Expression and clinical significance of GFRα3 in esophagus squamous cell carcinoma / 临床与实验病理学杂志

Purpose To investgate the expression and clinical significance of GFRα3 in esophagus squamous cell carcinoma (ESCC).Methods Expression of GFRα3 in 114 esophagus squamous cell carcinoma,17 cases of high grade intraepithelial neoplasia and Tis,25 cases nomal tissue was detected by immunohistochemistry.Expression of GFRα3 in 12 paired flesh tissues had also been determined by Western-blot.Results The positive expression of GFRα3 in nomal esophageal tissues,high grade intraepithelial tissues and Tis was 0,29.4％,76.3％ respectively by immunohistochemistry.Expression of GFRα3 in carcinomatous tissues was higher than that of adjacent tissues (P ＜ 0.05).Moreover,the expression of GFRα3 was significantly associated with depth of invasion,higher clinical stage and survival outcome of patients with ESCC.Conclusion The expression of GFRα3 maybe a useful predictor of disease depth of invasion,progression and outcome in ESCC.

Inhibiting effects of three components of Astragalus membranaceus on oxidative stress in Chang Liver cells / 中国中药杂志

The main objective of this research is to investigate the effects of astragaloside IV, calycosin separately glucoside, formononetin on oxidative stress in Chang Liver cells induced by H2O2. In the experiments, Chang Liver cells (a kind of normal human hepatocytes) were used as the research object, bifendate which has a clear hepatoprotective effect was used as the positive control drug, then the oxidative damage model of Chang Liver cells were established by H2O2. Cells were divided into six groups: blank control group, oxidative stress group, astragaloside IV group, calycosin separately glucoside group, formononetin group and positive control group. Then endogenous antioxidant system related indexes were detected by micro plate and colorimetric method; intracellular reactive oxygen species (ROS) were detected by DCFH-DA fluorescent probe; and the expressions of CYP2E1 were evaluated by liver microsomes, mRNA, and protein, respectively with spectrophotometry, Real-time PCR method, and Western blot technique. Results showed that H2O2 decreased antioxidant activity, and increased ROS level and expression of CYP2E1. The above oxidative stress status had been changed with protections of the three components of Astragalus membranaceus (compared with oxidative stress group, P < 0.05, P < 0.01), which taken as a whole had equivalent effects as the drug of positive control group( bifendate). Taken together, three Astragalus membranaceus ingredients all had significant or extremely significant inhibiting effects on oxidative damaged Chang Liver cells which were induced by H2O2, and the oxidative damage of Chang Liver cells had been relieved.

Protective effect of astragaloside IV on oxidative damages of chang liver cell induced by ethanol and H2O2 / 中国中药杂志

<p><b>OBJECTIVE</b>To study the protective effect of astragaloside IV on oxidative damages of Chang Liver cells induced by ethanol and H2O2.</p><p><b>METHOD</b>The alcoholic and nonalcoholic oxidative damage models were established on Chang Liver cells with ethanol and H2O2, respectively. The cells viabilities were detected by MTT assay, transaminase activity and antioxidant ability were detected by micro plate and colorimetric method, reactive oxide species (ROS) was detected by DCFH-DA fluorescent probe and cell cycle was detected by flow cytometry. DNA ladder method was used to detect apoptosis.</p><p><b>RESULT</b>Both kinds of oxidative damage could decrease the viability and antioxidant enzyme activity of Chang Liver cells, and increase the transaminase activity and MDA content of extracellular fluid. The protective effects of astragaloside IV against those two kinds of oxidative damages were significant or extremely significant. Meanwhile, ethanol could decline the level of ROS significantly in the damaged cells, while H2O2 could increase it significantly. And the effect of astragaloside IV was to make ROS return to the normal level. Retardation of cell cycle progression of Chang Liver cells in G0/G1 induced by ethanol or H2O2 was relieved, and apoptosis was also inhibited.</p><p><b>CONCLUSION</b>Astragaloside IV had protective effect on oxidative damages of Chang Liver cells induced by ethanol and H2O2.</p>

Gene sequence analysis and prokaryotic expression of sigmaB protein of Muscovy duck reovirus YB strain / 病毒学报

Muscovy ducks reovirus (DRV) is an important pathogen with a high mortality rate in Muscovy ducks, the researches in the test and the immunity were useful for the prevention and control of DRV infection. In this study, the S3 genes of the three Fujian DRVs were cloned by RT-PCR and sequencing technology. It was found that DRV-YH and YJL were close to avian reovirus (ARV) in the genetic distance, with high identities ranged from 94. 6% to 98. 9%, however, the identities of DRV-YB strain and reference ARV strains in the S3 gene were only 60.6% - 61.7%. The expression vector pET-30a-S3 harboring DRV YB strain S3 gene was constructed and transformed into E. coli BL21, and then the fusion sigmaB protein expression was induced with IPTG. The SDS-PAGE of the expressed products indicated that the fusion protein of approximately 42ku in molecular weight was expressed highly in inclusion body, and made up 67. 7% of the total proteins. The most efficient concentration of IPTG and inducing time were 0. 1 mM and 5h respectively, while the best temperature for expression was 37 degrees C. After purification with the Ni2+ affinity chromatography, the fusion sigmaB protein was 93% of the total proteins, and the purified protein amounted to 0. 86g/L. The Western blot analysis showed that the fusion aB protein was recognized specifically by the antiserum against DRV, confirming that the recombinant fusion protein had good immunoreactivity.