ABSTRACT
<p><b>OBJECTIVE</b>To establish a canine cell line with p53 gene knockdown by lentivirus- mediated RNA interference (RNAi).</p><p><b>METHODS</b>Four pairs of oligonucleotide sequences of p53 gene were synthesized and cloned into the pGMLV-SC5 RNAi vector. Following confirmation by PCR and DNA sequencing, the recombinant pGMLV-p53 plasmids and packaging mix vectors were packaged into mature lentivirus to infect 293T cells. The supernatant of the infected cells was harvested to infect WRD cells, in which p53 expression was detected using Western blotting and real-time PCR. The lentivirus with the strongest p53⁻ silencing effect was packaged for infecting WRD cells at the optimal multiplicity of infection (MOI) to establish a stably infected cell line (WRD/p53⁻) screened using puromycin.</p><p><b>RESULTS</b>The lentivirus carrying p53 shRNA was constructed successfully and a virus titer of 1 × 10⁹ TU/ml. The 4 pGMLV-p53 plasmids all produced strong p53 interference affects, and pGMLV- p53A1 with the strongest effect. The stably infected cells line WRD/p53⁻ was established successfully using pGMLV-p53A1 plasmid.</p><p><b>CONCLUSION</b>The canine cell line WRD/p53⁻ with stable lentivirus-mediated p53 silencing has been established successfully.</p>
Subject(s)
Animals , Dogs , Cell Line , Gene Knockdown Techniques , Genes, p53 , Genetic Vectors , Lentivirus , Plasmids , RNA Interference , RNA, Small Interfering , Real-Time Polymerase Chain ReactionABSTRACT
Objective To investigate the effect of 6 sigma nursing quality management model in rescue of patients with severe trauma.Methods According to the admission sequence,268 patients with severe trauma were selected as control group and treated with traditional first aid pattern; another 292 with severe trauma were chosen as the observation group and treated with 6 sigma nursing management mode(definition,measure,analysis,improvement and control).The two groups were compared in terms of rescue success rate and effective rescue time.Result The rescue success rate in the observation group was significantly higher than that in the control group and the effective rescue time in the observation group was significantly shorter than that in control group(P<0.05). Conclusion The application of 6 sigma nursing management mode in rescue of patients with severe trauma can improve the success rate,shorten the effective rescue time,so it enhances the nursing quality of severe trauma.
ABSTRACT
m-Nisoldipine ( m-Nis ) , isobutyl methyl 1 , 4-dihydro - 2, 6-dimethyl-4-( 3-nitrophenyl )-3 , 5-pyridine dicarboxylate, is a new calcium antagonist. Its effects on hemodynamics and regional blood flow as well as the distribution of cardiac output were evaluated in comparison with Nis with radio-biomicrospheres in conscious rabbits. ( 1 ) m-Nis ( 1 ?g/kg ? min-1 ? 10 min, iv ) increased cardiac output and cardiac index significantly (P0.05) ; ( 2 ) m-Nis increased stroke volume and left ventricular systolic work ( per beat ) while Nis slightly increased the former and decreased the later, and these differences were statistically significant (P