CLINICAL SIGNIFICANCE OF DETECTING SERUM ANTIBODIES TO NUCLEOPROTEIN AND GLYCOPROTEIN G_2 OF HANTAAN VIRUS IN PATIENTS WITH HEMORRHAGIC FEVER WITH RENAL SYNDROME

Hantaanvirus,theetiologicagentofhemorrhagicfeverwithrenalsyndrome(HFRS),canleadtoaseveretypeofHFRSandbelongstothegenusHantavirusofthefamilyBunyaviridae.ThegenomeofHantannvirusconsistsofthreesegmentsL,MandSrespectivelyencodingpolymerase,membraneglycoproteinG,andG,andnucleoproteinofthevirus.Inthepresentpaper,serumantibodiestonecleoprotein(NPAb)andglycoproteinG,(G,Ab)ofHantaanvirusinpatientswithHFRSweredynamicallydeterminedusingantibodyblockingenzymelinkedimmunosorbentassays(B--ELISA…     

新型杂多化合物抗禽流感病毒的实验研究

目的探讨新型杂多化合物(PTW)抗禽流感病毒的作用。方法采用细胞病变法结合MTT法检测PTW对MDCK细胞的毒性和对禽流感病毒(H5N1)的抑制作用;以H5N1感染的小鼠为实验模型,检测PTW对H5N1感染小鼠的治疗作用。结果PTW对MDCK细胞的最大无毒质量浓度为190.55 mg/L,半数致死质量浓度为909.9 mg/L;体外实验结果表明PTW对禽流感病毒半数抑制质量浓度(IC50)为13.49 mg/L,治疗指数(TI)为67.44;体内实验显示PTW对H5N1感染小鼠有明显的治疗作用,25 mg/kg剂量组对H5N1感染小鼠的肺炎抑制率达166.16%,生命延长率为28.36%。结论PTW对H5N1禽流感病毒具有较好的抑制作用。     

构建由胸腺嘧啶激酶基因—小Mu噬菌体DNA插入到HSV BamHI C片段的重组质粒

为了使单纯疱疹病毒1型(HSV-1)的BamHI C片段发生突变,利用DNA重组技术将胸腺嘧啶激酶基因-小Mu噬菌体DNA系统(TK—mM)随机插入到质粒pRB 132中的HSV-1 BamHI C片段内,用DNA斑点杂交法得到6个在BamHI C片段上有TK-mM插入的重组质粒。限制性核酸内切酶的分析结果表明,由于TK—mM的插入,BamHI C片段发生突变,而TK—mM在6个重组质粒中的插入位点也是各不相同的。     

血清HBV标志物阳性和阴性肾炎患者的临床与病理分析

目的　探讨乙型肝炎病毒 (HBV)感染对肾小球肾炎 (GN)的致病作用。方法　应用免疫组织化学方法 ,检测 2 8例血清HBV阳性和 1 2例HBV阴性组患者肾组织内HBV抗原(HBAg)存在的状况 ,并着重对比了两组患者的临床表现和肾组织病变特征。结果　HBV标志物阳性组HBAg检出率较高 (71 .43 % ) ,肾小球内增生病变较重 ,但统计学处理两组间在临床表现和病变之间均无显著性差异。结论　HBV可能直接感染肾组织细胞导致肾炎的发生。     

THE INVESTIGATION OF DIFFERENT POPULATION＇S TRANSFUSION-TRANSMITTED VIRUS-DNA IN SHAANXI PROVINCE

Objective To investigate transfusion transmitted virus (TTV) infection among population of different groups in Shaanxi Province. Methods A nested polymerase chain reaction (PCR) with primers from ORF1 of TTV genome was established to detect TTV-DNA in serum of the patients. Results TTV-DAN was detected in the sera of 3 of 50 cases of general population(6％), 2 of 30 cases of vocational blood donors(6.7%),21 of 97 cases with Type B hepatitis (21. 6％), 9 of 35 cases of Type C hepatitis (25. 7％),and 23 of 40 cases with non-A～ non-G hepatitis (57.5 ％ ). Conclusion There is TTV infection among general population in Shaanxi Province. TTV may be an impor- tant agent to cause non-A～non-G hepatitis. And the patients with HBV or HCV can have overlapping TTV infection.     

Asymmetric PCR method in generation of HBV ssDNA for pyrosequencing

Objective To explore the optimal primer ratio and concentration of asymmetric polymerase chain reaction (A-PCR) in producing hepatitis B virus (HBV) single-stranded DNA (ssDNA) for pyrosequencing. Methods A-PCR was carried out to generate HBV ssDNA with forward to reverse primers of different ratios (50 : 1, 100 : 1) and concentrations (13. 0 pmol/25μL and 0.14 pmol/25μL, 19. 5 pmol/25μL and 0. 21 pmol/25μL), and the product yield and quality were compared respectively. Results The forward to reverse primer ratio of 50 : 1 provided better yield and concentration of 19. 5 pmol/25μL and 0. 21 pmol//25μL generated a clearer band. Conclusion A simple and feasible method to produce HBV ssDNA for pyrosequencing in batch is established.     

乙型肝炎病毒基因型检测及其与肝功能的关系

目的 探讨乙型肝炎病毒基因型与肝功能的关系.方法 采用微板核酸分子杂交ELISA法检测93例不同临床类型的乙型肝炎患者的基因型并同时测定肝功能.结果 93例不同临床类型乙型病毒性肝炎患者中,B型24例(25.81%),C型59例(63.44%),D型5例(5.38%),混合型5例(B/D 3例,C/D 2例,占5.38%).93例不同临床类型的乙型肝炎患者中,以C基因型为主,其次为B基因型,部分以D型和混合型存在,无A、E、F型.按照慢性乙型肝炎、亚急性重型肝炎、肝硬化的顺序,C基因型的检出率逐渐增多,B基因型的检出率逐渐减少.肝癌患者C基因型的检出率没有依次增高.C基因型各型乙型肝炎患者的ALT、AST、TBIL均高于B型,ALB低于B型,但差别无统计学意义.结论 西安地区病毒性肝炎以C基因型为主,部分以B、D及混合型存在,未发现A、E、F型.除肝癌外,C型的检出率随临床类型的加重而增多.     

Screening of hepatocyte proteins binding to NS5ABP37 protein by yeast-two hybrid system

Objective To investigate the biological function of NS5ABP37 and to look for proteins interacting with NS5ABP37 protein in hepatocytes. Methods We constructed bait plasmid expressing NS5ABP37 protein of hepatitis C virus (HCV) by cloning the gene of NS5ABP37 protein into pGBKT7, then the recombinant plasmid DNA was transformed into yeast AH109 (α type). The transformed yeast AH109 was mated with yeast Y187 (α type) containing liver cDNA library plasmid in 2×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-α-gal for selection and screening. After extracting and sequencing of plasmids from positive (blue) colonies, we made a sequence analysis by bioinformatics. Results We screened twenty-five proteins binding to NS5ABP37, including Homo sapiens cyclin Ⅰ (CCNI) gene, Homo sapiens matrix metallopeptidase 25 (MMP25) and Homo sapiens talin 1. Conclusion The yeast-two hybrid system is an effective method for identifying hepatocyte proteins interacting with NS5ABP37 of HCV. And the biological function of NS5ABP37 may be associated with glycometabolism, lipid metabolism and apoptosis.     

2型腺伴随病毒基因治疗载体的构建

目的　构建一种适用于中枢神经系统疾病的基因治疗载体。方法　以野生型 2型腺伴随病毒 (AAV 2 )质粒pssv9int 及逆转录病毒载体质粒plxsn为基本元件 ,利用重组DNA技术 ,构建了一种通用型AAV载体质粒pssHG Neo。结果　该质粒除含有必备的原核细胞复制子及Ampr 筛选基因外 ,还含有真核细胞筛选基因———Neor 及供插入目的基因的多克隆位点 (MCS) ,在MCS的上游为逆转录病毒的长末端重复序列 (5’LTR) ,具有启动子的功能 ,启动定向插入到MCS的外源目的基因。质粒中的AAV反向末端重复序列 (ITR)具有定点整合于人基因组 1 9号染色体S1位点的特点。结论　AAV载体是具有广阔前景的基因治疗载体     

流行性出血热患者尿中病毒抗原的观察

本文报道了采用ELISA双夹心法、ELISA阻断试验、沉淀试验、SPA协同凝集试验、补体结合试验检测流行出血热(EHF)患者尿中流行性出血热病毒(EHFV)抗原;并用免疫电镜观察了尿中流行性出血热病毒形态;用尿中流行性出血热病毒抗原免疫家兔观察了其免疫源性。实验表明:①用ELISA双夹心法检测EHF患者尿抗原,早期检出率高、重复性好、特异性强,较上述其它检测尿抗原的方法好。②上述多种方法均证明EHF患者尿中确有EHFV抗原排出。③该EHFV抗原还具有一定的免疫源性。提示:ELISA双夹心法适用于EHF临床诊断及流行病学调查。