1.Effects of combination therapy of tirofiban and reteplase on endothelial function,coagulation function and inflammatory factors in elderly patients with acute ST-segment elevation myocardial infarction
Zhuibo YU ; Wanwan CHEN ; Zhongxian XUE
Chinese Journal of Primary Medicine and Pharmacy 2017;24(12):1891-1895
Objective To investigate the effect of the combination therapy of tirofiban and reteplase on endothelial function,coagulation function and plaque inflammation in elderly patients with ST-elevation acute myocardial infarction (STEMI).Methods 100 patients with STEMI were treated with percutaneous transluminal coronary intervention (PCI) from January 2014 to June 2016 in our hospital.39 cases in the control group used conventional oral aspirin,clopidogrel and statins and other treatment.61 cases in the observation group received tirofiban and reteplase on the basis of the control group.The expression of endothelial microparticles (EMP) was detected by flow cytometry (FCM),and the ICAM-1,high sensitivity C-reactive protein (hs-CRP),tumor necrosis factor α(TNF-α) and interleukin-6 (IL-6),endothelin-1(ET-1) were measured by enzyme-linked immunosorbent assay (ELISA).The thrombin time (TT),activated partial thrombin time (APTT),prothrombin time (PT) and other indicators were measured by PUN-2048A coagulation instrument,then statistical analysis was performed.Results The postoperative levels of EMP,ICAM-1 and ET-1 of control group were (693.46±90.72),(768.58±20.46)μg/L and (31.27±8.18)ng/L,which were significantly higher than those in the observation group [(652.36±67.39),(752.37±25.0)μg/L,(28.22±5.05)ng/L],the differences were statistically significant (t=2.41,2.67,2.68,all P<0.05).After operation,the hs-CRP,TNF-α,IL-6 levels in the control group were (4.16±2.35)mg/L,(4.32±2.02)ng/L,(10.59±3.16)ng/mL,which were significantly higher than those in the observation group [(2.22±1.47)mg/L,(2.74±1.52)ng/L,(6.33±2.24)ng/mL],the differences were statistically significant(t=2.65,2.67,3.42,all P<0.05).The postoperative TT,PT,APTT in the observation group were (26.31±3.18)s,(14.34±1.67)s,(27.20±4.12)s,which were significantly longer than those in the control group [(24.03±2.84)s,(12.56±1.43)s,(24.55±3.62)s],the differences were statistically significant(t=2.15,2.31,2.65,all P<0.05).Conclusion Tirofiban combined with reteplase can improve endothelial function,inhibit inflammatory reaction and regulate coagulation function.
2.Construction and expression of growth differentiation factor-5 gene prokaryotic expression plasmid and its cartilage induction activity in vivo
Kangrong LU ; Wanshan WANG ; Xia XUE ; Zhongxian PIAO ; Yingjie PIAO
Chinese Journal of Tissue Engineering Research 2009;13(50):9842-9845
BACKGROUND: Growth differentiation factor-5 (GDF-5) plays an important role in the development and formation of cartilage, extremities, and joints, which is a widely used joint development marker.OBJECTIVE: To express mature peptide of human GDF-5 in E. coil by the way of genetic engineering, and to explore the inductive activity of recombinant protein in vivo.DESIGN, TIME AND SETTING: The observation experiment based on gene was performed at the Analysis and Testing Center of Southern Medical University from January to June 2006.MATERIALS: Human fetus cartilage tissue was harvested from Department of Gynaecology and Obstetrics, and the consent was obtained from the family. Ten KM mice were purchased from experimental animal center of Southern Medical University, half male and half female, weighing 18-22 g, aged 6-8 weeks.METHODS: The hGDF-5 gene encoding mature peptide was gained by RT-PCR from the total RNA which was extracted from fetus cartilage tissues, and was inserted into the pET22b(+) vector to construct recombinant prokaryotic expression plasmid pET22b(+)-GDF5, which was transformed into E. coil BL-21 to be expressed after IPTG induction. Proteins of interest were purified with sepharose chelated with nickel ions (Ni2+) and then implanted in mouse hindlimb muscle to evaluate the biological activities by routine hematoxylin-eosin staining.MAIN OUTCOME MEASURES: The expression, sequencing of target gene was observed by agarose gel electrophoresis, and the protein expression was detected by SDS-PAGE electrophoresis, meanwhile, the GDF5-inducing activity was evaluate by histological observation.RESULTS: RT-PCR product was about 350 bp in length, which was confirmed by double enzyme digestion of the recombinant plasmid, sequencing result was in agreement with the reported hGDF-5 sequence in Genbank. SDS-PAGE analysis showed a conspicuous band representing a new foreign protein with relative molecular mass of approximately 14 KD after induced expressioin. Cartilage tissues were formed in the mouse muscle where the purified proteins were implanted. CONCLUSION: The integral human GDF-5 mature peptide gene was cloned successfully from human fetus cartilage tissue and a high-yield expression was achieved in E. coli, the pudfied protein has chondrogenic activities in vivo.