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丙型肝炎病毒体外感染人肝癌细胞株HepG2的研究 总被引:3,自引:0,他引:3
目的:研究人肝癌细胞株HepG2体外对丙型肝炎病毒(hepatitis C virus,HCV)的易感性。方法:慢性丙型肝炎患者的血清与HepG2共同孵育后,以逆转录-聚合酶链反应、免疫组化法、原位杂交法分别检测细胞和(或)培养上清中的HCV正、负链RNA、HCV抗原表达和HCV RNA在细胞内的定位。结果:感染血清和细胞共同孵育后的第2-40天,从细胞内和培养上清中均可间断地检测出HCV正、负链HCV RNA,HCV NS3抗原在细胞内能稳定表达,原位杂交证实HCV RNA阳性物质多位于细胞质中。结论:HepG2细胞在体外不但对HCV易感,而且可以支持HCV体外复制。 相似文献
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本文综述了近几年丙型肝炎病毒(HCV)特异性细胞免疫研究进展,重点介绍了CD8+细胞毒性T淋巴细胞(CTL)介导的细胞免疫在抗HCV感染中的作用及来自病毒方面的并分析了小鼠在这一研究领域的价值。 相似文献
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李越希 《国外医学:病毒学分册》1998,5(3):68-72
本文综述了丙型肝炎病毒(Hepatitis C Virus,HCV)体外细胞培养研究的最新进展情况,分析了体外细胞培养HCV时需要注意的一些重要因素及目前存在的主要问题。 相似文献
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目的 研究丙型肝炎病毒 (HCV)体外细胞培养方法。方法 原代人胎肝细胞与HCV感染血清共孵育后 ,用逆转录 聚合酶链反应、原位杂交、免疫组化分别检测细胞和培养上清中的HCVRNA和HCV抗原表达。结果 从感染血清和细胞共同孵育后的 2~ 2 5d ,细胞内和 /或培养上清中可间断检出HCV正、负链RNA ;HCVNS3抗原在细胞内能稳定表达 ;原位杂交显示HCV负链RNA阳性物质多位于细胞浆。结论 原代人胎肝细胞对HCV易感 ,可用作HCV体外感染和复制的靶细胞 相似文献
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丙型肝炎病毒结构蛋白在痘苗病毒中的表达 总被引:1,自引:0,他引:1
为研究中国丙型肝炎病毒(HCV)的抗原性及在细胞内的加工,将丙型肝炎病毒(HCV)5’非编码区(NTR)和结构基因(Core+E1+E2/NS1)插入痘苗病毒表达载体pJSA1175中,转染TK-143细胞,经纯化得到丙型肝炎(HCV)重组痘苗病毒vJSA1175CE株。Southernblot杂交表明,HCV结构基因存在于痘苗病毒之中。Westernblot分析发现,vJSA1175CE表达蛋白带位于90kDa,为一多聚蛋白;此蛋白为分泌型,分泌量与细胞裂解物内量大致相同 相似文献
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李聚林 《国外医学:病毒学分册》2006,(6)
丙型肝炎在全世界广泛流行,并且有不少地区还表现出上升的趋势,严重程度及其复杂性日益突出,使采供血工作以及输血安全面临许多问题,所以本文重点就丙型肝炎病毒与安全血液的若干问题作一综述。 相似文献
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丙型肝炎病毒基因分型及其与疾病程度、感染途径和干扰素疗效的关系 总被引:9,自引:0,他引:9
为探讨广州地区丙型肝炎病毒(HCV)感染的基因型及其与疾病程度、感染途径和干扰素疗效的关系,作者应用分型PCR技术对156例抗-HCV阳性患者血清HCVRNA进行分型检测,并分析不同程度肝病及不同感染途径HCV基因型分布,且对51例经干扰素治疗的慢性丙肝作分型评价。结果124例HCVRNA阳性中,Ⅱ型占90.3%(112例),Ⅲ型占8.1%(10例),Ⅱ/Ⅲ型混合感染占1.6%(2例),未检出Ⅰ、Ⅳ型,说明广州地区HCV感染以Ⅱ型为主;不同程度肝病及不同感染途径之间HCV基因型构成比无明显差异,表明基因型与疾病严重程度及感染途径关系不大;干扰素治疗病例,HCV-Ⅱ、Ⅲ型感染的总应答数、完全应答数及部分应答数分别为18/41,11/41,7/41和8/10,6/10,2/10,提示Ⅲ型感染疗效较好,HCV基因型似可作为选择干扰素治疗病例及判断疗效的参考指标,但是由于观察的Ⅲ型病例数较少,需积累更多的资料才能作出更客观的结论。 相似文献
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目的探讨丙型肝炎患者血清丙型肝炎病毒抗体(抗-HCV)结果与丙型肝炎病毒核糖核酸(HCV—RNA)定量的相关性。方法采用酶联免疫吸咐实验(ELISA)和实时荧光定量PCR(FQ—PCR)技术,对621例同时做抗-HCV和HCV—RNA的丙型肝炎患者检测。结果621份标本中有115份HCV—RNA含量高于1000拷贝/ml,124份抗-HCV阳性。经统计分析,两者有明显的相关性。结论血清抗-HCV与血清HCV—RNA之间有较好的一致性;抗-HCV检测和HCV—RNA检测均有一定的局限性,同时检测抗-HCV和HCV-RNA可提高丙型肝炎患者的检出率.为其诊断和治疗提供帮助。 相似文献
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献血员血清中抗丙型肝炎病毒抗体检测 总被引:1,自引:0,他引:1
献血员血清中抗丙型肝炎病毒抗体检测谭德明,胡国龄,刘双虎,张铮有马晖勝1989年Choo报道了检测血清抗丙型肝炎病毒抗体(以下简称抗-HCV)的方法[1,2],发现输血后非甲非乙型肝炎中80%~90%为丙型肝炎病毒所致[2,3]。为了解丙型肝炎病毒感... 相似文献
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乙型肝炎病毒阳性血清体外感染Hep G2细胞的实验研究 总被引:2,自引:1,他引:2
目的 建立HBV阳性血清体外感染HepG2 细胞的实验方法。方法 培养HepG2 细胞传至6孔板中,2 4h后进行HBV阳性血清体外感染HepG2 细胞的实验。感染组用HBV阳性血清,阴性对照组用HBV阴性血清,空白对照组用DMEM培养基。实验开始后HepG2 细胞继续孵育2 4h ,而后用0 0 1mol LPBS清洗8次后加入2 %DMEM培养液。收集PBS第8次洗液,收集PBS洗后每隔12h各孔细胞培养上清。ELISA检测细胞培养上清中的HBsAg。PCR检测细胞培养上清和HepG2 细胞中的HBVDNA。结果 感染组在PBS洗后12h的细胞培养上清中ELISA检测HBsAg呈阳性。PCR检测显示感染组细胞培养上清和HepG2 细胞中HBVDNA呈阳性,阴性对照组和空白对照组HBVDNA呈阴性。结论 HBV阳性血清进行HBV感染体外培养HepG2 细胞是可行的。 相似文献
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《Clinical microbiology and infection》2018,24(4):422-427
ObjectivesTo analyse the variation of hepatitis C virus (HCV) prevalence and genotype distribution and their determinants in people living with human immunodeficiency virus (HIV) who entered care between 1997 and 2015.MethodsHIV-infected patients enrolled in ICONA who were tested for HCV antibodies (HCV-Ab) were included.ResultsOverall 3407 of 12 135 (28.1%) were HCV-Ab+; and 735 of 12 135 (6.1%) were HBsAg+. Among patients whose HCV genotype was known, the most represented were genotypes 1 and 3. The prevalence of HCV infection decreased from 49.2% (2565/5217) during 1997–2002 to 10.2% (556/5466) during 2009–2015. The frequency of genotype 1a increased from 29.0% (264/911) to 43.0% (129/300), whereas genotype 3 decreased from 38.5% (351/911) to 27.0% (81/300). Independent predictors of HCV-Ab+ status were being female (adjusted OR (AOR) 1.23, 95% CI 1.04–1.50, p = 0.01), risk category (versus injecting drug users: men who have sex with men AOR 0.01, 95% CI 0.01–0.01, p <0.001; heterosexuals AOR 0.01, 95% CI 0.01–0.01, p <0.001; other/unknown AOR 0.02, 95% CI 0.01–0.02, p <0.001), being cared for in Central Italy (versus being cared for in Northern Italy: AOR 0.85, 95% CI 0.73–0.98, p <0.001), being Italian-born (AOR 1.44, 95% CI 1.16–1.80, p = 0.001) and being enrolled in less recent calendar years (versus 1997–2002: 2009–2015 AOR 0.23, 95% CI 0.19–0.27, p <0.001; 2003–2008 AOR 0.49, 95% CI 0.41–0.61, p <0.001).ConclusionsThe prevalence of HCV infection in HIV-infected patients entering into care in Italy significantly declined in more recent calendar years. After adjusting for risk factors and calendar years, HCV co-infection was more frequent in females and in those born in Italy. 相似文献
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丙型肝炎患者肾小球病变的观察 总被引:2,自引:0,他引:2
为了明确丙型肝炎病毒(HCV)感染与肾脏疾病的关系,采用免疫组化技术和抗-HCVNS3和抗-HCVNS5单克隆抗体,检测了21例丙肝患尸解肾组织中的HCAg,结果13例阳性(61.9%),其中11例为膜增殖性肾炎(MPGN),1例为系膜增殖性肾炎(MsPGN),1例肾组织大致正常。8例HCAg检测阴性,其中膜性肾病(MN)1例,膜增殖性肾炎2例,毛细血管内增殖性肾炎1例,余4例肾组织未见明显异常。与乙型肝炎对照组(33.0%)相比,丙肝患者肾脏病变更为多见,形态多样,以MPGN为主。同时对丙肝患者有肾脏病变者的临床资料进行了分析,病人可表现为高血压,蛋白尿,血尿及早期血清尿素氮(BUN)增高,因此认为同乙型肝炎病毒(HBV)感染后相同,也存在着丙肝相关性肾炎 相似文献
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The subversion of immune responses that hepatitis C virus (HCV) uses to escape immune surveillance and to establish persistent infection has been poorly understood. The immune-suppressive molecule human leukocyte antigen-G (HLA-G) has been supposed to play important roles in viral infection. In the current study, HCV genotype was analyzed in 67 chronic HCV-infected (CHC) patients. Plasma soluble sHLA-G (including sHLA-G1 and HLA-G5), interleukin-10 (IL-10), and interferon-γ (IFN-γ) levels were determined in these CHC patients and in healthy subjects by enzyme-linked immunosorbent assay, and the sHLA-G isoforms present in plasma were determined by Western blot. Data showed that HCV 1b was the predominant genotype, with a prevalence of 64.2%. sHLA-G was dramatically increased in CHC patients (median: 85.54 U/ml, range: 19.40-204.07) over that in normal controls (median: 9.13 U/ml, range: 5.07-69.56) (p < 0.001). Western blotting revealed that plasma sHLA-G was derived from sHLA-G1 and HLA-G5. IL-10 and IFN-γ levels were also significant higher in CHC patients than in normal controls (median: 16.3 pg/ml vs 1.8 pg/ml, p < 0.001, and 1025.3 pg/ml vs 858.3 pg/ml, p = 0.03, respectively). No significant association was observed for the HCV genotype and viral RNA load with the levels of sHLA-G, IL-10, and IFN-γ in CHC patients. These results indicate that elevation of sHLA-G expression in HCV patients was independent of viral genotype and viral RNA load. Given its immunotolerant property, an increase in sHLA-G may play a role in the persistency of HCV infection. 相似文献
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丙型肝炎病毒感染的血清学检测 总被引:2,自引:0,他引:2
目的了解丙型肝炎病毒(HCV)感染及病毒血症存在情况。方法用酶联免疫吸附试验(ELISA)和聚合酶链反应(PCR)对不同人群的血清标本做了抗-HCV、抗-HCVIgM和HCVRNA的检测,并对三项指标间的关系进行了对比分析。结果抗-HCV在普通成年人、献血员、急性肝炎和肝硬化患者中的检出率分别是357%,858%,625%和4838%;与HCVRNA的符合率分别是1143%,6111%,800%和7333%。相同人群抗-HCVIgM与HCVRNA的符合率分别是75%,909%,8181%和100%。结论抗-HCVIgM比抗-HCV能更客观地反映HCV病毒血症的情况,个别抗-HCV阴性血清检测到了抗-HCVIgM和HCVRNA。 相似文献
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Jae Young Jang Soung Won Jeong Sung Ran Cheon Sae Hwan Lee Sang Gyune Kim Young Koog Cheon Young Seok Kim Young Deok Cho Hong Soo Kim So Young Jin Yun Soo Kim Boo Sung Kim 《Clinical and molecular hepatology》2011,17(3):206-212
Background/Aims
We investigated the frequency of occult hepatitis B virus (HBV) infection in anti-hepatitis C virus (HCV)-positive individuals and the effects of occult HBV infection on the severity of liver disease.Methods
Seventy-one hepatitis B virus surface-antigen (HBsAg)-negative patients were divided according to their HBV serological status into groups A (anti-HBc positive, anti-HBs negative; n=18), B (anti-HBc positive, anti-HBs positive; n=34), and C (anti-HBc negative, anti-HBs positive/negative; n=19), and by anti-HCV positivity (anti-HCV positive; n=32 vs. anti-HCV negative; n=39). Liver biopsy samples were taken, and HBV DNA was quantified by real-time PCR.Results
Intrahepatic HBV DNA was detected in 32.4% (23/71) of the entire cohort, and HBV DNA levels were invariably low in the different groups. Occult HBV infection was detected more frequently in the anti-HBc-positive patients. Intrahepatic HBV DNA was detected in 28.1% (9/32) of the anti-HCV-positive and 35.9% (14/39) of the anti-HCV-negative subjects. The HCV genotype did not affect the detection rate of intrahepatic HBV DNA. In anti-HCV-positive cases, occult HBV infection did not affect liver disease severity.Conclusions
Low levels of intrahepatic HBV DNA were detected frequently in both HBsAg-negative and anti-HCV-positive cases. However, the frequency of occult HBV infection was not affected by the presence of hepatitis C, and occult HBV infection did not have a significant effect on the disease severity of hepatitis C. 相似文献18.
BackgroundPediatric cancer patients who received blood transfusions were potentially exposed to hepatitis C virus (HCV) prior to second-generation HCV screening of blood products in 1992. Limited evidence is available about long-term incident cirrhosis in this population.ObjectivesWe aimed to estimate the overall and sex-specific incidence of cirrhosis among HCV-seropositive survivors of pediatric cancer.Study designWe identified 113HCV-seropositive pediatric cancer patients treated at St. Jude Children’s Research Hospital between 1962 and 1997, who survived ≥5 years post-diagnosis, and were followed through 2014. Our outcome was cirrhosis determined by liver biopsy or diagnostic imaging. We used a competing-risk framework to estimate the overall and sex-specific cumulative incidence and 95% confidence limits (CL) of cirrhosis at 10-year follow-up intervals.ResultsThe median duration of follow-up was 30 years (interquartile range = 28–36) post-cancer diagnosis. Cumulative incidence of cirrhosis increased at each 10-year interval from 0% after 10 years to 13% after 40 years (Ptrend < 0.001). The median age at diagnosis of cirrhosis was 30 years (interquartile range = 24–38). We observed a linear trend in incidence for males (Ptrend < 0.001), with a cumulative incidence of 18% (95% CL: 6.1%, 34%) after 40 years. The cumulative incidence for females was 6.5% (95% CL: 0.42%, 26%) after 40 years, but we did not observe a linear trend (Ptrend = 0.99).ConclusionOur results suggest that the incidence of cirrhosis is similar between HCV-seropositive pediatric cancer survivors and the general population given similar duration of follow-up, but survivors may be diagnosed with cirrhosis at an earlier age. 相似文献