肝癌患者HBxAg与Fas/FasL表达的研究

目的：探讨肝癌患者肝组织中 ＨＢｘＡｇ与 Ｆａｓ／ＦａｓＬ的表达状况及血清中 ＨＢＶ Ｘ基因与 Ｆａｓ／ＦａｓＬ的水平。方法：用ＳＰ法对３４例肝癌组织ＨＢｘＡｇ、Ｆａｓ与ＦａｓＬ表达状况进行分析,用ＥＬＩＳＡ法对３０例肝癌、３２例肝硬化及２０例正常人血清ｓＦａｓ与 ｓＦａｓＬ含量进行分析,并用ＰＣＲ法对肝癌与肝硬化患者血清ＨＢＶ Ｘ基因表达进行检测。结果：ＨＢｘＡｇ、Ｆａｓ与ＦａｓＬ在肝癌表达的阳性率分别为９７．０６％、８５．２９％及１００％。阳性信号均主要位于胞浆,且三者可在同一癌组织的相同区域出现阳性染色,Ｒｅｄｉｔ分析示三者的阳性程度无统计学差异（Ｐ＞０．０５）。血清ｓＦａｓ在肝癌、肝硬化与正常人中的含量分别为（７２２．９７±３２１．１３）μｇ／Ｌ、（８０１．９０±４１９．９４）μｇ／Ｌ和（２２４．０７±１４８．２３）μｇ／Ｌ,其中肝癌与肝硬化组显著高于正常人（Ｐ＜ ０． ０１）;血清 ｓＦａｓＬ在肝癌、肝硬化与正常人中的含量分别为（１５２． ２７±７． ９９）μｇ／Ｌ、（ １６２． ９７±１２． ４０）μｇ／Ｌ和（１５４．９９± ６．９６）μｇ／Ｌ,其中肝硬化组含量高于肝癌与正常人（ Ｐ＜０．０１）。血清ＨＢＶ　Ｘ基     

原发性肝癌丙肝病毒基因及其抗体的检测

本文报道运用巢式ＰＣＲ，ＥＬＩＳＡ等方法检测我国原发性肝癌，癌旁组织和外周血中ＨＣＶ－ＲＮＡ抗－ＨＣＶ，同时监测了ＨＢｓＡｇ。所用引物位于ＨＣＶ基因５＇－端非编码区域。结果７６例肝癌病人血清中抗－ＨＣＶ阳性９．２％（７／７６），ＨＣＶ－ＲＮＡ阳性１３．２％（１０／７６）；３４例肝癌组织中ＨＣＶ－ＲＮＡ阳性１１．８％（４／３４），１例抗ＨＣＶ阴性的病人在癌及癌旁组织中检测到ＨＣＶ－ＲＮＡ，提示我国Ｈ     

人原发性肝细胞癌组织中Fas,FasL表型检测的意义

探讨人原发性肝细胞（ＨＣＣ）组织中Ｆａｓ、Ｆａｓ配体（ｆａｓｌｉｇａｎｄ,ＦａｓＬ）的表达水平与ＨＣＣ发生和预后的关系。方法采用免疫组化（ＳＰ）法,检测３１例ＨＣＣ原发灶中Ｆａｓ、ＦａｓＬ的表达水平。结果３１例ＨＣＣ组织中有２３例（７４．２％）ＦａｓＬ表达阳性,明显高于癌旁组织,Ｐ〈０．０５,Ｆａｓ阳性表达１６例（５１．６％）与癌移和无转移的ＨＣＣ患者中,ＦａｓＬ表达和阳性率分别为８５．７％和５０     

丙型肝炎病毒核心抗原及HBxAg在肝硬化、肝细胞肝癌中分布的免疫组化研究

研究丙型肝炎病毒核心抗原在肝硬变、肝细胞肝癌中的分布规律及意义。方法以免疫组织化学方法检测丙型肝炎病毒（ｈｅｐａｔｉｔｉｓＣｖｉｒｕｓ，ＨＣＶ）核心抗原及ＨＢｘＡｇ在肝硬化、肝细胞癌及其癌旁肝组织中的定位及分布。结果ＨＣＶ核心抗原既定位于肝细胞或癌细胞胞核中，又可定位于这些细胞的胞浆中。在不同的病例，有时以胞浆阳性为主，有时以胞核阳性为主，或二者同时存在。肝硬化组织中，ＨＣＶ核心抗原胞浆阳性细胞多呈灶性分布，而核阳性病例阳性细胞则为弥漫分布；肝细胞肝癌中ＨＣＶ核心抗原以弥漫胞核阳性多见，癌旁肝组织多为ＨＣＶ核心抗原胞浆阳性。ＨＣＶ核心抗原在肝硬化、肝细胞肝癌及癌旁肝中的阳性率分别为６７．３％（６６／９８），７５．０％（７８／１０４）及４８．１％（２５／５２），χ２检验，肝细胞癌中核心抗原的胞核阳性率明显高于其在肝硬化及癌旁肝中的胞核阳性率（Ｐ＜０．０１）。核心抗原在肝细胞肝癌中，其细胞核阳性率，明显高于细胞浆阳性率（Ｐ＜０．０１）。结论ＨＣＶ感染在我国肝硬化、肝细胞肝癌中比较普遍，除ＨＢＶ以外，ＨＣＶ可能在我国肝硬化、肝细胞癌的发生中起着重要作用     

肝癌,肝硬变患者的戊型肝炎病毒感染

目的研究肝癌和肝硬变患者的肝炎病毒感染情况。方法用酶联免疫吸附测定（ＥＬＩＳＡ）法测定患者血清中的乙型肝炎病毒表面抗原（ＨＢｓＡｇ）、丙型肝炎病毒抗体（抗ＨＣＶ）和戊型肝炎病毒抗体（抗ＨＥＶ）。结果肝癌患者中抗ＨＥＶ阳性率为５８．９％（６３／１０７），ＨＢｓＡｇ阳性率为６９．２％（７４／１０７），抗ＨＣＶ阳性率为１０．３％（１１／１０７），肝硬变患者的阳性率依次为６３．０％（１７／２７）、７４．１％（２０／２７）、７．４％（２／２７）。只有抗ＨＥＶ阳性而ＨＢｓＡｇ和抗ＨＣＶ阴性的肝癌患者有１３例（１２．２％）。仅ＨＢｓＡｇ阳性而抗ＨＥＶ和抗ＨＣＶ阴性的有２４例（２２．４％），仅抗ＨＣＶ阳性而抗ＨＥＶ和ＨＢｓＡｇ阴性的有３例（２．８％）。全部阴性的有１０例（９．４％）。肝硬变患者中仅抗ＨＥＶ阳性而抗ＨＣＶ和ＨＢｓＡｇ阴性的有５例（１８．５％），仅ＨＢｓＡｇ阳性而抗ＨＥＶ和抗ＨＣＶ阴性的有９例（３３．３％），全部阴性的有１例（３．７％）。结论除ＨＢＶ和ＨＣＶ外，ＨＥＶ感染似乎在肝癌变及肝硬变中也起着一定的作用     

肝癌及癌旁组织中HBsAg表达的研究

应用免疫组织化学技术，检测４０例肝癌（ＨＣＣ）及癌旁组织中ＨＢｓＡｇ表达，阳性率达８２．５％，表明ＨＢＶ感染与ＨＣＣ发生密切相关。ＨＢｓＡｇ在ＨＣＣ中表达较癌旁组织中少而弱，可能系肝细胞癌变后不利于ＨＢｓＡｇ表达及Ｓ基因发生重排或缺失之故。碎点状ＨＢｓＡｇ小包涵体，其可能为ＨＣＣ中ＨＢｓＡｇ的特殊表现形式、小细胞ＬＣＤ中ＨＢｓＡｇ表达显著高于癌旁其他类型病变，支持其更接近癌前病变的观点。ＨＢｓＡｇ染色可作为鉴别ＨＲＶ感染肝病中癌前病变与早期癌及高分化癌的一种辅助手段。     

广西肝细胞癌与丙型肝炎病毒的相关性

目的研究广西肝癌高、中、低危区的癌及癌旁肝组织中丙型肝炎病毒（ＨＣＶ）的表达,探讨ＨＣＶ在广西肝细胞癌（ＨＣＣ）病因中的地位。方法用ＨＣＶ的Ｃ区、ＮＳ３和ＮＳ４区３种单克隆抗体及抗ＨＢｓＡｇ抗体进行检测。免疫组化染色用ＳＰ法。结果ＨＣＣ高、中、低危区肝癌中ＨＣＶ阳性率依次为３８．１％（２４／６３）、３７．１％（２３／６２）和３９．０％（３０／７７）;ＨＢｓＡｇ阳性率依次为８４．１％、８３．８％和８４．４％。ＨＣＣ中ＨＣＶ感染的总数为７７／２０２（３８．１％）,乙型肝炎病毒感染总数为１７０／２０２（８４．２％）。结论ＨＣＣ经常伴随乙型肝炎病毒（ＨＢＶ）感染,但ＨＣＶ可见于１／３的ＨＣＣ。在广西肝癌高、中、低发地区,ＨＢＶ与ＨＣＶ感染情况无明显差别。     

抗HBx单抗应用于肝癌导向治疗的实验研究

本研究应用基因工程技术制备的１７ＫＤ乙型肝炎病毒（ＨＢＶ）Ｘ蛋白（ＨＢｘＡｇ）免疫动物，成功地制备了抗－ＨＢｘ单克隆抗体，以此抗体对原发性肝癌（ＨＣＣ）组织行相应抗原检测，并同时进行乙型肝炎表面抗原（ＨＢｓＡｇ）及乙型肝炎病毒核心抗原（ＨＢｃＡｇ）的检测。在ＨＣＣ标本中，ＨＢｘＡｇ在癌内及癌周肝组织中的阳性率分别为５６％及４８％，ＨＢｓＡｇ为１６％，及７４％。ＨＢｃＡｇ仅见于癌周，阳性率仅为１８％。结果显示：ＨＢｘＡｇ在ＨＣＣ标本中的表达是ＨＢＶ标志物中最活跃的。在此基础上还进行了１３１Ｉ－ＨＢｘ单抗在荷人肝癌裸鼠体内的放射分布研究，给药后第７天瘤／血比值为１．５，瘤／肝比值为４．５，而对照组为０．４４及１．０４，表明１３１Ｉ－ＨＢｘ单抗对肝癌组织有较强的特异性亲和力，有可能成为新的载体用于肝癌的导向治疗     

人原发性肝内管细胞癌及其癌旁肝组织内乙型肝炎病毒基因的表…

为了从分子水平探讨乙型肝炎病毒与肝内胆管细胞癌的发病关系，应用原位分子杂交技术，在４０例肝内胆管细胞癌及其癌旁肝细胞内检测ＨＢＶ ＤＮＡ，ＨＢＶ的ｐｒｅ－Ｓ基因，Ｓ基因，Ｘ基因和Ｃ基因。结果显示，在４０例中，３３例ＨＢＶ ＤＮＡ呈阳性信号占８２．５％；Ｘ基因阳性为３１例，占７７．５％；ｐｒｅ－Ｓ基因阳性２６例，占６５．０％，Ｓ基因阳性２４例，占６０．０％；Ｃ基因阳性２７例，占６７．５％。在后４种基     

乙型肝炎病毒X基因与肝癌p53突变蛋白的表达

应用免疫组织化学（ＡＢＣ法）对２３例肝癌和癌旁组织中ｐ５３突变蛋白进行检测，同时对其中ＨＢＶＸｍＲＮＡ进行原位杂交（ＤＮＡ－ｍＲＮＡ）检测。结果发现，９例肝癌及其癌旁组织中ｐ５３突变蛋白和ＨＢＶＸｍＲＮＡ均为阳性，１１例均为阴性，另３例仅ＨＢＶＸｍＲＮＡ阳性，χ２检验显示ｐ５３突变蛋白与ＨＢＶＸｍＲＮＡ之间有明显相关关系（Ｐ＝０．０００２７），提示ｐ５３基因突变与ＨＢＶＸ基因整合表达有关，ｐ５３基因突变可能是ＨＢＶ致肝癌作用机制之一。     

HCV核心抗原、NS3抗原在肝癌及癌旁肝组织的表达及意义

 采用SP法（链菌素抗生物素蛋白—过氧化酶连结法）检测46例肝细胞癌及其38例癌旁肝组织中HCV核心抗原、NS3抗原和HBsAg。结果在癌组织中的检出率分别为21．7％、23．9％和19．6％，癌旁组织中的检出率分别36．8％、39．5％和78．9％，核心抗原、NS3抗原伴HBsAg者分别为23.9％和19．6％，不伴HBsAg者分别为10．9％和13％；HBsAg不伴HCV核心抗原、NS3抗原者分别为47．8％和50％。两种HCV抗原均定位于肝细胞和肝癌细胞的胞浆内，在癌旁组织中均有围核分布的特点，其阳性染色细胞呈弥漫、灶状和散在分布。本研究结果提示HCV核心抗原、NS3抗原的检测有可能作为HCV感染的新标志物；HCV和HBV与肝细胞癌的发生可能密切相关，部分患者与HBV和HCV的双重感染可能有关；HCV核心抗原、NS3抗原在肝癌发生中的真正作用，尚待研究。     

Rate of incidence of hepatocellular carcinoma in patients with compensated viral cirrhosis.

BACKGROUND: Cirrhosis of viral etiology due to hepatitis B virus (HBV) or hepatitis C virus (HCV) infection is a risk factor for hepatocellular carcinoma (HCC). The current study evaluated the rate of incidence of HCC in patients with compensated cirrhosis of viral etiology. METHODS: Two hundred fifty-nine cirrhotic patients (66 hepatitis B surface antigen [HBsAg] positive, 166 HCV positive, and 27 HBsAg/HCV positive) were longitudinally examined every 6 months by serum alpha-fetoprotein test and liver ultrasonography. The rates of incidence of HCC were calculated by the person-years method. The Kaplan-Meier method was used to estimate the cumulative probability of HCC development. Differences in survival time were evaluated by a log rank test. Independent predictors of HCC development were estimated by Cox proportional hazard regression analysis. RESULTS: During a mean follow-up of 64.5 months, HCC developed in 51 (19.7%) patients: in 34 of 166 HCV positive subjects (20.5%) (mean follow-up, 66.3 months), in 6 of 66 of those HBsAg positive (9.1%) (mean follow-up, 55.06 months), and in 11 of 27 of those with dual HBsAg/HCV infection (40.7%) (mean follow-up, 76.4 months). The rate of incidence of HCC per 100 person-years of follow-up was 3.7 in HCV positive subjects, 2.0 in those HBsAg positive, and 6.4 in those with dual infection. Cumulative HCC appearance rates in HBsAg positive, HCV positive, and HBsAg/HCV positive subgroups were 10%, 21%, and 23% at 5 years, 16%, 28%, and 45% at 10 years, and 16%, 40%, and 55% at 13 years, respectively. Multivariate analysis indicated that age >50 years (hazard risk [HR], 4.5; 95% confidence interval [CI] = 2.1-9.4), male gender (HR, 2.8; 95% CI = 1.1-5.3), and HBsAg/HCV coinfection (HR, 2.3; 95% CI = 1.1-4.6) were independent predictors of HCC development. CONCLUSIONS: These findings confirm that male gender and more advanced age (>50 years) are risk factors for HCC in patients with cirrhosis. Furthermore, the data indicate that subjects with dual HBsAg/HCV infection are at highest risk for HCC. Surveillance programs for early detection of HCC should focus especially on these patients.     

HCV感染对肝癌一些癌相关基因产物表达的影响

目的 研究丙型肝炎病毒（ＨＣＶ）的致癌机理。方法 以免疫组化ＡＢＣ法对４６例肝细胞癌（ＨＣＣ）及其癌旁肝组织的ＨＣＶＣＰ１０、ＮＳ３及ＮＳ５抗原和rasp２１、c-myc、c-erbB-2、突变型p５３及p１６蛋白进行染色,并将ＨＣＣ分为ＨＣＶ抗原阳性组和ＨＣＶ抗原阴性组,对比分析ＨＣＶ感染对上述癌相关基因蛋白产物表达的影响。结果 ４６例ＨＣＣ患者中,１～３种ＨＣＶ抗原性２０例（４３．５％）,２～３种ＨＣＶ抗原阳性     

多发性骨髓瘤与HBV、HCV及EBV病毒感染的关系

目的:探讨多发性骨髓瘤（multiple myeloma,MM）发病与乙肝病毒（HBV）、丙肝病毒（HCV）和EB病毒（EBV）感染之间的相关性。方法:采用多中心、以医院为基础的1∶1配对病例对照研究。265例MM病例来自西安市五家三级甲等医院血液科在2011年10月至2014年9月期间住院患者。对照来自同期、同院住院的非肿瘤患者,与病例在性别和年龄（±5岁）上匹配。ELISA方法检测研究对象血清中的HBsAg、抗-HBc、抗-HCV和抗-EBV。应用Logistic回归分析不同病毒感染与MM发病的关联性。结果:MM组HBsAg阳性率与对照组相比差异无显著性意义（7.55% vs 4.49%,P>0.05）。MM组抗-HBc阳性率明显高于对照组（45.82% vs 33.64%,P<0.01）,OR值为1.66（95%CI:1.17~2.36）。将HBsAg和抗-HBc合并分析发现两组差异有统计学意义（P<0.01）,OR值1.82（1.28~2.58）。HCV和EBV病毒阳性率在两组之间的差异无统计学意义（P>0.05）。结论:HBV感染增加MM的发病风险,而HCV和EBV感染与MM发病风险无明显关联。     

肝外胆管癌病人中的丙型肝炎病毒和乙型肝炎病毒感染

Objective:In China, the incidence of extrahepatic bile duct carcinoma (EBDC) tends to increase over the past decades. The etiology of the noted increase in EBDC is not identified. Approximately, in a half of the overall Chinese patients with EBDC, the causative factors in the development of EBDC have not been demonstrated. There is a high prevalence of hepatitis C virus (HCV) or hepatitis B virus (HBV)in China, both of which can induce malignant transformation of infected cells and strongly associated with hepatocellular carcinoma (HCC).In this study,EBDC tissues from Chinese patients were examined for the presence of HCV and HBV infection to investigate further the potential causes of EBDC. Methods:HCV NS5 protein and HBsAg were detected by labeled streptavidin biotin (LSAB) method; HCV RNA and HBV DNA were detected by in situ polymerase chain reaction (IS-PCR) in formalin fixed, paraffin embedded specimens from 51 Chinese patients with EBDC. HCV RNA and HBV DNA were detected by IS-PCR in 34 Chinese patients with specimens of benign lesions of hepatobiliary tract(control group). Results:In 51 case tissue sections of EBDC, NS5 protein was detected in 14 (27.5%), and HBsAg in 5 (9.8%), HCV RNA in 18(35.4%) and HBV DNA in 8 (15.9%), respectively, of which HCV and HBV co-infection was detected in 2 (3.9%). In 34 case tissue sections of the control group, HCV RNA was detected in 2 (5.9%), and HBV DNA in 3 (8.8%).Conclusion:In this study using standard histochemical and PCR techniques,HCV and HBV and HBV presence in EBDC tissues than would be expected on serologic grounds.The detectable rate of HCV RNA in EBDC tissues was significantly higher than in control group(x^2=9.808,P=0.002).As a result ,this study indicates that there is a correlation between the presence of HCV infection and EBDC,and HCV infection has possible ctiologic significance in the development of EBDCin China.While HBV DNA was detecled in EBDC tissues with the difference in the detectable rate of HBV.DNA being not significance betwwen EBDC tissues and the control group(x^2=0.853,P=0.356).Further research is necessary to determine the presence of a causal relationship between HCV/HBV infection and the development of EBDC.     

Genetic Polymorphisms of Fas/FasL Promoter Associated with Hepatitis C cirrhosis and HCC

Aim: The present study was performed to determine any associations of genetic polymorphisms of Fas/FasL promoter regions, at Fas670 and Fas1377 and FasL844, with hepatitis C cirrhosis and HCC, with a focus on severity of disease. Methods: Totals of 120 patients with cirrhosis and 101 with hepatocellular carcinoma (HCC) were enrolled. All had chronic HCV infection as indicated by positive anti-HCV antibodies and positive HCV RNA on real time PCR. One hundred healthy control subjects were also included in the study. Patients were subjected to full clinical, radiological and histopathological examinations. In addition to routine laboratory tests for liver function tests, Fas670 and Fas1377 and FasL844 genetic polymorphisms of Fas/FasL promoter regions were assessed by RFLP-PCR (restriction fragment length polymorphism with polymerase chain reaction). Results: Significant higher levels of the AG genotype in Fas670 and Fas1773 were observed in patients with cirrhosis and HCC (P=0.0001) as compared to control subjects. In addition, the CC genotype in FASL844 was also more common in patients (P=0.01). Furtehrmore, there was a significant association of substitution of A by G alleles in Fas670 and Fas1773 with advanced BCA staging (P=0.02, P=0.0001 respectively) and larger tumor size >5cm (P=0.01, P=0.0001 respectively) and in Fas670 with advanced pathological grading (P=0.0001). Moreover the CC genotype of FASL844 was significantly linked with advanced BCA, large tumor size >5cm and advanced pathological grading (P=0.0001). Conclusion: The findings of the present study highlight associations of genetic polymorphisms of promoter regions in Fas and Fas L with cirrhosis and HCC associated with chronic HCV. Support was also obtained for the conclusion that single nucleotide polymorphisms of the Fas/ FasL system impact on clinical and histopathological grading of HCCs. Further large scale studies are recommended for confirmation.     

Prospective study on the risk of hepatocellular carcinoma among hepatitis C virus-positive blood donors focusing on demographic factors, alanine aminotransferase level at donation and interaction with hepatitis B virus

The risk for hepatocellular carcinoma (HCC) among asymptomatic hepatitis C virus (HCV) carriers is not well understood. A community-based prospective study was conducted for over 8 years by record linkage to the Osaka Cancer Registry. The subjects were 1,927 individuals who were positive for anti-HCV through screening for second-generation HCV antibody (passive hemagglutination assay: >or= 2(12)) in voluntary blood donation. The risk factors for HCC and interaction between HCV and hepatitis B virus (HBV) infection were evaluated by including additional blood donors: 2,519 individuals positive for hepatitis B virus surface antigen (HBsAg) alone, 25 positive for both anti-HCV and HBsAg, 150,379 negative for both anti-HCV and HBsAg. The incidence of HCC (/10(5) person-years) among the HCV-positive individuals increased with age in both genders, ranging from 68 to 1,306 among those aged 45-74 years. In the HCV-positive individuals, the cumulative risk of developing HCC between the ages of 40 and 74 year was 21.6% among males and 8.7% among females. A stepwise increase in risk was noted as the serum alanine aminotransferase level increased or serum cholesterol level at baseline decreased in multivariate Cox proportional hazard analysis. The 9-year cumulative incidence of HCC among individuals positive for HCV alone, those positive for HBsAg alone and those positive for both was 3.0%, 2.0% and 12.0%, respectively. The age-and-sex-adjusted rate ratio was 126, 102 and 572, respectively, when those negative for both were used as a reference. The results demonstrate an increased risk for HCC among asymptomatic HCV-positive individuals in Japan. Coinfection with HBV and HCV carried a superadditive risk for HCC.     

Case-Control Study of Hepatocellular Carcinoma among Koreans Living in Osaka, Japan

Mortality rates from liver cancer among Koreans living in Osaka are 2–3 times higher than those among Japanese. Our previous study revealed that chronic hepatitis B virus (HBV) infection and excessive alcohol drinking are two major risk factors for hepatocellular carcinoma (HCC) among Koreans in Osaka, although more than 70% of the HCC cases were negative for hepatitis B surface antigen (HBsAg). Using a recently developed immunoassay for detecting serum hepatitis C virus antibody (HCV-Ab), the role of HCV infection was evaluated in a case-control study. The case group consisted of 90 Korean patients who were admitted to Kyowa Hospital in Osaka, and were newly diagnosed as HCC during the period from January 1989 to December 1992. The control group consisted of 249 Korean patients admitted to Kyowa Hospital during the same period and matched in age groups to the HCC cases. Seventy-four and 16.7% of cases were positive for HCV-Ab and HBsAg, respectively. Besides, 41.1% of cases were heavy drinkers. Multiple logistic regression analysis revealed that the adjusted relative risk was 92.4 for HCV-Ab positive and 58.2 for HBsAg positive, as compared with both HCV-Ab and HBsAg negative. Elevated risk was also demonstrated for males with a history of heavy drinking. There was no significant association between the risk of HCC and a history of blood transfusion or cigarette smoking. It was concluded that chronic HCV infection plays a major role in the etiology of HCC among Koreans living in Osaka, in addition to HBV and heavy drinking.