1.Cytokine detection of tuberculosis patients and study of protective immune mechanism
Jiefan TAN ; Yanya SHEN ; Huijuan FAN
Chinese Journal of Laboratory Medicine 2003;0(07):-
Objective To investigate the roles of the cytokine and chemokine in protective immunity of tuberculosis(TB) patient. Methods The RANTES( CCL5) and MIP1-?( CCL3) content of the Supernatant from PBMC stimulated with IL-4、TNF-?、MIP1-?and LPS in TB patients and normal control subjects was assessed using commercially available ELISA kit. Results The RANTES secretion was increased significantly in both TNF-?plus LPS groups[ (1 731. 86?925. 60) ng/L ] and IFN-?plus LPS [(2 120.78?1 120.72)ng/L] compared with LPS group[ (1 102. 56?873. 44) ng/L] in TB patients (P 0. 05). The MIP1-?secretion in response to IL-4、TNF-?、IFN-?was unchanged significantly in PBMC of TB patients and normal subjects. Conclusion TNF-?、IFN-?and RANTES may play important roles in protective immunity of TB patient
2.Study of the ubiquitination of RNF6 on insulin receptor substrate-1
Li ZHU ; Jiefan TAN ; Liyan ZHOU
International Journal of Laboratory Medicine 2014;(11):1382-1383,1386
Objective To construct an eukaryotic expression vetor of ring finger protein 6 (RNF6)and to investigate its ubiqu-itylation on insulin receptor substrate-1(IRS-1).Methods Human RNF6 coding sequence was amplified by polymerase chain reac-tion (PCR)method with human cDNA as template.The pcDNA3.1-CHA-RNF6 was constructed with routine molecular methods and transfected into hepatocarcinoma cell HepG2.Relative quantification of IRS-1 mRNA was preformed by real-time reverse tran-scription PCR.Western blot was applied to detect the expression levels of IRS-1.Results 48 h after the plasmid carring RNF6 gene transfected HepG2 cells,the mRNA level of ISR-1 gene reduced to 69% of control group.The expression of IRS-1 was markedly lower than control group (P <0.01).Conclusion The expression of IRS-1 is markedly down-regulated in HepG2 and enhancement of the ubiquitylation level of IRS-1 contribute to the disorder in insulin signal transferring.