f fibrosis-related factors in LX-2cells were significantly increased after co-cultured with QSG7701-HBx cells, which proved that HBx could induce fibrogenesis in vitro.

Objective To investigate the clinical significance of serum amyloid A protein in patients with chronic hepatitis C (HCV) infection. Methods Serum amyloid A (SAA) levels were detected by ELISA in 131 patients with HCV infection and 20 normal controls. The expression of SAA-mRNA in peripheral blood mononuclear cells (PBMC) was detected by RT-PCR from some blood samples of HCV patients and normal controls. Results The SAA levels in the patients with chronic HCV infection were markedly higher than those in normal controls (t = 17. 14, P < 0. 01 ). The expression of SAA-mRNA detected by RT-PCR was closely correlated with concentrations of SAA measured by ELISA ( r = 0.86, P <0.01 ). No correlation was found between SAA expression and serum HCV RNA titers, as well as between SAA and serum ALT in patients with chronic HCV infection. Conclusion SAA levels are increased in patients with chronic HCV infection, which is not correlated with HCV RNA titers and serum ALT levels.

Objective To evaluate the in vitro inhibiting activities of interferon-alpha (INF-?) against hepatitis B virus (HBV). Methods HepG2 2 2 15 cells were treated with INF-?. At 3rd, 6th and 9th days after treatment, the supernatant was collected for HBsAg quantitative assay, and total RNA of the cells was isolated for RT-PCR and realtime-PCR assay of HBV mRNA. Results There were no significant differences in HBsAg titer and virus mRNA level at 3rd and 6th days between the experimental and control groups. At 9th day, HBsAg titer and virus mRNA level in INF-? group were significantly lower than those in control group. And 500IU/ml INF-? could get maximal inhibiting effect against HBV. Conclusion INF-? can inhibit transcription and expression of HBV gene in HepG2 2 2 15 cells, which indicated that INF-? can inhibit HBV replication directly.

Objective To design and construct a vector of signal transducer and activator of transcription 3 (STAT3) small interfering RNA (siRNA),and to investigate the effect of the recombinant plasmid on the proliferation and apoptosis of gastric cancer cell SGC7901. Methods The PAVU6+27-STAT3 siRNA expression vector was constructed, then transfected into the cultured gastric cancer cells SGC7901 by DOTAP method. STAT3 mRNA and protein in SGC7901 were determined by RT-PCR and Western blotting respectively. Cell proliferation was tested by MTT and 3H-TdR incorporation. Cell apoptosis percentage and bcl-2 expression were observed by TUNEL and flow cytometry respectively. Results The PAVU6+27-STAT3 siRNA expression vector was successfully constructed. In the transfected SGC7901 cells, cell proliferation, 3H-TdR incorporation and bcl-2 expression were decreased, and cell apoptosis percentage was increased. Conclusion PAVU6+27-STAT3 siRNA expression vector may be an efficient method to inhibit the proliferation and promote the apoptosis of gastric cancer cells.

Objective To observe the effects of truncated type Ⅱ receptor of TGF ? 1 expressing plasmid on rat liver fibrosis. Methods An eukaryotic expression plasmid of a truncated type Ⅱ receptor of TGF ? 1 was constructed by recombinant DNA technique. Sequence analysis of this recombinant plasmid revealed that it contained the complete extracellular and transmembrane domains of TGF ? 1 Ⅱ receptor but lacked most of the cytoplasmic kinase domain. The recombinant plasmid encapsulated by liposome was transferred into the rats by intraperitoneal injection and its effects on the grade of the pathologic fibrosis and the levels of HA,LN,PCⅢ,Ⅳ C were observed. Results The levels of serum HA,LN,PCⅢ,Ⅳ C were lower in the early treatment group than those in the model control group ( P 0.05). Either in the early or later treatment group, the grade of liver fibrosis was decreased apparently compared with the model control group ( P

Objective To study the complexity of S region qusispecies in various disease stages of chronic hepatitis B virus(HBV) infection and its relation to disease activity. Methods Serum samples were obtained from 112 patients with chronic hepatitis B virus infection;22 with chronic carries(ASC),30 with chronic mild or moderate hepatitis(CH),60 with fulminant hepatitis failure(FHF). HBV qusispecies populations were separated by the single strand conformation polymorphism (SSCP) method targeted the S region and DNA sequencing analysis. Results The number of SSCP bands detected in the patients with ASC、CH and FHF was 1.45?0.13,3.70?0.22 and 5.93?0.24, respectively. There was a statistically significant difference in the number of quasispecies among various disease stages ( P

In order to study the affection of neutralizing antibody on the therapeutic effect of IFN? -2b,NBA was used to detect the neutralizing antibody to against IFN in the sera from 27 patients with chronic hepatitis B.The results indicated that neutralizing antibody against IFN was present in 15 of 27 patients who had been treated with IFN?-2b,and the overall frequency of neutralizing antibody was 55.6%(15/27).At the end of IFN therapy,HBV DNA was undetectable in 4 of the 15 patients(4/15, 26.7%)with neutralizing antibody.By contrast.HBV DNA became undetectable in 10 of the 12 patients (10/12,83.3 % ) without neutralizing antibody (P

Objective To construct an eukaryotic expression vector of human sTNFR1 and to investigate its inhibitory effects to the bioactivity of TNF-?.Methods The total RNA was extracted from HeLa cells and used as a template to amplify human sTNFR1 gene by reverse transcription polymerase chain reaction(RT-PCR).The PCR products were cloned into T vector and sub-cloned into vector pcDNA3.1(-),an eukaryotic expression vector.The recombinant plasmid pcDNA3.1(-)-sTNFR1 was transfected into QSG7701 cells by using lipofectamine,RT-PCR was performed to detect the expression of sTNFR1,MTT was used to observe sTNFR1 gene 's inhibitory effect on TNF-?.Results QSG7701 has a higer expression level of sTNFR1 mRNA than pcDNA3.1(-) trancfected control.The cytotoxic effect of TNF-? was inhibited to the extent of 64.8% when its concentration was 100 ?g/L.Conclusion We constructed the eukaryotic expression vector containing human sTNFR1 gene and the cytotoxicity of TNF-? is inhibited in pcDNA3.1(-)-sTNFR1/QSG7701 cells.

Objective To establish a multiplex polymerase chain reaction (M-PCR) assay to detect HBV DNA in the peripheral blood mononuclear cells(PBMCs) of chronic HB patients. Methods One pair of primer amplifying HBV genome DNA and another pair of primer amplifying HBV covalently closed circular DNA (cccDNA ) were added to one PCR reaction to detect HBV DNA in PBMCs. Results Various forms of HBVDNA including total DNA and cccDNA could be amplified simultaneously. Among the 30 chronic HB patients, both the HBVDNA and HBVcccDNA in the PBMCs of 23 patients were detected, the positive rate of which was 76.6%. The positive rate of HBV cccDNA accounted for 82.1% of total HBV DNA positive rate. Conclusion HBVDNA in the PBMCs could partially replicate. The M-PCR was successfully set up to amplify HBV genome DNA and HBV cccDNA simultaneously.

Objective To construct the recombinant plasmid carrying human sTNFR1 cDNA, and express sTNFR1 in E. Coli JM109. Methods Total RNA was extracted from Hela cells, and used as a template to amplify human sTNFR1 cDNA by RT-PCR. The PCR products were cloned into T vector, and then sTNFR1 cDNA fragment was subcloned into a prokaryotic expression plasmid pMAL-c2x. The recombinant plasmid was transferred into E. Coli JM109, and induced by IPTG to express fusion protein sTNFR1-MBP. sTNFR1-MBP was purified by amylose resin affinity chromatography(ARAC), and analyzed by SDS-PAGE. Results A 558 bp human sTNFR1 cDNA was amplified by RT-PCR, and successfully inserted into plasmid pMAL-c2x. sTNFR1-MBP was produced in E.Coli after IPTG induction, and a 66 KD sTNFR1-MBP was purified by ARAC. [WTHZ]Conclusion Recombinant plasmid carrying human sTNFR1 cDNA was successfully constructed and epxressed in E. Coli JM109.