[Transcriptional inhibitory effect of hepatitis B virus X protein on the expression of p53 tumor suppression gene].

Background: To investigate the transcriptional inhibitory role of hepatitis B virus X protein on the expression of p53 tumor suppression gene.

Methods: The promoter sequence of the p53 tumor suppression gene was identified and amplified by bioinformatics and polymerase chain reaction (PCR). The recombinant reporter gene expression vector pCAT3-p53p was constructed and transfected into the hepatoblastoma cell line HepG2 and cotransfected with pcDNA3.

Thermodynamic study on antibacterial effect of different extracts from Radix Isatis.

Objective: To study and analyze the antibacterial effects of different extracts from Radix Isatis.

Methods: Staphylococcus aureus was used as the studied object in the experiment. Antibacterial effects of extracts from Radix Isatis were observed by thermocalrimetry on Staphylococcus aureus, together with common pharmacological experiments.

Cloning and characterization of a novel hepatitis B virus core binding protein C12.

Aim: To elucidate the biological function of HBV core antigen (HBcAg) on pathogenesis of hepatitis B, a novel gene C12 coding for protein with unknown function interacting with HBcAg in hepatocytes was identified and characterized.

Methods: HBcAg bait plasmid pGBKT7-HBcAg was constructed and transformed into yeast AH109, then the transformed yeast was mated with yeast Y187 containing liver complementary DNA (cDNA) library plasmid in 2XYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-alpha-gal for screening twice.

Study of transactivating effect of pre-S2 protein of hepatitis B virus and cloning of genes transactivated by pre-S2 protein with suppression subtractive hybridization.

Aim: To investigate the transactivating effect of pre-S2 protein of hepatitis B virus (HBV) and construct a subtractive cDNA library of genes transactivated by pre-S2 protein with suppression subtractive hybridization (SSH) technique, and to pave the way for elucidating the pathogenesis of HBV infection.

Methods: pcDNA3.1(-)-pre-S2 containing pre-S2 region of HBV genome was constructed by routine molecular methods.

[Anti-virus effect of hepatocyte stimulating substance (HSS) on duck hepatitis B virus in vivo].

We used one day old Beijing ducklings infected by duck hepatitis B virus as experimental model to observe anti-virus effect of HSS. Beijing ducklings hatched within one day were injected intravenously with duck hepatitis B virus (DHBV). Positive DHBV-DNA in ducklings' sera were detected 7 days later.

[Growth inhibition of human hepatocellular carcinoma xenograft in nude mice by combined treatment with human cytokine-induced killer cells and chemotherapy].

Objective: To compare the inhibitory effects of cytokine-induced killer (CIK) cells alone, chemotherapeutic drug alone, and CIK cells combined with chemotherapeutic drug on the growth of hepatocellular carcinoma (HCC) cells transplanted in nude mice.

Methods: Peripheral blood mononuclear cells (PBMC) collected from five healthy donors by blood cell separator were incubated in vitro to induce CIK cells in the presence of interferon-gamma (IFN-gamma), IL-2 and anti-CD3 monoclonal antibody (mAb). The phenotype of CIK cells was characterized by flow cytometric analysis.

[Identification of immunological effector cells after autologous cytokine-induced killer cells treatment and its clinical implication in hepatocellular carcinoma patients].

Objective: To investigate the alteration of the cellular profiles of T lymphocyte subsets and dendritic cell subsets in peripheral blood of primary hepatocellular carcinoma (HCC) patients after being transfused with autologous cytokine-induced killer cells (CIK) in patients, then to evaluate the clinical efficacy of the immune therapeutic strategy.

Methods: Peripheral blood mononuclear cells (PBMCs) from 13 patients with primary were collected using blood cell separator, and expanded in the fresh AIM-V medium in the presence of cytokine cocktail including interferon-gamma (IFN-gamma), monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2). The phenotypic patterns of CIK cells were longitudinally characterized by flow cytometry on day 0, 4, 7, 10,13 and 15 during the incubation period.

Cloning and analysis of the genomic DNA sequence of augmenter of liver regeneration from rat.

Objective: To search for genomic DNA sequence of the augmenter of liver regeneration (ALR) of rat.

Methods: Polymerase chain reaction (PCR) with specific primers was used to amplify the sequence from the rat genome.

Results: A piece of genomic DNA sequence and a piece of pseudogene of rat ALR were identified.

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.

Synergetic anticancer effect of combined quercetin and recombinant adenoviral vector expressing human wild-type p53, GM-CSF and B7-1 genes on hepatocellular carcinoma cells in vitro.

Aim: This study investigated the anti-cancer effect of combined quercetin and a recombinant adenovirus vector expressing the human p53, GM-CSF and B7-1 genes (designated BB-102) on human hepatocellular carcinoma (HCC) cell lines in vitro.

Methods: The sensitivity of HCC cells to anticancer agents was evaluated by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The viability of cells infected with BB-102 was determined by trypan blue exclusion.

[A preliminary study on the heterogeneity of preS2 region in hepatitis B virus].

Objective: To investigate the mutation of the preS2 gene sequence of HBV and clarify the significance of HBV quasispecies groups in the patients with chronic HBV infection.

Methods: A set of specific primers were synthesized according to HBV DNA sequence of a Chinese strain. The preS2 gene region was amplified by PCR method from the sera of 51 patients with chronic HBV infection, and the PCR products from 5 patients were subcloned into pGEM Teasy vectors.

[The study on heterogeneity of hepatitis B virus DNA].

Objective: To investigate the HBV quasispecies groups in the patients with chronic HBV infection.

Methods: Specific primers ware synthesized according to HBV strain found in China, the preC/C gene, reverse transcriptase region, whole S region, X gene and whole genome ware amplified by PCR method from the serum of 18 patients with chronic HBV infection, and then the PCR products were subcloned into pGEM Teasy vectors. Positive clones with target sequences were selected out for sequencing.

The expression of P21 and P15 proteins in liver tissue of chronic viral hepatitis and hepatocellular carcinoma and their relation to cell apoptosis.

Objective: To study the expression of P21 and P15 proteins in liver tissue of chronic viral hepatitis (CH) and hepatocellular carcinoma (HCC) and their relation to cell apoptosis.

Methods: The expression of P21 and P15 proteins in 36 CH patients liver tissues were detected by immunohistochemistry and the DNA damage of the hepatocytes were detected by TDT mediated dUDT nick and labeling (TUNEL). 10 cases of HCC were also studied.

[Cloning and sequence analysis of human genomic DNA of augmenter of liver regeneration hepatitis].

Objective: To clone the human genomic DNA of augmenter of liver regeneration (ALR) and specify the intron-exon structure.

Methods: Using human ALR cDNA sequence as a reference and BLAST path as a nucleotide homology search tool, GenBank has been searched for ALR homologous genomic DNA sequence. The intron-exon sequences were defined by the Breathnath-Chambon rule.