Abstract:Aim It was presumed that Chronic orally organophosphorus insecticide may consume PON1 and result in development of atherosclerosis.The aim of the present study was to explore whether chronic orally organophosphorus insecticide trichlorfon has a similar or aggravating atherogenic effect of hyperlipidic diet and analyze the possible mechanisms.Methods 32 healthy male New Zealand rabbits were divided randomly into 4 groups with 8 rabbits in each group.They were fed a standard normal chow,normal chow plus trichlorfon 18 mg/(kg·d),the hyperlipidic(50 g/d)chow which containing 1% cholesterol and 7.5% yolk powder and 8% lard and trichlorfon plus hyperlipidic chow respectively.After 70 days,rabbites were sacrificed,the function,parameters of both biochemistry and pathohistology in isolated vesseles and blood were analyzed.Results The treatment of trichlorfon and hyperlipidic diet resulted in a significant inhibition of the ACh-induced endothelium-dependent relaxation response,but did not in sodium nitroprusside induced endothelium-independent relaxation in isolated aortic rings from rabbits,a significant decrease of activity of both paraoxonase and superoxide dismutase and a significant increase of concentration of malondialdehyde in sera.An activity of cholinesterase was markedly decreased in the alone trichlorfon treated group,but was significantly increased in hyperlipidic diet group.Hyperlipidic diet resulted in pathologic changes of atherosclerosis examined by light microscopy including that the vascular intima become more rough and thicker than normal,a lot of lipid foam cells migrated to regions under intima interval and smooth muscle cells were arranged in more disorder than normal group,trichlorfon alone did not,but aggravated significantly those changes induced by hyperlipidic diet.Conclusion A long time orally trichlorfon could result in injuries of vascular endothelial function and accelerate atherogenic effects of hyperlipidic diet.The mechanism may be related to the decrease of PON1 activity in sera and oxidative stress reaction induced by trichlorfon.

Abstract:Aim To study the function of amlodipine on cellular cholesterol accumulation and cyclophilin a expression in foam cells derived from smooth muscle cell induced by oxidized low density lipoprotein(ox-LDL) to explore the new antiatherosclerotic role of Amlodipine.Methods The experiment groups included 6 groups:control group,ox-LDL group,different concentration(0.1,1.0,10.0 μmol/L)of amlodipine pretreated for 1 h and treated with 80 mg/L ox-LDL for 72 h group and amlodipine treated alone without ox-LDL group.RT-PCR and Western-blot analysis were used to detect the mRNA and protein expression of cyclophilin A.Oil red O dye experiment showed the lipid droplets in the cells of each group.High Performance liquid chromatography(HPLC) analysis was peformed to measure the content of cellular total cholesterol and cholesterol ester.Results RT-PCR and Western-blot analysis showed that with ox-LDL and amlodipine treatment,cyclophilin A expression were greatly increased in a concentration-dependent manner.Oil Red O dye showed that with the treatment of amlodipine,the lipid droplets were greatly decreased in a concentration-dependent manner vs ox-LDL group.The content of cellular total cholesterol and cholesterol ester were both significantly increased with ox-LDL treatment but decreased after amlodipine treatment.Conclusion Amlodipine can up-regulate the expression of cyclophilin A and relieve the cellular cholesterol accumulation in foam cells derived from smooth muscle cells induced by ox-LDL.

Abstract:Aim To investigate the effects of interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) on endothelial lipase(EL) in rat aortic smooth muscle cell(ASMC).Methods The rat ASMC were obtained from thoracic aortas and cultured by tissue explant method.Passage 3～5 cells were used in experiment,and were randomly divided into 3 groups:GroupⅠ:treated without intervention as the control;GroupⅡ:the rat ASMC were treated in serum-free culture medium containing IL-1β(1 to 30 μg/L) for 24 hours;GroupⅢ:TNF-α(5 to 50 μg/L) for 24 hours.The levels of EL were tested by the method of enzyme linked immunosorbent assay(ELISA) and reverse transcription polymerase chain reaction(RT-PCR).Results Smooth muscle cells were treated with increasing concentrations of IL-1β,ranging from 1 μg/L to 30 μg/L.The relative increases in EL protein and mRNA were greater at the highest concentrations(30 μg/L) of IL-1β.There was a significant difference between the intervention group and the control group(P<0.05).There were significant differences among the intervention groups(P<0.05).The relative increases in EL protein and mRNA were greater at the highest concentrations(50 μg/L) of TNF-α.There was a significant difference between the intervention group and the control group(P<0.05).There were significant differences among the intervention groups(P<0.05).Conclusion Smooth muscle cells were treated with increasing concentrations of IL-1β and TNF-α,then secreted protein and EL mRNA were noted.

Abstract:Aim To investigate the effect of rosiglitazone(RSG) on glucose-induced adhesion of human endothelial cells.Methods Endothelial cells were incubated with different concentrations of glucose,and then treated with different concentrations of rosiglitazone.Vascular cell adhesion molecule-1(VCAM-1) protein and mRNA expression was detected by immunocytochemical method and RT-PCR.Endothelial-monocyte adhesion assays were carried out in treated cells.Results VCAM-1 mRNA and protein expression were increased significantly with glucose concentration increased,which was obvious at 16.7 mmol/L glucose group(P<0.05).Compared with RSG untreated group,relative VCAM-1 mRNA and protein expression decreased to varying degrees in the RSG treated group(P<0.05).Furthermore,endothelial-monocyte adhesion increased with increment of glucose concentration(P<0.05) and decreased after treatment of high dose RSG(P<0.05).Conclusions RSG may appear to have direct anti-diabetes-associated-atherosclerosis which is at least partly due to effects on VCAM-1 expression induced by glucose,which leads to decreased endothelial-monocyte adhesion and macrophage infiltration.

Abstract:Aim To study the basic biological characteristics of adult adipose-derived mesenchymal stem cell(ADMSC) and the potential of differentiating into smooth muscle cells by chemical induction.Methods ADMSC were isolated and cultured from adult greater omentum adipose tissue in vitro.The morphological characteristics of cells were observed by the inverted microscope.The cell cycle and expression of CD29,CD31,CD34 and CD44 were detected by flow cytometry.The fifth passage cells were induced with 5 μg/L transforming growth factor-β(TGF-β) to differentiate into smooth muscle cells.Then the immunohistochemistry method was used to identify smooth muscle cells.Results ①The isolated and cultured ADMSC presented even fusiform shape and stably passaged,during which ADMSC kept the same shape.② ADMSC expressed CD29 and CD44 but not CD34 or CD31 by flow cytometry.③ The cell cycle studies revealed that there were 90.14% of the cells in G0/G1,3.77% in S and 6.09% in G2/M.④ After 10 days induction by TGF-β,the cells assumed a spindle shape and a “hill-and-valley” growth pattern.⑤ The induced cells expressed characteristic α-smooth muscle actin(α-SMA) by immunohistochemistry.Conclusion Adult adipose tissues contain plenty of productive mesenchymal stem cells that can be easily isolated,cultured and differentiated into smooth muscle cells in vitro.

Abstract:Aim To investigate the protective effect of proanthocyanidins(PC) on the injury of vascular endothelial cell(VEC) induced by hydrogen peroxide and its corresponding mechanisms.Methods The endothelial cell strain from human umbilical vein was cultured and a model of VEC injured by hydrogen peroxide(H2O2) was established.This study was conducted as follows:normal control group,H2O2-injury group(250 μmol/L),PC protective groups(25,50,100 mg/L).Cell viability was tested by using MTT.The levels of malondialdehyde(MDA),superoxide dismutase(SOD) and lactate dehydrogense(LDH) activity were measured with corresponding kit.H2O2-induced apoptosis was detected using staining with Hoechst 33258.Protein levels were examined by Western-blot.Results PC could inhibit the hydrogen peroxide induced VEC reduction,suppress LDH activity,reduce the MDA production,and enthance SOD activity under hydrogen peroxide.Typical apoptotic cells were detected using staining with Hoechst 3325.The bcl-2 protein expression increased.Conclusions These results demonstrated that PC can protect the VECs from H2O2-induced injury,and the possible mechanism may refer to its effect of anti-apoptosis,anti-lipid peroxidation and SOD activity enhancing.

Abstract:Aim Two-dimensional difference gel electrophoresis(2-D DIGE) and mass spectrum were used to analyse the differential proteins between RAW264.7 cells and foam cells.Methods RAW264.7 cells cultured in vitro was transformed to foam cells by treating with ox-LDL.The total proteins of the RAW264.7 cells and foam cells were extracted and labeled with Cy3 and Cy5.Each Cy3-labeled sample and Cy5-labeled sample were mixed on the same 2-D gel along with a Cy2-labeled mixture of all samples as an internal standard and run on the same gel.All images were analyzed by DeCyder software DIGE technology with internal standard.12 unique proteins were selected,and in which 10 proteins were successfully identified.Results The expression of Stress protein 70,Disulfide bond isomerase and Cytoplasm actin were decreased in foam cells,then the expression of Glucose regulated protein,Eno1 protein,Peroxiredoxin 4,Stathmin 1 and BID protein were increased.Conclusions The proteomics maps of RAW264.7 cells and foam cells were established,and some different proteins were selected and identified.Molecular mechanisms about the formation of foam cells was explored from proteomics-level,and provided new routes and new targets for the intervention study of foam cells transformation to atheromatous plaque.

Abstract:Aim To investigate circulating levels of endothelial microparticle(EMP) in the plasma of patients with acute coronary syndromes(ACS),and to explore correlations between circulating EMP count and conventional risk factors of cardiovascular disease(CVD).Methods A case-controlled study was conducted in 56 patients with ACS and 26 control subjects without coronary heart disease.Circulating levels of CD31bright/Annexin V+ EMP were measured by flow cytometric analysis.Results Plasma CD31bright/Annexin V+ EMP levels were significantly higher in patients with ACS than those in control subjects(P<0.01).However there was no significant difference in plasma CD31bright/Annexin V+ EMP counts between patients with AMI and those with UAP.Among all subjects,patients with type 2 diabetics had increased plasma CD31bright/Annexin V+ EMP as compared with patients without diabetics(P<0.05).On multivariate analysis,serum triglyceride level was the major independent factor related to plasma CD31bright/Annexin V+ EMP count(r=0.28,P<0.05).In patients with ACS,linear regression analysis showed a strong positive correlation between serum high sensitivity C-reactive protein level and plasma CD31bright/Annexin V+ EMP count(r=0.31,P<0.05).Conclusions Plasma CD31bright/Annexin V+ EMP increased in patients with ACS or diabetics,and plasma CD31bright/Annexin V+ EMP count was correlated with conventional risk factors for CVD.These findings suggest that the more critical endothelial damage is present in ACS patients and type 2 diabetics patients,increased level of plasma CD31bright/Annexin V+ EMP is relative with high concentration of plasma hs-CRP and triglyceride.

Abstract:Aim To explore the relationship between plasma visfatin level and coronary artery atherosclerosis(As) in patients with coronary artery diseases(CAD).Methods 62 causes were undergone coronary angiography and divided into control group of 13 cases and As group of 49 cases.The severity of coronary stenosis was determined by Gensini score and based on the results,the As group were classified into two sub-groups:mild and severe groups.The plasma levels of visfatin were measured by ELISA kits.Results Compared with control group,plasma level of visfatin was significantly elevated in As group.In As groups,patients with severe As have higher plasma level of visfatin than those in patients with mild As.Moreover,plasma level of visfatin was positively correlated with Gensini(r=0.359,P<0.01).Conclusions The plasma level of visfatin is closely correlated with the severity of coronary artery in CAD patients,suggesting that visfatin may be involved in the process of As.

Abstract:Aim To investigate the correlation between the protein expressions of matrix metalloproteinase-9(MMP-9),tissue inhibitor of metalloproteinase-1(TIMP-1) and phospho-c-Jun in peripheral blood mononuclear cells of patients with coronary heart disease and the characteristics of atherosclerosis plaque combined with the coronary angiography.Methods 55 patients who had coronary heart disease and underwent an immediate or selective coronary angiography(CAG) were selected and divided into 3 groups:acute coronary syndrome(ACS) group(n=24),stable angina(SA) group(n=16) and conrol group(n=15).MMP-9,TIMP-1 and phospho-c-Jun expressions in peripheral blood mononuclear cells were measured by Western-Blot.Coronary angiograms were scored according to Gensini integration and analysis of the relationship between MMP-9,TIMP-1,phospho-c-Jun,MMP-9/TIMP-1 and Gensini integration by linear correlation.Results ①MMP-9,TIMP-1,phospho-c-Jun and MMP-9/TIMP-1 were significantly increased in ACS group than those in SA group and control group(all P<0.001).②MMP-9,phospho-c-Jun and MMP-9/TIMP-1 were positively correlated with Gensini integration(separately r=0.433,r= 0.476,r=0.457,all P<0.01),but there was no correlation between TIMP-1 and Gensini integration.③MMP-9 was greatly correlated with phospho-c-Jun in patients with coronary heart disease(r=0.839,P<0.001).Conclusions ①MMP-9 and MMP-9/TIMP-1 were significantly increased but imbalanced in the patients with ACS,which suggested a prediction of more complex damage and more vulnerable plaque in coronary artery.②The activation of c-Jun may contribute to MMP-9 expression during the pathogenesis of atherosclerosis.

Abstract:Aim To determine the platelet surface expression of CD62P and CD40L in type 2 diabetes mellitus patients(T2DM)with coronary heart disease(CHD)and to explore the relationship between them and serum high-sensitivity C reactive protein(hs-CRP)level.Methods The platelet surface expression of CD62P and CD40L were measured by flow cytometry at fasting state in 97 consecutive patients,including 34 CHD patients,33 T2DM patients,and 30 T2DM with CHD patients.30 subjects were served as a control group.The correlation between platelet CD62P and CD40L and serum level of hs-CRP were investigated.Results Platelet CD62P and CD40L levels and serum level of hs-CRP in the T2DM with CHD patients were significantly higher than those in the control group,CHD patients and T2DM patients(P<0.01).The levels of platelet CD62P and CD40L were positively correlated with serum level of hs-CRP(r=0.343 and 0.495,both P<0.05).Conclusions Platelet surface expression CD62P and CD40L levels,which are significantly higher in the T2DM with CHD patients and were positively correlated with serum level of hs-CRP,may be useful markers for predicting the presence of coronary artery disease in T2DM patients.

Abstract:Aim To delineate the plaque morphologic characteristics among stable angina pectoris(SAP) and acute coronary syndrome(ACS) by intravascular ultrasound(IVUS).Method 51 SAP patients with 63 vessels were recruited and 32 ACS patients with 46 vessels were checked by IVUS.Plaques were divided into soft,fibrous,calcified.The IVUS results of plaque compositions detection were compared between SAP and ACS.Results The ration of soft plaque,extent of remodeling index,external elastic membrane area(EEMA),plaque burden(PB),plaque rupture increased in ACS group.Thrombosis detection rate were not statisticly different between two groups.Conclusion The criteria of soft plaque located in the proximal part of main coronary branches or any detected soft plaque with positive remodeling index and minimal lumen area≤4 mm2 predicted the vulnerable plaque in ACS and deserved the aggressive treatment.

Abstract:Aim To explore the changes of peroxisome proliferator-activated receptor-γ(PPAR-γ),nuclear factor-κB(NF-κB),metalloproteinase-9(MMP-9) and metalloproteinase inhibitor-1(TIMP-1) expressed by peripheral blood monocyte-derived macrophages(MDM) in patients with acute coronary syndrome(ACS) and the relations among them.Methods 48 patients with ACS and 28 cases with stable angina pectoris(SAP) were admitted.A 20 mL fasting blood sample was obtained using a blood cell-separator tube.Peripheral blood mononuclear cells were isolated,and macrophage colony-stimulating factor was added to train them for MDM.After CD40 ligand stimulating,the concentrations of MMP-9 and TIMP-1 in the supernate of MDM and the expression strength of PPAR-γ,MMP-9,TIMP-1 mRNA and NF-κB subunit P65(NF-κB P65) by MDM were determined.Results In ACS group,the expression strength of PPAR-γ mRNA was lower than the SAP group(0.24±0.04 vs 0.39±0.06;P<0.001),while the expression strength of NF-κB P65 was higher than the SAP group(0.42±0.06 vs 0.27±0.02;P<0.001),both the concentration of MMP-9 and TIMP-1 in the supernate of MDM were significantly higher than the SAP group(231.11±51.47 μg/L vs 126.06±13.2 μg/L,P<0.001;139.80±31.54 μg/L vs 112.250 vs 12.68 μg/L,P<0.05),the expression strength of mRNA of MMP-9 and TIMP-1 were higher than the SA group(0.67±0.11 vs 0.24±0.05,P<0.001;0.42±0.09 vs 0.33±0.06,P<0.05).The expression strength of PPAR-γmRNA were negatively correlated to NF-κB P65 and the expression strength of MMP-9(P<0.001),but not correlated to the expression strength of TIMP-1(P>0.05);the expression strength of NF-κB P65 were positively correlated to the expression strength of MMP-9 and TIMP-1(P<0.001).Conclusion For MDM of patients with ACS,the expression strength of PPAR-γ was down-regulated,while the expression strength of NF-κB and MMP-9 was up-regulated.The PPAR-γ may regulate MMP-9 expression before transcribed by regulating the activity of NF-κB.TIMP-1 expression may not be regulated by PPAR-γ.

Abstract:Aim To study some specific high risk subgroups of patients with acute inferior myocardial infarction,such as right ventricular involvement and explore the clinical implication and prognosis influence of right ventricular infarction and the effect of reperfusion therapy.Methods 304 patients of acute inferior myocardial infarction were treated with reperfusion therapy or conservative therapy according to guideline and patients selection.There were 232 patients of inferior myocardial infarction without right ventricular involvement and 72 patients with right ventricular involvement.Datas were recorded including clinical features and complications.Results The incidence of complications significantly increased in patients with right ventricular infarction,which included cardiogenic shock,mechanical complications,complete atrioventricular block,ventricular fibrillation,sustained ventricular tachycardia and reinfarction.The in-hospital mortality was 8.6% in patients without right ventricular involvement and 34.7% in patients with right ventricular involvement.In patients with right ventricular involvement,the in-hospital mortality was 27.8% in reperfusion therapy group and 55.6% with conservative therapy group.Conclusions Right ventricular involvement is a high risk subgroup of acute inferior myocardial infarction,which increases in-hospital morbidity and mortality.Reperfusion therapy can improve the in-hospital outcomes of right ventricular infarction.

Abstract:Aim To investigate the correlation of plasma brain natriuretic,C-terminal portion of pro-vasopressin(Copeptin) and serum high sensitivity C-reactive protein in evaluating the prognosis of patients with acute coronary syndrome.Methods The levels of plasma brain natriuretic,Copeptin and high sensitivity C-reactive protein were measured in 30 patients with acute myocardial infarction(AMI),38 patients with unstable angina pectoris(UAP),and 20 controls.The numbers of impaired coronary arteries,left main coronary stenosis,Gensini scoring of coronary stenosis and the levels of plasma brain natriuretic,Copeptin and high sensitivity C-reactive protein were compared respectively.Results The levels of plasma brain natriuretic,Copeptin and high sensitivity C-reactive protein in group AMI and group UAP were significantly higher than those in the controls(P<0.01),the levels of them in group AMI were significantly higher than those in group UAP.There were obvious positive correlations in the levels of plasma brain natriuretic and Copeptin with impaired coronary arteries,left main coronary stenosis,and Gensini score;however,there were no correlations in the levels of serum high sensitivity C-reactive protein with them.Conclusions Plasma levels of plasma brain natriuretic,Copeptin and high sensitivity C-reactive protein correlates with coronary artery stenosis in patients with acute coronary syndrome.They are important clinical parameters to reflect the severity of myocardial necrosis.

Abstract:Aim To summarize the results of coronary artery bypass grafting(CABG) for the patients with low left ventricular ejection fraction(LVEF).Methods A retrospective study was carried out.63 cases with low LVEF(≤0.40) underwent CABG,45 males and 18 females.Cardiopulmonary bypass(CPB) was applied in CABG.Surgical technique,complications,LVEF,left ventricular end-diastolic diameter(LVEDD),heart function and post-operative treatment were analyzed.Results Early post-operative complication included:Re-open in 1 case(1.6%),LCOS in 5 cases(7.9%),IABP used in 3 cases(4.8%),repeatedly atrial fibrillation in 6 cases(9.5%),complication of CNS in 1 case(1.6%),hepatic dysfunction in 2 cases(3.2%),renal insufficiency in 2 cases(3.2%),respiratory insufficiency in 3 cases(4.8%),pleural effusion in 7 cases(11.1%) and death in 1 case(1.6%).The others had favorable outcomes.There were significant improvement in mean LVEF,LVEDD and NYHA class postoperatively(P<0.01).Conclusions Suitable adaptative disease,excellent surgical technique,advanced myocardial protection,IABP applied early when weanning from pump is difficult and enhancing post-operative treatment can significantly improve the results of CABG for the patients with low LVEF.