ABSTRACT
【Objective】 To explore the expression of USP9X in platelets and its effect on platelet function. 【Methods】 The expression of USP9X in human and mouse was evaluated by PCR and Western blot. Platelets from young and old mice were separated and prepared, and the expression of USP9X was detected. USP9X inhibitos were used to assess the regulation of USP9X in platelet function, including aggregation, ATP release and spreading. Platelet lysates were collected in different time points to evaluate the change of phosphorylation of Akt in USP9X inhibitors treated platelets. 【Results】 Both human and mouse platelets expressed USP9X. Compared to the young mice, the old mice showed significantly enhanced expression of USP9X(P<0.05). To assess the effect of USP9X on platelet function, USP9X inhibitor was used to pre-incubate platelets for 30 min and platelet function were examined later. Results showed that USP9X inhibitor significantly decreased platelet activation including aggregation, ATP release and spreading(P<0.05). Compared to the control group, the inhibitor treated group showed a significant decrease in the spreading area after 45 minutes. The Western blot results showed a significant decrease in Akt phosphorylation levels of platelets in the USP9X inhibitor treated group. 【Conclusion】 Both human and mouse platelet express USP9X, and inhibition of USP9X decreased platelet function including aggregation, ATP release and spreading. USP9X can also influence the phosphorylation of Akt. The inhibitor of USP9X may become a potential therapeutic target for thrombosis intervention.
ABSTRACT
Objective · To design an immuno-affinity chromatography device for the separation and detection of syphilis specific antibody, then verify its performance of detection and clinical application. Methods · Affinity filler packed by Treponema pallidum (TP) antigen in affinity chromatography can specifically adsorb TP specific antibody (including IgG and IgM) in samples. After balance, elution and desalination, IgG or IgM gold labeled chromatography strip detects the possibly present syphilis specific IgG or IgM antibody. Twenty cases of syphilis antibodynegative samples and 230 cases of syphilis antibody positive clinical specimens were detected by this chromatography device, and 40 cases were also detected by Western blotting.Results ·The standard operation procedure of the affinity chromatography device was optimized, which could effectively detect the specific IgG and IgM antibody of syphilis. The results of 20 syphilis antibody negative samples were all negative. In 230 syphilis antibody positive cases, the detection results were 2 cases with TP-IgG(-) and TP-IgM(-), 210 cases with TP-IgG(+) and TP-IgM(-),10 cases with TP-IgG(-) and TP-IgM(+), and 2 cases with TPIgG(+)and TP-IgM(+). The detection results of 40 cases were compared with the results detected by Western blotting, among which 2 cases detected by affinity chromatography device were TP-IgG(-) and TP-IgM(-), while the results detected by Western blotting were TP-IgG(+) and TP-IgM(-). But the results of the two methods showed no statistically significant difference (P>0.05). Conclusion · The application of the device in separation and detection of IgG and IgM antibodies against TP pathogens is feasible, and it has important value for further application in clinical diagnosis.
