1.In vitro antiviral efficacy of mycophenolic acid against hepatitis B virus
Yongfeng YANG ; Deming TAN ; Yutao XIE
Chinese Journal of Infectious Diseases 1997;0(04):-
Objective To evaluate the in vitro inhibitive activities of mycophenolic acid against hepatitis B virus in vitro. Methods The different concentrations of the test drugs in DMEM were added to the well with the monolayer growth of HepG2.2.15 cells. The culture medium was refreshed with DMEM containing the appropriate drugs every 3 days. At 3rd, 6th and 9th days, the supernatant was collected for HBsAg quantitative assay, total RNA was isolated from cells for RT-PCR and realtime-PCR assays. Results It was found no significant anti-HBV effect of mycophenolic acid on concentration of 2~250 ?g/ml, and mini inhibitory efficacy of HBV replication on concentration of 1250 ?g/ml. There was significant anti-HBV effect of lamivudine on concentration of 5 ?g/ml. When combined with lamivudine mycophenolic acid can significantly potentiate anti-HBV activities of lamivudine, and it can get maximal inhibition effect on concentration of 10 ?g/ml. Conclusions Mycophenolic acid could inhibit HBV replication and potentiate anti-HBV activities of lamivudine.
2.Effect of Kang-qian-er-hao on experimental hepatic fibrosis in rats
Ping LI ; Guoling HU ; Deming TAN
Chinese Journal of Infectious Diseases 2001;0(05):-
Objective On the basis of the lasted clinical experience,our group will discuss the treatmented mechanism of Chinese herb kang-xian-er-hao(KXEH) ameliorate hepatic cirrhosis. Methods Male wistar rats were divided into five groups,excepted for normal group N,the remnant four groups were all given intraperitoneal injection of porcine serum((0.5) ml/time,2 times/week,total 12 weeks).In KXEH early treatment group B,the rats were fed with KXEH by gavage,1 g/100 g,once a day at the third week.In KXRH late treatment group C,the rats were fed with KXEH by gavage,1 g/100 g,once a day at the ninth week.In ?-interferon treated group D the rats were subcutaneous injection ?-interferon((0.1) million) every day at the ninth week.The model group A and normal group N were fed with the same amount of saline by gavage.The rats were killed at the end of the twelfth weeks,the formation of liver fibrosis was observed with HE stain and Masson stain.The expression of Smooth muscle actin(SMA) was observed by immunohistochemistry.As well as SMA,collagen Ⅰ、Ⅲ mRNA and Smad3 mRNA,which is TGF-?1 downstream signal,were detected in liver samples with RT-PCR assay. Results In KXEH treated group B and C,the body weight was heavier,the size of liver and spleen was smaller and the ratio of liver weight/body weight and spleen weight/body weight was decreased compared with the model group A(P
3.Effect of STAT3 siRNA on proliferation and apoptosis of gastric cancer cells in vitro
Wei ZHANG ; Qiang WU ; Deming TAN
Journal of Third Military Medical University 2003;0(07):-
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.
4.Effects of truncated type Ⅱ receptor of TGF-?_1 expressing plasmid on rat liver fibrosis
Ling WANG ; Guoling HU ; Deming TAN
Chinese Journal of Infectious Diseases 2001;0(03):-
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
5.Study on the relationship of the disease activity of hepatitis B virus infection and the complexity of S region quasispecies of HBV
Yingxia LIU ; Guoling HU ; Deming TAN
Chinese Journal of Infectious Diseases 2001;0(05):-
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
6.Association between the tumor necrosis factor gene polymorphisms and the clinical types of patients with chronic hepatitis B virus infection
Xuwen XU ; Menghou LU ; Deming TAN
Chinese Journal of Infectious Diseases 2000;0(02):-
Objective To explore whether the polymorphisms within the tumor necrosis factor alpha (TNF-?) and TNF-? gene are associated with the clinical types of patients with chronic hepatitis B virus (HBV) infection. Methods By using a restriction fragment length polymorphism (RFLP) assay of polymerase chain reaction (PCR) products, the single nucleotide polymorphisms (SNPs) within TNF-? and TNF-? gene among 56 patients with chronic severe hepatitis B and 71 patients with chronic mild hepatitis B or asymptomatic carriers as well as 90 healthy controls were analyzed. The TNF-? concentration of serum in 56 patients with chronic severe hepatitis B and 30 healthy controls were determined by radio immunity assay (RIA). Results The frequencies of the TNF1/2 genotype and the TNF2 allele were significantly increased in patients with chronic severe hepatitis compared with healthy controls (25% vs 11.1%,P=0.028;12.5% vs 5.6%,P=0.036) and patients with chronic mild hepatitis B and asymptomatic carriers (25% vs 8.5%,P=0.011;12.5% vs 4.2%,P=0.015). Furthermore, heterozygotes carrying TNF2 allele had significantly higher levels of serum TNF-? than homozygotes for the wild type allele among all patients with chronic severe hepatitis B (P
7.The affection of neutralizing antibody on therapeutic effect of IFN?-2b
Guozhen LIU ; Guoling HU ; Deming TAN ;
Chinese Journal of Infectious Diseases 1997;0(04):-
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
8.Hepatitis B virus X protein induces expression of fibrosis-related factors in human hepatic stellate cells
Guanghui GUO ; Deming TAN ; Pingan ZHU
Journal of Chinese Physician 2009;11(8):1032-1035
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.
9.Clinical significance of serum amyloid A protein in patients with chronic hepatitis C virus infection
Lin SONG ; Deming TAN ; Zhixiong FANG
Chinese Journal of Clinical Infectious Diseases 2008;1(2):96-98
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.
10.A Study on the Effects of Interferon-alpha on Hepatitis B virus in Vitro
Yongfeng YANG ; Deming TAN ; Yutao XIE
Journal of Chinese Physician 2001;0(05):-
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.