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Objective To investigate the effect of simvastatin on gene expression of L-type calcium channels(LCCs) of myocardial cells in BALB/c mice infected by coxsackievirus B3(CVB3) so as to study the therapeutic effect of simvastatin on viral myocarditis.Methods Sixty male BALB/c mice were divided into five groups randomly(n=12).Mice in viral control group and three groups of oral administration of simvastatin(10,30 and 90 mg?kg-1)were inoculated intrapritoneally with 0.2 mL of CVB3(Nancy strain).Mice in normal control group were inoculated intrapritoneally with 0.2 mL Eagle's solution.The heart samples of all the mice were obtained for hispathological study and detection of myocardial LCCs alpha1 subunits mRNA expression by semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR).Results In viral control group,the mononuclear inflamematory infiltrate was focal or diffuse in myocardium of mice,severe hearts revealed a large area of myocardial necrosis.The degree of inflammatory cell infiltrate and area of necrosis were significantly less in simvastatin groups as compared with viral control group.The myocardial LCCs alpha1 subunits mRNA expression by semi-quantitative RT-PCR in normal control group was much lower than that in viral control group(0.06?0.01 vs 1.37?0.32,P
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Objective:To set up an effective and simple purification method to obtain highly purified prokaryotic protein of PDCD5 and study its stability. Methods:Recombinant PDCD5 protein expressed in E. coli was accumulated as an inclusion body. After washing, the inclusion body was denatured, renatured, digested with thrombine and then purified by two steps of chromatography. The purity of the products was analyzed by capillary electrophoresis and the stability was identified by SDS-PAGE. Results:Capillary electrophoresis showed that the purity of protein was 100%, and molecular weight was 15 800 with pI 5.9. Further bioactivity assay indicated that the purified PDCD5 could enhance the apoptosis of HL 60 cells withdrawing cytokine, which was in a dose dependent manner. Stability analysis showed that the PDCD5 protein was sensitive to temperature and easy to degrade at 4 ℃ and 25 ℃. However, it was relatively stable at -20 ℃ or lyophilized. Conclusion:Highly purified and stable recombinant PDCD5 protein was obtained, which lays a foundation for the functional study and application investigation of PDCD5 .
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Object To develop the gene resources of Jilin Shuangyang sika deer,a rare animal in China and reveal the pharmacological functions of deer medicinal material on the molecular level. Methods A cDNA plasmid library of spleen cells derived from Shuangyang sika deer was set up. Some housekeeping genes were cloned and analyzed. Results One of the cDNA sequences and it's deduced amino acid sequence in deer share high homology with a group of cDNA sequences encoding ribosomal protein L27 in human,rat,Canis and mouse and their corresponding amino acid sequence respectively. It has the completed open reading frame as well. Conclusion A novel full-length cDNA encoding Shuangyang sika deer ribosomal protein L27 is discovered. This sequence has been deposited in Genbank under accession number AF373231.