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大肠杆菌表达的戊型肝炎病毒ORF2片段的聚合现象研究 被引量:37
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作者 李少伟 张军 +5 位作者 何志强 葛胜祥 顾颖 林鉴 刘如石 夏宁邵 《生物工程学报》 CAS CSCD 北大核心 2002年第4期463-467,共5页
在大肠杆菌中表达了戊型肝炎病毒 (HEV)ORF2的a .a .394~a .a .6 0 4片段 ,得到的重组蛋白NE2在SDS PAGE中主要以可被尿素解聚的二聚体形式存在 ,二聚体对病人血清的反应性明显强于单体 ;质谱分析表明NE2可形成从二聚体到至少六聚体的... 在大肠杆菌中表达了戊型肝炎病毒 (HEV)ORF2的a .a .394~a .a .6 0 4片段 ,得到的重组蛋白NE2在SDS PAGE中主要以可被尿素解聚的二聚体形式存在 ,二聚体对病人血清的反应性明显强于单体 ;质谱分析表明NE2可形成从二聚体到至少六聚体的多种聚体 ;动态光散射测定表明平均分子半径约 4nm ,相当于四聚体 ,但分散度较大 ,提示为多种大小不一的聚合体的混合物。这些证据表明NE2蛋白可形成以同源二聚体为基本单位的多种聚合体形式 ,其中以二聚体间的结合最为紧密 ,并且以二聚体为基础可进一步装配出多种更高级结构 ,从而具有作为HEV疫苗及诊断试剂抗原的良好前景。 展开更多
关键词 大肠杆菌 表达 肝炎病毒ORF2片段 聚合现象 组装 二聚体 原核表达
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2a/Ⅲ型丙型肝炎病毒基因组非结构4区全长序列的扩增、克隆及分析
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作者 陈勇 封波 +1 位作者 丛旭 魏来 《徐州医学院学报》 CAS 2002年第2期95-99,共5页
目的 获得 2a/Ⅲ型丙型肝炎病毒 (HCV)基因组全长非结构 (NS) 4区克隆并作序列分析。方法 用限制性片段长度多态性分析 (RFLP)基因分型方法筛选出 2a/Ⅲ型HCV一株。为了获得该毒株全长NS4区基因序列 ,依据代表株序列的NS3区之 3′末端... 目的 获得 2a/Ⅲ型丙型肝炎病毒 (HCV)基因组全长非结构 (NS) 4区克隆并作序列分析。方法 用限制性片段长度多态性分析 (RFLP)基因分型方法筛选出 2a/Ⅲ型HCV一株。为了获得该毒株全长NS4区基因序列 ,依据代表株序列的NS3区之 3′末端及NS5区之 5′末端相对保守区域合成引物 ,以RT -nested -PCR方法扩增一段 1.3kb的片段cDNA ;将此片段插入pUC19质粒载体。结果 克隆了丙型肝炎基因组非结构NS4区全长基因 ,构建了pUC -NS4重组体 ,对重组体进行酶切鉴定 ,克隆的cDNA与预计的片段大小相符 ,并作序列测定。结论 获得全长NS4区基因克隆 ,经序列分析表明 ,与日本HC -J6有较高的同源性 (92 .1%)。 展开更多
关键词 限制性片段长度多态性分析 聚合酶链反应 克隆 测序 转录重组体 克隆 基因扩增 HCV 2a/Ⅲ肝炎病毒
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手术室医护人员乙型肝炎病毒感染的职业防护
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作者 李连华 郭丽娟 马文秀 《吉林医学》 CAS 2006年第8期905-905,共1页
关键词 手术室 2型肝炎病毒感染 职业防护
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徐州市育龄期成人乙型肝炎流行病学特征分析 被引量:2
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作者 张先梅 郭静 《河北医药》 CAS 2017年第3期458-459,462,共3页
目的了解本地区育龄期人群乙型肝炎病毒的感染及流行情况,为今后乙肝防治工作提供参考依据。方法回顾性分析体检、就诊的1 318例育龄期成人HBV血清学标志物(采用磁微粒化学发光酶免疫分析法检测)。结果育龄期人群有13种HBV感染模式,以单... 目的了解本地区育龄期人群乙型肝炎病毒的感染及流行情况,为今后乙肝防治工作提供参考依据。方法回顾性分析体检、就诊的1 318例育龄期成人HBV血清学标志物(采用磁微粒化学发光酶免疫分析法检测)。结果育龄期人群有13种HBV感染模式,以单独HBs Ab阳性和全阴性两种模式为主,分别占49.69%和31.41%;感染模式不存在性别分布差异;非正常妊娠人群与正常妊娠人群乙肝病毒感染模式分布差异无统计学意义(P>0.05)。结论本地区育龄期人口乙肝易感人群比重较大,约占1/3,需积极进行乙肝疫苗接种;非正常妊娠可能与HBV感染无关。 展开更多
关键词 育龄期 2型肝炎病毒 血清学标志物
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酵母表达的重组戊型肝炎病毒ORF2蛋白在实验感染恒河猴血清抗体检测中的应用 被引量:6
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作者 鲁健 佟玉品 +1 位作者 江永珍 毕胜利 《中华实验和临床病毒学杂志》 CAS CSCD 北大核心 2001年第4期382-383,共2页
关键词 酵母 表达 重组戊肝炎病毒ORF2蛋白 血清抗体 检测 实验感染
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抗戊型肝炎病毒E2 IgM诊断急性戊型肝炎的敏感性和特异性 被引量:9
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作者 郑玲 柳丽娟 +6 位作者 胡莹盈 王艳丽 温珠妹 张宇 陈惠聪 张启云 江家骥 《中华肝脏病杂志》 CAS CSCD 北大核心 2005年第8期590-593,共4页
目的评价抗戊型肝炎病毒(HEV)衣壳蛋白重组抗原E2 IgM(抗-E2 IgM)诊断急性散发性戊型肝炎的敏感性和特异性。方法用酶联免疫吸附法检测176份急性散发性戊型肝炎和191份急性散发性非甲~非戊型肝炎患者血清中抗-E2 IgM,与国产传统试剂和... 目的评价抗戊型肝炎病毒(HEV)衣壳蛋白重组抗原E2 IgM(抗-E2 IgM)诊断急性散发性戊型肝炎的敏感性和特异性。方法用酶联免疫吸附法检测176份急性散发性戊型肝炎和191份急性散发性非甲~非戊型肝炎患者血清中抗-E2 IgM,与国产传统试剂和新加坡Genelabs试剂检测的IgM(GL-IgM)作比较;对抗-E2 IgM阳性血清检测血清中HEV RNA,采用logistic回归分析检测抗-E2 IgM和HEV RNA的相关因素。结果在176份急性戊型肝炎患者血清中,抗-E2 IgM的检出率为68.75%,国产传统试剂抗-HEV IgM检出率为56.25%,X2IgM=6.49,P<0.05。在191份急性非甲~非戊型肝炎血清中有37例(19.37%)抗- E2 IgM阳性,其中11例GL-IgM同时阳性;在158份抗-E2 IgM阳性血清中,有81例HEV RNA阳性(51.27%), 其中急性戊型肝炎的阳性率为57.02%,急性非甲~非戊型肝炎的阳性率为32.43%,23例抗-E2 IgM阴性的急性戊型肝炎患者的血清,无一例检测到HEV RNA。Logistic多因素回归分析发现,抗-E2 IgM的检出率与发病至入院时间、年龄、血清胆红素、血清氨基转移酶水平无关,血清丙氨酸氨基转移酶水平与HEV RNA水平呈正相关(P=0.024)。结论抗-E2 IgM是HEV急性期感染敏感性和特异性强的血清学指标;HEV感染仍是部分临床诊断为急性非甲~非戊型肝炎的病因;持续HEV病毒血症可能是影响急性戊型肝炎病情的重要因素。 展开更多
关键词 抗戊肝炎病毒E2 IGM 急性戊肝炎 敏感性 特异性 肝炎抗体 血清学
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2a型丙型肝炎病毒5′非编码区的RACE扩增及二级结构的分析
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作者 秦兆习 丛旭 +3 位作者 蒋栋 哈明昊 陈红松 魏来 《中华实验和临床病毒学杂志》 CAS CSCD 北大核心 2002年第4期333-336,共4页
目的 获得真全长中国大陆 2a型丙型肝炎病毒 (HCV) 5′非编码区 (5′UTR)cDNA ,并分析其一级结构和二级结构 ,为进一步研究其在HCV复制、翻译中的调控机制、开发新的抗HCV药物奠定基础。方法 利用逆转录套式聚合酶链反应 (RT PCR)联... 目的 获得真全长中国大陆 2a型丙型肝炎病毒 (HCV) 5′非编码区 (5′UTR)cDNA ,并分析其一级结构和二级结构 ,为进一步研究其在HCV复制、翻译中的调控机制、开发新的抗HCV药物奠定基础。方法 利用逆转录套式聚合酶链反应 (RT PCR)联合限制性内切酶长度多态性分析(RFLP)初步筛选出 1例 2a型HCV感染者 ,采用cDNA末端快速扩增技术 (RACE)扩增出一条约 80 0bp的cDNA片段 ,A T克隆 ,用RFLP与PCR鉴定重组子 ,全自动序列分析仪测定插入子序列 ,RNAdraw预测二级结构。结果 RACE法获得真全长 2a型HCV 5′端序列。 5个克隆中 ,3个克隆含全长 5′UTR序列 ,与HCV 1,HC C2 ,HC J6 ,HC J8相比 ,同源性分别为 93.6 %~ 94 .4 % ,92 .1%~ 93.0 % ,98.8%~99 .7% ,96 .2 %~ 96 .5 % ,与 2a型标准株HC J6相比 ,2 1,170 ,2 2 2 ,2 4 7,339位不同 ,分别为G ,A ,C ,C ,T ,但这些突变不影响其二级结构。另外 2个克隆为 5′端缺失突变株 ,分别缺失 5 4bp和 14 4bp。结论 RACE技术快速、有效、实用 ,可有效获得病毒基因组的末端序列 ;依此获得中国大陆 2a型HCV的 5′UTRcDNA ;在感染者血液中存在 5′端部分缺失的HCV基因片段。 展开更多
关键词 2a肝炎病毒 二级结构 肝炎 5'非编码区 序列分析 CDNA末端快速扩增技术 一级结构
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Association between hepatocellular carcinoma and type 2 diabetes mellitus in Italy:Potential role of insulin 被引量:11
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作者 Valter Donadon Massimiliano Balbi +2 位作者 Pietro Casarin Alessandro Vario Alfredo Alberti 《World Journal of Gastroenterology》 SCIE CAS CSCD 2008年第37期5695-5700,共6页
AIM: To investigate the relationships between Type 2 diabetes mellitus (DM2) and the risk of hepatocellular carcinoma (HCC). METHODS: We studied the association between DM2 and HCC in a large case-control study that e... AIM: To investigate the relationships between Type 2 diabetes mellitus (DM2) and the risk of hepatocellular carcinoma (HCC). METHODS: We studied the association between DM2 and HCC in a large case-control study that enrolled 465 consecutive Caucasian patients with HCC (78.3% males, mean age 68.5 ± 8.9 years) compared with an age and sex matched control group of 490 subjects. RESULTS: Prevalence of DM2 was significantly higher in HCC patients (31.2% vs 12.7%; OR = 3.12, 95% CI: 2.22-4.43) and in HCC cases with alcohol abuse. DM2 has been diagnosed before the appearance of HCC in 84.1% of diabetic HCC subjects with mean duration of 141.5 mo, higher in cases treated with insulin than in those with oral antidiabetic agents (171.5 vs 118.7 mo). Compared to controls, males DM2 with HCC were more frequently treated with insulin (38.1% vs 17.6%, P = 0.009) and with sulfonylurea with or without metformin than with diet with or without metformin (84% vs 68.3%, P = 0.049). CONCLUSION: DM2 in our patients is associated with a 3-fold increase risk of HCC. In most of our cases DM2 pre-existed to HCC. Patients with DM2 and chronic liver disease, particularly insulin treated males, should be considered for HCC close surveillance programs. 展开更多
关键词 Hepatocellular carcinoma Type 2 diabetesmellitus Hepatitis virus B and C INSULIN Antidiabetic therapy
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Relationship between hepatitis C virus infection and type 2 diabetes mellitus:Meta-analysis 被引量:14
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作者 Cho Naing Joon Wah Mak +1 位作者 Syed Imran Ahmed Mala Maung 《World Journal of Gastroenterology》 SCIE CAS CSCD 2012年第14期1642-1651,共10页
AIM:To investigate the association between hepatitis C infection and type 2 diabetes mellitus.METHODS:Observational studies assessing the relationship between hepatitis C infection and type 2 diabetes mellitus were id... AIM:To investigate the association between hepatitis C infection and type 2 diabetes mellitus.METHODS:Observational studies assessing the relationship between hepatitis C infection and type 2 diabetes mellitus were identified via electronic and hand searches.Studies published between 1988 to March 2011 were screened,according to the inclusion criteria set for the present analysis.Authors performed separate analyses for the comparisons between hepatitis C virus(HCV) infected and not infected,and HCV infected and hepatitis B virus infected.The included studies were further subgrouped according to the study design.Heterogenity was assessed using I2 statistics.The summary odds ratios with their corresponding 95% CIs were calculated based on a random-effects model.The included studies were subgrouped according to the study design.To assess any factor that could potentially affect the outcome,results were further stratified by age group(proportion of ≥ 40 years),gender(proportion of male gender),body mass index(BMI)(pro-portion of BMI ≥ 27),and family history of diabetes(i.e.,self reported).For stability of results,a sensitivity analysis was conducted including only prospective studies.RESULTS:Combining the electronic database and hand searches,a total of 35 observational studies(in 31 articles) were identified for the final analysis.Based on random-effects model,17 studies(n = 286 084) compared hepatitis C-infected patients with those who were uninfected [summary odds ratio(OR):1.68,95% CI:1.15-2.45].Of these 17 studies,7 were both a cross-sectional design(41.2%) and cohort design(41.2%),while 3 were case-control studies(17.6%).Nineteen studies(n = 51 156) compared hepatitis C-infected participants with hepatitis B-infected(summary OR:1.92,95% CI:1.41-2.62).Of these 19 studies,4(21.1%),6(31.6%) and 9(47.4%) were cross-sectional,cohort and case-control studies,respectively.A sensitivity analysis with 3 prospective studies indicated that hepatitis C-infected patients had a higher risk of developing type 2 diabetes compared with uninfected controls(summary odds ratio:1.41,95% CI:1.17-1.7;I2 = 0%).Among hepatitis C-infected patients,male patients(OR:1.26,95% CI:1.03-1.54) with age over 40 years(summary OR:7.39,95% CI:3.82-9.38) had an increased frequency of type 2 diabetes.Some caution must be taken in the interpretation of these results because there may be unmeasured confounding factors which may introduce bias.CONCLUSION:The findings support the association between hepatitis C infection and type 2 diabetes mellitus.The direction of association remains to be determined,however.Prospective studies with adequate sample sizes are recommended. 展开更多
关键词 Hepatitis C Type 2 diabetes mellitus Observational studies META-ANALYSIS
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合欢饮的水提物体外抗HCV病毒活性研究 被引量:1
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作者 巫园园 梁泳淋 +1 位作者 陈飞龙 汤庆发 《海南医学院学报》 CAS 2022年第1期1-5,共5页
目的:研究合欢饮的水提物抗HCV病毒活性及其机制。方法:采用Huh7.5.1细胞建立HCV2a病毒感染模型;通过CellTiter-Glo法测定吸光度值计算得到细胞存活率(%)和RenillaLuciferaseAssayKit法测定Rluc值算得到抑制率(%)、CC_(50)、EC_(50)并计... 目的:研究合欢饮的水提物抗HCV病毒活性及其机制。方法:采用Huh7.5.1细胞建立HCV2a病毒感染模型;通过CellTiter-Glo法测定吸光度值计算得到细胞存活率(%)和RenillaLuciferaseAssayKit法测定Rluc值算得到抑制率(%)、CC_(50)、EC_(50)并计算SI值;通过检测荧光素酶活性观察病毒72 h内对不同浓度合欢饮的耐药性;应用Western-blot检测NS5A、NS3及NS5B蛋白表达水平。结果:合欢饮水提物抗HCV2a的CC_(50)、EC_(50)及SI分别为132.50、1.90、67.90μg/mL。在24、48、72 h给药后的EC_(50)分别为18.0、5.8、2.3μg/mL。在给药浓度范围内,合欢饮水提物对NS5A及NS5B蛋白的表达均有抑制作用且呈一定的量效关系,但对NS3蛋白表达的作用不明显。结论:合欢饮的水提物可能通过改变NS5A及NS5B蛋白表达水平来抑制HCV2a病毒的复制,且长时间给药病毒无耐药性。 展开更多
关键词 合欢饮水提物 肝炎2a病毒 体外实验 病毒活性
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Treatment of genotype 2 and 3 chronic hepatitis C virus-infected patients
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作者 Perdita Wietzke-Braun Volker Meier +2 位作者 Katrin Neubauer-Saile Sabine Mihm Giuliano Ramadori 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第39期6188-6192,共5页
AIM: Before pegylated interferon alpha (IFN) was introduced for the therapy of chronic hepatitis C virus (HCV)-induced hepatitis, conventional thrice weekly IFN therapy was supplemented by ribavirin. Also, at tha... AIM: Before pegylated interferon alpha (IFN) was introduced for the therapy of chronic hepatitis C virus (HCV)-induced hepatitis, conventional thrice weekly IFN therapy was supplemented by ribavirin. Also, at that time, higher and more frequent doses of IFN were expected to be more effective than the standard regimen of 3 MU thrice weekly. As ribavirin significantly increases side effects and negatively influences the quality of life particularly in young patients, we started a prospective non-randomized study with a daily IFN-2a monotherapy as an initial treatment for chronic hepatitis C. METHODS: Forty-six consecutive chronic HCV-infected patients received 3 MU IFN-2a per day as an initial treatment. Patients with genotype 2 or 3 (n = 12) were treated for 24 wk, and patients with genotypes other than 2 or 3 (n = 34) for 48 wk. Treatment outcome was followed up for 48 wk after the end of treatment (EOT). Virological response was defined as the absence of detectable serum HCV-RNA. Patients without virological response at 12 wk after the start of treatment received low-dose ribavirin (10 mg(kg·d)) additionally. RESULTS: During treatment, three genotype 3 patients were excluded from the study due to incompliance. The remaining patients (n = 9) infected with genotype 2 or 3 showed an initial virological response rate of 100%. Six patients (66.7%) were still found to be virus-free at the end of follow-up period. In these patients, initial virological response was evident already after 2 wk of treatment. In contrast, initial virological response occurred first after 4 wk of treatment in the three patients who relapsed (33.3%). In comparison, patients infected with genotypes other than 2 or 3 (n = 34) showed an initial virological response rate of only 23.5% (n = 8), and even in combination with ribavirin a sustained virological response (SVR) rate of only 11.8% (n = 4) could be achieved. CONCLUSION: In chronic HCV-infected patients with genotype 2 or 3, a SVR can be expected after 24 wk of daily dose IFN-2a treatment without ribavirin, if initial virological response develops early. This finding is worth to be confirmed in a prospective randomized study with pegylated IFN. 展开更多
关键词 Chronic hepatitis C virus infection Genotype 2 and 3 Alpha interferon Daily dose interferon therapy
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