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Imbalance of matrix metalloproteinase-9 and matrix metalloproteinase tissue inhibitor-1 may contribute to hemorrhage in cerebellar arteriovenous malformations

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Neural Regeneration Research 2012年第19期7卷 1513-1519页

作者： Fei Di Tongyan Chen Hongli Li Jizong Zhao Shuo Wang Yuanli Zhao Dong Zhang Department of Neurosurgery Beijing Tiantan Hospital Capital Medical University Beijing 100050 China Department of Laboratory Medicine Beijing Tiantan Hospital Capital Medical University Beijing 100050 China Department of Pathology Beijing Tiantan Hospital Capital Medical University Beijing 100050 China

In this study, we determined the expression levels of matrix metalloproteinase-2 and -9 and matrix metalloproteinase tissue inhibitor-1 and -2 in brain tissues and blood plasma of patients undergoing surgery for cerebellar arteriovenous malformations or primary epilepsy （control group）. Immunohistochemistry and enzyme-linked immunosorbent assay revealed that the expression of matrix metalloproteinase-9 and matrix metalloproteinase tissue inhibitor-1 was significantly higher in patients with cerebellar arteriovenous malformations than in patients with primary epilepsy. The ratio of matrix metalloproteinase-9 to matrix metalloproteinase tissue inhibitor-1 was significantly higher in patients with hemorrhagic cerebellar arteriovenous malformations compared with those with non-hemorrhagic malformations. Matrix metalloproteinase-2 and matrix metalloproteinase tissue inhibitor-2 levels were not significantly changed. These findings indicate that an imbalance of matrix metalloproteinase-9 and matrix metalloproteinase tissue inhibitor-I, resulting in a relative overabundance of matrix metalloproteinase-9, might be the underlying mechanism of hemorrhage of cerebellar arteriovenous malformations.

关键词： cerebellar arteriovenous malformations hemorrhage matrix metalloproteinase-2 matrixmetalloproteinase-9 tissue matrix metalloproteinase inhibitor-1 tissue matrix metalloproteinaseinhibitor-2 immunohistochemistry enzyme-linked immunosorbent assay neural regeneration

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Efficacy of catgut embedding in Baihui(GV20)and Feishu(BL13)and Pishu(BL20)on lung tissue,brain tissue and blood related indexes in rats with allergic rhinitis of lung deficiency type

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Journal of Traditional Chinese Medicine 2023年第5期43卷 973-982页

作者： HUANG Heyin WANG Yichen ZHANG Qinxiu WU Xiaojuan WANG Jiang Li Xiangyu PANG Rui FAN Li CAO Liu TANG Luo Department of ENT Sichuan Integrative Medicine HospitalChengdu 610041China Department of orthopaedics Sichuan Province Orthopaedic HospitalChengdu 610000China Department of ENT Affiliated Hospital of Chengdu University of Traditional Chinese MedicineChengdu 610000China Department of ENT Quxian Traditional Chinese Medicine HospitalQuxian 635299China

OBJECTIVE:To investigate the effects of acupoint catgut embedding for 3 weeks on lung tissue,blood immunoglobulin E(IgE)and interleutin-4(IL-4),brain tissue microglia x-42(OX-42)and toll-like receptor-2(TLR-2)in rats with allergic rhinitis of lung deficiency ***:Forty-five female Sprague-Dawley rats were randomly divided for two *** first time,they were randomly divided into model group and blank group(Group C)according to 2∶1,and the second time,the model group were randomly divided into model control group(Group B)and intervention treatment group(Group A)according to 1∶1.15 in each *** Group A and Group B,the lung deficiency model was made by"sulfur–moxa fumigation",and then the allergic rhinitis model was established by"ovalbumin(OVA)sensitization".Then catgut embedding was performed at acupoints in Group A and not in Group *** 3 weeks,collect lung tissue samples for hematoxylin-eosin staining,then take blood to observe the concentration of IgE and IL-4,and finally take brain tissue to observe the results of OX-42 and ***:IgE level(μg/m L)was(3.11±0.20)in the Group A,(4.19±0.44)in the Group B,and(2.29±0.30)in the Group C(all P<0.001).IL-4 level(pg/mL)was(14.2±0.7)in the Group A,(18.6±2.4)in the Group B,and(11.4±1.2)for the Group C(all P<0.001).The mean OD for OX-42 is(0.1728±0.0016)in the Group A,(0.1810±0.0046)in the Group B and(0.1674±0.0025)in the Group C(all P<0.001).CONCLUSION:Although 3 weeks of acupoint catgut embedding already showed obvious efficacy on rats with allergic rhinitis,the allergic reaction in the body still *** achieve further treatment,prolonging the catgut embedding time is necessary.

关键词： rhinitis,allergic acupoint catgut embedding enzyme-linked immunosorbent assay immunoglobulin E interleukin-4 hippocampus toll-like receptor 2

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Humoral and cellular immunogenecity of DNA vaccine based on hepatitis B core gene in rhesus monkeys

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World Journal of Gastroenterology 2001年第1期7卷 102-106页

作者： Zu Hu Huang1 Hui Zhuang2 Shan Lu3 Ren Hua Guo1 Guo Min Xu2 Jie Cai1 Wan Fu Zhu2 1Department of Infectious Diseases. The First Affiliated Hospital of Nanjing Medical University, Nenjing 210029, Jiangsu Province. China2Faculty of Microbiology, Beijing University, Beijing 100000, China3University of Massachusetts Medical Center The First Affiliated Hospital of Nanjing Medical University 300 Guangzhou Road Nanjing 210029 Jiangsu Province China.

INTRODUCTIONHepatitis B virus (HBV) is the most commonetiologic agent for infectious liver diseases. It isestimated that there are more than 250 millionchronic HBV carriersin the world today and thereis a significant ... 详细信息

关键词： Vaccines, DNA Animals Antibodies, Viral Antibody Formation Antibody Specificity Cell Division Cells, Cultured enzyme-linked immunosorbent assay Female Hepatitis B control Hepatitis B Core Antigens Immunity, Cellular Immunoglobulin G Interferon Type II Interleukin-4 Leukocytes, Mononuclear Macaca mulatta Male Research Support, Non-U.S. Gov't

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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

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Detection of anti-Helicobacter pyloriantibodies in serum and duodenal fluid in peptic gastroduodenal disease

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World Journal of Gastroenterology 2004年第20期10卷 2997-3000页

作者： Angelo Locateili Wilson Roberto Catapani Claudio Rufino Gomes Junior Claudilene Battistin Paula Silva Jaques Waisberg Department of Surgery ABC Faculty of MedicineAvenida Principe de Gales 821Santo AndreSao Paulo09060-650Brazil

AIM:To study the diagnosis of Helicobacter pylori(H pylori) infection through the determination of serum levels of anti- H pylori IgG and IgA antibodies,and the levels of anti-H pylori IgA antibodies in duodenal fluid. METHODS:Data were collected from 93 patients submitted to upper digestive endoscopy due to dyspeptic symptoms. The patients were either negative(group A)or positive (group B)to H pylori by means of both histological detection and urease *** endoscopy,peripheral blood was collected for the investigation of anti-H pylori IgG and IgA *** perform the urease test,biopsies were obtained from the gastric *** the histological evaluation,biopsies were collected from the gastric antrum (greater and lesser curvatures)and the gastric body. Following this,duodenal fluid was collected from the first and second portions of the *** the serological assaying of anti-Hpylori IgG and IgA,and anti-Hpylori IgA in duodenal fluids,the ELISA method was utilized. RESULTS:The concentration of serum IgG showed sensitivity of 64.0%,specificity of 83.7%,positive predictive value of 82.0%,negative predictive value of 66.6% and accuracy of 73.1% for the diagnosis of H pylori *** the same purpose,serum IgA showed sensitivity of 72.0%, specificity of 65.9%,positive predictive value of 72.0%, negative predictive value of 67.4% and accuracy of 69.8%. If the serological tests were considered together,*** both were positive or negative,the accuracy was 80.0%, sensitivity was 86.6%,specificity was 74.2%,positive predictive value was 74.2% and negative predictive value was 86.6%.When values obtained in the test for detecting IgA in the duodenal fluid were analyzed,no significant difference(P=0.43)was observed between the values obtained from patients with or without H pylori infection. CONCLUSION:The results of serum IgG and IgA tests for H pylori detection when used simultaneously,are more efficient in accuracy,sensitivity and negative pred

关键词： Adolescent Adult Aged Antibodies, Bacterial Duodenum Endoscopy, Gastrointestinal enzyme-linked immunosorbent assay Female Gastric Mucosa Helicobacter Infections Helicobacter pylori purification Humans Immunoglobulin A Immunoglobulin G Male Middle Aged Peptic Ulcer Sensitivity and Specificity Serologic Tests

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Preparation of single chain variable fragment of MG_7 mAb by phage display technology

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World Journal of Gastroenterology 2001年第4期7卷 510-514页

作者： Zhao-Cai Yu Jie Ding Yong-Zhan Nie Dai-Ming Fan Xue-Yong Zhang Department of Gastroenterology,Xijing Hospital,Fourth Military Medical University,Xi’an 710032,Shaanxi Province,China Department of Gastroenterology Xijing Hospital Fourth Military Medical University Xi'an 710032 Shaanxi Province China.

AIM: To develop the single chain variable fragment of MG MG(7)murine anti-human gastric cancer monoclonal antibody using the phage display technology for obtaining a tumor-targeting mediator. METHODS: mRNA was isolated from MG MG(7) producing murine hybridoma cell line and converted into cDNA. The variable fragments of heavy and light chain were amplified separately and assembled into ScFv with a specially constructed DNA linker by PCR. The ScFvs DNA was ligated into the phagmid vector pCANTAB5E and the ligated sample was transformed into competent E. Coli TG1. The transformed cells were infected with M13K07 helper phage to form MG MG(7) recombinant phage antibody library. The volume and recombinant rate of the library were evaluated by means of bacterial colony count and restriction analysis. After two rounds of panning with gastric cancer cell line KATO III of highly expressing MG(7)-binding antigen, the phage clones displaying ScFv of the antibody were selected by ELISA from the enriched phage clones. The antigen-binding affinity of the positive clone was detected by competition ELISA. HB2151 E. Coli was transfected with the positive phage clone demonstrated by competition ELISA for production of a soluble form of the MG(7) ScFv. ELISA assay was used to detect the antigen-binding affinity of the soluble MG(7) ScFv. Finally, the relative molecular mass of soluble MG(7) ScFv was measured by SDS-PAGE. RESULTS: The V(H), V(L) and ScFv DNAs were about 340bp, 320bp and 750bp, respectively. The volume of the library was up to 2 X 10(6) and 8 of 11 random clones were recombinants. Two phage clones could strongly compete with the original MG(7) antibody for binding to the antigen expressed on KATO III cells. Within 2 strong positive phage clones, the soluble MG(7) ScFv from one clone was found to have the binding activity with KATO III cells. SDS-PAGE showed that the relative molecular weight of soluble MG(7) ScFv was 32. CONCLUSION: The MG(7) ScFv was successfully produc

关键词： Peptide Library Animals Antibodies, Monoclonal Antibody Specificity Cell Line enzyme-linked immunosorbent assay Gene Therapy Genetic Screening Humans Hybridomas Immunoglobulin Variable Region Mice Recombination, Genetic Stomach Neoplasms

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Expression and identification of recombinant soluble single-chain variable fragment of monoclonal antibody MC3

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World Journal of Gastroenterology 2002年第2期8卷 258-262页

作者： Feng-Tian He Rong-Fen Li Yun-Sheng Kang Yan Zhang,Department of Biochemistry & Molecular Biology,Third Military Medical University,Chongqing 400038,China Yong-Zhan Nie Bao-Jun Chen Tai-Dong Qiao Dai-Ming Fan,Institute of Digestive Disease,Xijing Hospital,Fourth Military Medical University,Xi’an 710032,Shaanxi Province,China Department of Biochemistry & Molecular Biology Third Military Medical University Chongqing 400038 China. hefengtian@***

AIM: To generate soluble single chain variable fragments (ScFv) of monoclonal antibody MC3 recognizing colorectal and gastric carcinomas. METHODS: mRNA was isolated from the hybridoma cell line producing MC3 and the DNAs encoding variable domains of heavy and light chains (VH and VL) of the antibody were amplified separately by RT-PCR and assembled into ScFv DNA with a linker DNA. The ScFv DNA was ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into *** *** transformed cells were infected with M13KO7 helper phage to yield recombinant phages. After two rounds of panning with gastric carcinoma cell line AGS highly expressing MC3-binding antigen, the phage clones displaying ScFv fragments of the antibody were selected by ELISA. 4 phage clones showing strong signal in ELISA were used to infect *** HB2151 to express soluble ScFvs. The soluble ScFvs were identified by Dot blot and Western blot, and their antigen-binding activity was assayed by ELISA. The VH and VL DNAs of the ScFv DNA derived from phage clone 19 were sequenced. RESULTS: The VH,VL and ScFv DNAs were about 340 bp, 320 bp and 750 bp respectively. After two rounds of panning to the recombinant phages, 18 antigen-positive phage clones were selected from 30 preselected phage clones by ELISA. All the soluble ScFvs derived from the 4 out of the 18 antigen-positive phage clones were about M(r)32000 and concentrated in periplasmatic space under the given culture condition. The soluble ScFvs could bind the antigen, and they shared the same binding site with MC3. The sequences of the VH and VL DNAs of the MC3 ScFv showed that the variable antibody genes belonged to the IgG1 subgroup,kappa-type. CONCLUSION: The soluble ScFv of MC3 is successfully produced, which not only provides a possible novel targeting vehicle for in vivo and in vitro study on associated cancers, but also offers the antibody a stable genetic source.

关键词： Animals Antibodies, Monoclonal Base Sequence Carcinoma Colorectal Neoplasms enzyme-linked immunosorbent assay Humans Immunoglobulin Fragments Immunoglobulin Variable Region Mice Molecular Sequence Data Recombinant Proteins Stomach Neoplasms Tumor Cells, Cultured

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Prokaryotical expression of structural and non-structural proteins of hepatitis G virus

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World Journal of Gastroenterology 2001年第5期7卷 642-646页

作者： Ning-Shao Xia~1 Hai-Jie Yang~1 Jun Zhang~1 Chang-Qing Lin~1 Ying-Bin Wang~1 Juan Wang~1 Mei-Yun Zhan~2 MH Ng~3 1 Key Laboratory of the Ministry of Education for Cell Biology and Tumor Cell Engineering,Xiamen University,Xiamen 361005,Fujian Province,China2 Institute of Virology,Chinese Academy of Preventive Medicine Beijing 100052,China3 Department of Microbiology,Hoog Kong University,Hongkong,China Key Laboratory of the Ministry of Education for Cell Biology and Tumor Cell Engineering Xiamen University Xiamen 361005 Fujian Province China. nsxia@***

AIM: To study the epitope distribution of hepatitis G virus (HGV) and to seek for the potential recombinant antigens for the development of HGV diagnostic reagents. METHODS: Fourteen clones encompassing HGV gene fragments from core to NS3 and NS5 were constructed using prokaryotic expression vector pRSET and (or) pGEX, and expressed in ***. Western blotting and ELISA were used to detect the immunoreactivity of these recombinant proteins. RESULTS: One clone with HGV fragment from core to E1 (G1), one from E2 (G31), three from NS3 (G6, G61, G7), one from NS5B (G821) and one chimeric fragment from NS3 and NS5B (G61-821) could be expressed well and showed obvious immunoreactivity by Western blotting. One clone with HGV framment from NS5B (G82) was also well expressed, but could not show immunoreactivity by Western blotting. No obvious expression was found in the other six clones. All the expressed recombinant proteins were in inclusion body form, except the protein G61 which could be expressed in soluble form. Further purified recombinant proteins G1, G31, G61, G821 and G61-821 were detected in indirected ELISA as coating antigen respectively. Only recombinant G1 could still show immunoreactivity, and the other four recombinant proteins failed to react to the HGV antibody positive sera. Western blotting results indicated that the immunoactivity of these four recombinant proteins were lost during purification. CONCLUSION: Core to E1, E2, NS3 and NS5 fragment of HGV contain antigenic epitopes, which could be produced in prokaryotically expressed recombinant proteins. A high-yield recombinant protein (G1) located in HGV core to E1 could remain its epitope after purification, which showed the potential that G1 could be used as a coating antigen to develop an ELISA kit for HGV specific antibody diagnosis.

关键词： Blotting, Western enzyme-linked immunosorbent assay Epitope Mapping Escherichia coli GB virus C purification Gene Expression Regulation, Viral Humans Plasmids Recombinant Proteins Viral Envelope Proteins Viral Nonstructural Proteins

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Interaction of hepatitis C virus envelope glycoprotein E2 with the large extracellular loop of tupaia CD81

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World Journal of Gastroenterology 2009年第2期15卷 240-244页

作者： Zhan-Fei Tian Hong Shen Xi-Hua Fu Yi-Chun Chen Hubert E Blum Thomas F Baumert Xi-Ping Zhao Department of Infectious Diseases 161 Hospital of PLA Wuhan 430030 Hubei Province China Department of Infectious Diseases Tongji Hospital Tongji Medical College Huazhong Science & Technology University Wuhan 430030 Hubei Province China Department of Medicine II University of Freiburg D-79106 Freiburg Germany Inserm U748 Universit6 Louis Pasteur Service d'Hepatogastroenterologie 3 Rue Koeberle F-67000 Strasbourg France

AIM: To further analyze the interaction of tupaia CD81 with hepatitis C virus (HCV) envelope protein E2. METHODS: A tupaia CD81 large extracellular loop (CD81 LEL), which binds to HCV E2 protein, was cloned and expressed as a GST-fusion protein, and interaction of HCV E2 protein with a tupaia CD81 LEL was evaluated by enzyme-linked immunosorbent assay (EIA). RESULTS: Although tupaia and human CD81 LEL differed in 6 amino acid changes, tupaia CD81 LEL was strongly recognized by anti-CD81 antibodies against human CD81 LEL conformation-dependent epitopes. Investigating LEL CD81-E2 interactions by EIA, we demonstrated that binding of tupaia CD81 LEL GST fusion protein to recombinant HCV E2 protein was markedly reduced compared to binding of human CD81 LEL GST fusion protein to recombinant HCV E2 protein. CONCLUSION: These data suggest that the structural differences in-between the tupaia and human CD81 may alter the interaction of the large extracellular loop with HCV envelope glycoprotein E2. These findings may be important for the understanding of the mechanisms of binding and entry of HCV to PTHs.

关键词： Hepatitis C virus E2 protein Tupaia CD81,Bind enzyme-linked immunosorbent assay

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Levels of v5 and v6 CD44 splice variants in serum of patients with colorectal cancer are not correlated with pT stage,histopathological grade of malignancy and clinical features

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World Journal of Gastroenterology 2004年第4期10卷 583-585页

作者： Bogdan Zalewski 2nd Department of General and Gastroenterological Surgery Medical University of BialystokPoland

AIM:This study was designed to compare the levels of v5 and v6 splice variants of CD44 evaluated using EITSA test in the serum of patients with colorectal cancer in different stages of progression of the disease estimated in pT stage according to WHO score,histopathological grade of malignancy and some clinicopathological features. METHODS:The serum obtained from 114 persons with colorectal adenocarcinomas was examined using ELISA method,pT stage and grade of malignancy of the tumour were examined in formalin fixed and paraffin embedded materials obtained during operation. RESULTS:Only the level of CD44 v5 in the serum of patients before operation with G2 pT4 tumour was lower than that in other probes and the difference was statistically significant. We did not find any other correlations between the level of v5 and v6 CD44 variants and other evaluated parameters. CONCLUSION:The level of CD44 v5 and v6 estimated by ELISA test in the serum can not be used as a prognostic factor in colorectal cancer.

关键词： Alternative Splicing Adenocarcinoma Adult Aged Aged, 80 and over Antigens, CD44 Colorectal Neoplasms Disease Progression enzyme-linked immunosorbent assay Female Humans Male Middle Aged Predictive Value of Tests Prognosis Tumor Markers, Biological

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