Study on the Anti-apoptotic Effect of Curcumin on H 2O2-induced H 9c2 Cardiomyocyte Injury and the Regulation of NF-κB Signaling Pathway / 中国药房

OBJECTIVE：To investigate the anti-apoptotic effect of curcumin on H 2O2-induced H 9c2 cardiomyocyte injury and the regulation of NF-κB signaling pathway. METHODS：H9c2 cardiomyocyte were randomly divided into normal control group ， injury model group ，curcumin low-dose ，medium-dose and high-dose groups （25，50，100 μmol/L）. Normal control group didn ’t received any intervention. The cells in injury model group were induced with 50 μmol/L H2O2 for 12 h to establish the injury model. The cells in curcumin groups were treated with relevant concentration of drugs for 24 h，and then induced with 50 μmol/L H2O2 for 12 h. After cultured for 24 h，survival rate and apoptotic rate of cells were measured by MTT method and TUNEL method ；SOD activity and MDA content were determined by WTS- 8 assay and color test ；relative fluorescence intensity of LC 3 positive expression was detected by immunofluorescence method ；mRNA expression of NF-κB p65 in cells was detected by real-time PCR ； Western blotting assay was used to detect the protein expression of NF-κB p65 and p-NF-κB p65 in cells. RESULTS ：Compared with normal control group ，survival rate and SOD activity were decreased significantly in injury model group ，while apoptotic rate ， MDA content ，relative fluorescence intensity of LC 3 positive expression ，mRNA expression of NF-κB，protein expression of NF-κ B p 65 and p-NF-κB p65 as well as p-NF-κB/NF-κB were increased significantly（P＜0.05）. Compared with injury model group ， survival rates and SOD activities were increased significantly in curcumin groups ，while apoptotic rates ，MDA contents ，relative fluorescence intensities of LC 3 positive expression ，mRNA expression of LC 3 positive cells ，protein expression of NF-κB p65 and p-NF-κ B p65 as well as p-NF-κ B p65/NF-κ B p65 were decreased significantly （P＜0.05）. CONCLUSIONS ：Curcumin can increase the survival rate of H 2O2-induced H 9c2 cardiomyocyte injury ，decrease its apoptotic rate ，increase SOD activity and decrease MDA content in cardiomyocytes. Above effects may be related to the regulation of NF-κB signaling pathway.

Design and activity assessment of HDV ribozymes to trans-cleave HCV RNA / 第三军医大学学报

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.

Study on ICAM-1 expression of primary cultured human hepatocyte regulated by cytokines and dexamethasone / 中华传染病杂志

Objective To study the effects of IFN ?,TNF ?,IL 1 and dexamethasone on the expression of intercellular adhesion molecule 1(ICAM 1) of primary cultured human hepatocyte. Methods ICAM 1 expression of primary cultured human hepatocyte was induced in vitro by IFN ?,TNF ? and IL 1, measuring by cellular enzyme linked immunosorbent assay(ELISA). The effect of dexamethasone on human hepatocyte ICAM 1 expression was observed by adding dexamethasone into the cultured human hepatocytes prior to cytokine stimulation. Results The levels of ICAM 1 expression were significantly lower in unstimulated human hepatocytes than in human hepatocytes induced by IFN ?,TNF ? and IL 1. There were relations among the levels of ICAM 1 expression and cytokine concentration as well as induced period of time. Dexamethasone could partly inhibit the ICAM 1 expression on human hepatocytes induced by TNF ?,IL 1 and IFN ?.The inhibition rates were 40.3%?5.9%,38.1%?4.8%, and 37.6%?6.7%, respectively.Conclusions Enhanced ICAM 1 expression of primary cultured human hepatocyte may be induced in vitro by IFN ?,TNF ? and IL 1, which may be partly inhibited by dexamethasone.

The assay of apoptosis of PBMC and lymphocyte subsets in chronic hepatitis B / 中国免疫学杂志

Objective:In order to explore the AICD happening in PBMC and the situation of peripheral blood lymphocyte subsets in chronic/ chronic severe hepatitis B Methods:The peripheral blood mononuclear cells of patients with chronic /chronic severe hepatitis B were cultured with PHA P for 72 h Then the apoptosis of PBMC was assayed by flow cytometry The peripheral blood lymphocyte subsets of patients with chronic /chronic severe hepatitis B were assayed by flow cytometry and automatic blood analyzer Results:The percentage of apoptotic PBMC in chronic hepatitis B group was higher than that in chronic severe hepatitis B group(P

Investigation of HBV-DNA in peripheral blood mononuclear cells and hepatocytes with molecular hybridization / 第三军医大学学报

The existence of HBV-DNA in the mononuclear cells of peripheral blood and in the serum of 61 patients with HBV infection was determined with southern blot and dot blot hybridization,and that in the liver tissue of 31 patients out of the 61 with southern blot and in situ hybridization.The positive rate of HBV-DNA in the serum,mononuclear cells and hepatocytrs was 26.2% (16/61),24.6% (15/61) and 44.8% (13/31) respectively.There was no concordance of the existence of HBV-DNA in the serum,peripheral mono-nuclear cells and hepatocytes in an individual.For example,HBV-DNA was absent in the serum but present in mononuclear cells and hepatocytes in 11 cases.In fact,peripheral mononuclsar cells can serve as the reservoir for HBV to replicate.It cannot be denied that HBV can replicate in an individual even though HBV-DNA is negative in the serum.

An Approach to Clinical and Pathological Classification of Chronic Active Hepatitis / 第三军医大学学报

The clinical and pathological data of 208 cases of chronic active hepatitis (CAH) were analyzed, pre liminarily. CAH can be categorized into 6 clinical types: (1) anicteric type, (2) icteric type,(3) severe jaundice and ascites type, ( 4 ) advanced cirrhrotic type, ( 5 ) obstructive jaundice type, and ( 6 ) atypical type.The pathological findings of the 208 cases can be divided into the following. (1) periportal necrotico-inf lammatory type, in which piecemeal necrosis(PN) was prominant, ( 2 ) bridging necrotic type, in which PN and bridging necrosis were present, ( 3 ) submassive necrotic type, multilobular or submassive necrosis occurred, ( 4 ) hepatitis-cirrhotic type, PN with fibrous septa and formation of pseudolobules were seen, and ( 5 ) cholestatic type, PN with intrahepatic cholestasis was pathognomonic.

NECESSITY OF CLINICAL TYPING FOR SUBACUTE AND CHRONIC SEVERE VIRAL HEPATITIS / 解放军医学杂志

According to clinical manifestations, 122 cases of subacute and chronic severe viral hepatitis confirmed by pathologic examination, were divided into two types. The severe jaundice-ascites(SJ-A) type accounted for 93 cases (76.2%) and the subfulminant hepatic failure (SFHF) type 29 cases (23.8%), with a ratio of 3.2 : 1. For SFHF type, hepatic encephalopathy was the first prominent feature and ascites might or might not appeared later. Therefore, this type was most easily misdiagnosed as fulminant hepatitis. In comparison with SJ-A type, the average elevation of serum bilirubin was lower and the average depression of prothrombin activivty more remarkable in SFHF type, suggesting that the degree of hepatic necrosis was more severer and the progress to hepatic failure more rapid. The mortality of SFHF type (93.1%) was markedly higher than that of SJ A type (62.4%) (x2 = 7.488, P