护卵汤对慢性应激超排卵小鼠卵巢TGF-β/Smads信号通路蛋白的影响
作者：
刘碧源
（ ;申可佳
（ ;申奏秦旋;付灵梅;熊桀
期刊：
中华中医药杂志 ,2018年33(8):3332-3335 ISSN：1673-1727
作者机构：
湖南中医药大学医学院, 长沙, 410208;湖南中医药大学第一附属医院, 长沙, 410208;[申可佳; 刘碧源; 申奏秦旋; 熊桀] 湖南中医药大学医学院, 长沙, 410208;[付灵梅] 湖南中医药大学第一附属医院, 长沙, 410208
关键词：
护卵汤;慢性应激;超排卵;转化生长因子β1;卵泡刺激素受体
摘要：
目的:观察护卵汤对慢性应激超排卵小鼠卵巢转化生长因子β(TGF-β)/Smads信号通路相关蛋白表达的影响。方法:建立慢性应激小鼠模型,将模型小鼠随机分为中药(护卵汤)组、西药(生长激素+阿司匹林)组和模型组,并设正常组。模型组和正常组给予0.9%氯化钠溶液,治疗组给予相应的药物。在超排卵后第3天处死小鼠10只;注射绒毛膜促性腺激素(HCG)24h后处死余下小鼠取卵巢。Western Blot检测卵巢TGF-β1、 P-Smad3及卵泡刺激素受体(FSHR)蛋白表达。结果:模型组卵巢TGF-β1、P-Smad3及FSHR蛋白表达较正常组同期均显著降低(P<0.05)。与模型组同期比较,中药组和西药组卵巢TGF-β1、P-Smad3及FSHR蛋白表达均明显升高(P<0.05);且中药组与西药组同期比较升高更明显(P<0.05)。结论:护卵汤可促进慢性应激小鼠卵巢功能的恢复,其机制可能与调控TGF-β/Smads信号通路相关蛋白表达有关。
语种：
中文
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医学免疫学系统性教学的实践
期刊：
山西医科大学学报 ,2017年19(2):85-87 ISSN：1007-6611
作者机构：
湖南中医药大学医学院免疫学教研室,长沙,410208;[陈超龙; 伍参荣; 申可佳; 刘碧源] 湖南中医药大学
关键词：
医学免疫学;教学改革;教学方法;系统性教学
摘要：
教学要遵循系统性原则,文章就如何在本科医学免疫学课程中进行系统性教学进行了详细阐述。首先,要求教师把握教学内容的系统性,为学生总结出该学科知识体系的主线;其次,结合学生专业特点,为学生构建起医学免疫学学习所需的系统性知识结构;再次,探索了医学免疫学理论联系实践的系统性教学。系统性教学提高了医学生对医学免疫学课程的学习效果,为临床医学人才的培养打下了基础。
语种：
中文
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临床医学专业医学免疫学课程说课设计的理念
期刊：
山西医科大学学报 ,2017年19(12):932-935 ISSN：1007-6611
作者机构：
湖南中医药大学医学院免疫学教研室,长沙,410208;[陈超龙; 伍参荣; 申可佳; 刘碧源] 湖南中医药大学
关键词：
医学免疫学;说课;临床医学专业;教学改革
摘要：
医学免疫学是临床医学专业的一门专业基础课程,对学生今后临床课程的学习和临床岗位工作能力的培养具有重要的作用。医学免疫学又是一门理论性和实践性较强的学科,内容错综复杂,教和学都有一定的难度。为了促进该课程建设,文章从课程定位与目标、课程设计与教学内容选取、教学组织与实施、教学方法与手段、课程特色五个方面对临床医学专业医学免疫学课程进行说课设计。
语种：
中文
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课前预习加小讲课在医学免疫学教学中的应用
期刊：
卫生职业教育 ,2017年35(7):53-54 ISSN：1671-1246
作者机构：
湖南中医药大学医学院,湖南 长沙,410208;[陈超龙; 伍参荣; 申可佳; 刘碧源] 湖南中医药大学
关键词：
医学免疫学;课前预习;小讲课
摘要：
医学免疫学是基础医学中最抽象、难懂的一门学科,如何提高课堂教学效果和教学质量,一直是教学改革的方向。课前预习可有效提高教学效果和质量,但在大学教育中由于没有相应的督促措施与评价手段,学生预习效果往往并不理想。将课前预习与小讲课相结合应用于医学免疫学教学,有利于保证知识的系统性和连贯性,从而提高教学质量。
语种：
中文
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医学免疫学实验课中加强生物安全教育重要性的探讨
期刊：
山西医科大学学报 ,2016年18(7):544-546 ISSN：1007-6611
作者机构：
湖南中医药大学医学院免疫学教研室,长沙,410208;[陈超龙; 伍参荣; 申可佳; 刘碧源] 湖南中医药大学
关键词：
医学免疫学;实验教学;生物安全教育
摘要：
医学免疫学实验课离不开病原微生物,加强生物安全教育非常必要。文章分析了医学免疫学实验教学中主要存在的生物安全问题,根据实验课的自身特点,从树立学生的生物安全意识、加强实验前的准备、实验中的反复强调、实验后的恰当处置和意外情况及时处理等方面,切实保证学生掌握标准的实验操作技术,养成良好的职业行为,确保学生实验室的生物安全。
语种：
中文
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Identification of phage display peptides with affinity for the tegument of Schistosoma japonicum schistosomula
作者：
Liu, Yan
（ ;Brindley, Paul J.;Zeng, Qingren;Li, Yuesheng;Zhou, Jun;...
期刊：
Molecular and Biochemical Parasitology ,2011年180(2):86-98 ISSN：0166-6851
通讯作者：
Zeng, QR
作者机构：
[Yang, Shenghui; Liu, Yan; Zhang, Zuping; Cai, Liting; Zeng, Qingren; Chen, Yuxiao; Li, Yuesheng] Cent S Univ, Xiangya Sch Med, Ctr Cell & Mol Biol Expt, Changsha 410013, Hunan, Peoples R China.;[Brindley, Paul J.] George Washington Univ, Med Ctr, Dept Microbiol Immunol & Trop Med, Washington, DC 20037 USA.;[McManus, Donald P.; Li, Yuesheng] Queensland Inst Med Res, Mol Parasitol Lab, Brisbane, Qld 4006, Australia.;[Liu, Yan] Univ S China, Sch Med, Dept Parasitol, Hengyang 421001, Peoples R China.;[Zhou, Jun] Cent S Univ, Xiangya Hosp 3, Expt Ctr, Changsha 410013, Hunan, Peoples R China.
通讯机构：
[Zeng, QR ] C;Cent S Univ, Xiangya Sch Med, Ctr Cell & Mol Biol Expt, Tongzipo Rd 172, Changsha 410013, Hunan, Peoples R China.
关键词：
helminth protein;peptide library;peptide ZL4;polypeptide;rhodamine B;unclassified drug;absorption;article;binding affinity;biotransformation;conjugation;controlled study;developmental stage;host parasite interaction;immunoblotting;immunohistochemistry;in vitro study;in vivo study;integument;nonhuman;parasite identification;peptide synthesis;phage display;priority journal;protein binding;protein isolation;protein localization;real time polymerase chain reaction;Schistosoma japonicum;Schistosomulum;surface property;trematode life cycle stage;Animals;Female;Humans;Kinetics;Mice;Peptide Library;Peptides;Protein Binding;Schistosoma japonicum;Schistosomiasis japonica;Mammalia;Schistosoma;Schistosoma japonicum
摘要：
Peptides, bound to the tegument of live Schistosoma japonicum schistosomula, were differentially screened by phage display in vitro using three rounds of reverse absorption and bio-panning. Three M13 phage peptides were isolated and identified by determination of their recovery rate, immunohistochemical localization, immunoblot analysis, and their anti-schistosomal effects in vivo and in vitro. Of the three, M13 phage peptide ZL4 (MppZL4, YSGLQDSSLRLR, 1.4 kDa, pI 8.8) bound to the tegument of mechanically transformed schistosomula and to other developmental stages of S. japonicum from the mammalian host. By contrast, MppZL4 did not bind to the surface of cercariae. To further examine its binding properties, MppZL4 was conjugated to Rhodamine B (RhB-YSGLQDSSLRLR, RhB-ZL4) and a peptide control (RhB-AIPYFSGILQWR, RhB-12P) was similarly synthesized. The binding capacities of RhB-ZL4 to the surface membrane of S. japonicum schistosomula in vitro and of S. japonicum adult worms in vivo were examined and revealed specificity for binding. When examined for anti-parasite activity, both MppZL4 and RhB-ZL4 exhibited a potent schistosomicidal effect in vitro. Further MppZL4 also affected the growth and development of schistosomula in vivo. These findings extend previous studies showing that phage display techniques can recover polypeptides that bind specifically to living schistosomes and, moreover, that these bound peptides have the potential to inhibit key physiological processes in these parasites. Our findings suggest further that ectogenic polypeptides, which can bind to the tegument of S. japonicum, might be adapted as vectors to deliver experimental probes and/or pharmacologically relevant compounds to the schistosome tegument, including drugs and immunological mediators. © 2011 2011 Elsevier B.V. All rights reserved.
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英文
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SV40LT基因重组腺病毒载体的包装、鉴定及用于转染日本血吸虫童虫细胞的研究
作者：
刘碧源
（ ;曾庆仁;余平;杨胜辉
（ ;蔡力汀;...
期刊：
中国人兽共患病学报 ,2011年27(11):1016-1020 ISSN：1002-2694
作者机构：
[刘碧源; 曾庆仁; 余平; 蔡力汀; 张顺科] 中南大学湘雅医学院细胞与分子生物学实验中心;南华大学公共卫生学院卫生检验系;[杨胜辉] 湖南中医药大学病原生物学与免疫学教研室;[周军] 中南大学湘雅三医院实验中心
关键词：
SV40LT基因;腺病毒载体;日本血吸虫;童虫细胞
摘要：
目的 构建携带SV40LT基因的重组腺病毒表达载体，制备具有感染力的重组腺病毒，观察其转染日本血吸虫（Schistosoma japonicum,Sj）童虫细胞后的表达情况。方法 采用体外连接法构建好的携带SV40LT基因的重组腺病毒质粒(AdHu5-SV40LT)转化Stb12感受态菌，获得重组腺病毒质粒后，经Pac I酶切线性化后转染293A细胞，获得出重组腺病毒（AdHu5-SV40LT）。将重组腺病毒感染Sj童虫细胞，采用RT-PCR和免疫组织化学检测SV40LT基因的表达情况。结果 重组腺病毒载体质粒可转染293A细胞并可在293A细胞内进行有效的复制；提取病毒DNA进行PCR检测证实含有 SV40LT目的基因。以重组腺病毒能感染Sj童虫细胞，经RT-PCR和免疫组化检测有SV40LT基因在细胞中的表达。结论 成功包装了具有感染能力的含SV40LT基因的重组腺病毒，感染Sj童虫细胞后目的基因有表达，为进一步探索日本血吸虫细胞永生化提供了实验依据。
语种：
中文
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Pantropic retroviral vectors pseudotyped with the glycoprotein of vesicular stomatitis virus (VSVG) mediate gene transfer and transgene expression in Schistosoma japonicum schistosomules
作者：
Yang, S. H.
（ ;Brindley, P. J.;Zeng, Q. R.;Zeng, T. B.;Li, Y. S.;...
作者机构：
[Liu, Y.; Yang, S. H.; Liu, B. Y.; Li, Y. S.; Cai, L. T.; Zeng, Q. R.; Zeng, T. B.; Lan, L. M.] Cent S Univ, Xiangya Sch Med, Ctr Cell & Mol Biol Expt, Changsha, Peoples R China.;[Yang, S. H.; Wu, C. R.; Tan, Z. J.; Lu, F. G.] Hunan Univ Chinese Med, Dept Pathogen Biol & Immunol, Changsha, Peoples R China.;[Brindley, P. J.] George Washington Univ, Med Ctr, Dept Microbiol Immunol & Trop Med, Washington, DC 20037 USA.;[Liu, Y.; Liu, B. Y.; Zeng, T. B.] Univ South China, Sch Med, Hengyang, Peoples R China.;[Li, Y. S.; McManus, D. P.] Queensland Inst Med Res, Mol Parasitol Lab, Brisbane, Qld, Australia.
会议名称：
12th International Congress of Parasitology (ICOPA)/6th Novel Approaches to the Control of Helminth Parasites of Liverstock Conference
会议时间：
AUG 15-20, 2010
会议地点：
Melbourne, AUSTRALIA
会议主办单位：
[Yang, S. H.;Zeng, Q. R.;Zeng, T. B.;Li, Y. S.;Liu, Y.;Liu, B. Y.;Cai, L. T.;Lan, L. M.] Cent S Univ, Xiangya Sch Med, Ctr Cell & Mol Biol Expt, Changsha, Peoples R China.^[Yang, S. H.;Wu, C. R.;Lu, F. G.;Tan, Z. J.] Hunan Univ Chinese Med, Dept Pathogen Biol & Immunol, Changsha, Peoples R China.^[Brindley, P. J.] George Washington Univ, Med Ctr, Dept Microbiol Immunol & Trop Med, Washington, DC 20037 USA.^[Zeng, T. B.;Liu, Y.;Liu, B. Y.] Univ South China, Sch Med, Hengyang, Peoples R China.^[Li, Y. S.;McManus, D. P.] Queensland Inst Med Res, Mol Parasitol Lab, Brisbane, Qld, Australia.^[Zhou, J.] Cent S Univ, Expt Ctr Xiangya Third Hosp, Changsha, Hunan, Peoples R China.
摘要：
Retroviral transduction of cultured schistosomes was explored in our study. The vesicular stomatitis virus glycoprotein (VSVG)-pseudotyped pantropic retroviral vector pBABE-puro was modified to incorporate the human telomerase reverse transcriptase gene (hTERT) as a reporter gene under the control of retroviral long terminal repeat (LTR). Pseudotyped virions were employed to transduce Schistosoma japonicum (Sj) to investigate the utility of retrovirus mediated Sj transgenesis and to investigate the activity as human telomerase as a reporter transgene in schistosomes. The presence of transgene hTERT was detected by PCR. Furthermore, analysis of RNAs from transduced parasites, immunohistochemistry of thin sections and immunoblot analysis revealed expression of hTERT transgene in the transduced worms. These findings indicated that Sj could be effectively transduced by VSVG pseudotyped retrovirus carrying the hTERT gene.
语种：
英文
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Transduction of Schistosoma japonicum schistosomules with vesicular stomatitis virus glycoprotein pseudotyped murine leukemia retrovirus and expression of reporter human telomerase reverse transcriptase in the transgenic schistosomes
作者：
Yang, Shenghui
（ ;Brindley, Paul J.;Zeng, Qingren;Li, Yuesheng;Zhou, Jun;...
期刊：
Molecular and Biochemical Parasitology ,2010年174(2):109-116 ISSN：0166-6851
通讯作者：
Zeng, QR
作者机构：
[Yang, Shenghui; Liu, Yan; Cai, Liting; Li, Yuesheng; Zeng, Tiebing; Lan, Lingmei; Wei, Qi; Zeng, Qingren] Cent South Univ, Xiangya Sch Med, Ctr Cell & Mol Biol Expt, Changsha 410013, Hunan, Peoples R China.;[Brindley, Paul J.] George Washington Univ, Med Ctr, Dept Microbiol Immunol &Trop Med, Washington, DC 20037 USA.;[McManus, Donald P.; Li, Yuesheng] Queensland Inst Med Res, Mol Parasitol Lab, Brisbane, Qld 4006, Australia.;[Yang, Shenghui] Hunan Univ Chinese Med, Dept Pathogen Biol & Immunol, Changsha 410208, Hunan, Peoples R China.;[Zhou, Jun] Cent South Univ, Xiangya Hosp 3, Expt Ctr, Changsha 410013, Hunan, Peoples R China.
通讯机构：
[Zeng, QR ] C;Cent South Univ, Xiangya Sch Med, Ctr Cell & Mol Biol Expt, 172 Tongzipo Rd, Changsha 410013, Hunan, Peoples R China.
关键词：
genomic DNA;RNA;telomerase reverse transcriptase;virus glycoprotein;article;cell growth;cell proliferation;gene expression;host parasite interaction;immunohistochemistry;long terminal repeat;nonhuman;parasite;priority journal;reporter gene;Retrovirus;reverse transcription polymerase chain reaction;Schistosoma;Schistosoma japonicum;schistosomiasis;transgene;transgenics;Vesicular stomatitis virus;virion;virus detection;Animals;Animals, Genetically Modified;Cell Line;Genes, Reporter;Humans;Immunoblotting;Immunohistochemistry;Leukemia Virus, Murine;Membrane Glycoproteins;Mice;NIH 3T3 Cells;Polymerase Chain Reaction;Rabbits;Reverse Transcriptase Polymerase Chain Reaction;Schistosoma japonicum;Telomerase;Transduction, Genetic;Transgenes;Viral Envelope Proteins;Murinae;Murine leukemia virus;Oryctolagus cuniculus;Schistosoma;Schistosoma japonicum;Schistosoma mansoni;Vesicular stomatitis virus
摘要：
Although draft genome sequences of two of the major human schistosomes, Schistosoma japonicum and Schistosoma mansoni are available, the structures and characteristics of most genes and the influence of exogenous genes on the metabolism of schistosomes remain uncharacterized. Furthermore, which functional genomics approaches will be tractable for schistosomes are not yet apparent. Here, the vesicular stomatitis virus glycoprotein (VSVG)-pseudotyped pantropic retroviral vector pBABE-puro was modified to incorporate the human telomerase reverse transcriptase gene (hTERT) as a reporter, under the control of the retroviral long terminal repeat (LTR). Pseudotyped virions were employed to transduce S. japonicum to investigate the utility of retrovirus-mediated transgenesis of S. japonicum and the activity of human telomerase reverse transcriptase as a reporter transgene in schistosomes. Schistosomules perfused from experimentally infected rabbits were cultured for 6 days after exposure to the virions after which genomic DNAs from virus exposed and control worms were extracted. Analysis of RNA from transduced parasites and immunohistochemistry of thin parasite sections revealed expression of hTERT in the transduced worms. Expression of hTERT was also confirmed by immunoblot analysis. These findings indicated that S. japonicum could be effectively transduced by VSVG-pseudotyped retrovirus carrying the hTERT gene. Given the potential of hTERT to aid in derivation of immortalized cells, these findings suggest that this pantropic retroviral approach can be employed to transduce cells from specific tissues and organs of schistosomes to investigate the influence of transgene hTERT on growth and proliferation of schistosome cells. © 2010 Elsevier B.V.
语种：
英文
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双嗜性逆转录病毒感染日本血吸虫细胞的生物学理论与可行性探讨
作者：
杨胜辉
（ ;秦志强;曾庆仁;曾铁兵;刘彦
（ ;...
期刊：
中国病原生物学杂志 ,2009年4(10): 744-753,776,封3 ISSN：1673-5234
作者机构：
中南大学湘雅医学院寄生虫学系,湖南长沙410013;湖南中医药大学病原生物学与免疫学教研室,湖南长沙410208;中南大学湘雅医学院寄生虫学系,湖南长沙,410013;南华大学病原生物学研究所,湖南衡阳421001;[杨胜辉] 湖南中医药大学
关键词：
双嗜性逆转录病毒;受体同源蛋白;血吸虫;日本;培养细胞;基因整合与表达
摘要：
目的 探讨用双嗜性逆转录病毒载体将外源基因导入日本血吸虫(Sj)细胞的生物学理论与实验依据. 方法 用生物信息学方法对双嗜性逆转录病毒rRam-1受体同源性分布、结构与功能作系统的分析与比较;利用携带外源E77.43基因的双嗜性逆转录病毒感染Sj童虫培养细胞,经PCR和RT-PCR检测感染细胞目的 基因(E77.43)的整合与表达. 结果 根据生物信息学分析结果推断,Sj细胞膜上存在的SiCHGC09605和SjCHGC05362两种蛋白为非分泌性跨膜蛋白,可能具有细胞膜离子转运通道或受体蛋白的功能及双嗜性逆转录病毒感染的膜受体样作用,可能参与病毒对细胞的吸附和穿入过程;利用携带外源E77.43基因的双嗜性逆转录病毒感染Sj童虫培养细胞后.用PCR及RT-PCR检测到目的 基因整合与表达,扩增的目的 片段大小为330 bp,与理论值相符. 结论 用载有E77.43基因的双嗜性逆转录病毒感染Sj童虫细胞获得成功,推测SiCHGC09605和SjCHGC05362两种与rRam-1受体同源的蛋白可能是Sj感染过程中起作用的分子.研究结果为下一步用双嗜性逆转录病毒载体转导永生化基因到Sj细胞提供了生物学理论与实验依据.
语种：
中文
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