丙型肝炎病毒核心抗原在肝细胞癌及癌旁组织中的表达

应用ＳＰ法（链霉菌抗生物素蛋白－过氧化酶连结法）对肝细胞癌（４６例）癌组织及癌旁组织（３８例）中丙型肝炎病毒核心抗原进行免疫组织化学染色，发现两者的检出率分别为２１．７％和３６．８％。阳性染色细胞呈弥漫、灶状和散在分布。抗原定位于肝细胞和肝癌细胞的胞浆内，少数有围核分布的特点。阳性染色多呈细颗粒状，少数呈均质状。癌旁组织抗原表达区域中有淋巴细胞浸润聚集。结果提示丙型肝炎病毒感染在肝细胞癌的发生中可能有一定的关联。     

Expression of HBxAg in human chronic hepatitis,cirrhosis and primary hepatic carcinoma and its significance

The specimens of this study were obtained from 110 cases of chronic hepatitis,108cirrhosis and 110 primary hepatic carcinoma(PHC).Formalin-fixed and paraffin-embedded seetions were stained by ABC method forHBxAg,and by PAP method for HRsAg and HBcAgOf the 110 cases of chronic hepatitis,72(65.5％)were positive HBxAg in the liver cells,66(60％)were postitive in HBsAg and 35(31.8％)in HBcAg.Among the 108 eases of drrhosis,84(77.8％)revealed to be HBxAg positive in the liver cells,73(67.6％)were demonstrated to beHBsAg-positive and 18(16.7％)were shown to be HBcAg-positive.Among the 110 eases of pri-mary hepatic carcinoma,64(58.2％)showed HBxAg-positive reaction in cancerous tissues.Therates of positive HRsAg and HBcAg in tumor tissues were 15.5％ and 10.9％,respectively.Six-ty-three(78.8％)of 80 cases of the non-cancerous hepatic tissues displayed HBxAg positivenessand the rates of positive HRsAg and HBcAg in the non-tumor tissues were 47(58.8％)and 21(2.6.3％),respectively.The above-mentioned results sugared that the detection rote of HBxAg inchronic hepatitis,cirrhosis and PHC was higher than that of HBsAg and HBcAg.This studydemonstrates a dose relationship between chronic hepatitis,cirrhosis,PHC and chronic persistentinfection of hepatitis B virus(HBV).Persistent chronic HBV infection plays an important role inthe pathogenesis of chronic hepatitis, cirrhosis and PHC.It is possible that the detection ofHBxAg with anti-HBx could be an additional new diagnostic marker for HBV infection.Howev-er,the role of HBxAg in the pathogenesis of chronic liver diseases needs to be furtherinvestigated.     

丙型肝炎病毒核心抗原检测的临床应用探讨

[目的]探讨HCV核心抗原在HCV感染诊断和治疗中的作用。[方法]对173例HCV感染患者血清和82例健康对照者血清,使用ELISA法分别检测抗-HCV、总的HCV核心抗原,使用荧光定量RT-PCR检测HCV RNA。[结果]173例HCV感染者抗-HCV均为阳性。HCV RNA阳性率为49.7%(86/173);HCV核心抗原阳性率为36.4%(63/173)。二者差异有统计学意义(P<0.05)。86例HCV RNA阳性标本中,HCV核心抗原阳性60例(69.8%);HCV核心抗原阴性26例(30.2%)。82例健康对照者抗-HCV、HCV核心抗原、HCV RNA均为阴性。[结论]在没有HCV RNA检测资格的实验室,HCV核心抗原检测可作为病毒存在和复制的一个指标,作为抗-HCV检测的补充试验。     

丙肝病毒核心抗原检测在丙肝病毒感染诊断中的作用研究

目的探讨丙肝核心抗原检测在丙肝感染诊断中的作用。方法收集175例丙肝抗体检测标本同时检测HCVAb、HCV游离抗原、HCV总抗原、HCVRNA。结果 75例HCVAb阳性反应标本,检出HCV游离抗原阳性17例,HCV总抗原阳性54例,HCVRNA阳性50例;100例HCVAb阴性反应标本,检出HCV游离抗原阳性1例,HCV总抗原阳性2例,HCVRNA阳性1例。结论 HCV核心抗原是HCV早期感染的标志之一,检测HCV核心抗原有利于HCV感染的早期诊断。     

Clinical application and analysis of hepatitis C virus NS3 antigen detection by ELISA in human serum

Background Hepatitis C virus （HCV） core antigen assays have been produced to exclude infectious donations collected during the preseroconversion window phase （PWP）. For the same purpose, we evaluated the specificity and sensitivity of a novel hepatitis C virus NS3 antigen detection immunoassay and the application of this assay in clinical diagnosis. Methods Samples from 77 healthy subjects, 173 anti-HCV positive patients and 3708 hepatitis patients other than HCV positive were tested with the HCV NS3 antigen assay. Some HCV NS3 antigen positive samples were further validated with HCV-RNA, neutralization and immunodot assays. Twenty-five sequential samples from 11 HCV NS3 antigen positive patients were subjected to kinetic study. Results Only 48 （1.3%） of 3708 anti-HCV negative samples were positive for HCV NS3 antigen. Among them, 44 of 3030 samples from patients only infected with HBV were HCV NS3 antigen positive, 4 of the 445 samples from patients infected with other type hepatitis were HCV NS3 antigen positive. In addition, 42 （24.3%） of 173 anti-HCV positive samples were HCV NS3 antigen positive and all 77 samples from healthy subjects were negative to HCV NS3 antigen assay. Of the 15 HCV NS3 antigen positive samples, 9 （60%） were HCV-RNA positive. The neutralization and positive percentage of immunodot assay for 23 HCV NS3 antigen positive sera were 87.0% （20/23） and 69.6% （16/23） respectively. Of the 25 sequential samples from 11 HCV NS3 antigen positive patients, there was a negative correlation between the OD values and the duration of test （r=-0.989, P〈0.05）, and there were correlations among their HCV NS3 antigen, HCV-RNA and anti-HCV Utres. The anti-HCV antibodies of two sera were detected while their OD values of HCV NS3 antigen decreased gradually. Conclusions The HCV NS3 antigen detection assay showed perfect specificity and high sensitivity. Thus, it would be useful and economical as a routine test in laboratories for early diagnosis of HCV infection and prevention.     

丙型肝炎病毒核心抗原的检测及其临床意义

目的 探讨HCV 核心抗原在HCV 感染诊断和治疗中的作用.方法 丙肝核心总抗原检测采用酶联免疫法;丙型肝炎抗体检测采用第三代酶联免疫法.所有血清标本均进行HCV-RNA 和肝功能检测.丙肝核酸定量采用PCR- 荧光探针法;肝功能检测采用紫外连读检测法.结果 90 例中,HCV-cAg 检测阳性率为42.22%(38/90).HCV-cAg 阳性组HCV-RNA 阳性率100%(38/38),显著高于HCV-cAg 阴性组的71.15%(37/52).阳性组丙氨酸氨基转移酶(alanine aminotransferase,ALT)异常率63.16%(24/38),显著高于HCV-cAg 阴性组的40.38%(21/52),两组间比较(P<0.05).HCV-cAg 阴性组中59.62% (31/52)的患者ALT 在正常范围内(ALT<40U/L),显著高于HCV-cAg 阳性组的36.84%(14/38) ;而HCV-cAg 阳性组中36.84%(14/38)ALT 值均>100U/L,明显高于HCV-cAg 阴性组的9.62%(5/52).两组间比较(P<0.05).丙肝核心抗原HCV-cAg 和HCVRNA有很好的正相关性,HCV-cAg 与ALT 水平也有一定相关性,可以反映肝功损害程度.结论 丙肝核心抗原HCV-cAg是慢性丙肝感染良好的监测指标.     

HCV核心抗原C33肽单克隆抗体的研制与初步鉴定

用ＨＣＶ核心抗原（Ｃ３３肽）与鼠血清白蛋白交联后免疫ＢＡＬＢ／ｃ小鼠，用杂交瘤技术获得有实用价值的二株ＭｃＡｂ。此二株ＭｃＡｂ除与免疫抗原有较强的抗原－抗体反应外，与ＨＣＶ核心抗原ＣＰ１０也有很好的反应；但与ＨＣＶＮＳ３、ＶＳ５及核心区ＣＰ９抗原无反应性。在竞争ＥＬＩＳＡ中对抗ＨＣＶ－ＩｇＧ阳性血清有较好的抑制作用。     

检测丙型肝炎核心抗体的合成肽酶联免疫试验

为建立一种以酶联免疫试验检测丙型肝炎病毒的核心抗体,按病毒氨基酸序列C区合成一个36-寡肽,将其作为试剂抗原包被固相,以捅获待检血清的相应抗体,以酶标抗入IgG将其检出。输血后非甲非乙肝炎32例检出抗体20例(62.5％),其中发病1个月内的13例中检出8例。特异性由抑制试验确定。17例阳性血清可用聚合酶链反应检出病毒。结果表明,完善这一系统可能发展为试剂盒。     

原发性肝癌患者乙型肝炎病毒与丙型肝炎病毒感染标志物的分析

了解原发性肝癌患者乙型肝炎病毒和丙型肝炎病毒的感染状况,并探讨原发性肝癌与乙型肝炎病毒和丙型肝炎病毒的关系。用酶联免疫吸附法对８２例肝癌患者、８０例肿瘤对照组和１０３例健康对照组进行了乙型肝炎病毒和丙型肝炎病毒感染标志物的检测。     

保定市丙型肝炎病毒基因型分布特征及临床意义

目的 了解保定市丙型肝炎病毒（HCV）基因型分布特征及其与丙型肝炎病毒核糖核酸（HCV-RNA）载量、肝病程度的相关性,为其防治提供依据。方法 对2010年1月—2017年9月758例HCV-RNA阳性者进行HCV的基因型检测,分析其基因型与HCV-RNA含量和肝病程度的相关性。结果 758例HCV感染患者中1b型和2a型分别为50.79%（385/758）和36.68%（278/758）。HCV基因型的分布特征与性别、感染途径及种族差异均无统计学意义（P>0.05）。1b型患者ALT、AST、GGT及HCV-RNA水平均高于2a型和其他型患者,差异有统计学意义（P<0.05）。不同程度肝病患者的HCV基因型分布差异有统计学意义（P<0.05）,1b型丙肝肝硬化及肝癌比例高于其他基因型,差异有统计学意义（P<0.05）。1b型患者对长效干扰素加利巴韦林治疗的ETVR应答率（57.92％ vs 83.09％）和SVR应答率（24.68％ vs 60.79％）均明显低于2a型患者（P<0.05）。相关分析显示,1b型患者ALT与HCV-RNA呈正相关（r=0.685,P<0.01）。结论 保定市丙型肝炎病毒基因型流行株为1b型和2a型,其中1b型与丙肝肝硬化、肝癌的产生有一定的相关性。     

人原发性肝细胞癌中HBxAg的免疫组化及免疫电镜研究

作者用免疫组化和胶体金免疫电镜方法,观察乙型肝炎x抗原(HBxAg)在肝癌组织(35例)及癌旁肝组织(29例)内的分布及亚细胞定位。结果显示,HBxAg在癌组织及癌旁肝组织内的检出率分别为60.0％和82.8％,阳性物质主要位于细胞浆内。电镜观察,HBxAg主要定位于粗面内质网和胞浆基质中,少数位于核内,并可见游离核糖体及核孔附近阳性,推测HBxAg在内质网及游离核糖体上合成,并可经核孔进入核内。     

丙型肝炎患者外周血单个核细胞HCV和Fas抗原检测

目的：探讨丙型肝炎（丙型）患者外周血单个核细胞（ＰＢＭＣｓ）内丙型肝炎病毒抗原（ＨＣＶＡｇ）及Ｆａｓ抗原（ＦａｓＡｇ）表达状况及意义。方法：采用ＨＣＶ核心区及ＮＳ３区单克隆抗体,对２８例慢性丙肝患者ＰＢＭＣｓ进行免疫组化检测,并同步检测ＦａｓＡｇ。结果：ＨＣＶＡｇ及ＦａｓＡｇ阳性率分别为８５．７％（２４／２８）和８２．２（２３／２８）,经相关分析显示。两者表达呈显著正相关（ｒ＝０．５４６,Ｐ〈０．     

时间分辨免疫荧光分析检测乙型肝炎病毒表面抗原

目的 时间分辨荧光免疫分析技术(TRFIA)是当前标记免疫分析研究应用领域的热点之一,具有高灵敏性和高特异性,用于生物分子的超微量检测.本研究者在建立一种灵敏度高、特异性强、操作简便的检测方法.方法 以Eu-DTTA{N1-(P-异硫氰基苄基)-二乙三胺四乙酸铕钠}为示踪物,建立了用TRFIA检测乙型肝炎血清学标志物乙型肝炎病毒表面抗原的方法.结果 所建立的标准曲线分析范围分别为0.2～300 ng/ml,分析灵敏度为0.05 ng/ml,检测参考标准品的批内变异系数均小于10%,与免疫放射测定同时定量检测多份血清样本,相关系数高达95%.结论 TRFIA分析范围宽,灵敏度高,操作简便,具有优越的定量检测能力,价格适中,十分适合临床推广应用.     

戊型肝炎患者肝组织中HEV抗原的检测研究

目的：用抗－ＨＥＶ ＯＲＦ２单克隆抗体和抗－ＨＥＶ ＯＲＦ２单链可变区抗体研究急性戊型肝炎的免疫病理学,探讨二者在临床病理诊断中的应用价值。方法：（１）以杂交瘤技术制备的抗－ＨＥＶ单克隆抗体（ＭｃＡｂ,ｍｏｎｏｃｌｏｎａｌ ａｎｔｉｂｏｄｙ）作为第一抗体,用免疫组化ＬＳＡＢ法检测急性戊型肝炎患者肝组织中的ＨＥＶ抗原。（２）用抗－ＨＥＶ ＯＲＦ２单链可变区抗体以间接酶标法检测肝组织内的ＨＥＶ抗原。结果：用抗－ＨＥＶ单克隆抗体从１４例急性戊型肝炎肝标本中检出９例ＨＥＶ抗原阳性者（６４．２８％）,其中８例阳性标本用抗－ＨＥＶ ＯＲＦ２单链可变区抗体检测ＨＥＶＡｇ均为阳性。ＨＥＶ抗原在肝细胞浆表达以胞浆弥漫型为主,尚可见膜型表达。ＨＥＶ抗原阳性细胞集中分布有病变明显区,伴淋巴细胞浸润,病变较轻区也可见随机散在分布,部分     

Allele polymorphisms of interleukin-lO and hepatitis B,C virus infection

Background Interleukin 10 （IL-10） is an important cytokine with anti-inflammatory, anti-immune and anti-fibrotic functions. This study aimed at evaluating the relationship between allele polymorphisms in the IL-10 promoter region and hepatitis B virus （HBV） or hepatitis C virus （HCV） infection. Methods The odds ratios （ORs） of IL-10 allele distributions in patients with HBV or HCV infection were analyzed against healthy controls. All the relevant studies in PubMed were identified, and poor qualified studies were excluded. The meta-analysis software REVMAN 4.2 was applied for investigating heterogeneity among individual studies and summarizing all the studies. The publication bias was also evaluated. Results This study demonstrated a significant association between the IL-10-592 A/C polymorphism and HBV infection in the Asian population under the overall effect size of allele A versus C. In our subgroup meta-analysis, we found a significant association of IL-10-592 A/C polymorphism to HCV infection susceptibility in Asian populations, although sensitivity analysis showed that the combined result was not associated with the worldwide population. Other IL-10 allele polymorphisms were not associated with HBV or HCV infection. Conclusion IL-10-592 A/C allele might be a risk factor for HBV or HCV in Asians but not in Europeans.     

丙型肝炎病毒C33抗原在肝组织中的表达

目的：观察ＨＣＶＣ３３抗原在慢性丙型肝炎患者肝组织中的分布。方法：４例慢性丙型肝炎患者肝组织用免疫组化法作ＨＶＣＣ３３标记。结果：ＨＣＶ３３抗原不仅分布于肝细胞核，胞浆及肝细胞膜中，而且分布肝窦内皮细胞及肝内脉巴细胞的胞浆及胞膜上。结论：ＨＣＶ抗原存在于上述肝细胞内，是慢性丙型肝炎的重要标志。     

Fas和FasL在丙型肝炎肝组织中的表达及意义

目的了解丙型肝炎患者肝细胞损伤与Fas抗原表达的关系。方法采用免疾组织化学方法显示Fas抗原与Fas配体（FasL）在丙型肝炎肝组织的分布，并同时运用原位杂交方法，检测肝组织中的丙型肝炎病毒（HCV）RNA。结果Fas抗原主要位于肝细胞胞浆。Fas抗原表达阳性细胞在肝小叶中多呈散在或灶状分布，汇管区周围和碎屑样坏死区内阳性表达较强。FasL大多位于肝内浸润的淋巴细胞，某些病例肝细胞胞浆也可呈弥漫强阳性。Fas抗原表达的阳性率与肝组织炎症程度相关。结论在丙型肝炎患者，Fas抗原表达水平与炎症活动度有关，未发现（HCV）RNA原位杂交阳性细胞与表达Fas抗原的肝细胞之间的对应关系。     

In vitro expression of hepatitis C virus non-structure 5 antigen in the HepG2 cell line

Summary To establish a cell line as a model system for HCV infection and propagationin vitro, a human HepG2 cell line was incubated with a HCV RNA positive serum. The sABC immunological techniques and gold-labeled colloid electron microscopy method were employed to examine the viral proteins in those cells. The HCV non-structure 5 antigen was first detected in the HepG2 cells 72 h after incubation. The antigen was continuously observed in the cytoplasm as well on the membrane of the HepG2 cells even after 1, 2, 3 and 4 weeks after incubation. The observation of HCV non-structure 5 antigen continuously expressed in the HepG2 cells strongly indicates that the cells may have been infected by HCV virus. Therefore, the HepG2 cell line may serve as a potential host for establishment of HCV infection and propagationin vitro. This project was supported by a grant of the National Educational Committee for return scholars (No.)