)from measurements of gas uptake during synthesis and release during decomposition, and by Handa(1986e)from calorimetric measurements.The advent of modern microscopic measurement tools and a means for bridging the microscopic and macroscopic domains(statistical thermodynamics)enable the direct determination of hydrate phase properties.The hydration number can be determined from single crystal or powder(using Rietveld refinement)x-ray and neutron diffraction.The hydration number can also be determined using Raman (Sum et al.,1997;Uchida et al.,1999)and NMR(Ripmeester and Ratcliffe, 1988)spectroscopy combined with statistical thermodynamics.Davidson et al. (1983)and Ripmeester and Ratcliffe(1988)first used NMR spectroscopy and Sum et al.(1997)first used Raman spectroscopy to determine the guest occupancies of each type of cage.Single crystal and powder x-ray and neutron diffraction (Udachin et al.,2002;Rawn et al.,2003)have also been applied to determ-ine guest occupancies and hydrate composition.These methods are discussed in Chapter6.1.1.3Phase Diagrams Provide Hydrate ClassificationRoozeboom(1884,1885)generated thefirst pressure–temperature plot for SO2 hydrate,similar to that in Figure1.2for several components of natural gases.In thefigure,H is used to denote hydrates,I for ice,V for vapor,and L w and L HC for aqueous and hydrocarbon liquid phases,respectively.For each component,Overview and Historical Perspective 78060402010864210.80.60.40.20.1268273278283288Temperature (K)P r e s s u r e (M P a )293298303I–H–V L W –H –V M e t h a n e L W –H –V E t h a n e L W –V –L H C L W –V –L H C H –L H C –V L W –V –L H C Q 1Q 1Q 1Q 1Q 2Q 2Q 2I –L W –H L W –H –L H C L W –H –L H C I –L W –H I –L W –V i-Butane L W –H –VH –V –L H C I –L W –V L W –H –V P r o p a n e I–H–V I–H–V I–H–V F IGURE 1.2Phase diagrams for some simple natural gas hydrocarbons that form hydrates.Q 1:lower quadruple point;Q 2:upper quadruple point.(Modified from Katz,D.L.,Cor-nell,D.,Kobayashi,R.,Poettmann,F.H.,Vary,J.A.,Elenbaas,J.R.,Weinaug,C.F.,The Handbook of Natural Gas Engineering ,McGraw Hill Bk.Co.(1959).With permission.)the hydrate region is to the left of the three phase lines (I–H–V),(L w –H–V),(L w –H–L HC );to the right,phases exist for liquid water or ice and the guest component as vapor or liquid.In Figure 1.2,the intersection of the above three phase lines defines both a lower hydrate quadruple point Q 1(I–L W –H–V)and an upper quadruple point Q 2(L W –H–V–L HC ).These quadruple points are unique for each hydrate former,providing a quantitative classification for hydrate components of natural gas.Each quadruple point occurs at the intersection of four three-phase lines (Figure 1.2).The lower quadruple point is marked by the transition of L W to I,so that with decreasing temperature,Q 1denotes where hydrate formation ceases from vapor and liquid water,and where hydrate formation occurs from vapor and ice.Early researchers took Q 2(approximately the point of intersection of line L W –H–V with the vapor pressure of the hydrate guest)to represent an upper temperature limit for hydrate formation from that component.Since the vapor pressure at the critical temperature can be too low to allow such an intersection,some natural gas components such as methane and nitrogen have no upper quadruple point,Q 2,and8Clathrate Hydrates of Natural Gases2CH 4CH 4 ? H 2OConcentrationT e m p e r a t u r e F IGURE 1.3Proposed CH 4–H 2O T –x phase diagram with the solid solution range (P ≈5MPa).Regions expanded for ease of viewing.(Reproduced from Huo,Z.,Hester,K.E.,Sloan,E.D.,Miller,K.T.,AIChE.J.,49,1300(2003).With permission.)consequently they have no upper temperature limit for hydrate formation.Phase diagrams are discussed in detail in Chapter 4.The isobaric methane–water phase diagram was produced by Kobayashi and Katz in 1949(Figure 1.3).This classical phase diagram represents the hydrate composition as a vertical constant composition line.This assumes that the hydrate is stoichiometric and that cage occupancy is independent of temperature or system composition.Reassessment of this phase diagram was initiated by the authors’laboratory in 2002(Huo et al.,2002,2003).We revisited the largely overlooked work by Glew and Rath (1966).Glew and Rath (1966)found from density meas-urements of sI ethylene oxide that nonstoichiometry (with the minimum occupancy of the small cages varying from 19%to 40%)can occur depending on the solution composition.This work validated the earlier statistical thermodynamic calcula-tions showing nonstoichiometry in clathrate hydrates (van der Waals and Platteeuw,1959).X-ray diffraction and Raman studies were performed to re-evaluate the rela-tion between hydrate and overall composition (Huo et al.,2002,2003).A modified methane–water phase diagram was proposed to include a small solid solution range of around 3%(Figure 1.3).[A solid solution is a solid-state solution of one or more solutes (guests)in a solvent (host framework).Generally,the crystal structure (of the clathrate hydrate)remains homogeneous and unchanged when substituting/Overview and Historical Perspective9 adding solutes(varying guest occupancies)to the solvent(host framework).] The solid solution range is represented by a parabolic hydrate region(attributed to incompletefilling of small cages of sI hydrate)in the isobaric methane–water phase diagram,which replaces the vertical stoichiometric hydrate line of Kobayashi and Katz(1949).1.2H YDRATES IN THE N ATURAL G AS I NDUSTRYIn the mid-1930s Hammerschmidt studied the1927hydrate review of Schroeder (D.L.Katz,Personal Communication,November14,1983)to determine that natural gas hydrates were blocking gas transmission lines,frequently at temperat-ures above the ice point.This discovery was pivotal in causing a more pragmatic interest in gas hydrates and shortly thereafter led to the regulation of the water content in natural gas pipelines.The detection of hydrates in pipelines is a milestone marking both the import-ance of hydrates to industry and the beginning of the modern research era.As a complement to the history prior to1934in Table1.2,hydrate studies in more recent times are indicated in Table1.3.The key scientific developments and applications to the natural gas industry are listed in Table1.3.With this listing as an abstract, an introduction to modern research is provided in the next few pages,with more details and literature references in later chapters.1.2.1Initial Experiments on Natural Gas HydratesAfter Hammerschmidt’s initial discovery,the American Gas Association commissioned a thorough study of hydrates at the U.S.Bureau of Mines.In an effort spanning World War II,Deaton and Frost(1946)experimentally investig-ated the formation of hydrates from pure components of methane,ethane,and propane,as well as their mixtures with heavier components in both simulated and real natural gases.Predictive method results are still compared to the Deaton and Frost data. It should be remembered,however,that while this study was both painstaking and at the state-of-the-art,the data were of somewhat limited accuracy,particu-larly the measurements of gas composition.As will be seen in Chapters4and5, small inaccuracies in gas composition can dramatically affect hydrate formation temperatures and pressures.For example,Deaton and Frost were unable to dis-tinguish between normal butane and iso-butane using a Podbielniak distillation column,and so used the sum of the two component mole fractions.Accurate composition measurement techniques such as chromatography did not come into common usage until early in the1960s.Many workers including Hammerschmidt(1939),Deaton and Frost(1946), Bond and Russell(1949),Kobayashi et al.(1951),and Woolfolk(1952) investigated the effects of inhibitors on hydrates.In particular,many chloride salts such as those of calcium,sodium,and potassium,were considered along with methanol and monoethylene glycol.Methanol gradually became one of the10Clathrate Hydrates of Natural Gases T ABLE1.3Milestones in Hydrate Studies since19341934Hammerschmidt discovers hydrates as pipeline plugs;provides Hammerschmidt equation;discovers thermodynamic inhibitors1941Katz et al.begin K-values and gas gravity methods to predict hydrate mixtures1946Deaton and Frost present data summary on hydrates and their prevention1949von Stackelberg reports20years of diffraction data on hydrate crystals1949Kobayashi begins a50year hydrate research effort with study of binary systems1951Claussen proposes,and von Stackelberg and Müller confirm sII unit crystal1952Claussen and Polglase,Müller and von Stackelberg,and Pauling and Marsh confirm sI unit crystal1954von Stackelberg and Jahn measure sII hydrate formed from two sI guest molecules1959van der Waals and Platteeuw(vdWP)propose statistical theory based on structure1960Robinson begins30year hydrate research effort with study of paraffin/olefin hydrates 1963McKoy and Sinanoglu apply Kihara potential to vdWP theory1963Davidson makesfirst dielectric measurements1965Kobayashi and coworkers apply vdWP theory to mixtures1966Davidson makesfirst broadline NMR measurements of hydrates1972Parrish and Prausnitz apply vdWP theory to natural gases1975Sloan begins measurements of two-phase hydrate equilibria1976Ng begins with three-and four-phase study of liquid hydrocarbons1976Holder et al.begin work with study of sI and sII coexistence and hydrates in earth1979Bishnoi and coworkers begin kinetic study with simulations of well blowouts1980Ripmeester and Davidson makefirst pulsed NMR measurements1982Tse and coworkers begin molecular dynamic(MD)simulation of hydrates1984Davidson et al.confirm Holder’s suggestion that small,simple guests form sII1984Handa begins study of calorimetry and phase equilibria1985John and Holder determine effect of higher order coordination shells in vdWP theory 1986Englezos begins study of kinetics of methane,ethane dissociation1987Ripmeester and coworkers discover new structure H(sH)hydrates1988Danesh,Todd,and coworkers begin four phase experiments with hydrates1990a,b Rodger studies relative stability using MD simulation1991Mehta obtains sH data,applied vdWP theory to CH4+large(>8Å)guest(s)1991Behar et al.introduce water emulsification concept to control hydrate blockage1991Sloan proposes molecular mechanism with kinetic inhibition implications1992Kotkoskie et al.show that hydrates are controlled by drilling mud water activity1996Sum measures hydrate composition and hydration number using Raman spectroscopy 1997Kuhs et al.publishfirst report of double occupancy of nitrogen molecules in large cage of sII hydrate at high pressures,exceeding several hundred bar1997Udachin et al.reportfirst single crystal x-ray diffraction measurements of a sH gas hydrate1997Dyadin et al.discover a very high pressure phase of methane hydrate that is stable up to 600MPa1999Dyadin et al.discover that H2forms a clathrate hydrate at high pressures up to1.5GPa 2004Camargo et al.and BP/SINTEF introduce“coldflow”concept to prevent hydrate plug formation without the need of chemical additivesmost popular inhibitors,due to its ability to concentrate in free water traps after being vaporized into the upstream gas.Effects of thermodynamic inhibitors such as methanol are quantified in Chapters 4, 5, 6, and 8.1.2.2Initial Correlation of Hydrate Phase EquilibriaWhen Hammerschmidt(1934)identified hydrates in pipelines,he published a cor-relation summary of over100hydrate formation data points.Shortly afterward, Professor D.L.Katz and his students at the University of Michigan began an exper-imental study.Because it was impractical to measure hydrate formation conditions for every gas composition,Katz determined two correlative methods.The initial predictive method by Wilcox et al.(1941)was based on distribution coefficients(sometimes called K vsi values)for hydrates on a water-free basis.With a substantial degree of intuition,Katz determined that hydrates were solid solutions that might be treated similar to an ideal liquid solution.Establishment of the K vsi value(defined as the component mole fraction ratio in the gas to the hydrate phase) for each of a number of components enabled the user to determine the pressure and temperature of hydrate formation from mixtures.These K vsi value charts were generated in advance of the determination of hydrate crystal structure.The method is discussed in detail in Section4.2.2.The second(and simplest available)method,generated by Professor Katz (1945)and students in a graduate class,is presented in Figure1.4.The plot enabled the user to estimate a hydrate formation pressure,given a temperature and gas gravity(gas molecular weight divided by that of air).The original work also enabled the determination of the hydrate formation limits due to expansions of natural gases,as in throttling gas through a valve.This method and its limitations are discussed in detail in Section4.2.1as a usefulfirst approximation for hydrate formation conditions.Katz’s two predictive techniques provided industry with acceptable pre-dictions of mixture hydrate formation conditions,without the need for costly measurements. Subsequently, hydrate research centered on the determination of the hydrate crystal structure(s).Further refinements of the K vsi values were determined by Katz and coworkers (especially Kobayashi) in Chapter 5of the Handbook of Natural Gas Engineering(1959),by Robinson and coworkers (Jhaveri and Robinson,1965;Robinson and Ng,1976),and by Poettmann(1984).1.2.3Hydrate Crystal Structures and Hydrate T ypeDefinitionsIn the late1940s and early1950s,von Stackelberg and coworkers summarized two decades of x-ray hydrate crystal diffraction experiments at the University of Bonn.The interpretation of these early diffraction experiments by von Stackelberg (1949,1954,1956),von Stackelberg and Müller(1951a,b),Claussen(1951a,b), and Pauling and Marsh(1952)led to the determination of two hydrate crystal structures(sI and sII)shown in Figure1.5.。
荧光原位杂交技术在临床实验室应用的现存问题和对策
荧光原位杂交技术在临床实验室应用的现存问题和对策李江超;朱颖慧;陈宝江;关新元【摘要】FISH的技术在临床的应用日渐广泛,成为快速诊断和确诊官颈癌、乳腺癌、白血病等疾病的一项分子诊断技术.但是,到目前为止国家相关部门尚未出台相应的实验室条件要求、人员培训以及标准的操作流程,同时该技术实验操作和结果判断等环节较多,对临床结果影响较大,鉴于此,我们就目前临床实验中尚未解决和需要关注的地方进行阐述,以便更好地改进和规范实验技术步骤,有效减少临床检验的误诊.【期刊名称】《分子诊断与治疗杂志》【年(卷),期】2011(003)002【总页数】4页(P111-114)【关键词】FISH;实验室标准;操作标准化;实验结果判读【作者】李江超;朱颖慧;陈宝江;关新元【作者单位】中山大学肿瘤防治中心实验研究部,广东,广州,510060;中山大学肿瘤防治中心实验研究部,广东,广州,510060;中山大学附属第一医院产前诊断中心,广东,广州,510080;中山大学肿瘤防治中心实验研究部,广东,广州,510060【正文语种】中文1.1 FISH技术及其原理荧光原位杂交技术(Florescence in situ hybridization, FISH)是1986年产生的分子生物学和细胞遗传学结合的技术[1]。
其原理是用已知的荧光直接标记核酸探针,按照碱基互补的原则,与待检材料中未知的单链核酸进行异性结合,形成可被检测的杂交双链核酸。
由于DNA分子在染色体上是沿着染色体纵轴呈线性排列,因而可以探针直接与染色体进行杂交从而将特定的基因在染色体上定位。
在荧光显微镜下观察信号的强弱,从而诊断疾病或者寻找基因定位。
1.2 临床应用目前该技术已经由以往的科研逐步转向临床,在美国,某些FISH检测项目已获FDA批准,如HER-2基因检测对乳腺癌的诊断和治疗的应用[2]。
目前,中国卫生主管部门也批准了5个FISH检测项目,其中就包括HER-2[2]。
多色荧光原位杂交技术在尿脱落细胞学阴性膀胱癌患者术后随访监测中的应用
多色荧光原位杂交技术在尿脱落细胞学阴性膀胱癌患者术后随访监测中的应用王南雄;王尉;吕军;王健;叶纯【摘要】目的探讨多色荧光原位杂交技术( M-FISH)在尿脱落细胞学阴性膀胱癌患者术后随访监测中的应用价值. 方法收集接受经尿道膀胱肿瘤切除术后的膀胱尿路上皮癌患者76例,术前、术后随访尿脱落细胞学检查均阴性,分别采用膀胱镜、M-FISH进行术后随访监测.结果 76例患者中14例(18.4%)复发,M-FISH仅检出3例(3.9%)复发,漏诊11例(14.5%). 膀胱镜检查结果阴性54例,复发1例(低级别),M-FISH检查结果阳性. 膀胱镜检查结果可疑12例,复发4例(低级别2例、高级别2例) ,M-FISH检查结果阳性1例(低级别). 膀胱镜检查结果阳性10例,复发9例(低级别5例、高级别4例) ,M-FISH检查结果阳性1例(高级别). 膀胱镜检查结果阴性、M-FISH检查结果为阳性预测复发的敏感性为100%、特异性94 .3%,阳性预测值25%,阴性预测值100%. 膀胱镜检查结果可疑、M-FISH检查结果为阳性预测复发的敏感性为25%、特异性87.5%,阳性预测值50%,阴性预测值70%. 膀胱镜、M-FISH检查结果均为阳性预测复发的敏感性为11.1%、特异性100%,阳性预测值100%,阴性预测值11 .1%. 结论 M-FISH检测在尿脱落细胞学阴性膀胱癌患者术后随访复发中的检出率不高,经济效益低,不考虑作为膀胱癌术后随访的常规监测项目.%Objective To evaluate the application value of multicolor fluorescence in situ hybridization ( M-FISH) in postoperative follow-up monitoring of bladder cancer patients with negative urinecytology .Methods A total of 78 patients with urothelial cancer who were treated by transurethral resection of the bladder were included in this study .All of them had negative urine cytology .The cystoscopy and M-FISHwere used to monitor the recurrence of the disease , and the results were compared.Results In 76 patients, 14 cases recurred (18.4%).M-FISH only detected 3 cases of recurrence (4%), and 11 cases of missed diagnosis (14.5%).Cystoscopy showed 54 cases with negative results , 1 case of recur-rence (low level), and the M-FISH showed positive results.Cystoscopy showed 12 cases were suspicious and 4 cases of re-currence (2 cases of low level, 2 cases of high level), and M-FISH showed 1 case with positive results (low level).Cysto-scopy showed 10 cases with positive results , 9 cases of recurrence ( 5 cases of low level , 4 cases of high level ) , and M-FISH showed 1 case with positive results ( high level ) .For that cystoscopy showed negative results , M-FISH showed posi-tive results, the sensitivity of predicting recurrence was 100%, the specificity was 94.3%, the positive predictive value was 25%, and the negative predictive value was 100%.For that cystoscopy showed suspicious results , M-FISH showed positive results, the sensitivity of predicting recurrence was 25%, the specificity was 87.5%, the positive predictive value was 50%, and the negative predictive value was 70%.For that cystoscopy showed positive results , M-FISH showed posi-tive results, the sensitivity of predicting recurrence was 11.1%, the specificity was 100%, the positive predictive value was 100%, and the negative predictive value was 11.1%.Conclusions M-FISH assay in postoperative follow-up monito-ring of bladder cancer patients with negative urine cytology had a low detection rate and low economic benefits .It was not considered as a follow-up routine monitoring project in bladder cancer surveillance after transurethral resection .【期刊名称】《山东医药》【年(卷),期】2015(055)017【总页数】3页(P7-9)【关键词】膀胱肿瘤;多色荧光原位杂交技术;尿脱落细胞学【作者】王南雄;王尉;吕军;王健;叶纯【作者单位】公安边防总医院,广东深圳518000;广州军区广州总医院;广州军区广州总医院;公安边防总医院,广东深圳518000;公安边防总医院,广东深圳518000【正文语种】中文【中图分类】R737.25膀胱尿路上皮癌占膀胱癌90%以上,其发病率和病死率呈逐年上升趋势[1]。
聚苯胺类复合材料在生物医学领域的研究进展
聚苯胺类复合材料在生物医学领域的研究进展窦成福;张玉梅【期刊名称】《宁夏医科大学学报》【年(卷),期】2013(035)003【总页数】4页(P353-355,封4)【关键词】聚苯胺;生物传感器;组织工程;药物释放;生物相容性【作者】窦成福;张玉梅【作者单位】宁夏医科大学基础医学院,银川,750004【正文语种】中文【中图分类】O631导电聚合物又称导电高分子,是指通过掺杂等手段将电导率控制在半导体和导体之间的聚合物。
在众多导电聚合物中,聚苯胺(polyaniline,PANi)具有许多优异性能如导电性、氧化还原性、催化性能、电致变色行为、质子交换性质及光电性质等,而且原料易得、合成简便,易于工业化,因此,具有一定的实际应用价值。
聚苯胺是由苯胺单体经化学氧化或电化学氧化聚合而成的。
目前,聚苯胺被广泛应用于气体传感器[1]、超级电容器[2]、抗腐蚀[3]、电池[4]等领域。
从所从事的专业角度出发,本文主要总结了近年来聚苯胺在生物医学领域中的研究发展情况。
1 生物传感器生物传感器是聚苯胺在生物医学领域研究最多的一个方面,聚苯胺在构建仿生界面方面具有诸多优点,表现为易于在电极表面成膜;膜的厚度、生物组分的固定量易于控制,制备的传感器重现性好;通过改变高聚物的氧化还原态,可调节酶的生物反应活性;易于微型化和多功能化。
科研工作者以聚苯胺为载体材料制作了多种传感器,如酶传感器、DNA传感器、免疫传感器等,实现了对葡萄糖[5]、胆固醇[6]、过氧化氢[7]、NADH[8]、病毒基因[9]、炭疽热[10]等多达几十种生物组分的痕量检测。
除了可进行单一组分检测外,利用聚苯胺还可构建双酶传感器进行多组分的同时检测。
如Yang等[11]采用原位合成法制备出金@聚苯胺核壳型纳米复合材料,该复合材料在中性甚至碱性溶液中仍然具有很好的电活性,用其制备的电化学传感器可同时检测多巴胺和抗坏血酸。
与此同时,纳米科技的发展为生物传感器的发展开辟了新的方向。
DH3初筛阳性在宫颈高危病变中的临床意义
DH3初筛阳性在宫颈高危病变中的临床意义何婷婷;阮萍;彭忠异;姚平;刘莹【摘要】目的:探讨DH3(人乳头瘤病毒杂交捕获—化学发光法核酸检测试剂盒)初筛阳性在宫颈高危病变中的临床意义.方法:回顾性统计我院2017年6月—2018年8月门诊或体检行DH3筛查的患者,其中175例DH3阳性的患者作为研究对象,以病理活检为金标准,统计DH3阳性结果的病变分布情况.结合TCT的检查结果,探讨细胞学在DH3阳性结果的分流作用,同时运用统计学分析年龄、病毒负荷量与宫颈高危病变的关系.结果:DH3阳性率15.95%,病理学示175例DH3阳性患者中,炎症63例(36.00%),LSIL42例(24.00%),HSIL44例(25.14%),宫颈癌(鳞癌及腺癌)26例(14.86%),宫颈高危病变(≥HSIL)率40.00%,DH3阳性预测值64.00%;宫颈高危病变率在年龄组20~30岁、30~40岁、40~50岁、50~60岁、>60岁中分别为12.50%、25.58%、45.61%、53.13%、68.42%,宫颈高危病变随年龄增加而增高,组间差异有统计学意义(χ2=22.863,P<0.05);宫颈高危病变率在HPV负荷量分组≤10、10~30、30~60、60~100、>100中分别为14.89%,28.57%、47.62%58.35%、55.41%,宫颈高危病变随高危病毒感染量增加而增高,组间差异有统计学意义(χ2=22.026,P<0.05);157例DH3阳性患者同时行TCT及病理活检,TCT阳性病变(≥ASC-US)83例,阳性率52.87%,组织学阳性病变(≥LSIL)97例,阳性率61.78%,TCT与组织学一致率为74.52%(117/157).结论:DH3是宫颈高危病变筛查的可行方法,随着年龄及感染病毒负荷量的增加,宫颈高危病变发生率增加,TCT对DH3阳性患者分流有重要指导意义,但同时也需关注患者年龄及病毒负荷量对宫颈高危病变的影响.【期刊名称】《医学理论与实践》【年(卷),期】2019(032)006【总页数】4页(P803-805,813)【关键词】人乳头瘤病毒;宫颈高危病变;筛查;分流;TCT【作者】何婷婷;阮萍;彭忠异;姚平;刘莹【作者单位】广西中医药大学附属瑞康医院病理科,广西南宁市 530011;广西中医药大学附属瑞康医院病理科,广西南宁市 530011;广西中医药大学附属瑞康医院病理科,广西南宁市 530011;广西中医药大学附属瑞康医院病理科,广西南宁市530011;广西中医药大学附属瑞康医院病理科,广西南宁市 530011【正文语种】中文【中图分类】R711.74宫颈癌是女性常见的恶性肿瘤,其发生发展与人乳头瘤状病毒(HPV)感染密切相关,特别是高危型HPV(Highrisk human papillomavirus,HR-HPV)的持续感染[1]。
- 1、下载文档前请自行甄别文档内容的完整性,平台不提供额外的编辑、内容补充、找答案等附加服务。
- 2、"仅部分预览"的文档,不可在线预览部分如存在完整性等问题,可反馈申请退款(可完整预览的文档不适用该条件!)。
- 3、如文档侵犯您的权益,请联系客服反馈,我们会尽快为您处理(人工客服工作时间:9:00-18:30)。
Department of Statistics and O.R., University of Granada, Spain Available online 26 September 2007
∗ Corresponding author. Departamento de Estadística e I.O., Facultad de Ciencias, Universidad de Granada, Campus Fuentenueva s/n. 18071-Granada, Spain. Tel.: +34 95824630-mail addresses: aaguiler@ugr.es (A.M. Aguilera), escabias@ugr.es (M. Escabias), valderra@ugr.es (M.J. Valderrama).
Computational Statistics & Data Analysis 52 (2008) 3187 – 3197 /locate/csda
Forecasting binary longitudinal data by a functional PC-ARIMA model
0167-9473/$ - see front matter © 2007 Elsevier B.V. All rights reserved. doi:10.1016/j.csda.2007.09.015
3188
A.M. Aguilera et al. / Computational Statistics & Data Analysis 52 (2008) 3187 – 3197
ANOVA to model a scalar response variable from a functional covariate, and functional canonical correlation analysis to investigate different modes of variability in two sets of curves. An excellent review of functional methods and their applications in a diverse range of subject areas has been developed (Ramsay and Silverman, 2005, 2002). The actual development on statistical methods for analyzing functional data and new trends have been recently revised (González-Manteiga and View, 2007). In the general context of FDA, logistic regression has been recently extended to model a time-independent binary response in terms of a functional predictor (continuous time stochastic process). This model has been used to predict the probability of a high risk birth outcome from periodically stimulated foetal heart rate tracings (Ratcliffe et al., 2002). Usually, principal component (PC) regression and principal covariate regression are used to forecast a response variable from highly correlated predictors (see Heij et al., 2007 for a comparison of the forecast accuracy of these two methods). Different FPCA approaches for estimating the functional logit model have been proposed (Escabias et al., 2004). In the general context of functional generalized linear models, the EM algorithm has been used for estimating the model (James, 2002). An odds ratio interpretation of the relationship between a binary response and a functional predictor in terms of the estimated parameter function of the functional logit model has been established and an application with climatological data has been developed (Escabias et al., 2005). A PLS approach for estimating the functional logit model has been recently introduced (Escabias et al., 2007). With the same objective, a linear discriminant analysis for classification of functional data has been proposed (Preda et al., 2007). The estimation of this model is based on functional PLS regression (Preda and Saporta, 2005). Alternative nonparametric curves discrimination methods have been studied (Ferraty and Vieu, 2003). FPCA is a generalization of the classic principal component analysis (PCA) of a sample of data vectors for the reduction of dimension of a set of sample curves (Ramsay and Silverman, 2005). Different models based on FPCA to forecast a functional predictor in the future from its past evolution have been developed in recent years. Principal component prediction (PCP) models based on linear regression of each future PC on a reduced set of past PCs were introduced (Aguilera et al., 1997). PCP models have been adapted for predicting a continuous time series from unequally spaced discrete time observations (Aguilera et al., 1999a, b). A detailed study of PCP models can be found in Valderrama et al. (1997). Mixed FPC-ARIMA models are based on ARIMA modelling of the PCs of the sample functions obtained by cutting the observed time series in periods of the same amplitude (Valderrama et al., 2002). All these FPCA models allow not only to forecast a continuous time series in a whole future interval but also to reconstruct it between the discretization time points in the past. This mixed approach has been recently extended allowing application to a broader class of problems such as robust forecasting of mortality and fertility rates (Hyndman and Ullah, 2007). On the other hand, a time-localized frequency domain PCA method is proposed for signals that exhibit locally stationary behavior (Ombaoa and Hob, 2006). A functional logit model based on mixed FPC-ARIMA modelling of the functional predictor, which allows us to forecast the time evolution of a binary response from discrete time observations of a continuous time series, is introduced in this paper. In Section 2, we formulate the problem in mathematical terms. A functional logistic regression (FLR) model for predicting binary longitudinal data, in terms of the functional predictor sample curves obtained by cutting the original time series in periods of the same amplitude, is proposed in Section 3. In order to solve the multicollinearity problem and to reduce dimension, FLR is estimated in Section 4 by using as covariates a reduced set of functional PCs of the functional predictor. ARIMA modelling of each PC series will be considered in Section 5 to forecast the predictor continuous time series in a future period of time followed by the forecast of the related binary response in this period. Finally, a climatological application with real data will be developed in Section 6. We aim to predict the risk of drought in a future period of time from monthly observations of El Niño phenomenon. 2. Problem formulation Let us suppose that we have observations of a continuous time series {x(t)} at discrete time points in the interval (0, NT] and one observation Yw of a related binary response Y at each period of amplitude T defined as ((w − 1)T , wT ], w = 1, . . . , N. Let us denote by twk (w = 1, . . . , N ; k = 1, . . . , mw ) the observation time points of the original time series {x(t)} at each period ((w − 1)T , wT ] and by xwk = x(twk ) the corresponding observed values. Thus, the purpose is to estimate a functional logit model to forecast the binary response in future periods ((w∗ − 1)T , w∗ T ](w ∗ > N), from the forecasting of the series x(t) provided by a mixed ARIMA-FPCA model in such periods.