[Lamivudine and entecavir significantly improved the prognosis of early-to-mid stage hepatitis B related acute on chronic liver failure].

OBJECTIVE: To clinically study the antiviral effects of lamivudine and entecavir on patients with early-to-mid stage Hepatitis B related acute on chronic liver failure (HBV-ACLF). METHODS; A prospective, randomized, open and parallel controlled clinical trial was designed to observe the antiviral effects of nucleoside analogues on patients with early-to-mid stage HBV-ACLF. Three groups were set for controlled study, i. e. basic treatment group, lamivudine plus basic treatment group and entecavir plus basic treatment group. RESULTS: One month after treatment, the improvement rates of lamivudine group and entecavir group were 58.85% and 59.15% respectively, significantly higher than that of basic treatment group which was 34.84% (Chi(2) = 9.8323, P = 0.043). By the end of six months, the cumulative survival rates of patients with the antiviral treatments, i.e., lamivudine, entecavir, were 65.8%, 60.1%, significantly higher than that (42%) without the antiviral treatment (P = 0.045, P = 0.04 respectively). The cumulative survival rate in patients with a MELD score < 30 was higher than that with a MELD score over 30 (Chi(2) = 3.920, P = 0.048). For the patients with pretreatment HBV DNA > or = 10(7), the cumulative survival rate in patients with entecavir treatments group was higher than that of patients in basic treatment group (Chi(2) = 5. 014 P= 0.025). According to the Ordinal Regression analysis, antiviral therapy by using either lamivudine or entecavia could significantly increase the improvement rate of patients with early-to-mid stage HBV-ACLF. But severe complications, including hepatorenal syndrome, electrolyte imbalance and hepatic encephalopathy, medical history of liver cirrhosis, and pretreatment HBV DNA > or = 10(7) had significant impacts on prognosis of this group patients. CONCLUSIONS: Antiviral therapy by using either lamivudine or entecavia could significantly increase the survival rate of patients with early-to-mid stage HBV-ACLF.

[Relation of viral genotypes to clinical features in children with chronic hepatitis B].

OBJECTIVE: To investigate the relation of HBV genotypes to clincal features in children with chronic hepatitis B. METHODS: The genotypes of serum HBV DNA from 404 children with chronic hepatitis B were determined by PCR using type-specific primers. RESULTS: For the 404 children, genotype B in 99 (24.5%), genotype C in 285 (70.5%). For the 75 children from south part of China, 29 were of genotype B (38.7%) and 44 of genotype C (58.7%). For the 329 children from north part of China, 70 were of genotype B (21.3%) and 241 of genotype C (73.3%). There were significant differences between the children from south part and those from north part of China in genotype B and C (P = 0.002). Genotype B and C were not significantly correlated to gender, age and mother-to-fetus transmission. There was no marked difference in liver injury severity (P = 0.4796), serum HBeAg positivity, HBVDNA level, inflammatory degree of liver tissue (P = 0.209) and liver fibrosis( P = 0.177) between the children with genotype B and those with genotype C. CONCLUSION: In children with HBV infection, genotype C accounts for 70.5% and genotype B for 24.5%. The genotypes are of regional difference in children with HBV infeciton. There are replication and liver pathological change between genotype B and genotype C.

[Etiological analysis of 1977 patients with acute liver failure, subacute liver failure and acute-on-chronic liver failure].

OBJECTIVE: To investigate the etiology of 1977 patients from northern China with acute (ALF), sub-acute (SALF) or acute-on-chronic liver (ACLF) failures. METHOD: The age, gender, etiology, pathogenesis, and prognosis of the 1977 patients with liver failures were retrospectively analyzed. RESULTS: Of the 1977 cases, the three most common causes of ALF were HEV (33.96%) or HBV (13.21%) infections or those caused by medicines (9.43%). The three predominant causes of SALF were medicines (31.53%), HEV (16.22%) or HBV (9.91%) infections, but those of the ACLF were HBV (90.29%) infection, alcoholic hepatopathy (2.65%), and HBV super infected with HEV (2.26%) infections. 90.09% (1781) patients were infected by hepatotropic viruses. Of these 1781 patients, the most common cause of their liver failures was HBV infection (92.93%). In these HBV infected patients, 77.10% were from 26 to 55 years old. From 2005 to 2007, there were 39 patients with alcoholic liver failure. In the past two years, there were 23 patients with drug induced liver failure. The improvement rate of the 1977 patients after their treatments was 35.56%. The improvement rate of HEV infected liver failure was higher than drug induced liver failure (P less than 0.05); no statistical significance was found between other groups (P more than 0.05). CONCLUSION: Different types of liver failure have different predominant causes. HBV infection is the most common cause in our 1977 patients. In the past two years, the number of drug induced liver failures and alcoholic liver failures have been increasing.

[Efficacy and safety of Fufang Biejia Ruangan tablet in patients with chronic hepatitis B complicated with hepatic fibrosis].

OBJECTIVE: To study the clinical therapeutic effects and safety of Fufang Biejia Ruangan tablet (FBRt) in patients with chronic hepatitis B complicated with hepatic fibrosis. METHODS: Totally 420 patients were randomly divided into two groups, FBRt group (300 cases) were treated with Fufang Biejia Ruangan tablets and control group (120 cases) were treated with He Luo Shu Gan capsule, the patients in both groups were treated for 6 months. RESULTS: The cure rate and total effective rate of FBRt group were significantly higher than those of control group (55.67 percent and 81.67 percent vs. 15.8 percent and 60.00 percent, P less than 0.01). CONCLUSION: Fufang Biejia Ruangan tablet could alleviate clinical symptoms and hepatic fibrosis. Fufang Biejia Ruangan tablet is effective and safe in treatment of patients with chronic hepatitis B complicated with liver fibrosis.

[To re-evaluate the clinical classification criteria of liver failure].

OBJECTIVE: To study the clinical feature and more reasonable diagnostic typing criteria for patients with liver failure. METHODS: 13/21 cases of ALF, SALF with no past liver disease, 49/72 cases of with chronic hepatitis, and 23/73 cases ALF, SALF with liver cirrhosis, were analyzed respectively. RESULTS: 1 ALF patients (1). There exist significant statistic differences in ALB, ALT, CHE in three ALF groups.(2). It had statistic differences in those patients with hepatic encephalopathy.(3). The prognosis of the patients with chronic hepatitis group (42.85 percent) was best than that of chronic cirrhosis (26.09 percent) and no past liver disease (15.38 percent). (2) In SALF patients (1). There exist significant statistic differences in ALB, GLO, ALT, AST, BDIL, GLU and CHE in three SALF groups.(2). It had statistic differences in those patients with hepatic encephalopathy in three SALF groups.(3). The prognosis of the patients with chronic hepatitis group (51.39 percent) was best than that of chronic cirrhosis (36.85 percent) and no past liver disease (33.33 percent). CONCLUSION: There are different clinic feature and prognosis in three ALF or SALF groups, so we suggest that it were clinic practicability and science in classify of liver failure at present.

[Reevaluation of the typing criteria for patients with chronic severe hepatitis].

BACKGROUND: To study the clinical features and more reasonable typing criteria for patients with chronic severe hepatitis and decompensated liver function. METHODS: Data of 106 cases of decompensated cirrhosis, 124 cases of chronic liver failure and 100 cases of chronic liver failure (chronic liver failure group I, CLF I) with decompensated cirrhosis (chronic liver failure group II, CLF II) were analyzed retrospectively. RESULTS: (1) The ages were youngest in chronic liver failure group I (about 30 years), and the oldest in decompensated cirrhosis group (about 50 years). (2) There were significant differences in albumin, globulin, ALT, AST, protruding activity, blood glucose, blood lipid and cholinesterase among the three groups. (3) There was no significant difference in upper digestive tract bleeding and hepatorenal syndrome, on the other hand, there was significant difference in ascites and hepatic encephalopathy. (4) The prognosis of the patients in decompensated cirrhosis group was better than that of chronic liver failure group I and chronic liver failure group II. CONCLUSION: The clinical feature and prognosis in three groups were different, so, it is suggested that chronic severe liver disease be divided into 2 types: one is chronic severe liver disease type I, which is associated with chronic hepatitis, and the other is chronic severe liver disease type II, which is associated with cirrhosis, and the typing criteria for decompensated cirrhosis remains unchanged.

[Analysis on prognostic factors of liver failure in children].

OBJECTIVE: To analyze the prognostic factors of liver failure in children. METHODS: The clinical data of 105 children with liver failure treated in the No. 302 Hospital in the past 17 years were retrospectively analyzed. The related factors were analysed by EXCELL 2000 and STATA 7.0, multivariate statistical analysis was performed by Logistic regression. RESULTS: (1) A total of 72 children died and the mortality was 68.6%. (2) Univariate statistical analysis showed that the factors significantly correlated with death were age, clinical type and stage of liver failure, decrease in prothrombin activity (PTA) and albumin (AIB) level, increase in serum level of total bilirubin (TBIL), appearance of deviation of TBIL and ALT, complications and hepatic encephalopathy. There was no significant difference between boys and girls. (3) There was no significant difference among etiological diagnoses such as HBV infection, Wilson's disease, and unknown pathogeny. (4) Multivariate statistical analysis showed that PTA (P = 0.000) and TBIL (P = 0.029) were independent risk factors of mortality of the children. CONCLUSION: The prognosis of liver failure in children is poor and mortality is high. PTA and TBIL might be useful for indicating prompt diagnosis and treatment to improve survival rate of the children with liver failure.

Pathological changes and clinical manifestations of 1020 children with liver diseases confirmed by biopsy.

BACKGROUND: Liver biopsy plays an important role in accurate diagnosis of various liver diseases in children and liver damages caused by systemic illnesses. This study was designed to evaluate the value of liver biopsy in diagnosis of liver diseases in children and explore the relationship between their pathological changes and clinical manifestations. METHODS: One-second liver biopsy was performed in 1023 pediatric patients with liver diseases at our department from 1983 to 2000. Diagnosis of viral hepatitis was based on the diagnostic criteria formulated by the Chinese Society of Infectious and Parasitic Diseases in 1995. Inflammatory changes of the liver were graded from 0 to 4 (G0-4). RESULTS: Liver biopsy was performed successfully in 1020 patients including 135 infants and young children, of whom 90% were hospitalized patients with chronic liver diseases. Hepatitis virus was the leading cause for chronic liver diseases, among which hepatitis B was detected in 75.4% of the patients. Sixty-nine patients showed liver impairment induced by disorders relevant to that metabolism, Wilson's disease, and glycogen storage disease. Liver inflammatory injury (

[Characterization of peripheral dendritic cell subsets and its implication in patients infected with severe acute respiratory syndrome].

OBJECTIVE: To investigate the dynamic changes of dendritic cell subsets in peripheral blood of patients infected with severe acute respiratory syndrome (SARS) and evaluate their roles in the immunopathogenesis of SARS. METHODS: Flow cytometry was applied to study the dynamic alteration of the number and frequencies in circulating DC cell subsets in 30 SARS patients including critical SARS (n = 11) and general SARS (n = 19). The reasons and clinic significances of the peripheral blood DC subsets changes in SARS patients were also analyzed in our study. RESULTS: The patients in critical status had a 9-week course of disease, longer than the 6-week course observed in subjects in general status. The frequency of peripheral DC cell subsets significantly dropped beginning from the onset of symptom in SARS patients and was maintained at significant low levels during the following 4 - 5 weeks, 1.7 +/- 1.8, 5.3 +/- 5.0/ micro l for DC1, 0.57 +/- 1.02, 0.98 +/- 1.11/ micro l for DC2 for cases in critical and general statuses, respectively, compared with healthy subjects; more importantly, the pDC2 even disappeared in the patients who died from SARS diseases. The possible reasons responsible for the alteration of DC subsets in peripheral blood is likely to be the direct attack of SARS-CoVin circulation and be partially involved the application of large dose of steroid. The frequency in DC cell subsets returned to normal level in convalescent stage. CONCLUSION: Our results showed SARS patients had a significant decrease of circulating DC cell subset frequency, which maybe lead to the host immunodeficiency response to SARS-associated coronavirus (SARS-CoV).

[Clinical features of liver failure in children].

OBJECTIVE: To analyze the etiology, clinical and laboratory characteristics of hepatic failure in 105 children. METHODS: The clinical data of 105 children with hepatic failure treated in our hospital from January 1986 to June 2003 were retrospectively analyzed by EXCELL 2000 and t test. RESULTS: (1)Of the 105 children with hepatic failure, 9 were cases with fulminant hepatic failure, 38 with subacute hepatic failure and 58 with chronic hepatic failure. (2)Morbidity was the highest in 7-12 years old children (43/105, 41.0%) followed by infants (30/105, 28.6%). (3)CMV infection could be confirmed in 9 infants (30.0%), etiological diagnosis was not possible in 13 infants (43.3%). Etiological diagnosis could be confirmed in children over 1 year of age, which included hepatitis B (n=22, 29.3%), Wilson's disease (n=15, 20.0%), hepatitis A (n=10,13.3%). Etiology in 21 cases (28.0%) could not be confirmed. (4)Seventy-one cases (67.6%) had ascites, 34 of them (47.9%) had spontaneous peritonitis. Thirty-five cases were complicated with other infections. The commonest complication was pulmonary infection and sepsis was the next. Fifty-one cases (48.6%) had hydroelectrolyte imbalance. Forty-eight cases (46.2%) had hepatic encephalopathy, which may be subclinical in children under three years of age. (5)The incidence of hypoglycemia was 77.2%(71/92). CONCLUSION: The etiology of liver failure was related to age. CMV infection was the commonest in infants. HBV, HAV infection was the commonest in children over 1 year of age and Wilson?s disease was the next. It is necessary to prevent and manage the associated complications as early as possible such as spontaneous peritonitis, hepatic encephalopathy, hydroelectrolyte imbalance and hypoglycemia etc.

Screening of augmenter of liver regeneration-binding proteins by yeast-two hybrid technique.

OBJECTIVE: To investigate the biological function of augmenter of liver regeneration (ALR), we used yeast-two hybrid technique to detect proteins in hepatocytes interacting with ALR. METHODS: ALR bait plasmid was constructed by using yeast-two hybrid system 3, then transformed into yeast AH109. The transformed yeast was mated with yeast Y187 containing liver cDNA library plasmid in a 2XYPDA medium. Diploid yeast was plated on a synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selection and screening. After extracting and sequencing of the plasmid from blue colonies. Analysis was performed by bioinformatics. RESULTS: Of 36 colonies sequenced, 14 are metallothionein, 12 albumin, and 3 selenoprotein P. One colony is a new gene with unknown function. CONCLUSION: The successful cloning of gene of ALR interacting protein has paved the way for studying the physiological function of ALR and associated proteins.

[Cloning of genes transactivated by hepatitis B virus X protein].

OBJECTIVE: To construct a subtractive cDNA library of genes transactivated by hepatitis B virus X protein (HBX) using suppression subtractive hybridization (SSH) technique and to clone genes associated with HBX transactivating function. METHODS: The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-X and pcDNA3.1(-) empty vector respectively, then cDNA was synthesized. After restriction enzyme RsaI digestion, a number of small size cDNA was obtained. Then tester cDNA was subdivided into two portions and each was ligated with different cDNA adaptor. After tester cDNA was hybridized with driver cDNA twice and underwent nested polymerase chain reaction (PCR) twice the production was subcloned into T/A plasmid vectors to set up the subtractive cDNA library. Amplification of the library was carried out with E. coli strain JM109, some cDNA was sequenced and analyzed in GenBank with Blast. RESULTS: The subtractive cDNA library of genes transactivated by HBX was constructed. The amplified library contained 85 positive clones, and colony PCR showed that these clones contained 200-1000 bp inserts. 65 clones were analyzed by sequencing and bioinformatics, which suggested nineteen known genes and fifteen genes with unknown function. CONCLUSION: A subtractive cDNA library of genes transactivated by HBX using SSH technique has been constructed successfully, which may bring some new clues for studying the biological functions of HBX and the pathogenesis of hepatoma.

Interaction between hepatitis C virus core protein and translin protein--a possible molecular mechanism for hepatocellular carcinoma and lymphoma caused by hepatitis C virus.

AIM: To investigate the interaction between hepatitis C virus core protein and translin protein and its role in the pathogenensis of hepatocellular carcinoma and lymphoma. METHODS: With the components of the yeast two hybrid system 3, "bait" plasmids of HCV core the gene was constructed. After proving that hepatitis C virus core protein could be firmly expressed in AH109 yeast strains, yeast two- hybrid screening was performed by mating AH109 with Y187 that transformed with liver cDNA library plasmids-pACT2 and then plated on quadruple dropout (QDO) medium and then assayed for alpha-gal activity. Sequencing analysis of the genes of library plasmids in yeast colonies that could grow on QDO with alpha-gal activity was performed. The interaction between HCV core protein and the protein we obtained from positive colony was further confirmed by repeating yeast two - hybrid analysis and coimmunoprecipitation in vitro. RESULTS: A gene from a positive colony was the gene of translin, a recombination hotspot binding protein. The interaction between HCV core protein and translin protein could be proved not only in yeast, but also in vitro. CONCLUSION: The core protein of HCV can interact with translin protein. This can partly explain the molecular mechanism for hepatocellular carcinoma and lymphoma caused by HCV.

[Cut-off period of subclassification and pathological features of severe hepatitis based on clinical and pathological analyses].

BACKGROUND: To explore the cut-off period of subclassification and pathological features of severe hepatitis (SH). METHODS: Based on combined clinical and pathological analyses, the complete clinical and biopsy or autopsy liver tissues data from 196 cases of patients with severe hepatitis were investigated. Meanwhile, proliferative hepatocytes, cholangioepithelia and collagens were identified by a panel of monoclonal antibodies such as those against albumin, cytokeratin 18,19 and collagen I, III with immunohistochemical method. RESULTS: The clinical and pathological analyses indicated the cut-off periods of acute, subacute and chronic SH (ASH,SSH and CSH) were (13.4+/-7.2) d, (77.4+/-69.3) d and (80.5+/-63.2) d, respectively. Among all SH cases, one case of ASH patient presented clinical manifestation and pathological changes of ASH for 21 days, however, one patient with SSH was demonstrated 12 day course by histological examination. The time of cut-off period between ASH and SSH in child cases was shorter than that in adult cases. Histologically, ASH liver tissues showed massive and/or submassive necrosis caused by one attack, with congestive sinusoid frameworks and proliferative cholangioepithelium-like hepatocytes, while SSH liver tissues presented combined fresh and old submassive or massive necrosis caused by multiple attacks, accompanied by obviously proliferative bile ducts and sinusoid framework collapse.However, the pathological changes of CSH showed ASH- or SSH-like lesions on the background of chronic liver injury. CONCLUSION: Our data indicated that the cut-off period between ASH and SSH is in accordance with the Scheme of Viral Hepatitis Prevention and Therapy, China, published in 2000, but excluded a part of child SH cases. In our study, the authors found a few pathological features in ASH and SSH.

[Detection of coagulation factor V in patients with severe hepatitis and its clinical significance].

BACKGROUND: To investigate the prognostic significance and role of coagulation factor V (CFV) levels in clinical diagnostic criteria for severe hepatitis. METHODS: The CFV level and prothrombin activity (PTA) were tested by turbidimetry for 129 times in 58 patients with severe hepatitis. Comparative studies and clinical significance of CFV and PTA were analyzed by SPSS and SDAS softwares. RESULTS: 1. The levels of CFV and PTA were 15.3%+/-9.7% and 23.5%+/-10.0%, respectively, at the onset of severe hepatitis. 2. The mortality of severe hepatitis gradually increased with the gradual decrease of CFV or PTA during the most severe stage of the illness (P=0.000). 3. The levels of CFV and PTA decreased continually and rapidly in patients who died but gradually increased in survivors. The decrease or increase of PTA preceded that of CFV on the exacerbation or convalescent stage. 4. Hepatic encephalopathy occurred in 14 cases (24.14%). In 10 cases, it occurred in the terminal stage of the illness, far later than the time of the decrease of CFV. 5. The level of CFV was closely related to PTA (the correlation coefficient was 0.812), the level of CFV was almost consistent with that of PTA. CONCLUSION: 1. The level of CFV is an important prognostic indicator in severe hepatitis and is more specific than PTA. 2. Simultaneous determination of CFV and PTA may be helpful in earlier and more accurate diagnosis of severe hepatitis. 3. Possible use of CFV as one of the criteria for liver transplantation in patients with severe hepatitis should be studied.

Preparation of human single chain Fv antibody against hepatitis C virus E2 protein and its identification in immunohistochemistry.

AIM: To screen human single chain Fv antibody (scFv) against hepatitis C virus E2 antigen and identify its application in immunohistochemistry. METHODS: The phage antibody library was panned by HCV E2 antigen, which was coated in microtiter plate. After five rounds of biopanning,56 phage clones were identified specific to HCV E2 antigen. The selected scFv clones were digested by SfiI/NotI and DNA was sequenced. Then it was subcloned into the vector pCANTAB5E for expression as E-tagged soluble scFv. The liver tissue sections from normal person and patients with chronic hepatitis B and chronic hepatitis C were immunostained with HCV E2 scFv antibody. RESULTS: The data of scFv-E2 DNA digestion and DNA sequencing showed that the scFv gene is composed of 750 bp. ELISA and immunohistochemistry demonstrated that the human single chain Fv antibody against hepatitis C E2 antigen has a specific binding character with hepatitis virus E2 antigen and paraffin-embedded tissue, but did not react with liver tissues from healthy persons or patients with chronic hepatitis B. CONCLUSION: We have successfully screened and identified HCV E2 scFv and the scFv could be used in the immunostaining of liver tissue sections from patients with chronic hepatitis C.

[Epitope mapping of hepatitis C virus non-structure protein 5 from a 7 peptide phage library by using immobilized specific monoclonal antibody].

OBJECTIVE: To screen HCV NS5 mimotopes by using monoclonal antibody and phage peptide library. METHODS: By using HCV NS5 monoclonal antibody as selective molecule, a 7 peptide phage library was biopanned and positive clones were selected by ELISA, competition assay and DNA sequencing. RESULTS: Twelve positive clones were chosen for DNA sequencing. From the experiment and sequencing comparison results, one epitope was confirmed as the mimotope of HCV NS5. CONCLUSIONS: HCV mimotope is obtained by phage peptide library screening. The result provides a new approach for HCV therapy and vaccine development.