ABSTRACT
Objective To develop a kit of time?resolved fluoroimmunoassay(TRFIA)for detection of Schistosoma japonicum protein SjP38,and evaluate its effectiveness. Methods The anti 9G7 SjP38 monoclonal antibody was used as the capture anti?body coated with 96?hole plate,and the Eu3+labeled 1A6 monoclonal antibody was used as the detection antibody to establish the TRFIA SjP38 kit. In addition,the accuracy,sensitivity,precision,stability and coincidence rate to pathogenic diagnosis of the kit were evaluated. Results This established kit possessed high accuracy,wide linear range from 2 to 1 250 ng/ml,high sensitivity with the minimum detectable concentration of 0.14 ng/ml,and good precision(the coefficient variation of the intra?and inter?assay were 3.6%to 4.6%and 5.1%to 6.7%,respectively). The stability tests showed that the reagents could be stable for six months at 4℃,7 d at 37℃. The positive and negative corresponding rates to the pathogen detection method were 95%and 100%respectively. Conclusion All the performance and detection indicators of the kit have reached the requirements of clinical test,but its clinical application still needs further validation.
ABSTRACT
Objective To investigate the effect of overexpression of miR-30b on the proliferation,cell cycle,apoptosis and invasion of gastric cancer cell line SGC-7901 and AGS,and the inhibitory effect on the tumor formation in vivo.Methods SGC-7901 and AGS cells were transfected with miR-30b mimics and miR-control,and qRT-PCR was used to detect the expression levels of miR-30b.Western blot assay was used to detect the expression of eIFSA2 protein.CCK-8 assay was used to measure the cell proliferation.Flow cytometry was used to analyze cell cycle and apoptosis.Transwell assay was used to detect cell invasion.In addition,the SGC-7901 and AGS cells transfected with miR-30b mimics and miR-control were injected into nude mice to observe the tumor formation and the expression of eIFSA2 protein in vivo.Results Results of qRT-PCR showed that the relative expression of miR-30b was significantly higher than that of miR-control group (P ＜ 0.05).Western blot assay showed that the expression of eIF5A2 protein was decreased in miR-30b mimics group.CCK-8 assay showed that cell proliferation was inhibited in miR-30b mimics group.The result of flow cytometry showed that the cell cycle decreased and the apoptosis increased in miR-30b group.Transwell assay showed that the cell invasion was significantly lower in miR-30b group than that of control group (P ＜ 0.05).Overexpression of miR-30b inhibited the formation of tumor and decreased the expression of eIF5A2 protein in vivo.Conclusion Overexpression of miR-30b inhibits the proliferation,invasion and tumor formation of gastric cancer cells,and reduces the expression of eIF5A2 protein,which provides a potential target for gastric cancer treatment.