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Significant associations of HLA-DP alleles with chronic hepatitis B (CHB) infection are evident in Asian and Arabian populations, including Japanese, Han Chinese, Korean, and Saudi Arabian populations. Here, significant associations between CHB infection and five DPB1 alleles (two susceptibility alleles, DPB1(*) 05:01 and (*) 09:01, and three protective alleles, DPB1(*) 02:01, (*) 04:01, and (*) 04:02) were confirmed in a population comprising of 2582 Japanese individuals. Furthermore, odds ratios for CHB were higher for those with both DPB1 susceptibility alleles than for those with only one susceptibility allele; therefore, effects of susceptibility alleles were additive for risk of CHB infection. Similarly, protective alleles showed an additive effect on protection from CHB infection. Moreover, heterozygotes of any protective allele showed stronger association with CHB than did homozygotes, suggesting that heterozygotes may bind a greater variety of hepatitis B-derived peptides, and thus present these peptides more efficiently to T-cell receptors than homozygotes. Notably, compound heterozygote of the protective allele (any one of DPB1*02:01, *04:01, and *04:02) and the susceptible allele DPB1*05:01 was significantly associated with protection against CHB infection, which indicates that one protective HLA-DPB1 molecule can provide dominant protection. Identification of the HLA-DPB1 genotypes associated with susceptibility to and protection from CHB infection is essential for future analysis of the mechanisms responsible for immune recognition of hepatitis B virus antigens by HLA-DPB1 molecules.

Many of the current reverse transcription (RT)-PCR assays for the detection of hepatitis C virus (HCV) RNA are multistep processes which use multiple enzymes and buffers. The assays are also often suboptimal, requiring nested amplification to achieve the desired levels of sensitivity. As a result, these assays are cumbersome and prone to false-positive results. The susceptibility to contamination is further aggravated by the lack of carryover controls. We have previously reported the development of a combined RT-PCR assay for HCV RNA detection which is sensitive and simple to perform. We have since successfully integrated dUTP-uracil-N-glycosylase carryover prevention into the combined assay. Restriction of as much as 0.5 microliter of deoxyuridine-containing amplification products has been achieved. The performance of the improved combined assay was compared directly with conventional nested RT-PCR and antibody detection. The combined assay was found to have sensitivity similar to that of nested RT-PCR in detecting HCV RNA from HCV antibody-positive specimens. In an analysis of hepatitis B virus antibody-positive specimens, nested amplification had false-positive rates ranging from 8 to 31%, while no false-positive results were seen with the combined assay. In comparison with serological methods, the combined assay had specificity and sensitivity of 100 and 95%, respectively.

Until HCV NS3/4A protease inhibitors become available at the end of 2011, the combination pegylated-interferon (PEG-IFN)-alfa and ribavirin (RBV) will remain the standard treatment for chronic hepatitis C patients. In some hepatitis C virus-infected patients, PEG-IFN plus RBV treatment against HCV should continue to be used because of side effects of new drugs such as anemia. Our Japanese experiences should provide new information for the treatment of chronic hepatitis C.

Quantitative serology for hepatitis B surface antigen (HBsAg) is a new candidate marker for prediction of clinical outcome. The aim of this study was to investigate the clinical significance of quantifying HBsAg in patients with hepatitis B virus (HBV) infection. A total of 424 patients who tested positive for HBsAg and were referred to Chiba University Hospital between January 1985 and April 2008 were included in the study, and the following characteristics were analyzed: age, gender, status of hepatitis B e antigen (HBeAg), alanine aminotransferase level (ALT), HBV DNA level, number of platelets and development of hepatocellular carcinoma. Measurement of HBsAg was performed using the chemiluminescent enzyme immunoassay method. The study group consisted of 239 men and 185 women, and their average age was 40.6 ± 14.0 years. HBsAg showed a positive correlation with HBV DNA level (Pearson's product moment correlation, r = 0.586, P0.001) and a weak inverse correlation with age (r = 0.3325, P0.001). A control study, matched with age and sex, was performed between two groups with and without HBeAg seroconversion during follow-up period. Compared with the age and sex-matched controls, the change in HBsAg levels per year showed a significant decrease 2 years before seroconversion (paired t-test, P0.05). The serial measurement of quantitative HBsAg level has the possibility of predicting the occurrence of HBeAg seroconversion.

Hepatocellular carcinoma has a decisive influence on the prognosis of cirrhotic patients, and the use of imaging modalities is essential for the screening, diagnosis and treatment of hepatocellular carcinoma. Ultrasound plays a major role among them, because it provides real-time and non-invasive observation by a simple and easy technique. In addition, ultrasound-guided needle puncture methods are frequently required for the diagnosis and/or treatment process of hepatocellular carcinoma. The development of digital technology has led to the detectability of blood flow by color Doppler ultrasound, and the sensitivity for tumor vascularity has shown remarkable improvement with the introduction of microbubble contrast agents. Moreover, near real-time three-dimensional ultrasound images are now available. These advancements in the ultrasound field have led to rapid progress in hepatocellular carcinoma management, and continuing advances are expected. This article reviews the current application of contrast-enhanced ultrasound for hepatocellular carcinoma in clinical practice.

There have been few reports that compare the effect of histamine H2-receptor antagonists (H2RAs) and proton pump inhibitors (PPIs) using continuous gastric pH monitoring for a long duration.To assess the early effects of both drugs on gastric pH using a wireless pH monitoring system.The test was conducted by a cross-over test: 10 healthy male volunteers were administered famotidine 20 mg twice a day and lansoprazole 30 mg once a day. Monitoring of gastric pH over four consecutive days was performed using a unique method we have developed that is an elaboration of the Bravo system.The time to reach a pH level of 3 or more with famotidine was significantly shorter than that for lansoprazole. The pH3 holding time ratio of famotidine during the first 4 h of administration was significantly higher than that of lansoprazole. The pH3 holding time ratio on each day from day 1 to day 3 was significantly higher following lansoprazole administration compared with famotidine administration.Famotidine was shown to act quickly in elevating gastric pH; however, lansoprazole was superior to famotidine in terms of its ability to elevate gastric pH for a long duration.

The recent development of the sensitive reverse transcription-polymerase chain reaction (RT-PCR) method allowed us to detect the presence of hepatitis A virus (HAV) RNA in sera from hepatitis A patients. To determine whether differences in HAV are related to the wide range of clinical severity, we used PCR-single strand conformation polymorphism (SSCP) to analyze the amplified product of the 5'-nontranslated region (5'NTR), where relatively homologous sequences are reported and the internal ribosomal entry site is considered to exist, from these various levels of hepatitis A. Twenty-seven patients admitted to Chiba University Hospital (between 1988 and 1997) were examined for HAV RNA in their sera by RT-PCR with primers located at 5'NTR of HAV RNA. The nucleotide (nt) sequence of a central part (nt 277-551) of 5'NTR was amplified and the product was examined by PCR-SSCP. HAV RNA was detected in all 27 hepatitis A patients examined: in 3 with fulminant hepatitis, 2 with severe acute hepatitis, and 22 with self-limited acute hepatitits. The amplified sequence revealed by PCR-SSCP showed that samples from fulminant and severe acute hepatitis patients had similar mobility in the gel, whereas that from acute hepatitis patients demonstrated a considerable variety in mobility patterns. From the analysis of the amplified product of 5'NTR of HAV, the bands are likely to be similar in the more severe forms of hepatitis A compared to those from uncomplicated self-limited acute hepatitis.

Adequate dosing of interferon (IFN) and its cost-effectiveness for sustained virological response were evaluated in relation to viral load and subtype. Prospective analysis of IFN therapy on 326 patients with chronic hepatitis C free from cirrhosis was performed using 9 or 6 million unit (MU) of IFN for six months daily and/or three times a week. Sustained virological response was achieved in 50-94% of patients withor =2 x 10(4) copies/ml (competitive RT-PCR) or100 x 10(3) copies/ml (Amplicor monitor) of HCV RNA by 468-1206 MU of IFN, but response was only 0-25% of the patients withor =2 x 10(5.5) copies/ml (competitive RT-PCR) or200 x 10(3) copies/ml (Amplicor monitor), even with 468-1206 MU of IFN. A high sustained rate was demonstrated in patients with 100-200 x 10(3) copies/ml of HCV RNA by 901-1206 MU of IFN, in comparison to that withor =900 MU of IFN. Multivariate analysis showed that IFN dose had a significant value for the efficacy of IFN therapy in patients presenting 100-200 x 10(3) copies/ml of HCV RNA. Cost efficacy analysis indicated that it cost approximately $10,000, $26,000, and $50,000-227,000 for one person-viral eradication in the patients with100, 100-200, and200 x 10(3) copies/ml, respectively. High-dose IFN is only cost effective in patients with intermediate viral loads, and IFN therapy could be recommended in patients with200 x 10(3) copies/ml of HCV RNA.

Previous studies on the effect of interferon therapy on the incidence of hepatocellular carcinoma have not sufficiently assessed degree of liver fibrosis, a major risk factor for hepatocellular carcinoma.To evaluate the effect of interferon therapy on incidence of hepatocellular carcinoma, adjusting for risk factors, including the degree of liver fibrosis.Retrospective cohort study.Seven university hospitals and one regional core hospital in Japan.2890 patients with chronic hepatitis C who had undergone liver biopsy since 1986. Of these patients, 2400 received interferon and 490 were untreated.The degree of liver fibrosis was assessed from stage F0 (no fibrosis) to stage F4 (cirrhosis). Response to interferon was determined virologically and biochemically. Screening for development of hepatocellular carcinoma was performed periodically during an average follow-up of 4.3 years. Effect of interferon therapy on the risk for hepatocellular carcinoma was analyzed by using Cox proportional hazards regression.Hepatocellular carcinoma developed in 89 interferon-treated patients and in 59 untreated patients. Among untreated patients, the annual incidence of hepatocellular carcinoma increased with the degree of liver fibrosis, from 0.5% among patients with stage F0 or F1 fibrosis to 7.9% among patients with stage F4 fibrosis. The cumulative incidence in treated and untreated patients differed significantly for patients with stage F2 fibrosis (P = 0.0128) and for those with stage F3 fibrosis (P = 0.0011). In multivariate analysis, interferon therapy was associated with a reduced risk for hepatocellular carcinoma (adjusted risk ratio, 0.516 [95% CI, 0.358 to 0.742]; P0.001), especially among patients with sustained virologic response (risk ratio, 0.197 [CI, 0.099 to 0.392]), among those with persistently normal serum alanine aminotransferase levels (risk ratio, 0.197 [CI, 0.104 to 0.375]), and among those with alanine aminotransferase levels less than two times the upper limit of normal (risk ratio, 0.358 [CI, 0.206 to 0.622]).Interferon therapy significantly reducesthe risk for hepatocellular carcinoma, especially among virologic or biochemical responders.

Recently, a novel hepatitis related virus was described and named as TTV. The positive rates of TTV DNA in healthy subjects to far reported ranged from 1% to 62%, therefore, there must be large differences in the prevalence of TTV in healthy subjects from country to country. However, detailed data are lacking in many reports, care should be taken in analyzing them. Generally, elderly subjects tend to have increased positivity in the same area. Sanitary environment may influence the positive rate. In addition the distribution of genotype may influence the positive rate. Although the high prevalence of TTV in healthy subjects may indicate the TTV as non-hepatopathogenic virus, further studies will be needed before drawing the conclusion.

It has been clarified that hepatitis G/GB virus-C is the minor cause of acute and chronic non-A-E hepatitis. But there exist non-A-E viral hepatitis patients. Recently, one of non-A-E hepatitis associated virus was identified and the new virus was named TT virus. We highly detected TT virus in the serum of hepatitis patients. TT virus were reported to be detected 1-37% of the general population in the world. TT virus may account for only a minor part of acute non-A-E hepatitis in Japan. However, whether TT virus infection really causes severe acute hepatitis is required to be elucidated.

Hepatitis C virus (HCV)-RNA status and alanine aminotransferase (ALT) levels determined shortly after interferon (IFN) therapy in patients with chronic hepatitis C do not predict long-term response. To determine the virological sustained response after the completion of IFN therapy, HCV-RNA was measured at the end of treatment and at 3-4 months and 12 months after the completion of therapy in 537 patients with chronic hepatitis C. In 347 patients, HCV-RNA was not detected by polymerase chain reaction (PCR) at the completion of therapy and 175 of these patients (50%) were still PCR negative 12 months later. In contrast, of the 180 patients who were HCV-RNA negative at 3-4 months after completion of therapy, 99% remained negative at 12 months. Normal ALT levels were found in 80, 93 and 95% of patients who were negative for HCV-RNA either at the end of treatment or at 3-4 months and 12 months after the completion of therapy, respectively. Of patients who were HCV-RNA positive, 30, 15 and 20% were found to have normal ALT levels at the same respective time points. To determine a sustained virological response shortly after the completion of therapy, serum HCV-RNA was serially examined in 66 patients negative for HCV-RNA at the end of therapy. Of 31 patients who relapsed, HCV-RNA reappeared in 33, 80, 97 and 100% of patients by 1, 2, 4 and 8 weeks after the completion of therapy. In conclusion, a sustained virological response could be determined with 97 and 99% certainty at 4 weeks and at 3-4 months after the completion of therapy, respectively.

During the course of hepatitis B virus (HBV) infection, there exists a long period of normal liver function tests with different states of HBeAg/Ab. As the state of HBV in asymptomatic carriers was not well characterized, we quantitatively and qualitatively examined HBV in both HBeAg-positive and anti-HBe-positive asymptomatic carriers. Sera from 10 HBeAg-positive and 27 anti-HBe-positive asymptomatic carriers were analyzed. The amount of HBV DNA was determined by dot-blot hybridization and polymerase chain reaction. The mutations in precore and core regions, spanning 636 nucleotides, of hepatitis B virus were examined by directly sequencing the amplified HBV DNA. HBV DNA was detected in all 10 HBeAg-positive cases, whereas it was found in only 7 of 27 (26%) anti-HBe-positive cases by the nested PCR method. The mean amount of HBV DNA in HBeAg-positive cases was 10(9.1 +/- 0.7) copies/ml, while that in anti-HBe-positive cases was 10(1.0 +/- 1.5) copies/ml. There were no missense mutations in the entire precore and core genes of HBV DNA taken from HBeAg-positive asymptomatic carriers. In contrast, many mutations (mean 9.0 +/- 3.3, range 6-14) were detected in the core gene of seven anti-HBe-positive asymptomatic carriers including two cases with increments of the mutations. Analysis of the precore region revealed three wild-type and four mutant-type (including one coexisting with wild-type) cases. These data suggest that HBV exists in quite different ways in "asymptomatic" carriers; in the HBeAg-positive phase HBV probably coexists with the host and remains as the wild type, whereas in the anti-HBe-positive phase a drastically reduced amount of HBV with many mutations remains, probably as a consequence of the long-lasting interaction with the host. Nevertheless, such small amount of virus could cause fulminant hepatic failure. It is important to make further clinical and virological investigations in order to understand the state of asymptomatic carrier.

The p53 gene has been shown to be commonly mutated in various human cancers, and mutant p53 can act as a dominant oncogene. The intact p53 protein is also known to induce the cyclin-dependent kinase inhibitor p21WAF1/CIP1 and is implicated in cell cycle arrest. We investigated p53 gene alterations in gastric adenocarcinoma and esophageal squamous cell carcinoma to elucidate the association of the nuclear accumulation of the p53 protein and/or p21WAF1/CIP1 protein. Abnormalities of the tumor suppressor gene p53 protein and the expression of p21WAF1/CIP1 protein were analyzed by immunohistochemical techniques in 32 cases of gastric adenocarcinoma and 15 cases of esophageal squamous cell carcinoma. Twenty cases of gastric cancer and five cases of esophageal cancer were also analyzed for p53 gene mutation by polymerase chain reaction and direct nucleotide sequencing. Overexpression of p53 protein was found in 13/32 (41%) of gastric cancers and 5/15 (33%) of esophageal cancers. We found immunodetectable p53 in 10/14 cases with mutations and in none of 11 cases without mutations in gastric and esophageal cancers. Hence, immunohistochemical and genetic analyses gave concordant results in 84% of 25 cases, revealing a good correlation between immunostaining of p53 and missense mutation of the p53 gene. p53 immunostaining was not observed in cases with frameshift or splicing mutation. The expression of p21WAF1/CIP1 protein was found in 9/32 (29%) of gastric cancers and 4/15 (27%) of esophageal cancers and in 2/14 (14%) cases with alteration of the p53 gene and in 5/11 (45%) without. These results suggest that abnormalities of p53 may be closely associated with the pathogenesis of gastric adenocarcinoma and esophageal squamous cell carcinoma and that the immunoreactivity of p53 protein is a general indicator of the tumors with altered p53 function. The expression of p21WAF1/CIP1 protein was suppressed in the neoplastic tissues with and without p53 gene alteration.

The relative role of virus load and hepatitis C virus (HCV) subtype as predictor of the efficacy of interferon (IFN) therapy is still in dispute. To resolve this issue, a multicenter, randomized, prospective study of 272 patients with chronic hepatitis C but without cirrhosis was conducted.The patients were randomly assigned to two different dose groups: 6 million units (MU) or 9 MU IFN three times a week for 6 months. Serum HCV RNA levels and HCV subtypes were determined.HCV RNA negativity rate at the completion of treatment with 9 MU IFN was higher than that with 6 MU (75% vs. 44%; P0.05). Virus eradication at 12 months after completion of treatment was higher in the 9 MU group than in the 6 MU group (36% vs. 25%; P0.05), especially in patients who had an intermediate virus load (10(4)-10(5) copies/mL by Amplicor monitor assay) (52% vs. 19%; P = 0.029). Virus eradication rate in patients with serotype 2 was higher than in those with serotype 1 for both regimens (6 MU, 53% vs. 15%; 9 MU, 76% vs. 29%; each P0.05).This prospective study showed that virus load, HCV serotype, and IFN dose are important predictors of the virological response to IFN therapy but virus load is the most important factor influencing the efficacy of IFN.

Recently, one of non-A to E hepatitis viruses was identified and the new virus was named HGV/GBV-C(HGBV-C). In Japan, more than 90% of liver cirrhosis and hepatocellular carcinoma were associated with hepatitis B or C virus infection. Proportion of HGBV-C infection was only 3.4% of patients with non-A to E chronic liver disease we examined. Thus, HGBV-C was considered to play only a minor role in chronic liver disease of unknown etiology. HGBV-C positive cases have a history of blood transfusion and/or operation and/or drug self-injection, and frequently co-infected with hepatitis C virus. Whether HGBV-C infection really causes severe chronic liver disease is required to be elucidated.

The effect of ursodeoxycholic acid on liver function tests and on bile acid metabolism was investigated in a multi-center randomized controlled dose study for chronic hepatitis C. Twenty, 18 and 19 patients were administered 150, 600 and 900 mg/day, respectively of ursodeoxycholic acid every day for 16 wk. Serum liver parameters and bile acid composition in the treatment groups were compared with 17 control patients. A similarly significant decrease of serum alanine aminotransferase and serum gamma-glutamyltransferase was observed in patients administered 600 and 900 mg of ursodeoxycholic acid. Serum bile acid composition was determined by high-performance liquid chromatography. At entry, the relative proportions of major bile acids were similar to those observed in normal individuals. Maximal concentrations of total ursodeoxycholic acid were 0.30 mumol/L, 5.59 mumol/L, 21.42 mumol/L and 14.73 mumol/L in the control, 150, 600 and 900 mg/day groups, respectively. The fraction of the total ursodeoxycholic acid increased in a dose-dependent manner, and it was significantly higher than in controls (p0.001). The hydrophobicity index of bile acids was calculated by the method of Heuman, and its correlation with serum parameter levels was analyzed. In the 600 and 900 mg/day dose groups, serum alanine aminotransferase decreased in the cases in which hydrophobicity index significantly decreased during treatment. The same correlation was observed between the hydrophobicity index and serum gamma = glutamyltransferase in these two groups. There was no correlation between these parameters in the control and 150-mg groups. There was no correlation between reduction rate of serum alanine aminotransferase and initial liver histology.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of interferon on chronic hepatitis C long after the treatments were assessed biochemically, virologically and histologically. Among the 12 cases who showed normal alanine aminotransferase (ALT) levels and seroconversion of HCV RNA after treatment, histological improvements were observed in 8. In contrast, among the 11 cases who showed abnormal ALT levels and HCV viremia after treatment, histological improvements were observed in only 1. Histological improvement of the liver was thus, well correlated with normalization of ALT levels and seroconversion of HCV RNA. These data indicate that complete histological resolution of chronic hepatitis C is obtainable by elimination of the virus by interferon treatment.

Two hundred thirteen patients with chronic hepatitis C were treated with IFN for 4 to 52 weeks and complete response, in which ALT levels sustained within the normal value after more than 6 months following IFN treatment, was achieved in only 10% of the patients treated with less than 300 MU of IFN, whereas it was 42% in those treated with more than 500 MU, hence the total amount of IFN is important in the treatment of chronic hepatitis C. The duration of the treatment (4 to 52 weeks) made little difference in the response when more than 500 MU of IFN were administered.

A 48-year-old woman with malignant lymphoma, who was diagnosed as a hepatitis B virus (HBV) carrier because of positive HBs antigen, was admitted to Chiba Cancer Center Hospital. Complete remission was achieved by two courses of combination chemotherapy, but fulminant hepatitis developed after withdrawal of chemotherapy and she died of hepatic failure. Retrospective analysis of viral DNA disclosed a mutation in the precore region of HBV-DNA in her serum. We consider that the precoremutant virus was associated with the pathogenesis of severe hepatic failure after withdrawal of cytotoxic chemotherapy.

Infection with hepatitis B virus leads to a wide spectrum of liver injury, including self-limited acute hepatitis, fulminant hepatitis and chronic hepatitis with progression to cirrhosis or acute exacerbation to liver failure, as well as, an asymptomatic chronic carrier state. To investigate why an extreme immunological attack occurs in fulminant hepatitis and severe exacerbation, the entire precore and core region were sequenced in 13 subjects. No significant change was noted in the 6 self-limited acute hepatitis patients. In contrast, clustering changes in a small segment of core codons #84 to #99 were found in all 7 fulminant and severe exacerbation patients. These data suggest that this mutation clustering region may play an important role in the pathogenesis, and such mutations appear to be related to severe liver damage.

We have treated 16 patients with chronic non-A, non-B hepatitis with short term (7-13 weeks) and long term (1 year) interferon. Sustained effect (normalization) of treatment was noted in all by long term treatment, where 7 of 8 with short term treatment showed flare-ups of transaminase after the therapy. In addition, negative turn of anti-HCV antibody was noted in 2 of 5 patients with long term treatment, whereas such change was noted in none of 6 anti-HCV antibody positive patients treated with short term treatment. These data suggest that long term interferon therapy may change natural course of chronic non-A, non-B hepatitis caused by HCV.

The presence of the enzyme bilirubin oxidase, which degrades bilirubin in vitro, was demonstrated in the liver. Subcellular-fractionation experiments indicate that bilirubin oxidase is located in both the inner and outer membranes of the mitochondria. The mean rate of the reaction is 1.57 +/- 0.38 (S.D.) nmol of bilirubin degraded/min per mg of mitochondrial protein (munits/mg of protein). With respect to the overall breakdown of bilirubin, the enzyme has a Km' of 136 microM-bilirubin and a Vmax.' of 9.13 munits/mg of protein. Its activity is influenced by the ionic strength of the media and is inhibited by KCN, thiol reagents, NADH and albumin. The enzyme is aerobic, and between 1 and 1.5 mol of O2 are consumed per mol of bilirubin degraded. The products of the reaction include propentdyopents. The hepatic bilirubin oxidase activity of the jaundiced Gunn-rat liver is not significantly different from that of the Sprague-Dawley rat, and it is not induced by beta-naphthoflavone.