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Observations of Nuclear Status in Canine Oocytes Immediately after Collection from Ovaries Reveal Three Types of Germinal Vesicle

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Observations of Nuclear Status in Canine Oocytes Immediately...

CANINE OOCYTES CAN BE SPONTANEOUSLY PARTHENOGENERATED AND DEVELOP TO 2-CELL TO 8-CELL STAGE PARTHENOGENERATED EMBRYOS IN COCULTURE SYSTEM WHICH THEY WERE COCULTURED WITH CANINE OVIDUCTAL CELLS OR CANINE OVIDUCT EXTRACTS.

全国动物生理生化第十次学术交流会

作者： Lunxue Wang~1 Hong Guan~1 Rongliang Hu~2 Xiaorong An~(1*) 1 State Key Laboratory of Agrobiotechnology,Department of Biological sciences,China Agricultural University,Beijing 100094,China 2 The Insititue of Veterinary Medicine,academy of military medical science of pla,Changchun 130062,China

The objective of this study was to assess the stage of meiotic maturation in canine oocytes without in vitro culture at different season and examine whether varieties of GVs observed in this investigation associated w...

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CANINE OOCYTES CAN BE SPONTANEOUSLY PARTHENOGENERATED AND DE...

The 5th Annual Conference of the Asian Reproductive Biotechnology Society

作者： Lunxue WANG~1 Sheng WANG~1 Jian HOU~1 Yechun PEI~1 Ling LIU~1 Qing LI~1 Yongfu CHEN~1 Rongliang HU~2 Xiaorong AN~(1*) 1 State Key Laboratory of Agrobiotechnology Department of Biological sciences China Agricultural University Beijing 100094 China 2 The Insititue of Veterinary Medicine academy of military medical science of pla Changchun 130062 China

＜正＞The objective of this study was to coculture canine oocytes with canine oviductal cells or canine oviduct extracts in modified TCM199 culture medium to improve the MII *** canine oocytes were retrived from the l... 详细信息

关键词： canine oocytes oviductal cells 2-cell 8-cell parthenogenerated

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织物耐久抗菌整理的研究

染整技术 2003年第5期25卷 6-8,13页

作者：商成杰邹承淑张洪杰北京洁尔爽高科技有限公司北京100083 青岛大学医学院山东青岛266000 中国人民解放军军事医学科学院北京100000

介绍国外上世纪八、九十年代抗菌整理的动向和国内最近二十年来已在生产中应用的抗菌整理剂组成、整理方法和抗菌效果,对国外资料中的较成熟的抗菌性化合物也作了介绍。

关键词：织物耐久抗菌整理抗菌整理剂抗菌效果有机硅季铵盐二苯醚抗菌整理剂硝基呋喃类

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Development of Cell Lines Stably Expressing Staphylococcal Nuclease Fused to Dengue 2 Virus Capsid Protein for CTVI

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 577-582页

作者： Cheng-Feng QIN and E-De QIN Institute of Microbiology and Epidemiology, academy of military medical sciences, Beijing 100071, China Institute of Microbiology and Epidemiology Academy of Military Medical SciencesBeijing 100071 China

To explore the potential application of capsid-targeted viral inactivation (CTVI) strategy in prophylactic model against dengue virus (DV) infection, here we fused a Ca2+-dependent nuclease, staphylococcal nuclease (SN), to the capsid protein of dengue 2 virus (D2C) at the carboxyl terminal, and constructed the desired expression plasmid pc/D2C-SN and control plasmids pc/D2C-SN* and pc/D2C. A mammalian cell line BHK-21 was transfected by electroporation with those plasmids and thereafter selected by 5 μg/ml blasticidin. The resistant cell clones were then expanding cultured and screened by RT-PCR and Western Blot assays. The nuclease activity of the expressed fusion protein D2C-SN was analyzed by in vitro DNA digestion assay. It was confirmed cell lines stably expressing D2C-SN and control constructs were obtained. The intracellular expressed fusion protein D2C-SN had ideal nuclease activity and no cytotoxicity on mammalian cells. Those engineered cell lines provided the experimental system for CTVI application in prophylactic model and paved the new road for combating DV infection with CTVI.

关键词： dengue 2 virus capsid protein staphylococcal nuclease CTVI stable expression

Production of human liver prolidase by Saccharomyces cerevisiae as host cells

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Acta Pharmacologica Sinica 2004年第6期25卷 92-98页

作者： Shu-hao WANG, Min LIU, Mu-gen CHI, Qing-ding WANG, Man-ji SUN2 Institute of Pharmacologyand Toxicology, academy of military medical science, Beijing 100850, China Institute of Pharmacology and Toxicology Academy of Military Medical Science 北京 100850

AIM: To clone and express the recombinant human liver prolidase in yeast and explore the activities of both dipeptidaseand organophosphoric acid anhydrolase(OPAA). METHODS:The cDNAencodinghuman liver prolidase derived from healthy adult liver was cloned into the pYES2, an expression vector of S cerevisiae, and then trans- formed into S cerevisiae INVSc1 by electroporation. The transformant with the highest enzymatic activity was induced by galactose for expression. The optimal induction conditions (temperature, induction time, and the initial amount of inoculation cells) were estimated by orthogonal experimental design. The recombinant prolidase and OPAA activities were assayed by spectrocolorimetric methods. RESULTS: The recombinant enzyme catalyzed the hydrolysis of organophosphorous compound soman as well as the hydrolysis of dipeptide Gly-Pro. Under the optimal induction conditions (20 h, 25 oC, initial OD600=0.4), the maximum activities of prolidase and OPAA came to 226.5 and 578 μmol·min-1·g-1 protein in cell lysate supernatants, respectively. SDS-PAGE of the recombinant enzyme in disrupted cell supernatants showed a molecular weight of 56 kDa. Intensity scanning of the SDS-PAGE gel revealed that the enzyme accounted for 3.16 % of the total protein in the supernatant. One liter incubation medium produced 7 g of wet yeast cell containing 4.56 mg of the recombination protein. CONCLUSION: The recombinant human liver prolidase produced by yeast cell (S cerevisiae) exhibited both dipeptidase and OPAA activities.

关键词： prolidase liver human S Cerevisiae organophosphorous compounds

The Effect of CD3-Specific Monoclonal Antibody on Treating Experimental Autoimmune Myasthenia Gravis

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Cellular & Molecular Immunology 2005年第6期2卷 461-465页

作者： Ruonan Xu~1 Jianan Wang~1 Guojiang Chen~1 Gencheng Han~1 Renxi Wang~1 Beifen Shen~1 Yan Li~(1,2) ~1Institute of Basic medical science,academy of military medical science,Beijing 100850,China ~2Department of Molecular Immunology,Institute of Basic medical science,27 Taiping Road,Beijing 100850,China. Institute of Basic medical science Academy of Military Medical Science Beijing 100850 China

CD3-specific monoclonal antibody was the first one used for clinical practice in field of transplantation. Recently, renewed interests have elicited in its capacity to prevent autoimmune diabetes by inducing immune tolerance. In this study, we tested whether this antibody can also be used to treat another kind of autoimmune disease myasthenia gravis （MG） and explored the possible mechanisms. MG is caused by an autoimmune damage mediated by antibody- and complement-mediated destruction of AChR at the neuromuscular junction. We found that administration of CD3-specific antibody （Fab）2 to an animal model with experimental autoimmune myasthenia gravis （EAMG）（B6 mice received 3 times of AChR/CFA immunization） could not significantly improve the clinical signs and clinical score. When the possible mechanisms were tested, we found that CD3 antibody treatment slightly down-regulated the T-cell response to AChR, modestly up-regulation the muscle strength. And no significant difference in the titers of IgG2b was found between CD3 antibody treated and control groups. These data indicated that CD3-specific antibody was not suitable for treating MG, an antibody- and complementmediated autoimmune disease, after this disease has been established. The role of CD3-specific antibody in treating this kind of disease remains to be determined. Cellular ＆ Molecular Immunology. 2005;2（6）：461-465.

关键词： CD3-specific monoclonal antibody immune tolerance EAMG

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Cytotoxic T lymphocyte response induced by an improved synthetic lipopeptide vaccine against cervical cancer

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Acta Pharmacologica Sinica 2007年第5期28卷 695-702页

作者： Dao-hua XU~(1,5),Chen-hui ZHOU~2,Yong-peng XIA~3,Zong-ying QIU~3,Yu-zhang WU~4,Zheng-cai JIA~4,Wei ZHOU~4~1 Department of Pharmacology and ~2 School of Nursing,Guangdong medical College,Zhanjiang 524023,China ~3 Department of PharmacyChongqing University of medical science,Chongqing 400016,China,~4 Institute of Immunology pla,Third military medical University,Chongqing 400038,China Department of Pharmacology Guangdong Medical College Zhanjiang China School of Nursing Guangdong Medical College Zhanjiang China Department of Pharmacy Chongqing University of Medical Science Chongqing China Institute of Immunology PLA Third Military Medical University Chongqing China

Aim:To explore cytotoxic T lymphocyte （CTL） response induced by the lipopeptidevaccine against cervical ***:The immunological effect inducingCDS+ T cell-mediated cytotoxicity was investigated in human leukocyte antigen（HLA）-A2 transgenic mice and peripheral blood mononuclear cells （PBMC） ofhealthy HLA-A2.1+blood *** activity of specific CTL was measured byusing a standard 4 h51Cr release *** content of major histocompatibilitycomplex （MHC） I on T2 cells and the expression of immune molecules on dendriticcells （DC） were detected by flow cytometry,and the concentrations of interleukin（IL）-12 and interferon-γ were determined by ***:The lipopeptideinduced a strong epitope-specific CTL response both in vivo （transgenic mice）and in vitro （human PBMC）.This CTL induction was critically dependent on thepresence of the helper T lymphocyte epitope in transgenic mice,and the presenceof a lipid tail bypassed the need for an *** stability and persistence ofthe antigenic complex formed with the lipopeptide increased in comparison withthe CTL parental *** lipopeptide could induce the production of IL-12 inDC,but not the maturation of DC ***:The combination of CTLand the T helper epitope and lipid molecule can remarkably improve the immuno-genicity of the CTL peptide,the mechanism of which is associated with an in-crease in the stability and persistence of the antigenic complex formed with thelipopeptide and in the production of IL-12 in DC induced by the *** can be considered a more effective vaccine type for human being.

关键词： epitope cervical cancer lipopeptide vaccine

Construction and characterization of an experimental ISCOMS-based hepatitis B polypeptide vaccine

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World Journal of Gastroenterology 2002年第2期8卷 294-297页

作者： Xiao-Ju Guan Shanghai Institute of Materia Medica,Chinese academy of sciences,Shanghai 200031,China,previously worked as a postdoc in Institute of Immunology,Third military medical University,Chongqing 400038,China Xiao-Jun Guan,Department of science & Research,Second military medical University,Shanghai 200433,China Yu-Zhang Wu Zheng-Cai Jia Tong-Dong Shi Yan Tan,Institute of Immunology,Third military medical University,Chongqing 400038,China Shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai 200031 China.

AIM: To characterize the biochemical and immunological properties of an experimental ISCOMS vaccine prepared from a novel therapeutic polypeptide based on T cell epitopes of HBsAg, and a heptatis B-ISCOMS was prepared and investigated. METHODS: An immunostimulating complexes(ISCOMS)-based vaccine containing a novel therapeutic hepatitis B polypeptide was prepared by dialysis method, and its formation was visualized by electron microscopy and biochemically verified by SDS-polyacrylamide gel electrophoresis. Amount of the peptide within ISCOMS was determined by Bradford assay, and specific CTL response was detected by ELISPOT assay. RESULTS: Typical cage-like structures of submicroparticle with a diameter of about 40nm were observed by electron microscopy. Results from Bradford assay showed that the level of peptide incorporation was about 0.33g.L(-1). At the paralleled position close to the sixth band of the molecular weight marker(3480kDa) a clear band was shown in SDS-PAGE analysis, indicating successful incorporation of polypeptide into ISCOMS. It is suggested that ISCOMS delivery system could efficiently improve the immunogenicity of polypeptide and elicit specific immune responses in vivo by the results of ELISPOT assay, which showed that IFN-gamma producing cells(specific CTL responses) were increased(spots of ISCOMS-treated group: 47+/-5, n =3; control group: 5+/-2, n =3). CONCLUSION: ISCOMS-based hepatitis B polypeptide vaccine is successfully constructed and it induces a higher CTL response compared with short polypeptides vaccine in vivo.

关键词： Hepatitis B Vaccines ISCOMs Animals Enzyme-Linked Immunosorbent Assay Epitopes Female Hepatitis B Surface Antigens Humans Interferon Type II Mice Mice, Inbred BALB C Peptides Research Support, Non-U.S. Gov't T-Lymphocytes, Cytotoxic

Peptidergic innervation of human esophageal and cardiac carcinoma

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World Journal of Gastroenterology 2003年第3期9卷 399-403页

作者： Shuang-Hong Lü Yan Zhou Hai-Ping Que Shao-Jun Liu Department of Neurobiology, Institute of Basic medical sciences, academy of military medical sciences, Beijing 100850, China Department of Neurobiology Institute of Basic Medical Sciences Academy of Military Medical Sciences Beijing 100850 China.

AIM: To investigate the distribution of neuropeptideimmunoreactive nerve fibers in esophageal and cardiac carcinoma as well as their relationship with tumor cells so as to explore if there is nerve innervation in esophageal and cardiac ***: Esophageal and cardiac carcinoma specimens were collected from surgical operation. One part of them were fixed immediately with 4 % paraformaldehyde and then cut with a cryostat into 40-pm-thick sections to perform immunohistochemical analysis. Antibodies of ten kinds of neuropeptide including calcitonin gene-related peptide (CGRP), galanin (GAL), substance P (SP), etc. were used for immunostaining of nerve fibers. The other part of the tumor specimens were cut into little blocks (1 mm3) and cocultured with chick embryo dorsal root ganglia (DRG) to investigate if the tumor blocks could induce the neurons of DRG to extend processes, so as to probe into the possiblereasons for the nerve fibers growing into tumors. RESULTS: Substantial amounts of neuropeptide including GAL-, NPY-, SP-immunoreactive nerve bundles and scattered nerve fibers were distributed in esophageal and cardiac carcinomas. The scattered nerve fibers waved their way among tumor cells and contacted with tumor cells closely. Some of them even encircled tumor cells. There were many varicosities aligned on the nerve fibers like beads. They were also closely related to tumor cells. In the co-culture group, about 63 %and 67 % of DRG co-cultured with esophageal and cardiac tumor blocks respectively extended enormous processes,especially on the side adjacent to the tumor, whereas in the control group (without tumor blocks), no processes grew ***: Esophageal and cardiac carcinomas may be innervated by peptidergic nerve fibers, and they can induce neurons of DRG to extend processes in vitro.

关键词：肽能神经分布食管癌贲门癌肿瘤基因相关肽肿瘤生物学