Heat-shock protein 27 linked to multi-drug resistance in human hepatic cancer cell lines / 中华肝脏病杂志

<p><b>OBJECTIVE</b>A proteomics approach was applied for finding multi-drug resistance (MDR) related proteins in hepatic cancer cell lines.</p><p><b>METHODS</b>Two-dimensional electrophoresis (2-DE) was used to separate the total proteins of vincristine-resistant hepatic cancer cell line HepG2/VCR and its counterpart HepG2. The differential expression proteins between the two cell lines were identified by both MALDI-TOF-MS and ESI-Q-TOF-MS. Heat-shock protein 27 (HSP27), one of the differential expression proteins in the development of MDR of HepG2/VCR, was analyzed by HSP27 antisense oligonucleotides (ASOs).</p><p><b>RESULTS</b>As a high expression protein in HepG2/VCR, HSP27 was identified, and the suppression of HSP27 expression by HSP27 ASO enhanced the vincristine chemosensitivity of HepG2/VCR (P less than 0.05).</p><p><b>CONCLUSION</b>HSP27 is linked to MDR in human hepatic cancer cell line HepG2/VCR.</p>

Study on quasispecies of hepatitis B virus and the clinical manifestation of their infection by melt curve analysis / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To study the relationship between quasispecies of hepatitis B virus and the clinical manifestations of their infection, and to find the answer of why different quasispecies of HBV with the same genotype can induce different clinical situations.</p><p><b>METHODS</b>Sixty serum samples, in all of which HBVs of genotype B exist, taken from 32 chronic asymptomatic carriers and 28 severe chronic hepatitis patients, were collected to detect quasispecies of HBV DNA by melt curve approach. Then the relationship between quasispecies of HBV of the same genotype and the clinical situation of their infection was studied by comparing the wave crests of the two sample groups.</p><p><b>RESULTS</b>The data of the 60 serum samples of HBV of genotype B detected by melt curve showed that HBV DNA in severe chronic hepatitis patients had more wave crests than that in chronic asymptomatic carriers (P < 0.05), suggesting that HBV in severe chronic hepatitis patients had more quasispecies than in the chronic asymptomatic carriers.</p><p><b>CONCLUSION</b>The numbers of quasispecies of HBV correlate with the clinical situations of their infection. In the patients infected by HBV of the same genotype, those who have more HBV quasispecies would have more severe clinical manifestations.</p>

Cloning , Expression of Human sTNFR1 Gene and the Biological Activity of Its Recombinant Protein / 中国生物工程杂志

Human sTNFR1 (soluble tumor necrosis factor receptor 1) gene was amplified by RT-PCR from Hela cells. A recombinant expression vector of sTNFR1-MBP was constructed in pMAL-c2x, and transformed into E. Coli JM109.It was sequenced and confirmed to be identifical to the sTNFR1 gene in data bank. Recombinant protein sTNFR1-MBP was induced by IPTG and purified by Amylose resin Affinity Chromatography. sTNFR1-MBP was binded to sTNFR1's antibody in Western-blotting. From MTT assays, the results showed that sTNFR1-MBP could effectively block the cytotoxicity mediated by TNF?on QSG7701 cells. Annexin V-FITC staining and flowcytometry were used to observe the recombinant protein's anti-apoptosis capacity and the recombinant protein has marked anti-apoptosis effect in vitro.sTNFR1-MBP had good biological activity and it will be employed in further study.

Effect of diazepam and modafinil on acute hepatic failure in mice / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of diazepam and modafinil on acute hepatic failure in mice.@*METHODS@#Acute liver failure was induced in male Kunming strain mice by enterocoelia injecting the mice with D-GalN and LPS . The mice in the treatment groups were given corresponding drug 2 h before the administration of D-GalN and LPS, and the mice in the control group were given the same dose of distilled water. The 24-hour survival rate, serum alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels were compared. Serum levels of TNF-alpha and IL-1 and the levels of SOD, MDA, GR, GSH, NO and NOS in the liver were determined.@*RESULTS@#Treatment with diazepam increased the survival rate and improved liver histological feature. Diazepam inhibited the serum levels of ALT, AST, TNF-alpha and IL-1, and reduced levels of MDA, NO and NOS and increased levels of GR and SOD in the liver. Modafinil decreased liver histological feature, increased the serum levels of ALT, AST, TNF-alpha and IL-1, increased level of MDA, and inhabited levels of SOD and GR in the liver.@*CONCLUSION@#Treatment with diazepam may suppress the D-GalN/LPS-induced acute hepatic failure and modafinil may facilitate the acute hepatic failure.