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RECSIT1.1中英文对照全文Assessment of the change in tumour burden is an important feature of the clinical evaluation of cancer therapeutics: both tumour shrinkage (objective response) and disease progression are useful endpoints in clinical trials. Since RECIST was published in 2000, many investigators, cooperative groups, industry and government authorities have adopted these criteria in the assessment of treatment outcomes. However, a number of questions and issues have arisen which have led to the development of a revised RECIST guideline (version 1.1). Evidence for changes, summarised in separate papers in this special issue, has come from assessment of a large data warehouse (6500 patients), simulation studies and literature reviews.临床上评价肿瘤治疗效果最重要的一点就是对肿瘤负荷变化的评估：瘤体皱缩（目标疗效）和病情恶化在临床试验中都是有意义的判断终点。

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Recent development of in silico molecular modeling for gas and

Available online at Recent development of in silico molecular modeling for gas and liquid separations in metal–organic frameworksJianwen JiangAs a new family of nanoporous materials,metal–organic frameworks(MOFs)are considered versatile materials for widespread applications.Majority of current studies in MOFs have been experimentally based,thus little fundamental guidance exists for the judicious screening and design of task-speciﬁc MOFs.With synergistic advances in mathematical methods,computational hardware and software,in silico molecular modeling has become an indispensable tool to unravel microscopic properties in MOFs that are otherwise experimentally inaccessible or difﬁcult to obtain.In this article,the recent development of molecular modeling is critically highlighted for gas and liquid separations in MOFs.Bottom-up strategies have been proposed for gas separation in MOFs,particularly CO2capture.Meanwhile, interest for liquid separation in MOFs is growing and modeling is expected to provide in-depth mechanistic understanding. Despite considerable achievements,substantial challenges and new opportunities are foreseeable in more practical modeling endeavors for economically viable separationsin MOFs.AddressDepartment of Chemical and Biomolecular Engineering,National University of Singapore,117576,SingaporeCorresponding author:Jiang,Jianwen(chejj@.sg)Current Opinion in Chemical Engineering2012,1:138–144This review comes from a themed issue onNanotechnologyEdited by Hua Chun ZengAvailable online23rd December20112211-3398/$–see front matter#2011Elsevier Ltd.All rights reserved.DOI10.1016/j.coche.2011.11.002IntroductionDuring the past decade,metal–organic frameworks (MOFs)have emerged as a new family of nanoporous materials[1,2].In remarkable contrast to traditional inor-ganic zeolites,MOFs can be synthesized from various inorganic clusters and organic linkers,thus possess a wide range of surface area and pore size.More fascinatingly, the judicious selection of building blocks allows the pore volume and functionality to be tailored in a rational manner.With such salient features,MOFs are considered versatile materials for widespread potential applications [3,4]as illustrated in Figure1.Indeed,MOFs have been identiﬁed as a topical area in materials science and technology because of their implications for global and national economies[5].To date,thousands of MOFs have been synthesized in this vibrantﬁeld and several(Cu-BTC,ZIF-8,MIL-53,etc.) are commercially available under the trade name Basoli-te TM[6].However,massive research efforts on MOFs have been primarily based on experiments.It is impractical to search for task-speciﬁc MOFs by trial-and-error from inﬁnitely large number of possible candidates.Therefore, quantitative guidelines are desired for the high-throughput screening of enormous MOFs and the rational design of new MOFs towards practical applications.In this context, clear and deep microscopic understanding from a molecu-lar level is indispensable.With synergistic advances in mathematical methods,computational hardware and soft-ware,in silico molecular modeling has played an increas-ingly important role in unraveling microscopic properties in MOFs[7 ,8 ,9 ].Sophisticated modeling and simu-lation provide molecular insights that are experimentally intractable,if not impossible,thus elucidate underlying physics from bottom-up.Among many potential appli-cations of MOFs,separations are of central importance in chemical industry and have been actively investigated [10].In this article,the recent development of molecular modeling is critically highlighted for both gas and liquid separations in MOFs,and the foreseeable challenges and opportunities are discussed.Gas separationThe overwhelming majority of studies for gas separation in MOFs have been focused on CO2capture.This is because the combustion of fossil fuels produces a huge quantity of CO2emissions into the atmosphere.Carbon capture and sequestration is crucial to environmental protection and sustainable economy.As an essential pre-requisite,CO2has to be captured fromﬂue gas/ shifted syngas in post-/pre-combustion processes. Another important gas separation involving CO2is puri-ﬁcation of natural gas,in which impurities such as CO2 need to be separated to enhance calorie content.MOF adsorbentsMost synthesized MOFs are crystallites and tested as adsorbents for gas separation.Several reviews have sum-marized numerous experimental studies for CO2capture in MOF adsorbents[11–13].Nevertheless,nearly all these experiments examined the adsorption of pure gases (e.g.CO2,N2,CH4,and H2)due to the formidable difﬁculty associated with mixtures.By contrast,simu-lation can be readily used for single or multi-componentsystems.Thus,quantitative understanding of mixture adsorption in MOFs has been obtained,to a large extent,from simulation studies.Several bottom-up strategies as illustrated in Figure 2have been proposed to tune CO 2capture performance,for example,using speciﬁc MOFs with small pores,catenation,functionalization,ionic fra-meworks,exposed metals or metal doping.Yang and Zhong [14]simulated the adsorption of CO 2/CH 4/H 2mixture in two MOFs (IRMOF-1and Cu-BTC)and found pore size strongly affects separation efﬁciency.However,IRMOF-1and Cu-BTC do not possess iden-tical topology,leading to ambiguous interplay with the effect of pore size.In this regard,Babarao et al.[15]examined the separation of CO 2/CH 4mixture in isostruc-tural MOFs (Cu-BTC and PCN-60)and observed that the selectivity in Cu-BTC with small pores is nearly twice of that in PCN-60.This strategy of small pores is also reﬂected in framework catenation that can induce con-stricted pores and greater potential overlaps.For example,catenated IRMOF-13and PCN-6exhibit a larger selectivity for CO 2/CH 4mixture than non-cate-nated counterparts [15].An appealing strategy is to use ionic MOFs as demonstrated by Jiang and co-workers [16,17 ,18]for the separation of CO 2-containing mixtures.Simulation reveals that CO 2molecules are strongly adsorbed onto the ionic frameworks and nonframeworkions,and the predicted selectivity is signiﬁcantly higher than in neutral MOFs and many other nanoporous materials.On the other hand,Yazaydin et al.[19]screened a diverse set of 14MOFs for low-pressure CO 2capture from ﬂue gas combining simulation and experiment.The results show that M/DOBDC (M =Zn,Mg,Co or Ni)with high density of exposed metals strongly interact with CO 2.By physical and chemical doping,Xu et al.[20 ]estimated the separation of CO 2/CH 4mixtures in Li-modiﬁed MOF-5.Owing to the enhancement of electro-static potentials,adsorption selectivity was predicted to be much higher than in MOF-5.In a separate study,Lan et al.[21 ]simulated CO 2capture in covalent-organic frameworks doped by alkali,alkaline-earth and transition metals,and concluded that Li is the best surface modiﬁer for CO 2capture.The strategies outlined in Figure 2have been compre-hensively discussed [24 ,25 ].Two of them (ionic fra-meworks and metal doping)appear to be more efﬁcient to enhance CO 2capture.It should be noted that these strategies also can tune the separation of other mixtures,for example,the selectivity of alkane isomers was found to be enhanced by framework catenation [26].In a recent perspective,Krishna and van Baten [27 ]highlighted the potency of simulation in screening of best MOFs for CO 2capture and hydrocarbon separation,and they furtherRecent development of in silico molecular modeling Jiang 139Figure 1Purification Toxics RemovalDrug DeliveryFuel Cell SystemsStorageStorage and SeparationCarbon SequestrationSensingMOPWidespread potential applications of MOFs (/ees6/clathrates/index.shtml ).compared MOFs against traditional zeolites with regard to separation characteristics.As an alternative to simulation,analytical theories have been developed for gas separation in MOFs.Liu et al.[28,29]proposed a density functional theory (DFT)in 3D-nanoconﬁned space.The theory was applied to adsorption and separation in 3D-MOFs with complex pore networks,whereas most DFT studies are limited in simple conﬁned geometries (e.g.slit and cylindrical pores).Good agree-ment was obtained between theoretical predictions,simu-lation and experimental data.Coudert et al.[30]developed the osmotic framework adsorbed solution theory (OFAST)in terms of a competition between host’s free energy and adsorption energy.This theory is based exclusively on pure-component adsorption and has the superior capability to describe ﬂexible MOFs.For illustration,the authors used the OFAST to examine the effect of breathing on separation of CO 2/CH 4mixtures in MIL-53.The modeling studies discussed above for gas separation in MOF adsorbents are primarily focused on adsorption selectivity.However,several other factors (e.g.working capacity,regenerability,etc.)should be included in prac-tice as discussed by Bae and Snurr [31 ].Another crucial issue is how moisture in gas mixtures would affect sep-aration performance?From systematical simulation stu-dies in various neutral and ionic MOFs,Jiang and coworkers observed four different intriguing effects ofH 2O on CO 2capture [25 ].It is also instructive to examine structural change in ﬂexible MOFs that might occur upon adsorption [32].The incorporation of ﬂexi-bility to simulate structural change would need a robust force ﬁeld.However,a general force ﬁeld is currently unavailable for MOFs and ﬁrst-principles modeling is expected to play a pivotal role [33 ].In addition,the chemical and thermal stability of MOFs are important for separation [34].A large number of MOFs are unstable in atmosphere or under moisture,which impedes their util-ization.Therefore,it is indispensable to develop molecu-lar guidelines for the design of stable MOFs.Nevertheless,unraveling what govern the stability of MOFs at a microscopic level is a challenge.MOF membranesCompared with adsorptive separation,membrane-based separation is considered to be energetically more efﬁ-cient,lower capital cost and larger separation capability.However,the fabrication of MOF membranes is a for-midable task [35].Only in recent years,have there been active experimental endeavors to explore MOF mem-branes for gas separation [36 ].Since both equilibrium and dynamic properties are required,simulation for gas separation in MOF mem-branes is more time-consuming than in MOF adsorbents.Nevertheless,a handful of simulation studies have been reported.Keskin and Sholl [37 ]examined the separation140NanotechnologyFigure 2functionalizationmetal dopingionic frameworksexposed metalssmall porescatenationBottom-up strategies to tune CO 2capture performance.The representative MOFs are from [15,17 ,20 ,22,23].performance of diverse MOFs for CO2/CH4and CO2/H2 mixtures.They found that all the MOFs examined exhi-bit unfavorably low CO2selectivities and mixture effects play a crucial role in determining the performance.By combining simulation and IR microscopy,Bux et al.[38] simulated ethene/ethane separation in ZIF-8membrane. They found that ethane adsorbs more strongly than ethene,but ethene diffuses faster;and the interplay results in a membrane permeation selectivity for ethene. Krishna and van Baten[27 ]underlined the advantages of using simulation tools in the screening of MOF mem-branes for CO2capture.Along with considerable interest in MOF membranes, MOF-based composite membranes have received increasing attention for gas separation.In this emerging area,a handful of experiments have been conducted[39], but modeling studies are ing atomistic simulation and continuum model,Keskin and Sholl[40]attempted to select MOF/polymer membranes for high-perform-ance gas separation.A highly selective MOF was ident-iﬁed and predicted to enhance the performance of Matrimid and other polymers for CO2/CH4separation. Chen et al.[41]proposed a composite with ionic liquid (IL)supported on IRMOF-1.The simulation reveals that ions in the composite act as favorable sites for CO2adsorption,and the selectivity for CO2/N2mixture is higher than in neat IL,IRMOF-1and many other supported IL membranes.It is worthwhile to note that defects and inter-crystalline interstices usually exist in synthesized MOF membranes. Nevertheless,most simulation studies use perfect and rigid models for MOF membranes.How to incorporate defects and interstices into practical modeling is challenging.On the other hand,theﬂexibility of MOF structures may have a larger inﬂuence in membrane separation than adsorbent separation[36 ],and should be implemented as well into modeling.Another essential issue is the mech-anical properties of MOFs[42].The high pressure exerted for membrane separation may distort MOF structures and deteriorate performance.It is thus crucial to quantitatively understand how pressure affects pore geometries and framework dimensionalities.For MOF-based composite membranes,microscopic insights into the interactions between MOF and other species(e.g.polymer or ionic liquid)are strikingly important and fundamental studies at a molecular level are desired.Liquid separationWhile gas separation in MOFs has been extensively investigated,endeavors for liquid separation are lagged behind[43 ].A recent trend has been to explore the use of MOF adsorbents and membranes for liquid separation. By combining chromatographic and breakthrough exper-iments,Alaerts et al.determined the adsorption and separation of ortho-substituted alkylaromatics(xylenes, ethylbenzene,ethyltoluenes and cymenes)in a column packed with MIL-53crystallites[44].Jin and coworkers tested the separation of water/organics mixtures in MIL-53membrane and observed a high selectivity for water removal from ethyl acetate solution[45].Simulation for liquid separation in MOFs is scarce owing to the signiﬁcant amount of computational time required to sample liquid phase.Consequently,the microscopic understanding of liquid separation in MOFs is far from complete.To the best of our knowledge,only two simu-lation studies have been reported in this area,one for water desalination and the other for biofuel puriﬁcation. Recent development of in silico molecular modeling Jiang141Figure3Selectivities of biofuel in Na-rho-ZMOF and Zn4O(bdc)(bpz)2by pervaporation[47 ].Speciﬁcally,Hu et al.[46]performed simulation on the desalination of NaCl aqueous solution through a ZIF-8 membrane by reverse osmosis.Because of the sieving effect of small apertures in ZIF-8,Na+and ClÀions could not transport through ZIF-8membrane and water desa-lination was observed.Theﬂux of water permeating the membrane was found to scale linearly with external pressure.In a separate study,Nalaparaju et al.[47 ] examined hydrophilic Na-rho-ZMOF and hydrophobic Zn4O(bdc)(bpz)2for biofuel puriﬁcation.The selectiv-ities between water and ethanol in the two MOFs are largely determined by adsorption behavior.As indicated in Figure3,Na-rho-ZMOF is preferable to remove water, whereas Zn4O(bdc)(bpz)2is promising to enrich ethanol. The simulation provides molecular guidelines for the selection of appropriate MOFs towards efﬁcient biofuel puriﬁcation.Currently,modeling for liquid separation in MOFs is very limited.With increasing demands for clean water,liquid fuels and other liquid-based applications,more efforts are expected in order to provide deep molecular insights.A pre-requisite for liquid separation is that the MOFs used should be stable in water or other liquids[48],it is crucial to understand what factors govern the stability of MOFs, which would allow to produce stable MOFs for liquid separation.ConclusionAs a burgeoningﬁeld,research activities in MOFs are rather hectic.In addition to enormous experimental stu-dies,we have witnessed the recent development of in silico molecular modeling for MOFs.Microscopic under-standing has been achieved for gas separation particularly CO2capture in MOFs,and bottom-up strategies have been proposed to enhance separation efﬁciency.How-ever,liquid separation in MOFs remains largely unex-plored at a molecular level and more endeavors are desired towards this end.It is obvious that current molecular modeling for separ-ations using MOFs is still in an infant stage.As discussed above,substantial challenges are foreseen for more prac-tical modeling and precise description.A number of issues should be considered in future modeling,such as the stability and mechanical properties of MOFs, structuralﬂexibility,material regenerability,and effect of moisture(in gas separation).These challenges provide new opportunities for modeling studies to unravel in-depth microscopic insights and thus provide quantitative guidelines on the rational screening and design of novel MOFs.Furthermore,for energy-efﬁcient and cost-effec-tive separations,process requirements are essential to be integrated with material properties at a system level.In this context,molecular modeling,process optimization, as well as material synthesis,should be synergized holi-stically towards the development of best MOFs for economically viable separations and other practical appli-cations.AcknowledgementsThe author gratefully acknowledges the National University of Singapore, the Singapore National Research Foundation,and the Ministry of Education of Singapore for support.References and recommended readingPapers of particular interest,published within the period of review, have been highlighted as:of special interestof outstanding interest1.Yaghi OM,O’Keefe M,Ockwig NW,Chae HK,Eddaoudi M,Kim J:Reticular synthesis and design of new materials.Nature2003, 423:705-714.2.Long JR,Yaghi OM:The pervasive chemistry of metal–organicframeworks.Chem Soc Rev2009,38:1213-1214.3.MacGillivray LR(Ed):Metal–Organic Frameworks:Design andApplication.Hoboken,New Jersey:John Wiley&Sons,Inc.;2010.4.Farrusseng D(Ed):Metal–Organic Frameworks:Applications fromCatalysis to Gas Storage.Weinheim,Germany:Wiley-VCH;2011.5.Adams J,Pendlebury D:Global Research Report:MaterialsScience and Technology.Leeds,UK:Thomson Reuters;2011. 6.Czaja AU,Trukhan N,Muller U:Industrial applications of metal–organic frameworks.Chem Soc Rev2009,38:1284-1293.7.Keskin S,Liu J,Rankin RB,Johnson JK,Sholl DS:Progress,opportunities,and challenges for applying atomically detailed modeling to molecular adsorption and transport in metal–organic framework materials.Ind Eng Chem Res2009,48:2355-2371.This article reviews the applications of atomically detailed modeling for MOFs.Quantum mechanical calculations are used to examine structural and electronic properties,while molecular modeling calculations are used to study adsorption and diffusion in MOFs.8.DU¨ren T,Bae YS,Snurr RQ:Using molecular simulation tocharacterise metal–organic frameworks for adsorptionapplications.Chem Soc Rev2009,38:1203-1212.This review gives an overview of how molecular simulation can be used to characterize MOFs for gas adsorption,and how molecular insights can be combined to develop design principles for speciﬁc applications.9.Jiang JW,Babarao R,Hu ZQ:Molecular simulations for energy, environmental and pharmaceutical applications of nanoporous materials:from zeolites,metal–organic frameworks to protein crystals.Chem Soc Rev2011,40:3599-3612.This review summarizes the recent simulation studies for energy,envir-onmental and pharmaceutical applications of nanoporous materials ran-ging from zeolites,MOFs to protein crystals with increasing degree of complexity in building blocks.Major challenges in future simulation exploration are discussed.10.Li JR,Sculley J,Zhou HC:Metal–organic frameworks forseparations.Chem Rev2012,112,doi:10.1021/cr200190s,inpress.11.Keskin S,van Heest TM,Sholl DS:Can metal–organicframework materials play a useful role in large-scale carbon dioxide separations?ChemSusChem2010,3:879-891.12.Fe´rey G,Serre C,Devic T,Maurin G,Jobic H,Llewellyn PL,Weireld GD,Vimont A,Daturi M,Chang JS:Why hybrid porous solids capture greenhouse gases?Chem Soc Rev2011,40:550-562.13.Li JR,Ma YG,McCarthy MC,Sculley J,Yu JM,Jeong HK,Balbuena PB,Zhou HC:Carbon dioxide capture-related gasadsorption and separation in metal–organic frameworks.Coord Chem Rev2011,255:1791-1823.14.Yang QY,Zhong CL:Molecular simulation of CO2/CH4/H2mixture adsorption in metal–organic frameworks.J Phys Chem B2006,110:17776-17783.142Nanotechnology15.Babarao R,Jiang JW,Sandler SI:Adsorptive separation of CO2/CH4mixture in metal–exposed,catenated and charged metal–organic frameworks:insight from molecular simulation.Langmuir2009,25:5239-5247.16.Jiang JW:Charged soc metal–organic framework for high-efﬁcacy H2adsorption and syngas puriﬁcation:atomisticsimulation study.AIChE J2009,55:2422-2432.17. Babarao R,Jiang JW:Unprecedentedly high selective adsorption of gas mixtures in rho zeolite-like metal–organic framework:a molecular simulation study.J Am Chem Soc 2009,131:11417-11425.A simulation study is reported for the separation of CO2-containingmixtures in rho zeolite-like MOF(ZMOF)with anionic framework.Fortheﬁrst time,this study characterizes nonframework Na+ions,examines gas separation in ionic ZMOF,and reveals that rho-ZMOF is a promisingcandidate for CO2capture.18.Babarao R,Eddaoudi M,Jiang JW:Highly porous ionic rhtmetal–organic framework for H2and CO2storage andngmuir2010,26:11196-11203.19.Yazaydin AO,Snurr RQ,Park TH,Koh K,Liu J,LeVan MD,Benin AI,Jakubczak P,Lanuza M,Galloway DB et al.:Screeningof MOFs for CO2capture fromﬂue gas using a combinedexperimental and modeling approach.J Am Chem Soc2009,131:18198-18199.20. Xu Q,Liu DH,Yang QY,Zhong CL,Mi JG:Li-modiﬁed metal–organic frameworks for CO2/CH4separation.J Mater Chem 2010,20:706-714.The separation of CO2/CH4mixtures is simulated in Li-modiﬁed MOF-5. The selectivity is found to be greatly improved owing to the enhancement of electrostatic potential by Li doping.21. Lan JH,Cao DP,Wang WC,Smit B:Doping of alkali,alkaline-earth,and transition metals in covalent-organic frameworks for enhancing CO2capture byﬁrst-principles calculations and molecular simulations.ACS Nano2010,4:4225-4237.This study examines the doping of a series of alkali(Li,Na,and K), alkaline-earth(Be,Mg,and Ca),and transition(Sc and Ti)metals in covalent-organic frameworks,and the effects of the doped metals on CO2capture.22.Dietzel PDC,Johnsen RE,Fjellvag H,Bordiga S,Groppo E,Chavan S,Blom R:Adsorption properties and structure of CO2 adsorbed on open coordination sites of metal–organicframework Ni2(dhtp)from gas adsorption.IR spectroscopyand X-ray diffraction.Chem Commun2008,44:5125-5127.23.Babarao R,Dai S,Jiang DE:Functionalizing porous aromaticframeworks with polar organic groups for high-capacity and selective CO2separation:a molecular simulation study.Langmuir2011,27:3451-3460.24. Liu DH,Zhong CL:Understanding gas separation in metal–organic frameworks.J Mater Chem2010,20:10308-10318.This feature article summarizes the recent advances of computer model-ing on gas separation in MOFs and demonstrates how computer model-ing can help to understand the separation characteristics of MOFs.Several strategies are proposed to improve the separation efﬁciency of MOFs.25. Jiang JW:Metal–organic frameworks for CO2capture:what are learned from molecular simulations.In Coordination Polymers and Metal Organic Frameworks.Edited by OO L,Ramı´rez LD.Nova Science Publishers;2011.In this book chapter,recent simulation studies are summarized for CO2 capture in MOFs.A number of strategies are discussed towards improv-ing capture performance.In addition,the effects of moisture in various MOFs on CO2adsorption and separation are presented.26.Babarao R,Tong YH,Jiang JW:Molecular insight intoadsorption and diffusion of alkane isomer mixtures in metal–organic frameworks.J Phys Chem B2009,113:9129-9136.27. Krishna R,van Baten JM:In silico screening of metal–organic frameworks in separation applications.Phys Chem Chem Phys 2011,13:10510-10593.This perspective highlights the potency of molecular simulation in deter-mining the best MOF for a given separation task.A variety of metrics that quantify separation performance such as adsorption selectivity,working capacity,diffusion selectivity and membrane permeability are determined by simulation.28.Liu Y,Liu HL,Hu Y,Jiang JW:Development of a densityfunctional theory in three-dimensional nanoconﬁned space:H2storage in metal–organic frameworks.J Phys Chem B2009, 113:12326-12331.29.Liu Y,Liu HL,Hu Y,Jiang JW:Density functional theory foradsorption of gas mixtures in metal–organic frameworks.J Phys Chem B2010,114:2820-2827.30.Coudert FX,Mellot-Draznieks C,Fuchs AH,Boutin A:Predictionof breathing and gate-opening transitions upon binary mixture adsorption in metal–organic frameworks.J Am Chem Soc2009,131:11329-11331.31.Bae YS,Snurr RQ:Development and evaluation of porousmaterials for carbon dioxide separation and capture.AngewChem Int Ed2011,50:11586-11596.The question of how a large number of MOFs can be quickly evaluated for CO2separation is addressed.Five adsorbent evaluation criteria are described and used to assess over40MOFs for their potential in CO2 separation processes for natural gas puriﬁcation,landﬁll gas separation, and CO2capture from power-plantﬂue gas.32.Horike S,Shimomura S,Kitagawa S:Soft porous crystals.NatChem2009,1:695-704.33.Taﬁpolsky M,Amirjalayer S,Schmid R:Atomistic theoreticalmodels for nanoporous hybrid materials.MicroporousMesoporous Mater2010,129:304-318.Available atomistic theoretical models are overviewed for the new class of functional porous hybrid materials such as MOFs and COFs.The current status of both periodic and non-periodic quantum mechanic,as well as molecular mechanic models are discussed.34.Kang IJ,Khan NA,Haque E,Jhung SH:Chemical and thermalstability of isotypic metal–organic frameworks.Chem Eur J2011,17:6437-6442.35.Shekhah O,Liu J,Fischer RA,Woll C:MOF thinﬁlms:existingand future applications.Chem Soc Rev2011,40:1081-1106. 36.Caro J:Are MOF membranes better in gas separation thanthose made of zeolites.Curr Opin Chem Eng2011,1:77-83. MOF membranes developed and tested for gas separation during the past5years have been reviewed.The structuralﬂexibility of MOFs prevents a sharp molecular sieving effect.Mixed-matrix membranes containing MOFs are predicted for the near future.37.Keskin S,Sholl DS:Efﬁcient methods for screening of metalorganic framework membranes for gas separations usingatomically detailed ngmuir2009,25:11786-11795. An efﬁcient approximate method is introduced to screen MOF mem-branes for gas separation with a connection between mixture adsorption and mixture self-diffusion properties.The method is applied to MOF membranes with chemical diversity for light gas separation.38.Bux H,Chmelik C,Krishna R,Caro J:Ethene/ethane separationby ZIF-8membrane:molecular correlation of permeation,adsorption,diffusion.J Membr Sci2011,369:284-289.39.Vinh-Thang H,Kaliaguine S:MOF-based mixed-matrix-membranes for industrial applications.In CoordinationPolymers and Metal Organic Frameworks.Edited by Ortiz OL,Ramı´rez LD.Nova Science Publishers;2011.40.Keskin S,Sholl DS:Selecting metal organic frameworks asenabling materials in mixed matrix membranes for highefﬁciency natural gas puriﬁcation.Energy Environ Sci2010,3:343-351.41.Chen YF,Hu ZQ,Gupta KM,Jiang JW:Ionic liquid/metal–organicframework composite for CO2capture:a computationalinvestigation.J Phys Chem C2011,115:21736-21742.42.Tan JC,Cheetham AK:Mechanical properties of hybridinorganic-organic framework materials:establishingfundamental structure–property relationships.Chem Soc Rev 2011,40:1059-1080.43.Cychosz KA,Ahmad R,Matzger AJ:Liquid phase separation by crystalline microporous coordination polymers.Chem Sci2010,1:293-302.This perspective details the experimental studies reported on liquid-phase separation using microporous coordination polymers(MCPs).Guest mole-cules examined include those as small as water to large organic dyes.In many cases,MCPs outperform zeolites and activated carbons in both kinetics and efﬁciency.Recent development of in silico molecular modeling Jiang143。

不同提取方法对井冈蜜柚皮精油组成与性质的影响

李欣，华建新，罗杰洪，等. 不同提取方法对井冈蜜柚皮精油组成与性质的影响[J]. 食品工业科技，2024，45（3）：83−97. doi:10.13386/j.issn1002-0306.2023030289LI Xin, HUA Jianxin, LUO Jiehong, et al. Effects of Different Extraction Methods on the Composition and Properties of Jinggang Pomelo Peel Essential Oil[J]. Science and Technology of Food Industry, 2024, 45(3): 83−97. (in Chinese with English abstract). doi:10.13386/j.issn1002-0306.2023030289· 研究与探讨 ·不同提取方法对井冈蜜柚皮精油组成与性质的影响李　欣1，华建新1，罗杰洪1，王国庆2，陈　赣2，周爱梅1,*（1.华南农业大学食品学院，广东省功能食品活性重点实验室，广东广州 510642；2.吉安井冈农业生物科技有限公司，江西吉安 343016）摘　要：以井冈蜜柚皮精油（Jinggang pomelo peel essential oil ，JPPEO ）为研究对象，采用水蒸气蒸馏法、低温连续相变法两种方法进行提取，以精油得率为主要指标，研究了萃取温度、压力、时间等因素对井冈蜜柚皮精油得率的影响，并通过正交法进行低温连续相变法提取工艺优化，同时对精油的理化性质及化学组成进行分析。

研究表明，低温连续相变提取井冈蜜柚皮精油（Low-temperature continuous phase transition extraction essential oil ，L-JPPEO ）的最佳工艺为：颗粒度30目，萃取温度55 ℃，萃取压力0.6 MPa ，萃取时间60 min ，解析温度70 ℃，此时精油得率为10.99‰，比水蒸气蒸馏法提取的精油（Hydro distillation essential oil ，H-JPPEO ）得率高出了2.88倍；理化性质实验结果表明，低温连续相变萃取的井冈蜜柚皮精油的不饱和脂肪酸含量较高，游离脂肪酸含量较低，酯类成分含量较低；傅里叶衰减全反射中红外光谱法（Fourier transform infrared spectroscopy ，FTIR ）鉴定出L-JPPEO 和H-JPPEO 含萜烯类化合物、醇类、酚类、醛类以及含羰基化合物。

阿斯利康、葛兰素史克、诺华、辉瑞四家公司大牛做临床前安全性评价经验

Decreasing the high attrition rate in the drug discovery and development process is a primary goal of the pharmaceutical industry. One of the main challenges in achieving this goal is striking an appropriate balance between drug efficacy and potential adverse effects 1as early as possible in order to reduce safety-related attrition, particularly in the more expensive late stages of clinical development. Gaining a better understanding of the safety profile of drug candidates early in the process is also crucial for reducing the likelihood of safety issues limiting the use of approved drugs, or even leading to their market withdrawal, bearing in mind the growing societal and regulatory emphasis on drug safety.It has been estimated that about 75% of all adverse drug reactions (ADRs) are dose-dependent and can be predicted on the basis of the pharmacology profiles of the candi-date compound (known as type A ADRs)2,3. The pharmacology profile of a preclinical compound can be classified into primary or secondary effects. Primary effects relate to the action of the compound at its intended target (or targets), whereas secondary effects are due to interactions with targets other than the primary target (or targets) (that is, off-target interactions). Off-target inter a ctions are often the cause of ADRs in animal models or clinical studies, and so careful characterization and identification of secondary pharmacology profiles of drug candidates early in the drug discovery process might help to reduce the incidence of type A ADRs.In vitro pharmacological profiling involves the screening of compounds against a broad range of targets (receptors, ion channels, enzymes and transporters) that are distinct from the intended therapeutic target (or targets) in order to identify specific molecular interactions that may cause ADRs in humans. New drugs pass through non-clinical safety pharmacology and preclinical toxicology assessments according to guidelines pro-duced by the International Conference on Harmonisation (ICH): for example, ICH S7A 4, ICH S7B 5 and ICH M3(R2)6. These guidelines describe the (mostly in vivo ) safety testing of drug candidates and are designed to prevent serious ADRs from occurring in clinical studies.The only in vitro pharmacology assay that is absolutely required by regulatory authorities is one that measures the effects of new chemical entities on the ionic current of native (I Kr ) or heterologously expressed human voltage-gated potassium channel subfamily H member 2 (KCNH2; also known as hERG)5. The mechanism bywhich blockade of hERG can elicit poten-tially fatal cardiac arrhythmias (torsadesde pointes) following a prolongation of theQT interval is well characterized 7,8, and theseriousness of this ADR is one reason why this assay is a mandatory regulatory require-ment. Receptor binding studies are also recommended as the first-tier approach for the assessment of the dependence potentialof novel chemical entities 9.However, current regulatory guidance does not describe which targets should constitute an in vitro pharmacological pro-filing panel and does not indicate the stage of the discovery process at which in vitro pharmacological profiling should occur.Nevertheless, the general trend for most pharmaceutical companies is to perform this testing early in drug discovery to reduce attrition and to facilitate better prediction of ADRs in the later stages of drug discovery and development.Here, for the first time, four major pharmaceutical companies (AstraZeneca, GlaxoSmithKline, Novartis and Pfizer) share their knowledge and experiences of the innovative application of existing screening technologies to detect off-target interactions of compounds. The objective of this article is to describe the rationale and main advan-tages for the use of in vitro pharmacological profiling, to discuss best practices and to share more widely the minimum panel of targets that, based on our collective experi-ence, should be considered. We hope this will enable smaller companies and academic institutions to benefit from this knowledge and consider joining us in our collaborative knowledge sharing. The impact of generating such data during the different phases of thedrug discovery process is illustrated throughcase studies.Reducing safety-related drug attrition: the use of in vitro pharmacological profilingJoanne Bowes, Andrew J. Brown, Jacques Hamon, Wolfgang Jarolimek, Arun Sridhar, Gareth Waldron and Steven Whitebread Abstract | In vitro pharmacological profiling is increasingly being used earlier inthe drug discovery process to identify undesirable off-target activity profiles that could hinder or halt the development of candidate drugs or even lead to market withdrawal if discovered after a drug is approved. Here, for the first time, the rationale, strategies and methodologies for in vitro pharmacological profiling at four major pharmaceutical companies (AstraZeneca, GlaxoSmithKline, Novartis and Pfizer) are presented and illustrated with examples of their impact on the drug discovery process. We hope that this will enable other companies and academic institutions to benefit from this knowledge and consider joining us in our collaborative knowledge sharing.PERSPECTIVES|Drug DiscoveryImpactAdvantages of in vitro profiling Early profiling of compounds against targets that are known to underlie ADRs (using the screening technologies devised for drug discovery and in particular for high-throughput screening) can help medicinal chemists to identify chemical series that lack this activity and are a potential liability, long before the final compound is selected (FIG. 1). Classical examples of well-characterized targets include hERG 7,8 (which is linked to cardiac arrhythmias, as noted above) and the 5-hydroxytryptamine (serotonin) receptor 2B (5-HT 2B ) (which is linked to cardiac valvulopathy)10,11. If inherently ‘clean’ chemical series (that is, those with no off-target activities of safety concern) are not available at the lead selection stage, an understanding of the structure–activity relationship (SAR) can help to reduce or eliminate off-target activity while retaining or increasing activity at the primary target.There is a growing awareness that in vitro pharmacological profiling 12,13, together with traditional safety pharmacology, can have a positive impact on the success rates of late-stage clinical development 14,15; BOX 1 summarizes the main advantages of in vitro pharmacological profiling. Although the impact of in vitro pharmacological profiling data differs according to the stage at which the data are utilized in the drug discovery process, it can be maximized when gener-ated throughout the discovery process (FIG. 1). Impact can be categorized into fiveaspects: first, early hazard identification and decision-making in the lead generation and selection phase; second, hazardelimination in the lead optimization phase; third, candidate selection; fourth, integrated risk assessment in early development; and fifth, risk management and mitigation in preclinical and clinical development.First, once a chemical series has been identified and activity at the primary thera-peutic target has been confirmed, secondary profiling can be performed to assess its promiscuity (that is, the percentage of targets hit at a specific concentration com-pared to the total number of targets tested). The importance of assessing promiscuity at this stage is discussed below. In addition, individual off-target activities at molecular targets with established linkage to ADRs can be identified at this stage.Second, these data can then be used to develop a lead optimization plan, which can include analysis of the SAR at off-targets in order to influence chemical design. An advantage of in vitro profiling at this stage means that testing only requires a relatively short turnaround time (days) compared to in vivo studies, which may take weeks to read out. The compound amounts required are also substantially lower for in vitro studies compared to in vivo studies.Third, at the end of lead optimization, the data can be used to select the candidate drug from a shortlist. The data can also be used to trigger and influence the design of in vivo discovery safety studies as part of an early integrated risk assessment. Most compounds are tested during the lead selection and lead optimization phases, as this is when the full benefits of chemical optimization in reducing potential liabilities can be achieved.Fourth, during the later phases of lead optimization, the value of the data is enhanced when interpreted in the context of the predicted therapeutic free plasma concentration , which is often available by this stage. Based on the drug’s affinity at the off-target and the predicted exposure levels, drug occupancy at off-targets can be esti-mated and safety margins can be calculated (this is described in more detail in the case study examples below).Fifth, in the preclinical and clinical development stages, when regulatory safety studies are performed, a broad understanding of the pharmacological profile of the candi-date drug is helpful for understanding the mechanistic underpinnings of the effects observed in vivo . In addition, the data can be used to build a patient risk management plan for Phase I trials if necessary. In some cases, additional end points or biomarkers may be included, which is a main advantage of profiling human targets from the begin-ning of the process.The overall benefits of performingin vitro pharmacological profiling through-out the drug discovery and development process are clear. However, so far, there is little consensus regarding the minimal panelFigure 1 | Alignment of in vitro pharmacology profiling to the drug discovery and development process. In the early phases of the drug discovery process, the main objective is to identify hazards and under-stand the potential for promiscuity (that is, having affinity at a broad range of targets) in the initial lead series. The application of a standard panel of targets (fewer than 60), annotated for linkage to safety liability, provides data that can be used for early decision-making regarding which lead series to select for further development. Data from this initial hazard identification profiling can be used in the lead optimization phase to eliminate the liability by building structure–activity relationship (SAR) models and using the data to drive chemical design. The profiling panelcan also be used to select key candidates to progress into developmentas well as to trigger and influence the design of investigative in vivo studies. In the later phases of drug discovery, data can be generated in a broader panel of targets and used for mechanistic understanding of effects in vivo . Importantly, it is desirable to obtain a comprehensive understanding of the broad pharmacological profile of the drug candidate before carrying out first-in-human trials.P E R S P E C T I V E Sof targets that should be used in these profiling campaigns. Below, we describe the pooling of information from four pharmaceutical companies, along with some of the lessons that have been learnt.‘Cross-pharma’ knowledgeIn vitro pharmacological profiling evolved in similar ways within different pharmaceutical companies. In the past, as part of the general pharmacology studies that were routinely carried out, a few advanced leads were tested against small panels of targets from protein classes closely related to the therapeutic target. With the development of screening technologies that enabled increased throughput at reduced costs, targets were added from members of major target classes that are involved in the regulation of vital systems such as the cardiovascular, respira-tory and nervous systems.The panels designed by these four phar-maceutical companies aim to cover a broad range of targets (receptors, ion channels, enzymes and transporters), and the inclu-sion of these targets is carefully considered by the weight of evidence for each target based on multiple parameters. These include knowledge of the following concepts:the expression and fundamental role of thetarget in physiology (including knockout mouse models and genetic evidence from humans); whether pharmacological modula-tion translates into a biological effect; and, more importantly, whether there is evidence from clinical (especially post-marketing) safety data that links an ADR to a specific target.Key to whether clinical experience links an ADR to a specific target is the testingof marketed drugs through in vitro assays. As a result of an improved understanding of the pharmacological significance of each target, but also owing to budgetary con-straints (see below for more details), most companies have reduced the number of targets that are included in their in vitro profiling panels. Now, only those targets that have a strong relevance to safety are routinely profiled. However, it should be noted that the application of larger, more diverse profiling panels on key drug candidates is also useful to facilitate a comprehensive understanding of their pharmacological profile before con-ducting first-in-human studies.One of the main factors affecting each company’s decision to include or exclude a particular target is the cost–benefit calcula-tion of the probability of a hit at the target compared to the magnitude of the impact of this hit. Targets with a high hit rate and a high impact (for example, hERG andmuscarinic acetylcholine receptor M1) arereadily prioritized for inclusion, whereastargets with a low hit rate and a low impactare readily prioritized for exclusion.More judgement is required for poten-tially high-impact targets that have a low hitrate. For example, the endothelin A receptoris included in the panel despite having a lowhit rate (<1%), as the cardiovascular andpotential teratogenic effects of ligands atthis receptor are profound and may not beidentified until long-term in vivo studies areperformed16. Conversely, melatonin receptorMT3has an extremely high hit rate (>30%)but as this has no definitive linkage to anadverse outcome, this target is not includedin the minimum panel. Targets that havesignificant potential impact but very low hitrates (for example, transient receptor poten-tial cation channel subfamily V member 4(TRPV4), which has hit rates of <0.5%) areoften not included based on this cost–benefitassessment. Each company has applied theseconsiderations independently to derive itstarget panel. Hit rates are assessed at 10 μMwith a broad range of chemically diversecompounds (more than 1,000 compounds)from each company’s compound library andare continuously monitored over periods ofroutine screening. In general, there was agood correlation among the four companiesregarding the hit rates at each target.The minimal panel of targetsHere, by comparing the lists of targets fromeach of the four companies, we have defineda minimum panel of targets that are testedin at least three out of the four companies.Despite large variations in the size of thepanels, as well as the tactics and technologiesused among the companies, there is sub-stantial overlap among the screened targets.Nineteen molecular targets were identicalin all four companies and another 25targets were tested in three out of the fourcompanies.The evolution of the target panels wasbased on decades of drug discovery experi-ence with the targets and their validationin the clinic, so it is understandable thatmost targets are from the G protein-coupledreceptor (GPCR) superfamily, with 24 rep-resentatives from 12 subfamilies. There areseven ion channel targets, six intracellularenzymes, three neurotransmitter transporters,two nuclear hormone receptors and onekinase. At present, we recommend that these44 targets are considered as a minimal panelthat should provide a broad early assessmentP E R S P E C T I V E Sof the potential hazard of a compound or chemical series. The targets, annotated with physiological relevance, are listed in TABLE 1.GPCRs. The GPCR targets include repre-sentatives from the major neurotransmitter classes (adenosine, adrenergic, cannabinoid, dopamine, histamine, muscarinic, opioid and serotonin receptors) as well as from peptidergic GPCRs (cholecystokinin, endothelin and vasopressin receptors).There was a good consensus on which GPCRs to include in the minimal panel; however, the decision on which muscarinic receptors to include in the panel provides an example of the impact of the individual experiences in each company. Muscarinic receptors have a fundamental role in physio-logy and their unintended inhibition or activation should therefore be avoided 17,18. All four companies test against the M 2 receptor subtype (owing to the well-known cardiovascular effects of M 2 receptor ago-nists and antagonists), and two test against M 1 and M 3 receptors in addition to M 2 receptors. The other two companies test either M 1 or M 3 receptors (in particular for the cognitive and gastric effects of M 1 receptor antagonists and the involvement of M 1 and M 3 receptors in constipation, disturbed vision and dry mouth). The ration-ale for only testing either M 1 or M 3 receptors (and not both) is that despite the undesir-ability of affecting either of these receptor subtypes, there is likely to be redundancy; that is, a compound that is not designed to have activity at a specific muscarinic receptor is likely to be non-selective across multiple members of the muscarinic class of receptors. This type of knowledge can be used to inform a risk–benefit analysis for the inclusion of only one of the receptor sub-types in the in vitro pharmacological panel rather than multiple members of a family. However, there is much flexibility with regard to deciding which receptor subtypes to include in a panel at which stage. It may be preferable to screen against most members of a particular family upfront, or it may be preferable to pick individual members as representatives and only screen other mem-bers if there is activity in the initial screen.Ion channels. The feasibility of developing or running an assay also has an influence on the original selection of targets. Owing to their prominent physiological roles in cellular excitation, drug interactions with ion channels have long been associated with undesired effects. It can be challenging to develop pharmacologically relevantassays for voltage-gated ion channels that are capable of screening large numbers of compounds. These assays require high-throughput electrophysiological techniques and must capture various modes of action and different channel states. The availability of instruments to perform high-throughput electrophysiology 19 screening has enabled some of these targets to be included in the safety pharmacology profiling repertoire. Some examples include cardiac voltage-gated sodium channel subunit α (Nav1.5), voltage-gated calcium channel subunit α (Cav1.2) as well as potassium voltage-gated channel KQT-like member 1 (KCNQ1) co-expressed with minimal potassium channel (MinK; also known as KCNE1). However, these assays are often run in separate dedi-cated laboratories and at a later stage of the drug development process. Assessments of hERG channel activity are sometimes run as a separate assay owing to the exceptionally high hit rate at this target and the conse-quently higher number of compounds that need profiling. Ligand-gated ion channels such as 5-HT 3, GABA (γ-aminobutyric acid), NMDA (N -methyl-d-aspartate) and nicotinic acetylcholine receptors are often amenable to the development of binding assays and are included in the minimal panel.Enzymes. Following the withdrawal of rofecoxib from the market and the con-sequent concerns about the increased risk of heart attack and stroke associated with long-term use of cyclooxygenase 2 (COX2)-specific inhibitors, all four compa-nies screen against this enzyme, and some also screen against COX1.Monoamine oxidase (MAO) inhibitors were extensively used for treating psychiatric and neurodegenerative disorders, but their use was severely limited by the occurrence of centrally mediated side effects. In par-ticular, MAO inhibitors induce hypertensive crisis when combined with pressor amines — for example, tyramine, which is present in cheese (leading to the term ‘cheese effect’ being coined for this interaction). Moreover, combining MAO inhibitors with other serotonin agonists or selective serotonin reuptake inhibitors also induces a severe risk of a life-threatening serotonin syn-drome. Consequently, MAO isoforms are also included by all four companies in their profiling panels.Most of the phosphodiesterase 4 (PDE4) inhibitors developed so far, in particular for the treatment of asthma and chronic obstructive pulmonary disease, have failed to reach the market owing to on-target adverse effects such as emesis; therefore, PDE4 is also included in the profiling panels. PDE3 is another phosphodiesterase that is included in the profiling panel. PDE3 inhibitors such as milrinone have been used as cardiotonic agents for patients who have suffered from heart failure; these drugs have shown benefi-cial effects on relieving symptoms, but their long-term use (longer than 48 hours) in patients with severe congestive heart failure is associated with pro-arrhythmic activities and increased mortality. The other enzyme included in the panel is acetylcholinesterase (AChE). AChE inhibition causes side effects of varying severity, partly depending on the reversibility of inhibition, and secondary mechanism-of-action studies to determine reversibility may be performed on com-pounds that have high activity against this target.As with cardiac ion channels, kinase screening is an example of how the four different companies similarly developed a procedure whereby kinase screening panels operate semi-independently from the in vitro pharmacology panels. Some com-panies have selected a handful of kinases (between four and ten) to act as ‘sentinel’ representatives of the family. Hits in these sentinel kinases trigger screening in wider kinase panels. There was little overlap in the sentinel kinases chosen by the four companies; indeed, there was only a single common kinase in three out of the four companies: lymphocyte-specific protein tyrosine kinase (LCK).The lack of overlap among the four com-panies probably reflects the relative lack of understanding regarding the implications of modulating specific kinase activity.For instance, there is evidence of cardiotoxicity associated with the clinical use of some poorly selective kinase inhibitors, but infor-mation on the individual kinases responsible is often based on knockout mice or trans-genic models, which could be misleading 20,21. However, for some kinases there is evidence of potential safety liabilities due to known human genetic mutations, and in particular there are now a few highly specific kinase-directed antibodies on the market (some with black box warnings), which provide more direct evidence for specific kinases that are responsible for serious ADRs. We recommend that LCK should not be the only kinase in the panel; rather, a small selec-tion of kinases should be included. However, the challenge is to identify suitable in vitro kinase profiling assays that will be predictive for clinical adverse effects 22. These kinases should then be added into the profile.P E R S P E C T I V E SP E R S P E C T I V E S Table 1 | Recommended targets to provide an early assessment of the potential hazard of a compound or chemical seriesP E R S P E C T I V E STable 1 (cont.) | Recommended targets to provide an early assessment of the potential hazard of a compound or chemical seriesTable 1 (cont.) | Recommended targets to provide an early assessment of the potential hazard of a compound or chemical series1ACO, cardiac output; CVS, cardiovascular system; DDI, drug–drug interaction; ECG, electrocardiogram; GABAA , γ-aminobutyric acid type A; GI, gastrointestinal;HR, heart rate; NA, not applicable; NMDA, N-methyl-d-aspartate; REM, rapid eye movement; SCID, severe-combined immunodeficiency. *Hit rates were determined at 10 μM. ‘Low’ corresponds to <1% hit rate; ‘medium’ corresponds to 1–5% hit rate; ‘high’ corresponds to 5–20% hit rate; ‘very high’ corresponds to >20% hit rate.‡Targets that were included in the panels of all four companies. §The references cited are key references giving details of some of the main adverse drug reactions (ADRs) for each target, but not all of the ADRs listed are mentioned in the cited publications.P E R S P E C T I V E STransporters. Neurotransmitter transporters are important drug targets; however, many are associated with safety liabilities such as increases in blood pressure and abuse liability, and thus three (dopamine, noradrenaline and serotonin transporters) are includedin the minimal in vitro pharmacological pro-filing panel. Transporters that are involved in drug secretion or uptake (for example, liver-specific organic anion transporter 1 (LST1); also known as OATP1B1) or drug-metabolizing enzymes (for example, the cytochrome P450 enzymes) are often tested in separate departments (that is, those that assess the ADME (absorption, distribution, metabolism and excretion) properties of the candidate drugs). These departments have different workflows and so these types of transporters are not included in this phar-macological profiling panel.Nuclear receptors. Nuclear receptors are represented by two targets: the androgen receptor and the glucocorticoid receptor. Neither androgenic nor anti-androgenic effects would be desirable properties in most drugs, whereas interactions withthe glucocorticoid receptor could cause immuno d eficiency, impaired glucose toler-ance, muscle wasting and hypertension. Profiling methods and testing strategies In spite of the similar make-up of the target panels screened by the different pharma-ceutical companies, different technical approaches have been adopted in establish-ing assays for these targets, particularly in the use of ligand binding and functional assays. Typically, binding assays utilize puri-fied preparations of membrane or protein from recombinant or tissue sources express-ing the target and a labelled high-affinity ligand that incorporates either a radioisotope or fluorescent probe. Compounds are tested for their ability to displace the binding of this labelled ligand. Functional assays measure activation, inhibition or modulation of the actual activity of a target, either in a purified preparation or expressed in a host cell. They encompass many different techniques, often with several approaches per target class. GPCR assays generally rely on the meas-urement of second messenger (calcium or cyclic AMP) release or 35S-labelled GTPγS binding, although numerous alternative approaches are possible23. For ion channels, technologies allowing automated electro-physiological measurements of channels expressed in a host cell (for example, using so-called ‘population patch-clamping’) are utilized24,25. Enzymes, including kinases, may be purified and assayed by incubationwith substrates or products that have clearread-outs. Nuclear receptors are usuallyexpressed in cells in combination with genereporters, although biochemical assays usingtime-resolved fluorescence resonance energytransfer (TR-FRET) and fluorescence polari-zation can also be used. BOX 2 outlines someof the factors that influence the use of func-tional and binding assays in the context ofin vitro pharmacological profiling, and sum-marizes the different information generated.Ultimately, these approaches are com-plementary and all four companies rely ona combination of both ligand binding andfunctional assays to establish effective assaypanels26. All assays included in early profilingpanels, binding or functional, need to berobust (that is, they have to produce reliableand reproducible results over years) andthey have to have a predictive value forsafety. It can be challenging to achieve thelevel of robustness required when utilizingcell-based functional assays. Regardless ofwhether binding or functional assays areused as the first-pass test, care needs to betaken to confirm activity and specificity,as with any hit in a high-throughput assay.This is often achieved with a secondaryassay using the complementary technology(that is, a binding assay followed by afunctional assay, or a functional assayfollowed by a binding assay).Ideally, initial testing should be per-formed at multiple test concentrationsto allow a direct estimation of the AC50.This provides a faster and more reliableresult than if testing is performed initially ata single test concentration (typically 10 μMin duplicate) and only the hits are followedup by performing concentration–responsecurves. However, the former strategy canhave a higher cost, especially for targets witha low hit rate, and this has to be calculatedagainst the benefits.With robust binding or enzymatic assays,it is possible to estimate the IC50from singleconcentration data in which the percentageinhibition is between 20% and 80%, andthis can provide the minimum informationrequired for early hazard identification andelimination of off-target activity at a lowercost. However, this approach is not recom-mended when quantitative assessment ofsafety margins is required or if the percent-age inhibition is greater than 85%. If thedata are intended for regulatory submission,it is advisable to explore the concentration–response relationship and generatequantitative data (such as Ki, IC50or EC50)P E R S P E C T I V E S。

新型抗精神障碍药鲁拉西酮个体化用药研究进展

广东药科大学学报Journal of Guangdong Pharmaceutical University Sep,2023,39(5)新型抗精神障碍药鲁拉西酮个体化用药研究进展陈玉清，杨烨，戴丽静，温预关（广州医科大学附属脑科医院Ⅰ期临床研究室，广东广州510370）摘要：鲁拉西酮是一种新型抗精神障碍药（第二代抗精神病药SGA），是多巴胺2型D2和血清素5-HT2A和5-HT7受体完全拮抗剂和部分5-HT1A受体激动剂。

鲁拉西酮对精神分裂症患者的精神病症状、感情症状、认知症状的治疗效果明显，同时对患者体重、代谢影响水平较小，具有良好的安全性及耐受性，广泛应用于临床治疗。

本文综述了鲁拉西酮的作用机制和药动学、群体药动学、安全性、有效性及耐受性、治疗药物检测、基因多态性、药物相互作用研究现状，以期为后续研究及临床个体化用药提供参考。

关键词：鲁拉西酮；药代动力学；治疗药物检测；群体药代动力学；基因多态性；个体化用药中图分类号：R971文献标识码：A文章编号：2096-3653（2023）05-0126-09DOI:10.16809/ki.2096-3653.2023072502Progress on individualized dosing of lurasidone,a new antipsychotic drugCHEN Yuqing,YANG Ye,DAI Lijing,WEN Yuguan*(Phase I Clinical Laboratory of Brain Hospital Affiliated to Guangzhou Medical University,Guangzhou510370,China) *Corresponding author Email:*******************Abstract:Lurasidone is a novel antipsychotic(second-generation antipsychotic SGA),which is a completeantagonist of dopamine type2(D2)and serotonin5-HT2Aand5-HT7receptors and a partial5-HT1Areceptoragonist.It is widely used in the clinical treatment of patients with schizophrenia because of its significant efficacy in psychotic,affective,and cognitive symptoms,and its low effect on body mass and metabolism,as well as its good safety and tolerability.Based on this,the review focuses on the mechanism of action and pharmacokinetic studies,population pharmacokinetics,safety,efficacy and tolerability,therapeutic drug testing,gene polymorphism,and drug interaction studies of lurasidone,with the aim of providing reference for follow-up studies and individualized clinical treatment.Key words:lurasidone;pharmacokinetics;therapeutic drug monitoring;population pharmacokinetics;genetic polymorphisms;individual medication鲁拉西酮（lurasidone）是新型第2代抗精神障碍药之一，由日本住友制药研制。

文献翻译--陈建领

外文资料译文多晶体金属多层膜的变形机制图1引言在塑性方面金属多层膜代表了一种探索长度尺度的理想工具。

他们还提供了用控制界面和结构来生产接近理论强度的合成材料的机会。

一些作者依据Hall-Petch和Orowan强化机制探索了长度尺度的影响。

Embury和Fisher 早期绘制的珠光体机制图表明，Hall-Petch在单相金属晶粒细化加强模型也适用于有作为阻碍距离的界面间隔的双相材料。

更多最近的研究，例如Embury-Hirth，Anderson等，Chu和Barnett和Nix 表明在纳米级多层膜的力学行为可能受单个位错行为（Orowan的层间位错弯曲模型），而不是逆着界面的堆积位错。

Masumura等人的另一项最近的研究表明在单相纳米材料中晶粒尺寸低于临界晶粒尺寸，基础的扩散机制，如Coble蠕变可能会执行，并可能导致伴随晶粒细化的软化。

为简便起见，通常这些模型，无论是有单晶成份层的多层膜或多层膜的单相细晶材料都很发达。

了解多晶多层膜的力学性能构成由于两个层厚度和面晶粒的尺寸可能影响屈服强度的一个额外的复杂性。

虽然平面晶粒的大小可能改变层厚度，没有普遍的关系使我们在只知道层厚度时能够计算出晶粒尺寸，反之亦然。

这些参数之间的关系通常是通过详细的微观结构的特性来决定。

因此，对于给定的多晶金属多层膜，我们如何获取关于层厚度和晶粒大小值执行不同的组合，变形机制见解？在本次调查中，我们提出一个简单的分析，使我们在这些不同的机制运作时能够获得微观尺度的极限值。

我们提出的结论在层厚度和晶粒大小的二维图形的形式，包括不同的变形机制的运作。

这些图形的目的是用于解释尺寸强化或多层膜软化机制，相同的方法，像Ashby的变形温度和压力机制图一样，都是依赖金属变形行为。

Frost试图扩展Ashby的变形机制图，他的的铝薄膜变形机制图表明，由于薄膜层的更高的循环应力，预测应变率数量级比实测应变率高几个数量级。

因此，需要开展更多工作纳入薄膜和大块多晶体的变形行为的基本差别，机制图是压力，温度和微观尺度的一个函数。

麻醉英语考核 -回复

麻醉英语考核-回复An Introduction to Anesthesia-Induced Sedation and Its Importance[Introduction]Anesthesia-induced sedation is a crucial aspect of modern medical practices, ensuring patients are comfortable and pain-free during surgical procedures. This article aims to provide a comprehensive understanding of anesthesia-induced sedation, including its definition, types, benefits, potential risks, and the role of anesthesiologists in this process.[Definition and Types]Anesthesia-induced sedation refers to the administration of medications to induce relaxation, calmness, and reduce anxiety or discomfort in patients undergoing medical procedures. It plays a vital role in not only maintaining the patient's comfort but also providing optimal conditions for the surgeon to perform the required intervention.There are several types of anesthesia-induced sedation, categorized based on their intensity and effects on patients. These include minimal sedation, moderate sedation, deep sedation, and general anesthesia. Minimal sedation involves the administration of medications that promote relaxation but allow the patient to remain awake and respond to stimuli. Moderate sedation induces a state of depression of consciousness, where patients may require minimal stimulation to maintain breathing and respond to commands. Deep sedation results in a state where patients are not easily aroused but can still respond to painful stimuli. General anesthesia brings patients into a state of total unconsciousness, requiring assistance with breathing and complete absence of pain perception.[Benefits of Anesthesia-Induced Sedation]The primary benefit of anesthesia-induced sedation is the comfort it provides to patients. By reducing anxiety and pain, sedation promotes a relaxed state, enhancing the overall patient experience. Additionally, anesthesia-induced sedation enables medical professionals to perform procedures more effectively and efficiently by minimizing patient movements, facilitating access tothe surgical site, and ensuring optimal conditions for the surgeon.Furthermore, sedation can aid in the management of patients with underlying medical conditions that require interventions. For example, individuals with respiratory conditions may experience induced suppression of their drive to breathe, which can be counteracted with appropriate sedation techniques to alleviate discomfort. Therefore, anesthesia-induced sedation is a crucial component of ensuring the safety and well-being of patients during surgical procedures.[Potential Risks and Precautions]Although anesthesia-induced sedation is considered safe and routine in modern medicine, it carries potential risks that necessitate precautions to minimize complications. Some risks include adverse reactions to sedative medications, fluctuations in blood pressure or heart rate, respiratory depression, allergies, and even rare instances of anaphylactic reactions.To minimize these risks, anesthesiologists are trained professionals who possess an in-depth understanding of physiology andpharmacology. They conduct thorough pre-operative evaluations, considering the patient's medical history, current condition, and any potential contraindications. This information allows them to adjust medication dosages and monitor patients closely during the procedure to ensure their safety.Anesthesiologists also employ advanced monitoring techniques, such as electrocardiography (ECG), pulse oximetry, noninvasive blood pressure monitoring, and capnography. These monitoring tools provide real-time data on the patient's vital signs, enabling prompt recognition and intervention in case of any complications.[The Role of Anesthesiologists]Anesthesiologists play a vital role in the administration of anesthesia-induced sedation. Their responsibilities extend beyond merely providing sedative medications. They are involved in the entire perioperative care of patients, which includes preoperative evaluations, creating individualized anesthesia plans, selecting appropriate medications, closely monitoring patients during the procedure, and managing post-operative recovery.Furthermore, anesthesiologists possess the necessary knowledge and skills to handle any potential complications that may arise during anesthesia-induced sedation. They are trained in advanced airway management techniques, resuscitation, and critical care, allowing them to respond efficiently in emergency situations.[Conclusion]Anesthesia-induced sedation is a fundamental aspect of modern medical practices that ensure patients' comfort and welfare during surgical procedures. With the advancements in pharmacology, monitoring techniques, and the expertise of anesthesiologists, this process has become increasingly safe and effective. By providing relaxation, reducing anxiety, and maintaining patient stability, anesthesia-induced sedation significantly contributes to improving surgical outcomes and patient experiences.。

人教版高中英语选择性必修第二册课后习题 Unit 1 第一单元测评卷

第一单元测评第一部分听力(共两节,满分30分)第一节(共5小题;每小题1.5分,满分7.5分)听下面5段对话。

每段对话后有一个小题,从题中所给的A、B、C三个选项中选出最佳选项。

听完每段对话后,你都有10秒钟的时间来回答有关小题和阅读下一小题。

每段对话仅读一遍。

1.What will the woman do this coming weekend?A.Call on her classmates.B.Direct traffic.C.Go hiking.2.Where is the woman going?A.To her new flat.B.To the shop.C.To her company.3.When did the woman buy her own car?st week.st month.st year.4.What does the woman do now?A.A school teacher.B.A tourist guide.C.A university student.5.How does John look?A.Excited.B.Satisfied.C.Worried.第二节(共15小题;每小题1.5分,满分22.5分)听下面5段对话或独白。

每段对话或独白后有几个小题,从题中所给的A、B、C三个选项中选出最佳选项。

听每段对话或独白前,你将有时间阅读各个小题,每小题5秒钟;听完后,各小题将给出5秒钟的作答时间。

每段对话或独白读两遍。

听第6段材料,回答第6、7题。

6.What will the weather be like the day after tomorrow?A.It is rainy.B.It is sunny.C.It is windy.7.What has the man been doing these days?A.Reading books.B.Playing football.C.Going cycling. 听第7段材料,回答第8至10题。

英语论文文献综述(描述研究现状)

General descriptions of the relevant literature：A considerable amount of literature has been published on X。

These studies 。

There is a large volume of published studies describing the role of ...。

The first serious discussions and analyses of X emerged during the 1970s with .。

The generalisability of much published research on this issue is problematic.What we know about X is largely based upon empirical studies that investigatehow .。

.During the past 30 years much more information has become available on 。

...In recent years, there has been an increasing amount of literature on .。

..A large and growing body of literature has investigated .。

.General reference to previous research/scholarly activity （usually more than one author)Many historians have argued that .。

. (eg。

Jones, 1987; Johnson，1990；Smith，1994）Numerous studies have attempted to explain 。

药学英语Unit 6 Text A 注释及译文

Drug Discovery and Natural Products It may be argued that drug discovery is a recent concept that evolved from modern science during the 20th century, but this concept in reality dates back many centuries, and has its origins in nature. On many occasions, humans have turned to Mother Nature for cures, and discovered unique drug molecules. Thus, the term natural product has become almost synonymous with the concept of drug discovery. In modem drug discovery and development processes, natural products play an important role at the early stage of "lead" discovery, i.e. discovery of the active (determined by various bioassays) natural molecule, which itself or its structural analogues could be an ideal drug candidate.1.origin ['ɔridʒin] n.起点，端点; 来源；出身, 血统.2.Synonymous [sɪ'nɔnəməs]adj.同义的，类义的.3.i.e. [,aɪ'i:] <拉> abbr. (=id est) 即，换言之.4.candidate ['kændidit] n.申请求职者, 候选人；报考者；候选物.有人可能认为药物发现是一个20世纪才出现的、来源于现代科学的新概念，但是事实上这个概念是源于自然界的，可以追溯到许多个世纪以前。

西洋参提取物对癌症疲劳的研究【翻译、题库

安全有效的预防和治疗癌症的干预疲劳（CRF）是因为它仍然是最有—本科以上学历，令人不安的，和活动限制症状的幸存者经验，在短期和长期（1–6）。

患病率正在接受化疗的患者的疲劳是之间的59%和96%，在接受放射治疗的患者，65%和100%之间（4）。

疲劳可以持续5到10 年诊断和治疗后（2,3）。

CRF深刻和负影响患者的生活质量和干扰日常功能（5,6）。

此外，大量疲劳帐户在整体生活质量的差异金额，超过40%的差异归因于疲劳（1）。

有一个缺乏证据支持的疗效的药—macologic干预。

兴奋剂是最研究慢性肾功能衰竭的药物干预。

解放军八—安慰剂随机对照试验评价哌醋甲酯或相关试剂已经完成，但被试（8）负（9–15）。

尽管受欢迎，新的兴奋剂，如modafanil，也没有被发现是有效的随机对照试验（16）。

此外，其他中枢神经—我们的系统（例如，多奈哌齐，帕罗西汀）有负面的结果的日期（17,18）。

膳食补充剂是一种流行的自我救济有没有有效的治疗症状的患者—彪；CRF也不例外。

辅酶Q 10，左旋肉碱，瓜拉那，人参已用于疲劳和随后的研究。

基于其在细胞能量生产的作用，辅酶Q 10和左旋肉碱在安慰剂对照试验评价慢性肾功能衰竭；他们发现没有比安慰剂更有效（19,20）。

瓜拉纳具有支持数据从第二阶段的安慰剂对照—控制试验（21），值得进一步研究。

虽然许多植物已经被通过民俗通过使用传统的补救疲劳，不可能尽可能多的全球声誉和利益为人参享受。

在中医中，人参通常被视为一种“适应原，“一种物质能够帮助恢复平衡的身体带回一点平衡（22）。

有两个主要种类的人参，亚洲（人参）和美国（西洋参）（22、23）。

两有一个共同的活性成分的混合物，最重要的被ginsensosides。

种人参之间，有不同量，强度，和品种的ginsensosides（22,24,25）。

客观的证据支持，人参可有利于疲劳来自临床前数据。

具体而言，在体外数据显示抗炎和皮质醇的调节影响（26–28）与当前建立的生理一致—OGY CRF。

开启片剂完整性的窗户(中英文对照)

开启片剂完整性的窗户日本东芝公司，剑桥大学摘要：由日本东芝公司和剑桥大学合作成立的公司向《医药技术》解释了FDA支持的技术如何在不损坏片剂的情况下测定其完整性。

太赫脉冲成像的一个应用是检查肠溶制剂的完整性，以确保它们在到达肠溶之前不会溶解。

关键词：片剂完整性，太赫脉冲成像。

能够检测片剂的结构完整性和化学成分而无需将它们打碎的一种技术，已经通过了概念验证阶段，正在进行法规申请。

由英国私募Teraview公司研发并且以太赫光（介于无线电波和光波之间）为基础。

该成像技术为配方研发和质量控制中的湿溶出试验提供了一个更好的选择。

该技术还可以缩短新产品的研发时间，并且根据厂商的情况，随时间推移甚至可能发展成为一个用于制药生产线的实时片剂检测系统。

TPI技术通过发射太赫射线绘制出片剂和涂层厚度的三维差异图谱，在有结构或化学变化时太赫射线被反射回。

反射脉冲的时间延迟累加成该片剂的三维图像。

该系统使用太赫发射极，采用一个机器臂捡起片剂并且使其通过太赫光束，用一个扫描仪收集反射光并且建成三维图像(见图)。

技术研发太赫技术发源于二十世纪九十年代中期13本东芝公司位于英国的东芝欧洲研究中心，该中心与剑桥大学的物理学系有着密切的联系。

日本东芝公司当时正在研究新一代的半导体，研究的副产品是发现了这些半导体实际上是太赫光非常好的发射源和检测器。

二十世纪九十年代后期，日本东芝公司授权研究小组寻求该技术可能的应用，包括成像和化学传感光谱学，并与葛兰素史克和辉瑞以及其它公司建立了关系，以探讨其在制药业的应用。

虽然早期的结果表明该技术有前景，但日本东芝公司却不愿深入研究下去，原因是此应用与日本东芝公司在消费电子行业的任何业务兴趣都没有交叉。

这一决定的结果是研究中心的首席执行官DonArnone和剑桥桥大学物理学系的教授Michael Pepper先生于2001年成立了Teraview公司一作为研究中心的子公司。

TPI imaga 2000是第一个商品化太赫成像系统，该系统经优化用于成品片剂及其核心完整性和性能的无破坏检测。

An Investigation of the Effects of the

An Investigation of the Effects of the Core Protein Telomerase Reverse Transcriptase on Wnt Signaling in Breast Cancer CellsImke Listerman,Francesca S.Gazzaniga,Elizabeth H.BlackburnDepartment of Biochemistry and Biophysics,University of California at San Francisco,San Francisco,California,USATelomerase canonically maintains telomeres,but recent reports have suggested that the core protein mammalian telomerase reverse transcriptase(TERT)component,together with the chromatin remodeling factor BRG1and␤-catenin,may also bind to and promote expression of Wnt target genes.However,this proposed noncanonical role of TERT in Wnt signaling has been con-troversial.Here,we investigated the effects of human TERT(hTERT)on Wnt signaling in human breast cancer lines and HeLa cells.We failed toﬁnd evidence for physical association of hTERT with BRG1or␤-catenin;instead,we present evidence that anti-FLAG antibody cross-reactivity properties may explain the previously reported interaction of hTERT with␤-catenin.Fur-thermore,altering hTERT levels in four different breast cancer cell lines caused minimal and discordant effects on Wnt target and Wnt pathway gene expression.Although hTERT’s role in Wnt signaling was addressed only indirectly,no signiﬁcant repre-sentation of Wnt target genes was detected in chromatin immunoprecipitation-sequencing(ChIP-seq)and chromatin isolation by RNA puriﬁcation and sequencing(ChIRP-seq)loci cooccupied in HeLa S3cells by both BRG1and hTR.In summary,our evi-dence fails to support the idea of a biologically consistent hTERT interaction with the Wnt pathway in human breast cancer cells, and any detectable inﬂuence of hTERT depended on cell type and experimental system.T he mammalian telomerase ribonucleoprotein complex adds TTAGGG repeats to telomeres,the ends of linear chromo-somes.The core human telomerase contains the catalytic reverse transcriptase protein component(hTERT)and the telomerase RNA(called hTR,hTER,or hTERC)that provides the template for telomeric DNA synthesis(1).In most human somatic cells, telomerase expression is very low.In contrast,telomerase expres-sion is upregulated in many human cancer cells and stem cells(2). In human cancer cells,the degree of telomerase expression seems higher than would appear necessary solely for maintaining telo-mere length.In fact,many studies suggest telomere-independent roles for telomerase.We and others have shown that overexpres-sion of TERT protects cells in culture from apoptosis indepen-dently of the telomere-lengthening properties of telomerase(3–5).Furthermore,overexpression of mouse and human TERT promotes cell proliferation in stem,normal,and cancer cell lines (6–11).Experiments employing overexpression or reduced ex-pression of hTERT in cells in culture have suggested roles for hTERT in controlling expression of growth factor response and other genes(9,12).Gene expression changes have been reported to occur as soon as1week after ectopic hTERT overexpression(9). Taken together,these results strongly suggest nontelomeric roles for telomerase;however,the mechanisms by which telomerase might protect against apoptosis and promote proliferation remain largely unknown.Some previous studies have linked TERT expression and Wnt/␤-catenin signaling,here referred to as Wnt signaling(13–15). The Wnt signaling pathway plays a central role in development, stem cell renewal,and cancer.In the absence of Wnt signaling, cytoplasmic␤-catenin is bound by destruction complex proteins, including AXIN,adenomatous polyposis coli(APC),and glyco-gen synthase kinase3beta(GSK3B).Consequently,␤-catenin is phosphorylated and degraded by the ubiquitin-proteasome path-way.When secreted Wnt proteins bind to Frizzled and low-den-sity lipoprotein receptor-related proteins(LRPs)at the plasma membrane,a signal is transduced to destabilize the␤-catenin de-struction complex.␤-Catenin can then translocate to the nucleus, where it complexes with T-cell factor/lymphoid enhancer factor (TCF/LEF)transcription factors to promote target gene transcrip-tion(16).The Wnt pathway has been previously shown to upregu-late telomerase in mouse mammary tumors and human cells(17, 18).Furthermore,␤-catenin may contribute to telomerase up-regulation in stem and cancer cells by directly regulating TERT expression via binding to the TERT promoter in complex with Klf4,as previously reported in a study of mouse adult stem cells and human carcinoma lines NTera2and SW480(15).Reciprocally,Park et al.previously suggested that TERT ex-pression promotes Wnt signaling(13).In that study,TERTϪ/Ϫknockout mice in theﬁrst generation were reported to have devel-opmental defects such as homeotic transformations of the vertebrae.Such defects,occurring before the onset of signiﬁcant telomere shortening,resembled effects of aberrant Wnt signaling. Those authors additionally reported protein-protein interactions between hTERT and the chromatin remodeling factor BRG1and between hTERT and␤-catenin.It was also reported that TERT overexpression upregulated expression of a Wnt luciferase re-porter in TERTϪ/Ϫand TRϪ/Ϫmouse embryonicﬁbroblasts (MEFs)and humanﬁbroblast(BJ)cells and that,in SW-13and HeLa cancer cells,TERT overexpression hyperactivated a Wnt signaling reporter in a BRG1-dependent manner(13).Consistent with these results,Hrdlickováet al.reported increased prolifera-tion and a slight but signiﬁcant increase in Wnt reporter activation Received28June2013Returned for modiﬁcation6August2013Accepted4November2013Published ahead of print11November2013Address correspondence to Elizabeth H.Blackburn,Elizabeth.Blackburn@.I.L.and F.S.G.contributed equally to this article.Copyright©2014,American Society for Microbiology.All Rights Reserved.doi:10.1128/MCB.00844-13 Molecular and Cellular Biology p.280–289January2014Volume34Number2upon overexpression of either hTERT or a catalytically incompe-tent hTERT splice variant,in both U2OS(telomerase-deﬁcient) and HeLa(telomerase-positive)cell lines(19).BRG1has been reported to bind to␤-catenin and to promote␤-catenin target gene expression(20,21).Because many growth-promoting genes are␤-catenin targets and because Wnt signaling plays an impor-tant role in self-renewal,proliferation,and survival,these reports suggested that TERT,in concert with BRG1,might promote cell proliferation via Wnt signaling.An inﬂuence of TERT on Wnt signaling has not been consis-tently reproduced in other experimental settings.Strong et al.did not detect homeotic transformations or diminished Wnt re-porter activity in TERTϪ/Ϫknockout mice or mouse embry-onicﬁbroblasts(MEFs)derived from these mice(22).The dis-crepancies between the two mouse TERTϪ/Ϫknockout studies could have been due to slightly different experimental condi-tions,such as different mouse backgrounds and/or Wnt signal-ing activators(13,22).Alternatively,the TERT overexpression system that identiﬁed a Wnt pathway interaction(13)may not produce a biologically relevant phenotype.While Strong et al. disputed a TERT/Wnt signaling interaction in mice and MEFs (22),they did not address a possible TERT/Wnt signaling in-teraction in human cancer cells.The primary aim of the present study was to determine whether hTERT promotes or otherwise affects Wnt signaling in cultured human breast cancer cells.Wnt signaling is often dys-regulated in breast cancer(23).Furthermore,either TERT over-expression or Wnt activation leads to mammary tumorigenesis in mice(23,24).We therefore focused on breast cancer cell lines to further study the potential for biologically relevant TERT/Wnt signaling interactions.MATERIALS AND METHODSCell lines.HeLa cells were purchased from the American Type Culture Collection(ATCC)and grown in Dulbecco’s modiﬁed Eagle’s medium (DMEM)with10%fetal bovine serum(FBS),1%penicillin-streptomy-cin,and1%GlutaMax(Invitrogen).HCC3153,HCC1806,SUM149PT, and MCF10A cells were obtained from the laboratory of Joe W.Gray, Oregon Health and Sciences University.HCC3153and HCC1806cells were grown in RPMI medium with10%FBS,1%penicillin-streptomycin, and1%GlutaMax(Invitrogen).SUM149PT cells were grown in Ham’s F-12medium with5%FBS,0.01mg/ml insulin,500ng/ml hydrocorti-sone,and1%penicillin-streptomycin.MCF10A cells were grown in DMEM–F-12with5%horse serum,20ng/ml epidermal growth factor, 100ng/ml cholera toxin,0.01mg/ml insulin,500ng/ml hydrocortisone, and1%penicillin-streptomycin.All cell lines were grown at37°C with5% CO2.Light microscopy.The HCC3153,HCC1806,SUM149PT,and MCF10A cell lines were grown on chamber slides(Lab-Tek II154526)and treated with25mM LiCl or200ng/ml Wnt3a(5036-WN-010/CF;R&D Systems)for4h.Cells wereﬁxed in2%paraformaldehyde–phosphate-buffered saline(PBS)and permeabilized with0.5%NP-40–PBS.Immu-nostaining was performed with anti-␤-catenin antibody clone14(BD Biosciences)followed by secondary Alexa Fluor488(Molecular Probes). DNA was visualized with4=,6-diamidino-2-phenylindole(DAPI;Invitro-gen).Images were acquired in0.5-␮m increments using a Deltavision RT deconvolution microscope(Applied Precision)with a100ϫ/1.40N PlanApo objective(Olympus).Images were deconvolved,Z-projected in Softworx(Applied Precision),and then adjusted for brightness and con-trast in FIJI(25).cDNA generation and qPCR.Total RNA was extracted with a Qiagen RNeasy minikit from cells treated with25mM LiCl,200ng/ml Wnt3a,or PBS for4h.cDNA synthesis was performed using2␮g RNA,random hexamers,and SuperScript III(Invitrogen).cDNA was ampliﬁed in10-␮l reaction mixtures containing LightCycler480DNA SYBR green I Master (Roche Applied Science)and a0.5to1␮mol/literﬁnal concentration of each primer using a Light Cycler480instrument(Roche Applied Science). The cycling conditions were95°C for5min,50cycles of95°C for10s and 60°C for20s,and72°C for20s.A melting curve(65to98°C)was gener-ated at the end of each run.Relative expression levels were determined by the2Ϫ⌬⌬CT method(26)and were normalized to GAPDH(glyceralde-hyde-3-phosphate dehydrogenase).AXIN2forward(5=-CATGTTCGTC ATGGGTGTGAACCA-3=)and AXIN2reverse(5=-TGGCTGGTGCAAA GACATAG-3=)and GAPDH forward(5=-CATGTTCGTCATGGGTGTG AACCA-3=)and GAPDH reverse(5=-ATGGCATGGACTGTGGTCATG AGT-3=)primers were used.For Wnt target gene expression analysis,cell lines were transduced with control or wild-type hTERT lentivirus pHR=cytomegalovirus(CMV)-hTERT-internal ribosome entry site (IRES)-PURO and selected with1␮g(HCC1806and SUM149PT)or2␮g (HCC3153)puromycin for3days and allowed to recover for1to2days. Cells were treated with25mM LiCl for6h,following total RNA extraction using an RNeasy kit(Qiagen)and cDNA generation using an RT2First Strand kit(Qiagen)according to the manufacturer’s instructions.A total of84Wnt target genes were measured using quantitative PCR(qPCR) human Wnt signaling target arrays(PAHS-243G;SABiosciences)accord-ing to the manufacturer’s instructions.A minimum cutoff of a2.5-fold change compared to control results was used to determine signiﬁcant gene changes.Wnt luciferase reporter assays.The M50Super TOPFLash and M51 Super FOPFlash luciferase reporter vectors were obtained from Addgene (27).pRL-TK Renilla luciferase was used as an internal control(Invitro-gen).The lentivirus plasmids pBARL(␤-catenin activated reporter lucif-erase)and pfuBARL(mutated pBARL)and pSL9/rLuc(Renilla luciferase) were obtained from the laboratory of Randall Moon,University of Wash-ington(28).Cells were seeded on96-well microplates(655083;Greiner Bio-One),and each well was transiently transfected with0.5pg pRL-TK control along with either50pg SUPER TOPFLash or50pg mutated FOPFLash and with either10pg empty vector or10pg pcDNA3-FLAG-hTERT using X-tremeGene HP(Roche)and treated or not treated with25 mM LiCL for24h,followed by cell lysis with passive lysis buffer(Pro-mega)for10min and analysis using a dual-luciferase reporter assay sys-tem(Promega).Fireﬂy and Renilla luciferases were read with a Veritas Microplate Luminometer(Turner Biosystems).Lentivirus production and transduction were carried out as previously described(28).Stable cell lines expressing reporter Renilla luciferase,pBARL,or pfuBARL lentivirus were generated as reported previously(28),using the same titer of lenti-virus in all cell lines.Then,cell lines were transduced with either control or hTERT lentivirus(28),selected with puromycin,treated with25mM LiCl for24h,and analyzed as described above.Background luciferase readings were subtracted,andﬁreﬂy luciferase values were normalized to Renilla luciferase.RNA interference(RNAi).Lentivirus expressing short hairpin RNAs (shRNAs)against␤-catenin(5=-CCGGAGGTGCTATCTGTCTGCTCT ACTCGAGTAGAGCAGACAGATAGCACCTTTTTT-3=;29),hTERT (5=-GGAGACCACGTTTCAAAAGTCTCTTGAACTTTTGAAACGTGG TCTCC-3=),and scramble shRNA(5=-GTTCTACAACGTAACGAGGTT TCTCTTGAAAACCTCGTTACGTTGTAGAAC-3=;30)was generated as described previously(30).Cells were transduced with shRNA and control vector lentivirus and were selected with1␮g/ml puromycin for3days and then expanded.IP and Western blotting.HeLa cells were transfected with pcDNA3 constructs containing wild-type hTERT with one N-terminal FLAG tag using Lipofectamine2000(Invitrogen)for18h,followed by treatment with25mM LiCl for6h.The following antibodies were used:anti-FLAG (M2;Sigma F3165and F1804),anti-BRG1(H-88;Santa Cruz),anti-␤-catenin(clone14)(610153;Transduction Laboratories),and anti-GAPDH(MAB374;EMD Millipore).Cell lysis and immunoprecipitationhTERT Effects on Wnt Signaling in Breast Cancer281(IP)procedures and immunoblotting were done as described previously (13),using Western Lightning Plus ECL(PerkinElmer)for detection of horseradish peroxidase-conjugated secondary antibodies and Gamma-Bind G Sepharose(GE Healthcare Life Sciences)for preclearing and IP.Bioinformatics analysis.We determined the union of published BRG1-and hTR-enriched regions identiﬁed by chromatin immunopre-cipitation-sequencing(ChIP-seq)and chromatin isolation by RNA puri-ﬁcation and sequencing(ChIRP-seq),respectively,in HeLa S3cells(31, 32)and merged any united regions that were separated byՅ100bp using the ChIPPeakAnno R package(33).Enriched gene sets were obtained through use of the Genomic Regions Enrichment of Annotations Tool (GREAT)(34)on all145genomic regions.Gene ontology(GO)terms were identiﬁed using DAVID(35,36).RESULTSEndogenous Wnt signaling competency varies among the basal breast cancer cell lines SUM149PT,HCC1806,HCC3153,and MCF10A.One immortalized breast cell line with low telomerase activity and three basal breast cancer cell lines with midrange to high levels of telomerase activity(MCF10A[lower telomerase ac-tivity level],HCC3153[midrange telomerase activity level],and SUM149PT and HCC1806[higher telomerase activity levels]) were selected(4).First,to determine the extent of Wnt signaling in the breast cancer cell lines,we induced Wnt signaling with Wnt3a or LiCl.Wnt3a activates the pathway at the cell surface receptor level and speciﬁcally induces␤-catenin signaling by binding to Frizzled and LRP receptors(37).LiCl pharmacologically inhibits GSK3B kinase activity in the cytoplasm,thus leaving␤-catenin un-phosphorylated and stabilized(38).Under control conditions,both SUM149PT and HCC3153cells showed diffuse cytoplasmic and nuclear␤-catenin staining,suggesting that they may have had dysregulated Wnt signaling,which is found frequently in breast cancers(39).Upon Wnt3a orLiCl treatment,SUM149PT and HCC3153cells showed strongernuclear localization of␤-catenin(Fig.1A).In accordance with theincreased nuclear localization of␤-catenin in those cells,bothtreatments increased the expression of the endogenous AXIN2␤-catenin target gene(Fig.1B).In contrast,in HCC1806cells,␤-catenin was largely membrane bound and remained so even after LiCl or Wnt3a treatment(Fig.1A).Consistent with theseﬁndings,HCC1806cells did not signiﬁcantly upregulate AXIN2after Wnt signaling induction(Fig.1B).In MCF10A cells,␤-catenin was also largely membrane bound but showed weak nuclear localization after LiCl or Wnt3A treatment(Fig.1A),and AXIN2was moderately upregulated(Fig.1B).These results sug-gest that HCC1806cells are not competent for Wnt signaling in-duction by LiCl or Wnt3A.We conclude that Wnt signaling can be activated in SUM149PT and HCC3153lines,and somewhat less in MCF10A cells,but at most minimally in HCC1806cells.hTERT overexpression has minimal and nonconcordant ef-fects on Wnt signaling reporters in breast cancer cell lines.Hav-ing established that HCC3153,SUM149PT,and MCF10A but notHCC1806cancer cells can strongly to moderately activate Wntsignaling,we tested whether hTERT overexpression modulatedWnt signaling reporter genes in these lines,as has been reportedfor MEFs,HeLa cells,and U2OS cells(13,19).For independentFIG1Endogenous Wnt/␤-catenin target gene induction varies in breast cancer cell lines.(A)Cell lines SUM149PT(high telomerase),HCC3153(medium telomerase),HCC1806(high telomerase),and MCF10A(low telomerase)were treated with PBS,25mM LiCl,or200ng/ml Wnt3a for4h prior to staining for ␤-catenin(green)and DAPI(blue).(B)The increase in Wnt/␤-catenin target gene AXIN2mRNA expression over that of PBS control-treated cells was measuredby qRT-PCR following activation with Wnt3a or LiCl for4h.Listerman et al. Molecular and Cellular Biologyveriﬁcation,we employed two different Wnt signaling reporter construct systems with multimerized TCF/LEF binding sites driv-ing luciferase expression:the M50Super TOPFlash reporter and its corresponding control M51Super FOPFlash Wnt reporter(27) and the BARL(␤-catenin activated reporter luciferase)/fuBARL (control)system(28).While TOPFlash contains7TCF/LEF bind-ing sites and was transiently expressed via plasmid transfection, BARL contains12TCF/LEF binding sites and was stably inte-grated.Vector or hTERT plasmids were cotransfected together with TOPFlash/FOPFlash in SUM149PT,HCC5313,HCC1806, MCF10A,and HeLa cells,followed by LiCl treatment(Fig.2A).As expected,LiCl treatment strongly increased luciferase activity in the vector-transfected TOPFlash SUM149PT,HCC3153, MCF10A,and HeLa cells.LiCl treatment induced the luciferase activity in HCC1806cells only weakly,with maximum luciferase expression being5-to100-fold lower than in the other four cell lines,consistent with our observations(Fig.1),indicating that HCC1806cells are severely impaired in Wnt signaling.Further-more,only HCC3153cells exhibited a statistically signiﬁcant but mild(ϳ1.4-fold)increase in relative luciferase activity over that of the vector control,while the relative luciferase activity did not change signiﬁcantly compared to that of vector controls in SUM149PT,HCC1806,MCF10A,or HeLa cells.In the pBARL cells,overexpressing hTERT increased luciferase activity over that of the vector control cells only in HCC3153cells(byϳ2-fold) (Fig.2B).Overexpression of hTERT did not detectably change the luciferase activity in SUM149PT,HCC1806,or MCF10A cell lines expressing pBARL.In HeLa cells that stably expressed the TCF/ LEF mutant(control)binding site construct,fuBARL,with hTERT overexpression,we observed anϳ3-fold increase in lucif-erase activity over that of control cells.However,in HeLa cells stably expressing BARL,there was an onlyϳ2-fold increase in luciferase activity in hTERT-overexpressing cells over that of control cells(Fig.2B).Hence,hTERT overexpression activated luciferase expression regardless of the presence or absence of a functional TCF/LEF promoter in HeLa cells.Because hTERT overexpression led to mild hyperactivation of both Wnt signaling reporters only in HCC3153cells and not in the four other cell lines,we conclude that Wnt reporter hyperactivation through hTERT is dependent on the context.Hence,hTERT does not hy-peractivate Wnt reporters universally but instead does so in a cellline-and context-dependent manner.Lack of evidence for hTERT interaction with␤-catenin orBRG1in HeLa cells.Since Park et al.(13)reported that FLAG-hTERT in HeLa cells coimmunoprecipitated(co-IP)with BRG1,aprotein previously reported to interact with␤-catenin(13,20),wealso investigated hTERT/Wnt pathway interactions using HeLacells.To independently verify the previously published results(13),we transiently overexpressed FLAG-hTERT in LiCl-treatedHeLa cells and tested whether BRG1or␤-catenin interacted withFLAG-hTERT by coimmunoprecipitation(co-IP).Interestingly,using the same buffers as described by Park et al.(13)and thenon-afﬁnity-isolated version of anti-FLAG antibody M2(F3165;Sigma)for IP,we observed a strong band migrating slightly slowerthan the␤-catenin band in Western blots(M2lanes in Fig.3A)when the FLAG-IP Western blot was stained with the␤-cateninantibody.Importantly,we also observed the same band in similarquantities independently of whether FLAG-hTERT was expressedin the HeLa cells(M2lanes in Fig.3A)and using a variety ofwashing procedures in the IP and Western blot experiments.Weextended these experiments using the afﬁnity-isolated anti-FLAGM2antibody(F1804;Sigma),which,while it did not enrich forthis background band,instead detected another band of the ex-pected size for␤-catenin at low levels(1.7-fold over IgG control IPresults)that again were identical regardless of whether FLAG-hTERT was expressed(Fig.3B).We veriﬁed the identity of thiscross-reacting coimmunoprecipitated protein band as␤-cateninby RNAi:reducing␤-catenin expression produced correspondingreductions in the intensity of the band pulled down by the afﬁnity-puriﬁed M2antibody co-IP experiments(Fig.3C).Thus,we didnot detect a signiﬁcant or speciﬁc interaction between hTERT and ␤-catenin in HeLa cells above the background signals caused by anti-FLAG antibody ing the same antibodiesand IP buffers as Park et al.(13),we were also unable to detect aninteraction between overexpressed FLAG-hTERT and endoge-nous BRG1,despite obtaining high signals corresponding toFLAG-hTERT itself with the FLAG antibodies used(Fig.3B).In addition,we were able to detect only a weak interaction,atbest,between endogenous levels of BRG1and␤-catenin;suchan interaction has previously been reported only in a BRG1FIG2Effect of hTERT overexpression on two Wnt/␤-catenin reporters.(A)SUM149PT,HCC3153,HCC1806,MCF10A,and HeLa cells were transiently transfected with pRL-TK Renilla luciferase vector(internal control),hTERT or vector,and M50SuperTOPFlash or M51SuperFOPFlash reporter vectors and treated or not treated with25mM LiCl for24h prior to luciferase measurement.*,PϽ0.05.(B)Cell lines were transduced with pSL9/rLuc,pBARL,or pfuBARL and either hTERT or vector control lentivirus and selected for stable expression prior to LiCl treatment and luciferase measurement.hTERT Effects on Wnt Signaling in Breast Cancer 283overexpression context(20).We conclude,ﬁrst,that the en-dogenous expression levels of BRG1in HeLa cells were too low to detect strong interactions with␤-catenin in our experi-ments,second,that M2anti-FLAG antibody cross-reacts with a protein with a gel mobility close to that of␤-catenin,and third, that the interaction of FLAG-hTERT with BRG1or with ␤-catenin was not signiﬁcantly above background IP levels.BRG1and hTR do not colocalize at Wnt target genes.In hu-man cells,the hTERT protein and the telomerase RNA hTR,to-gether with additional proteins,assemble to form the telomerase ribonucleoprotein complex,although it has not been determined what fractions of the total levels of hTERT and hTR exist in these complexes.Previously,Chu et al.(31)used whole-genome chro-matin isolation by RNA puriﬁcation(ChIRP)in HeLa S3cells to detect hTR associated with chromatin at2,198genomic locations in HeLa S3cells.Those authors additionally reported that the hTR-bound peaks they had identiﬁed were signiﬁcantly enriched at loci of genes in the“Wnt receptor signaling pathway”gene ontology(GO)term and on this basis proposed that hTR in com-plex with hTERT cooccupies Wnt target genes(31).Given the evidence described above that a previously reported interaction between hTERT and␤-catenin protein can be explained by cross-reactivity of anti-FLAG antibody rather than a bonaﬁde interac-tion,we used a bioinformatics approach to reexamine any poten-tial connection between the published genomic localizations of BRG1cross-linked sites,telomerase RNA cross-linked sites,and Wnt signaling genes.To identify loci on the HeLa S3genome enriched for localization sites of both BRG1and hTR(with hTR inferred to likely be in complex with hTERT,as was described previously[31]),we merged the published BRG1-enriched local-ization sites identiﬁed by ChIP-seq in HeLa S3cells(32)and the published hTR-enriched localization sites(31).We applied the criterion that they were separated byՅ100bp,as the same crite-rion was previously used to determine cooccupancy of BRG1with other members of the SWI/SNF complex at genomic loci(32). Using this criterion,217genes in the vicinity of the merged BRG1/ hTR-enriched loci were identiﬁed.However,while“Positive reg-ulation of apoptosis”was identiﬁed as a highly signiﬁcant GO term among these217genes,Wnt signaling was not identiﬁed as a signiﬁcant GO term(Table1).Of the217genes,only MYC is known to be a target gene of Wnt signaling.Thus,this analysis in HeLa S3cells(applying the criterion of peak separation no greater than100bp)failed to verify any signiﬁcant cooccupancy by hTR and BRG1of Wnt target genes or their nearby controlling regionsFIG3hTERT does not interact with␤-catenin or BRG1.(A)Anti-␤-catenin antibody cross-reacts with anti-FLAG immunoprecipitate.HeLa cells were either transfected with pcDNA3-FLAG-hTERT for16h(left)or left untransfected(right),followed by treatment with25mM LiCl or no LiCl treatment.Precipitates from anti-␤-catenin(clone14)or anti-FLAG(M2antibody F3165,Sigma)IP were subjected to Western blotting(WB)with anti-␤-catenin(clone14)or anti-FLAG antibodies.The asterisk indicates a slower-migrating background band.(B)Top:Western blot of HeLa cells transfected with FLAG-hTERT(right hand side)or left untransfected(left hand side)following IP with speciﬁc antibodies.M indicates size marker lane.Bottom:Western blot of input samples from experiment.(C)Top:Western blot of HeLa cells treated with or without␤-catenin shRNA following IP with speciﬁc antibodies.Bottom:Western blot of input samples from the experiment.Listerman et al. Molecular and Cellular Biologyexcept for MYC.This analysis does not directly address hTERT and BRG1cooccupancy on chromatin genomic loci.However, because some fractions of hTERT and hTR exist as telomerase complexes in HeLa S3cells,colocalization of hTR with BRG1at Wnt signaling gene loci might be predicted if hTERT protein and BRG1protein interact,as suggested previously(31).Our negative ﬁnding for hTR and BRG1cooccupancy at Wnt signaling gene loci thus does not support,but by itself does not refute,the possibility of interaction between hTERT and BRG1.Effects of hTERT overexpression on Wnt signaling target gene expression in cell lines do not reﬂect cellular Wnt signaling competency.While our bioinformatics data analysis did not iden-tify a signiﬁcant overlap of Wnt pathway genes with genomic loci cross-linkable to hTR and BRG1in HeLa cells,this analysis did not exclude the possibility that TERT,possibly not bound to hTR,still promotes Wnt signaling.To test directly whether hTERT modu-lates endogenous Wnt signaling target gene expression in breast cancer cells,we stably overexpressed hTERT in the high-Wnt-signaling SUM149PT and HCC3153cells and,as a control,in the Wnt-signaling-impaired HCC1806cells.We then measured the mRNA levels of84endogenous Wnt downstream target genes supplied as arrays for quantitative RT-PCR(qRT-PCR)(see Ma-terials and Methods).First,with hTERT overexpression(Fig.4C), the Wnt-signaling-competent SUM149PT cells showed a modest overall trend to greater expression of the Wnt target genes as a group compared with vector controls;however,this trend was no greater than that seen for the Wnt-signaling-impaired HCC1806TABLE1Signiﬁcant GO terms for genes close to BRG1and hTR co-occupied loci in HeLa S3genomeGene GO term P valueGO:0043065Positive regulation of apoptosis0.005GO:0006928Cell motion0.025GO:0051693Actinﬁlament capping0.027GO:0007028Cytoplasm organization0.035GO:0006917Induction of apoptosis0.035GO:0048146Positive regulation ofﬁbroblast proliferation0.037GO:0030834Regulation of actinﬁlament depolymerization0.040GO:0033043Regulation of organelle organization0.043GO:0032272Negative regulation of protein polymerization0.045GO:0032271Regulation of protein polymerization0.046FIG4Effect of hTERT overexpression on Wnt target gene expression in breast cancer cell lines.SABiosciences qPCR arrays were used to measure the effect of hTERT overexpression on the mRNA expression of endogenous Wnt target genes after treatment with25mM LiCl for6h.(A)Scatter plots of log-transformed relative expression levels of each gene.Red lines indicate a2.5-fold change in gene expression.(B)Wnt target genes that changedϮ2.5-fold compared to the control results(indicated by dotted line)upon hTERT overexpression in SUM149PT,HCC1806,and HCC3153cells.(C)hTERT mRNA expression relative to GAPDH mRNA and vector control expression in SUM149PT,HCC1806,and HCC3153.Bars represent means of the results of3(SUM149PT and HCC1806)and 2(HCC3153)biological replicates.Error bars indicate standard deviations(SD).hTERT Effects on Wnt Signaling in Breast Cancer 285。

药学英语试题及答案

药学英语试题及答案一、选择题（每题2分，共20分）1. The term "pharmacology" refers to the study of:A. The origin of drugsB. The effects of drugs on the bodyC. The synthesis of drugsD. The distribution of drugs答案：B2. Which of the following is not a route of drug administration?A. OralB. IntravenousC. InhalationD. Electrolysis答案：D3. The half-life of a drug is the time it takes for the concentration of the drug in the body to:A. DoubleB. TripleC. QuadrupleD. Decrease by half答案：D4. Which of the following is a common side effect of antibiotics?A. Dry mouthB. DiarrheaC. InsomniaD. All of the above答案：B5. The abbreviation "IV" stands for:A. IntravenousB. IntramuscularC. IntraperitonealD. Intradermal答案：A6. The term "bioavailability" refers to the:A. Percentage of a drug that is absorbed into the systemic circulationB. Percentage of a drug that is excreted unchangedC. Percentage of a drug that is metabolized in the liverD. Percentage of a drug that is stored in fat tissues答案：A7. Which of the following is a type of drug interaction?A. SynergismB. AntagonismC. PotentiationD. All of the above答案：D8. The therapeutic index of a drug is a measure of its:A. EfficacyB. SafetyC. Cost-effectivenessD. Taste答案：B9. The term "prodrug" refers to a drug that:A. Is already active when administeredB. Requires metabolic activation to become activeC. Is a combination of two drugsD. Is a drug that has been discontinued答案：B10. Which of the following is a method for enhancing drug solubility?A. Salt formationB. Coating with a polymerC. MicronizationD. All of the above答案：D二、填空题（每空1分，共20分）1. The ________ of a drug refers to its ability to reach the site of action in the body.答案：pharmacokinetics2. A drug that is administered as a liquid and is intended to be swallowed is called a ________.答案：solution3. The ________ of a drug is the maximum amount that can be given without causing harmful effects.答案：therapeutic dose4. A drug that is used to treat a specific disease or condition is called a ________.答案：therapeutic agent5. The ________ of a drug is the minimum amount that will produce a therapeutic effect.答案：therapeutic dose6. A drug that is used to prevent a disease or condition is called a ________.答案：prophylactic agent7. The ________ of a drug is the study of its effects on biological systems.答案：pharmacodynamics8. A drug that is used to alleviate symptoms without treating the underlying cause is called a ________.答案：symptomatic agent9. The ________ of a drug is the process by which it isremoved from the body.答案：elimination10. A drug that is used to treat a wide range of conditionsis called a ________.答案：broad-spectrum agent三、简答题（每题10分，共40分）1. Explain the difference between a generic drug and a brand-name drug.答案：A generic drug is a copy of a brand-name drug that has the same dosage form, safety, strength, quality, performance characteristics, and intended use. A brand-name drug is the original version of a drug that has beendeveloped by a pharmaceutical company and is protected by a patent.2. What are the factors that can influence the absorption ofa drug?答案：Factors that can influence the absorption of a drug include the route of administration, the formulation of the drug, the presence of food in the stomach, the pH of the gastrointestinal tract, and the individual's health status.3. Describe the process of drug metabolism.答案：Drug metabolism is the process by which the body breaks down and eliminates drugs. It typically involves two phases: Phase I reactions, which involve oxidation, reduction, or hydrolysis to make the drug more polar, and Phase。

经合组织风险认识工具OECD Risk Awareness tool

OECD Risk Awareness Tool for Multinational Enterprises in Weak
Governance Zones
ORGANISATION FOR ECONOMIC CO-OPERATION AND DEVELOPMENT
ORGANISATION FOR ECONOMIC CO-OPERATION AND DEVELOPMENT

Foreword
The OECD Risk Awareness Tool for Multinational Enterprises in Weak
Governance Zones aims to help companies that invest in countries where governments are unwilling or unable to assume their responsibilities. It addresses risks and ethical dilemmas that companies are likely to face in such weak governance zones, including obeying the law and observing international instruments, heightened care in managing investments, knowing business partners and clients and dealing with public sector officials, and speaking out about wrongdoing.
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靳智的英文翻译

模拟工业湿法烟气脱硫的仿真操作系统L.E. Kallinikos a, E.I. Farsari b, D.N. Spartinos b, N.G. Papayannakos a 摘要:这项工作中，由发电厂提出的一种模拟喷雾塔的湿法烟气脱硫系统简称（FGD），目的在于实现一个有效的后续行动和烟气脱硫操作系统的运行优化。

这种仍在完善的动态模型在相当长的时间内仍将收集运行数据来对模拟系统的性能进行验证，仿真模型也考虑了过程中的所有物理和化学过程, 如石灰石的分解，亚硫酸钙和石膏和亚硫酸盐离子的氧化结晶，然而气体被液滴吸收则是基于双膜理论。

对于影响泥浆液滴的平均直径的系统性能也进行了检查，因为它是作为操作系统正常运行的指标系数。

在模型不断发展的基础之上，人们也发现了该系统的局限性，得出的结论是该模拟系统对于不同的二氧化硫负荷脱硫效率有显著的区别，同时对二氧化硫的吸收率受到塔内液体的分散均匀程度的强烈影响。

关键词:气脱硫化;传质;面面积;态模型1前言在发电厂，含有硫磺的化石燃料被喂入燃烧室内燃烧生成二氧化硫气体，这种气体对于人类的生存环境和生活质量产生严重的影响。

因此，全球地区对于发电厂排放的气体中二氧化硫的含量已经制定了非常严格的规格标准。

欧洲共同体已经通过了两项指令，以减少化石燃料的发电厂排放的废气。

已开发的几种烟气脱硫方法包括湿式净化器，喷雾干燥塔，干燥净化器, 干洗涤器，再循环系统以及结合SO2/Nox的清除过程。

最常见的过程中使用的是全球湿法脱硫方法用石灰石作吸收剂，主要是就有吸二氧化硫的高效率（大于90％）。

此外，廉价的石灰石反应物的使用和生产的石膏（(CaSO4·2H2O)）都是这一进程的显着优点。

湿法脱硫的组成单元中二氧化硫的吸收是在吸收塔内完成的，吸收塔有好几种类型，如喷雾洗涤塔，填料塔，喷射鼓泡反应器，双回路塔。

工业实践中最常用的是喷雾塔吸收塔。

在这种类型的吸收塔中，由处在较高位置的三到五层喷嘴喷入石灰石浆，同时而烟道气体流量从底部逆流模式向塔的顶端与石灰石浆液液滴相遇发生中和反应。

拟胆碱药课件

药物毒扁豆碱吡斯的明依酚氯铵
安贝氯铵
加兰他敏
地美溴铵
特点
用途
脂溶性高
青光眼、中麻催醒
慢而持久
重症肌无力
快、短
重症肌无力的诊断和鉴别诊
断。
持久
不能耐受新斯的明或吡斯的
明的重症肌无力者。
弱，脂溶性好重症肌无力、脊髓灰白质炎
后遗症、非去极化肌松药中
毒、阿茨海默病。
易透过角膜，青光眼
作用持久
常见有机磷酸酯类化合物 Back
Section 1 Cholinoceptor agonists
一、M 、N 受体激动药 (choline esters)
（一）代表药
Acetylcholine（乙酰胆碱，Ach）
【physico-chemical property】
不稳定，极易被AchE水解而失活，故作用短暂。加之其作用广泛，故无临床应用价值，仅作工具药使用。Ach 难入中枢，故无明显中枢作用。
Mechanism of poisoning： Fig.
Manifestation of poisoning
Cholinergic Agents 12
轻：M样症状
Tab.
中：M样症状+N样症状
重：M样症状+N样症状+中枢症状
【prevention and cure】
1、预防：安全教育
2、诊断：接触史、AchE活性检查
4. Which appearance are there in Organophosphate poisoning? And how do you
treat it？
To prepare lessons for next class

小议“hm^2”

小议“hm^2”
孙二虎;张浙川
【期刊名称】《编辑学报》
【年(卷),期】2000(12)2
【摘要】hm2 是可与SI单位并用的我国法定计量单位 ;ha并不是“国际符号”或“国际通用符号” ,而是国家计量单位委员会认可暂时使用的单位。

我国学术界特别是编辑界对hm2 尚未完全接受 ,使用中有混乱现象。

1993年确立了hm2 法律地位的最新标准中反倒将“ha”定为“国际通用的”符号 ,建议有关部门对此作出恰当的修改。

【总页数】2页(P108-109)
【关键词】hm^2;公顷hA;法定计量单位;编辑工作;标准化
【作者】孙二虎;张浙川
【作者单位】<应用与环境生物学报>编辑部;<天然产物研究与开发>编辑部【正文语种】中文
【中图分类】TB91;G232
【相关文献】
1.In Silico Investigation of Agonist Activity of a Structurally Diverse Set of Drugs to hPXR Using HM-BSM and HM-PNN [J], Yi-ming ZHANG;Mei-jia CHANG;Xu-shu YANG;Xiao HAN
2.春小麦6750kg/hm2～7500kg/hm2高产栽培技术 [J], 桑加拉;欧图海
3.HM46,HM56和HM68优质液压油的研制及应用 [J], 颜贤忠
4.含氮有机物在Pt/HM,Pd/HM和CuO/HM催化剂上的氧化降解 [J], 刘泽菊;朱波;钟依均;罗孟飞
5.农业部：我国2016年试点耕地轮作33．33万hm2,休耕7．73万hm2 [J],因版权原因，仅展示原文概要，查看原文内容请购买。

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