0.05) after irradiation with low dose of UVB. When the dose of UVB reached 60 mJ/cm2, the mRNA and protein expression of both p21 and Bax increased significantly (both P

Aim To assay the possible targets of adriamycin (ADM), screening ADM resistance related proteins.Methods The drug sensitivity of the cells was analyzed by IC50 assay;RT-PCR assay was used to detect the expression of genes in the cells;CMPK1 protein expression was tested by Western blot assay;the expression of CMPK1 in the cells was decreased by siRNA of CMPK1.Results Data from IC50 assay showed the sensitivity of cells transfected with CMPK1 was increased most(IC50 HEK293-CMPK /IC50 HEK293-Control=0.15, P<0.01), and the expression of CMPK1 protein in ADM resistant breast cells (MCF7/ADM) was lower than that in parent MCF7 cells (P<0.05).When the expression level of CMPK1 was decreased by CMPK1 siRNA, the sensitivity of MCF7 cells to ADM decreased (IC50 MCF7-siCMPK1/IC50MCF7-Control=3.6, P< 0.01), and the sensitivity of MCF7 cells to paclitaxel and gemcitabine also decreased.Conclusions CMPK1 was related to the multidrug resistance of cells, and the expression of CMPK1 was positively related to the sensitivity to drugs, which provides the possibility of CMPK1 as a target in the treatment of multidrug resistance.

Objective To identify the imbalance of T cell-specific transcription factors T-bet/GATA-3,and to explore the modulation with dexamethasone and imiquimod in CD4+T cells from ovalbumin (OVA)sensitized mice.Methods CD4+T cells were obtained fromsingled-cell suspension of spleen(after lysis of RBC).ELISA assay was used to detect the concentrations of IL-4,IL-5 and IFN-γin superna tants and cell pellets,and the expression of T-bet and GATA-3 was detected by Western blot.Resuits In the control group,tIle low levels of IFN-γ were detected in the supernatants during 24 h.In OVA treatment group,the concentrations of IL-4,IL-5 were increased significantly,and the concentrations of IFN-γ were always low in the supernatants.In the dexamethasone treatment group,the concentrations of IFN-γ,IL-4 and IL-5 were all low in the supernatants during 24 h.In the imiquimod treatment group,the concentrations of IFN-γ were increased significantly,and the concentrations of IL-4 and IL-5 were decreased in the super natants.It worked at 6 h,and achieved the peak at 12 h,lasted over 24 h.In the control group,the expres sions of T-bet and GATA-3 were detected in CD4+T cells during 24 h.In OVA treatment group,the expressions of T-bet were decreased,and that of GATA-3 were increased rapidly in CD4+T cells.In dexam ethasone treatment group,the expressions of T-bet were always low in CD4+T cells,and that ofGATA-3 were no change during 24 h.In imiquimod treatment group,the expressions of T-bet were increased,andthat of GATA-3 were decreased in CD4+T cells.The protein expressions worked at 6 h.and achieved the peak at 12 h,lasted over 24 h.Conclusion The imbalance T cell-specific transcription factors T-bet/GA-TA-3 contributes to both high expression of GATA-3 and low expression of T-bet in CD4+T cells from OVA sensitized mice.Dexamethasone treatment inhibits the expression of T-bet in CD4+T cells and has no func tion in GATA-3.Imiquimod treatment modulates key master switches GATA-3 and T-bet that results in com mitting T helper cell to a TH 1 phenotype and imiquimod may play a key role in the regulation of TH2 cytokine responses in asthma.

In order to obtain enough fusion protein for developing preclinical studies of IFNbeta-HAS, we screened Pichia pastoris transformants expressing high-level protein by immunology method. The yield of IFNbeta-HSA was about 500 mg/L by fed-batch fermentation. The purity of IFNbeta-HSA reached 96% through the steps of ultrafiltration, Blue Sepharose FF, Ni2+-IMAC and DEAE Sepharose FF. Analysis of Western blotting showed that IFNbeta-HSA had the antigenicity of IFNbeta and HSA. The specific activity was about 1.96 x 10(7) IU/mg by standard survival activity test on WISH cells challenged with VSV virus. This study provided a method to produce IFNbeta-HSA.
Fermentation ; Humans ; Interferon-beta ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Serum Albumin ; biosynthesis ; genetics