1.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.
2.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 .
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.
4.Effects of abstragulus mongholicus bung on apoptosis of neurocytes and the expression of relevant gene in the brain of aging mice
Xiaodong WEI ; Zhao WANG ; Hongbo SHAN ; Pengxia ZHANG ; Qin OU
Chinese Journal of Tissue Engineering Research 2006;10(7):151-153
BACKGROUND: The death of aging cells is virtually apoptosis. To a certain extent, it can be interpreted as a series of results of gere activities.Therefore, the inhibition of oncogene's expression can lengthen the life span of cells and delay aging of brain tissues.OBJECTIVE: To study the effect of abstragulus mongholicus bung (AMB) on apoptosis of nerve cells and the expression of relevant gene in aging mice brain.DESIGN: Completely randomized design and controlled experiment.SETTING: Department of Biochemistry, School of Basic Medical Sciences of Jiamusi University.MATERIALS: The experiment was conducted at the Experimental Animal Center and Biochemical Laboratory of Jiamusi University from December 2003 to May 2004. Totally 24 healthy Kunming mice were recruited in this study. There were 8 two-month-old mice (young group) and 16 twelve-month-old mice. All the 16 mice were randomized into abstragulus mongholicus bung group and old control group with 8 in each group.METHODS:① AMB group:Mice in AMB group received gastric gavage was provided by the Department of Traditional Chinese Medicine of the First Hospital Affiliated to Jiamusi University, and evaluated by Jiamusi Drug Inspection Bureau. Water decoction was prepared with 2 kg/L raw materials. Mice in old control group and young group were filled with lukewarm boiled water.② All the animals were treated as above for 30 consecutive days before put to death. Their brains were taken out immediately and the middle parts of the brains were removed to fix with neutral formaldehyde. The remaining brain tissues were made into mitochondria suspension. Content of manganese superoxide dismutase (Mn-SOD) and malondialdehyde (MDA) was determined with xanthosine oxidase method and TBA chemical colorimetry. Apoptotic cells (cells with yellow nuclei were positive ones) were assayed with in situ end-labeling (ISEL) and expression of bcl-2 gene was assayed with immunohistochemical method. The cells stained brown were positive ones. A total of 400 cells were counted under the 400× microscope. We graded the samples according to the percentage of the positive cells: the number of positive cells < 5% -; 5%-10% +; 11%-50% ++; > 51% ().③ Grade and quantitative data were compared with rank sum test and t-test.MAIN OUTCOME MEASURES: Effects of AMB on the rate of neu-ronal apoptosis, the activity of Mn-SOD, the concentration of MDA in mitochondria, and the intensity of the expression of bcl-2 gene.RESULTS: Totally 24 mice entered the final analysis.① Content of MnSOD was higher in young group and AMB group than in old control group (P<0.05).② Concentration of MDA and apoptotic rate in young group and AMB group were lower than those in old control group (P < 0.01).③Expression of bcl-2 gene was significantly different in young group and AMB group from that in old control group (P < 0.01).CONCLUSION: AMB is found to be able to obviously inhibit neuronal apoptosis in aging mice brain by affecting the activity of Mn-SOD, the concentration of MDA and the expression of bcl-2.
7.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.
8.Expression of hepatocyte growth factor and c-met stimulated by high glucose in human kidney fibroblast and its significance
Shan MOU ; Qingyi ZHANG ; Hanfang ZHAO ; Jufang TONG
Chinese Journal of Endocrinology and Metabolism 1986;0(04):-
Objective To observe high glucose induced expression of hepatocyte growth factor (HGF) and c met in human kidney fibroblast. Methods The effects of glucose concentrations on expression of HGF, c met and plasminogen activator inhibitor (PAI) 1 in cultured human kidney fibroblasts were observed by RT PCR. In the same system, the effect of exogenous HGF on the expression of PAI 1 was investigated. Results Human kidney fibroblasts cultured in high glucose concentration (25 mmol/L) showed higher HGF and c met expressions in the early stage and then manifested a gradient decrease of HGF and c met expressions, but PAI 1 expression was gradiently increased. Exogenous HGF resulted in inhibiting PAI 1 expression. Conclusion HGF is a potential anti fibrogenic factor and activates matrix degradation pathways in diabetic kidney by reducing PAI 1 expression.
10.Molecular mechanisms for arecoline against atherosclerosis
Limei SHAN ; Jinchao ZHANG ; Yanling ZHAO ; Wenyu CUI ; Hai WANG
Chinese Pharmacological Bulletin 1987;0(02):-
AIM To investigate the anti-atherosclerosis mechanisms of arecoline. METHODS Fifty-eight male rats were assigned to control, model, arecoline 1 mg?kg -1 and arecoline 5 mg? [FQ(12。46,X-WZ]-kg -1 groups randomly. The rats in model and arecoline-supplied groups were fed with hypercholesterol diet. The following variables were measured: HE staining of rat aorta; serum level of TC, LDL-CHO, HDL-CHO, IL-8, ET-1 and NO; expression of eNOS protein on rat aorta; expression of MCP-1, ICAM-1, CXCR-2 and eNOS mRNA on rat aorta. RESULTS Arecoline increased plasma level of NO and the expression of eNOS protein and mRNA, decreased plasma level of IL-8 and the expression of ICAM-1, MCP-1 and CXCR-2 mRNA on rat aorta. CONCLUSION The anti-atherogenic effects of arecoline seems to be closely involved in increasing plasma level of NO and eNOS protein and mRNA expression, decrease in plasma IL-8 level and down-regulation of the expression of ICAM-1, MCP-1 and CXCR-2 genes.-