1.Imbalance of Th1/Th2 in patients with systemic lupus erythematosus combined with coronary heart disease
Shan ZHAO ; Juan YU ; Hong ZHANG
Chongqing Medicine 2016;(2):216-218,222
Objective To investigate the imbalance of Th1/Th2 cell response in patients with systemic lupus erythematosus (SLE) combined with coronary heart disease .Methods SLE patients ,SLE patients with coronary heart disease and healthy con-trols were enrolled and blood samples were collected .T-bet/GATA-3 ,the transcription factors of Th1/Th2 cells ,were detected by real-time PCR ;the intracellular cytokines IFN-γ and IL-4 in CD4 + T cells were stained by fluorescent antibodies and detected by flow cytometry ;the level of serum IFN-γ and IL-4 were detected by ELISA .Results Comparing with healthy control group ,the ex-pression level of Th1 transcription factor T-bet ,the introcellular secretion of IFN-γ in CD4 + T cells and the serum IFN-γ were all decreased in non-coronary heart disease patients with SLE( P < 0 .05) .Comparing with non-coronary heart disease patients with SLE or healthy control group ,the expression level of Th1 transcription factor T-bet ,the introcellular secretion of IFN-γ in CD4 + T cells and the serum IFN-γ were all increased in patients with SLE combined coronary heart disease(P< 0 .05) ;while the expression level of Th2 transcription factor GATA-3 ,the introcellular secretion of IL-4 in CD4 + T cells and the serum IL-4 were all decreased in patients with SLE combined coronary heart disease(P< 0 .05) .Conclusion There were imbalance towards Th1 cell response in patients with SLE combined coronary heart disease ,which may related to the occurrence and development of disease .
2.HPLC Determination of Berberine Hydrochloride in Xiaoyou Capsules
Chunhui ZHANG ; Zhao YANG ; Xiuming SHAN
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(01):-
Objective To establish a method for the content determination of berberine hydrochloride in Xiaoyou Capsules.Method A PR-HPLC method was adopted with a SHIMADZU RP-ODS C18 column as stationary phase and 0.05 mol/L KH2PO4(adjust pH to 3 with H3PO4)-Acetonitrile(70:30)as mobile phase.The detector wavelength was at 265 nm,column temperature was at 40 ℃and the flow rate was 1.0 mL/min.Results Berberine hydrochloride in Xiaoyou capsules showed a good linearity in the range of 0.010 73~0.214 6 ?g.The average recovery was 99.87 %,RSD was 1.43 %.Conclusion The method is simple,reliable,with a good separation,so it can be used for the quality control of Xiaoyou Capsules.
3.siRNA targeting STAT5 induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721
Zhenjun ZHAO ; Lijie ZHANG ; Gang LU ; Yinjuan ZHANG ; Baoen SHAN
Journal of Third Military Medical University 2002;0(12):-
Objective To investigate the effects of inhibition of STAT5 gene expression by RNA interference technology on apoptosis of human hepatocellular carcinoma cell line SMMC-7721. Methods Three siRNA eukaryotic expression vectors against STAT5 were constructed and transfected with lipofectamineTM 2000 into SMMC-7721 cells. The changes in STAT5 expression were detected by semi-quantitative RT-PCR and Western blot. Cell apoptosis was assayed by flow cytometry (FCM). Results The sequence-specific siRNA could effectively and specifically inhibit STAT5 gene expression at both mRNA and protein levels. The inhibition rates of STAT5 mRNA expression were 70.43%, 43.02%, and 45.07%, respectively. The inhibition rates of STAT5 protein expression were 67.45%, 37.36%, and 41.86%, respectively. At 48 h after transfection, apoptosis rate was 25.61%. Conclusion siRNA against STAT5 can inhibit STAT5 gene expression in SMMC-7721 cells effectively and specifically and induce apoptosis of SMMC-7721 cells. siRNA targeting STAT5 has a great potential value in gene therapy of hepatocellular carcinoma.
5.Inhibitory Effects of Periplocin from Cortex Periplocae on Human Lung Cancer Cell Line QG56
Jing ZHANG ; Guang YANG ; Xuetao ZHAO ; Baoen SHAN ; Jianghui LIU
Tianjin Medical Journal 2014;(3):197-199
Objective To investigate the inhibitory effects of periplocin from cortex periplocae (CPP) on human lung cancer cell line QG56 and to discuss its mechanism. Methods QG56 cells were cultured in vitro. The final concentrations of CPP in control group were 1.25, 2.50, 5.00, 10.00 and 20.00μg/L. QG56 cells were treated with ascending concentration of CPP for 24 h, 48 h and 72 h. The cell proliferation was measured using MTT method. The morphological changes of QG56 cells were observed under inverted microscope. Flow cytometry (FCM) was used to detect the effects of CPP on cell cycle and cell apoptosis. The expression of apoptosis associated gene bax mRNA in QG56 cells was detected by RT-PCR. The expres-sion of bax protein before and after treatment of CPP was examined by SP immunocytochemistry. Results The inhibitory ef-fect of CPP on the proliferation of QG56 cells was increased with the increasing concentrations of CPP and the prolonged du-ration of treatment. The morphological changes were displayed in QG56 exposed to CPP. The results of FCM showed that CPP caused cell cycle arrest at G0/G1 phase. The apoptotic rate of QG56 cells was significantly increased after CPP treatment for 48 h (P<0.05). The expression of bax mRNA was increased in QG56 exposed to CPP. The result of immunocytochemis-try indicated that CPP up-regulated the expression of bax protein. Conclusion CPP showed significant inhibitory effect on human lung cancer cell lines QG56 through inducing cell cycle arrest and apoptosis.
6.Common complication and progress of Percutaneous endoscopic lumbar discectomy
Shan ZHAO ; Hailong ZHANG ; Xin GU ; Xiaofei GUAN ; Shisheng HE
International Journal of Surgery 2014;41(11):780-783
Percutaneous endoscopic lumbar discectomy belongs to minimally invasive spine operation Its superiority includes smalleroperation wound,less bleeding,shorter hospital day,and earlier return to function,conpared with the traditional operation.At the same time,percutaneous endoscopic lumbar discectomy has complications,as the open operation.This paper reviews its common complications,diagnosis,prevention and control.
8.Relationship Between Microalbuminuria and Inflammation Media in Type 2 Diabetes Mellitus
shan, HUANG ; ru-gen, ZHANG ; wen-sui, ZHAO
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(09):-
Objective To study the relationship between microalbuminuria and inflammation media of C-reactive protein(CRP) and interleukin-6(IL-6) in type 2 diabetes mellitus,and to explore the clinical significance.(Methods)Ninety-six patients with type 2 diabetes mellitus were divided into 3 groups according to the urine albumin excretion rate(UAER): normal albuminuria group(NA),microalbuminuria group(MA) and clinical proteinuria group(CP).The serum levels of CRP,IL-6,fasting plasma glucose,creatinine and fasting insulin were measured in all the cases. Results The levels of CRP and IL-6 in MA group were significantly higher than those in NA group(P
9.N-myc downstream regulated gene 2 promotes apoptosis of bladder cancer cells through signal transducer and activator of transcription 3 signaling pathway
Yang ZHAO ; Yaohui MA ; Xiaolei ZHAI ; Nan ZHANG ; Zhongjie SHAN
Chinese Journal of Urology 2017;38(7):537-541
Objective To investigate the effect of N-myc downstream regulated gene 2 (NDRG2) on the apoptosis of bladder cancer cells by regulating the signal transducer and activator of transcription 3(STAT3) signaling pathway.Methods The expression level of NDRG2 in human bladder cancer cell BIU-87 and immortalized cell SV-HUC-1 was detected by Western blot.NDRG2 over expression vector and empty vector control (pcDNA3.1),siRNA-NDRG2,siRNA control were transfected into BIU-87 cells.After transfected 48 h,the expression level of NDRG2,Cleaved caspase 3,STAT3,p-STAT3,JAK2,p-JAK2 were detected by Western blot,the cell proliferation and apoptosis were measured by MTT and flow cytometry.After adding inhibitor AG490 of 45 μmol/L in cultured BIU-87 cells,MTT assay was used to detect cell proliferation and flow cytometry was used to detect the cell apoptosis,Western blot to detect the expression level of Cleaved caspase 3,STAT3,p-STAT3,JAK2,p-JAK2.Results The expression level of NDRG2 in bladder cancer cells was higher than that in bladder epithelial cells.The cell survival rate of pcDNA3.1/NDRG2 group was lower than that of pcDNA3.1 group,the difference was statistically significant (P < 0.01).The cell survival rate of siRNA-NDRG2 group was higher than that of siRNA control group (P< 0.01).The apoptosis rate of pcDNA3.1/NDRG2 group was higher than that of pcDNA3.1 group (P < 0.01).The apoptosis rate of NDRG2 siRNA group was lower than that of siRNA control group (P < 0.01).The level of p-STAT3 and p-JAK2 in pcDNA3.1/NDRG2 group was lower than that in pcDNA3.1 group (P< 0.01).The survival rate and apoptosis rate of BIU-87 cells cultured with AG490 were in agreement with the trend of pcDNA3.1/NDRG2 after transfection.Conclusions NDRG2 could promote the apoptosis of bladder cancer cells,and its mechanism may be related to STAT3 signaling pathway.
10.Two Novel Complex Mutations of CYP21A2 Gene Causing Simple Virilizing 21-hydroxylase Deficiency
Xiaoli WANG ; Nan ZHANG ; Yuyan ZHAO ; Xiaoguang SHI ; Zhongyan SHAN
Journal of China Medical University 2014;(5):396-400,406
Objective To investigate the clinical manifestations,laboratory findings and genetic characteristics of two cases with simple virilizing (SV)21-hydroxylase(21OHD). Methods Clinical manifestations and laboratory data were obtained. The promoter and coding areas of CYP21A2 gene were directly sequenced. In silico analysis were used to predict the function of mutations. Results Two patients presented severe virilism. The laboratory examinations showed that plasma ACTH,aldosterone,androstendione and testosterone were significantly increased compared with normal individuals. VCT scan showed hyperplasia of bilateral adrenal. Direct sequencing of CYP21A2 gene showed two complex mutations-H62L/V69L and I2g/10InsL,which were not reported previously. In silico analysis(Polyphen)showed the novel mutation V69L could probably damage the function of CYP21A2 protein. Conclusion The combined mutations,H62L/V69Land I2g/10InsL,could be associated with SV type 21-OHD phenotype.