60例肝活检组织中原位杂交及免疫组化研究

为了解在丙型肝炎病毒感染后机体组织中病毒复制与血清学改变是否协同一致及病理诊断丙型肝炎的临床意义，应用了丙型肝炎病毒（ＨＣＶ）核心区段的生物素探针，及病毒核酸的３′和５′末端非结构区（ＮＳ３和ＮＳ５）的单克隆抗体，对６０例血清学抗ＨＣＶ阴性的石蜡包埋的肝活检组织进行原位杂交及免疫组化检测，并同时检测其中乙型肝炎病毒（ＨＢＶ）核酸及ＨＢｓＡｇ、ＨＢｃＡｇ的表达。结果ＨＣＶＲＮＡ的检出率为３０％，ＮＳ３及ＮＳ５分别为４８．８％和５８．３％；ＨＢＶＤＮＡ及ＨＢｓＡｇ、ＨＢｃＡｇ检出率分别为５１．６％和４１．７％。ＨＣＶＲＮＶ的阳性信号为感染细胞浆内分布为主的棕色颗粒出现；ＨＣＶＮＳ３和ＮＳ５抗原的阳性信号亦为棕色细颗粒状，分布于感染的细胞浆中。实验结果可以说明，在丙型肝炎病毒感染的临床诊断中，组织病理学的原位杂交及免疫组化可起到血清学检测中难以取代的作用。     

乙型和丙型肝炎病毒双重感染肝组织病毒标志物的检测

为观察乙型肝炎和丙型肝炎双重感染的尸体肝组织中两种病毒核酸及抗原的分布，对１５例ＨＢＶ与ＨＣＶ双重感染的尸检肝组织用免疫组织化学法检测ＨＢｓＡｇ、ＨＢｃＡｇ和ＨＣＡｇ，Ｄｉｇｏｘｉｇｅｎｉｎ标记的探针原位杂交法分别检测ＨＢＶ ＤＮＡ和ＨＣＶ ＲＮＡ。结果：ＨＢｓＡｇ、ＨＢｃＡｇ、ＨＢＶ ＤＮＡ、ＨＣＡｇ和ＨＣＶ ＲＮＡ的阳性数分别为１２／１５（８０．０％）、１０／１５（６６．７％）、９／１５（６０     

贵州地区不同人群血清庚型肝炎病毒核酸检测和部分核苷酸序列分析

目的了解贵州地区庚型肝炎病毒感染状况,分析庚型肝炎病毒贵州株的基因特点。方法 以庚型肝炎病毒（ＨＧＶ）的ＮＳ３区两对寡核苷酸为引物,用逆转录套式聚合酶链反应（ＲＴ－ｎＰＣＲ）检测不同人血血清中庚型肝炎病毒核酸（ＨＧＶ ＲＮＡ０,并用ＰＣＲ产物直接测序分析３份阳性血清５’－非编码区的部分核苷酸序列。结果６０例正常人,３７例血液病,８６例肝病患者及１０７例静脉药瘾者血清ＨＧＶ ＲＮＡ阳性率分别为１．７     

地高辛标记探针原位杂交检测肝外组织中经血传播病毒DNA

目的 探讨血清单一经血传播病毒（ＴＴＶ）阳性患者肝外组织中ＴＴＶＤＮＡ的表达。方法 采用地高辛标记ＴＴＶＤＮＡ探针原位杂交技术检测免疫组织化学检测肝组织中丙肝病毒ＮＳ３、庚肝病毒ＮＳ５等为阴性的５例尸检肝外组织标本。结果 ３例（３／３）胰腺组织、２例（２／２）睾丸组织、１例（１／１）卵巢组织、２例（２／３）心肌组织、３例（３／５）肾组织检出阳性信号。杂交阳性信号主要为胞核型，仅在部分胰腺组织中出现     

阻止异源双链形成的定量检测GBV-C/HGV RNA的竞争性RT-PCR

为研究ＧＢ病毒Ｃ（ＧＢＶ－Ｃ）／庚型肝炎病毒（ＨＧＶ）可能的致病性及复制位点,本文作者采用竞争性逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）法检测病毒ＲＮＡ的正链及负链。共选择研究对象２３７人：第１组为６２例慢性丙型肝炎病毒（ＨＣＶ）感染者;第２组为５０例无肝炎及ＨＣＶ感染证据的患者;第３组为１２５名正常人作为对照。从血清中分离ＲＮＡ,主要采用病毒ＲＮＡ基因组的５’端非编码区（５‘ＮＣ）及非结构５（ＮＳ５）区序列作为引物。４２４及４２５引物的扩增产物（Ｐ４２４－４２５）为５’ＮＣ区核昔酸（ｎｔ）２９－３…     

血管内皮细胞生长因子在乙型肝肝炎肝组织表达的病理意义

目的 研究血管内皮细胞生长因子（ＶＥＧＦ）与乙型肝炎（乙肝）分级（Ｇ）和分期（Ｓ）的关系。方法 ２００例忆 活检肝标本用ＶＥＧＦ单克隆抗体进行免疫组化（Ｓ－Ｐ法）染色。结果 ＶＥＧＦ在肝组织表达有包浆型、膜窦圾这会面人皮细胞型;ＶＥＧＦ主要由肝细胞分泌。其次在肝窦内皮细胞、贮脂细胞、成肌纤维细胞及单型核细胞也示阳性。无明显病变的血管内皮细胞肝肝细胞ＶＥＧＦ染色阴性。提示随分级增高ＶＥＧＦ表达增强,     

固相放免法测定血清学标志物HBcAg的临床价值

目的：探讨检测乙型肝炎病毒（ＨＢＶ）血清学标志物ＨＢｃＡｇ与其它标志物的关系及其临床价值。方法：共５２６２例ＨＢＶ感染者的血清标本,应用固相放免法（ＳＰＲＩＡ）测定了乙肝表面抗原（ＨＢｃＡｇ）、乙肝表面抗体（ＨＢｓＡｂ）、乙肝ｅ抗原（ＨＢｅＡｇ）,乙肝ｅ抗体（ＨＢｅＡｂ）、乙肝核心抗原（ＨＢｃＡｇ）、乙肝核心抗体（ＨＢｃＡｂ）六项 ＨＢＶ血清学标志物。结果：发现 ＨＢｃＡｇ与 ＨＢｓＡｇ、 ＨＢｅＡｂ、 ＨＢｃＡｂ同时阳性率为 ８８．１％,明显高于其与ＨＢｓＡｇ、ＨＢｅＡｇ、ＨＢｃＡｂ同时阳性率（５５．３％）,并且在ＨＢｓＡｇ阳性、而ＨＢｅＡｇ／抗－ＨＢｅ均为阴性的感染者血清中仍有 ４０．０％检测出 ＨＢｃＡｇ。结论：ＨＢＶ血清学标志物 ＨＢｃＡｇ的检测,与其它标志物相结合,在判断 ＨＢＶ感染者血清中病毒复制活动和传染性的强弱方面,有着很重要的临床价值。     

固相放免法测定血清学标志物HBcAg的临床价值

目的：探讨检测乙型肝炎病毒（ＨＢＶ）血清学标志物ＨＢｃＡｇ与其它标志物的关系及其临床价值。方法：共５２６２例ＨＢＶ感染者的血清标本,应用固相放免法（ＳＰＲＩＡ）测定了乙肝表面抗原（ＨＢｓＡｇ）、乙肝表面抗体（ＨＢｓＡｂ）、乙肝ｅ抗原（ＨＢｅＡｂ）、乙肝核心抗原（ＨＢｃＡｇ）、乙肝核心抗体（ＨＢｃＡｂ）六项ＨＢＶ血清学标志物。结果：发现ＨＢｃＡｇ、ＨＢｃＡｂ、ＨＢｃＡｂ同时阳性率为８８．１％,明显高于其与ＨＢｓＡｇ、ＨＢｅＡｇ、ＨＢｃＡｂ同时阳性率（５５．３％）,并且在ＨＢｓＡｇ阳性、而ＨＢｅＡｇ／抗－ＨＢｅ均为阴性的感染者血清中仍有４０．０％检测出ＨＢｃＡｇ。结论：ＨＢＶ血清学标志物ＨＢｃＡｇ的检测,与其它标志物相结合,在判断ＨＢＶ感染者血清中病毒复制活动和传染性的强弱方面,有着重要的临床价值。     

80例静脉注射海因依赖者庚型肝炎病毒检测报告

目的：了解静脉注射（ｉｖ）海洛因依赖者中庚型肝炎病毒（ＨＧＶ）感染情况。方法：在ＨＧＶ５＇端非翻译区（５＇ＵＴＲ）设计引物,采用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）检测ＨＧＶ ＲＮＡ。结果：在８０例ｉｖ海洛因依赖者中,ＨＧＶ ＲＮＡ阳性率为４６．３％。结论：证实ｉｖ海洛因依赖者中ＨＧＶ感染较为常见,且吸毒时间越长,ＨＧＶ阳性率越高。     

80例静脉注射海洛因依赖者庚型肝炎病毒检测报告

目的：了解静脉注射（ｉｖ）海洛因依赖者中庚型肝炎病毒（ＨＧＶ）感染情况。方法：在ＨＧＶ５端非翻译区（５ＵＴＲ）设计引物,采用逆转录—聚合酶链反应（ＲＴ—ＰＣＲ）检测ＨＧＶＲＮＡ。结果：在８０例ｉｖ海洛因依赖者中,ＨＧＶＲＮＡ阳性率为４６．３％。结论：证实ｉｖ海洛因依赖者中ＨＧＶ感染较为常见,且吸毒时间越长,ＨＧＶ阳性率越高。     

肝病患者庚型肝炎病毒感染的临床分析

目的探讨肝病患者庚型肝炎病毒感染情况及致病性。方法应用RT-PCR方法检测312例肝病患者HGV-RNA及甲～戊型肝炎血清学标志。以HGV-RNA阳性为HGV感染判断标准,分析各型肝病患者HGV感染情况。结果312例患者HGV-RNA阳性64例,阳性率为20.51%;单纯HGV感染12例,阳性率为3.85%。结论急性肝炎,乙、丙型慢性肝炎,肝硬化,重型肝炎患者均可检出HGV-RNA。HGV多与乙、丙型肝炎病毒混合或重叠感染,亦可单纯感染引起肝脏炎症,说明HGV可能是肝病患者的致病因素之一。     

肝病患者1913例中庚型肝炎病毒感染状况的临床分析

目的　了解庚型肝炎病毒 (HGV)在各种类型病毒性肝炎中的感染状况 ,对 HGV感染的病例进行临床分析。方法　对 1913例不同临床类型的住院病人进行回顾性分析。结果　共检出抗 -HGV阳性 15 7例(8.2 1% ) ,其中单独抗 -HGV阳性者 3 6例 (1.88% ) ,与其他各型肝炎病毒均有重叠 /混合感染 ,共检出 12 1例(6.3 3 % )。在感染病例中以中青年为主 (5 9.2 4% ) ,肝癌患者中 HGV的感染率 (3 3 .3 3 % )明显高于其他组 (P<0 .0 0 5 )。结论　 HGV病毒可与其他各型肝炎病毒重叠 /混合感染 ,也可单独存在 ,可致各种肝炎、肝硬化 ,甚至可能引起肝癌     

丙型、丁型、庚型肝炎与原发性肝癌的相关因素分析

目的 探讨由RNA病毒引起的丙型、丁型、庚型肝炎与原发性肝癌(PLC)发生的关系。方法 采用免疫病理学观察,比较丙型肝炎病毒NS_5抗原(HCV-NS_5),丁型肝炎病毒抗原(HDVAg),庚型肝炎病毒NS_5抗原(HGV-NS_5)在肝细胞肝癌(HCC)组织中的表达特点。结果 在72例HCC中HCV-NS_5,HDVAg,HGV-NS_5的检出率分别为26.39％、19.44％、6.94％。总检出率为29.17％。3种病毒抗原信号在癌组织及癌旁组织中均有表达,虽其表达的主要部位有差异。3种病毒抗原的表达在HCC的分化程度方面无明显相关性,而与HCC患者的预后有一定的相关性。结论 HCV和HDV与PLC的发病关系密切,没有证据表明HGV是PLC发生的高危因素。     

庚型病毒性肝炎临床和病理初步探讨

采用反转录聚合酶链反应，从40例输血后丙型肝炎和30树非甲非乙非丙非丁非戊型肝炎（HNA－E）患者的血清中检测庚型肝炎病毒核酸（HGVRNA），阳性率分别为10．0％（4／40）和6．7％（2／30）。对6例庚型肝炎的临床和病理初步观察提示该病具有以下特点：（1）主要通过输血感染；（2）很少出现黄疸，ALT多在100U／L以下；（3）临床症状较轻，多系体检发现，（4）一般降酶护肝药很难奏效，病情容易反复；（5）慢性化发生率较高；（6）肝脏病理改变较轻，多属CPH。     

庚型肝炎临床及流行病学分析

对３３例经聚合酶链反应（ＰＴ－ＰＣＲ）法检测ＨＧＶ－ＲＮＡ阳性的庚型肝炎患者进行临床和流行病学分析。结果发现经血传播１０例（３０．３％），散发性病例２３例（６９．７％）。单纯ＨＧＶ感染６例（１８．２％），ＨＧＶ与ＨＡＶ、ＨＢＶ、ＨＣＶ二重、三重感染者２７例（８１．８％）。临床诊断急性肝炎１６例，慢性肝炎９例，重症肝炎１例，肝硬化５例，肝癌２例。单纯ＨＧＶ感染症状较轻，有轻重黄疸、ＡＬＴ轻度增高，可导致慢性肝炎，肝硬化。结果提示输血是ＨＧＶ感染的传播途径之一，还有其他传播途径，有待进一步研究。ＨＧＶ与ＨＢＶ重叠感染时可能引起重症肝炎、肝癌，应引起注意     

Occult hepatitis B virus infection in dialysis patients: a multicentre survey

BACKGROUND: The epidemiology and clinical significance of occult hepatitis B virus infection (serum hepatitis B surface antigen-negative patients with detectable hepatitis B virus viraemia in serum) remains controversial with only limited information about its prevalence in patients on long-term dialysis. AIM: To address the epidemiology of occult HBV infection in a large cohort of dialysis patients. METHODS: We screened a large cohort (n = 585) of Italian chronic dialysis patients; from this population, a group of hepatitis B virus surface antigen seronegative patients (n = 213) was tested by Amplicor hepatitis B virus Monitor Test to detect hepatitis B virus viraemia (hepatitis B virus-DNA) in serum. RESULTS: Occult hepatitis B virus infection was absent (zero of 213 = 0%). Persistent hepatitis B virus surface antigen carriage was less frequent than anti-hepatitis B virus core antibody (anti-hepatitis B core antigen) seropositive status in this study group [1.88% (11 of 585) vs. 36% (216 of 585), P = 0.0001]. No dialysis patients seropositive for anti-hepatitis B core antibody in serum (zero of 123 = 0%) had detectable hepatitis B virus-DNA by polymerase chain reaction technology. No significant association between abnormal biochemical liver tests and serum anti-hepatitis B core antibody was noted in our population. Nominal logistic regression analysis demonstrated an independent and significant relationship between anti-HCV antibody and anti-hepatitis B virus core antibody in serum (Wald chi-square 16.06, P = 0.0001). The rate of seropositive patients for anti-hepatitis B virus core antibody was higher among study patients than controls with normal renal function [36.9% (216 of 585) vs. 21.4% (59 of 275), P = 0.0001]; this difference partially persisted after correction for demographic parameters, and viral markers. CONCLUSION: In conclusion, occult hepatitis B virus was absent in our study group. Anti-hepatitis B core antibody was significantly related to presence of anti-HCV antibody supporting shared modes of transmission. Clinical studies based on molecular biology techniques provided with higher sensitivity are planned.     

The prevalence of antibodies to hepatitis C virus in patients with chronic liver disease.

OBJECT: to determine the prevalence of antibodies to hepatitis C virus in selected groups of patients with chronic liver disease. METHODS: serum specimens were obtained from 39 patients with chronic liver function abnormalities of uncertain cause (group A), from 15 patients with autoimmune chronic active hepatitis (group B) and from 10 patients with chronic hepatitis B (group C). In an extension of the study, serum was collected from sexual partners of patients found to be HCV seropositive. A second generation ELISA assay (Abbott) was used to analyse the specimens. RESULTS: ten patients (26%) in group A were seropositive, one (7%) in group B and three (30%) in group C. Risk factors for infection included blood transfusion in three, intravenous drug use in six (including the only positive patient in group B) and both factors in another patient. Only one of the 10 sexual partners tested was positive but this subject was also an intravenous drug user. CONCLUSIONS: hepatitis C virus is a significant cause of chronic liver disease in Christchurch. Important risk factors include blood transfusion and intravenous drug use although sporadic cases occur. Transmission to sexual partners is uncommon. The second generation assay does not appear to give false positive results in autoimmune chronic active hepatitis.     

GB virus C/hepatitis G virus infection in chronic hepatitis C patients with and without interferon-alpha therapy.

GB virus C/hepatitis G virus (GBV-C/HGV) RNA, detected by polymerase chain reaction, and antibodies to the GBV-C/HGV envelope protein (anti-E2), detected by an enzyme-linked immunosorbent assay, were used to evaluate both the impact of GBV-C/HGV on the coexistent hepatitis C virus (HCV) infection and the course of GBV-C/HGV infection in chronic hepatitis C patients with and without interferon-alpha (IFN-alpha) treatment. Of the 162 chronic hepatitis C patients treated with INF-alpha, 17.9% were GBV-C/HGV RNA-positive and 18.5% anti-E2-positive (total exposure, 35.2%). Neither present nor past GBV-C/HGV infection had impact on the clinical features, HCV virological characteristics and response to IFN-alpha treatment in chronic hepatitis C patients. Among patients with ongoing HCV/GBV-C/HGV coinfection, 20.7% (6/29) in IFN-alpha-treated patients lost GBV-C/HGV RNA concomitant with anti-E2 seropositivity, which was significantly higher than 4.8% (2/42) in patients without INF-alpha treatment (P<0.05). Based on multivariate analyses, the significant factors associated with clearance of GBV-C/HGV viremia combined with anti-E2 seropositivity were baseline anti-E2 seropositivity and IFN-alpha treatment. In summary, GBV-C/HGV did not alter the course of coexistent HCV. IFN-alpha treatment was effective in some patients against GBV-C/HGV and might facilitate anti-E2 seroconversion in chronic hepatitis C patients with GBV-C/HGV viremia.     

6例河南株HGV NS5基因变异的研究

目的 探讨河南省庚型肝炎病毒基因的变异特征。方法 利用逆转录－巢式－聚合酶链反应（ＲＴ－Ｎｅｓｔ－ＰＣＲ）扩增ＨＧＶ ＮＳ５区序列;将ＰＣＲ扩增产物克隆入Ｔ载体,进行测序和分析。结果 河南株ＨＧＶ ＮＳ５核酸和氨基酸序列与ＧｅｎＢａｎｋ中ＨＧＶ对应位置序列的同源笥分别为８９％￣９８．４％和９５％￣９８．４％;河南株有独特变异点（２３位Ａｌｇ→Ｇｌｙ,１１９位Ｐｈｅ→Ｌｅｕ,１５３位Ａｓｐ→Ｇｌｕ）