differences in gene expression profile induced by genotype 1b hepatitis C virus core isolated from liver tumor and adjacent non-tumoral tissue

Core protein of the hepatitis C virus [HCV] has an important role in HCV self-replication, pathogenesis and carcinogenesis. To identify the effect of core proteins from different quasispecies of HCV genotype 1b expressed in a HepG2 cell line on human gene expression profiles. Core protein eukrocytic expression plasmids [pEGFP-N1] containing different quasispecie core protein genes of genotypes 1b HCV derived from of HCV-related hepatocellular carcinoma [HCC] tumoral tissue [T] and non-tumoral tissue [NT] were constructed and then transfected to HepG2 cell line. The gene expressions spectrum in the cell expression core proteins from T and NT were compared with those in the control by Affymetrix human genome HG-U 133 plus 2.0 microarray. Different gene expression profiles were acquired between HepG2 expressing core proteins derived from T and NT tissues. Both core proteins caused the modulation of several genes that are up/down-regulated as compared to control, including genes involved in oncogenesis, signal transduction, cell apoptosis, and cell growth cycle regulation. Surprisingly, only one gene-CNSK1A1- was up-regulated by both T and NT core variants. Core proteins isolated from tumoral or non-tumoral nodules mediate expression of different cellular genes suggesting that variants isolated from different quasispecies may have different biological effects

Influence of fast inhibition to virus replication on immune rejection and prognosis of acute-on-chron-ic livre failur e patients / 中国医师杂志

Objective To analyze the result of treatment for acute-on-chronic liver failure patients with fast high efficiency Nucleoside and to explore the relations among inhibition to virus replication , liver failure development and immune rejection .Methods Sixty-two cases of acute-on-chronic liver failure pa-tients with HBV DNA(+) were divided into study group (treated with a kind of fast and nucleoside , n =30) and control group( n =32).HBV DNA,CD4 +T,CD8 +T, C3,C4 TBIL,PTA were observed at treat-ment 0w,2w and 4w.Results All of the study and control group patients , serum HBV DNA were positive before treated.And the levels of CD4+,CD8 +,C3,C 4,TBIL,PTA of study group were not significantly compared with control group .At treatment 2w , the rate of HBV DNA diverted negative in study group 90.0%(27/30), was significantly more then control group (9.4%, 3/32)(χ2=37.14 , P <0.01).But the CD4 +,CD8 +,C3,C4,TBIL,PTA levels were not significantly however between study and control group . At treatment 4w ,the rate of HBV DNA diverted negative in study group (96.7%, 29/30), was significant-ly more then control group(12.5%,4/32) (χ2 =40.74, P <0.01).CD4 +, CD8 +,C3,C4,TBIL,PTA levels of the study group were significantly more compared with the control group .The CD4 +level of study group (495.33 ±89.91)cells/ml, was higher significantly then control group (270.34 ±97.74)cells/ml( t=9.42, P <0.01),the CD8 +level (571.03 ±120.15 ) cells/ml, was higher significantly then control group(224.88 ±79.68)cells/ml( t =13.45, P <0.01).The C3 level of the study group (0.28 ±0.11) g/L, was lower significantly then control group ( 0.68 ±0.13 ) g/L ( t =13.13 , P <0.01 ) , the C4 level (0.12 ±0.06)g/L, was lower significantly then control group (0.23 ±0.10)g/L( t =4.92, P <0.01). The TBIL level of study group ( 653.93 ±131.02 )μmol/L, was higher significantly then control group (285.63 ±154.63)μmol/L( t =10.09, P <0.01),the PTA level (17.13 ±7.07)%, was lower signifi-cantly then control group(50.94 ±13.68)%( t =12.10, P <0.01).The death rate of the study group( 57.9%) was higher significantly compared with the control group (28.1%)(χ2 =6.39, P <0.05).Con-clusion Treatment of chronic severe hepatitis with fast and high efficiency nucleoside may arise the T cell subset level and make the immune rejection strength , as a result the liver failure maybe far away from cure .

Experimental study on bond strength of machinable composite resin and dentin with three kinds of resin cements / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To examine the shear and microtensile bond strength between a newly developed dental machinable composite resin (polymethylmetacrylate/nano SiO2-ZrO2, PNSZ) and dentin cemented using three resin luting systems and to select the most suitable one.</p><p><b>METHODS</b>The shear and microtensile bond strength between the machinable composite resin and dentin cemented using three resin luting systems (Group A:RelyX ARC, Group B:Panavia-F,Group C:Variolink II) were tested. The broken specimens were observed with a stereomicroscope (x 50) to compare their failure modes. RESULTS; In the shear tests, no significant difference was found in bond strength among Group A [ (14. 07 +/- 4. 67) MPa] ,Group B[ (13.17 +/- 4. 63) MPa] and Group C [ ( 12. 10 +/- 2.18) MPa] (P > 0.05) . In the microtensile tests, no significant difference was found in bond strength among Group A [(11.49 +/- 4.90) MPa],Group B[(9.66 +/- 4.15) MPa].and Group C[(10.11 +/- 4.20) MPa](P > 0. 05).The failure modes of all the three resin cements were predominantly adhesive failures at the dentin/cement interface.</p><p><b>CONCLUSIONS</b>The three types of resin cements showed similar results in bond strength between the dental machinable composite resin and dentin. Bonding at the resin/cement interface was stronger than that at the dentin/cement interface.</p>

Proapoptotic and pronecrosis effect of different truncated hepatitis C virus core proteins / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To study the roles of different truncated hepatitis C virus (HCV) core proteins (CORE) in the pathogenesis of HCV persistent infection and hepatocellular carcinoma (HCC) and to assess intracellular localization in transiently transfected cells.</p><p><b>METHODS</b>Seven truncated GFP (green fluorescent protein)-CORE fusion protein expression plasmids were constructed, which contained HCV CORE sequences derived from tumor tissues (BT) and non-tumor tissues (BNT) from one patient infected with HCV. Amino acid (aa) lengths were BT: 1-172 aa, 1-126 aa, 1-58 aa, 59-126 aa, 127-172 aa; BNT: 1-172 aa and C191: 1-172 aa respectively. Subcellular localization of CORE-GFP was analyzed by con-focal laser scanning microscope. Apoptosis and necrosis were quantified by flow cytometry.</p><p><b>RESULTS</b>Different truncated CORE-GFP localized mainly in the cytoplasm, but nuclear staining was also observed. HCV CORE could induce apoptosis and necrosis, and different truncated COREs could induce cell apoptosis and necrosis at different levels. Among the same length 1-172 aa of BT, BNT and C191, the cell apoptosis and necrosis percentage of BT is highest, and C191 is the lowest (BT>BNT>C191). To the different fragment COREs of BT, N-terminal of CORE induced apoptosis and necrosis higher, compared with that of C-terminal (1-172 aa>1-126 aa>1-58 aa>127-172 aa>59-126 aa).</p><p><b>CONCLUSION</b>These results suggest HCV CORE could induce apoptosis and necrosis of cells, which might play an important role in the pathogenesis of HCV persistent infection and HCC and the different CORE domains of different HCV quasi-species might have some difference in their pathogenesis.</p>

Construction and expression of plasmids containing different truncated core proteins of hepatitis C virus derived from tumor and non-tumor tissues / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the role of different truncated core proteins (CORE) of hepatitis C virus (HCV) played in the pathogenesis of HCV persistent infection and hepatocellular carcinoma, and to construct seven different truncated prokaryotic expression plasmids of HCV CORE.</p><p><b>METHODS</b>The gene sequences of different truncated HCV genotype 1b CORE were amplified from plasmids containing CORE sequences derived from tumor and non-tumor tissues of a patient infected with HCV. Amino acid (aa) lengths of HCV BT (from tumor tissue, patient B) were: 1-172 aa, 1-126 aa, 1-58 aa, 59-126 aa, 127-172 aa; of BNT (non-tumor tissue, patient B) were: 1-172 aa and HCV C191 (HCV-J6): 1-172 aa. PCR products were cleaved with restriction enzymes BamH I and EcoR I and cloned into pGEX-4T-1. Positive clones were transformed into BL21 and glutathione S-transferase(GST)-CORE fusion proteins were expressed with isopropylthio-beta-D-galactoside (IPTG) induction, purified and verified by Western blot.</p><p><b>RESULTS</b>Different truncated GST-CORE fusion proteins were expressed with different quantities. Except the fragment of 59-126 aa, the longer the fragment, the less its expression. The levels of truncated expression of CORE of BT and BNT were higher than that of C191, even though they all contained 1-172 aa. Some of truncated CORE of HCV genotype 1b could form dimmers.</p><p><b>CONCLUSIONS</b>Successful construction of truncated GST-CORE expression plasmids lays a basis for future study of the function of different domains of CORE of different HCV strains; different expression levels of HCV COREs might be related to their different hydrophobicity, cytotoxicity and their functions in HCV structure and their roles played in the pathogenesis; the domain of 59-126 aa is responsible for the HCV genotype 1b CORE dimmer formation.</p>