Abstract:Atherosclerosis and its associated complications are the primary cause of death in humans.Aging is the main risk factor for atherosclerosis,compared with any other risk factor.After the age of 60 years,age dominates all other risk factors in the risk prediction.But the specific mechanism that contributes to the aging risk remains unknown.For the past few years,the studies on correlations of stem or progenitor cells with disease made great progress.The correla tions of vascular progenitor cells with vascular disease are also paid more and more attention.In 1997,Asahara reported,for the first time,the isolation of putative endothelial progenitor cells(EPC) in the circulating blood.These cells were able to differentiate into mature endothelial cells in vitro and to incorporate into sites of active angiogenesis.This discovery led to a new era of vascular biology and a novel understanding of atherosclerosis and thromboembolic complications.A novel concept for atherosclerosis risk implicates a lack of endothelial progenitor cell(EPC)-dependent arterial repair in the development of atherosclerosis that is secondary to exhaustion of repair-competent EPC.And smooth muscle progenitor cell is one of the sources of smooth muscle cell derived foam cell.This article focuses on the effects of progenitor cells on atherosclerosis,especially on the correlations of endothelial progenitor cell,smooth muscle progenitor cell with atherosclerosis,and the differentiation of vascular progenitor cells.

Abstract:Aim To study the effect of transfection of adenovirus-mediated integrin-linked kinase(ILK) on rat bone marrow mesenchymal stem cell(BMSC) paracrine action.Methods BMSC were isolated and cultured in vitro by adherent culture.The recombinant adenoviral vector was constructed containing both human wild-type ILK cDNA and humanized recombinant green fluorescent protein(hrGFP).Corresponding virus with null content(adeno-null) was used as a control.BMSC were infected by adeno-ILK(MSC-ILK group) or adeno-null(MSC group) in serum-free medium.The infection efficiency of adeno-ILK was tested by GFP expression using flow cytometry,thus the best multiplicity of infection(MOI) was identified.The mRNA was extracted from BMSC and the gene expression of vascular endothelial growth factor(VEGF),fibroblast growth factor-2(FGF-2),and insulin-like growth factor-1(IGF-1) was examined by Real-time PCR.Results The gene expression of VEGF,FGF-2,and IGF-1 in the MSC-ILK group was 8.2±0.4,2.6±0.2 and 2.4±0.2 times(P<0.05 or P<0.01) higher than that in the MSC group respectively.Conclusion BMSC transfected by ILK gene could express higher VEGF,FGF-2,and IGF-1.

Abstract:Aim To investigate the protective effects of Tongxinluo(TXL) on vascular endothelial cells injured by homocysteine and its mechanism.Methods Human umbilical vein endothelial cells(HUVEC) cultivated in vitro were divided into four groups: control group,homocysteine group,TXL group and atorvastatin treatment group.The cell proliferation ability was determined by MTT assay.Real-time reverse transcription polymerase chain reaction(RT-PCR) was performed to detect mRNA expression of superoxidase dismutase(SOD).The content of malondialdehyde(MDA) and SOD activity in HUVEC cellular supernatant were detected.Results Homocysteine significantly inhibited endothelial cell viability(P<0.05).Tongxinluo and atorvastatin significantly improved endothelial cell viability reduced by homocysteine(P<0.05).Homocysteine decreased the expression of SOD mRNA in HUVECs(P<0.01).Tongxinluo and atorvastatin increased the expression of SOD mRNA compared with homocysteine group(P<0.05).Compared with the control group,the activity of SOD in Hcy group decreased significantly(P<0.05) while the content of MDA increased dramatically.However,the activity of SOD in TXL group and atorvastatin group increased significantly compared with Hcy group(P<0.05),at the same time,the concentration of MDA in TXL group and atorvastatin decreased(P<0.05).Conclusions TXL capsule can protect the HUVECS from injury by homocysteine.And it may take an important role in the antioxidant of TXL.

Abstract:Aim Researching the correlation of atherosclerosis related genes and hyperlipidemia,in order to find atherosclerosis early diagnosis target.Methods Middle-aged male crab-eating Macaque(Macaca fascicularis) were fed moderately atherogenic diet(0.22 mg cholesterol/kcal and 40% of calories as saturated fat) for 12 months.According to blood-lipid hyperlipidemia crab-eating Macaques(n=10) were selected,then the expression differences of 113 atherosclerosis related genes were analysed by real-time PCR.Results 66 genes were detected in peripheral blood leucocyte.There were 18 genes upregulation and 21 genes downregulation with elevated blood lipid(P<0.05),besides 27 irregular genes.Conclusions Above result narrowed the scope of genes selection for further research,through further research we may select atherosclerosis early diagnosis target.

Abstract:Aim To study the effect of amlodipine on platelet-derived growth factor-B(PDGF-B) mRNA expression in HUVEC-12 endothelial cells induced by oxidized low density lipoprotein(ox-LDL) to explore the new antiatherosclerotic role of amlodipine.Methods HUVEC-12 endothelial cells were treated with 50 mg/L ox-LDL for 24 hours through the preexperiment.The experiment groups included 5 groups: control group,ox-LDL group,different concentration(0.1,1.0 and 10.0μmol/L) of amlodipine pretreated for 0.5 h and treated with 50 mg/L ox-LDL for 24 h group.RT-PCR was used to detect the mRNA expression of PDGF-B.Results RT-PCR analysis showed that with ox-LDL and amlodipine treatment,the mRNA expression of PDGF-B was greatly decreased in a concentration-dependent manner.Conclusion Ox-LDL can affect the mRNA expression of PDGF-B in the HUVEC-12 endothelial cells.Amlodipine can down-regulate the mRNA expression of PDGF-B.

Abstract:Aim To investigate the effects of advanced glycation end products(AGE-BSA) on oxidative stress and apoptosis in human bone marrow derived Mesenchymal stem cells(hMSC),providing new evidences for a higher survival of donor MSC in the heart of diabetes cardiomyopathy after clinical stem cell transplantation.Methods The adult hMSC were isolated from adult bone marrow and cultured in L-DMEM with 10%FCS,were suspended by trypsin and passaged for subsequent passages.At the third passage,hMSC were identified as the immunophenotype of cell surface CD44,CD105,CD34 on flow cytometry.Then,hMSC were cultured in the absence or presence of AGE-BSA for 24 hours.CCK8 were used to evaluate the proliferation capability,reactivated oxygen species(ROS) were analyzed using the ROS assay kit,the content of malondialdehyde(MDA),the activity of superoxide dismutase(SOD) were measured;the cell apotosis was detected by Annexin V/PI flow cytometry.Results The hMSC exhibited adherent long spindle-shaped cells.The immunophenotype of hMSC showed that both stromal cell-associated markers(CD44) and the stem cell-associated marker(CD105) were positive,which were 97.8% and 98.9%,while blood cell,endothelial cell-associated marker(CD34) was negative,which was only 0.8%.Compared with the control group,co-culturing with 20,50,100,200 mg/L AGE-BSA increased ROS production,the content of MDA in cells and the rate of apoptosis.Simultaneously,AGEBSA decreased antioxidase and proliferation of the cells.Both of them were in a dose dependent manner.Conclusion AGE increases the production of ROS in hMSC,while weakens the production of antioxidases,it increases the oxidative stress,breaks the homeostasis of cells,accordingly,AGE inhibits the proliferation of hMSC and increases the apoptosis of these cells.

Abstract:Aim To observe the binding specificity of aptamers and THP-1 macrophage derived foam cells and atherosclerotic lesions.Methods The FITC-labeled PM1,binding to THP-1 macrophages,vascular smooth cells,endothelial cells and THP-1 macrophage derived foam cells were observed by fluorescence microscopy respectivly.Model of atherosclerosis was formed by high-fat feeding rabbit methods.The binding of aptamers and atherosclerotic lesions were detected by fluorescence microscopy and PCR.Results PM1 didn't bind to THP-1 macrophages,vascular smooth cells and endothelial cells but bind to THP-1 macrophage derived foam cells and atherosclerotic lesions.Conclusions we have obtained PM1 specifically binding to THP-1 macrophage derived foam cells and atherosclerotic lesions.

Abstract:Aim To investigate the effects of lipoxin A4(LXA4) on lipopolysaccharide(LPS)-induced differentiation and mature of RAW264.7 macrophages into dendritic like cells(DC).Methods RAW264.7 murine macrophages were respectively treated with LPS(0 μg/L and 100 μg/L),LPS(100 μg/L) together with LXA4(100 nmol/L and 500 nmol/L).Cells surface markers(CD80,CD86 and MHC-II) were analyzed by FACS,and cell morphology was observed by laser scanning confocal microscope and phase contrast microscope.Results LPS could significantly promote the differentiation of RAW264.7 murine macrophages into DC and the expression of surface molecules(CD80,CD86,MHC-II)(P<0.01),and LXA4 could inhibit the differentiation of LPS-stimulated RAW264.7 murine macrophages and down-regulate the expression of CD86 and MHC-II,but had no significant effect on the expression of CD80.Conclusion LXA4 could negatively regulate the differentiation of RAW264.7 into DC and inhibit the maturation of DC,which may inhibit the pathogenesis and development of atherosclerosis by negatively regulating immuno-inflammatory response.

Abstract:Aim To construct and identify the miRNA eukaryotic expression vectors of(pro)renin receptor((P)RR).Methods miRNA nucletides of(P)RR were chemically synthesized and inserted into pcDNATM6·2-GW/EmGFPmiR vector,which were confirmed by sequencing,then the recombinant miRNA vectors were transfected into A7r5 cell by LipofectamineTM 2000.The mRNA expression of(P)RR was detected by Real-time PCR and Western blot.Results Sequencing suggested that miRNA eukaryotic expression vectors targeting(P)RR possessed correct read frame and nucleotide sequence,and green fluorescene of the transient transfected A7r5 cell lines could be observed under inverted fluorescence microscope.Real-time PCR and Western blot results showed that the sequence of X2-2-3 and X2-3-3 could effectively knockdown the level of mRNA and protein of(P)RR.Conclusions miRNA eukaryotic expression vectors targeting(P)RR were successfully constructed and the effectively interference RNA were identified,which may be used for understanding the effect of(P)RR in the vascular smooth muscle cells.

Abstract:Aim To evaluate monocyte chemoatlractant protein-1 level and the relation of the vascular inflammation factor and coronary vulnerable plaque in the patients with abnormal glycometablism.Methods 86 unstable angina pectoris patients with the coronary vulnerable plaque undergoing coronary angiogram and IVUS were consecuted.36 patients with abnormal glycometablism entered the diabetes group,50 patients with no diabetes in the control group.The serum inflammation factors were tested.Results According to the patients with unstable angina pectoris,the serum monocyte chemoattractant protein-1(MCP-1) in the patients combinated with abnormal glycometablism was more than that of the control groupis.And the multiple regression analysis showed that the MCP-1 was related to the plaque square.Conclusions The unstable angina pectoris patients with abnormal glycometablism have MCP-1,moreover the latter was related to the coronary plaque volume.

Abstract:Aim To explore the relationship of serum level of eosinophilic cation protein(ECP)and classification of coronary artery lesions through measuring their levels.Methods The study enrolls 70 patients with coronary heart disease(CHD),and 30 patients without CHD as controls.All patients were confirmed by angiography.According to the number of diseased vessels,we divided the patients into three groups: 1-vessel disease group,2-vessel disease group and 3-vessel disease group;and then analyzed the relationship between the level of ECP and the number of diseased vessels respectively.Results The serum levels of ECP in CHD group were higher than those in control group(P<0.01).The relationship between the serum levels of ECP and the number of diseased vessels were statistically related among three groups(P<0.01).Multiple linear regression analysis showed that levels of ECP were closely associated with the severity of coronary artery lesions.In multivariate analysis,ECP levels were significantly associated with CHD independent of traditional risk factors.Conclusions The study indicates that the higher serum levels of ECP may be related to the initiation and development of coronary artery lesions.So the higher serum levels of ECP is likely one of the indexes to predict the severity of the coronary artery lesions.

Abstract:Aim To evaluate the incidence and risk factors of atherosclerotic renal artery stenosis(ARAS) undergoing coronary angiography.Methods A total of 279 consecutive Chinese patients with coronary artery desease confirmed by coronary angiography underwent an abdominal aortogram in the same sitting to screen for ARAS.Patient demographics and comorbidities were analyzed for any association with ARAS.Results A total of 80(28.7%) patients was found to have significant ARAS,60(34.3%) of the 175 patients with coronary artery desease who had at least one coronary artery of ≥50% diameter stenosis,20 cases had atherosclerotic renal artery stenosis in 104 patients of normal coronary arteriography.The frequency of ARAS increased with the number of stenotic coronary arteries.By multivariate logistic regression analysis,smoking,pulse pressure(PP),coronary artery stenosis,and serum creatinine were independent predictors of ARAS.Conclusion Abdominal DSA should be routinely performed in patients with coronary artery desease undergoing coronary angiography to identify ARAS,especially in those associated with hypertension and coronary artery disease.

Abstract:Aim To investigate the effect of telmisartan on vascular endothelial function in patients with metabolic syndrome.Methods 76 cases of patients with metabolic syndrome were recruited,and randomized to receive conventional treatment(n=38) or telmisartan(80 mg/d) on the basis of conventional treatment(n=38) for 3 months.Thirty-eight healthy persons were chosen as control group.The plasma level of glucose and lipid metabolism,nitric oxide(NO) and cytokines of each group before and after treament were determined.The blood pressure and endothelial relaxation function were also measured.Results The blood glucose and blood pressure in telmisartan group and conventional group were all decreased after treatment(P<0.01),without intergroup differences(P>0.05).In telmisartan group,the flow-mediated diameter(FMD) and NO were obviously increased(P<0.05 and P<0.01),and the plasma tumor necrosis factor α(TNF-α),interleukin-6(IL-6) and C-reactive protein(CRP) were decreased(P<0.05),with intergroup differences(P<0.05).No obvious changes of FMD,NO and inflammatory factors were found in conventional group.Conclusion Telmisartan can improve vascular endothelial function in patients with metabolic syndrome by raising concentration of NO and inhibiting inflammatory reaction with decreased level of cytokines.

Abstract:Aim To investigate the level of matrix metalloproteinase-3 of peripheral blood from acute coronary syndrome patients and to analyize the relationship among matrix metalloproteinase-3,myocardium enzymes,troponins,blood-fat,and severity of coronary artery disease.Methods To contrast matrix metalloproteinase-3 of peripheral blood from 77 acute coronary syndrome patients of our hospital and normal controls,and to analyze relationship among matrix metalloproteinase-3 of peripheral blood,myocardium enzymes,troponins,blood-fat,and severity of coronary artery disease.Serum level of matrix metalloproteinase-3 was measured by enzyme linked immunosorbent assay.Results Compared with stable angina pectoris group and control group,the level of matrix metalloproteinase-3 in acute myocardial infarction group had statistically significant difference(P<0.01).Compared with stable angina pectoris group and control group,statistical significance of difference was seen in unstable angina pectoris group(P<0.01).There is no significant difference between acute myocardial infarction and unstable angina pectoris group,as well as stable angina pectoris and control group.Matrix metalloproeinase-3 and troponin I,creatine kinase,MB isoenzymes of creatine kinase had positive correlation,There were no correlation between matrix metalloproteinase-3,triglyeride,low density lipoprotein cholesterol.Matrix metalloproteinase-3 level and coronary artery radiography pathological change severity were performed by Pearson correlation: there were no correlation(r=0.293;P=0.053).There were significant difference of matrix metalloproteinase-3 in 10 patients with acute myocardial infarction before and after percutem coronary artery interventional therapy(P<0.01).Conclusion The level of matrix metalloproteinase-3 of peripheral blood in patients with acute coronary syndrome was increased.The level of matrix metalloproteinase-3 and coronary artery radiography severity had no correlation.Matrix metalloproteinase-3 and troponin I,creatine kinase,MB isoenzymes of creatine kinase had positive correlation in patients with acute myocardial infarction.

Abstract:Aim To establish the method of detecting functional lipoprotein(a),and evaluate it,then compare its effect with creatine kinase isozyme at the diagnosis and detection of the acute myocardial infarction.Methods The level of lipoprotein(a) and lipoprotein(a) combining to polyphosphate lysine at preoperative and postoperative 4 h,8 h,12 h,24 h,48 h and 72 h,4～9 days and 10～12 days points were detected respectively in acute myocardial infarction and unstable angina patients by enzyme-linked immunosorbent assay and immune turbidimetric assay.Results The change of lipoprotein(a) level was lower(P>0.05);However,the change of lipoprotein(a) combining to polyphosphate lysine was larger in acute myocardial infarction(P<0.05);The precision and accuracy of the method is better(>90%);The diagnostic value was more superior than creatine kinase isozyme.Conclusion The level of lipoprotein(a) detected by traditional methods fail to associate with the cardiovascular disease;The method that detecting the level of lipoprotein(a) combining to polyphosphate lysine can diagnose and monitor cardiovascular disease,and the method can be used to research the pathological mechanism of lipoprotein(a).

Abstract:The small amount of free fatty acids in serum is not bound to albumin,which is frequently referred to as "the unbound free fatty acids".Unbound free fatty acids and their intracellular binding protein,heart-type fatty acid-binding protein,have been suggested to have clinical utility as indicators of cardiac ischemia and necrosis,respectively.They may have a potential role in identifying patients with cardiac ischemia,predicting recurrent cardiac events in acute coronary syndromes and in congestive heart failure patients.

Abstract:Although arterial conduits are widely used and have improved the long-term results of coronary artery bypass grafting,vein grafts remain important additional conduits in coronary surgery.Newer studies show a saphenous vein graft patency of 60% or more at 10 years postoperatively.Vein graft occlusion occurs as a result of neointimal hyperplasia,which takes place in response to hemodynamic changes,vessel wall injury and thrombosis,and is characterized by the migration and proliferation of vascular smooth muscle cells.Intimal hyperplasia is further complicated by the concomitant development of atherosclerosis.In this review we will summarize the pathogenesis of coronary vein graft disease.

Abstract:Recent studies indicated that oxidative stress played important role in the pathogenesis of human hypertension.We reviewed the production of reactive oxygen species,biomarkers,and mechanisms involved in vascular damage in hypertension,and antioxidative treatment in hypertension.