1.Influence of oxidized low density lipoprotein on the proliferation of human artery smooth muscle cells in vitro.
Chenhui, QIAO ; Kailun, ZHANG ; Jiahong, XIA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(1):20-3
The effects of oxidized low density lipoprotein (ox-LDL) on the proliferation of cultured human vascular smooth muscle cells (vSMC) were investigated in vitro. By using NaBr density gradient centrifugation, LDL was isolated and purified from human plasma. Ox-LDL was produced from LDL by being incubated with CuSO(4). ox-LDL was then added to the culture medium at different concentrations (35, 60, 85, 110, 135 and 160 microg/mL) for 7 days. The influence of ox-LDL on vSMC proliferation was observed in growth curve, mitosis index, and in situ determination of apoptosis. The data were analyzed with SPSS 10.0 software. The results showed that the ox-LDL produced in vitro had a good purity and optimal oxidative degree, which was similar to the intrinsic ox-LDL in atherosclerotic plaque. ox-LDL at a concentration of 35 microg/mL demonstrated the strongest proliferation inducement, and at a concentration of 135 microg/mL, ox-LDL could inhibit the growth of vSMC. ox-LDL at concentrations of 35 and 50 microg/mL presented powerful mitotic trigger, and with the increase of ox-LDL concentration, the mitotic index of vSMC was decreased gradually. ox-LDL at higher concentrations promoted more apoptotic vSMCs. ox-LDL at lower concentrations triggered proliferation of vSMCs, and at higher concentrations induced apoptosis in vSMCs. ox-LDL played a promotional role in the pathogenesis and development of atherosclerosis by affecting vSMC proliferation and apoptosis.
2.Experimental study of tissue-type plasminogen activator gene to prevent vein grafts stenosis.
Xionggang, JIANG ; Xiaobin, LIU ; Kailun, ZHANG ; Jiahong, XIA ; Daokang, XIANG ; Long, WU ; Cheng, ZHOU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(3):314-6
The effects of in vivo local expression of recombined human tissue-type plasminogen activator (t-PA) gene on the thrombosis and neointima formation of vein grafts were explored. Jugular vein-to-artery bypass grafting was performed on 72 New Zealand white rabbits. The rabbits were divided into 3 groups according to the different processing methods: transfected t-PA gene group (n = 24), vector group (n = 24) and blank control group (n = 24). Samples of vein grafts were harvested at different time points after surgery. The expression of t-PA gene in vein graft was detected by RT-PCR and the synthesis of t-PA protein by Western-Blot assay. The t-PA activity was measured by chromogenic substrate assay. The Cr51 labeled platelets accumulation in vein grafts was counted. The histopathological changes were compared in intima hyperplasia index among the three groups after operation. The results showed that at the 2nd, 5th, 14th and 28th day after operation, RT-PCR and Western-blot confirmed the expression of t-PA mRNA and protein at the site of gene transfer. The t-PA activity detected on the 2nd, 5th, 14th and 28th day in experimental group was 370.63 +/- 59.44, 344.13 +/- 48.47, 252.87 +/- 51.80 and 161.75 +/- 68.94 U/g respectively, and disappeared on the 60th day and undetected in the control groups. The number of platelets accumulated in the vein grafts in gene group, vector group and blank control group was (85.04 +/- 21.58) 10(6), (225.87 +/- 85.13) 10(6) and (211.7 +/- 78.02) 10(6) respectively. The number of platelets accumulated in gene group was significantly fewer than that in the control groups. Morphometric analysis revealed that intimal hyperplasia was markedly reduced in the t-PA gene group as compared with that in the control groups. It was suggested that the local expression of t-PA gene in vein graft significantly inhibited the accumulation of platelets, thrombosis and concomitant intimal hyperplasia, by which stenosis of bypass graft could be prevented effectively.
3.Influence of Oxidized Low Density Lipoprotein on the Proliferation of Human Artery Smooth Muscle Cells in vitro
Chenhui QIAO ; Kailun ZHANG ; Jiahong XIA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(1):20-23
The effects of oxidized low density lipoprotein (ox-LDL) on the proliferation of culturedhuman vascular smooth muscle cells (vSMC) were investigated in vitro. By using NaBr density gradient centrifugation, LDL was isolated and purified from human plasma. Ox-LDL was produced from LDL by being incubated with CuSO4. ox-LDL was then added to the culture medium at different concentrations (35, 60, 85, 110, 135 and 160 μg/mL) for 7 days. The influence of ox-LDL on vSMC proliferation was observed in growth curve, mitosis index, and in situ determination of apoptosis. The data were analyzed with SPSS 10.0 software. The results showed that the ox-LDL produced in vitro had a good purity and optimal oxidative degree, which was similar to the intrinsic ox-LDL in atherosclerotic plaque. ox-LDL at a concentration of 35 μg/mL demonstrated the strongest proliferation inducement, and at a concentration of 135 μg/mL, ox-LDL could inhibit the growth of vSMC. ox-LDL at concentrations of 35 and 50 μg/mL presented powerful mitotic trigger, and with the increase of ox-LDL concentration, the mitotic index of vSMC was decreased gradually. ox-LDL at higher concentrations promoted more apoptotic vSMCs. ox-LDL at lower concentrations triggered proliferation of vSMCs, and at higher concentrations induced apoptosis in vSMCs. ox-LDL played a promotional role in the pathogenesis and development of atherosclerosis by affecting vSMC proliferation and apoptosis.
4.Regulation of Hypoxic Response Elements on the Expression of Vascular Endothelial Growth Factor Gene Transfected to Rat Skeletal Myoblasts under Hypoxic Environment
XU LEI ; XIA JIAHONG ; ZHANG KAILUN ; XIE AINI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2008;28(5):568-571
Summary: The regulation of hypoxic response elements on the expression of vascular endothelial growth factor (VEGF) gene transfected to primary cultured rat skeletal myoblasts under hypoxic environment was investigated, pEGFP-C3-9HRE-CMV-VEGF vector was constructed with molecular biology technique and transfected to primary cultured rat skeletal myoblasts by lipofectamine in vitro. Gene expression of transfected myoblasts was detected by RT-PCR, Western blot and fluorescence microscope under different oxygen concentrations and different hypoxia time. The results showed that in hypoxia group, the VEGF gene bands were seen and with the decrease of oxygen concentrations and prolongation of hypoxia time, the expression of VEGF mRNA was obviously increased.Under hypoxic environment, the expression of VEGF protein in the transfected myoblasts was significantly increased. EGFP was expressed only under hypoxic environment but not under normoxic environment. It was concluded that hypoxia promoter could be constructed with HRE and regulate the expression of VEGF gene under hypoxic and normoxic environment, which could enhance the reliability of gene therapy.
5.Experimental Study of Tissue-type Plasminogen Activator Gene to Prevent Vein Grafts Stenosis
Xionggang JIANG ; Xiaobin LIU ; Kailun ZHANG ; Jiahong XIA ; Daokang XIANG ; Long WU ; Cheng ZHOU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(3):314-316
The effects of in vivo local expression of recombined human tissue-type plasminogen activator (t-PA) gene on the thrombosis and neointima formation of vein grafts were explored. Jugular vein-to-artery bypass grafting was performed on 72 New Zealand white rabbits. The rabbits were divided into 3 groups according to the different processing methods: transfected t-PA gene group (n =24), vector group (n=24) and blank control group (n=24). Samples of vein grafts were harvested at different time points after surgery. The expression of t-PA gene in vein graft was detected by RT-PCR and the synthesis of t-PA protein by Western-Blot assay. The t-PA activity was measured by chromogenic substrate assay. The Cr51 labeled platelets accumulation in vein grafts was counted. The histopathological changes were compared in intima hyperplasia index among the three groups after operation. The results showed that at the 2nd , 5th , 14th and 28th day after operation,RT-PCR and Western-blot confirmed the expression of t-PA mRNA and protein at the site of gene transfer. The t-PA activity detected on the 2nd, 5th, 14th and 28th day in experimental group was 370. 63±59.44, 344. 13±48.47, 252. 87±51.80 and 161. 75±68.94 U/g respectively, and disappeared on the 60th day and undetected in the control groups. The number of platelets accumulated in the vein grafts in gene group, vector group and blank control group was (85.04 ± 21.58) 106,(225. 87±85.13) 106 and (211. 57±78.02) 106 respectively. The number of platelets accumulated vealed that intimal hyperplasia was markedly reduced in the t-PA gene group as compared with that in the control groups. It was suggested that the local expression of t-PA gene in vein graft significantly inhibited the accumulation of platelets, thrombosis and concomitant intimal hyperplasia, by which stenosis of bypass graft could be prevented effectively.
6.Anti-CD25 monoclonal antibody modulates cytokine expression and prolongs allografts survival in rats cardiac transplantation.
Jiahong XIA ; Xionggang JIANG ; Yi HUANG ; Kailun ZHANG ; Shiliang XIAO ; Chenyuan YANG
Chinese Medical Journal 2003;116(3):432-435
OBJECTIVETo investigate the role of anti- interleukin-2 receptor (CD25) monoclonal antibody in the regulation of cytokine mRNA expression of IL-1beta, IL-2, CD25, IL-4, IL-5, IL-6, IL-10, tumour necrosis factor-alpha (TNFalpha), and interferon-gamma (IFNgamma) in cardiac allografts to elucidate its immunological mechanism and role in rats that have undergone cardiac transplantation.
METHODSThese in vivo studies were conducted using a rat MHC mismatch SD to Wistar heterotopic cardiac transplant model. Simulect, an anti-CD25 antibody, was used to prevent allograft rejection. An increase in the rate of allograft survival was observed. Rats were sacrificed on day 1, 3, 5, 7, 9, 11, 14 post-transplantation and hearts were harvested for further study. Cytokine mRNA expression was determined by semiquantitative RT-PCR.
RESULTSIn the control group, cardiac allografts were rejected at 8.3 +/- 1.7 days after transplantation (x +/- s). The rats who received CsA rejected the cardiac allograft at 26.4 +/- 5.7 days post-transplant. Allograft survival of Simulect-treated rats was 29.2 +/- 7.1 days (P < 0.05 vs controls). Rats treated with simulect and CsA had the longest survival of 55.0 +/- 11.6 days (P < 0.001 vs controls). CD25 mRNA expression in the heart tissue samples of treated rats was undetectable or very weak. However, the untreated group, CD25 expression increased, although anti-CD25 decreased this CD25 expression in the heart graft. Furthermore, in untreated allografts, IL-2, TNFalpha and IFN-gamma were strongly expressed, an effect that markedly decreased after simulect treatment. Finally, IL-4, IL-5, IL-6 and IL-10 expression was strong in anti-CD25-treated allografts.
CONCLUSIONSThese results suggest that anti-CD25 antibody treatment may not only neutralize CD25 activity but also play a role in altering cytokine mRNA expression and prolong the survival of allografts.
Animals ; Antibodies, Monoclonal ; therapeutic use ; Cytokines ; genetics ; Graft Survival ; Heart Transplantation ; immunology ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Receptors, Interleukin-2 ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Homologous
7. Characteristics of acute myocardial infarction caused by spontaneous coronary artery dissection in young female patients
Peina MENG ; Qiang WU ; Yong XIA ; Delu YIN ; Wei YOU ; Zhiming WU ; Chen XU ; Kailun CHEN ; Jue GU ; Dujiang XIE ; Fei YE
Chinese Journal of Cardiology 2018;46(7):536-542
Objective:
To investigate the characteristics of acute myocardial infarction caused by spontaneous coronary artery dissection(SCAD) in young female patients.
Methods:
In this casecontrolstudy,127 young(≤55 years) female patients with acute myocardial infarction onset within 1 week in Nanjing first hospital, Xuzhou central hospital, affiliated hospital of Xuzhou medical university, and Lianyungang first people's hospital were enrolled between January 2013 and February 2017,and the clinical data were retrospectively analyzed. According to their clinical manifestations and coronary angiography(CAG) results,the patients were divided into coronary atherosclerosis disease(CAD) group(CAG evidenced atherosclerosis,
8.Cardiac mechanical dyssynchrony in pacing in different ventricular parts in elderly patients using myocardial metabolic imaging
Xiaoyan SUN ; Kailun XIA ; Zhenqi GU ; Yule NAN ; Wei LI ; Shouli LIN ; Jiacheng TONG ; Tong LU
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(6):605-609
Objective To evaluate the effect of pacing in different parts of the ventricle on left ven-tricular mechanical asynchrony using myocardial metabolic imaging.Methods A total of 56 elderly patients undergoing permanent pacemaker implantation in our hospital from January to November 2023 were recruited and randomly divided into left bundle branch pacing(LBBAP)group and right ventricular pacing(RVP)group,with 28 patients in each group.Another 28 elderly patients who did not undergo pacemaker implantation surgery were selected as the control group.Within 1 week after pacemaker implantation,18F fluorodeoxyglucose(18F-FDG)positron emission tomo-graphy(PET)/CT myocardial metabolism imaging was performed to analyze PET myocardial metabolism images and evaluate left ventricular mechanical synchrony.Results The LVEF was significantly higher in the control group than the LBBAP group and RVP group[(67.68±9.61)%vs(62.71±11.33)%vs(57.36±16.07)%,P=0.012],but no such difference was seen between the LBBAP group and the RVAP group(P>0.05).The LBBAP group had obviously lower pat-tern standard deviation(PSD),phase histogram bandwidth(PHBW),entropy,summed motion score(SMS),summed thickening score(STS),extent of abnormal motion(Mot Ext)and thicken-ing extent(Thk Ext)when compared with the RVP group(P<0.01).There were no statistical significant differences in the terms of PSD,PHBW,Entropy,SMS,STS,Mot Ext,and Thk Ext between the LBBAP group and the control group(P>0.05).Conclusion 18F-FDG PET/CT myo-cardial metabolic imaging can be used to evaluate left ventricular mechanical synchrony in pacing different parts of the ventricle,and LBBAP can obtain better left ventricular synchrony parame-ters than RVP,similar to the control group.