用基因重组丙型肝炎病毒抗原C11和C7对丙型肝炎进行分析

使用基因重组丙型肝炎病毒（ＨＣＶ）基因核心区所表达的抗原Ｃ１１及非结构区ＮＳ３区所表达的抗原Ｃ７，对北京地区慢性病毒性肝炎、肝硬化、原发性肝瘤４４２例进行了血清丙型肝炎病毒抗体（抗ＨＣＶ）检测。初步看出本地区乙型肝炎更多于丙型肝炎。肝硬变、原发性肝癌在病因学上与乙、丙型肝炎重叠感染的关系比单独乙型或单独丙型肝炎感染的关系更为密切。提示这些患者中乙型及丙型肝炎病毒之间有相互作用的可能性。     

HCV genotypes in hepatitis C patients and their clinical significances

ＩＮＴＲＯＤＵＣＴＩＯＮＩｔｈａｓｂｅｅｎｄｉｓｃｏｖｅｒｅｄｔｈａｔｈｅｐａｔｉｔｉｓＣｖｉｒｕｓ（ＨＣＶ）ｐｒｅｓｅｎｔｓｃｏｎｓｉｄｅｒａｂｌｅｎｕｃｌｅｏｔｉｄｅｖａｒｉａｔｉｏｎａｎｄｈａｓｍａｎｙｇｅｎｏｔｙｐｅｓ．Ａｔｌｅａｓｔｔｗｅｌｖｅ,５ｏｆｗｈｉｃｈｐｒｅｖａｌｅｎｔ,ｉ．ｅ．：Ⅰ／１ａ,Ⅱ／１ｂ,Ⅲ／２ａ,Ⅳ／２ｂ,ａｎｄⅤ／３ａｔｙｐｅｓ．ＴｈｅｄｉｆｆｅｒｅｎｔｇｅｎｏｔｙｐｅｓｏｆＨＣＶｍａｙｐｏｓｓｅｓｓｓｏｍｅｒｅｔａｔｉｏｎｓｈｉｐｗｉｔｈｒｅｇｉｏｎａｌｄｉ…     

急性丙型肝炎基因分型的临床分析

目的探讨丙型肝炎病毒基因分型与感染方式、肝功能损伤程度的关系及干扰素疗效与基因型关系。方法采用５′－末端非编码区酶切基因分型技术检测血清ＨＣＶＲＮＡ基因型。结果有受血史患者以Ⅱ型ＨＣＶ感染为主，Ⅲ型多见于散发病例，Ｐ＜０．０５；Ⅱ型肝功能损伤重，Ｐ＜０．０５；Ⅲ型ＨＣＶＲＮＡ干扰素疗效好于Ⅱ型。结论肝功能损伤程度与基因分型有关，输血机会的增加使感染Ⅱ型ＨＣＶＲＮＡ机会增加，急性期干扰素抗病毒治疗有效     

慢性丙型肝炎病毒感染者外周血淋巴细胞增殖反应

目的　观察慢性丙型肝炎患者外周血淋巴细胞对丙型肝炎病毒（ＨＣＶ） 抗原刺激的增殖反应．方法　外周血单个核细胞（ＰＢＭＣ） 与ＨＣＶ 抗原ｃ２２ 、ｃ３３ 、ｃ１００ － ３ 、ＮＳ５ 和植物血凝素（ＰＨＡ） 分别共同孵育,加入胸腺嘧啶核苷（３ ＨＴｄＲ） ,然后收集细胞于液闪仪测定每分钟脉冲数（ｃｐｍ） ．结果　根据对不同ＨＣＶ 抗原的淋巴细胞增殖反应发现,以ｃ２２免疫原性最强,ｃ１００ － ３ 次之：淋巴细胞激活与ＨＣＶ 基因型关系不大;健康对照和慢性乙型肝炎患者对各ＨＣＶ 抗原未能显示有效的淋巴细胞增殖反应;与健康对照比较,慢性丙型肝炎和乙型肝炎患者对ＰＨＡ 刺激的淋巴细胞增殖反应降低．结论　ＨＣＶ 抗原ｃ２２ 免疫原性最强,丙型肝炎患者对ＨＣＶ 抗原的淋巴细胞增殖反应系特异性;慢性丙型肝炎和乙型肝炎患者存在抑制的细胞免疫应答．     

广东省单纯丙型肝炎病毒及其与人类免疫缺陷病毒共感染者丙型肝炎病毒的基因亚型分析

目的 了解广东省人类免疫缺陷病毒(HW) /HCV共感染者与单纯HCV感染者的感染途径、HCV的基因亚型分布及其遗传特征,为丙型肝炎病毒感染的治疗与预防提供依据. 方法采用巢式逆转录-聚合酶链反应扩增广东省95例HIV/HCV共感染者及99例单纯HCV感染者HCVNS5B基因区域,扩增产物测序后进行HCV基因亚型分析,遗传分析利用MEGA4软件.不同种基因亚型型内基因距离以碱基置换率表示.结果 HIV/HCV共感染者HCV有5种基因亚型,其中6a型占53.7％ (51/95)、3a型占17.9％ (17/95)、1b型占15.8％ (15/95)、3b型占11.6％ (11/95)、1a型占1.0％(1/95);1b亚型内基因距离为6.30％±1.27％,高于其他基因亚型.HIV/HCV共感染者主要通过静脉注射毒品感染(75/95,78.9％),其基因亚型主要为6a型,占60.0％ (45/75).单纯HCV感染者HCV有7种基因亚型,其中1b型占67.7％ (67/99)、6a型占17.2％(17/99)、3a型占6.1％ (6/99)、2a型占5.％(5/99)、3b型占2.0％(2/99)、4a型占1.0％ (1/99)、5a型占1.0％(1/99);1b亚型内基因距离为5.17％±1.03％,高于其他基因亚型.单纯HCV感染者主要通过输血或血液制品感染(80/99,80.8％),其HCV基因亚型主要为1b型,占76.2％ (61/80).结论 广东地区HIV/HCV共感染者及单纯HCV感染者HCV基因亚型呈现多样性,HIV/HCV共感染者与单纯HCV感染者主要感染途径不同,HCV主要基因亚型也不同.     

沈阳地区650例慢性丙型肝炎病毒感染者的病毒基因型分析

HCV感染呈全球分布,不同基因型和亚型的地理分布,可能与抗HCV治疗的疗程及效果相关.本研究以核酸序列分析法对650例患者进行HCV基因型检测分析,以探讨沈阳地区感染人群中HCV的基因型特征. 一、资料与方法     

丙型肝炎患者胃粘膜细胞中HCV NS3、NS5抗原定位研究

目的了解丙型肝炎病毒（ＨＣＶ）在胃粘膜细胞中的感染状况。方法采用免疫组化Ｓ－Ｐ法以抗－ＨＣＶＮＳ３和抗－ＨＣＶＮＳ５检测了３３例慢性丙型肝炎（ＣＨＣ）患者胃粘膜组织中ＨＣＶＮＳ３及ＨＣＶＮＳ５抗原。结果ＨＣＶＮＳ３、ＨＣＶＮＳ５的检出率分别为３３．３３％（１１／３３）和２７．２７％（９／３３）。阳性物质沉积于胃粘膜上皮腺体细胞胞浆，呈单个或片灶状分布，部分病例淋巴细胞胞浆中也可见到。结论胃粘膜细胞中ＨＣＶ抗原的检出，表明ＨＣＶ可能是一种泛嗜性病毒，其机制及生物学意义须进一步研究。     

Hepatitis C virus (HCV) genotypes in Spanish patients with HCV infection: relationship between HCV genotype 1b, cirrhosis and hepatocellular carcinoma

Background/Aims: The influence of the infecting virus genotype on the progression of the underlying liver disease in patients with chronic hepatitis C virus (HCV) infection remains controversial. The aim of this study was to investigate the prevalence of HCV genotypes in Spanish patients with chronic HCV infection and to elucidate the relationship between the infecting genotype and severity of the disease.Methods: A cross-sectional, retrospective analysis of frequency distribution of HCV genotypes was carried out in 414 Spanish patients with chronic HCV infection, including 243 patients with asymptomatic or minimally symptomatic chronic hepatitis, 112 patients with cirrhosis and hepatocellular carcinoma and 59 patients with decompensated cirrhosis. HCV genotype was determined by restriction fragment length polymorphisms of the 5′ non-coding region.Results: Infection with HCV genotype 1b was found in 72% of patients with chronic hepatitis and in more than 90% of patients with cirrhosis, with or without hepatocellular carcinoma. Older age, infection with genotype 1b and absence of overt parenteral exposure as a possible source of infection were associated with cirrhosis and hepatocellular carcinoma by univariate analysis and this association was confirmed by regression analysis.Conclusions: HCV genotype 1b is associated with advanced liver disease in our geographical area. However, this may be related to a cohort-effect caused by over-representation of genotype 1b in older patients with more advanced disease, because, in our country, this HCV genotype appeared earlier in time and is therefore associated with more prolonged periods of infection.     

慢性丙型肝炎患者2型糖尿病并发率调查及其基因型特征分析

目的通过调查慢性丙型肝炎患者2型糖尿病并发率及其与所感染丙型肝炎病毒（HCV）基因型的关系。进一步探讨糖尿病是否为丙型肝炎的肝外表现之一。方法采用荧光定量聚合酶链反应和聚合酶链反应一微板核酸杂交酶联免疫技术对308例慢性丙型肝炎、305例慢性乙型肝炎患者进行乙型肝炎病毒、HCV定性．定量检测和HCV基因型分析并比较其与对照人群糖尿病并发率的差异。结果慢性丙型肝炎患者糖尿病并发率为32．79％，明显高于慢性乙型肝炎（9．84％）及对照组（8．39％）。合并糖尿病的慢性丙型肝炎患者血清丙氨酸氨基转移酶及总胆红素水平显著高于未合并糖尿病者，且以1b型HCV的感染率为最高，占40．59％，与未合并糖尿病者相比差异有统计学意义。结论慢性丙型肝炎患者糖尿病并发率高，以1b型多见，且病情相对较重。     

HCV infection and its clinical features in recipients of blood screened for HCV (C100-3) antibody

Abstract After adoption of the anti-hepatitis C virus (C100-3) test, the incidences of definite and suspected cases of post-transfusional hepatitis (PTH) were 3.3% (7/209) and 7.2% (15/209), respectively. Four patients with definite PTH and seven patients with suspected PTH became positive for hepatitis C virus (HCV)-related antibodies or HCV-RNA after transfusion. These cases that became positive for anti-HCV or HCV-RNA showed a peak of alanine aminotransferase (ALT) more than 4 weeks after operation. Only rare cases that showed ALT peaks within 4 weeks after operation became positive for HCV-related antibodies or HCV-RNA. The peak ALT levels in cases showing positive conversion tended to be higher than those in cases showing no conversion. Judging from these results, cases of suspected PTH include those of transient liver disease attributable to surgery as well as clear cases of HCV infection. Thus new diagnostic criteria are required including data on HCV antibodies or HCV-RNA.     

同步检测血浆和外周血单个核细胞中HCV RNA的临床研究

目的探讨联合检测对提高HCV RNA在慢性丙型肝炎患者中检出率的临床意义.方法利用逆转录聚合酶链反应(RT-PCR)技术对583例ALT升高持续1年以上的患者进行同步检测血浆和外周血单个核细胞(PBMC)中的HCV RNA.结果血浆中HCV RNA总检出率为19.2%.PBMC中HCV RNA总检出率为24.5%,HCV RNA在PBMC中的检出率明显高于血浆中的检出率.结论两种方法同时应用能够提高HCV RNA在慢性丙型肝炎患者中的检出率.     

丙型肝炎病毒C和E1区的DNA改组

目的:利用DNA改组技术进行不同亚型的HCVC和E1区的人工进化.方法:首先利用PCR扩增两段具有较高序列同源性的1kb左右的基因片段,两者相比较同源性大于83%.然后将其等量混合,在Mg2 存在的条件下,用DNase Ⅰ切割成50bp左右的小片段.这些小片段在不外加引物的条件下,利用PCR反应进行重聚,再将重聚物经过一轮正常的PCR扩增.结果:获得了与原来片段大小相当的基因片段.结论:这一技术为进一步筛选高活性的HCV C和E1区打下基础,有利于从一组序列同源性程度较高的基因库构建随机嵌合基因,并为改组其他基因家族提供了借鉴.     

The prevalence of hepatitis C virus (HCV) infection in HIV‐positive individuals in the UK – trends in HCV testing and the impact of HCV on HIV treatment outcomes

Summary. We examined the prevalence of hepatitis C virus (HCV) infection among HIV‐positive individuals in the UK, trends in HCV testing and the impact of HCV on HIV treatment outcomes. Trends over time in HCV prevalence were calculated using each patient’s most recent HCV status at the end of each calendar year. Logistic regression was used to identify factors associated with having a HCV antibody test, and Cox regression was used to determine whether HCV status was associated with the time to experiencing an immunological response to highly active antiretroviral treatment (HAART), time to virological response and viral rebound. Of the 31 765 HIV‐positive individuals seen for care between January 1996 and September 2007, 20 365 (64.1%) individuals were tested for HCV, and 1807 (8.9%) had detectable HCV antibody. The proportion of patients in follow‐up ever tested for HCV increased over time, from 782/8505 (9.2%) in 1996 to 14 280/17 872 (79.9%) in 2007. Nine thousand six hundred and sixty‐nine individuals started HAART for the first time in or after January 2000, of whom, 396 (4.1%) were HCV positive. Presence of HCV infection did not affect initial virological response, virological rebound or immunological response. The cumulative prevalence of HCV in the UK CHIC Study is 8.9%. Despite UK guidelines, over 20% of HIV‐positive individuals have not had their HCV status determined by 2007. HCV infection had no impact on HIV virological outcomes or immunological response to HIV treatment. The long‐term impact on morbidity and mortality remain to be determined.     

丙型肝炎肝组织中HCV RNA和HCV NS5抗原表达及意义

目的研究丙型肝炎肝组织中ＨＣＶＲＮＡ与ＨＣＶＮＳ５抗原分布及其与肝组织损伤的关系．方法应用原位杂交和免疫组化方法检测３４例丙型肝炎病毒（ＨＣＶ）感染者肝组织中ＨＣＶＲＮＡ和ＨＣＶＮＳ５抗原表达状况,采用Ｋｎｏｄｅｌ方法评价肝组织病理损伤程度．结果分别有２１例（６１８％）和２４例（７０６％）患者肝组织中检出ＨＣＶＲＮＡ和ＨＣＶＮＳ５抗原．ＨＣＶＲＮＡ阳性信号主要位于肝细胞浆中,在胆管上皮细胞和炎性浸润细胞中也可检出,其分布与肝组织炎症坏死无固定解剖学关系．ＨＣＶＲＮＡ与肝组织中ＨＣＶＮＳ５抗原表达正相关,但二者在肝组织中的分布存在一定差异．多因素分析显示ＨＣＶＮＳ５抗原的表达与肝组织损伤相关．结论丙型肝炎肝组织中ＨＣＶＲＮＡ和ＨＣＶＮＳ５抗原表达相关,病毒在肝组织中的活跃程度与肝损伤有关     

HCV基因型对慢性丙型肝炎干扰素疗效的影响

目的 探讨HCV基因型对慢性丙型肝炎的干扰素（IFN）治疗效果的影响。方法 采用随机、开放和对照的多中心临床试验设计。208例受试者按1：1随机分到聚乙二醇干扰素α—2a(Peg-IFN）组和IFN-α-2a组。在治疗之前，用Simmonds基因分型法酶切分型，在治疗24周结束和完成24周的随访后检测患者的ALT和HCV RNA，以HCV RNA的阴转率作为主要评价指标，经ITT人群的统计学分析。结果 202例患者确定了HCV基因型，基因1型158例（78.2%），非基因1型44例（21.8%），治疗结束病毒应答率（ETVR）和持续病毒应答率（SVR）基因1型患者分别为53.8%和25.3%，非基因1型患者分别为61.4%和43.2%，SVR两组患者差异有显著性，x^2=5.313,P=0.021。Peg IFN组基因1型和非1型患者的ETVR分别为76.8%和81.0%，SVR分别为35.4%和66.7%，SVR两组患者差异有显著性，x^2=6.735,P=0.01。病毒复发率基因1型和非基因1型患者分别为55.6%和23.5%，差异有显著性，x^2=5.496,P=0.02.IFN-α-2a组，ETVR和SVR基因1型患者分别为29.0%和14.5%，非基因1型患者分别43.5%和21.7%，差异无显著性。病毒复发率基因1型患者为72.7%，非基因1型患者为50.0%，差异无显著性。结论 IFN对基因1型丙型肝炎患者的疗效低于非基因1型，HCV基因型主要影响IFN对慢性丙型肝炎的持续应答，也与药物和IFN的疗程相关。     

Hepatitis C virus (HCV) NS5A protein: role in HCV replication and resistance to interferon-alpha

The hepatitis C virus (HCV) non-structural (NS) 5A protein appears to play an important regulatory role on viral replication and could also be involved in viral pathogenesis. HCV resistance to interferon is a complex mechanism involving multiple causes, among which certain NS5A functions could play a role.     

人类免疫缺陷病毒/丙型肝炎病毒混合感染者抗丙型肝炎病毒治疗的进展

由于人类免疫缺陷病毒（HIV）和丙型肝炎病毒（HCV）有相同的传播途径,HIV/HCV混合感染现象十分普遍,已成为严重的公共卫生问题。高效抗逆转录病毒治疗（HAART）的应用显著减少了与HIV感染相关的发病率和病死率,而HCV混合感染引起的慢性肝脏疾病日益成为HIV/HCV混合感染者发病和死亡的重要因素。HIV/HCV混合感染者HCV相关肝病的风险增加,有效的抗HCV治疗对延长这一人群的生存期至关重要。本文就抗HCV治疗对象的评估、治疗时机的选择、治疗的方法、治疗监测和疗效评估以及治疗注意的问题作一综述。     

Optimizing hepatitis C therapy in HIV/hepatitis C virus (HCV) coinfected patients: Analysis of HCV viral kinetics on treatment

INTRODUCTION:

Hepatitis C virus (HCV) infection is potentially curable, but the sustained virological response (SVR) has been shown to be lower in patients coinfected HIV. A single-centre experience treating individuals with HCV and HIV coinfection is reported.

METHODS:

Twenty-one patients who received standard doses of pegylated interferon with weight-based dosing of ribavirin (mean 14.3 mg/kg) were retrospectively reviewed. Qualitative HCV polymerase chain reaction (PCR) was performed prospectively every four weeks if the patient remained HCV PCR positive. All patients with HCV genotype 1 were treated for 48 weeks. Patients with genotype 2 or 3 were treated for 24 weeks and 32 weeks to 36 weeks if their HCV RNA level was undetectable after four weeks (RVR4) or eight weeks (RVR8) of therapy, respectively. If RVR8 was not achieved, the treatment was continued for 48 weeks.

RESULTS:

There were no dropouts or dose reductions within the first 12 weeks of treatment. SVR status was available for 20 patients and adequate serum for viral kinetics analyses was available for 17 patients. Eighty per cent of the patients achieved SVR (50% genotype 1; 100% genotypes 2 and 3). The week 8 viral load remained elevated for all genotype 1 nonresponders.

DISCUSSION:

High effectiveness rates were seen, particularly in patients with HCV genotype 2 and 3 who were treated for shorter durations. HCV viral loads after eight weeks of therapy helped distinguish patients with HCV genotype 1 who would respond to therapy.