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[Aim] To gain the fusion protein purified GST-AEP.[Methods] The protein GST-AEP was expressed in E.Coli-DH5? as a fusion protein induced by IPTG.The protein was a kind of inclusion body.The purifying and refolding to inclusion body were optimized.The purity of GST-AEP was identified by 12% SDS-PAGE and thin-layer scanning analysis.The quantitation of the fusion protein GST-AEP was done with BCA Protein Assay.[Results] Purity of GST-AEP was higher than 90% and concentration was about 0.163?g/?l.[Conclusion] The fusion protein was highly purified and the method of fusion protein purification from the inclusion body was developed,which was the basis for further study on AEP.
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Objective To observe the effect of marrow mesenchymal stem cells(MSCs)transplantation combined with Qidantongmai tablet and simple MSCs transplantation on myocardial cells apoptosis and heart function in acute myocardial ischemia rats.Method Rat MSCs were isolated by density gradient method,and then cultured and amplified to the third passage labeled with BrdU.16 SD rats were divided into two groups randomly,transplantation combined with Qidantongmai tablet group and simple transplantation group.The isochoric of suspension of Qidantongmai tablet(2 g/kg)and NS were administered to SD rats by gastrogavage respectively for 14 days.The left anterior descending coronary artery of rat was ligated to establish animal models of acute myocardial infarction.The MSCs labeled with BrdU were implanted intramyocardially into the infarct area.After 3 hours,the heart function were detected by an echocardiographic system(HP 5500 system),equipped with a 12 MHz probe.After 4 weeks,the heart function were rechecked and MVD were detected by immunofluorescence,myocardial cells apoptosis were detected by TUNNEL.Results Heart function were improved in both groups,and was more significant in MSCs transplantation combined with Qidantongmai tablet group.Apoptotic index were decreased and the MVD of ischemic zone were increased in transplantation combined with Qidantongmai tablet group than simple transplantation group.Conclusion MSCs transplantation combined with Qidantongmai tablet could improve heart function of acute myocardial ischemia rats,decrease apoptosis of myocardial cells,increase the MVD of ischemic zone,the therapeutic effect were more significant than simple MSCs transplantation.
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Objective To investigate the effects of Qi-reinforcing and blood-activating prescription,Qidan Tongmai Tablet(QDTMT) on the IL-10 in the rat model of myocardial ischemia/reperfusion injury.Methods A model of myocardial ischemia/reperfusion injury was reproduced by ligation of left anterior descending branch of the coronary artery.Thirty-six male SD rats were randomly divided into six groups(n=6):sham operation group,model group,Diltiazem Hydrochloride group,QDTMT high-dose group,QDTMT medium-dose group and QDTMT low-dose group.After the left anterior descending branch of coronary artery was occluded for 30 minutes followed by reperfusion for 2 hours,the whole blood was collected via right common carotid artery and the serum level of IL-10 was measured by ELISA.At the same time,HE staining was performed to detect the pathological change in myocardium,and the ultrastructural alterations of rats were observed with transmission electron microscope.Results The contents of IL-10 in serum were increased in QDTMT groups.Electron microscopic examination showed that pathologic changes of myocardiocytes in QDTMT groups were milder than those of the model group.The effects in QDTMT high-dose group were better than that in the low-dose group,and has insignificant difference with Diltiazem Hydrochloride group.Conclusion Qi-reinforcing and blood-activating prescription inhibits the inflammatory reaction by facilitating the expression of endogenous IL-10 and improving myocardial ultrastructural changes after myocardial ischemia/reperfusion in rats.These actions may contribute to its protective effect on the ischemic myocardial cells.
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Objective To investigate the effect of Qidan Tongmai tablet (QDTMT) on apoptosis and nitric oxide synthase (NOS) of ischemia/reperfusion myocardium in rats. Methods Male Sprague-Dawley (SD) rats were divided randomly into 4 groups:Sham, ischemia/reperfusion (control), QDTMT-Ⅰ (1.08 g/kg) treatment and QDTMT-Ⅱ (3.24 g/kg) treatment. Anesthetized and the open-chest animals were subjected to 40 min of myocardium ischemia followed by 4 h-reperfusion. The cardiomyocyte apoptosis was determined by terminal deoxynucleotidyl transfease-mediated dUTP nick-end labeling (TUNEL) method. The activity of NOS in heart tissue was assessed by chemical assay. Results Pretreatment with QDTMT at different doses reduced cardiomyocyte apoptosis indexes significantly compared with the control (P
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Objective To evaluate the clinical effects in t he treatment of femoral peritrochan teric frac-tures by PFN(proximal femoral nail).Methods A retrospective analysis was done in the treatment of femoral peritrochanteric fractures(72cases).38cases were male and 44cases female.Their age ranged 64～96years old with the average age being 78years old.70cases were intertrochanteric f ractures(EvensⅠtype 16,Ⅱtype 31,ⅢA type 14,ⅢB type 6,Ⅳtype 3),and 2cases subtrochanteric fractu res.Results65patients were followed up for over 6months.All fractures healed and all patients could take care of themselves and were satisfied with t he clinical curative effect except one patient who had the lag screw pulled o ut and coxa vara because of serious os-teoporosis.Conclusion PFNis a better way to cure femoral peritrochanteric fractures because of few complications and simple surgical procedure.Bein g a micro-trauma technique,it is worthy of wide application in clinic.[
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Objective To investigate the effect of curcumine on the expression of matrix metalloproteinase-9 (MMP-9) in human umbilical vein endothelial cell (HUVEC). Methods HUVEC were cultured and identified in vitro .After cultured with curcumine at different concentration for 24 hours, the expression of MMP-9 in HUVEC was detected by cytoimmunochemical staining method and ELISA . Results All concentrations of curcumine can restrain the expression of HUVEC after co- culturing with the cells for 24 hours. The inhibitory effect got stronger with the increase of the concentration of curcumine. Conclusion The curcumine can depress the expression of matrix metalloproteinase-9 in HUVEC obviously.
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Objective To investigate the effects of Qidantongmai Tablet (QDTMT, a Chinese patent medicine) on the dynamic changes in plasma brain natriuretic peptide (BNP) after acute myocardial infarction (AMI) in rabbit induced by ligation of coronary artery, and to explore its influences on ventricular remodeling. Methods Thirty-six healthy rabbits were randomly divided into six groups with six animals for each group: normal group, sham operation group, model group (AMI group, reproduced by ligation of left circumflex branch of the coronary artery), control group (diltiazem hydrochloride group), high dose QDTMT group and low dose QDTMT group. The preventive effect of QDTMT on AMI and its impact on BNP and ventricular remodeling were then investigated. Results ① In high and low dose QDTMT groups, the heart rates increased, the systolic pressure and the pressure differentiation in left ventricle increased obviously, which were significantly different with that of the model group (P