HGV/GBV-C感染在肝细胞损伤中的作用探讨

目的 研究单一HGV/GBV-C感染者肝组织中该病毒核酸定位及相关抗原表达,探讨HGV/GBV-C感染在肝细胞损伤中的作用。方法 12例单一HGV/GBV-C感染者经肝穿获取的肝组织常规病理诊断,采用地高辛标记探针原位杂交法检测病毒RNA,并用免疫组织化学法检测病毒相关抗原表达。结果 病理诊断急性肝炎8例,慢性肝炎4例。HGV/GBV-C NS5抗原检出阳性率为66.67％(8/12),阳性信号主要位于肝细胞胞浆中;HGV/GBV-C RNA检出阳性率为58.33％(7/12),阳性信号位于胞浆,分布无一定规律,阳性细胞与肝细胞变性、淤胆、炎性细胞浸润、细胞坏死程度等并无相关关系。单一HGV/GBV-C感染者临床表现轻,不易被发现。结论 HGV/GBV-C RNA并不直接损害肝细胞;肝细胞中存在HGV/GBV-C相关抗原表达,其编码产物可能作为一种靶抗原,诱发免疫病理反应。     

急性病毒性肝炎患者中庚型肝炎病毒RNA检测研究

为观察庚型肝炎病毒在急性病毒性肝炎中感染情况,对239例急性肝炎患者的血清,采用两步法筛选进行HGV RNA的检测,并经HGV RNA确证试验,结果显示:HGV RNA阳性者9例,其中合并乙肝1例,合并丙肝3例,合并戊肝1例,非甲-戊型肝炎中占4例。提示:①HGV与其它型肝炎病毒重叠感染较多。②6例患者无输血史,而HGV RNA仍为阳性,提示HGV的输血外传播。③NA-E急性肝炎患者中,HGV的阳性率为23.5％,证实HGV为其病原之一,同时提示还可能存在其它的致病因子。     

庚型肝炎病毒感染对急性肝炎致病性的探讨

目的 探讨急性肝炎患者肝组织中庚型肝炎病毒（ＨＧＶ）的感染状况及其致病性。方法 采用免疫组织化学方法对３７例血清学肝炎病毒标记甲￣戊型均阴性的急性肝炎患者肝穿肝组织中的ＨＧＶ ＮＳ５抗原等进行检测,并对ＨＧＶ感染的致病性进行研究。结果 ＨＧＶ ＮＳ５抗原的检出率为３７．８％（１４／３７）,其中ＨＧＶ ＮＳ５单项阳性４例（急性庚型肝炎组）。结果 ＨＧＶ ＮＳ５抗原的检出率为３７．８％（１４／３７）,     

急性重型肝炎患者肝组织中庚型肝炎病毒的检测

检测急性重量型肝炎患者肝组织中庚型肝炎病毒（ＨＧＶ）的存在状况及探讨与发病的关系,采用免疫组化方法以抗－ＨＣＶＮＳ５单克隆抗体对２６例急性重型肝炎患者尸检肝组织中的ＨＧＶ等抗原进行了检测,结果显示２６例肝组织中检测出ＨＧＶ阳性６例（２３．１％）ＨＧＶＮＳ５Ａｇ阳性着色颗粒表达于残存的肝细胞浆内,阳性细胞呈片簇状分布于汇管区周围,６例中４例重叠有ＨＢＶ或／和ＨＣＶ感染,肝组织中ＨＢｓＡｇ或／和ＨＣＶ     

单纯HGV感染急性肝炎患者的临床与病理研究

目的 通过对一组急性ＨＧＶ感染患者的肝组织病理和临床资料分析，探讨庚型肝炎病毒感染的致病性，方法 采用免疫组织化学技术对５９例不明原因包性肝炎患者的肝组织，进行乙、丙和庚型肝炎病毒怕检测，部分病例经原位杂交证实。结果 ＨＧＶＮＳ５怕的检出率为（５７．６％〈３４／５９），其中２０直组织中ＨＢｓＡｇ和／或ＨＣＶＮＳ３Ａｇ同时阳性ＩＨＧＶ合并感染组），ＨＧＶ阳性信号主要位于肝细胞浆内。ＨＧＶ单纯感染病例     

A high frequency of GBV-C/HGV coinfection in hepatitis C patients in Germany

AIM:To detect infection rate of GBV-C/HGV in hepatitis C patients, to determine the methods of higher sensitivity and the primers of higher efficiency for GBV-C/HGV RNA detection and to study the dominant subtype and mutation of GBV-C/HGV.METHODS:Quantitative RT-PCR for detection pf HCV RNA concentration in serum samples, RT-nested PCR with two sets of primers for detection of GBV-C RNA, RT-PCR ELISA with two sets of primers for detection of HGV RNA, nucleotide sequence and putative amino acid sequence analysis.RESULTS:The positive rates of GBV-C RNA at the 5'-NCR and NS3 region in 211 serums amples from the patients with HCV infection were 31.8% and 22.8% respectively. The positive rates of HGV RNA at the 5'-NCR and NS5 region in the same samples were 47.9% and 31.8% respectively. The total positive rate of GBV-C/HGV RNA was as high as 55.5%. HCV copy numbers in the patients without GBV-C/HGV coinfection were statistically higher than that in the patients with GBV-C/HGV coinfection (P<0.01).Frequent mutation of nucleotide residue was present in the amplification products. Frameshift mutation was found in two samples with GBV-C NS3 region nucleotide sequences. All nucleotide sequences from amplification products showed higher homology to HGV genome than to GBV-C genome even though part of the sequences were amplified with GBV-C primers.CONCLUSION:A high frequency of GBV-C/HGV coinfection existed in the hepatitis C patients. RT-PCR ELISA was more sensitive than RT-nested PCR for detection of GBV-C/HGV RNA. The primers derived from the 5'-NCR was more efficient than those derived from the NS3 and NS5 regions. A reverse relationship was found to exist between HCV RNA concentration and GBV-C/HGV infection frequency. HGV was the dominant subtype of the virus in the local area. The major mutations of GBV-C/HGV genomes were random mutation of nucleotide residue.     

慢性肝病患者肝组织中HGV抗原的表达及意义

目的探讨慢性肝病患者组织中庚型肝炎病毒（ＨＧＶ）表达与意义。方法应用免疫组织化学方法以鼠抗ＨＧＶＮＳ５甲克隆抗体检测１４２例慢性肝病患者肝组织中ＨＧＶ抗原，部分患者采用ＲＴ－ＰＣＲ方法检测其血清中ＨＣＶＲＮＡ。结果１４２例肝病患者中，２９例（２０．４％）组织中检出ＨＧＶ抗原，肝硬化组（４２．９％，１２／２８）较慢性肝炎（１５．９％８／１１）和肝癌（１４．３％，９／６３）组检出率高。阳性信号位于胞浆中，阳性细胞可成散在、簇状或弥漫性分布。阳性细胞周围可有炎性坏死；肝癌患者抗原阳性细胞主要位于癌旁肝组织，癌巢中仅偶见少数散在分布的阳必细胞；绝大多数组织抗原阳性者其血清ＨＧＶＲＮＡ为阳性。有４例患者组织中ＨＧＶ抗原阳性但其血清ＨＧＶＲＮＡ阴性。结论ＨＧＶ可在慢性肝病包括肝细胞肝癌患者肝组织中表达，ＨＧＶ感染在慢性肝炎病期进展及肝癌发生中具有一定意义。     

Prevalence of hepatitis G virus in liver disease.

The prevalence of hepatitis G virus (HGV) in liver disease of non-A, -B, -C viral hepatitis, hepatitis B and hepatitis C was determined. Two of 44 patients (4.5%) with liver injury without any hepatitis A, B or C marker were positive for HGV. One of five cases of hepatocellular carcinoma was positive for HGV. One of three cases with fulminant hepatitis was positive for HGV. This case was negative at the onset of fulminant hepatitis and became positive after plasmapheresis. No patient with acute (n=8) or chronic (n=5) hepatitis or liver cirrhosis (n=8) was positive for HGV in non-A, -B, -C liver disease. One of 30 patients with various HBV-positive liver diseases and nine (17.3) of 52 patients with type C liver disease were positive for HGV. In patients with hepatitis C, four (28.6%) of 14 HGV-co-infected patients were complicated with diabetes mellitus compared with four (10.5%) of 38 single hepatitis C virus (HCV)-infected patients (not significant). In 12 HGV-positive patients, eight of 10 (80%) had a history of blood transfusion. In HCV-positive patients, co-infection with HGV was not a risk factor in patients with diabetes mellitus as a complication. HGV appeared to cause non-A, -B, -C hepatitis rarely, and its main route of infection was blood transfusion.     

Hepatitis G Virus Infection Among Liver Graft Recipients: Anatomoclinical Correlations

Hepatitis G virus (HGV) causes persistent infection in man, but its disease association is controversial. We studied the HGV disease association in 25 liver transplantation (LT) recipients without evidence of hepatitis B and C infection. HGV RNA was tested by semiquantitative RT-PCR in serial serum samples and its presence was correlated with the biochemical and histological evidence of liver damage. The overall prevalence of HGV infection in this population was 9/25 (36%), one patient being HGV RNA positive since before LT, while the other eight apparently acquired de novo infections after LT. In five cases, appearance of HGV was followed by biochemical and histological evidence of liver damage: the liver biopsy showed acute rejection in two cases, acute cholangitis in two, and acute hepatitis in one. At the end of follow-up, histological evidence of chronic hepatitis was found in one HGV-positive patient but also in three HGV-negative patients, whereas the only patient with acute hepatitis at the time HGV RNA was first detected in serum developed an intralobular gigantocellular granuloma. In conclusion, HGV infection after LT may be seldom associated with acute and chronic liver damage, but comparable histological features can be observed also among HGV-negative controls.     

庚型肝炎患者肝组织中HGV表达的免疫组化研究

目的探讨庚型肝炎病毒（ＨＧＶ）在庚型肝炎肝组织中的表达状况与临床意义．方法应用免疫组织化学ＰＡＰ方法以鼠抗ＨＧＶＮＳ５单克隆抗体对庚型肝炎患者２０例（急性肝炎２例，慢性肝炎８例，肝硬变１０例，血清ＨＧＶＲＮＡ皆阳性）肝组织中ＨＧＶ抗原进行检测．结果庚型肝炎患者２０例中，８例（４０％）肝组织中检出ＨＧＶ抗原；不同病期检出率分别为：急性肝炎０／２（０％），慢性肝炎２／８（２５％），肝硬变６／１０（６０％），各组间差异无显著意义；阳性信号位于肝细胞胞质；阳性细胞可位于炎症坏死灶周围；抗原阳性与阴性组间肝组织炎症活动度及血清谷丙转氨酶水平无明显差别，但阳性组纤维化指数较高．结论ＨＧＶ感染及其在肝组织中表达可能与肝组织纤维化有一定关系     

地高辛素探针原位杂交检测非甲—非戊型肝炎肝组织中HGV RNA

目的观测ＨＧＶＲＮＡ在非甲－非戊型急慢性病毒性肝炎肝组织中的表达，对ＨＧＶ感染与急慢性病毒性肝炎发病的关系进行探讨。方法以地高辛素标记ＨＧＶｃＤＮＡ探针，对３６例血清非甲－非戊型急性和慢性病毒性肝炎患者石蜡包埋肝穿肝组织进行原位杂交，并对急性ＨＧＶ感染病例进行了随访观察。结果３６例肝病患者中，１１例（３０６％）肝组织中检出ＨＧＶＲＮＡ。其中急性重症肝炎１例（１／３），亚急性重症肝炎３例（３／６），急性轻型肝炎５例（５／１６）慢性肝炎２例（２／１１）。阳性信号表达于肝细胞胞浆及／或核内，在急性轻型肝炎，阳性信号多弥漫散在分布。而在慢性肝炎，则多呈片灶状靠近汇管区周围。在检出率和阳性细胞分布方面，原位杂交与免疫组化检测结果（３６１％，１３／３６）基本一致。８－３８个月后的急性轻型肝炎随访二次肝穿检测显示了肝组织中ＨＧＶＲＮＡ持持续存在。结论在急慢性非甲－非戊型病毒性肝炎肝组织中，ＨＧＶ感染较为常见，ＨＧＶ感染可能与急性病毒性肝炎的发病及慢性化有关。     

HGV/GBV-C infection in patients with acute hepatitis of different etiology and in patients with chronic hepatitis C

To investigate the prevalence of hepatitis G virus (HGV/GBV-C) in patients with liver disease and to confirm its hypothesized ability to cause liver damage, we studied 130 subjects; 61 had chronic hepatitis C virus infection and 69 had acute hepatitis of either defined etiology (n = 57) or of unknown origin (n = 12). Positivity for HGV/GBV-C RNA was detected in 10 of the 61 subjects with chronic hepatitis C (16.3%) and in 11 of the 57 subjects with acute hepatitis of defined etiology (19%), whereas we failed to detect HGV/GBV-C viremia in subjects with hepatitis of non-established etiology. Patients exhibiting positivity for HGV/GBV-C RNA were found to be comparable to those exhibiting negativity for HGV/GBV-C RNA in terms of both liver function tests and Knodell's score (in liver biopsies); the affect of HGV/GBV-C infection on the biohumoral and histological activity in patients with chronic hepatitis C therefore appears to be minimal or absent. Similar clinical features were observed in patients with acute hepatitis of known etiology whether they were positive or negative for HGV/GBV-C RNA. However, long-term clinical studies are still required to clarify the actual impact of HGV/GBV-C co-infection. In our geographic, i.e., a region or north-east Italy, HGV/GBV-C infection appears to be strictly related to intravenous drug use, and this agent does not seem to be responsible for acute hepatitis of unknown etiology; other etiological agents are probably involved. Received Feb. 17, 1997; accepted June 27, 1997     

庚型肝炎病毒致病性的临床研究

目的:调查庚型肝炎病毒(HGV)在各型病毒性肝炎中的感染率和探讨其对临床可能的影响。方法:以巢式逆转录PCR法检测1997年1月-1999年12月住院的5024例患者血清中HGV 5’NCR和NS5b基因片段,两者均检出判定为HGV感染,同时与临床类型、肝功能、疾病转归进行比较。结果:①5024例肝病患者中550例检出HGV RNA,总的感染率为10.9％,其中慢性肝炎、重型肝炎的感染明显高于急性非甲-戊型、急性甲型和急性乙型肝炎(14.2％和15.8％ vs 7.8％、4.6％、8.3％,P<0.05)。②急性戊型肝炎合并HGV感染率高达14.2％,明显高于其他急性病毒性肝炎(P<0.05)。③重叠HGV感染与否并不影响各型肝炎的肝功能损害程度,也不影响重型肝炎患者的存活率(61.43％ vs 59.85％,P>0.05)。④27例单一感染HGV的惠性肝炎患者。均临床治愈出院,HGV的病毒血症与肝炎的临床表现没有相关性,37％(10/27)患者在6个月随访期中在肝功能恢复正常后仍有HGV病毒血症。结论:①HGV感染可发生于各型病毒性肝炎患者。②急性戊型肝炎合并HGV感染明显高于其他急性病毒性肝炎的原因,值得进一步探讨。③HGV感染对人类没有重要的致病性,HGV感染不是原因不明的非甲-戊型肝炎的主要病因。     

A study on pathogenicity of hepatitis G virus

AIM: To study the pathogenicity of hepatitis G virus (HGV) and observe the genesis and pathological process of hepatitis G. METHODS: HGV-RNA in serum was detected by RT-PCR assay. The immunohistochemical assays of liver tissue were performed with HGV monocoloned antibody (McAb) expressed from the region of HGV NS5 nucleic acid sequence. The clinical and pathological data of 52 patients with hepatitis G were discussed. In animal experiment, the Chinese Rhesus monkeys were infected with the serum of a patient with HGV infection. And the dynamic changes in serology and liver histology of animals were observed. RESULTS: One hundred and fifty-four patients with HGV-RNA positive were selected from 1552 patients with various kinds of hepatitis. Of 154 patients with HGV infection, 52 were infected with HGV only, which accounted for 33.8 (52/154) and 102 with positive HGV-RNA were super-infected with other hepatitis viruses, which accounted for 66.2 (102/154). The clinical and pathological observation showed that the acute and chronic hepatitis could be induced by HGV. The slight abnormality of transaminases ALT and AST in serum of monkeys lasted nearly 12 months and histological results showed a series of pathological changes. CONCLUSION: HGV is a hepatotropic virus and has pathogenicty.