Evaluation of antiviral efficacy of Chinese traditional medicine Babao Dan in rabbits infected with hepatitis E virus.

Hepatitis E virus (HEV) is a major cause of acute viral hepatitis. Patients with chronic hepatitis B superinfected with HEV may progress to liver failure. Babao Dan (BD) is a traditional Chinese medicine widely used as an auxiliary option for the treatment of chronic hepatitis and liver cancer in China. This study aimed to evaluate the effect of BD on the management of HEV infection in a rabbit model. Sixty-two specific-pathogen-free (SPF) rabbits were divided randomly into five groups and treated with BD or placebo for 2 weeks. All rabbits were inoculated intravenously with rabbit HEV after initial administration. Then, rabbits were administered BD or ribavirin or placebo at 2 weeks post-inoculation (wpi) until faecal virus shedding showed negative. The duration of faecal virus shedding and levels of HEV RNA in faeces were reduced, and anti-HEV antibodies were detected in all rabbits in groups treated with BD before or after inoculation. Ribavirin treatment rapidly cleared HEV infection in SPF rabbits, but anti-HEV antibodies remained negative in 50 % of rabbits treated with ribavirin. These results indicate that ribavirin treatment was more effective in clearing HEV infection, while administration of BD before or after inoculation was effective in clearing HEV infection. Further clinical studies are warranted.

Predicting acute viral hepatitis serum markers (A and E) in patients with suspected acute viral hepatitis attending primary health care centers in Baghdad: a one year cross-sectional study.

BACKGROUND: Viral hepatitis is an important preventable infectious disease with various rates of occurrence in different areas of the world. The main objective of the present study was to evaluate the role of some risk factors in predicting a positive acute viral hepatitis marker among patients with suspected acute viral hepatitis in a primary health care setting in Baghdad. Besides, finding out the occurrence of jaundice, contribution of viruses A and E to the cases that have occurred in Baghdad province was also searched for. METHODS: Over a period of 1 year a descriptive cross sectional study was carried out at the primary health care centers in Baghdad. A questionnaire form was used to collect data about demographic factors and the results of the investigations. Total serum bilirubin and bilirubin in urine were done at the primary health care center laboratory. The rest of the sera samples were sent to Hepatitis referral Lab at Central Public Health Laboratory (CPHL) to be tested for anti HAV IgM and anti HEV IgM using ELISA technique. RESULTS: A total of 7,576,372 consultations to primary health care centers were recorded in Baghdad. Among those a total of 2,692 cases (35.5 per 100,000 consultations) were labeled as acute viral hepatitis cases. A positive hepatitis viral marker (A, B, C and E) was found in 1,332 cases (17.6 per 100,000 consultations). More than two fifths (44.8%) of cases were positive for anti-HAV antibodies and another 1.6% had positive anti-HEV antibodies. CONCLUSION: During 1 year period, the rate of occurrence of suspected acute viral hepatitis cases was 35.5 per 100000 of consultations to the primary health care centers in Baghdad. Of the total suspected cases, only 17.6 per 100000 of the consultations were positive for one of the viral hepatitis markers. Those who tested positive for one of the viral hepatitis markers represent 49.5% of the suspected cases. Proportion of anti HAV IgM positive tests among suspected cases was 44.8%. Factors that were able to predict positive Anti HAV IgM were (younger age group, negative history of cupping or tattooing, negative past history of jaundice). Male gender was the least important predictor of a positive case for anti HAV IgM. Proportion of Anti HEV IgM positive tests among suspected cases was 1.6%. Of the factors studied, only age was able to predict a positive Anti HEV IgM in those more than 15 years. Further studies are recommended.

Infection dynamics of hepatitis E virus in naturally infected pigs in a Chinese farrow-to-finish farm.

To analyze the changes that occur in pigs during hepatitis E virus (HEV) infection, 256 serial serum samples were obtained from 32 pigs from one pig farm at ages 0 (cord blood), 15, 30, 60, 75, 90, 120, and 150 days. All HEV markers were assayed in these samples and showed that total anti-HEV antibodies and IgG formed two peaks. The first peak occurred at 0-60 days and the second after 75 days. No markers of infection, such as HEV RNA, antigen and anti-HEV IgM, were detectable during the first peak. Most newborn piglets (< 24 h of age) were negative for total anti-HEV and IgG. However, colostrum from all of the sows had evidence of these antibodies. Thus, the anti-HEV in the first peak was assumed to be acquired from maternal milk. Some infectious markers were positive at the beginning of second peak. PCR products were cloned and sequenced and the results indicated those sequences belonged to HEV genotype 4. The antibody present during the second peak may be induced by natural infection with HEV. In conclusion, pigs are susceptible to HEV infection and may remain infectious after the first peak of anti-HEV antibody.

Anti-hepatitis B virus activities of triterpenoid saponin compound from Potentilla anserine L.

The Tibetan herb Potentilla anserina L. has been widely used in China for many thousands of years to treat hepatitis-B. Bioassay-guided fractionation of the ethanol extract of the rhizomes led to the isolation of a triterpenoid saponin (TS) that was determined to be 2alpha,3beta,19alpha-trihydroxyurs-12-en-28-oic acid beta-D-glucopyranosyl ester. Using models of HBV infection, this compound was evaluated for its effect on HBV antigene expression in the 2.2.15 cell line in vitro and anti-hepatitis B virus (HBV) activities in Peking ducklings in vivo. Results showed that it could decrease the expression levels of HBsAg, HBeAg and HBVDNA in the 2.2.15 cell culture and the inhibitory effect was not due to the cytotoxity of the triterpenoid saponin. The antiviral study in vivo on Peking ducklings also demonstrated that this compound inhibits duck hepatitis B virus (DHBV) DNA replication.

An outbreak of hepatitis A associated with green onions.

Forty-three cases of serologically confirmed hepatitis A occurred among individuals who ate at restaurant A in Ohio in 1998. Serum samples from all restaurant A employees who worked during the exposure period were negative for IgM antibodies to hepatitis A virus (HAV). A matched case-control study determined that foods containing green onions, which were eaten by 38 (95%) of 40 case patients compared with 30 (50%) of 60 control subjects, were associated with illness (matched odds ratio, 12.7; 95% confidence interval, 2.6-60.8). Genetic sequences of viral isolates from 14 case patients were identical to each other and to those of viral isolates from 3 patients with cases of hepatitis A acquired in Mexico. Although the implicated green onions, which could have come from one of 2 Mexican farms or from a Californian farm, were widely distributed, no additional green onion-associated cases were detected. More sensitive methods are needed to detect foodborne hepatitis A. A better understanding of how HAV might contaminate raw produce would aid in developing prevention strategies.

Interferon-alpha may exacerbate cryoblobulinemia-related ischemic manifestations: an adverse effect potentially related to its anti-angiogenic activity.

The discovery of the strong association between hepatitis C virus (HCV) infection and the development of mixed cryoglobulinemia has motivated active testing of antiviral-directed alternative therapies. Several trials have demonstrated that classic cryoglobulinemia-associated manifestations improve with interferon-alpha (IFNalpha) treatment. Herein we report on 3 HCV-infected patients with severe cryoglobulinemia-related ischemic manifestations who were closely followed up during IFNalpha therapy. Clinical evaluations with special attention to ischemic lesions, liver function tests, and cryocrit determinations were serially performed. In addition to prednisone and immunosuppressive agents, the patients received IFNalpha at 3 x 10(6) units, 3 times per week for 2 months, 3 months, and 4 months, respectively. In all 3 patients, systemic features improved, liver function results returned to normal, and cryocrit values decreased. However, ischemic lesions became less vascularized and ischemia progressed, leading to transmetatarsal and subcondylar amputation, respectively, in 2 of the patients and fingertip necrosis and ulcer enlargement in the third. Skin biopsies performed before IFNalpha therapy and after 2 months of IFNalpha therapy in the third patient showed a significant decrease in subepidermal microvessels. When IFNalpha was discontinued, the lesions finally healed. Cryoglobulinemia-related ischemic lesions may worsen during IFNalpha treatment, presumably through a decrease in inflammation-induced angiogenesis. The anti-angiogenic activity of IFNalpha may delay the appropriate healing of ischemic lesions.

Epidemiological and clinical aspects of hepatitis G virus infection in blood donors and immunocompromised recipients of HGV-contaminated blood.

BACKGROUND AND OBJECTIVES: The infectiousness and clinical relevance of the newly discovered blood-borne Flaviviridae-like agent, termed hepatitis G virus (HGV), are not well understood. MATERIALS AND METHODS: Twenty-three transfusion recipients of two HGV-affected long-term blood donors were studied for HGV genome and antibodies to the putative envelope 2 glycoprotein (anti-E2) of HGV. Nine recipients had nonhematological disorders and 14 suffered from severe hematological diseases and 7 of them received allogeneic bone marrow or blood stem cell transplantation. The molecular epidemiology of the observed HGV infection was studied by direct sequencing of parts of the 5'-noncoding region, NS3, and NS5 region of HGV in the 2 long-term donors and in their 6 recipients who became HGV RNA positive. Additionally, 549 individuals-homologous (n = 254) and autologous blood donors (n = 202), and medical staff (n = 89)--were investigated for the presence of HGV RNA. RESULTS: HGV RNA in serum was found in 15 of the 23 (65%) transfusion recipients with known exposure of HGV-contaminated blood. Seven of the remaining 8 recipients showed only an anti-E2 response, indicating previous HGV infection with spontaneous clearance of the virus. In one recipient neither HGV RNA nor anti-E2 could be detected. Molecular evidence for HGV transmission by the 2 donors was found in 3 of the 6 recipients studied. The alanine aminotransferase levels were not significantly different in the HGV RNA positive and negative recipients, and none of the 23 recipients developed posttransfusion hepatitis. Persistent HGV infection was observed especially in recipients with severe hematological disorders or in those in whom intensive immunosuppressive treatment was necessary. Of the 549 individuals studied, 10 (1.8%) were healthy carriers of HGV RNA. CONCLUSION: The persistence of transfusion-acquired HGV infection is not associated with acute or chronic hepatitis, but may be influenced by the recipient's underlying disease.

[Clinical study of TCM-WM on aplastic anemia complicated with hepatitis C].

The testing kit of second generation for serum anti-HCV was used in 82 cases of aplastic anemia (AA). The results showed that positive rate was 69.4% (43/62) in the patients of AA with transfusion, this was significantly higher than that in the patients of AA without transfusion. There was no difference of anti-HCV antibody positive rate between chronic AA and acute AA (P > 0.05), incidence rate of post-transfusion hepatitis C (PTHC) in AA was 33.9% (21/62), among which the incidence rate in acute and chronic AA were 68.8% (11/16) and 21.7% (10/46) respectively (P < 0.01). The anti-HCV positive patients were divided into two groups: PTHC and non-PTHC, there was no statistical difference of their transfusion volume, hemoglobin, white blood cell between these groups. Response rate of AA was lower in anti-HCV positive patients than that in negative patients (P < 0.05). Acute, icteric PTHC was predominant in patients with AA. The patients with AA complicated with PTHC was liable to bleed and be infected. PTHC has been an important complication in patients with AA. The better response was obtained by TCM-WM therapy in the patients.

[A trial of a hepatitis A cultured inactivated vaccine on rhesus macaques]. / Ispytanie kul'tural'noi inaktivirovannoi vaktsiny gepatita A na makakakh rezusakh.

In this work experimental model of hepatitis A virus (HAV) infection in macaques rhesus was used. In 6 seronegative monkeys immunized with the inactivated vaccine (3 injections of 0.3 micrograms of viral protein each at an interval of 1 month) pronounced antibody response was observed. The dynamics and titers of anti-HAV antibodies were similar to those in 5 rhesus macaques which received the active virus. But, in contrast to the latter, no IgM antibodies were detected in the immunized animals. Three months after the end of immunization the monkeys were resistant to challenge with a HAV strain pathogenic for humans (10(3) - 10(4) ID50). The monkeys had no morphological changes in the liver and no rise in the serum alanine-amino transferase activity, but exhibited transient excretion of the virus in feces, as well as stimulation of anti-HAV antibodies (in the absence of IgM antibodies). The vaccine under test proved to be safe, immunogenic and produced a protective effect.

The effect of drinking coffee and smoking cigarettes on the risk of cirrhosis associated with alcohol consumption. A case-control study. Provincial Group for the Study of Chronic Liver Disease.

In order to assess the interaction between alcohol intake, tobacco smoking and coffee consumption in determining the risk of liver cirrhosis we carried out a hospital-based case-control study involving 115 patients at their first diagnosis of cirrhosis and 167 control patients consecutively enrolled in the General Hospitals of the Province of L'Aquila (Central Italy). The mean life-time daily alcohol intake (as g ethanol consumed daily) was measured by direct patient interviews, whose reproducibility was > 0.80 and similar for cases and controls, as checked by interviewing the relatives of a sample of 50 cases and 73 controls. During the same patient's interview we also measured the mean consumption of coffee (daily number of cups of filtered coffee) and tobacco (life-time daily number of cigarettes smoked). A dose-effect relationship on the risk of cirrhosis was present both for alcohol intake--for which the risk was significantly increased above 100 g of daily intake--and for cigarette consumption. The latter did not however improve the goodness-of-fit of a logistic regression model including alcohol intake as covariate. By contrast, coffee consumption had a protective effect on the risk of cirrhosis and significantly improved the goodness-of-fit of such a model. Abstaining from coffee consumption determined both a significantly increased risk of cirrhosis, even for daily alcohol intake below 100 g, and a multiplicative effect with alcohol intake on this risk. In patients drinking > or = 101 g ethanol daily the relative risk increased from 5.5 (95% confidence interval: 1.4-22.0) for coffee consumers to 10.8 (95% confidence interval: 1.3-58.1) for coffee abstainers.(ABSTRACT TRUNCATED AT 250 WORDS)

[Efficacy of Phyllanthus spp. in treating patients with chronic hepatitis B].

The efficacy of Phyllanthus amarus produced in india, P. niruri gathered from hainan province and P. urinaria from henan province was assessed in a total of 88 cases of chronic hepatitis B with 11.42 and 35 each. It was shown that P. urinaria had the effect of seroconversion on HBeAg from positive to negative as well as on HBeAb from negative to positive, while the other two herbs had not. In addition none of these three herbs had similar effect on HBsAg.

[Prevalence of HIV and HCV among injecting drug users (IDUs) in Yunnan, China].

Prevalence of HIV and HCV was studied among 507 IDUs in Yunnan Province. The results reveal that sharing needles with other IDUs is one of the primary pathways to transmit both HIV and HCV. The rates of HIV and HCV infection in the IDU group is 66.5% and 94.9% respectively, which is significantly higher than those in the non-IDU group. There is a direct correlation between the rates of HIV and HCV antibody positive. 97.5% HCV (+) subjects are also HIV (+); and 57% HIV (+) subjects are HCV (+). It is suggested that all IDUs should be encouraged to stop using drugs to prevent HIV and HCV transmission.

Quantitative detection of hepatitis B virus DNA in sera from patients with acute hepatitis B.

Two hundred forty-four serial serum samples from 30 adults hospitalized with benign (nonfulminant) acute hepatitis B were tested for the presence of hepatitis B virus (HBV) DNA by a quantitative solution hybridization assay using a 125I-labeled DNA probe complementary to HBV-DNA sequences. Acute hepatitis B was self-limiting in 28 and progressed to chronicity in the remaining two patients. Of the 28 patients with self-limiting hepatitis, 21 (75%) were hepatitis B e antigen (HBeAg) positive, 26 (93%) were HBV-DNA positive, and one patient (3.6%) was negative for both markers on admission to the hospital. HBV-DNA cleared after HBeAg clearance in 20 (71.4%), before HBeAg clearance in five (17.9%) and simultaneously with the loss of HBeAg in the remaining two (7.1%) of the 27 initially HBV-DNA- and/or HBeAg-positive patients. Moreover, HBV-DNA remained detectable in serum for 13.3 +/- 6.6 (range: 4-22) days after the appearance of anti-HBe in 71.4% of these patients. In contrast, HBV-DNA and HBeAg remained persistently positive in the two patients who developed chronic HBV infection. These data show that (1) viremia frequently persists after disappearance of HBeAg and (2) appearance of anti-HBe does not indicate the cessation of HBV replication in adults with acute self-limiting hepatitis B.

[Seroepidemiological study on hepatitis C virus infection among blood donors from various regions in China].

A total of 2,273 blood donors from various regions in China were tested for serum anti-HCV antibodies in a seroepidemiological study. The prevalence of anti-HCV in Volunteer blood donors was 0-1.10%, which was lower than that in professional blood donors from Liaoning and Anhui Provinces (1.49% and 3.14%, respectively), whereas the positivity rate of anti-HCV was as high as 30.13% in the professional blood donors from Hebei Province and 31.86% in those from Inner Mongolia Autonomous Region. The prevalence of anti-HCV was significantly higher in the blood donors with history of hepatitis and abnormal ALT levels than those without hepatitis and with normal ALT. Plasma donation was the main cause of HCV infection. However, the prevalence of anti-HCV showed no significant sex and age differences even though the anti-HCV activity profile showed geographic difference.

Prevalence of hepatitis C viral antibody in transfused and nontransfused Egyptian children.

Hepatitis C (HCV) virus is recognized as the major cause of what was previously referred to as parenterally acquired (blood-mediated) non-A, non-B hepatitis. A study involving 252 transfused and nontransfused Egyptian children was conducted from November 1990 through February 1991 to determine the prevalence of HCV and the role of blood and blood and blood product transfusions in the spread of the virus. Serum specimens were assayed by a second generation enzyme immunoassay and were considered reactive only after supplemental testing using the second generation recombinant immunoblot assay. Prevalence among 84 young study subjects with hematologic disorders was 55% (46 of 84), while no HCV antibodies were detected among the two nonhematologic pediatric populations studied: 84 hospital admissions and 84 acutely ill but otherwise healthy outpatients (seeking treatment for symptoms associated with a new condition less than three weeks old in the absence of any chronic health problem). Ninety-two percent (77 of 84) of the hematology-related cases had medical histories of multiple transfusions. Positive antibody responses (46) were significantly associated with increased duration of illness (P < 0.001) and the volume and number of transfusions (P < 0.01) when compared with negative ones (38). However, prior hospitalization and/or surgery were not related to HCV antibody status. The high prevalence of HCV antibody among multiply transfused infants and children suggests that blood and blood product supplies should be regularly screened for HCV antibody.

[Prevalence of HBV and HCV infections in plasma donors with recently increased ALT level].

HCV and HBV markers in sera were examined in a group of donors (71 cases) who had a history of elevated ALT, and were compared with another group of blood donors (217 cases) who had no such a history. The results showed that the positive rates in both two groups of donors were 14.08% and 6.45% for anti-HCV, 3/8 and 2/13 for anti-HCV-IgM in anti-HCV positive cases, 2.82% and 0.46% for HBsAg, 32.39% and 25.35% for anti-HBs, 45.07% and 33.18% for anti-HBc, respectively (P < 0.05-0.01). It showed a good correlation between the positive rates of HCV and HBV markers and the times of ALT elevation.

Hepatitis C virus infection and liver disease in children with thalassemia.

Antibody to the recently identified HCV and type of liver disease were investigated in 25 children with homozygous beta thalassemia. All patients were tested at regular intervals by second generation ORTHO-ELISA test. 19/25 (76%) patients were found repeatedly positive: in 11 cases seroconversion from anti-HCV- to anti-HCV+ status could be also documented. Liver disease was more severe in anti-HCV positive children. Seroconversion however did not produce major changes in liver disease outcome.

Preparation and immunogenicity of an inactivated hepatitis A vaccine.

A hepatitis A vaccine was prepared by formaldehyde inactivation of purified hepatitis A virus (HAV) LSH/S strain grown on human diploid MRC-5 cells. The vaccine was devoid of residual infectivity in vitro and failed to induce in marmoset monkeys any pathological features or variations of haematological and clinical chemistry values. Infectious HAV particles were not detected in faeces and sera of the vaccinated primates by ELISA or after passages in MRC-5 cells. The immunogenicity of the vaccine was evaluated by injecting guinea-pigs with 0.8, 0.2 or 0.05 micrograms of HAV antigen adsorbed onto 0.5 and 1 mg of Al (OH)3 or 0.3 mg of AlPO4. The antibody response, measured by a competitive radioimmunoassay, was dose- and adjuvant-dependent. One injection of 0.2 micrograms of AlPO4-adsorbed HAV antigen induced seroconversion in 100% of animals and high levels of specific and neutralizing serum antibodies. A further increase of antibody titres was observed after the second and third inoculations. These results show that this vaccine formulation is safe and immunogenic in animal models, and suggest that it should be evaluated further by human clinical studies.

Post transfusion hepatitis: comparison between homologous and autologous blood transfusion.

OBJECTIVES: To assess whether the use of autologous blood transfusion may affect the incidence of post-transfusion hepatitis in transfused patients who undergo cardiac surgery. METHODS: One thousand one hundred and twelve polytransfused patients having undergone cardiac surgery were studied from October 1982 through September 1990. Patients were transfused with homologous blood from selected volunteer donors; autologous blood collection or blood saving were introduced in September 1986. Routine laboratory tests were carried out upon hospitalization and monthly for a six-month period. Patients with hepatitis were followed for at least 24 months and liver biopsy was performed in those with chronic hepatitis. RESULTS: Ninety-four (9.8%) of the 959 polytransfused patients developed non-A, non-B post-transfusion hepatitis; anti-hepatitis C virus antibodies were present in 52 out of the 72 patients tested. The mean incubation period for post-transfusion hepatitis was 70 days; hospitalization was required in 47.9% of the patients. The mean number of transfused units was 12.9 in patients who developed post-transfusion hepatitis and 6.96 in the those who did not. Hepatitis was chronic in 42% of the 94 patients; in the others alanine aminotransferase levels normalized in a mean period of 10.3 months. None of the 237 patients who received autologous blood had hepatitis. CONCLUSION: In our study the role of surgical teams in preventing post-transfusion hepatitis was shown to be essential. The high percentage of chronicity and symptom-free hepatitis observed is a further reason to reduce homologous blood transfusions and instore careful follow-up of polytransfused patients.

Reactogenicity and immunogenicity of inactivated hepatitis A vaccines.

Two inactivated hepatitis A virus (HAV) candidate vaccine strain were tested, derived from strains CLF and HM175. Neither vaccine increased liver enzymes levels and reactogenicity was similar to that observed with other alum-absorbed products. Antibody responses were dose-dependent and protection against HAV can be presumed to last for at least three years. All persons receiving 720 ELISA units (El.U) of the CLF vaccine seroconverted after one dose. For the HM175 vaccine, anti-HAV persisted until month 12 after injections at months 0 and 1, suggesting that the third dose of vaccine could be given at any time from month 6 to 12. A double dose of HM175 vaccine (1440 El.U) given as a single bolus resulted in 100% seroconversion by day 14 with a geometric mean anti-HAV level of 121 mIU/ml. This implies that rapid protection can be induced using large doses of inactivated HAV vaccine should time constraints dictate such an approach.