Kozo Fujio

A prospective and comparative cohort study on efficacy and drug resistance during long-term lamivudine treatment for various stages of chronic hepatitis B and cirrhosis

Background and Aims:  A prospective, non‐randomized cohort study on long‐term lamivudine treatment, comparing efficacy, drug resistance, and prognosis for various stages of chronic hepatitis B virus (HBV)–related liver disease was performed to elucidate the significance and indication of lamivudine for individual patients at each stage of disease.

Expression of Perforin and Fas Ligand mRNA in the Liver of Viral Hepatitis

Cytotoxic T lymphocytes (CTLs) play an important role in the pathogenesis of viral hepatitis. We studied the expression of mRNAs of perforin and Fas ligand (Fas-L) in biopsy specimens from chronic hepatitis B (CHB) (15 cases) and hepatitis C (CHC) patients (13 cases). Both perforin and Fas-L mRNAs were detected in all cases of both CHB and CHC. No messages were detected in the control livers from two cases of fatty liver, a case of Gilberts syndrome, and a case of Dubin–Johnson syndrome. Semiquantitative analysis revealed a positive correlation between the intensity of perforin and Fas-L mRNAs in both CHB and CHC. In CHB, the intensity of both perforin and Fas-L mRNAs showed a positive correlation with the histological activity and serum alanine aminotransferase level, while the correlation was not apparent in CHC. These results suggest that both perforin and Fas/Fas-L systems are involved in the pathogenesis of liver cell injury of CHB and CHC.

Hepatitis B Virus Gene in Liver Tissue Promotes Hepatocellular Carcinoma Development in Chronic Hepatitis C Patients

The hepatitis B virus (HBV) gene has been detected in hepatocellular carcinoma (HCC) tissue negative for the hepatitis B surface antigen and positive for the hepatitis C virus (HCV) antibody, but the precise role of the HBV gene in hepatocarcinogenesis has yet to be clarified. We studied the HBV gene in liver tissue several years before the emergence of HCC. Eleven patients diagnosed with HCV-positive chronic liver disease and who developed HCC were assigned to group A. HBV DNA was detected in 8 of the 11 patients (73%). Twenty-five patients, who did not develop HCC, were selected as group B. Six of the group B patients were classified as DNA-positive (24%). The HBV DNA in liver tissue was found to be significantly related to HCC development (P < 0.01). Thus, the presence of the HBV gene in patients with chronic HCV associated-liver injury appears to promote hepatocarcinogenesis, although prospective studies are needed to confirm this result.

Possible contribution of prior hepatitis B virus infection to the development of hepatocellular carcinoma

Background:  The prevalence of prior hepatitis B virus (HBV) infection in hepatocellular carcinoma (HCC) patients and its role in hepatocarcinogenesis are not clear. The aim of the present study is to clarify the importance of prior HBV infection in development of HCC.

Two cases of chronic hepatitis B with emergence of lamivudine-resistant virus during long-term therapy

Abstract Lamivudine is a potent antiviral drug for chronic hepatitis B, however the emergence of mutant virus resistant to lamivudine has been reported in patients with long-term administration of this drug. Here we report two patients with chronic hepatitis B treated with 48- and 52-weeks administration of lamivudine whose serum hepatitis B virus (HBV) DNA became positive during therapy. The amino acid sequence of the polymerase region of HBV from these patients revealed a YI/VDD mutation of the YMDD motif and a 223Leu to Met mutation, which were reported previously in immune suppressed patients during long-term lamivudine therapy for hepatitis B. After the cessation of therapy, serum HBV-DNA returned to wild-type immediately. Emergence of mutant virus resistant to lamivudine should be monitored during long-term lamivudine therapy.

Detection of serum RNA specific for blood-borne non-A, non-B hepatitis.

After years of efforts to identify etiologic agent(s) for bloodborne non-A, non-B hepatitis (BNANBH), several groups of investigators, including ours, have isolated clones of cDNA specific for BNANBH(1). Here we present a preliminary report on the method to detect serum RNA specific for BNANBH by the slot-blot hybridization. [Materials a~d M~thQds] RNA was extracted by AGTC method(2) from the precipitates from 4ml-aliquots of serum with 4% PEG-4000(w/v). RNA was denatured with 7.4% formaldehyde and slot-blotted on nitrocellulose filter, followed by baking at 80~ for 2 hours. Antisense RNA probe was transcribed in vitro by T7 RNA polymerase from pHSNI4-3 shown in Fig.l. 32p-CTP was used for radiolabelling. Hybridization was performed in solution including 50% formamide, 5X SSPE, 5X Denhardt, 1% SDS, at 60~ After washing, nonspecifically bound RNA probe was digested by l~g/ml RNase A at 37~ for 15 rain. Filters were autoradiographed at -70~ with intensifying screens. [Results] 0.1pg of sense RNA can be detected by this method. Total RNA from neither E. coli nor normal human liver hybridize with this probe by Northern-blot hybridization. While serum RNA extracted from patients with post-transfusion non-A, non-B hepatitis were positive for NI4, RNA from patients with acute hepatitis A or B were negative (Fig. 2) . This method will be clinically useful for the diagnosis of BNANBH.