To estimate the prevalence of hepatitis B virus(HBV) and hepatitis C virus(HCV) infection and to determine associated risk factors, a population-based seroepidemiologic study was carried out. In 1993, a health examination survey of the population was carried out in rural area known to have a high incidence of liver cancer. The study population were those who volunteered to participate in a health survey over 10 years of age. Examinees were interviewed by specially trained staffs. Sera from 1,033 study subjects were tested for hepatitis B surface antigen (HBsAg) by reverse passive hemagglutinin (RPHA) estimation and for hepatitis C virus antibody (anti-HCV) by 2nd generation passive hemagglutinin (PHA) estimation, The age and sex standardized prevalence of HBsAg was 6.3% which was similar to national average, but that of anti-HCV was 5.1% which was 4 to 5 times higher than that of blood or other health examinees in Korea. In a multivariate analysis, transfusion history, surgical operative history, and acupuncture history were not associated with HBsAg positivity. In contrast, acupuncture history (adjusted odds ratio[OR]=2.2 : 95% Confidence interval[CI] 1.0-4.7) and surgical operative history(adjusted OR=2.0 : 95% CI 1.0-4.1) were associated with anti-HCV positivity. The present study suggest that there is an highly endemic area of HCV infection in Korea and probably this endemicity is associated with a parenteral source of HCV infection other than blood transfusion.
Acupuncture ; Blood Transfusion ; Health Surveys ; Hemagglutinins ; Hepacivirus ; Hepatitis B Surface Antigens ; Hepatitis B* ; Hepatitis C* ; Hepatitis* ; Incidence ; Korea* ; Liver Neoplasms ; Multivariate Analysis ; Prevalence ; Risk Factors ; Seroepidemiologic Studies*

In order to optimize the fabrication of SiO2 tubes immobilized with antibody for hepatitis C virus antigen (HCAg) detection, we formed the activated amino on the surface of SiO2 tubes by using the activation of aminosilane. Then we immobilized the hepatitis C virus (HCV) monoclonal antibody on the surface of SiO2 tubes by using glutaraldehyde as a chemical cross-linker, followed by detecting HCAg. Sequence tests showed that when the SiO2 tubes were treated in 10% (V/V) aminosilane solution and 3% (V/V) glutaraldehyde solution for 3 hours and 2 hours, respectively, the HCV monoclonal antibody had high immobilization efficiency and low nonspecificity, and the HCAg was detected to 1 ng/mL. This experiment can provide principle and experimental data for establishment of HCAg magnetic immunoassay system.
Antibodies, Immobilized ; immunology ; Antibodies, Monoclonal ; chemistry ; immunology ; Hepatitis C Antibodies ; chemistry ; immunology ; Hepatitis C Antigens ; analysis ; immunology ; Humans ; Silicon Dioxide ; chemistry

BACKGROUND/AIMS: As a preliminary study to test the possibility of oral transmission of hepatitis C virus (HCV), many investigations in order to detect the extrahepatic localization of HCV have been performed. In this study, we examined the presence of HCV viral proteins in gastric mucosa. METHODS: Immunohistochemical staining to NS3 protein were done to detect the HCV virus in gastric mucosa. The results were compared with NS5a protein staining to confirm the NS3 protein staining. RESULTS: Total of 164 patient were included. 58 patients with anti-HCV (+) were designated to case group and 70 with anti-HCV (-) to control group. 36 were excluded in this study due to concomittent illness. Anti-HCV (+) group showed 50.0% (29/58) of positivity to NS3 protein staining and anti-HCV (-) group showed 12.6% (9/70) of positivity (p<0.001). Immunohistochemical staining to NS5a protein were done to validate the result of NS3 (+) staining in the anti-HCV (+) group (n=58). NS5a (+) staining were observed in 58.6% (34/58). The results of NS5a staining were consistent with that of NS3 in 70.7%. The reliability coefficients by Chronbach's Alpha for NS3 and NS5a stain test was 0.59. CONCLUSIONS: HCV can exist in gastric mucosal cell as an extrahepatic presence. In the future, this study may provide some fundamental data for the research of possible oral transmission route of HCV.
Aged ; English Abstract ; Female ; Gastric Mucosa/*virology ; Hepacivirus/*isolation & purification ; Hepatitis C Antigens/*analysis ; Humans ; Male ; Middle Aged

OBJECTIVESTo identify hepatic progenitor cells (HPCs) in patients with severe hepatitis (SH) by detecting their markers and investigate the features of their distribution and location.

METHODSLiver tissues taken from 59 SH patients were tested for the receptor of stem cell factor (c-kit), pi-class glutathione S-transferase (GST-pi), cluster of differentiation 34 (CD34), cytokeratin 19 (CK19), cytokeratin 18 (CK18) and alpha fetoprotein (AFP) by immunohistochemistry (IHC). Meanwhile, 58 patients with acute or chronic hepatitis were also detected to act as controls.

RESULTSHepatic progenitor cells could be seen in SH patients. Most of them existed as ductular cells that had been called "typical ductular proliferation (ADP)" or "typical ductular reaction" in previous research. These ductular cells were mainly located at the portal areas, fibro septa, periportal parenchyma and the border of the pseudolobuli and inflammatory foci. Further, c-kit, GST-pi, CK19 and CK18, but not CD34 and AFP could be detected in these cells. Another kind of HPC was the small hepatocyte-like cell (SHLC), which could express c-kit, GST-pi, and CK18, but not CK19, CD34 and AFP. The semi-quantitative analysis showed that the scope of ADP in SH patients was significantly larger than that in acute and chronic hepatitis patients (chi2= 63.62, P<0.05), and the scope of ADP in subacute severe hepatitis and chronic severe hepatitis patients was also significantly larger than that in acute severe hepatitis patients.

CONCLUSIONIn the course of regeneration of viral hepatitis, different types of pathology have different features. In acute and chronic hepatitis (G1-2), the regeneration is mainly owing to the proliferation of mature hepatocytes, and in chronic hepatitis (G3-4), there is the participation of HPCs, although they are limited. In severe hepatitis, however, since the replicative capacity of normal hepatocytes is impaired or prohibited, liver regenerates and restores mainly by the means of hepatic stem cells activation and proliferation. But the hepatic stem cells don't differentiate into their mature functional compartments directly at all. There are several intermediary or transition populations. In human severe hepatitis, they are mainly ductular cells, and parts of them are small hepatocyte-like cells.

Antigens, CD34 ; analysis ; Cell Division ; Female ; Glutathione S-Transferase pi ; Glutathione Transferase ; analysis ; Hepatitis ; pathology ; Hepatocytes ; pathology ; Humans ; Immunohistochemistry ; Isoenzymes ; analysis ; Keratins ; analysis ; Male ; Proto-Oncogene Proteins c-kit ; analysis ; Stem Cells ; pathology ; alpha-Fetoproteins ; analysis

The prevalence of hepatocellular carcinoma (HCC) has increased in recent years. However, HCC remains poorly characterized in elderly patients, and comprehensive data are limited. This study aimed to investigate the clinical characteristics, prognostic features and survival outcome of elderly HCC patients. We retrospectively analyzed 992 HCC patients treated at Dongsan Hospital from January 2003 to December 2007. The patients were divided into two age groups: < 70 yr (n = 813) and > or = 70 yr (n = 179). Elderly HCC patients, compared to younger patients, had significantly higher incidence of females (31.3% vs 18.9%, P = 0.001), hepatitis C-related disease (HCV antibody positivity 26.3% vs 9.2%, P = 0.001) and comorbid condition (53.6% vs 32.1%), but lower rates of hepatitis B-related disease (HBs antigen positivity 31.3% vs 69.4%, P = 0.001). There were no significant differences in underlying liver function, stage and survival outcomes. Factors significantly influencing the prognosis of HCC were Child-Pugh grade, number of HCC, level of alpha-fetoprotein, presence of metastasis. The survival outcome of older patients with HCC was not different from that of younger patients. There were no differences between groups in independent factors influencing the prognosis of HCC. Therefore, determining the optimal management strategy for elderly HCC patients is important to improve survival and long-term outcomes.
Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; Carcinoma, Hepatocellular/etiology/*mortality/*therapy ; Chemoembolization, Therapeutic ; Female ; Hepatitis B/complications/diagnosis ; Hepatitis B Surface Antigens/blood ; Hepatitis C/complications/diagnosis ; Hepatitis C Antibodies/blood ; Humans ; Liver Neoplasms/etiology/*mortality/*therapy ; Male ; Middle Aged ; Palliative Care ; Regression Analysis ; Republic of Korea ; Retrospective Studies ; Survival Analysis ; alpha-Fetoproteins/analysis

OBJECTIVETo establish the method to detect the cellular immune response of enhanced hepatitis B vaccine and make verification preliminary.

METHODSImmunized BALB/c mice with enhanced hepatitis B vaccine and detected the IFN-gamma spots forming cells (SFC) of mouse spleen cell by Elispot. Optimized the conditions of the experiment. Cellular immune response between enhanced hepatitis B vaccine and normal hepatitis B vaccine by Elispot were compared.

RESULTSIFN-gamma SFC was higher in 5microg dose than in 2microg dose after immunization with enhanced hepatitis B vaccine and IFN-gamma SFC was declined after immunization 3 weeks ago. IFN-gamma SFC was higher in stimulus by peptide than by protein. Compared to normal hepatitis B vaccine, IFN-gamma SFC was higher in enhanced hepatitis B vaccine.

CONCLUSIONEstablished the detection method to evaluate the cellular immunity of enhanced hepatitis B vaccine and tested the repeatability.

Animals ; Enzyme-Linked Immunospot Assay ; methods ; Female ; Hepatitis B ; immunology ; prevention & control ; virology ; Hepatitis B Surface Antigens ; administration & dosage ; immunology ; Hepatitis B Vaccines ; administration & dosage ; immunology ; Humans ; Immunity, Cellular ; Interferon-gamma ; analysis ; immunology ; Mice ; Mice, Inbred BALB C

OBJECTIVETo investigate the correlation between liver stem cell activation and histopathologic changes in liver transplant recipients with hepatic failure.

METHODSA total of 33 cases of hepatic failure were enrolled into the study. Donor liver tissues were used as normal controls. Histopathologic changes, presence of hepatotropic virus antigens, history of artificial liver therapy and number of c-kit positive cells were analyzed.

RESULTSThere were a total of 25 males and 8 females. The age of patients ranged from 21 to 64 years. Among the 33 patients studied, 6 suffered from acute liver failure, 5 from subacute liver failure and the remaining from acute-on-chronic liver failure (associated with cirrhosis). Thirteen patients had a history of artificial liver therapy. Activated liver stem cells expressed c-kit monoclonal antibody but were negative for toluidine blue stain. The number of c-kit-positive cells in acute liver failure, subacute liver failure and acute-on-chronic liver failure were 3.50 +/- 2.66 (0 to 8) per mm(2), 11.47 +/- 8.85 (3 to 30) per mm(2) and 15.50 +/- 10.95 (5 to 45) per mm(2), respectively (P < 0.05). The number of c-kit-positive cells in cases with or without artificial liver therapy showed no statistically significant difference.

CONCLUSIONSThe poor prognosis of acute liver failure is mainly due to massive liver necrosis and insufficient stem cell activation. Liver stem cell level is increased whenever there is progression into subacute liver failure and chronic liver failure. Actively treating acute liver failure with stimulation of the self-regeneration system in liver is thus useful.

Adult ; Female ; Hepatitis B ; Hepatitis B Surface Antigens ; analysis ; Hepatitis B e Antigens ; analysis ; Hepatitis B virus ; isolation & purification ; Humans ; Ki-67 Antigen ; metabolism ; Liver Failure ; metabolism ; pathology ; virology ; Liver Failure, Acute ; metabolism ; pathology ; virology ; Liver, Artificial ; Male ; Middle Aged ; Proto-Oncogene Proteins c-kit ; metabolism ; Stem Cells ; metabolism ; Young Adult

BACKGROUNDRNA interference (RNAi) is a powerful tool to silence gene expression post-transcriptionally. Our previous study has demonstrated that small interfering RNAs (siRNAs) have sufficiently inhibited hepatitis B virus (HBV) replication and expression in vitro. In this study we observed the RNAi-mediated inhibitory effects on HBV replication in mice models and accessed the specificity of these effects.

METHODSA mutant RNAi vector (pSI-C mut) with two base pairs different from the original target gene sequence at the RNAi vector (pSI-C) was constructed according to the method described in this study. A mouse model of acute hepatitis B virus infection was established by injecting naked plasmid pHBV1.3 via the tail vein with acute circulatory overload. pSI-C, pSI-C mut and the irrelevant RNAi control plasmid for green fluorescent protein (GFP) gene, pSIGFP were respectively delivered with pHBV1.3 by tail vein injection method. Six days post injection, enzyme-linked immunosorbent assay (ELISA) assay was used to measure the concentration of HBV surface antigen (HBsAg) in mouse serum, immunohistochemical straining method was used to visualize the expression of HBV core protein (HBcAg) in liver tissues, and the transcriptional level of HBV C mRNA in liver tissues was detected by reverse transcriptase PCR (RT-PCR) analysis.

RESULTSInjection of pSI-C exerted magnificent and specific inhibitory effects on the replication and expression of HBV in the murine model. After 6-day post-injection (p.i.), the OD values were shown to be 5.07 +/- 1.07 in infecting group and 0.62 +/- 0.59 in pSI-C group. The concentration of HBsAg in pSI-C group was significantly lower than that in infecting group (P < 0.01). Liver intracellular synthesis of viral core protein was sharply reduced to 0.9% +/- 0.1%, compared with 5.4% +/- 1.2% of positive hepatocytes in infecting group (P < 0.01), and the transcriptional level of HBV C mRNA was greatly reduced by 84.7%. However, the irrelevant RNAi control plasmid (pSIGFP), and the pSI-C mut did not show the same robust inhibitory effects as pSI-C.

CONCLUSIONpSI-C exert efficient and specific inhibitory effects on HBV replication and expression in mice models.

Animals ; Hepatitis B ; therapy ; virology ; Hepatitis B Core Antigens ; biosynthesis ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; physiology ; Male ; Mice ; Mice, Inbred BALB C ; RNA Interference ; RNA, Messenger ; analysis ; RNA, Small Interfering ; therapeutic use ; RNA, Viral ; analysis ; Virus Replication

BACKGROUND/AIMS: Alcohol and the hepatitis B virus (HBV) exert synergistic effects in hepatocelluar carcinogenesis. We aimed to elucidate the clinical significance of the antibody to hepatitis B core antigen (anti-HBc) and occult HBV infection on the development of hepatocellular carcinoma (HCC) in patients with alcoholic liver cirrhosis (LC). METHODS: Patients with alcoholic LC alone (n=193) or combined with HCC (n=36), who did not have HBsAg or antibody to hepatitis C virus were enrolled. Clinical data and laboratory data including anti-HBc were investigated at enrollment. The polymerase chain reaction was applied to HBV DNA using sera of patients with HCC or LC after age and sex matching. RESULTS: Patients with HCC were older (60+/-11 years vs. 53+/-10 years, mean+/-SD, P<0.001), more likely to be male (100% vs. 89%, P=0.03), and had a higher positive rate of anti-HBc (91.2% vs. 77.3%, P=0.067), and a higher alcohol intake (739+/-448 kg vs. 603+/-409 kg, P=0.076) than those with LC. Age was the only significant risk factor for HCC revealed by multiple logistic regression analysis (odds ratio, 1.056; P=0.003). The positive rate of anti-HBc and alcohol intake did not differ in age- and sex-matched subjects between the LC (n=32) and HCC (n=31) groups. However, the detection rate of serum HBV DNA was higher in the HCC group (48.4%) than in the LC group (0%, P<0.001). CONCLUSIONS: Anti-HBc positivity is not a risk factor for HCC. However, occult HBV infection may be a risk factor for HCC in patients with alcoholic LC.
Adult ; Aged ; Antibodies, Viral/blood ; Carcinoma, Hepatocellular/diagnosis/epidemiology/*etiology ; DNA, Viral/analysis ; Female ; Hepatitis B/*complications/diagnosis ; Hepatitis B Core Antigens/*immunology ; Hepatitis B Surface Antigens/immunology ; Hepatitis B virus/genetics/immunology/isolation & purification ; Hepatitis C/complications/diagnosis ; Humans ; Liver Cirrhosis, Alcoholic/*complications/diagnosis/epidemiology ; Liver Neoplasms/diagnosis/epidemiology/*etiology ; Male ; Middle Aged ; Risk Factors

BACKGROUND/AIMS: We investigated the frequency of occult hepatitis B virus (HBV) infection in anti-hepatitis C virus (HCV)-positive individuals and the effects of occult HBV infection on the severity of liver disease. METHODS: Seventy-one hepatitis B virus surface-antigen (HBsAg)-negative patients were divided according to their HBV serological status into groups A (anti-HBc positive, anti-HBs negative; n=18), B (anti-HBc positive, anti-HBs positive; n=34), and C (anti-HBc negative, anti-HBs positive/negative; n=19), and by anti-HCV positivity (anti-HCV positive; n=32 vs. anti-HCV negative; n=39). Liver biopsy samples were taken, and HBV DNA was quantified by real-time PCR. RESULTS: Intrahepatic HBV DNA was detected in 32.4% (23/71) of the entire cohort, and HBV DNA levels were invariably low in the different groups. Occult HBV infection was detected more frequently in the anti-HBc-positive patients. Intrahepatic HBV DNA was detected in 28.1% (9/32) of the anti-HCV-positive and 35.9% (14/39) of the anti-HCV-negative subjects. The HCV genotype did not affect the detection rate of intrahepatic HBV DNA. In anti-HCV-positive cases, occult HBV infection did not affect liver disease severity. CONCLUSIONS: Low levels of intrahepatic HBV DNA were detected frequently in both HBsAg-negative and anti-HCV-positive cases. However, the frequency of occult HBV infection was not affected by the presence of hepatitis C, and occult HBV infection did not have a significant effect on the disease severity of hepatitis C.
Adult ; Aged ; Cohort Studies ; DNA, Viral/analysis ; Female ; Genotype ; Hepatitis B/*complications/*diagnosis ; Hepatitis B Core Antigens/blood/immunology ; Hepatitis B Surface Antigens/blood/immunology ; Hepatitis B virus/*genetics ; Hepatitis C, Chronic/*complications/genetics/*pathology ; Humans ; Liver/virology ; Male ; Middle Aged ; Polymerase Chain Reaction ; Severity of Illness Index