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OBJECTIVE@#The optimal multiplicity of infection (MOI) of the recombinant adenovirus Ad-Rad50-GFP carrying a mutant Rad50 gene expression region on the cell growth of nasopharyngeal carcinoma and the viral amplification efficiency of CNE1 cell infected by this adenovirus were studied.@*METHOD@#The biological titer of Ad-Rad50-GFP was measured by end point dilution method. The impact of recombinant adenoviral vector transfection on the growth of CNE1 cells was observed by cell growth curve. Transfection efficacy of recombinant adenoviral vector was observed and calculated through fluorescence microscope. The expression f mutant Rad50 in the Ad-Rad50-GFP transfected CNE1 cells with optimal MOI was detected by Western Blot after transfection.@*RESULT@#The biological titer of Ad-Rad50-GFP was 1.26 x 10¹¹ pfu/ml. CNE1 cell growth was not influenced significantly as they were transfected by recombinant adenoviral vector with MOI less than 50. Transfection efficacy of recombinant adenoviral vector was most salient at 24 hours after transfection, with the high expression of mutant Rad50, and the efficiency still remained about 70% after 72 hours.@*CONCLUSION@#Recombinant adenoviral vector Ad-Rad50-GFP could transfect CNE1 cells as well as result in the expression of mutant Rad50 in CNE1 cells effectively. MOI = 50 was the optimal multiplicity of infection of CNE1 cells transfected by recombinant adenoviral vector Ad-Rad50-GFP.
Assuntos
Humanos , Adenoviridae , Carcinoma , Linhagem Celular Tumoral , Vetores Genéticos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas , TransfecçãoRESUMO
Objective To investigate the effects of 17-β estradiol on hepatocyte apoptosis and the expression of Bcl-2 and Bax in hepatic tissue after reduced-size ischemia reperfusion injury and its mechanism in liver protection.Methods A rat model of reduced-size hepatic ischemia-reperfusion injury was established in 75 male Sprague-Dawley rats.They were randomly allocated into three groups:Sham group,ischemia-reperfusion(IR)group,and 17-β estradiol(E2 + IR)group.Liver functions,liver histology and hepatocellular apoptosis rates were observed after reperfusion.Hepatocellular ap optosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)method and the expression of Bcl-2 and Bax were determined by Western blotting.Results The levels of ALT and AST were higher and peaked after 12 h of reperfusion in the IR group compared with the sham group.The histological changes in the liver of the IR group consisted of hepatocyte swelling,hepatic sinusoids narrowing,inflammatory cell infiltration and hepatocyte necrosis in some areas of the livers.The IR group also exhibited an increased rate of hepatocellular apoptosis at 12 h after reperfusion.The protein expression of Bcl-2 decreased while the expression of Bax increased.In the 17-β estradiol group,the levels of ALT and AST were lower,the pathological changes were milder and the rate of hepatocellular apoptosis was lower at 12 h in comparison to those of the IR group.The expression of Bcl-2 was higher and the expression of Bax was lower in the 17-β estradiol group in comparison to those of the IR group.Conclusions 17-β estradiol can relieve the hepatic ischemia reperfusion injury in rat livers.17-β estradiol may inhibit apoptosis in hepatic tissue by up regulating Bcl-2 and down-regulating Bax,thus producing a protective effect on hepatic ischemia-reperfusion injury.
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Objective To investigate the effects of osteogenic growth peptide (OGP) in combination with extracorporeal shock waves (ESWs) on osteoblast proliferation.Methods Passaged cells were divided into four groups for different treatments:a control group,an OGP + ESW group,an ESW group,and an OGP group.After the respective treatments,the cells were cultured for 24 h,48 h and 72 h and counted using methylthiazdy tetrazolium (MTT) and an inverted fluorescence microscope. Immunohistochemical examination was used for detecting protein kinase A (PKA)activity,and a reverse transcription-polymerase chair reaction (RT-PCR) was used for examining PKA mRNA expression at 24 and 48 hours.Results Cell counting revealed that cell proliferation in the OGP + ESW,ESW and OGP groups was significantly promoted compared with the control group.Cell proliferation was greatest in the OGP + ESW group.The immunohistochemical examination showed positive staining intensities in the OGP + ESW,ESW and OGP groups significantly higher than in the control group.The positive staining intensity in the OGP + ESW group was again the highest.PKA activity was also significantly higher in the OGP + ESW,ESW and OGP groups than in the control group with the level in the OGP + ESW group the highest.Conclusion OGP in combination with ESW has a synergistic effect in stimulating osteoblast proliferation and growth.
