Effects of Molybdenum on the Intermediates of Chlorophyll Biosynthesis in Winter Wheat Cultivars
牡蛎寡肽对免疫低下小鼠模型免疫功能的影响
牡蛎寡肽对免疫低下小鼠模型免疫功能的影响刘淑集;蔡水淋;刘智禹;许旻;苏永昌;王茵;潘南;乔琨;吴靖娜;苏捷;张伯超【期刊名称】《华南师范大学学报（自然科学版）》【年(卷),期】2018(050)002【摘要】The effect of oyster oligopeptide on regulation of immune function in cyclophosphamide （CTX)-induced immunosuppressive mice was evaluated.The indexes of immune organs such as liver and spleen,the transformation and propagation ratio of T lymphocyte,the activity of NK cell,the humoral immunity,macrophage phagocytosis,the level of cell cytokines and hemolysin were assessed.The experiment results revealed that compared with the model immunosuppressed group,oyster oligopeptide could significantly enhance the proliferation of spleen lymphocyte,NK cell activity,expurgation index k,devouring index a,ability of devouring neutral red,the level of TNF-α、IL-6 and hemolysin,and regulate the distribution of CD3+ 4T and CD3+ 8T spleen lymphocytesubsets.However,the influence of oyster oligopeptide on the immune organs was not significant.It was concluded that oyster oligopeptide can improve the cell immunity,humoral immunity and non-specific immune function of immunosuppressive mice induced by CTX,and it can regulate the immune function positively in mice.%采用腹腔注射环磷酰胺(Cyclophosvnamide,CTX)建立免疫低下小鼠模型,研究不同剂量牡蛎寡肽对小鼠免疫器官脏器指数、脾淋巴细胞转化增殖情况、自然杀伤细胞(Natural Killercell,NK)细胞活性、小鼠体液免疫、巨噬细胞吞噬能力、血清TNF-α、IL-6和溶血素水平的影响.结果显示:与免疫抑制模型组小鼠相比,牡蛎寡肽能够显著提高脾淋巴细胞增殖能力、NK细胞活性、廓清指数K、吞噬指数a、吞噬中性红能力、TNF-α、IL-6、溶血素水平和脾淋巴细胞CD3+ 4T淋巴细胞亚群及CD3+ 8T淋巴细胞亚群分布(P＜0.05),而对小鼠的肝、脾脏指数影响不显著,表明牡蛎寡肽能够提高由CTX引起的免疫低下模型小鼠的细胞免疫、体液免疫及非特异性免疫功能,对小鼠的免疫功能具有正面调控的作用.【总页数】7页(P70-76)【作者】刘淑集;蔡水淋;刘智禹;许旻;苏永昌;王茵;潘南;乔琨;吴靖娜;苏捷;张伯超【作者单位】福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013;福建农林大学食品科学学院,福州350002;福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013;福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013;福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013;福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013;福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013;福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013;福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013;福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013;福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013;福建省水产研究所∥福建省海洋生物增养殖与高值化利用重点实验室∥福建省海洋生物资源开发利用协同创新中心,厦门361013【正文语种】中文【中图分类】R151【相关文献】1.槲皮素对免疫低下小鼠免疫功能的影响 [J], 田瑞雪; 孙耀宗; 姚有昊; 张子健; 张继民; 宋维芳2.红景天当归不同配伍对免疫低下小鼠免疫功能的影响 [J], 史顶聪; 赵宏宇; 佟晓乐; 张凤清3.鹰嘴豆肽对免疫低下小鼠免疫功能的影响 [J], 李睿珺;秦勇;周雅琳;刘伟;李雍;于兰兰;陈宇涵;许雅君4.中药复方“益智灵”对臭氧所致衰老免疫低下小鼠模型免疫功能改善作用研究[J], 周勇;严宣佐;徐秋萍;吴金英5.绞股蓝总苷对免疫功能低下小鼠模型特异性免疫功能的影响 [J], 周俐;叶开和;任先达因版权原因，仅展示原文概要，查看原文内容请购买。
莫诺苷对局灶性脑缺血再灌注大鼠皮层IL-1β的影响
Ｋｅ ｏ ｄ ：ｍｏ ｒ ｎ ｓｄ ；ｃ ｒ ｂ ａ ｓ ｈ ｍｉ／ ｅ ｅ ｆ ｓｏ ；Ｉ 一 Ｂ ｎ ｉ ｆ ｍｍａ ｏ ｙ ｅ ｒ ｐ ｏ ｅ ｔ ｅ ｙｗ ｒ ｓ ｒ ｏ ｉｉ ｅ ｅ ｅ ｒ ｌｉ ｅ ａ ｒ ｐ ｒｕ ｉ ｎ Ｌ １ ；ａ ｔｉ ｌ ｃ ｎ ａ ｔ ｒ ；ｎ ｕ ０ ｒ ｔ ｃ ｉ ｖ
Ｈｏｐｉ ｌｏ ａ ｔ ｌ ｅ ｉａ ｎ ｖ ｒｉ ｓ ｔ ｆ Ｃ ｐｉ ｄ ｃ ｌＵ ｉｅｓｔ ａ ａ Ｍ ｙ，Ｂｅｊ ｎ ０ ０ ３ ｈ ｎ ｉｉｇ １ ０ ５ ，Ｃ ｉａ Ａｂｔａ ｔ ｓｒ ｃ：Ｏｂｅ ｔ ｅ Ｔｏｉｖ ｓｉａｅｔ ｅｅｆｃｓｏ ｒｏ ｉｉｅｏ Ｌ １ ｎ ｒｔｃ ｒｅ ｔ ｏ ａ ｃ ｒｂ ａ ｓ ｈ ｍｉ／ ｅ ｅ ｆｓｏ ． ｊｃｉ ｎ ｅ ｔ ｔ ｈ ｆｅｔ ｆｍｏ ｒｎｓｄ ｎ Ｉ 一ｐ ｉ ａ ｏ ｔｘ ｗｉ ｆｃ ｌ ｅｅ ｒ ｌｉｃ ｅ ａ ｒｐ ｒｕ ｉｎ ｖ ｇ ｈ
艾厚 喜 汪 莹 ， 栋 明。 王 文 ， 东 明 ， 丽 ， 林 ， 许 ， 高 张 李
［ 要］ 目 的 研 究 山茱 萸有 效 成 分 莫 诺 苷对 局 灶 性 脑 缺血 再 灌 注 大 鼠皮 层 Ｉ 一８的影 响 。方 法 用 线 栓法 制 备 大 鼠局 灶性 脑 摘 Ｉｌ
缺 血 再 灌 注模 型 ， 缺血 ３ ｎ， 灌 注 ７ ０ ｍｉ 再 ２ｈ。 以秋 水 仙 碱 为 阳 性 对 照 药 ， 用 酶 联 免 疫 吸 附 法 （ ＩＩ Ａ） 测 大 鼠脑 皮 层 炎 症 因子 采 Ｅ ｓ 检 Ｉ ｔ Ｄｌ ３的变 化 。结果 奠 诺 苷 可 剂 量依 赖 性 地 明 显 降低 由脑 缺 血 再灌 引起 的 Ｉ Ｂ含 量 增 加 （ Ｌ１ Ｐ＜ ｏ ０ １ 。结 论 莫 诺 苷 能 通 过 增 强 ．０ ） 大 鼠皮层 抗 炎 能 力 发挥 神 经 保 护 作 用 。
基于网络药理学探讨白藜芦醇治疗肺癌的生物分子机制
基于网络药理学探讨白藜芦醇治疗肺癌的生物分子机制 张丽慧，耿其顺，朱子家，王文斌，沈志博，李砺锋，薛文华，赵杰郑州大学第一附属医院，河南郑州 450052摘要：目的 基于网络药理学，运用在线数据库研究白藜芦醇治疗肺癌的潜在作用机制。
方法 通过中药系统药理学数据库与分析平台（TCMSP）、PubChem、SwissTargetPrediction、GeneCards数据库分别获取白藜芦醇和肺癌的相关基因，取二者交集基因，使用String数据库获得交集基因的蛋白相互作用（PPI）网络图，使用DA VID6.8对交集基因进行GO和KEGG富集分析。
采用Cytoscape3.7.2软件构建化合物-靶点-通路-疾病网络模型。
使用AutoDock4.2.6软件对白藜芦醇和重要靶点进行分子对接。
Western blot检测白藜芦醇对人肺癌H1975细胞p-Akt蛋白表达的影响。
结果 获得白藜芦醇和肺癌交集基因78个，PPI网络图表明交集基因关系密切。
富集分析得到生物过程55项、分子功能26项、细胞组分18项，以及86条相关通路，其中以PI3K-Akt 通路富集靶点较多。
分子对接结果显示，白藜芦醇与PIK3CB、PIK3CA这2个重要靶点均能稳定结合。
Western blot检测结果显示，白藜芦醇能够显著降低人肺癌H1975细胞p-Akt蛋白表达。
结论 白藜芦醇可能主要通过作用于PIK3CB、PIK3CA靶点，介导PI3K-Akt信号通路发挥治疗肺癌作用。
关键词：白藜芦醇；肺癌；网络药理学；作用机制；分子对接中图分类号：R273.42；R285 文献标识码：A 文章编号：1005-5304(2021)06-0046-06DOI：10.19879/ki.1005-5304.202004113 开放科学（资源服务）标识码（OSID）：Biomolecular Mechanism of Resveratrol Against Lung Cancer Based on Network Pharmacology ZHANG Lihui, GENG Qishun, ZHU Zijia, WANG Wenbin, SHEN Zhibo, LI Lifeng, XUE Wenhua, ZHAO Jie First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China Abstract:Objective To use the online databases to study the potential mechanism of resveratrol in treating lung cancer based on network pharmacology. Methods The genes related to resveratrol and lung cancer were obtained from TCMSP, PubChem, SwissTargetPrediction and GeneCards database, and the related intersection genes of resveratrol and lung cancer were screened out. The String database was used to draw the PPI network diagram of the intersection genes, and the DA VID 6.8 was used to perform GO and KEGG enrichment analysis. The compound-target-signal pathway-disease network model was constructed by using Cytoscape 3.7.2 software. AutoDock 4.2.6 software was used to perform molecular docking of resveratrol and important targets. Western blot was used to detect the effect of resveratrol on the expression of p-Akt protein in human lung cancer H1975 cell line. Results Totally 78 intersection genes of resveratrol and lung cancer were obtained, and the PPI network diagram showed that there was a close relationship between them. The enrichment analysis mainly obtained 55 biological processes, 26 molecular functions, 18 cellular components, and 86 related pathways, among which the PI3K-Akt pathway enriched the most targets. Molecular docking results showed that resveratrol could stably bind to two important targets, PIK3CB and PIK3CA. Western blot test results showed that resveratrol could significantly reduce the expression of p-Akt protein in human lung cancer H1975 cells. Conclusion Resveratrol may mainly act on the targets of PIK3CB and PIK3CA, and mediates the PI3K-Akt signaling pathway to exert anti-lung cancer action.Keywords: resveratrol; lung cancer; network pharmacology; action mechanism; molecular docking研究表明，肿瘤的发生发展与基因突变及多条信号通路改变有关[1-2]。
白念珠菌群体感应分子法尼醇对巨噬细胞Ana-1和RAW264.7增殖、凋亡和迁移能力的影响
doi：10.3969/j.issn.1000-484X.2021.09.004白念珠菌群体感应分子法尼醇对巨噬细胞Ana-1和RAW264.7增殖、凋亡和迁移能力的影响①胡万超王丹朱俊峰周子阳唐建国（复旦大学附属上海市第五人民医院急危重症医学科，上海200240）中图分类号R392.5文献标志码A文章编号1000-484X（2021）09-1041-06［摘要］目的：探讨白念珠菌群体感应分子法尼醇对巨噬细胞Ana-1和RAW264.7增殖、凋亡和迁移能力的影响。
方法：实验分为Ana-1细胞对照组、RAW264.7细胞对照组、不同浓度法尼醇+Ana-1细胞组和不同浓度法尼醇+RAW264.7细胞组。
对照组给予无血清培养基及1‰二甲基亚砜（DMSO），法尼醇+Ana-1细胞组和法尼醇+RAW264.7细胞组给予无血清培养基配制的各半数抑制浓度（IC50）及不同浓度法尼醇。
CCK-8法检测各组巨噬细胞增殖抑制率，Annexin V-FITc法检测各组巨噬细胞凋亡率，Western blot检测凋亡相关蛋白表达，Transwell小室检测巨噬细胞迁移能力。
结果：与对照组相比，法尼醇处理组对巨噬细胞Ana-1和RAW264.7增殖的抑制作用显著增强，且随浓度增加，抑制效果越明显（P<0.05）；法尼醇处理巨噬细胞组细胞凋亡率显著升高，且呈浓度依赖性（P<0.05），法尼醇可能通过影响磷酸化AKT、Bcl-2、Cleaved Caspase-3、Cleaved Caspase-9及Cleaved PARP表达促进RAW264.7和Ana-1巨噬细胞凋亡；Transwell结果显示随法尼醇浓度增加，巨噬细胞迁移数增加。
结论：白念珠菌群体感应分子法尼醇可有效抑制巨噬细胞Ana-1和RAW264.7增殖、促进其凋亡和迁移，其机制可能与降低磷酸化AKT、Bcl-2、PARP，上调Cleaved Caspase-3，Cleaved Caspase-9和Cleaved PARP表达有关。
新教材同步备课2024春高中生物第3章基因的本质3.3DNA的复制课件新人教版必修2
(2)注意碱基的单位是“对”还是“个”。 (3)切记在DNA复制过程中，无论复制了几次，含有亲代脱氧 核苷酸单链的DNA分子都只有两个。 (4)看清试题中问的是“DNA分子数”还是“链数”，“含” 还是“只含”等关键词，以免掉进陷阱。
二、DNA分子的复制
例1.某DNA分子中含有1 000个碱基对(被32P标记)，其中有胸腺 嘧啶400个。若将该DNA分子放在只含被31P标记的脱氧核苷酸的 培养液中让其复制两次，子代DNA分子相对分子质量平均比原来 减少 1 500 。
F2:
提出DNA离心
高密度带 低密度带 高密度带
低密度带 高密度带
一、DNA复制的推测—— 假说-演绎法
1.提出问题 2.提出假说
（1）演绎推理 ③分散复制
15N 15N
提出DNA离心
P:
3.验证假说
15N 14N
F1:
细胞分 裂一次
转移到含 14NH4Cl的培养 液中
提出DNA离心
细胞再 分裂一次
二、DNA分子的复制
例3.若亲代DNA分子经过诱变，某位点上一个正常碱基变成了5-溴 尿嘧啶（BU），诱变后的DNA分子连续进行2次复制，得到4个子 代DNA分子如图所示，则BU替换的碱基可能是（ C ）
A.腺嘌呤 C.胞嘧啶
B.胸腺嘧啶或腺嘌呤 D.鸟嘌呤或胞嘧啶
二、DNA分子的复制
例4. 5-BrU(5-溴尿嘧啶)既可以与A配对，又可以与C配对。将一 个正常的具有分裂能力的细胞，接种到含有A、G、C、T、5-BrU 五种核苷酸的适宜培养基上，至少需要经过几次复制后，才能实现 细胞中某DNA分子某位点上碱基对从T—A到G—C的替换（ B ）
去甲基化和去乙酰化
Repression of induced apoptosis in the 2-cell bovine embryo involves DNA methylation and histone deacetylationSilvia F.Carambula,Lilian J.Oliveira,Peter J.Hansen *Department of Animal Sciences,University of Florida,P.O.Box 110910,Gainesville,FL 32611-0910,USAa r t i c l e i n f o Article history:Received 2August 2009Available online 8August 2009Keywords:ApoptosisPreimplantation embryo DNA methylation Histone acetylation 5-Aza-20-deoxycytidine Trichostatin Aa b s t r a c tApoptosis in the bovine embryo cannot be induced by activators of the extrinsic apoptosis pathway until the 8–16-cell stage.Depolarization of mitochondria with the decoupling agent carbonyl cyanide 3-chlo-rophenylhydrazone (CCCP)can activate caspase-3in 2-cell embryos but DNA fragmentation does not occur.Here we hypothesized that the repression of apoptosis is caused by methylation of DNA andTUNEL was affected by a treatment ÂCCCP interaction (P <0.0001).CCCP did not cause a large increase in the percent of cells positive for TUNEL in embryos treated with vehicle but did increase the percent of cells that were TUNEL positive if embryos were pretreated with AZA or TSA.Immunostaining using an antibody against 5-methyl-cytosine antibody revealed that AZA and TSA reduced DNA methylation.In conclusion,disruption of DNA methylation and histone deacetylation removes the block to apoptosis in bovine 2-cell embryos.Ó2009Elsevier Inc.All rights reserved.IntroductionDuring preimplantation development,the mammalian embryo goes through a period where it is resistant to proapoptotic signals.In the best studied example,the bovine,this period lasts from the 2-cell stage through the 8–16-cell stage [1–5].Inhibition of the extrinsic pathway for apoptosis at the 2-cell stage is caused in part by resistance of the mitochondria to depolarization [4,5].In addi-tion,a second block exists that is revealed when the mitochondrial membrane is artificially depolarized by carbonyl cyanide 3-chloro-phenylhydrazone (CCCP).In this case,caspase-9and caspase-3activation takes place but DNA fragmentation does not occur [4].Thus,DNA is resistant to caspase-3mediated events such as activa-tion of caspase-activated DNase (CAD).One possible explanation for DNA resistance to CAD may reside with the structure of DNA in the early preimplantation embryo.At the 2-cell stage,little transcription takes place [6–7]and DNA is highly methylated [8].DNA demethylation occurs over the next several cleavage divisions [9].Thus,the stage of development at which susceptibility to apoptosis is acquired (the 8–16-cell stage)is also a time of when DNA methylation is reduced [9]and tran-scription is activated [6].DNA methylation can reduce the accessibility of DNases to DNA as shown for DNase I [10].Here we hypothesize that the repression of apoptosis responses in response to mitochondrial depolarization in the 2-cell embryo is caused by DNA methylation that makes internucleosomal DNA inaccessible to activated CAD.Moreover,we hypothesize that repression requires deacetylated histones.Materials and methodsReagents.Materials for in vitro maturation of oocytes,in vitro fer-tilization,and embryo culture were obtained as described previously [11].Carbonyl cyanide 3-chlorophenylhydrazone (CCCP)was pur-chased from Sigma (St.Louis,MO)and was maintained in 100mM stocks in dimethyl sulfoxide (DMSO)at À20°C in the dark.The CCCP stock solution was diluted in embryo culture medium (called KSOM-BE2,see Ref.[12]for recipe)to 100l M in 0.1%DMSO on the day of use.5-Aza-20-deoxycytidine (AZA)and trichostatin-A (TSA)were ob-tained from Sigma and used at a final concentration of 100l M and 100nM,respectively.The In Situ Cell Death Detection Kit (TMR red)was from Roche Diagnostics Corporation (Indianapolis,IN),Hoescht 33342was from Sigma,polyvinylpyrrolidone (PVP)was from Eastman Kodak (Rochester,NY).Anti-5-methylcytosine (mouse IgG1;clone 16233D3)was purchased from Calbiochem (San Diego,CA).The Zenon Alexa Fluor 488mouse IgG1labeling kit 488and Prolong ÒAntifade Kit were obtained from Invitrogen0006-291X/$-see front matter Ó2009Elsevier Inc.All rights reserved.doi:10.1016/j.bbrc.2009.08.029*Corresponding author.Fax:+13523925595.E-mail address:Hansen@animal.ufl.edu (P.J.Hansen).Biochemical and Biophysical Research Communications 388(2009)418–421Contents lists available at ScienceDirectBiochemical and Biophysical Research Communicationsjournal homepage:www.else v i e r.c o m /l o ca t e /y b b r cMolecular Probes(Eugene,OR).All other reagents were purchased from Sigma or Fisher Scientific(Pittsburgh,PA).Experiment1—Effects of cytosine demethylation and inhibition of histone deacetylation on induction of apoptosis by CCCP in2-cell em-bryos.Procedures for production of embryos in vitro were per-formed as previously described[12].After fertilization of matured oocytes for8h at38.5°C in an atmosphere of5%(v/v) CO2in humidified air,putative zygotes were cultured in groups of30in50-l l microdrops of KSOM-BE2overlaid with mineral oil at38.5°C in a humidified atmosphere of5%(v/v)CO2and5%(v/ v)O2with the balance N2.At18h post insemination(hpi),embryos were harvested and placed in groups of30in fresh50-l l micro-drops of KSOM-BE2containing either0.1%DMSO(vehicle), 100l M AZA or100nM TSA.At28–30hpi,2-cell embryos were harvested and placed in groups of10–20in50-l l microdrops of KSOM-BE2containing the same treatment as previously(vehicle, AZA or TSA)and either vehicle(0.1%DMSO,v/v)or100l M CCCP. Embryos were cultured for24h,harvested and then analyzed for TUNEL labeling.Procedures for TUNEL were performed as described previously [13].Slides were examined using a Zeiss Axioplan2epifluores-cence microscope(Zeiss,Gottingen,Germany)with Zeissfilter sets 02(DAPIfilter)and15(rhodaminefilter).Digital images for epi-fluorescence and for light microscopy using differential interfer-ence contrast were acquired using AxioVision software(Zeiss) and a high-resolution black and white Zeiss AxioCam MRm digital camera.Images were merged for presentation.The Hoescht stain-ing was digitally converted to green before merger.The experiment was replicated six times using a total of458 embryos.Experiment2—Effects of cytosine demethylation and inhibition of histone deacetylation on DNA methylation.The experiment was con-ducted as for Experiment1except embryos were examined for DNA methylation at the end of the experiment using immunocyto-chemistry with an antibody against5-methylcytosine.Unless otherwise stated,reactions were at room temperature and re-agents were diluted in phosphate-buffered saline(PBS;10mM KPO4,pH7.4containing0.9%(w/v)NaCl)containing1mg/ml pol-yvinylpyrrolidone(PVP).Embryos were washed in PBS–PVP,fixed in4%(w/v)paraformaldehyde,washed in PBS–PVP,permeabilized with0.3%(v/v)Triton X-100for30min,washed extensively in 0.05%Tween20and treated with3M HCl for30min at37°C.After neutralization with100mM Tris–HCl,pH8.5containing1mg/ml PVP,embryos were washed in0.05%(v/v)Tween20and non-spe-cific binding sites blocked by incubation in a blocking buffer con-sisting of PBS–PVP containing2%(w/v)bovine serum albumin overnight at4°C.The anti-5-methylcytosine antibody used for visualization of DNA methylation was labeled with Fab fragments against mouse IgG conjugated to Alexa Flour488(ZenonÒMouse Labeling IgG kits,Invitrogen Molecular Probes)as per manufacturer’s instruc-tions.An irrelevant mouse IgG1was similarly labeled as an isotype control.The labeled complex was diluted in blocking buffer at afi-nal concentration of5l g/ml primary antibody and embryos were incubated for1h at room temperature in the dark.After several washes in0.05%(v/v)Tween20in PBS-PVP,embryos were placed on slides and coverslips mounted using ProlongÒAntifade reagent (Invitrogen).Embryos were examined using a Zeiss Axioplan2epi-fluorescence microscope with Zeissfilter sets02(DAPIfilter)and 03(FITC).Intensity of methylation was subjectively scored for each embryo on a scale of0(no methylation)to3.A total of61embryos in two replicates were analyzed.Statistical analysis.Data were analyzed by least-squares analysis of variance using the General Linear Models procedure of the Sta-tistical Analysis System(SAS for Windows,version9.2,SAS Insti-tute,Inc.,Cary NC).Dependent variables for Experiment1,calculated on an embryo basis,were total cell number and percent of cells that were apoptotic(i.e.,TUNEL positive).Independent variables included pretreatments(vehicle,AZA or TSA),CCCP(yes vs.no)and replicate.The mathematical model included main ef-fects and all interactions.Replicate was considered random and other main effects were consideredfixed.F tests were calculated using error terms calculated from expected means squares.Differ-ences between individual means were determined using the pdiff procedure of SAS.The dependent variable for Experiment3was methylation score and the independent variable was treatment.ResultsExperiment1—Effects of cytosine demethylation and inhibition of histone deacetylation on induction of apoptosis by CCCP in2-cell embryosIn thefirst experiment,embryos were treated with either AZA or TSA at the zygote stage to block cytosine methylation or histone deacetylation and then treated with CCCP at the2-cell stage.Rep-resentative images of TUNEL labeling are shown in Fig.1A–F,least-squares means±SEM for total cell number are in Fig.1G and least-squares means±SEM for the percent of cells that were TUNEL-po-sitive are in Fig.1H.Embryo growth,as determined by total cell number at the end of the experiment,was reduced by AZA,and to a lesser extent,TSA (P<0.05)(Fig.1G).Regardless of pretreatment,CCCP induced cell-cycle arrest as determined by a reduction in cell number (P<0.001)(Fig.1G).As shown in Fig.1H,the percent of blastomeres positive for TUN-EL was affected by a treatmentÂCCCP interaction(P<0.0001). CCCP did not cause a large increase in the percent of cells positive for TUNEL in embryos treated with vehicle(2.0±3.4%vs.7.7±5.5%;compare Fig.1A with D)but did cause a large increase in the percent of cells that were positive for TUNEL for embryos pre-treated with AZA(5.4±2.9%vs.42.3±3.2%;compare Fig.1B,E)or TSA(17.1±2.8%vs.24.9±4.2%;compare Fig.1C,F).The magnitude of the TUNEL labeling after CCCP depolarization was less for TSA than AZA(P<0.01)(Fig.1H).The degree of TUNEL labeling in the absence of CCCP was great-er for embryos treated with TSA than for control embryos or em-bryos treated with AZA(P<0.01).A total of32%of TSA-treated embryos were P8cells,a stage when apoptosis is possible.In this subset of TSA-treated embryos,the proportion of cells that were TUNEL-positive was25.7±4.1%.In control embryos P8cells,only 1.3±4.6%of cells were TUNEL positive.Thus,some of the TSA-trea-ted embryos underwent apoptosis when developing past the8-cell stage.None of the AZA-treated embryos were>8cells.Further analysis of the effect of CCCP on TSA-treated embryos focused on the subset of embryos that were<8cells(i.e.,those that are ordi-narily not susceptible to apoptosis).In this subset,which repre-sents68%of the TSA-treated embryos,there was an increase in the percent of blastomeres that were TUNEL positive after CCCP treatment(10.0±4.2%vs.24.4±4.5%,P<0.025).Experiment2—Effects of cytosine demethylation and inhibition of histone deacetylation on DNA methylationAs determined by reactivity with an antibody to5-methylcyto-sine,treatment of putative zygotes with AZA or TSA reduced DNA methylation at52–54hpi(Fig.2).In control embryos treated with vehicle,nuclei reacted strongly with antibody against5-methyl-cytosine(Fig.2A).Immunoreaction product was greatly reduced in embryos treated with AZA(Fig.2B)and reduced to a lesser ex-tent for embryos treated with TSA(Fig.2C).The subjective score for degree of DNA methylation was greatest in control embryosS.F.Carambula et al./Biochemical and Biophysical Research Communications388(2009)418–421419(2.5±0.1),least in AZA-treated embryos (1.0±0.1),and intermedi-ate in TSA-treated embryos (1.9±0.1).Differences between each mean were significant (P <0.0001).DiscussionThe bovine preimplantation embryo undergoes a period from the 2-cell stage to 8–16-cell stage when it is resistant to activators of the extrinsic pathway for induction of apoptosis [1–5].The block to apoptosis involves resistance of the mitochondria to depolariza-tion and failure of caspase-3activation to lead to DNA fragmenta-tion [4,5].Here we show that the resistance of DNA to caspase-3mediated events is the result of inaccessibility of the DNA caused by a chromatin structure dependent upon DNA methylation and histone acetylation.5-Aza-20-deoxycytidine inhibits DNA methylation by incor-poration into DNA during replication and subsequent inhibition of DNA methyltransferases (DNMT)[14].Treatment of embryos with AZA reversed the block to apoptosis so that CCCP treat-ment caused DNA fragmentation.Experiments with AZA indi-cate that inhibition of apoptosis caused by mitochondrial depolarization involves DNA methylation preventing accessibil-ity of CAD to DNA.One can visualize two mechanisms by which methylated cytosines could prevent enzymatic cleavage of DNA.Methylated cytosines can repel certain proteins,for example transcription factors [15],and may also repel CAD.Alternatively,methylated cytosines can attract other proteins,such as the Sin3A histone deacetylase complex and a methyl-CpG binding protein called MeCP2that binds tightly to chro-mosomes [15].Fig.1.DNA demethylation and histone acetylation allows DNA fragmentation in 2-cell embryos after apoptosis triggered by mitochondria depolarization.Putative zygotes were treated with vehicle (DMSO),5-aza-20-deoxycytidine (AZA)or trichostatin-A (TSA);2-cell embryos were collected at 28–30h post insemination and exposed to 100l M CCCP.Total cell number and the percent of cells positive for the TUNEL reaction were determined 24h later.Representative images of embryos following the TUNEL procedure are shown in panels A–F.Nuclei were labeled with Hoechst 33342(digitally colored as green)and those that are TUNEL-positive nuclei are additionally labeled with TMR red (red).(G)Total cell number and (H)the percent of blastomeres that are TUNEL positive.Data are least-squares means ±SEM for embryos cultured in the absence (black bars)and presence (white bars)of CCCP.Cell number was affected by pretreatment (P <0.025),CCCP (P <0.001)and the interaction (P <0.001).Percent of blastomeres positive for TUNEL was affected by CCCP (P <0.025)and the interaction between pretreatment and CCCP (P <0.001).Bars with different superscripts differ (P <0.05orless).Fig.2.Reduction in DNA methylation caused by treatment of putative zygotes with 5-aza-20-deoxycytidine (AZA)and trichostatin-A (TSA).DNA methylation was observed by the immunolocalization of 5-methylcytosine in bovine embryos at 52–54h after insemination using an anti-5-methycytosine tagged with Alexa Fluor 488(green).(For interpretation of color mentioned in this figure the reader is referred to the web version of the article.)420S.F.Carambula et al./Biochemical and Biophysical Research Communications 388(2009)418–421The fact that embryos treated with TSA were also capable of undergoing DNA fragmentation in response to CCCP suggests that repression of apoptosis involves histone interactions with DNA controlled by histone deacetylation.Results with TSA were more complex to interpret than for AZA because more TSA-treated em-bryos not exposed to CCCP experienced TUNEL labeling than for AZA-treated embryos.This effect is due to an increase in TUNEL labeling among TSA-treated embryos that were8cells or greater. Unlike for AZA,which caused a large reduction in cell number, TSA reduced developmental competence only slightly and many TSA embryos reached the8–16-cell stage.In these more advanced embryos,TSA caused apoptosis in the absence of CCCP.Given the role of DNA methylation and histone deacetylation in repressing apoptosis in early stages of development,it is proposed that the acquisition of the capacity for apoptosis at the8–16-cell stage is dependent upon loss of DNA methylation or changes in his-tone acetylation.There are large species differences in the pattern of DNA methylation during early development with some species like the mouse experiencing a continual reduction in DNA methyl-ation until the blastocyst stage while other species like the pig and rabbit do not experience large scale demethylation during early development[16].In the cow,DNA methylation is reduced from the2-cell to8-cell stage and then increases by the16-cell stage [9,17].There are also changes in histone acetylation that occur dur-ing development with Histone H4K5and K12becoming deacety-lated at the one and2-cell stages,followed by reacetylation that reaches a maximum at the8-cell stage[17].Given the importance of DNA methylation and histone deacet-ylation for repressing apoptosis in early cleavage-stage embryos, it is possible that some types of embryonic death result from inad-equate DNA methylation or histone deacetylation.Patterns of DNA methylation during early development are clearly important for embryonic development because AZA caused a large reduction in embryo cell number.In conclusion,repression of apoptosis in the2-cell embryo in-volves inaccessibility of caspase-activated DNases to the DNA med-iated by a chromatin structure determined by DNA methylation and histone deacetylation.Future work should focus on the particular interactions between methylated cytosines and histones responsi-ble for this inaccessibility as well as the importance of aberrant chro-matin structure and premature apoptosis in embryonic death.AcknowledgmentsFunding was provided by the National Research Initiative Com-petitive Grants Program Grant No.2007-35203-18070from the U.S.Department of Agriculture Cooperative State Research,Educa-tion and Extension Service.Lilian Oliveira was supported by a CAPES/Fulbright Fellowship.The authors thank William Rembert for collecting ovaries;Marshall,Adam,and Alex Chernin and employees of Central Beef Packing Co.(Center Hill,FL)for provid-ing ovaries;and Scott A.Randell of Southeastern Semen Services (Wellborn,FL)for donating semen.None of the authors have a con-flict of interest.References[1]F.F.Paula-Lopes,P.J.Hansen,Heat-shock induced apoptosis in preimplantationbovine embryos is a developmentally-regulated phenomenon,Biol.Reprod.66 (2002)1169–1177.[2]C.E.Krininger III,S.H.Stephens,P.J.Hansen,Developmental changes ininhibitory effects of arsenic and heat shock on growth of preimplantation bovine embryos,Mol.Reprod.Dev.63(2002)335–340.[3]P.Soto,R.P.Natzke,P.J.Hansen,Actions of tumor necrosis factor-a on oocytematuration and embryonic development in cattle,Am.J.Reprod.Immunol.50 (2003)380–388.[4]A.M.Brad,K.E.M.Hendricks,P.J.Hansen,The block to apoptosis in bovine two-cell embryos involves inhibition of caspase-9activation and caspase-mediated DNA damage,Reproduction134(2007)789–797.[5]L.A.de Castro e Paula,P.J.Hansen,Ceramide inhibits development andcytokinesis and induces apoptosis in preimplantation bovine embryos,Mol.Reprod.Dev.75(2008)1063–1070.[6]R.E.Frei,G.A.Schultz,R.B.Church,Qualitative and quantitative changes inprotein synthesis occur at the8–16-cell stage of embryogenesis in the cow,J.Reprod.Fertil.86(1989)637–641.[7]D.R.Natale,G.M.Kidder,M.E.Westhusin, A.J.Watson,Assessment bydifferential display-RT-PCR of mRNA transcript transitions and alpha-amanitin sensitivity during bovine preattachment development,Mol.Reprod.Dev.55(2000)152–163.[8]J.S.Park,Y.S.Jeong,S.T.Shin,K.K.Lee,Y.K.Kang,Dynamic DNA methylationreprogramming:active demethylation and immediate remethylation in the male pronucleus of bovine zygotes,Dev.Dyn.236(2007)2523–2533.[9]W.Dean,F.Santos,M.Stojkovic,V.Zakhartchenko,J.Walter,E.Wolf,W.Reik,Conservation of methylation reprogramming in mammalian development: aberrant reprogramming in cloned embryos,A98 (2001)13734–13738.[10]S.Kochanek, D.Renz,W.Doerfler,Differences in the accessibility ofmethylated and unmethylated DNA to DNase I,Nucleic Acids Res.21(1993) 5843–5845.[11]K.E.Hendricks,L.Martins,P.J.Hansen,Consequences for the bovine embryo ofbeing derived from a spermatozoon subjected to post-ejaculatory aging and heat shock:development to the blastocyst stage and sex ratio,J.Reprod.Dev.55(2009)69–74.[12]P.Soto,R.P.Natzke,P.J.Hansen,Identification of possible mediators ofembryonic mortality caused by mastitis:actions of lipopolysaccharide, prostaglandin F2a,and the nitric oxide generator,sodium nitroprusside dihydrate,on oocyte maturation and embryonic development in cattle,Am.J.Reprod.Immunol.50(2003)263–272.[13]Z.Roth,P.J.Hansen,Involvement of apoptosis in disruption of developmentalcompetence of bovine oocytes by heat shock during maturation,Biol.Reprod.71(2004)1898–1906.[14]W.G.Zhu,G.A.Otterson,The interaction of histone deacetylase inhibitors andDNA methyltransferases inhibitors in the treatment of human cancer cells, Curr.Med.Chem.Anticancer Agents3(2003)187–199.[15]A.Bird,DNA methylation patterns and epigenetic memory,Genes Dev.16(2002)6–21.[16]H.Fulka,J.C.St.John,J.Fulka,P.Hozák,Chromatin in early mammalianembryos:achieving the pluripotent state,Differentiation76(2008)3–14. [17]W.E.Maalouf,R.Alberio,K.H.Campbell,Differential acetylation of histone H4lysine during development of in vitro fertilized,cloned and parthenogenetically activated bovine embryos,Epigenetics3(2008)199–209.S.F.Carambula et al./Biochemical and Biophysical Research Communications388(2009)418–421421。
曼氏无针乌贼墨汁黑色素对免疫低下模型小鼠的调节作用
曼氏无针乌贼墨汁黑色素对免疫低下模型小鼠的调节作用董慧;王力东;王春琳;宋微微;母昌考;李荣华【摘要】In this study, the immunomodulatory effects of MSMI ( melanin from Sepiella maindroni ink) on hypoimmune mice induced by Hydrocortisone( HYD) was investigated . ICR mice were selected as animal mode, which were randomly divided into six groups, in-cluding control groups, hypoimmune mice, positive control groups( Levamisole Hydrochloride) , and MSMI treated groups of low, me-dium and high dosage. Except for the control group, all the other groups were treated with Hydrocortisone by subcutaneous injection. After a trial period of 10 days, the liver and spleen indexes and phagocytic capacity of macrophages were analyzed, and which were found to be significantly enhanced in the treated groups(P<0. 01,P<0. 01),(P<0. 05,P<0. 01) as compared to that in the control group. Antioxidant enzyme activities were also measured and they significantly increased compared to the hypoimmune mice group. However, lipid peroxidation assayed as melondialdehyde ( MDA) reduced significantly in the treated groups compared to that in the control. It is suggested that MSMI can promote immune function of immune-depressed mice.%探究曼氏无针乌贼墨汁黑色素( Melanin from Sepiella Maindroni Ink,MSMI)对免疫低下小鼠的调节作用。
纳米材料在生物医学领域的应用英文文献综述
纳米材料在生物医学领域的应用英文文献综述There have been numerous studies on the applications of nanomaterials in the field of biomedical sciences. These materials have unique properties at the nanoscale, which make them attractive for use in various biomedical applications. In this review, we will summarize some of the recent studies on the applications of nanomaterials in the biomedical field.Nanoparticles have been widely investigated for drug delivery purposes. These particles can encapsulate drugs and provide controlled release, improving drug efficacy and reducing side effects. For example, liposomes and polymeric nanoparticles have been used to deliver anticancer drugs specifically to tumor cells, thereby reducing systemic toxicity. Additionally, gold nanoparticles have been utilized to enhance the delivery of therapeutic molecules, such assmall interfering RNA (siRNA), through their ability to penetrate cell membranes.In the field of tissue engineering, nanomaterials have been employed to develop scaffolds with enhanced properties for tissue regeneration. By modifying the surface properties of nanomaterials, researchers have been able to promote cell adhesion and proliferation. For instance, carbon nanotubes and nanofibers have been incorporated into scaffolds to enhance mechanical strength and electrical conductivity, which are essential for certain tissue engineering applications.Furthermore, nanomaterials have been used in the development of diagnostic tools, such as biosensors and imaging agents. Engineered nanoparticles can be functionalized with specific ligands or antibodies to selectively bind to disease markers, enabling the detection and monitoring of various diseases. In addition,superparamagnetic nanoparticles have been employed as contrast agents for magnetic resonance imaging (MRI),offering better visualization of anatomical structures and disease sites.Nanomaterials have also shown promise in the field of theranostics, which involves simultaneous diagnosis and therapy. By incorporating therapeutic agents and imaging agents into a single nanoplatform, researchers have been able to develop theranostic nanoparticles that can target specific cells or tissues, deliver therapy, and monitor treatment response in real-time.In conclusion, nanomaterials have demonstratedsignificant potential in the field of biomedical sciences. Their unique properties make them suitable for applicationsin drug delivery, tissue engineering, diagnostics, and theranostics. Continued research and development in thisfield will likely lead to further advancements and innovative applications of nanomaterials in medicine.。
牛骨髓蛋白的酶解工艺优化及其理化性质和抗氧化特性
古丽米热·阿巴拜克日，帕尔哈提·柔孜，则拉莱·司玛依，等. 牛骨髓蛋白的酶解工艺优化及其理化性质和抗氧化特性[J]. 食品工业科技，2023，44（20）：171−181. doi: 10.13386/j.issn1002-0306.2022100246ABABAIKERI Gulimire, ROZI Parhat, SEMAYI Zelalai, et al. Optimization of Enzymatic Hydrolysis of Bovine Bone Marrow Protein and Its Physicochemical and Antioxidant Properties[J]. Science and Technology of Food Industry, 2023, 44(20): 171−181. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022100246· 工艺技术 ·牛骨髓蛋白的酶解工艺优化及其理化性质和抗氧化特性古丽米热·阿巴拜克日，帕尔哈提·柔孜*，则拉莱·司玛依，阿力木·阿布都艾尼，曹 博，杨晓君（新疆农业大学食品科学与药学学院，新疆乌鲁木齐 830052）摘 要：探讨牛骨髓蛋白(Bovine bone marrow protein, BBMP)酶解工艺并评价其理化性质和抗氧化活性，挖掘其潜在药用和保健功效物质基础，提升牛骨的综合利用价值。
以水解度（DH ）、蛋白含量、1,1-二苯基-2-三硝基苯肼自由基（DPPH·）清除率为评价指标，结合结构表征，筛选最佳酶种。
以酶解时间、酶添加量、pH 、酶解温度为自变量，采用响应面法优化牛骨髓蛋白的酶解工艺，并研究其酶解物的理化性质和抗氧化活性。
药理学英文文献
Stroke is the second leading cause of death in industrialized countries and the most important cause of acquired adult disability.1) The only current effective available stroke therapy is Thrombolysis but it is limited to only about 5% of stroke patients as it carries the risk of intracranial hemorrhagic transformation.2) Therefore, neuroprotective agents with an extended therapeutic window and the ability to prevent multiple neurochemical cascades which ultimately cause irreversible brain damage are urgently required.
Jia QI,a,b Zhen Yi HONG,a Hong XIN,a and Yi Zhun ZHU*,a,c
a Department of Pharmacology, School of Pharmacy and Institute of Biomedical Sciences, Fudan University; Shanghai 201203, China: b Deparment of Pharmacy, Xinhua Hospital Affiliated to Shanghai Jiaotong University; Shanghai 200092, China: and c Department of Pharmacology, Yong Loo Lin School of Medicine, Centre of Life Sciences, National University of Singapore; 28 Medical Drive, 117456, Singapore. Received June 19, 2010; accepted August 28, 2010; published online September 17, 2010
朱章志运用扶正祛邪法论治糖尿病经验
ʌ临证验案ɔ朱章志运用扶正祛邪法论治糖尿病经验❋曾绘域1,朱章志2ә,周㊀海3,陈㊀珺3,张文婧3(1.深圳市中西医结合医院,广东深圳㊀518104;2.广州中医药大学第一附属医院,广州㊀510405;3.广州中医药大学,广州㊀510405)㊀㊀摘要:糖尿病属于中医学 消渴病 范畴,以往医家多认为其病机为阴虚燥热,治疗以滋阴清热为法㊂朱章志教授通过长期的临床观察与实践,立足于张仲景 保胃气,扶阳气 的理论,认为糖尿病的病机为正虚邪滞,即太阴虚损㊁阳气不足㊁收敛不及,寒㊁水㊁湿之邪阻滞阳气运行通道㊂治疗上不囿陈法,以扶正祛邪为大法,通过固护太阴㊁扶助阳气㊁收敛阳气,祛除寒水湿之邪,恢复阳气运行之通畅,使阳气功能复常㊁运行有序,为糖尿病的治疗提供临床新思路㊂㊀㊀关键词:扶正祛邪;糖尿病;朱章志㊀㊀中图分类号:R587.1㊀㊀文献标识码:A㊀㊀文章编号:1006-3250(2021)01-0149-03Discussion on ZHU Zhang-zhi's Experience in Treating Diabetes Mellitus by Using The Method of Reinforcing The Healthy Qi and Eliminating The Pathogenic FactorsZENG Hui-yu 1,ZHU Zhang-zhi 2ә,ZHOU Hai 3,CHEN Jun 3,ZHANG Wen-jing 3(1.Shenzhen Hospital of Integrated traditional Chinese and Western Medicine,Guangdong,Shenzhen 518104,China;2.The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;3.Guangzhou University of Chinese Medicine,Guangzhou 510405,China)㊀㊀Abstract :Diabetes mellitus belongs to the category of "xiao ke"in traditional Chinese medicine.Doctors used to think that its pathogenesis was Yin deficiency and dryness heat ,and the treatment was nourishing Yin and clearing heat.Through long-term clinical observation and practice ,and based on ZHANG Zhong-jing's theory of protecting stomach Qi and supporting Yng Q ,professor ZHU Zhang-zhi believes that the pathogenesis of diabetes is deficiency of healthy Qi and stagnation of pathogen.Because of the deficiency of greater Yin and Yang Qi ,and the lack of convergence ,the cold ,water and dampness block the operational channel of Yang Qi.The treatment of diabetes mellitus should be based on reinforcing the healthy Qi and eliminating the pathogenic factors.By strengthening Taiyin ,supporting Yang Qi ,astringent Yang Qi ,dispelling the evil of cold ,water and dampness ,we can restore the smooth operation of Yang Qi ,restore the function of Yang Qi to normal and operate orderly ,which provides a new clinical method for the treatment of diabetes mellitus.㊀㊀Key words :Reinforcing the healthy Qi and eliminating the pathogenic factors ;Diabetes mellitus ;ZHU Zhang-zhi❋基金项目:国家自然科学基金资助项目(81873190)-降糖三黄片在糖脂毒性所致胰岛β细胞损伤的自噬调控作用作者简介:曾绘域(1990-),女,广东云浮人,住院医师,硕士研究生,从事六经辨治内分泌疾病的临床与研究㊂ә通讯作者:朱章志(1963-),男,湖南衡阳人,主任医师,博士研究生导师,从事六经辨治内分泌疾病的临床与研究,Tel :************,E-mail :zhuangi@ ㊂㊀㊀随着人口老龄化和生活方式的改变,我国糖尿病的患病率呈上升趋势,2013年我国18岁以上人群糖尿病患病率为10.4%[1]㊂中医药在延缓糖尿病的进展及防治其并发症方面具有一定优势[2-4]㊂糖尿病属于中医学 消渴病 范畴,以往医家多认为其病机为阴虚燥热,治疗以滋阴清热为法,但疗效尚不能令人满意㊂朱章志教授通过长期的临床观察与实践,认为正虚邪滞乃糖尿病病机之核心,采用扶正祛邪法治之屡获奇效㊂1㊀正虚邪滞之糖尿病病机‘素问㊃经脉别论篇“曰: 饮入于胃,游溢精气,上输于脾,脾气散精 水精四布,五经并行㊂食物入胃,经脾胃运化化生精气,然后输布全身㊂糖尿病患者常嗜食肥甘,起居无常,烦劳紧张,致太阴虚损,正气内虚,阳气戕伐,津液代谢异常,而生寒水湿之邪㊂寒㊁水㊁湿之邪气作为阴邪,又可阻滞阳气运行之通道㊂阳气运行通道不畅,不能敷布温煦四肢,可见手足逆冷;阳气运行受阻,又可出现郁而化热之象㊂因此朱章志认为,疗糖尿病的关键在于恢复阳气运行之通畅,根据糖尿病正虚邪滞的病机,治疗以扶正祛邪为法,顾护太阴㊁扶助阳气㊁收敛阳气,祛除寒水湿之邪,使阳气功能复常则行有序㊂2㊀运用扶正祛邪法治疗糖尿病2.1㊀扶正2.1.1㊀固护中气,扶助阳气㊀张仲景遣方用药常体现 保胃气 之思想[5],如桂枝汤中配伍生姜㊁大枣㊁炙甘草,发汗祛邪不忘顾护中气;又如白虎汤中加梗米㊁炙甘草以和中益胃,又可防止石膏㊁知母大寒伤中㊂ 有胃气则生,无胃气则死 ,故扶正之要以保胃气为先㊂朱章志认为,阳气在人体的生命活动中占主导9412021年1月第27卷第1期January 2021Vol.27.No.1㊀㊀㊀㊀㊀㊀中国中医基础医学杂志Journal of Basic Chinese Medicine地位㊂‘素问㊃生气通天论篇“曰: 阳气者若天与日,失其所则折寿而不彰 是故阳因而上,卫外者也㊂ ‘黄帝内经“把阳气比作太阳,阳气运行失常可致短寿㊂阳气具有抵御外邪㊁护卫生命㊁维持机体生命活动的作用,津液的气化㊁血液的运行均需阳气的温煦与推动㊂因此,在人体的阴阳平衡中阳气起着主导作用㊂朱章志认为,正气虚衰㊁太阴虚损㊁阳气不足是糖尿病发生发展之根本原因,因此扶正首当 固护中气,扶助阳气 ,故常以附子理中汤为底方,固护中宫㊂太阴脾土居中央,犹如足球比赛之中场,能联系前锋与后卫进可攻退可守,进可充养肺卫之气抵御外邪,退可顾护少阴以防寒邪内陷㊂‘四圣心源㊃卷二太阴湿土“提到: 湿者,太阴土气之所化也故胃家之燥,不敌脾家之湿,病则土燥者少而土湿者多也㊂[6] 阴脾土易挟寒湿,附子理中汤功善固护中气㊁温补脾阳而散寒湿,为治疗太阴阳虚寒湿之要方㊂方中附子辛温大热,补坎中真阳,又能散寒湿,荡去群阴;干姜去脏腑沉寒痼冷,温暖脾土,复兴火种;人参被誉为 百草之王 能大补元气,为扶正固本之极品;白术味苦性温,功善健脾燥湿,乃扶植太阴之要药;炙甘草善益气补中,调和药性,诸药合用以收培补中阳㊁散寒除湿之效㊂若其人神疲懒言,气虚较甚,在附子理中汤的基础上可重用红参㊁北芪以大补元气,健脾益气;若其人四肢不温㊁肢体困重㊁寒湿较重者,可加重附子㊁干姜之量,并加细辛㊁吴茱萸以散久寒;若其人口干口苦㊁舌苔黄腻㊁大便黏滞不爽兼夹湿热之象,可仿当归拈痛汤之意,加茵陈㊁当归㊁黄芩以利湿清热㊂2.1.2㊀收敛阳气,阳密乃固㊀朱章志认为, 阴 可理解为 阳气 的收敛㊁收藏状态,糖尿病 阴虚燥热 之象乃阳气不足㊁收敛不及㊁升发太过所致[7]㊂‘素问㊃生气通天论篇“提到: 阳气者,烦劳则张 ㊂现代人起居无节,以妄为常,阳气因而不能潜藏,常常浮越于外容易出现假热之象,医者不察,妄投清热泻火之品,实乃雪上加霜㊂ 凡阴阳之要,阳密乃固 ,收敛阳气即是扶正,犹如太极之能收能放,收敛是为了聚集能量,阳气固密,正气才能强盛,方能更好的制敌㊂朱章志常用砂仁㊁肉桂㊁白芍㊁山萸肉㊁泽泻等药物收敛阳气㊂砂仁辛温,既能宣太阴之寒湿,又能纳气归肾,使阳气收敛于少阴,少火生气㊂‘本草经疏“提到: 缩砂蜜,辛能散,又能润 辛以润肾,故使气下行 气下则气得归元㊂[8] 肉桂引火归原,导浮越之阳气归于命门,益火消阴㊂若患者出现咽痛㊁牙龈肿痛㊁痤疮等阳气不敛㊁虚火上冲之象,常用砂仁㊁肉桂以收敛阳气,纳气归肾,引火归原㊂白芍味酸能敛,敛降甲木胆火,使相火归位㊂‘本草求真“曰: 气之盛者,必赖酸为之收,故白芍号为敛肝之液,收肝之气,而令气不妄行也㊂[9] 朱章志常使用白芍以补肝之体㊁助肝之用,收敛肝气,肝平则郁气自除,火热自消㊂山萸肉秘精气㊁敛阳气,使龙雷之火归于水中㊂朱章志常用山萸肉收敛正气,遇汗出多者,常重用以固涩敛汗㊂泽泻能泻能降,能入肾泻浊,开气化之源,泻浊以利扶正,又能降气而引火下行㊂朱章志常用泽泻打通西方潜藏之要塞[10],在温阳之品中加入泽泻,利于阳气潜藏,使孤阳有归㊂2.1.3㊀填补阴精,以滋化源㊀‘素问㊃金匮真言论篇“提到: 夫精者,身之本也㊂ 精 是人体生命活动的物质基础,能化气生髓,濡养脏腑㊂人体之精禀受于父母,又由后天水谷之精不断充养,归藏于肾中㊂ 孤阴不生,独阳不长 ,无阳则阴无以生,无阴则阳无以化㊂肾乃水火之脏,阴精充足才能涵养坎中真火,使真阳固密于内,化生正气㊂朱章志常在秋冬之季嘱糖尿病患者进补阿胶等血肉有情之品填补肾精㊂肾主封藏,秋冬进补使肾精充养,以滋阳气化生之源㊂阿胶用黄酒烊化,既能祛除阿胶之腥,又能借黄酒通行之性解阿胶滋腻碍胃之弊,每日少量服用,以有形之精难以速生,填补肾精以缓补为要㊂除此之外,遣方用药时亦会注意顾护阴精,在使用温阳药的同时常常配伍山萸肉㊁白芍等养阴药,以防温燥伤阴之弊㊂2.2㊀祛邪2.2.1㊀外散寒水以运太阳㊀ 太阳为开 ,太阳乃三阳之表,巨阳也,其性开泄以应天,为祛邪之重要通道㊂在运气里,太阳在天为寒,在地为水,合而为太阳寒水㊂张仲景太阳病篇研究的是水循环过程,治太阳就是治水[11]㊂寒㊁水之邪闭郁在表,气血运行不畅,可见肌肤麻木不仁㊂邪气滞留太阳,阻碍阳气运行,当因势利导㊁开太阳之表以发汗,外散寒㊁水之邪㊂糖尿病患者正气亏虚为本,祛邪不能伤正,朱章志临床常用桂枝麻黄各半汤小发其汗,使玄府开张,邪有出路㊂桂枝麻黄各半汤乃发汗轻剂,为桂枝汤与麻黄汤相合而得,其中麻黄㊁桂枝㊁生姜㊁北杏发散宣肺以开皮毛,芍药㊁大枣㊁炙甘草酸甘化阴以益营,诸药相合,刚柔相济,祛邪而不伤正㊂邪去正安,阳气运行通畅,水液代谢复常则阳气自充,而无寒水之扰㊂若寒邪较重可用三拗汤,此为麻黄汤去桂枝而成,功善开宣肺气,疏散风寒,因去辛温之桂枝发汗力不及麻黄汤,祛邪而不伤正㊂2.2.2㊀下利水湿以健少阴㊀少阴乃水火交会之脏,元气之根,人身立命之本㊂‘医理真传“提到: 坎中真阳,一名龙雷火,发而为病,一名元阳外越,一名孤阳上浮,一名虚火上冲㊂此际之龙,乃初生之龙,不能飞腾而兴云布雨,惟潜于渊中,以水为家,以水为性,遂安其在下之位㊂水盛一分龙亦盛一分,水高一尺龙亦高一尺,是龙之因水盛而游 [12]㊂阴盛051中国中医基础医学杂志Journal of Basic Chinese Medicine㊀㊀㊀㊀㊀㊀2021年1月第27卷第1期January2021Vol.27.No.1则阳衰,水湿之邪泛滥,则龙雷之火因而飞越在外㊂叶天士深谙张仲景之理,提到 通阳不在温,而在利小便 [10,13],通过利小便的方法,使水湿之邪从下而解,阳气运行通道无水湿之邪阻碍,则阳气无需温养而自通,水盛得除则真龙亦安其位㊂朱章志常用五苓散㊁真武汤下利水湿,以复阳气之通达,少阴之健运㊂五苓散具有通阳化气利水之效,治疗膀胱气化不利形成的蓄水证㊂方中猪苓㊁茯苓㊁泽泻导水湿之邪下行;白术健脾燥湿,杜绝生湿之源;桂枝助膀胱气化,通阳化气行水又通气于表,使全身在表之湿邪皆得解,五药合用,膀胱气化复常,水道通调使小便得利,水湿得出㊂真武汤为治疗少阴阳虚㊁水气泛滥之主方,方中附子振奋少阴阳气,使水有所主;白术㊁茯苓健脾制水;生姜助附子敷布阳气,宣散水气;芍药利小便,制附㊁姜之燥,五味相合共奏温阳利水之功㊂2.2.3㊀开郁逐寒以畅厥阴㊀肝为将军之官,肝气主动主升发,能统帅兵马,捍卫君主㊂厥阴肝经,体阴用阳,内寄相火,相火敷布阳气,祛阴除寒,是祛邪的先锋主力军㊂朱章志常用吴茱萸汤祛除厥阴肝经之寒邪,恢复肝经阳气之运行㊂方中吴茱萸辛苦而温,芳香而燥,‘本草汇言“曰: 开郁化滞,逐冷降气之药 [14],能温胃暖肝,降浊阴止呕逆,为治疗肝寒之要药㊂配以生姜温胃散寒,佐以人参㊁大枣健脾益气补虚,全方散寒与降逆并施,共奏暖肝温胃㊁降逆止呕之效㊂‘素问㊃至真要大论篇“说: 帝曰:厥阴何也?岐伯曰:两阴交尽也㊂ 物极必反,重阴必阳㊂厥阴为阴尽阳生之脏,足厥阴肝经与足少阳胆经互为表里,若出现肝气不疏㊁枢机不利㊁气郁化火,朱章志常用小柴胡汤和畅枢机,开郁以复气机调达㊂方中柴胡疏泄肝胆之气;黄芩清泄胆火,一疏一清,气郁通达,火郁得发;生姜㊁半夏和胃降逆;人参㊁大枣㊁炙甘草固护中宫,全方寒温并用㊁补泻兼施,以复厥阴疏泄之职,使气机和畅㊁阳气运行有序㊂3㊀典型病案患者杨某,女,65岁,2017年3月10日初诊:2型糖尿病病史6年余,症见疲乏,双下肢轻度浮肿,下肢冰凉,背部易汗出,口苦口干,偶有腰膝酸软,纳眠可,二便调,舌淡暗,苔黄腻,脉沉细㊂辅助检查示糖化血红蛋白10.8%,空腹血糖14.59mmol/L,总胆固醇6.38mmol/L,甘油三酯3.66mmol/L,低密度脂蛋白胆固醇4.34mmol/L㊂西医诊断2型糖尿病㊁高脂血症,治疗给予门冬胰岛素30(早餐前22u晚餐前20u)控制血糖,阿托伐他汀钙片(20mg, qn)调脂㊂中医诊断消渴病,少阴阳虚寒湿证㊂患者少阴阳气衰微不足以养神,固见疲乏;腰为肾之府,少阴阳虚则见腰膝酸软,阳虚寒盛则见下肢冰凉;背部正中乃督脉运行之所,阳气虚衰无以固摄则见背部汗出;少阴阳虚不能主水,寒水泛滥则见双下肢浮肿;水湿内停有郁而化热之象,则见口苦口干㊁舌苔黄腻㊁舌淡暗,脉沉细为少阴阳虚寒湿之征,治以温阳散寒㊁利水除湿为法㊂方以扶正祛邪方合当归拈痛汤加减:炮附片10g(先煎1h),红参10g (另炖),干姜15g,白术30g,炙甘草15g,桂枝12 g,麻黄8g,生姜30g,猪苓10g,泽泻30g,茯苓30 g,白芍30g,酒萸肉45g,当归15g,茵陈10g,5剂水煎服,2d1剂,水煎至250ml,饭后分2次服用,次日复煎㊂方中以附子理中汤为底方温补中焦,散寒除湿;加桂枝㊁麻黄使寒湿之邪从皮毛而解;加五苓散通阳化气,使湿邪从下而出;生姜散寒除湿;白芍㊁酒萸肉收敛阳气,以助正气祛邪;当归活血利水;茵陈清热利湿㊂2017年3月24日二诊:患者双下肢浮肿减轻,疲乏较前好转,无口干口苦,无腰膝酸软,仍觉下肢冰凉,背部仍有汗出,动则尤甚,大便每日二行,质偏烂,舌淡暗,苔白腻,脉细㊂患者大便质烂,乃邪有出路,导水湿之邪从大便而解㊂患者无口干口苦,舌苔由黄腻转为白腻,知湿郁化热之象已除,遂去茵陈㊂仍觉下肢冰凉乃内有久寒,加制吴茱萸12g以散沉寒痼冷;上方加酒萸肉至60g以加强收敛阳气㊁固摄敛汗之效,加黄芪60g以健脾益气敛汗;加砂仁6g(后下)㊁肉桂3g(焗服)以加强收敛阳气㊁扶助正气之效,7剂水煎服,服法同前㊂2017年4月7日三诊:患者背部汗出减少,下肢转温,余症皆除,大便每日二行质软,舌淡红,苔薄白,脉细较前有力,继续服二诊方药5剂㊂后给予附子理中丸(每次8粒,每日3次)服用1个月巩固疗效㊂2017年11月17日复诊:患者上述症状皆除,纳眠可,二便调㊂复查糖化血红蛋白6.8%,空腹血糖6.5mmol/L,总胆固醇5.14mmol/L,甘油三酯1.65 mmol/L,低密度脂蛋白胆固醇2.43mmol/L㊂4㊀结语以往医家多以滋阴清热为法治疗糖尿病,通过长期的临床实践,朱章志不囿陈法,根据糖尿病患者当前之病因病机特点,运用扶正祛邪法治疗糖尿病,通过顾护太阴㊁扶助阳气㊁收敛阳气,祛除寒水湿之邪气,恢复阳气运行之通畅,为糖尿病的治疗提供新思路㊂参考文献:[1]㊀WANG L GAO-P-ZHANG-M,et al.Prevalence and EthnicPattern of Diabetes and Prediabetes in China in2013[J].JAMA,2017,317(24):2515-2523.[2]㊀谭宏韬,刘树林,朱章志,等. 首辨阴阳,再辨六经 论治惠州地区2型糖尿病的临床观察[J].中华中医药杂志,2018,33(9):4240-4244.(下转第181页)offspring of sleep-deprived mice[J].Psychoneuroendocrinology,2009,35(5):775-784.[9]㊀覃甘梅,覃骊兰.心肾不交型失眠动物模型研究进展[J].中华中医药杂志,2018,33(1):229-231.[10]㊀吕志平,刘承才.肝郁致瘀机理探讨[J].中医杂志,2000,41(6):367-368.[11]㊀游秋云,王平,田代志,等.老年肝郁失眠证候大鼠模型的建立和评价[J].中国实验方剂学杂志,2013,19(2):222-225. [12]㊀唐仕欢,杨洪军,黄璐琦. 以方测证 方法应用的反思[J].中国中西医结合杂志,2007,27(3):259-262.[13]㊀卢岩,刘振华,于晓华,等.疏肝调神针法针刺对睡眠剥夺模型大鼠神经递质的影响[J].山东中医杂志,2017,36(4):322-325. [14]㊀YANG CR,SEAMANS JK,GORELOVA N.Developing aneuronal model for the pathophysiology of schizophrenia based onthe nature of electrophysiological actions of dopamine in theprefrontal cortex[J].Neuropsychopharmacology,1999,21(2):161-194.[15]㊀何林熹,诸毅晖,杨翠花,等.失眠肝郁化火证大鼠模型的建立及其评价[J].中华中医药杂志,2018,33(9):3890-3894. [16]㊀李越峰,徐富菊,张泽国,等.四逆散对大鼠睡眠时相影响的实验研究[J].中国临床药理学杂志,2014,30(10):936-938. [17]㊀张晓婷,刘文超,刘俊昌,等.电击法建立SD大鼠焦虑型心理应激-失眠模型的研究[J].现代中西医结合杂志,2018,27(30):3316-3319.[18]㊀钱伯初,史红,郑晓亮.新的失眠动物模型研究概述[J].中国新药杂志,2008,17(1):1-4.[19]㊀朱洁,申国明,汪远金,等.肝郁证失眠大鼠模型的建立与评价[J].中医杂志,2011,52(8):689-692.[20]㊀刘倩,李蜀平,廖磊,等.调和肝脾方治疗失眠的实验研究[J].北京中医药,2018,37(8):768-770.[21]㊀全世建,焦蒙蒙,黑赏艳,等.交泰丸对睡眠剥夺大鼠下丘脑Orexin A及γ-氨基丁酸的影响[J].广州中医药大学学报,2015,32(1):103-105.[22]㊀KOBAN M,SWINSON KL.Chronic REM-sleep deprivation ofrats elevates metabolic rate and increases UCP1gene expressionin brown adipose tissue[J].Am J Physiol Endocrinol Metab,2005,289(1):68-74.[23]㊀赵俊云,杨晓敏,胡秀华,等.失眠动物模型HPA轴和表观遗传修饰的变化及交泰丸的干预作用[J].中医药学报,2018,46(4):19-21.[24]㊀GORGULU Y,CALIYURT O.Rapid antidepressant effects ofsleep deprivation therapy correlates with serum BDNF changes inmajor depression[J].Brain Res Bull,2009,80(3):158-162.[25]㊀BENCA RM,PETERSON MJ.Insomnia and depression[J].Sleep Med,2008,9(1):S3-S9.[26]㊀郜红利,涂星,卢映,等.心肾不交所致失眠大鼠模型[J].中成药,2014,36(6):1138-1141.[27]㊀杨钰涵,孙雨,王珺,等.中医病证相符的大鼠心肾不交失眠模型的建立及其血清代谢组学研究[J].中国中药杂志,2020,45(2):383-390.[28]㊀石皓月,鲁艺,李钰昕,等.中药治疗对氯苯丙氨酸失眠模型大鼠影响的基础研究进展[J].中国医药导报,2018,15(11):33-36.[29]㊀全世建,何树茂,钱莉莉.交泰丸交通心肾治疗失眠作用机理研究[J].辽宁中医药大学学报,2011,13(8):12-14. [30]㊀GULEC M,OZKOL H,SELVI Y,et al.Oxidative stress inpatients with primary insomnia[J].Pro NeuropsychopharmacolBiol Psychiatry,2012,37(2):247-251.[31]㊀ZHANG H,CAO D,CUI W,et al.Molecular bases ofthioredoxin and thioredoxin reductase-mediated prooxidant actionsof(-)-epigallocatechin-3-gallate[J].Free Radic Biol Med,2010,49(12):2010-2018.[32]㊀谢光璟,刘源才,胡辉,等.基于Trx系统介导的抗氧化应激探讨天王补心方对失眠模型大鼠的干预作用[J].时珍国医国药,2019,30(4):805-808.[33]㊀黄攀攀,王平,李贵海,等.老年阴虚失眠动物模型的建立与评价[J].中华中医药学刊,2010,28(8):1719-1723.[34]㊀XIONG L,HUANG XJ,SONG PX.The experiment ofstudymodel of Deficiency of yin Insomnia by Yangyin anshenkoufuye[J].Chin J Pract Chin Mod Med,2005,18(18):1187-1188.[35]㊀韦祎,唐汉庆,李克明,等.脾阳虚证失眠大鼠模型的建立和附子理中汤的干预效应[J].中国实验方剂学杂志,2013,19(16):289-292.[36]㊀王志鹏.桂枝甘草龙骨牡蛎汤对阳虚证失眠大鼠脑内5-HT㊁NE含量的影响[D].南京:东南大学,2015.[37]㊀MURRAY NM,BUCHANAN GF,RICHERSON GB.InsomniaCaused by Serotonin Depletion is Due to Hypothermia[J].Sleep,2015,38(12):1985-1993.[38]㊀宋亚刚,李艳,崔琳琳,等.中医药病证结合动物模型的现代应用研究及思考[J].中草药,2019,50(16):3971-3978. [39]㊀李晓娟,白晓晖,陈家旭,等.中医动物模型研制方法及展望[J].中华中医药杂志,2014,29(7):2263-2266.[40]㊀刘臻,谢晨,赵娜,等.失眠动物模型的制作与评价[J].中医学报,2013,28(12):1846-1848.收稿日期:2020-05-18(上接第151页)[3]㊀司芹芹,牛晓红,杨海卿,等.温阳益气养阴活血方治疗2型糖尿病肾病的临床疗效[J].中华中医药学刊,2018,36(3):703-705.[4]㊀郭仪,石岩.中药复方治疗糖尿病大血管病变临床疗效及对血糖㊁血脂影响的系统评价[J].中华中医药学刊,2017,35(6):1369-1375.[5]㊀方春平,刘步平,朱章志.‘伤寒论“中 胃气 思想在病脉辨证中的运用[J].浙江中医药大学学报,2014,38(8):948-950.[6]㊀黄元御.四圣心源[M].北京:中国中医药出版社,2009:24.[7]㊀朱章志,林明欣,樊毓运.立足 阳主阴从 浅析糖尿病的中医治疗[J].江苏中医药,2011,43(4):7-8.[8]㊀缪希雍.本草经疏[M].北京:中国医药科技出版社,2011:56.[9]㊀黄宫绣.本草求真[M].北京:中国中医药出版社,2008:132.[10]㊀林明欣,裴倩,朱章志.浅析 通阳不在温,而在利小便 [J].中医杂志,2011,52(19):1705-1706.[11]㊀刘力红.思考中医[M].桂林:广西师范大学出版社,2006:457.[12]㊀郑寿全.医理真传[M].北京:中国中医药出版社,2008:3.[13]㊀刘涛,张毅,李娟,等.结合‘伤寒论“探讨 通阳不在温而在利小便 [J].中国中医药信息杂志,2017,24(9):106-107.[14]㊀倪朱谟.本草汇言[M].北京:中医古籍出版社,2010:87.收稿日期:2020-04-27。
氯法齐明治疗耐多药结核病有效性与不良反应的研究进展
氯法齐明治疗耐多药结核病有效性与不良反应的研究进展桂敏陈敬芳邓国防付亮曾谷清【摘要】氯法齐明是治疗耐多药结核病领域中老药新用的代表性药品，可有效增加耐多药结核病的痰菌阴转 率、病灶吸收率及空洞缩小率，且不增加药物不良反应发生率，是世界卫生组织推荐治疗耐多药结核病的重要药 品作者对氯法齐明治疗耐多药结核病的临床有效性和不良反应进行简要综述.旨在为临床用药和临床研究提供参考。
【关键词】结核，抗多种药物性；氯法齐明；药物毒性Research progress on the effectiveness and adverse reactions of clofazimine in the treatment of multidrug-resistant tuIxTculosis GUI M i n', CHEN J in g-fm ig1'2 ■,DENG G uo-fang1. FU L ia n g1, ZENG G u-qitig1. 1School o f N ursing, University o f South China , H unan Province, H mgyan^ 421001. China-, ' C rude/ff Hospitals Create O ffice, Nfiliiimil Clinical Resemrh Center for Infectious Disease, the Third Pm lilp's Husliilal o f Shenzhen , Shenzhen 5hS112, China-, 1the Sm m d Department of Pulmonary Diseases, the Third People' s Hospital o f Shenzhen, Shenzhen ^1SI I2,ChinaCm'mpmiding uuttwr:CHEN Jing-j'm g, Em nil:138****9640@163.cum【Abstract】Clofazimine is a representative repurposed drug in the field of multidrug-resistant tuberculosis (MDR-TB). Clofazimine can effectively increase the sputum negative conversion, lung lesion absorption, and cavity reduction of MDR-TB, without increasing the incidence of adverse reactions. It is an important drug recommended by the WHO guidelines. We briefly reviewed the clinical application progress and adverse reaction management of clofazimine in the treatment of MDR-TB. in order to provide references for clinical medication and clinical research.【Key words】Tuberculosis，multidrug-resistant; Clofazimine; Drug toxicity耐药结核病疗程长、治愈率低.已成为重大公共卫生问 题•研发抗结核新药和探索治疗新方案迫在眉睫。
碱性糊精酶生产菌Bacillus flexus PR1的筛选及其初步表征说明书
Int.J.Curr.Microbiol.App.Sci (2017) 6(5): 2981-2987Original Research Article https:///10.20546/ijcmas.2017.605.338 Isolation and Partial Characterization of Amylase ProducingBacillus flexus PR1P. M. Patel1, R. G. Chaudhari2* and S. A. Bhatt31M.G. Science Institute, Ahmedabad, Gujarat, India2Shri PJPN Science College, Maktupur, Unjha, Gujarat, India3Department of Life Sciences, HNGU, Patan, India*Corresponding authorA B S T R A C TIntroductionBacteria of the genus Bacillus are potentially rich source of various valuable enzymes. These bacteria can produce extracellular enzymes with major applications in processes operated under extreme conditions due to their adaptability to environmental limits Alkaliphilic Bacillus species are considered to be of major interest due to their ability to produce extracellular enzymes that are active and stable at high pH, including alkaline amylase, protease and cellulase. The unusual properties of these enzymes propose a potential opportunity for their utilization in processes demanding such extreme conditions (Karnwal and Nigam, 2013). Amylases (E.C.3.2.1.1) are enzymes which hydrolyze starch molecules to give diverse products including dextrins and even smaller polymers composed of glucose units.Amylases constitute a class of industrial enzymes, which contribute approximately 25% of the enzyme market covering many industrial processes such as sugar, textile, paper, brewing, distilling industries and pharmaceuticals (Das et al., 2011). Thus, the potency of alkaline amylases for industrial applications have encouraged the search for microbial strains expressing activities with enzyme with desired properties (Alkando etInternational Journal of Current Microbiology and Applied SciencesISSN: 2319-7706 Volume 6 Number 5 (2017) pp. 2981-2987Journal homepage: Bacillus strains were isolated from highly alkaline soil from Himatnagar city of North Gujarat. The best amylase producing bacteria were screened on starch agar plate. Among the screened isolates highest amylase producer strain PR-I was selected, characterized and identified on the basis of 16S r -RNA sequence homology. The major cellular fatty acids of the strain were also identified. The strain was gram positive, spore forming and aerobic, motile bacillus species. 16S r- RNA sequence homology shows 99% similarity with Bacillus flexus. This strain grows in the pH range of 4.0 to 12. The optimum growth was obtained at pH 10 and temperature 370 C. PR-1 produced extracellular alkaline amylase, and its maximal enzyme activity was observed at 450C and10.0 pH.70% enzyme activity was retained at pH 12. The strain showed salt tolerance up to 4% of NaCl. The enzyme activity was found to be increased in the presence of Co2+ and Mg2+ cations. The results of the growth and enzyme activity were discussed.al.,2011). The strain in study PR-1 was originally isolated from alkaline soil in the Himatnagar city, it showed very high alkali-tolerance and was identified as a new member of Bacillus flexus. This work deals with the isolation and characterization of PR-1 and some properties of its extracellular alkaline amylase. The above features could favour current industrial demand of commercially utilizable saccharifying enzymes.Materials and MethodsIsolation and screening of the amylase producing bacteriaTwenty Bacillus spp. capable of growing highly alkaline conditions were isolated from alkaline soil (pH 11.0) in Himatnagar city. Screening for starch hydrolysis activity among the isolated colonies was performed by plating them on the agar medium containing (W/V%): soluble starch 1.0, peptone 1.0, yeast extract 0.5, K2HPO4 0.1, MgSO4·7H2O 0.02, NaCl 1.0, agar 2, pH 10.0 and incubating for 24 h at 37°C. The plates were flooded with iodine reagent to reveal the zone of starch hydrolysis (Van der Maarel et al., 2002).The diameter of the halo zone against the diameter of the colony was used as a semi quantitative method for the selection of the starch hydrolyzing strains. Depending on the zone diameter and clearance isolate PR-I was selected as a good alkaline amylase producer.Phenotypic and genotypic identificationIdentification of selected isolate was carried out on the basis of their morphological and biochemical tests were performed according to Bergey’s manual of systematic bacteriology and the methods described in the Genus Bacillus (Das et al., 2004). It was then confirmed with 16S r RNA sequencing, pure cultures of the target bacteria were grown in nutrient broth medium on a rotary shaker (150 rpm) at 30°C for 24 h for the isolation of genomic DNA (Yadav et al., 2009).Characteristics of amylaseEnzyme production mediumThe medium for enzyme production used having (W/V%): starch 2.0, peptone 1.0, yeast extract 1.0, K2HPO40.1, and MgSO4·7H2O 0.02 (Kaur et al., 2012). The medium (50 mL) was inoculated with 0.5 mL of the inoculum with an optical density of 0.6 at 600 nm and incubated at 37°C (El-Tayeb et al., 2007). Samples were harvested till 72 h by centrifugation at 6000×g for 10 min and the cell free supernatant was analysed for amylase activity as described below. Enzyme assayThe amylase activity was estimated on the basis of the reduction in blue color intensity resulting from enzymatic hydrolysis of starch. The reaction mixture containing 1% starch (0.1 mL), 0.3 mL of 0.05 mol/L glycine NaOH buffer (pH 10.0) and 0.1 mL of culture filtrate was incubated at 37°C for 15 min in a water bath. The reaction was terminated by adding 1 mL chilled 1 mol/L HCl. To this mixture, 2 mL of diluted iodine was added and the volume was made to 10 mL with double distilled water.A substrate control-lacking enzyme was also kept with each set of reaction. The amylase activity was estimated after appropriate dilution and absorbance was read at 620 nm against a substrate blank. One dextrinizing unit of amylase activity is defined as the amount of enzyme that results in 10% decline in the optical density of the starch iodine complex at 620 nm when compared to substrate blank. All the experiments were carried out in triplicates and results presented are the mean of three values.Partial purificationThe cell free supernatant fluid was precipitated using ammonium sulphate to85% saturation. The precipitate was dissolved in 0.2 M glycine NaOH buffer (pH 10.0) and dialysed overnight against the same buffer. The dialysate was used for all enzyme characterization studies.Effect of pH on enzyme activity and stabilityThe pH optimum of the enzyme was determined by varying the pH of the assay reaction mixture using the following buffers (0.2 mol/L): Na2HPO4/citrate phosphate buffer (pH 3.0−8.0), glycine/NaOH buffer (pH 9.0−10.0), Na2HPO4/NaOH buffer (pH 11.0) and KCl/NaOH buffer (pH12.0−13.0). To determine the stability of amylase, the enzyme was pre-incubated in different buffers (pH 7.0−13.0) for 30 and 60 min. The residual enzyme activity was determined as described earlier.Effect of temperature on enzyme activity and stabilityThe temperature optimum of the enzyme was estimated by measuring the amylase activity at different temperatures (40°C −80°C) in 0.2 mol/L glycine/NaOH buffer (pH 10.0).The effect of temperature on amylase stability was determined by measuring the residual activity after 20, 40 and 60 min of preincubation in 0.2 mol/L glycine/NaOH buffer (pH10.0), at temperatures ranging from 40°C to 70°C (Vaseekaran et al., 2010) Effect of metal ions on enzyme activityFor determining the effect of metal ions on amylase activity, enzyme assay was performed after pre-incubation, at 40°C (optimum) for 60 min, of the enzyme with various metal ions each at a concentration of 50 mmol/L. The enzyme assay was carried out in the presence of CaCl2·2H2O, MgSO4·7H2O, FeSO4, CoCl2, MnSO4·4H2O, ZnSO4·7H2O, CuSO4 and EDTA.Results and DiscussionMorphological, physiological and biochemical propertiesTwenty Bacillus spp. capable of growing highly alkaline conditions were isolated from alkaline soil (pH 11.0) in Himatnagar city, among these we have screened a potent alkaline amylase-degrading microorganism (Alkando et al., 2011).Depending on the zone diameter and clearance, the isolate PR-I was selected as a good alkaline amylase producer. The isolate was able to grow and produce extracellular amylase when it was cultured in amylase induction medium (Kanimozhi et al., 2014). It was a Gram-positive, aerobic, motile, spore-forming, alkaliphilic bacterium, and its cells were rod-shaped. It grew at a wide range of pH values (4.0−12) in LB medium, with the optimum being at pH 10. Growth occurred at 10°C−60°C, with the optimum growth temperature being at 37°C. Its NaCl tolerance was high up to 4 %. It showed catalase-positive and oxidase negative reactions but did not reduce nitrate to nitrite Table 1.16S rDNA sequences analysis and (G+C) content were done and to understand the phylogenetic position of our isolate, we constructed a phylogenetic tree based on comparison of 16S rDNA sequences of our isolate and correlative taxa (Fig. 2). PR-I has 99% sequence similarity with all known strains of B. flexus and formed a tight cluster with them. The (G+C) content of the genomic DNA of strain PR-I (39.1mol %) was almostidentical to that of B. flexus (37−39 mol %). Characteristics of amylasepH optima and stability check : Optimum pH of PR-I amylase is shown in (Fig. 1-A). Determination of enzyme activity at 40°C at pH values ranging from 3.0 to 13.0 showed the amylase found to be active in a wide range of pH values with maximum production at pH 10.0 (14.63 U/mL). More than 70% of enzyme activity was found between pH 9.0 and 13.0. A significant decline of enzyme activity was observed in the acidic pH and only 50% residual activity was retained at pH7.0, whereas 90% retained at pH 11.0. The pH stability was determined at 40°C with different pH values (pH 7.0−13.0) (Kamm and Kamm,2004).Fig.1 The effects of temperature and pH on the amylase activity and stabilityFig.1A The effects of pH on the amylaseactivityFig.1C The effects of temperature on theamylase activityFig.1B pH stability of the enzyme Fig.1D Thermal stability of the enzymeTable.1 Physiological and biochemical properties of B flexus PR-ITable.2 Effects of metal ionsAmylase was stable between pH8 and 13, and more than 60% of the activity was retained after incubated for 1h (Fig. 1-B). It is clearly evident that amylase produced by PR-I, which was designated as PR-I amylase, is very stable to extremely alkaline environment. Temperature optimum and stability: Using starch as substrate, the optimum temperature of the amylase was 40°C (Fig.1-C). Under some constant temperature, we found that PR-I amylase was very stable at 50°C (Thippeswamy et al., 2006). Even more than 60% of amylase activity still observed when it was incubated at 60°C for 1 h (Fig.1-D), (Das et al., 2004).Effect of different metal ions and chelating agents: The effects of various metal ions and chelating agents on enzyme activity were examined at pH 10 and 40°C (Table 3). Slight effects were observed in the presence of Zn2+, Fe2+, Cu2+, Mn2+, Ca2+ and chelating agents of EDTA only inhibited activity of the enzyme to 52, 78, 39, 54 and 65%, respectively. In contrast, Mg2+, Co2+obviously increased the enzyme activity to 138 and 123%, respectively (Jogezai et al., 2011) (Table 2).Bacillus species are the most important industrial microorganism with versatile roles. The development of a variety of commercial enzymes products with the desired temperature, pH activity, and stability properties require to be of significant. In this study the isolate PR-I could grow and produce extracellular amylase when it was cultured in amylase agar medium. Combined with morphological, physiological and biochemical characteristics with 16S rDNA sequence analysis and the (G+C) content of the genomic DNA, the strain identified as B.flexus. The 16S rDNA sequences, the hydrolysis of urea, the utilization of D-Xylose, D-Lactose and D- Raffinose, and the excellent pH tolerance, which were different from those of the reference strain B. flexus DSM1320T and the description of the Manuals of Bergey, the strain was classified as a new member of B. flexus. The alkaliphiles can grow at pH 9 and the optimum pH is 10.0−12.0, but they can grow weakly or not grow at pH6.5 approximately. The alkaliphiles can grow at pH 9 and the optimum pH is 10.0−12.0, but they can grow weakly or not grow at pH6.5 approximately (Gangadharan et al., 2006), while alkali- tolerant bacteria cannot grow at the pH over 10.5, but can grow at pH7.0−9.0. Our results indicate that B. flexus can be a potential organism for various applications including the treatment of alkaline agroindustrial wastes.ReferencesAlkando, A. A., Ibrahim, H. M., (2011) A potential new isolate for theproduction of a thermostableextracellular a-amylase. J.Bacteriol.Res. 3, 129–137.Das K, Doley R, Mukherjee A K, (2004) Purification and biochemicalcharacterization of thermostable,alkaliphilic, extracellular α-amylasefrom Bacillus subtilis DM-03, a strainisolated from the traditional food ofIndia. Biotechnol Appl Biochem, 40:291-298.Das S, Singh S, Sharma V, Soni M L (2011) Biotechnological applications ofindustrially important amylaseenzyme. Inter J Phar Bio Sci,2(1):486-496.El-Tayeb O, Mohammed F, Hashem A, Abdullah M (2007) Optimization ofthe industrial production of bacterialalpha amyalse in Egypt. IV. Fermentorproduction and characterization of theenzyme of two strains of Bacillussubtilis and Bacillusamyloliquefaciens. Afr J Biotechnol,7(24):4521-4536Gangadharan D, Sivaramakrishnan S, Namboothiri K M, Pandey A, (2006)Solid culturing of Bacillusamyloliquefaciens for -amylaseproduction. Food Technol Biotechnol,44(2):269-274Jogezai, N., Raza, A., Abbas, F., Bajwa, M., Mohammad, D., Kakar, W., Saeed,M., Awan, A. (2011). Optimization ofcultural conditions for microbial alphaamylase production. J. Microbiol.Antimicrob.3, 221–227.Kamm, B., Kamm, M. (2004). Biorefinery –systems. Chem. Biochem.Eng. 18, 1–6.Kanimozhi, M., Johny, M., Gayathri, N., Subashkumar, R., (2014).Optimization and production of a -amylase from halophilic bacillusspecies isolated from mangrove soilsources. J. Appl.Environ. Microb. 3,70–73.Karnwal, A. and Nigam, V., (2013).Production of amylase enzyme byisolated microorganisms and itsapplication. IJPBS 3, 354–360.Kaur P, Vyas A (2012) Characterization and optimal production of alkaline α-amylase from Bacillus sp. DLB 9. AfrJ Microbiol Res, 6(11):2674-2681 Kumarai, B. L., SaiRam, C. V. S., Kumar, T.S., Sudhakar, P., Vijetha, P., (2011).Screening and isolation ofthermostable a-amylase producingbacteria and optimization of physico-chemical parameters for increasing theyield. Int. J. Pharm. Technol. 3, 1570–1583.Thippeswamy, S., Girigowda, K., Mulimami,H. V., (2006). Isolation andidentification of a- amylase producingBacillus sp. from dhal industry waste.Ind. J. Biochem. Biophys. 43, 295–298.Van der Maarel, M. J. E. C., van der Veen, B.,Uitdehaag, J. C. M., Leemhuis, H.,Dijkhuizen, L., (2002). Properties andapplications of starch converting ofalpha amylase family. J. Biotechnol.94, 137–155.Vaseekaran, S., Balakumar, S., Arasaratnam, V., (2010). Isolation and identificationof a bacterial strain producingthermostable aamylase.Trop. Agric.Res. 22, 1–11.Yadav, V., Prakash, S., Srivastava, S., Verma, P. C., Gupta, V., Basu, V., Rawat, A.K., (2009). Identification ofComamonas species using16S rRNAgene sequence. J. Bioinform. 3, 381–383.。
咖啡因在帕金森病运动并发症的应用
咖啡因在帕金森病运动并发症的应用发表时间：2016-05-23T11:04:08.067Z 来源：《健康世界》2015年12期作者：缪茂军（综述）1，2 陈小武1 陈志斌（审校）1 [导读] 海南医学院附属医院神经内科贵阳中医学院研究生院在读硕士随着神经科学、药理学的不断发展，相信可能有更好的方法缓解或者治愈帕金森病运动并发症。
1.海南医学院附属医院神经内科海南海口 5700002.贵阳中医学院研究生院在读硕士贵州贵阳 550002 摘要：目的：探讨咖啡因在帕金森运动并发症的应用方法：广泛查阅国内外相关咖啡因与帕金森运动并发症关系的资料，并进行分析和总结。
结果：咖啡因有助于改善帕金森病患者的运动能力。
结论：随着神经科学、药理学的不断发展，相信可能有更好的方法缓解或者治愈帕金森病运动并发症。
关键词：帕金森病；运动并发症；咖啡因PD是目前第二个最为常见的一种神经退行性疾病，其病理特征是基底神经节递质紊乱，导致了PD症状的出现。
治疗上公认为L-DOPA 治疗PD仍是最有效的药物，但是随着病情进展，它会引发运动并发症。
本文就咖啡因的功能、在神经系统中的功能以及帕金森病运动并发症的实验应用进行综述，对PD的病情发展以及预防和治疗策略方面提供一定的参考，给PD的预防和治疗提供新思路。
1、咖啡因的概述1.1咖啡因的功能咖啡因为腺苷2A受体拮抗剂，在人体的多个系统中具有广泛的药理学作用，它会刺激中枢神经系统、心脏和呼吸的节律，对精神属性的改变。
1.2 咖啡因在神经系统中功能咖啡因具有降低老年痴呆症、阿尔茨海默氏病、PD等发病率。
咖啡因可以调控学习和记忆功能，还能提高认知等功能并阻止年龄相关记忆编码的损失。
2、咖啡因在PD中的表达及应用咖啡因在信号通路中由于腺苷拮抗作用而导致许多下游的变化，包括多巴胺能系统的变化、多巴胺受体和基因表达亲和力降低，咖啡因正是通过这些机制来介导其对大脑和行为的影响。
咖啡因通过阻断腺苷A2A受体来保护神经元，但是咖啡因对PD的风险呈负相关还只是一个微弱的证据。
白藜芦醇对D_半乳糖致衰老小鼠学习记忆能力和脑组织抗氧化能力的影响
Effects of Resveratrol on Learning, Memory and Brain Antioxidan t Abilities in D -Galactose-Induced Aging Mice白藜芦醇对D - 半乳糖致衰老小鼠学习记忆能力和 脑组织抗氧化能力的影响刘贵珊1,2，杨 博3，张泽生2,*，范艳丽1，贺 伟2（1.宁夏大学农学院，宁夏 银川750021；2.天津科技大学食品工程与生物技术学院，天津 300457； 3.宁夏大学新华学院，宁夏 银川750021）摘 要：探讨白藜芦醇对D -半乳糖致衰老小鼠脑组织抗氧化与学习记忆的影响，揭示白藜芦醇延缓脑衰老的作用。
将50 只雄性ICR 鼠随机分为正常对照组，模型组，白藜芦醇低、中、高剂量组。
模型组及白藜芦醇治疗组连续8 周于小鼠颈背部皮下注射D -半乳糖120 mg/（kg ·d ），给予不同剂量白藜芦醇（25、50、100 mg/（kg ·d ））灌 胃；正常对照组注射、灌胃等量生理盐水。
采用Morris 水迷宫检测各组小鼠的学习记忆能力，并测定各组小鼠脑 组织超氧化物歧化酶（superoxide dismutase ，SOD ）、总抗氧化能力（total antioxidant capacity ，T-AOC ）、丙二 醛（malondialdehyde ，MDA ）的含量。
白藜芦醇能够显著缩短D -半乳糖致衰老小鼠Morris 水迷宫中找到隐藏平台 时间（P ＜0.01），提高游泳速度（P ＜0.01）和穿越目标象限的次数（P ＜0.01）；提高衰老模型小鼠脑组织SOD（P ＜0.01）、T-AOC 活性（P ＜0.05），降低MDA 含量（P ＜0.01）。
白藜芦醇能够改善D -半乳糖致衰老模型小鼠 的学习记忆能力，提高脑组织的抗氧化能力，具有一定延缓脑衰老的作用。
关键词：白藜芦醇；D -半乳糖；学习记忆能力；抗氧化LIU Gui-shan 1,2, Y ANG Bo 3, ZHANG Ze-sheng 2,*, FAN Y an-li 1, HE Wei 2(1. School o f Agriculture, Ningxia University , Yinchuan 750021, China; 2. College of Food Engineering and Biotechnology, Tianj in University of Science and Technology, Tianjin 300457, China; 3. Xinhua College, Ningxia University, Yinchuan 750021, China)Abstract: Objective: To investigate the effects of resveratrol (Res ) on learning, memory a nd brain anti oxidant capacitiesin D -galactose-induced a ging mice. Methods: Fifty ICR mice were randomly divided i nto five groups, designated a s n ormal control, model, resveratrol low-dose, moderate-dose and high-dose groups. The normal control group was given normal saline by g avage, and all other groups were given D -galactose solution by n eck back subcutaneous injection at the daily dose of 120 mg/(kg·d ) for 8 consecutive weeks. At the same time , Res-treated mice were administered Res by gavage at the daily doses of 25, 50 and 100 mg/(kg·d ) body weight per day, respectively. The learning and m em ory abilities of mice were determined by Morris water maze test. Superoxide dismutase (SOD), total antioxidant capacity (T-AOC) and malondialdehyde (MDA) i n t he b rain o f mice w ere d etermined at t he e nd o f 12 h o f fasting a fter the l ast a dministration. Results: Res c ould reduce the time mice spent on finding platform (P ＜ 0.01), improve the sw imming speed (P ＜ 0.01) and the number o f times mice crossed the platform (P ＜ 0.01) in Morris water maze test, increase SOD (P ＜ 0.01) and T-AOC activities (P ＜ 0.05) and decrease the content of MDA (P ＜ 0.01) in brain tissue. Conclusion: Res c an ameliorate the degeneration of learning an d memory abilities in D -galactose-induced aging mice, improve brain antioxidant abilities and thus delay brain aging. Key words: resveratrol; D -galactose; learning and m em ory abilities ; antioxidation 中图分类号：TS201.2文献标志码：A 文章编号：1002-6630（2014）05-0204-04 doi:10.7506/spkx1002-6630-201405040随着人类平均预期寿命的不断延长，世界人口结构 的老龄化趋势日益突显，一个人口老龄化的时代已经来临[1]。
超氧阴离子对牛主动脉平滑肌细胞收缩性的影响
超氧阴离子对牛主动脉平滑肌细胞收缩性的影响承伟;小山澈野;大池正宏【期刊名称】《中国药理学通报》【年(卷),期】2012(28)9【摘要】Aim To investigate the changes of [ Ca2 + ]; and contractility of aortic smooth muscle cells induced by superoxide anion. Methods The aortic smooth muscle cells were primarily cultured. The effect of superoxide anion on [ Ca2+ ]; was measured by a digital fluorescence microscopy system. Muscle contractility was analyzed by collagen gel contraction system. Results High-K+ and A-23187-induced [ Ca2+ ]; elevated both in control and X/XO-treated cells. The net increment of [ Ca2 + ]; was not significantly different between control and X/XO-treatedcells( P >0. 05 ). Col-rnlagen gel contraction area in X/XO-treated cells induced by High-K+ and A-23187 was impaired from 18. 32% ±2. 11% to 5. 13% ±0. 45%( P <0. 01 )and 17. 16% ± 1. 10% to 4. 31% ± 0. 04% ( P < 0. 01 ). Conclusion The results demonstrate that superoxide anion attenuates smooth muscle contraction, whereas [ Ca2+ ]; increased by these agents were not affected.%目的观察超氧阴离子对培养的牛动脉平滑肌细胞内钙及收缩性的影响.方法原代培养牛动脉平滑肌,使用数字荧光显微镜技术测定细胞内钙,采用胶原凝胶收缩方法分析收缩性.结果超氧阴离子对高钾溶液及A-23187诱导的钙内流无明显影响(P>0.05),但可使凝胶收缩面积减小.超氧阴离子可使高钾诱导的胶原凝胶收缩面积从18.32%±2.11%减小至5.13%±0.45%(P<0.01),使A-23187诱导的胶原凝胶收缩面积从17.16%±1.10%减小至4.31%±0.04%(P<0.01).结论超氧阴离子可明显抑制牛主动脉平滑肌细胞收缩性,但对电压依赖性钙通道引起的钙内流及A-23187诱导的细胞内钙变化无影响.【总页数】3页(P1215-1217)【作者】承伟;小山澈野;大池正宏【作者单位】辽宁医学院药学院,辽宁,锦州,121000;九州大学药理学教研室,日本,福冈,812-8512;九州大学药理学教研室,日本,福冈,812-8512【正文语种】中文【中图分类】R-332;R322.121;R322.74;R348.1;R977.9【相关文献】1.超声辐照对血管紧张素Ⅱ诱导的牛主动脉平滑肌细胞增殖的影响 [J], 李妍妍;徐标;吴巍;冯若2.超氧阴离子对培养的牛动脉平滑肌细胞内钙及收缩性的影响 [J], 承伟;李智;小山澈野;大池正宏;伊东裕之3.3,6-(二甲氨基)-二苯骈碘杂六环葡萄糖酸盐对AGEP引起的大鼠主动脉平滑肌细胞增殖及牛主动脉内皮细胞内皮素和一氧化氮改变的影响 [J], 邓秀玲;刘乃丰;钱之玉;侯自杰4.苯扎贝特对培养的牛主动脉血管平滑肌细胞增殖的影响 [J], 赵荫涛; 赵春霞; 汪培华; 汪道文5.牛主动脉蛋白聚糖对培养的人主动脉平滑肌细胞生长的影响 [J], 丛祥凤;刘学文;张英珊因版权原因，仅展示原文概要，查看原文内容请购买。
莫诺苷对小鼠骨髓源树突状细胞表型及功能的影响
莫诺苷对小鼠骨髓源树突状细胞表型及功能的影响李慈;姚雪;夏圣;张燕;周婧文;仇欣;吴仪;赵传祥;高凤威;周文慧;刘烁【摘要】目的:探究莫诺苷对小鼠骨髓源树突状细胞(bone-marrow derived dendritic cells,BMDCs)分化、表型和功能的影响.方法:体外培养C57BL/6小鼠BMDCs,分为对照组(RPMI 1640)、莫诺苷组、脂多糖组和莫诺苷+脂多糖联合处理组.用光学显微镜以及流式细胞术分别确认BMDCs形态变化和羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)的荧光强度;流式细胞术检测BMDCs表面CD80、CD86、MHC-Ⅰ、MHC-Ⅱ类分子的表达情况以及激活抗原特异性CD4+T细胞活化、增殖和分化的能力;ELISA法检测BMDCs培养上清液中细胞因子IL-6和IL-12的含量.结果:与对照组相比,莫诺苷处理后BMDCs诱导分化过程中的增殖效率无明显差异;与脂多糖组相比,莫诺苷+脂多糖联合处理后上调成熟BMDCs表面CD80、CD86、MHC-Ⅰ、MHC-Ⅱ类分子的表达和IL-12的分泌,并促进BMDCs激活OVA323-339抗原特异性T细胞的活化、增殖以及向Th1分化.结论:莫诺苷促进小鼠BMDCs的成熟,上调其对OVA特异性CD4+T细胞的抗原提呈功能.【期刊名称】《江苏大学学报（医学版）》【年(卷),期】2019(029)004【总页数】5页(P277-281)【关键词】莫诺苷;树突状细胞;共刺激分子;CD4+T细胞【作者】李慈;姚雪;夏圣;张燕;周婧文;仇欣;吴仪;赵传祥;高凤威;周文慧;刘烁【作者单位】江苏大学医学院,江苏镇江212013;江苏大学医学院,江苏镇江212013;江苏大学医学院,江苏镇江212013;江苏大学医学院,江苏镇江212013;江苏大学医学院,江苏镇江212013;江苏大学医学院,江苏镇江212013;江苏大学医学院,江苏镇江212013;江苏大学医学院,江苏镇江212013;江苏大学医学院,江苏镇江212013;江苏大学医学院,江苏镇江212013;江苏大学医学院,江苏镇江212013【正文语种】中文【中图分类】R392.12莫诺苷（morroniside）是一种从山茱萸中提取的物质，可以促进神经功能的恢复［1-3］。
桑白皮多酚对B16细胞内黑色素生成的影响及其机制
桑白皮多酚对B16细胞内黑色素生成的影响及其机制吴永祥;毕淑峰;姜薇;崔谱;金有贞;金泰完【摘要】Aim To investigate the inhibitory effect of polyphenol from Cortex Mori( CMP) on melanogenesis in mouse melanoma B16 cells and its possible mecha- nism. Methods Melanoma B16 cells with high ex-pression melanin were induced by α-melanocyte-stimu-lating hormone ( α-MSH) to establish cell model. Cell viability was detected by MTT assay. The melanin syn-thesis and tyrosinase activity were measured by NaOH and L-Dopa assays, respectively. The tyrosinase (TYR), tyrosinase-related protein-1 (TRP-1), tyrosi-nase-related protein-2 ( TRP-2 ) and microphthalmia associated transcription factor ( MITF ) protein and mRNA levels were measured by Western blot and qRT-PCR, respectively. Results CMP could inhibit the melan in synthesis and tyrosinase activity in α-MSH stimulated B16 cells in a dose-dependent manner ( P<0.05) . The melanin content and tyrosinase activity significantly decreased by 52.95% , 32.85% at 20 mg ·L-1of CMP, respectively. Treatment of 100 mg· L-1of arbutin reduced the melanin content and tyrosi- nase activity by 17.29% , 16.75% , respectively. Based on the results of this study, CMP showed a stronger anti-melanogenesis activity than that of positive control arbutin. After treated by CMP, the protein and mRNA levels of TYR, TRP-1, TRP-2 and MITF were significantly inhibited compared to the α-MSH group ( P<0.05) . Conclusions CMP could suppress the melanogenesis in α-MSH stimulated B16 cells, and its mechanism may be related to its regulation of the pro-tein and mRNA expressions of TYR, TRP-1, TRP-2 and MITF, and the inhibition of tyrosinase activity.%目的研究桑白皮多酚( polyphenol from Cortex Mori, CMP)对小鼠B16细胞内黑色素生成的影响,并探究其作用机制.方法体外培养小鼠B16 细胞,构建α-黑素细胞刺激素( α-MSH)诱导的黑色素高表达细胞模型. CMP 干预B16细胞,MTT法测定细胞活性;分别利用NaOH裂解法和L-Dopa氧化法,分析细胞内黑色素生成含量和酪氨酸酶活性的变化;Western blot 和实时荧光定量 PCR 法分别测定B16细胞中酪氨酸酶(TYR)、酪氨酸酶相关蛋白-1(TRP-1)、酪氨酸酶相关蛋白-2(TRP-2)、小眼畸形相关转录因子(MITF)的蛋白质和mRNA水平.结果 CMP对α-MSH诱导的B16细胞内黑色素生成及酪氨酸酶活力均具有明显的抑制作用(P<0.05),且呈量效关系.当CMP浓度为20 mg ·L-1时,对细胞内黑色素生成及酪氨酸酶活性抑制率分别为52.95% 、32.85% ,阳性对照熊果苷(100 mg·L-1)的抑制率分别为17.29% 、16.75% ,表明CMP对黑色素生成的抑制效果强于熊果苷.与α-MSH模型组相比,CMP干预后细胞内TYR、TRP-1、TRP-2、MITF的mRNA和蛋白表达被明显抑制(P<0.05).结论 CMP明显抑制α-MSH诱导黑色素的生成,其机制可能是通过调控 TYR、TRP-1、TRP-2、MITF mRNA和蛋白表达,进而抑制酪氨酸酶活性实现的.【期刊名称】《中国药理学通报》【年(卷),期】2018(034)009【总页数】6页(P1296-1301)【关键词】桑白皮;多酚;B16 细胞;α-黑素细胞刺激素;黑色素;酪氨酸酶【作者】吴永祥;毕淑峰;姜薇;崔谱;金有贞;金泰完【作者单位】黄山学院生命与环境科学学院,安徽黄山245041;安东国立大学食品科学与生物技术学院,韩国安东 760749 ;黄山学院生命与环境科学学院,安徽黄山245041;黄山学院生命与环境科学学院,安徽黄山245041;黄山学院生命与环境科学学院,安徽黄山245041;安东国立大学食品科学与生物技术学院,韩国安东760749 ;安东国立大学食品科学与生物技术学院,韩国安东 760749【正文语种】中文【中图分类】R-332;R284.1;R329.24;R345.49;R348.6;R739.5;R977.3;R977.7黑色素是由黑色素细胞合成的高分子生物色素，分布于皮肤真皮层中，决定了皮肤的颜色。
细菌在氨基酸首过肠道代谢中的作用
细菌在氨基酸首过肠道代谢中的作用杨宇翔;慕春龙;朱伟云【摘要】Intestinal bacteria play important roles in nutrient metabolism, which furtherly affect body metabo-lism. Small intestine is the main region for amnio acids metabolism, and is critical to the host protein nutrition and gut health. However, large amount of amino acids entered into small intestine was degraded during the first-pass intestinal metabolism by bacteria. Amino acid metabolism in the small intestine is compartmentalized, not only segmented, but also layered. Amino acid metabolites, such as polyamine and dipeptide, are important for host gut health. Moreover, urea degradation and subsequent amino acid synthesis by the bacteria might be nutritionally valuable for the host. This article mainly reviewed the roles and out-ways in the first-pass intestinal metabolism of amino acids, metabolic compartments of amino acids, and the impacts of the metabolites on host health of small intestinal bacteria.%肠道细菌在营养素代谢过程中起重要作用，进而影响机体整体代谢。