Evaluation of serum concentrations of human hepatocyte growth factor during interferon therapy for chronic hepatitis C.

In this study, the serum concentrations of human hepatocyte growth factor (HGF) were examined to clarify the relationship between HGF and interferon (IFN) therapy for hepatitis C. The subjects were 94 patients with chronic hepatitis C who underwent liver biopsy at our institution from 1994 through 1996. These patients were treated with natural IFN-alpha, IFNalpha(2a) and IFNalpha(2b) for periods varying from 12 to 26 weeks. Serum levels of HGF were determined in individual patients just before and after the administration of IFN and at 6 months and 1 year thereafter. The serum concentration of HGF was evaluated using enzyme-linked immunosorbent assay kits. The intra-hepatic location of HGF was explored using an immunoperoxidase-staining method. A positive correlation was found between the degree of HGF expression in the liver and the serum HGF concentrations. The degree of HGF expression in the liver decreased in the virologically sustained responders (SVRs) following IFN therapy. The serum HGF concentrations immediately after IFN therapy were lower than those before therapy in 83% of the patients. The concentrations gradually rose thereafter in about 45% of the non-responders, while it remained low or declined further in about half of the patients in this group. In the SVRs, the serum HGF concentrations declined in 88% of patients immediately after IFN therapy. Thereafter, it remained equally low or declined further in 60% of the SVRs. The serum HGF concentrations at 6 months and 1 year after IFN therapy were significantly lower in the SVRs than in the non-responders. In conclusions, serum HGF concentrations declined following IFN treatment regardless of the virological outcome of treatment. The decrease in serum HGF concentrations results from a decrease in the number of mesenchymal cells producing HGF. Consequently, evaluation of the serum HGF concentration is of clinical value for assessing changes in liver tissues after IFN therapy.

Full genomic analysis of hepatitis delta virus prevalent on Miyako Island, Japan.

The aims of this study were to determine the full-length genome sequences of hepatitis delta virus (HDV) in HDV RNA-positive subjects, and to elucidate the molecular specificity of the HDVs that are clustered on a distant island in Japan. This study included 3 subjects with chronic hepatitis who were positive for hepatitis B surface (HBs) antigen and HDV RNA, and who were admitted to the Okinawa Prefectural Miyako Hospital in 1998. The full-length genome sequence of HDV was determined by nested polymerase chain reaction (PCR) using four kinds of primer sets. The genomic length of HDV was 1,675, 1,679 and 1,681 base pairs, respectively. There was 90-92% nucleotide homology between each pair of isolates. In comparison with HDV isolates in geographically neighboring regions, the nucleotide homology of the 3 HDV isolates were 73-75% with the China isolate of genotype I; 77-78% with the Taiwan isolate of genotype I; 83-84% with a Japan isolate of genotype IIa; 85-87% with the Taiwan isolate of genotype IIa, and 87-88% with the Taiwan isolates of genotype IIb. Therefore, the Miyako Island isolates had high homology with the Taiwan isolate of genotype IIa and IIb. Phylogenetic analysis of the full-length genome sequences of HDV revealed that the two Miyako Island isolates were classified into a genotype IIb'. The other one was classified as genotype IIb. In conclusion, the HDV of Miyako Island isolates can be classified as a novel subgroup of genotype IIb, designated type IIb', and genotype IIb.

SEN virus infection influences the pathological findings in liver but does not affect the incidence of hepatocellular carcinoma in patients with chronic hepatitis C and liver cirrhosis.

BACKGROUND/AIMS: This investigation compared the histological findings in the livers of chronic hepatitis C patients who were or were not co-infected with SEN virus (SEN-V) to determine the histological and clinical characteristics of SEN-V infection in Japan. METHODS: Three hundred and ninety-two patients with hepatitis C virus-associated chronic hepatitis (CH) or liver cirrhosis (LC) were included in the study. Serum samples were tested for the presence of SEN-V DNA by nested polymerase chain reaction. The liver biopsy specimen of each patient was examined and scores were assigned to indicate the severity of each of the following features: inflammatory cell infiltration in the periportal, parenchymal, and portal areas; F stage; portal sclerotic change; perivenular fibrosis; pericellular fibrosis; damage to the bile ducts; steatosis and irregular regeneration of hepatocytes (IR). RESULTS: Of the 473 patients, 194 (41.0%) were positive for SEN-V DNA. The rate of progression of F stage correlated with SEN-V DNA positivity. The blood biochemical parameters did not differ significantly between the SEN-V DNA-positive and -negative patients. The histological features of the livers of SEN-V DNA-positive patients included more severe parenchymal inflammatory cell infiltration and more IR. In particular, among those at the F2, F3 and F4 stages, the degree of IR of the SEN-V DNA-positive patients was significantly greater than that of the SEN-V DNA-negative patients. The cumulative probability of hepatocellular carcinoma (HCC) incidence and survival rate did not differ between the SEN-V DNA-positive and-negative patients. CONCLUSIONS: SEN-V co-infection may influence the histopathological features of the livers of patients with type C CH and LC but does not affect the outcome of patients with type C chronic liver disease.

Decreased risk of hepatocellular carcinoma in patients with chronic hepatitis C whose serum alanine aminotransferase levels became less than twice the upper limit of normal following interferon therapy.

AIM: The incidence of hepatocellular carcinoma (HCC) in C-viral chronic hepatitis (CH) and liver cirrhosis (LC) patients after interferon (IFN) therapy was evaluated according to alanine aminotransferase (ALT) levels. PATIENTS: Two hundred sixty-nine patients with C-viral CH and LC were treated with natural IFN-alpha. The efficacy of IFN therapy was evaluated based on virologic response and ALT levels using the following groups: virologic-sustained responders (VSR); biochemical-sustained responders (BSR); partial responders (PR), which consisted of BSR patients whose serum ALT levels later relapsed; non-responders (NR)1, which included patients with serum ALT levels that were usually less than 80 IU/l; and NR2, NR with ALT levels persistently more than 80 IU/l. RESULTS: Of the 269 patients, 22 (8.2%) developed HCC after IFN therapy. The incidence of HCC (%/patient/year) was 0.78%, 0%, 0%, 0.17%, 4.68% in VSR, BR, PR, NR1, NR2, respectively. Multivariate analysis revealed that an increase in ALT levels to more than 80 IU/l is an important risk factor for the occurrence of HCC. CONCLUSIONS: We concluded that the patients with ALT levels less than twice the upper limit of normal after IFN therapy have a reduced risk of progression to HCC from C-viral chronic liver disease.

Detection of serum and intrahepatic KL-6 in anti-HCV positive patients with hepatocellular carcinoma.

We investigated the clinical significance of serum and intrahepatic KL-6/MUC1 (KL-6) in patients with hepatitis C virus (HCV) antibody-positive hepatocellular carcinoma (HCC). The subjects included 76 patients diagnosed with anti-HCV positive HCC, 69 with, and 51 without, liver cirrhosis (LC). Frozen serum samples were obtained from each subject to determine the serum KL-6 levels using an enzyme-linked immunosorbent assay. Expression of KL-6 antigen in the liver was also investigated using immunoperoxidase staining. The mean serum KL-6 level in patients with HCC was [Formula: see text] U/ml (319U/ml for HCC with LC, 342.8U/ml for HCC without LC). Serum KL-6 levels in patients with HCC with LC and HCC without LC did not differ. Serum KL-6 levels were elevated with increases in the size of spaces occupied by tumors in the liver. Among patients with HCC, there was no correlation between serum KL-6 levels and alpha-fetoprotein (AFP) levels and protein induced by vitamin K absence or antagonist-II (PIVKA-II) levels. However, some patients with low levels of AFP and PIVKA-II possessed high levels of KL-6. Furthermore, serum KL-6 levels decreased after therapy for HCC nodules. Immunohistochemical staining showed KL-6 antigen was detected within the cell membrane and in the cytoplasm of cancer cells. KL-6 antigen was localized on the membrane and the endoplasmic reticulum of cancer cells in the cancerous foci by electron microscopy. Our results suggest that serum KL-6 levels represent a serological marker of HCC development, because KL-6 expression was localized to the cancer cells in HCC nodules.

Long-term outcome, with monitoring of platelet counts, in patients with chronic hepatitis C and liver cirrhosis after interferon therapy.

OBJECTIVE: Because the determination of the stage of fibrosis depends on rather subjective judgment, more objective parameters are needed. In this study, we followed the long-term outcome, with monitoring of platelet counts, in patients with chronic hepatitis C or liver cirrhosis (LC) who had undergone interferon (IFN) therapy. METHODS: 596 patients who were diagnosed at our institute from 1987 to 1998 with chronic hepatitis C and LC were treated with IFNs. A further 58 patients were not treated (NT). The annual rate of changes in platelet counts were calculated and compared for IFN-treated and NT patients. RESULTS: The relationship between the efficacy of IFN therapy and the incidence of hepatocellular carcinoma (HCC) showed that the patients who were virologic sustained responders (VSR) had a significantly lower incidence of HCC than the nonresponders (NR) and NT patients. The change in platelet counts was +4,350/microl/year in the VSR, +1,010/microl/year in the biochemical sustained responders (BSR), -4,540/microl/year in the NR and -6,180/microl/year in the NT patients, indicating a significant platelet increase in the VSR, a decrease of the same magnitude in the NR and NT patients, and no change in the BSR. The cumulative probability of developing HCC and liver failure was significantly higher in groups with decreased platelet counts than in groups with increased platelet counts among patients who had undergone IFN therapy. Multivariate analyses revealed that a decrease in platelet counts was the cardinal risk factor for development of HCC and liver failure in chronic hepatitis C or LC patients. CONCLUSION: Investigation of platelet counts was useful for determining the long-term outcome of patients who had undergone IFN therapy and for predicting the development of HCC.

TT virus infection does not affect the clinical profiles of patients with hepatitis B and C in Yanbian City, China.

China is an area of high endemicity for viral hepatitis, and the molecular epidemiological investigation of TT virus (TTV) infection is of interest. In the present study, we investigated the epidemiology, clinical significance and molecular characteristics of TTV infection in patients with chronic hepatitis B and C in Yanbian City, China. Serum samples obtained from 74 patients with hepatitis B and hepatitis C who visited Yanbian Hospital, located in northeast China, were analyzed in this study. The study group included 22 cases of chronic hepatitis B (B-CH), 17 cases of liver cirrhosis B (B-LC), 7 cases of hepatocellular carcinoma (B-HCC), 16 cases of chronic hepatitis C (C-CH), 11 cases of liver cirrhosis C (C-LC) and 1 case of hepatocellular carcinoma (C-HCC). Detection of TTV DNA was performed as described by Nishizawa et al. The second-round PCR products from 7 subjects were sequenced, followed by investigation of nucleotide homology and phylogenetic analysis. TTV DNA was present in 18.2, 5.9, 28.6, 6.3, 9.1 and 0% of the patients with B-CH, B-LC, B-HCC, C-CH, C-LC and C-HCC, respectively. The highest prevalence of TTV infection was seen in the groups aged 40-50 and over 60 years. There was no significant correlation between the presence of TTV DNA and the clinical parameters in patients with hepatitis B and C. The various isolates showed 97.9-100% with isolates reported previously from Japan and 98.4-100% with isolates reported previously from China. Nucleotide sequence analysis revealed that the Yanbian isolates could be classified in the same group as the Japan and China isolates. We concluded that chronic coinfection with TTV did not affect the serological features of chronic hepatitis B and C in China, as found in Tokyo, Japan.

Analysis of background factors and evaluation of a population at high risk of hepatocellular carcinoma.

OBJECTIVE: We investigated the background clinical factors of patients with hepatocellular carcinoma (HCC) diagnosed at our institute and, from these results, determined those factors important for evaluation of a population at high risk of HCC. METHODS: This study comprised 250 patients diagnosed with HCC from 1990 through 1995 in the Nihon University Itabashi Hospital. Background clinical factors, such as the results of blood chemistry at the time of the first angiography, were examined. RESULTS: Markers of viral hepatitis in the 250 cases were as follows: type B (B-HCC), 29 (11.6%), type C (C-HCC), 201 (80.4%); type B+C, 3 (1.2%), and non-B, non-C (NBNC-HCC), 17 (6.8%). Approximately 35% of B-HCC and NBNC-HCC cases, but only 6% of C-HCC cases, exhibited platelet counts (PLT) equal to or more than 200,000/ml. On the other hand, 56.5% of the C-HCC cases exhibited PLT less than 100,000/ml, in contrast to less than 30% of the B-HCC and NBNC-HCC cases. Irrespective of the etiology of HCC, male sex and a history of smoking were characteristic risk factors. The percentages of abnormal results in combinations of tests in the B-HCC, C-HCC and NBNC-HCC subsets were: 69, 97 and 70% in the aspartate aminotransferase (AST) + PLT group (group A); 83, 98 and 70% in the group A + alanine aminotransferase (ALT) group (group B), and 86, 98 and 100% in the group B + gamma-glutamyl transpeptidase (gamma-GTP) group (group C), respectively. When hepatitis B surface antigen (HBsAg) and hepatitis C antibody (HCV-Ab) were also taken into account, abnormal results were found in every case in all of the HCC groups. CONCLUSION: C-HCC was found predominantly in cases with liver cirrhosis, whereas B-HCC and NBNC-HCC were observed more frequently in cases without liver cirrhosis. Testing for HBsAg and HCV-Ab, in addition to AST, ALT, PLT and gamma-GTP, is considered necessary when screening for HCC.

The clinical significance of serum KL-6 levels in patients with type C liver diseases.

We determine whether the serum KL-6/MUC1 (KL-6) levels in patients with type C liver disease can be used to assay inflammatory activity and the stage of fibrosis of patients, as well as to screen high-risk groups for the development of hepatocellular carcinoma (HCC). Study subjects included 130 patients with type C chronic hepatitis (CH), 15 patients diagnosed with type C acute hepatitis (AH) and 17 healthy control subjects. Frozen serum samples were obtained from each subject to determine the KL-6 levels using an enzyme-linked immunosorbent assay (EIA) method. The mean KL-6 levels in patients were as follows: 150.1 U/ml for healthy controls, 203.7 U/ml for AH patients, 225.7 U/ml for F0 stage, 207.4 U/ml for F1 stage, 235.8 U/ml for F2 stage, 193.3 U/ml for F3 stage, and 276.2 U/ml for F4 stage in CH patients. The mean serum KL-6 level in patients with F4 stage was significantly higher than that in healthy controls. No relationship was observed between the serum KL-6 levels and liver histology. However, the degree of irregular regeneration (IR) of hepatocytes and the levels of KL-6 were significantly correlated according to the progression of F stages. The cumulative incidence of HCC in the high KL-6 level group (>/=300 U/ml) was significantly greater than that in the low level group. Our results suggest that the determination of serum KL-6 levels may be useful in screening high-risk groups for the development of HCC.

Genotype analysis, using PCR with type-specific primers, of hepatitis B virus isolates from patients coinfected with hepatitis delta virus genotype II from Miyako Island, Japan.

OBJECTIVES: The aims of this study were to determine the hepatitis B virus (HBV) genotypes in hepatitis delta virus (HDV) RNA-positive patients and to characterize the HBV nucleotide sequences that may be found on a distant island of Japan. METHODS: This study included three patients with chronic hepatitis who were positive for hepatitis B surface antigen (enzyme-linked immunosorbent assay; ELISA), HDV antibody (ELISA) and HDV RNA by polymerase chain reaction (PCR). The HBV genotype was determined by nested PCR using type-specific primers. The first-round PCR products from two patients were sequenced, followed by an investigation of nucleotide homology. RESULTS: Viruses from all three patients in this study were classified as HBV genotype B. Comparison with HBV isolates from geographically neighboring regions revealed that the two HBV isolates had 97.9-98.6% identity at the nucleotide level to a Chinese isolate, 98.3-98.6% identity to the Okinawa isolate and 98.6-98.8% identity to a Japanese isolate of genotype B. On phylogenetic analysis, the HBV isolates from the two patients were classified as HBV genotype B. The HBV isolates of cases 1 and 3 clustered in the same group as isolates from the Chinese mainland and Japanese mainland, which are geographically near Miyako Island. CONCLUSION: The HBV isolates coinfected with HDV found on Miyako Island were of genotype B. The PCR method based on genotype-specific primers was useful in determining HBV genotypes.

An adult patient with acute hepatitis type B which was protracted and complicated by polyarteritis nodosa: a case report.

Polyarteritis nodosa (PAN), an extra-hepatic complication of hepatitis type B, is usually treated with a combination of immunosuppressive and antiviral drugs. Less commonly, interferon (IFN) has been used alone. A 57-year-old man was admitted to our hospital with epididymitis and acute hepatitis with genotype A, hepatitis B virus (HBV). He became a carrier with complicating PAN. The administration of interferon alpha2b (IFNalpha2b) at a dose of 6 MU/day for 4 weeks alleviated abdominal and neuronal symptoms related to PAN, although HBV replication was not eliminated. This is the first case of PAN with HBV of genotype A and which was treated with IFN alone. This case indicates the possibility that IFN is efficacious for extra-hepatic complications of hepatitis type B and suggests that differences in genotype may be associated with variation in the likelihood of progression to persistent infection and the incidence of extra-hepatic complications in Japan.

Serum concentrations of human hepatocyte growth factor is a useful indicator for predicting the occurrence of hepatocellular carcinomas in C-viral chronic liver diseases.

BACKGROUND: Numerous reports have examined the relationship between hepatocyte growth factor (HGF) and either the facilitation or suppression of the occurrence of hepatocellular carcinoma (HCC). METHODS: In this study, we measured serum HGF concentrations of blood samples and conducted prospective studies to examine the long-term outcome of C-viral chronic hepatitis (CH) and cirrhosis in patients. The subjects examined in this study include 99 patients with C-viral CH, cirrhosis, and HCC. The serum HGF level was measured in blood samples within 48 hours of collection using enzyme-linked immunosorbent assay kits. RESULTS: The serum concentrations of HGF were significantly higher in patients with HCC than in patients with CH or cirrhosis. The detection rate of HGF and its mean serum level were significantly higher in patients with a low platelet count than in patients with a high platelet count. All of the patients with serum HGF concentrations of more than 0.6 ng/mL had HCC, irrespective of the levels of alpha-fetoprotein, vitamin K absence, or antagonist-II in the blood. Serum HGF concentrations increased concomitantly with increases in areas occupied by HCC. The cumulative incidence of occurrence of HCC was significantly higher in patients with high HGF concentrations than in patients with low HGF concentrations. Multivariate analysis revealed that the elevation in serum HGF level is the most important risk factor for the occurrence of HCC. CONCLUSIONS: The serum level of HGF represents the degree of the carcinogenic state in the liver of patients with C-viral CH and cirrhosis. Therefore, the determination of serum HGF concentrations may be useful as a third tumor marker of HCC in detection as well as follow-up therapy.