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

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

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THE ROLE OF PERIAQUEDUCTAL GRAY (PAG) IN THE ACUPUNCTURE ANALGESIA AND MODULATION OF PAIN

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针刺研究 1989年第Z1期 87-88页

作者： Zhang Changcheng Li Xicheng Department of Physiology, The third military medical university, Chongqing

The role of PAG in the acupuncture analgesia 1. After noxious stimulation of splanchnic nerve or tail, the majority ofunits responded with increased frequency, some of units showed decreased fre-quency and the remainder had no response to both of noxious stimulation. Itseems to imply that PAG is one of the brain structures which receive somati-co-visceral pain impulses.

关键词： PAG IN THE ACUPUNCTURE ANALGESIA AND MODULATION OF PAIN THE ROLE OF PERIAQUEDUCTAL GRAY

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Stable Skin-specific Overexpression of Human CTLA4-Ig in Transgenic Mice through Seven Generations

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Acta Biochimica et Biophysica Sinica 2006年第3期38卷 171-178页

作者： Yong WANG~1 Yong NI~(1,3) Hong WEI~1* Feng-Chao WANG~2 Liang-Peng GE~1 Xiang GAO~41 Department of Animal Science,College of Basic Medicine,third military medical university,Chongqing 400038,China 2 Institute of Combined Injury,College of military Preventive Medicine,third military medical university,Chongqing 400038,China 3 School of Life Sciences,Southwest university,Chongqing 400715,China 4 Animal Model Research Institute,Nanjing university,Nanjing 210093,China Department of Animal Science College of Basic Medicine Third Military Medical UniversityChongqing 400038 China School of Life Sciences Southwest UniversityChongqing 400715 China Institute of Combined Injury College of Military Preventive Medicine Third Military Medical UniversityChongqing 400038 China Animal Model Research Institute Nanjing UniversityNanjing 210093 China

Skin graft rejection is a typical cellular immune response,mainly mediated by T *** lymphocyte associated antigen 4-immunoglobin (CTLA4-Ig) extends graft survival by blocking the T cellco-stimulation pathway and inhibiting T cell *** investigate the efficacy of CTLA4-Ig in prolong-ing skin graft survival,human CTLA4-Ig (hCTLA4-Ig) was engineered to overexpress in mouse skin bytransgenesis using the K14 *** transcription-polymerase chain reaction (RT-PCR) and West-ern blot assay indicated that the expression of CTLA4-Ig remained skin-specific and relatively constantcompared to the internal control protein,AKT,through seven *** presence and concentrationof the hCTLA4-Ig protein in transgenic mouse sera was determined by enzyme-linked immunosorbent assay(ELISA),and the results indicated that the serum CTLA4-Ig concentration also remained constant *** of transgenic mouse skins grafted onto rat wounds was remarkably prolonged com-pared to that of wild-type skins from the same mouse strain,and remained comparable among all *** suggested that the bioactive hCTLA4-Ig protein was stably expressed in transgenical micethrough at least seven generations,which was consistent with the stable skin-specific CTLA4-Ig *** results demonstrated that the transgenic expression of hCTLA4-Ig in skin driven by the K14 promoterremained constant through generations,and a transgenic line can be established to provide transgenic skinwith extended survival reproducibly.

关键词： cytotoxic T lymphocyte antigen 4-Ig transgenic mice skin K14 promoter

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Prenatal exposure to lipopolysaccharide results in increases in blood pressure and body weight in rats

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Acta Pharmacologica Sinica 2007年第5期28卷 651-656页

作者： Yan-ling WEI Xiao-hui LI Jian-zhi ZHOU Institute of Materia Medica,School of Basic Medicine,third military medical university,Chongqing 400038,China Institute of Materia Medica School of Basic Medicine Third Military Medical University Chongqing China

Aim:To investigate the effects of prenatal exposure to lipopolysaccharide (LPS)on blood pressure and body weight of offspring in ***:Sixteen healthy,pregnant rats were randomly divided into 2 *** rats in the LPS groupwere injected intraperitoneally with LPS (0.79 mg/kg) on the d 8,d 10,and d 12 *** in the control group were only treated with normal ***,all offspring were weighed and blood pressure was measured by the tail-cuff method once every 2 weeks from the 6th to the 24th *** the 15th week,their food intake was weighed every *** the end of the 24th week,the ratswere killed by *** adipose tissues were weighed,and theserum level of leptin was detected by ***:The offspringwith prenatal LPS exposure showed increased systemic arterial pressure,heavierbody weight,elevated food intake,increased adipose tissue weight,and increasedcirculating leptin compared with the ***:Prenatal exposure toLPS leads to increases in blood pressure and body weight in rats.

关键词： hypertension inflammation obesity prenatal exposure

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POTENTIATION EFFECT OF DIAZEPAM ON ACTH ANALGESIA IN THE MICE

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针刺研究 1989年第Z1期 117-118页

作者： Li Haiti Zhang Wende Department of Physiology, third military medical university, Chongqing

In the present study we injected diazepam, r-aminobutyric acid (GABA)receptor antagonist——picrotoxin and GABA synthesis inhibitor——isoniazid into the intraperitoneal (ip). Mice painful threshold and serotonin (5-HT)content in mice brain were measured by spectrofluorometric assay, the effectof diazepam on ACTH analgesia was obstraeted. The possible mechanisms ofdiazepam effect were studied.

关键词： ACTH POTENTIATION EFFECT OF DIAZEPAM ON ACTH ANALGESIA IN THE MICE

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Analysis of spontaneous, gamma ray-and ethylnitrosourea-induced hprt mutants in HL-60 cells with multiplex PCR

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World Journal of Gastroenterology 2003年第3期9卷 578-583页

作者： Sheng-Xue Liu Jia Cao Hui An Hua-Min Shun Lu-Jun Yang Yong Liu Department of Health Toxicology,Preventive medical College,third military medical university,Chongqing 400038,China Department of Healath Toxicology Preventive Medical College Third Military Medical University Chongqing 400038 China.

AIM: To explore the molecular spectra and mechanism of human hypoxanthine guanine phosphoribosyl transferase (hprt) gene mutation induced by ethyluitrosourea (ENU) and 60Co γ-***: Independent human promyelocytic leukemia cells (HL-60) mutants at the hprt locus were isolated from untreated, ethyluitrosourea (ENU) and 60Co γ-ray-exposed cells, respectively, and verified by two-way screening. The genetic changes underlying the mutation were determined by multiplex polymerase chain reaction (PCR) amplification and electrophoresis ***: With dosage increased, survival rate of plated cell reduced (in the group with dosage of ENU with 100-200μg/ml, P＜0.01; in the group with dosage of 60Co γ-ray with 2-4 Gy, P＜0.05) and mutational frequency increased (in the group of ENU 12.5-200.0 μg/ml, P＜0.05; in the group of 60Co γ-ray with 1-4 Gy, P＜0.05) significantly. In the 13spontaneous mutants analyzed, 92.3 % of mutant clones did not show any change in number or size of exon, a single exon was lost in 7.7 %, and no evidence indicated total gene deletion occurred in nine hprt exons. However, deletions were found in 79.7 % of ENU-induced mutations (62.5-89.4 %,P＜0.01) and in 61.7 % of gamma-ray-induced mutations (28.6-76.5 %, P＜0.01). There were deletion mutations in all 9 exons of hprt gene and the most of induced mutations were chain deletion with multiplex exons (97.9 % in gammaray-induced mutants, 88.1% in ENU-induced mutants).CONCLUSION: The spectra of spontaneous mutations differs completely from that induced by EUN or 60Co γ-*** both ENU and γ-ray can cause destruction of genetic structure, mechanism of mutagenesis between them may be different.

关键词：人次黄嘌呤鸟嘌呤转磷酸糖基酶 HL-60细胞 γ射线多重PCR 基因突变分子生物学

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STUDIES ON THE REACTIONS OF HIPPOCAMPAL FORMATION IN PAIN MODULATION AND ACUPUNCTURE ANALGESIA

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针刺研究 1989年第Z1期 45-46页

作者： Liu Zuozhou Liao Weihong Department of Physiology, The third military medical university, Chongqing, China

It has been proved that the hippocampal formation is closely related tosome behaviour reactions (such as learning and memory), visceraI activities,and neuro-endocrine regulations. However, what roles the hippocampus mayplay in pain modulation and analgesic mechanism still need further *** we have done some works in this field. The blood pressure increased accompanying hippocampal EEG's slowrhythmic activity (SRA or theta rhythm) which had a higher amplitude and alonger time course. This EEG wave was specific and could be used as one ofindexes of nociceptive response. Our experiments also demonstrated that theblood pressure increase could be induced by peripheral somatic noxious stimu-

关键词： EEG STUDIES ON THE REACTIONS OF HIPPOCAMPAL FORMATION IN PAIN MODULATION AND ACUPUNCTURE ANALGESIA

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Mechanical properties of hepatocellular carcinoma cells

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World Journal of Gastroenterology 2002年第2期8卷 243-246页

作者： Gang Zhang,Department of Pathophysiology,The third military medical university,Chongqing 400038,China Mian Long Zhe-Zhi Wu Wei-Qun Yu,College of Bioengineer,Chongqing university,Chongqing 400044,China Department of Pathophysiology The Third Military Medical University Chongqing 400038 China. a65412423@***

AIM: To study the viscoelastic properties of human hepatocytes and hepatocellular carcinoma (HCC) cells under cytoskeletal perturbation, and to further to study the viscoelastic properties and the adhesive properties of mouse hepatoma cells (HTC) in different cell cycle. METHODS: Micropipette aspiration technique was adopted to measure viscoelastic coefficients and adhesion force to collagen coated surface of the cells. Three kinds of cytoskeleton perturbing agents, colchicines (Col), cytochalasin D (CD) and vinblastine (VBL), were used to treat HCC cells and hepatocytes and the effects of these treatment on cell viscoelastic coefficients were investigated. The experimental results were analyzed with a three-element standard linear solid. Further, the viscoelastic properties of HTC cells and the adhesion force of different cycle HTC cells were also investigated. The synchronous G(1) and S phase cells were achieved through thymine-2-desoryriboside and colchicines sequential blockage method and thymine-2-desoryriboside blockage method respectively. RESULTS: The elastic coefficients, but not viscous coefficient of HCC cells (K(1)=103.6+/-12.6N.m(-2), K(2)=42.5 +/ 10.4N.m(-2), mu=4.5 +/- 1.9Pa.s), were significantly higher than the corresponding value for hepatocytes (K(1)=87.5 +/- 12.1N.m(-2), K(2)=33.3+/-10.3N.m(-2), mu=5.9+/-3.0Pa.s, P<0.01). Upon treatment with CD, the viscoelastic coefficients of both hepatocytes and HCC cells decreased consistently, with magnitudes for the decrease in elastic coefficients of HCC cells (K(1): 68.7 N.m(-2) to 81.7N.m(-2), 66.3% to 78.9%; K(2): 34.5N.m(-2) to 37.1N.m(-2), 81.2% to 87.3%, P<0.001) larger than those for normal hepatocytes (K(1): 42.6N.m(-2) to 49.8N.m(-2), 48.7% to 56.9%; K(2): 17.2N.m(-2) to 20.4N.m(-2), 51.7% to 61.3%, P<0.001). There was a little decrease in the viscous coefficient of HCC cells (2.0 to 3.4Pa.s, 44.4 to 75.6%, P<0.001) than that for hepatocytes (3.0 to 3.9Pa.s, 50.8 to 66.1% P<0.001).

关键词： Animals Antineoplastic Agents, Phytogenic Carcinoma, Hepatocellular Cell Adhesion Cell Cycle Colchicine Cytochalasin D Cytoskeleton Elasticity Hepatocytes Humans Liver Neoplasms Mice Nucleic Acid Synthesis Inhibitors Research Support, Non-U.S. Gov't Tumor Cells, Cultured Vinblastine

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Distribution of nitric oxide synthase in stomach myenteric plexus of rats

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World Journal of Gastroenterology 2001年第6期7卷 852-854页

作者： Xi Peng Jin-Bin Feng Hong Yan Yun Zhao Shi-Liang Wang Institute of Burn Research,Southwest Hospital,third military medical university,Chongqing 400038,China Institute of Burn Research Southwestern Hospital Third Military Medical University Chongqing 400038 China. pxlr@***

AIM: To study the distribution of nitric oxide synthase (NOS) in rat stomach myenteric plexus. METHODS: The distribution of NOS in gastric wall was studied in quantity and location by the NADPH-diaphorase (NDP) histochemical staining method and whole mount preparation technique. RESULTS: NOS was distributed in whole stomach wall, most of them were located in myenteric plexus, and distributed in submucosal *** shape of NOS positive neurons was basically similar, most of them being round and oval in shape. But their density, size and staining intensity varied greatly in the different parts of stomach. The density was 62+/-38 cells mm(2) (antrum), 43+/-32 cells/mm(2) (body), and 32+/-28 cells mm(2) (fundus), respectively. The size and staining intensity of NOS positive neurons in the fundus were basically the same, the neurons being large and dark stained, while they were obviously different in antrum. In the body of the stomach, the NOS positive neurons were in an intermediate state from fundus to antrum. There were some beadlike structures which were strung together by NOS positive varicosities in nerve fibers, some were closely adherent to the outer walls of blood vessels. CONCLUSION: Nitric oxide might be involved in the modulation of motility, secretion and blood circulation of the stomach, and the significant difference of NOS positive neurons in different parts of stomach myenteric plexus may be related to the physiologic function of stomach.

关键词： Animals Female Myenteric Plexus Nitric Oxide Synthase Rats Rats, Wistar Stomach Tissue Distribution

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Diagnosis and treatment of portal biliopathy

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Hepatobiliary & Pancreatic Diseases International 2002年第4期1卷 581-586页

作者： Zhen-Ping He Lin-Jun Fan the Southwest Hepatobiliary Surgery Institute, third military medical university, Chongquing 400038, China Southwest Hepatobiliary Surg. Inst. Third Military Medical University Chongqing 400038 China

Background: Portal biliopathy producing intrahepa- tic and extrahepatic biliary ductai abnormalities from portal hypertension, particularly with extrahepatic portal vein obstruction (EHPVO) is common. A majority of these patients are asymptomatic, but oc- casionally there is symptomatic biliary obstruction, and cholangitis and choledocholithiasis. Objective: To explore the principles of diagnosis and treatment of portal biliopathy. Data sources: To review the literature of portal bili- opathy. Conclusions: Endoscopic sphincterotomy, stone ex- traction and supportive drainage could effectively re- lieve cholangitis when jaundice is associated with common bile duct stones. Definitive decompressive portal-system vein shunting operation and choledo- cho-jejunostomy are sometimes required when biliary obstruction is recurrent and progressive.

关键词： portal biliopathy extrahepatic portal vein obstruction diagnosis treatment shunt

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