Construction and identification of tetracycline-inducible rat Smad7 eukaryotic expression vector / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a tetracycline-inducible eukaryotic expression vector of rat Smad7.</p><p><b>METHODS</b>The total RNA was extracted from normal rat kidney with Trizol agent. Rat Smad7 cDNA fragment was cloned by RT-PCR, and was inserted into the restriction site between Nhe I and Hind III of the inducible eukaryotic expression vector pBI-L by tetracycline. pBI-L-Smad7 was constructed by digestion and ligation, and detected by restriction endonuclease digestion and sequencing.</p><p><b>RESULTS</b>The recombinant eukaryotic expression vector pBI-L-Smad7 was constructed correctly as confirmed by restriction endonuclease digestion and sequencing. The fragment of pBI-L-Smad7 digested with restriction endonucleases and the sequence of inserted Smad7 cDNA were consistent with the results of theoretical analysis.</p><p><b>CONCLUSION</b>The tetracycline- inducible eukaryotic expression vector of rat Smad7, pBI-L-Smad7, is constructed successfully, which may facilitate further clinical study of Smad7 gene therapy for tissue and organ fibrosis.</p>

Construction and identification of tetracycline-inducible human hepatocyte growth factor eukaryotic expression vector / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a tetracycline-inducible eukaryotic expression vector containing human hepatocyte growth factor (HGF) cDNA.</p><p><b>METHODS</b>Human HGF cDNA fragment was obtained by PCR from pUC-SRalpha/HGF plasmid and inserted into the restriction site between Mlu I and Sal I of the tetracycline-inducible eukaryotic expression vector pBI-L. pBI-L-HGF was constructed by DNA recombination in vitro, and was identified by restriction endonucleases digestion and sequencing.</p><p><b>RESULTS</b>The fragment of pBI-L-HGF digested with restriction endonucleases well corresponded to expectation, and the sequence of inserted HGF cDNA was correct according to the GenBank.</p><p><b>CONCLUSION</b>The tetracycline-inducible eukaryotic expression vector of human HGF pBI-L-HGF has been constructed successfully, which allows further study of HGF gene therapy with much safety and easy manipulation.</p>

Effects of components isolated from Astragalus mongholicus on expression of p-selectin in shock wave induced kidney injury in rabbit model / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effective components of Astragalus mongholicus and their mechanisms in alleviating extracorporeal shock wave lithotripsy (ESWL) induced kidney injury.</p><p><b>METHOD</b>69 male rabbits were randomly assigned into control group, sham treatment group, Total Saponins of Astragalus (TSA) group, Total Flavonoids of Astragalus (TFA) group, and Total Polysaccharide of Astragalus group (TPA). The shock wave treated kidneys were observed for the expression of p-selectin and the change of cells ultrastructure pre and post ESWL. The concentration of Maleic Dialdehyde (MDA) as well as activity of superoxide dismutase (SOD) in the kidney tissues was also analyzed.</p><p><b>RESULT</b>After application of shock wave treatment, p-selectin was expressed extensively in renal glomerulus, renal tubules and renal interstitium of the treated kidneys. It showed a significant increase of MDA levels and decrease of SOD activity in control group (P < 0.05) after ESWL. The comparison between controls and TSA group demonstrated that TSA could significantly reduce the positive rate of p-selectin P < 0.05) and alleviate the injuries of cells ultrastructure. The results also showed a significant decrease of MDA levels and increase of SOD activity in TSA group compared to controls (P < 0.05). The protective effects of TFA and TPA in alleviating kidney injury induced by shock wave were lower than those of TSA; they had no effects of the expression of P-selectin.</p><p><b>CONCLUSION</b>TSA is the main components of A. mongholicus in alleviating shock wave induced kidney injury not only by scavenging oxygen free radicals but also inhibiting the expression of p-selectin.</p>