1.Application of dyes in polyacrylamide upper gel electrophoresis and im-munoblotting
Weiguang CHEN ; Chenjie YAN ; Zhaojin XU ; Yiting DONG ; Zhichao MAO ; Jiawei SUN ; Wantie WANG ; Linbo YUAN
Chinese Journal of Pathophysiology 2025;41(5):1036-1040
AIM:To investigate the effect of dyes,Remazol BrOrange yellow(RBY)and erythrosine(ERY),on the outcomes of immunoblotting analysis when used for staining the concentrate gel in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE).METHODS:Polyacrylamide gels were divided into five groups:the control group(prepared according to the conventional kit protocol),the RBY-stained group with a final concentration of 0.08 g/L,the RBY-stained group with a final concentration of 0.16 g/L,the ERY-stained group with a final concentration of 0.08 g/L,and the ERY-stained group with a final concentration of 0.8 g/L.Gels were prepared and subjected to electro-phoresis,followed by coomassie brilliant blue staining to visualize protein bands.Subsequently,proteins were transferred to PVDF membranes,which were then blocked,incubated with primary and secondary antibodies,washed,and finally ex-posed for imaging to observe the target protein vinculin bands.RESULTS:Compared with the unstained concentrate gel,the loading wells of the RBY or ERY pre-stained concentrate gel were more clearly visible.Analysis of the gels stained with coomassie brilliant blue after electrophoresis and marker visualization showed no significant different in protein elec-trophoretic mobility between prestained and unstained gels.Comparative analysis of the immunoblotting also indicated that the detection of protein samples transferred to PVDF membranes was unaffected.CONCLUSION:Prestaining concen-trate gels with RBY or ERY can enhance the efficiency of gel-based electrophoresis and immunoblotting analysis.
2.Application of dyes in polyacrylamide upper gel electrophoresis and im-munoblotting
Weiguang CHEN ; Chenjie YAN ; Zhaojin XU ; Yiting DONG ; Zhichao MAO ; Jiawei SUN ; Wantie WANG ; Linbo YUAN
Chinese Journal of Pathophysiology 2025;41(5):1036-1040
AIM:To investigate the effect of dyes,Remazol BrOrange yellow(RBY)and erythrosine(ERY),on the outcomes of immunoblotting analysis when used for staining the concentrate gel in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE).METHODS:Polyacrylamide gels were divided into five groups:the control group(prepared according to the conventional kit protocol),the RBY-stained group with a final concentration of 0.08 g/L,the RBY-stained group with a final concentration of 0.16 g/L,the ERY-stained group with a final concentration of 0.08 g/L,and the ERY-stained group with a final concentration of 0.8 g/L.Gels were prepared and subjected to electro-phoresis,followed by coomassie brilliant blue staining to visualize protein bands.Subsequently,proteins were transferred to PVDF membranes,which were then blocked,incubated with primary and secondary antibodies,washed,and finally ex-posed for imaging to observe the target protein vinculin bands.RESULTS:Compared with the unstained concentrate gel,the loading wells of the RBY or ERY pre-stained concentrate gel were more clearly visible.Analysis of the gels stained with coomassie brilliant blue after electrophoresis and marker visualization showed no significant different in protein elec-trophoretic mobility between prestained and unstained gels.Comparative analysis of the immunoblotting also indicated that the detection of protein samples transferred to PVDF membranes was unaffected.CONCLUSION:Prestaining concen-trate gels with RBY or ERY can enhance the efficiency of gel-based electrophoresis and immunoblotting analysis.
3.Meta-analysis of Clinical Effects of Topical Administration of Phenytoin on Wound Healing
Xiaolin XU ; Zhaojin LEI ; Chuan WANG ; Zhongdong LI ;
China Pharmacist 2017;20(1):113-117
Objective:To evaluate the clinical effects of topical administration of phenytoin ( PHT) on wound healing. Methods:The clinical trials on PHT topically used for wound healing were collected from Cocharne Library and PubMed ( from database establish-ment to May, 2016. Meta-analysis was performed using RevMan 5. 0 software and Stata 12. 0 software. Results:A total of 15 stud-ies involving 1 048 patients were included. Topical PHT treatment was significantly associated with complete healing rate (OR=3. 28, 95%CI:1. 23-8. 75, P=0. 02), production rate of health granulation tissue (OR=2. 18,95%CI:1. 33-3. 59, P=0. 002) and aver-age percentage reduction of wound surface size (SMD=1. 77, 95%CI:0. 53-3. 02, P<0. 000 01). However, heterogeneity existed in complete healing rate and average percentage reduction of wound surface size among the studies. Meta-regression analysis showed that wound types (P=0. 02) and treatment periods(P=0. 08) were associated with the heterogeneity of complete healing rate outcomes, and mean age was associated with the heterogeneity of average percentage reduction of wound surface size(P=0. 07). Conclusion:Meta-analysis suggests that topical PHT treatment has significant positive clinical effect on wound healing. There is heterogeneity among the studies, so topical PHT treatment still should be applied in clinical practice prudently.

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