Comparison between the quantitative and semi-quantitative fluorescence RT-PCR assays for the detection of XDH/XO mRNA expression in different organ tissues of rhesus monkey / 中国比较医学杂志

Objective To analyze the differences between the semi-quantitative RT-PCR and real time quantitative fluorescence RT-PCR assays for detecting XDH/XO mRNA expression in various organ tissues of rhesus monkey, and provide useful reference in methodology of experimental studies.Total RNA was extracted from the myocardium, kidney, testis, skin, and liver tissues, respectively, for detecting XDH/XO mRNA expression in rhesus monkey by semi-quantitative RT-PCR and real time quantitative fluorescence RT-PCR assays.The sensitivity and specificity of the two assays were compared with each other using the same primer sequences and reference genes.Results The expression of XDH/XO mRNA in different organ tissues were detected by both the two PCR assays.The sensitivity of quantitative fluorescence real-time RT-PCR for the XDH/XO mRNA expression in the liver tissue was 39 times higher than that by semi-quantitative RT-PCR.Conclusions Both the quantitative and semi-quantitative fluorescence RT-PCR assays can be used to detect the expression of XDH/XO mRNA in different organ tissues of rhesus monkey.The sensitivity of quantitative fluorescence real-time PCR assay is more sensitive than that of the semi-quantitative RT-PCR assay.

Effects of ginsenoside Rb1 on cell proliferation,differentiation and OPG/RANKL mRNA expression of rat osteoblast cultured in vitro / 中华中医药杂志

Objective:To study the effects of ginsenoside Rb1 on cell proliferation,differentiation and OPG/RANKL mRNA expression of rat osteoblast cultured in vitro.Methods:Rat osteoblast cultured in vitro was treated by a series of different concentration of ginsenoside Rb1 for different time respectively.The proliferation of osteoblast was detected by MTT method.The differentiation of osteoblast was detected by the activity of ALP.mRNA expression of osteoprotegerin (OPG) and RANKL(receptor activator nuclear factor-kappa B ligand,RANK) were semi-quantified by RT-PCR,and the ratio of OPG/RANKL mRNA was used to analyze the differentiation of ginsenoside Rb1 to osteoclast.Results:Ginsenoside Rb1 at a concentration of 1.12?10-9mol/L to 1.12?10-5mol/L can promote the proliferation of osteoblast after 72h of treatment (P