ABSTRACT
Objective To study the roles of apoptosis genes bcl 2, bax, and bak in the pathogenesis of hepatitis D. Methods Expressions of HDAg, bcl 2, bax, and bak in liver specimens of 77 patients with hepatitis D were studied by immunohistochemical method. Meanwhile, the relationship of HDAg expression with the expressions of bcl 2, bax, and bak was studied by double labelling. Results Bcl 2 was mainly expressed in the cytoplasm of hepatocytes, and bax and bak mainly in the cytoplasm of hepatocytes and partly in the nucleus of hepatocytes, and HDAg mainly in the nucleus of hepatocytes. Lots of HDAg and bax/bak positive cells were distributed in infiltrating lymphocytes at the periportal region especially at the advancing edges of areas of piecemeal necrosis. Apoptosis of many hepatocytes was found to locate near the HDAg positive cells. There was positive correlation between the expression of bax/bak and HDAg expression ( P
ABSTRACT
Objective To study the probability of using hepatitis D virus (HDV) ribozyme as a kind of anti-hepatitis-C-virus (HCV) gene thera-py drugs. Methods The natural HDV genomic ribozyme′s stem Ⅳ was optimized and its substrate-binding region reconstructed, thus three recombinant HCV-specific HDV genomic ribozymes RzC1, RzC2 and RzC3 were obtained. HCV RNA 5'-noncoding region and 5'-fragment of C region (HCV RNA5'-NCR-C) were transcribed from plasmid pHCV-neo by T7 phage RNA polymerase in vitro, and radiolabelled at its 5'-end. The trans-cleaving reaction was performed by mixing the ribozymes and substrate at mol ratio 100∶1 under conditions as follows: 37℃, pH7.5, Mg2+ 20 mmol/L and deionized formamide 2.5 mol/L. Percentage of trans-cleaved products were calculated at different time points and used as the activity indicator of the three ribozymes. Results RzC1, RzC2 trans-cleaved more substrate when the time extended, and got to 24.9%,20.3% after reac-ting for 90 minutes respectively; RzC3 was not able to trans-cleave its substrate. Conclusion Recombinant HDV genomic ribozymes have the ability to trans-cleave HCV RNA, but the appropriate target sequence should be selected.
ABSTRACT
Objective To study the role of bcl-2 and bax in the pathogenesis of hepatitis D. Methods Expression of HDAg， bcl-2 and bax in liver specimens of 79 patients with hepatitis D were studied by immunohistochemistry technique. Meanwhile， the relationship between expression of HDAg and that of bcl-2／bax in liver specimens of patients were studied by double labelling technique and serial sections.Results bcl-2 was mainly expressed in the cytoplasm of hepatocytes， bax mainly in the cytoplasm of hepatocytes and partly in the nucleus of hepatocytes， and HDAg mainly in the nucleus of hepatocytes. Most of HDAg and bax positive cells were distributed among infiltrating lymphocytes at the periportal region especially at the advancing edges of areas of “piecemeal necrosis”. Most of hepatocytes of bax positive was found to locate near to positive cells of HDAg， and there were positive correlations between degrees of bax expression and HDAg expression（P＜0.05）. Conclusions The distribution and the expression of bax and HDAg are significantly correlated with the activity of inflammation and the severity of the liver damage， and HDV infection may induce the expression of bax in the hepatocytes，and hepatocyte apoptosis mediated by bcl-2／bax may play an important role in the liver cell injury by HDV infection.
ABSTRACT
Objective To investigate the possible relationship between the vitamin E(VE) contents in the plasma and development of hepatosis.Methods VE and malondialdehyde(MDA) contents in 217 cases infected by various types of viral hepatitis were measured by means of fluorescence and TBA.Moreover,the relationship between both of the contents and the alteration of liver function was observed dynamically.Results The plasma levels of VE in all cases were decreased obviously.Interestingly,lower the contents of VE were,more severity the disease was.Howevey,the levels of VE recovered step by step when the liver function return to normal in the acute hepatitis cases.Unfortunately,the levels in the chronic hepatitis cases were higher in the recovering phase than in the acute phase,but still lower than normal.Moreover,VE kept low levels and showed a negative relationship to the bilirublin in both worsening and dying cases.Conclusion To observe the VE contents is helpful to the clinical evaluation of the hepatosis progress and prognosis.
ABSTRACT
Objective To investigate the influence of vitamin E on type Ⅰ collagen, messenger RNA expression of transforming growth factor ? 1 (TGF? 1) and tissue inhibitor of matrix metalloproteinases-2 (TIMP 2) in hepatic stellate cells (HSC). Methods HSC s were cultured in vitro, the effect of vitamin E on type Ⅰ collagen was observed by mean of immunohistochemistry; mRNA expression of TGF? 1 and TIMP 2 were investigated with in situ hybridization. Results In experiment group, vitamin E markedly inhibited expression of type Ⅰ collagen of HSC as compared with control group, but could not suppress mRNA expression of TGF? 1 and TIMP 2.Conclusions Anti-fibrotic effect of Vitamin E may be implemented through inhibiting the expression of type Ⅰ collagen.
ABSTRACT
A 27-peptide,a fragment of hepatitis delta antigen(HDAg),was synthesized and used to develop an ELISA method for ihe detection of anti-HD.It was found that positive anti-HD reaction occurred between the coated 27-peptide and a stored sample of serum which was known anti-HD positive.Absorption test revealed that the synthetic peptide competed with natural HDAg for anti-HD,suggesting that the peptide possessed the antigenicity similar to that of natural HDAg.The antigenicity of the synthetic peptide was quite specific wihtout cross reaction with normal human and mouse sera and with anti-HA.anti-HB and anti-HC sera.Among 300 blood donors,there was only 1 case(0.33%)anti-HD positive with an ALT level 2 times higher than normal.In 41 cases of non-B hepatitis and 52 cases of HAV hepatitis,none was anti-HD positive.In 211 cases of various types of HBV hepatitis,21 were(9.95%)anti-HD positive,among whom 2/82(2.5%)werehealthy HBV carriers,6/43(13.95%)were patients with a-cute icteric hepatitis,6/60(10.00%)were patients of chronic active hepatitis,4/18(22.20%)were patients of severe hepatitis,and 3/8(37.50%)were those with liver cirrhosis.These results were consistent with those in our previous reports.
ABSTRACT
The effects of human fetal hepatic stimulator substance (h-HSS) and liver cytosol (h-FLC) on the survival rate,the intensity of liver damage,and liver regeneration in rats with liver necrosis induced with D-galactosamine were observed.The physico-chemical properties of h-HSS were preliminarily determined.It was found that both h-HSS and H-FLC could markedly increase the.survival rate of the rats with toxic liver necrosis,significantly decrease the serum level of ornithine carbamoyl transferase and mitochondria aspartic trans-aminase,improve the hepaplastin activity,and elevate the incorporation rate of 3H-thymidine with liver DNA.h-HSS was stable after heated to 95℃ for 30 minutes,and sensitive to trypsin treatment.It demonstrated one major band at 15 000 daltons and 3 minor ones at 24 000,34 000,and 40 000 daltons respectively on sodium dodecyl sulfate-polyacrylamide gel electrophoresis.The relative content of the major band was 47.57%.
ABSTRACT
The in situ hybridization of HBV DNA with biotinylated probe in formalin-fixed and paraffin-embedded liver tissue sections was performed and the localization and morphology of HBV DNA in various types of chronic liver diseases was observed. 25 cases out of 75 (asymptomatic HBsAg carriers 4/21, chronic persistent hepatitis 2/7, chronic active hepatitis 10/21, liver cirrhosis 0/3, subacute severe hepatitis 4/11 and hepatocellular carcinoma with cirrhosis 5/12) were HBV DNA positive(33.33%). HBV DNA in hepatocytes was demonstrated as numerous coarse grains in cytoplasm and occasionally a few grains in individual nucleus. The distribution of the HBV DNA positive hepatocytes in the sections was in 3 forms, i.e., the diffuse type(Type I, 4/25, 16%), the focal type(Type II, 20/25, 80%), aud the scattered type (Type III, 1/25, 4%). The HBV DNA grained in the cytoplasm might be condensed (Type a)or sparsed(Type b)depsnding on the number of HBV DNA grains. The different distribution patterns and the amount of HBV DNA might be a reflection of the replication activity and the state of infectivity of HBV. HBV DNA positive hepatocytes were found only in the tissue adjacent to the cancer cells in cases of hepatocellular carcinoma with cirrhosis.
ABSTRACT
The paper is to report a retrospective study on delta antigen infection in 35 patients with severe hepatitis B (13 subacute and 22 chronic cases).One serum sample from each patients was collected at least 21 days (21～156 days)after the last attack of the disease,and anti-delta antigen was tested with EIA method (Abbot kit made in USA).The upper limit of O.D.value for positive resultswas calculated as 0.391.It was found that anti-delta antigen was positive in 12 cases(34.3%)out of the 35. 23.1%(3/13)of the subacute and 40.9%(9/22) of the chronic cases were positive.There were 3～5 serological parameters indicating HBV infection in each case.All the 12 positive patients were males of Han Nationality with a mean age of 34 years (a range of 19 to 51).11 were from Sichuan province and only 1 from Guizhou.3 were subacute cases without previous history of hepatitis,and aggravation of the clinical condition occurred in their course;there was a history of acute icteric hepatitis in the remaining 9 cases and 5 out of the 9 were long-term HBsAg carriers.The recent attack of the disease often showed a rapid progression to the severe stage within a month.It is suggested that delta antigen might be one of the factors leading to the aggravation of viral hepatitis.
ABSTRACT
The immunohistocheinical observation of & antigen in liver sections from 27 cases of severe hepatitis B (SHE) was performed with ABC method and the result was compared with that of & antibody in the sera. 6 cases of SHB were & antigen positive in liver sections (6/27, 22.22%) . It showed a high attack rate of ? agent in SHE cases in Chongqing area. The detective rate for 5 agent infection could be increased by the immunohistochemical measurement. There was positive staining in both nuclei and cytoplasm in all the 6 cases.
ABSTRACT
Anti-HAV-IgM, anti-HBc-IgM, and three antibody systems of hepatitis B were tested in 48 patients with fulminant hepatitis to determine which of the hepatitis viruses A and B was the causative agent. HBV was found in 31 cases, HAV in one case, mixed infection of HBV and HAV in 15 cases (31.25%),and uncertain agent in 1 case.Ten cases out of the 15 with mixed infection were subacute, and the other 5 chronic. 13 patients were male and 2 female. The age of the patients ranged from 15 to 49 years with an average of 30.39. The clinical conditions of 10 cases became more serious and some of them even died during hospitalization. The clinical and biochemical parameters of the patients with mixed infection showed no statistical difference from those of the patients with single infection of either HBV or HAV of the same time period. 7 patients complained of sudden aggravation of their clinical conditions after they exhibited a group of symptoms and signs of "acute icteric hepatitis". One such case was found during his hospitalization and his anti-HAV-IgM turned positive at the same time. This patient died eventually.It is considered that aggravation of acute or chronic hepatitis can be resulted from superinfection and this problem deserves close attention.
ABSTRACT
A study on the optimal dilution of serum to detect anti HBc-IgM with ELISA. was carried out. Samples were diluted to 10-1to 10-7. Among 26 samples of normal individuals, 22 were negative, but two samples of 10-1 dilution, one sample of 10-2, and one sample of 10-1-10-3 were positive though the control wells were all negative. 42 samples of the patients with variours types of hepatitis B were examined. It was found that when serum dilution was 10-1, only 6 cases were positive; when the dilution was 10-2, 20 cases were positive; when the dilutions were 10-3, 5?10-3, and 10-4, high positive rates were obtained and they were 88.1%, 78.5%, and 69% respectively; and when the samples were further diluted, the positive rate decreased.In order to get goodfresults and minimize false results, either false negative or false positive, it is suggested that 5?10-3 be the optimal dilution.
ABSTRACT
This paper is to report the result of using ELISA to detect HBsAg-IgM complex in the serum of patients with viral hepatitis B. A series of serum samples of 35 patients,including 29 acute,2 subacute severe and 4 chronic active cases were tested. 28 normal individuals were also tested as control.HBsAg-IgM complex was discovered in the sera of all the patients in the first one or two weeks of hospitalization. It disappeared from the sera of 24 patients (among whom 10 have been followed up for more than 3 to 6 months) who recovered completely after 4 weeks of treatment. The disappearance of the complex occurred long before the clearance of HBsAg and the returning to normal levels of other biochemical parameters. In 5 cases of acute hepatitis with the tendency to be chronic and in those subacute and chronic cases,HBsAg-IgM complex delayed to disappear and could be found in the sera even 6 months later. All the normal controls were negative.It is suggested that the detection of HBsAg-IgM complex in the serum of hepatitis B patient may be used not only as a criterion to evaluate the prognosis of acute cases but also as an index to determine whether the patient is suffering an active disease process.
ABSTRACT
The effects of different vitamin E(VE) containing diets on lipid peroxi-dation and the degree of liver damage in rats were observed in the model of liver necrosis induced by galactosamine(D-GAL).The results were as follows: After 8 weeks, the basic levels of VE in plasma and liver tissue of the group fed VE-supplemented diet were significantly higher than that of the group fed normal VE diet, in the company with a significant lower basic plasma MDA level. At each time interval after D-GAL injection, the plasma and liver VE levels were found higher in the VE-supplemented group, and the content of GSH as well as the activities of SOD, GSH-Px, HPT were maintained satisfactorily while the MDA content and OCT, m-AST activities were very significantly lowered and the pathological changes of the liver were not so severe as compared with that in the normal VE diet group. It was noteworthy that the changes of almost all the parameters metioned above in the group fed VE-deficient diet went to an opposite direction to the VE-supplemented group. The results indicated that the VE-supplemented diet has a good liver protective effect, which may be established on the increased VE content and suppressed lipid peroxidation in the liver. VE-deficient diet could sharply reduce the VE content in the body and significantly intensify the susceptibility of liver to the harmful agents.