Abstract:Aim To study the cardioprotection of cardiotrophin-1(CT-1) and investigate the signaling pathways involved in the protective effect of CT-1.Methods Cardiomyocytes from the hearts of 1-3-day-old neonatal rats were prepared by a modified method.Five groups were included in the study:control group,hypoxia/reoxygenation group,hypoxia/ reoxygenation+CT-1 group,hypoxia/reoxygenation+CT-1+PD98059(ERK inhibitor) group,and hypoxia/reoxygenation+CT-1+DMSO group.The concentration of CT-1 was 10 μg/L.Myocytes survival rate was evaluated by MTS method,apoptosis,mitochondrial permeability transition pore(Δψm) and reactive oxygen species(ROS) were detected by flow cytometer.The expression of Bad mRNA was measured by RT-PCR,phosphorated ERK1/ERK2 protein level was measured by Western blot.Results Cardiomyocyte apoptosis and ROS(19.4%±2.3% vs 2.2%±0.2% and 14.28±1.42 vs 3.54±0.46;P<0.05) and the expression of Bad mRNA increased markedly after hypoxia/reoxygenation,but cardiomyocyte survival rate and the level of Δψm decreased significantly.Phosphorated ERK1/2 protein level decreased significantly.With CT-1 intervention,cardiomyocyte survival rate increased markedly,apoptosis and ROS reduced significantly.The level of Δψm increased.Expression of Bad mRNA downregulated and phosphorated ERK1/2 protein level increased.The effects of CT-1 could be inhibited by PD98059,which confirmed that PD98059 specifically involved blocking the protective effect of CT-1.Conclusions CT-1 can protect cardiac cells against hypoxia/reoxygenationinjury,these effects are dependent upon its ability to activate the extracellular signal regulated kinase ERK1/2 pathway.

Abstract:Aim To investigate the effect and possible mechanism of bone marrow mesenchymal stem cells(BMSC)transplation on reparing injured vessels endothelium after carotid atherosclerosis stenosis angioplasty in rabbits.Methods Carotid atherosclemsis stenosis model of 48 rabbits had been successfully built up and randomly divided into BMSC transplantation group(n=24) and control group(n=24).BMSC were obtained by density gradient centrifugation and adherent cultured.Surface markers of BMSC were detected by flow cytometry,and BMSC pre-labled by DAPI.Balloon injured carotid artery of rabbits,meanwhile,BMSC(107/kg)were infused into injured artery of BMSC transplantation group rabbits by external carotid artery,and control group infused the same amount of PBS solution.The peripheral blood was collected and the vascular endothelial growth factor(VEGF) levels was detected by enzyme linked immunosorbent assay(ELISA) at preoperative and 3,7,14,28 days of BMSC transplantation.7 days after BMSC transplantation,DAPI labeled BMSC were detected by immunofluorescence microscopy.14 days after BMSC transplantation,the immunohistochemical staining was used to analyse platelet-endothelial cell adhesion molecule(CD31)expression in the injured vessels.28 days after BMSC transplantation,the neointimal area,the ratio of the intima/media area and vascular restenosis were analysed in the vascular tissue by hematoxylin and eosin staining.Results The expression of VEGF in BMSC transplantation group were elevated significantly compared with control group at 3,7,14,28 days after BMSC transplantation.7 days after BMSC transplantation,DAPI labeled BMSC were detected in injured vessels intima.14 days after BMSC transplantation,CD31 continues to express in intima of BMSC transplantation group,while the control group did not express CD31.The neointimal area(0.092 ± 0.009 vs 0.189 ± 0.007,P<0.01),the ratio of the intima/media area(0.698 ± 1.570 vs 1.630 ± 0.122,P<0.01) and the luminal stenosis ratio(41.70% ± 3.70% vs 61.28% ± 1.57%,P<0.01) were significantly decreased in BMSC transplantation group compared with control group at 28 days.Conclusion BMSC transplantation can promote repairing of endothelial after atherosclerotic stenosis carotid artery by balloon injury and reduce the restenosis of injured vessels.

Abstract:Aim To clarify the pathological effects and mechanisms of cellular repressor of E1A stimulated gene(CREG) on tumor necrosis factor-α(TNF-α) stimulated vascular endothelial cell(VEC) injury.Methods CREG overexpressed(VC) and knocked-down(VS) human artery VEC were produced and cells were exposed to TNF-α(10 μg/L).Interleukin-6(IL-6) secretion from cells was determined by enzyme immunolinked assay(ELISA).Filamentous actin(F-actin) stress fibre were detected by Rhodamin-Phalloidin staining.Endothelial permeability was detected by measuring the flux of biotin labeled albumin across the EC monolayers.Nuclear factor-κB(NF-κB) expressions and translocalization were examined by Western blot analysis and immunofluorescence.Results After TNF-α stimulation,ELISA showed that IL-6 expression and secretion in VS cells was markedly increased as well as F-actin cytoskeleton rearrangement.Meanwhile,the permeability of VS cells was detected enhanced obviously.Conversely,overexpression of CREG inhibited the secretion of IL-6,F-actin stress fibre formation and hyperpermeability induced by TNF-α in VC cells. Moreover,immunoflurosence and Western blot showed that NF-κB transiently translocated into the nuclei of VC cells,followed by quick exportation into the cytoplasm.Corresponding changes in the pattern of its expression was also observed.However,the expression of NF-κB in CREG knocked-down VS cells was more sustainably elevated and retained in the nuclei.Conclusions CREG can inhibit NF-κB expression,combat TNF-α-induced inflammatory responses and the hyperpermeability of VEC,and thereby antagonize pathological cellular injury.

Abstract:Aim To explore the influence of Guizhi Tang to vascular endothelial active substances in rats with experimental hyperlipemia and myocardial ischemia.Methods The early changes of hyperlipid and atherosclerosis were caused by utilizing multiple factors including feeding hyperlipid,propylthiouracil and high doses of VD3.Based on the above,the myocardial ischemia model was established by injecting high doses of pituitrin.The total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDLC) and low density lipoprotein cholesterol(LDLC) were measured in the 6th,12nd and 18th week respectively,and also the content of endothelin(ET),6-ketoprostaglandin F1 alpha(6-keto-PGF1α),thromboxane B2(TXB2) and angiotensin Ⅱ(AngⅡ) were determined.Results Guizhi Tang significantly improves the indexes of blood lipid metabolism,decreases TC,TG and LDLC,and increases HDLC.Guizhi Tang decreases ET,AngⅡ and TXB2 of blood serum,and increases 6-keto-PGF1α.Conclusion Guizhi Tang can regulate the function of vasoactive substance,protect endothelial cell and inhibit the formation of atherosclerosis.

Abstract:Aim To observe the role that stromal cell derived factor-1α(SDF-1α) plays in the biological functions of endothelial outgrowth cell(EOC),and the effect that rapamycin exerts on such role.Methods The mononuclear cells were harvested from umbilical cord blood by Ficoll density gradient centrifugation,induced into EOC and then expanded in vitro.The endothelial progenitor characteristics of second generation EOC were identified by immunostaining staining.The third generation EOC were devided into 4 groups,which were treated with 10 μg/L SDF-1α,10 μg/L SDF-1α+1 μg/L rapamycin,1 μg/L rapamycin and blank culture medium for 24 hours respectively.Proliferative capacity was measured by CCK-8 assay,and the migrational ability of the EOC was analysed by scarification test.Results EOC possessed many endothelial characteristics.Immunostaining showed that surface antigens factor Ⅷ,CD34 and Flk-1 were postive.The proliferative capacity and the migrational ability of the EOC were both enhanced after incubation with the 10 μg/L SDF-1α(P<0.01).While 1 μg/L rapamycin not only counteracted such effect(P<0.01),but also inhibited the biological functions of the EOC(P<0.05).Conclusoins SDF-1α can activate the proliferative capacity and the migrational ability of EOC.Rapamycin not only counteracts such effect,but also inhibits the biological functions of the EOC directly.

Abstract:Aim To study the effect of basic fibroblast growth factor(bFGF) and platelet-derived growth factor-BB(PDGF-BB) on the proliferation and migration of endothelial progenitor cells(EPC)-derived endothelial-like cells and smooth muscle-like cells.Methods EPC isolated and cultured for 5 days were stained by immunofluorescence and observed with laser confocal microscopy,the cells of which both AC133 and vWF are positive are the differentiating EPC.After 5 days of EPC culture,the medium was replaced by either control medium(DMEM,supplemented with 20% fetal bovine serum) or control medium containing rat bFGF(30 μg/L) or PDGF-BB(40 μg/L).The cultures were incubated up to 14th day.Then,endothelial cell marker(CD31 and vWF) of bFGF-induced cells(FIC) and non-induced cells(NIC) or smooth muscle cell marker(α-SMA and calponin) of PDGF-BB-induced cells(PIC) and non-induced cells(NIC) were measured by confocal microscopy.The proliferation and migration of the endothelial-like cells and smooth muscle-like cells-derived EPC were detected by MTT assays and Transwell.Results Compared with control group,EPCs induced by bFGF(named as endothelial-like cells) or EPC induced by PDGF-BB(named as smooth muscle-like cells) showed respectively more significant endothelial mark(CD31,vWF) or smooth muscle mark(α-SMA,calponin);In the period of 0～48 h,bFGF and PDGF-BB respectively promoted endothelial-like cells and smooth muscle-like cells proliferation at the corresponding concentration and there were time-dependent and dose-dependent manner.Conclusion bFGF and PDGF-BB may promote EPC to differentiate into endothelial-like cells and smooth muscle-like cells,and these endothelial-like cells and smooth muscle-like cells have the obvious ability of proliferation and migration.

Abstract:Aim To investigate the role of lipoprotein lipase(LPL) activator NO-1886 on the protein expressions of tumor necrosis factor-α(TNF-α) and IL-1β in high-fat/high-sucrose/high-cholesterol diet(HFSCD) fed miniature pigs.Methods Male,fifteen Bama-miniature pigs were randomized into three groups:control group,high-fat/highsucrose/high-cholesterol(three high) group and NO-1886 treated(HFSCD + NO-1886) group.Blood samples for plasma parameters including plasma TG and FFA were withdrawn from the orbital sinus of the animals at the end of each month following an overnight fast.The pigs were sacrificed at the end of month 5.The degree of lipid deposition in liver was examined by transmission electron microscope.The protein levels of TNF-α and IL-1beta in liver and fat tissues were measured by western blotting.Results The lipid metabolic disorder was induced in Guangxi Bama-miniature pigs by feeding high fat/high sucrose/high cholesterol diet,levels of plasma fasting TG and FFA were increased,which induced ectopic visceral lipid deposition in liver tissue and increased the protein levels of TNF-α and IL-1βin liver and fat tissues.While NO-1886 decreased the level of TG and FFA,ameliorated visceral fat deposition and reduced the protein expres-sions of TNF-α and IL-1beta in liver and fat tissues.Conclusions NO-1886 may significantly meliorate ectopic visceral lipid deposition,ameliorated lipid metabolism,decrease the protein expressions of TNF-α and IL-1β in tissues of high-fat/high-sucrose/high-cholesterol diet fed miniature pigs,and inhibit the progress of atherosclerosis.

Abstract:Aim To investigate whether ghrelin could inhibit apoptosis induced by palmiate in rat aortic endothelial cells.Methods Rat aortic endothelial cells were cultured in 0.3 mmol/L palmitate for 24 h with or without ghrelin.Cell viability was assessed by MTT assay.Apoptosis was detected using hoechst 33258 and annexin V-FITC/PI double staining assay.Spectrofluorometer assay was used to detect caspase-3 activity.Western Blot analysis was used to examine the expression of Bcl-2 and Bax.Results Exposure of cells to palmitate decreased cell viability.Palmitate increased cells apoptosis to 30.03%(P<0.01) compared with the control group(5.01%).Palmitate increased caspase-3 activity and decreased the Bcl-2/Bax ratio.While ghrelin dose dependently increased cell viability.10 nmol/L was the minimum effective level.100 nmol/L was the most significant level.Ghrelin inhibited palmitate-induced apoptosis rate to 10.03% in endothelial cells(P<0.05) compared with the palmitate group.Ghrelin decreased caspase-3 activity and increased the Bcl-2/Bax ratio at the same time.Conclusions Ghrelin inhibits palmitate-induced apoptosis.Ghrelin may be a protective factor against palmitate-induced endothelial injury.

Abstract:Aim To study the influence of probucol on the blood sugar uncontrolled type 2 diabetic mellitus patients with non-proliferative diabetic non-proliferative diabetic retinopathy(NPDR) about blood lipids,oxidative stress indicators,visual function and retinal morphology,so as to search treatment method for these patients.Methods 47 type 2 diabetes patients with 91 NPDR eyes and poor glycemic control at early stage were included.Patients were randomly divided into control and treatment groups:the control group treated by intensive therapy of blood glucose and blood pressure control,the treatment group were treated with the intensive therapy and probucol 0.375 g,2 times a day for 12 months.Before and after treatment,both groups of the patients had their blood lipids,serum level of the total antioxidant capacity(TAOC),superoxide dismutase(SOD),malondialdehyde(MDA),visual acuity,fundus,and fundus fluorescein angiography been checked.Results All 47 cases,91 eyes completed the study.Probucol obviously decreased levels of total cholesterol(TC),triglyceride(TG) and low density lipoprotein cholesterol(LDLC) in plasma of the patients.Levels of TAOC and SOD were improved significantly in the probucol group,while MDA decreased and the visual acuity improved significantly(P<0.01).The retinal capillary hemangioma,fundus bleeding and exudation,and macular edema were decreased significantly in patients of the probucol group(P<0.05).Probucol also has a role in reduction of capillary non-perfusion areas in the diabetic retina.Conclusions Probucol can not only regulate serum lipids of the poor glycemic controlled patients,but also has the action of improving antioxidant capacity.It can improve the visual function,ameliorate retinal’s microangiopathy,and decrease the incidence of macular edema.It means that probucol has a therapeutic effect in blood sugar uncontrolled NDPR patients.

Abstract:Aim To investigate the Association between endothelial progenitor cells(EPC) and the features of carotid atherosclerotic plaque.Methods Cerebral infarction patients with carotid atherosclerosis and healthy volunteers without carotid atherosclerosis were enrolled.Carotid color Doppler ultrasonography and head computed tomography(CT) or magnetic resonance imaging(MRI) were applied in all persons enrolled.According to clinical symptoms of carotid color Doppler ultrasonography,they were divided into three groups:soft plaque group,hard plaque group and control group.Blood 10 mL were drawn from ulnar vein.Mononuclear cells were isolated from peripheral blood by density gradient centrifugation and cultured on the twenty-four well fibronectin-coated cell dishes.Nonadherent cells were discarded on day 4 and culture medium was then changed.Attached cells were collected.EPCs were characterized as double positive for Dil-acLDL and FITC-UEA-Ⅰ by laser scanning confocal microscope(LSCM).EPCs adhesion,and proliferation and migration were assayed with adhesion assay,MTT assay and modified Boyden chamber assay,respectively.Results The numbers and adhesive capacity of EPCs in soft plaque group and hard plaque group were higher than those in control group.The numbers,adhesive capacity and migratory capacity of EPCs in soft plaque group were higher than those in hard plaque group.The proliferative capacity of EPCs in control group were higher than those in soft plaque group and hard plaque group.The proliferative capacity of EPCs in soft plaque group were higher than those in hard plaque group.The migratory capacity of EPCs in soft plaque group were higher than those in control group.Conclusion EPCs are associated with the features of carotid atherosclerotic plaque.

Abstract:Aim To investigate the relationship between plasma homocysteine(Hcy),methylenetetrahydrofolate reductase(MTHFR) gene polymorphisms and carotid artery plaques in the cerebral infarction patients.Methods 128 carotid system cerebral infarction patients were selected.Plasma Hcy was measured by fluorescence polarization immunoassay(FPIA).Polymorphisms of the MTHFR gene were measured by polymerase chain reaction and restriction fragment length polymorphism(PCR-RFLP).Carotid arteries were detected by carotid ultrasonography in all patients.According to ultrasonography results,the patients were divided into plaque group having carotid atherosclerotic plaque and non-plaque group.The plasma Hcy levels and MTHFR gene polymorphism were compared between two groups.Results The frequencies of TT genotype(35.8% vs 17.0%) and T allele(58.6% vs 40.4%) were significantly higher in plaque group than in non-plaque group(P<0.05).Plasma Hcy levels was significantly higher in plaque group than in non-plaque group(22.42±11.04 μmol/L vs 17.89±5.96 μmol/L,P<0.05).The Hcy concentration was significantly higher in TT genotype than CT and CC genotypes(P<0.05).Logistic regression analysis adjusted for gender,age,hypertension,diabetes,hyperlipidemia,smoking,drinking and other risk factors,plasma Hcy levels of carotid atherosclerosis remained an independent risk factor for plaque(OR 1.160(95% CI 1.034～1.301),P<0.05).Conclusions Elevated Hcy levels was an independent risk factor for carotid artery plaques in cerebral infarction patients.MTHFR C677T polymorphism was significantly related to plasma Hcy levels.

Abstract:Aim To obeserve the serum resistin expression in cerebral infarction and carotid atherosclerosis.Methods Carotid atherosclerosis of all enrolled objects were evaluated by ultrasonography and the level of resistin were detected in blood plasma by enzyme-linked immunosorbent assay(ELISA).The resistin content in cerebral infarction group was compared with control group.The relationships between resistin and carotid intima-media thickness(IMT),carotid plaque scores were analysed.Results 82.4% patients were detected to have carotid atherosclerosis in cerebral infarction group,and 36.3% patients were detected to have carotid atherosclerosis in control group.The level of resistin in cerebral infarction group was higher than in control group(23.06 ±2.47 mg/L vs 12.63±1.99 mg/L,P<0.0 1).Resistin,blood pressure,age,LDLC,high-sensitive C-reactive protein(hs-CRP) were the major risk factors of carotid atherosclerosis by Logistic.The level of resistin was positively correlated with cartiod IMT,plaque scores,LDLC and hs-CRP in plasma,and the pearson cofficient were 0.22,0.24,0.17,0.25(P<0.05).Conclusion The resistin level increases in cerebral infarction group,and the resistin may be the risk factor of carotid atherosclerosis.

Abstract:Aim To approach the short-term and long-term relative risks of ischemic stroke associated with clinically diagnosed transient ischemic attack(TIA),provide the reference frame for the prophylaxis and timely treatment of ischemic stroke.Methods We used a case-control study.1040 cases of ischemic stroke for the case group,and 2982 control subjects came from our hospital medical examination center.Clinically diagnosed TIA was ascertained from medical records.Logistic regression was used to calculate OR.Results In 1040 stroke cases and 2982 control subjects,Clinically diagnosed TIA was 187(17.98%) cases and 104(3.49%) respectively.Analysis focused on the most recent TIA before the index date.For TIA<1 month before the index date,the adjusted OR for stroke was 31.5(95%CI 11.6 to 88.4);For TIA 1 to 3 months before the index date,it was 19.2(95%CI 8.6 to 41.7);For TIA 4 to 6 months before the index date,it was 3.98(95%CI 1.38 to 7.93);And for TIA >5 years before the index date,it was 2.14(95%CI 1.22 to 2.85).Conclusions The relative risk of ischemic stroke was high for TIA diagnosed within the past 3 months,which was the critical period for prevention.

Abstract:Aim To know about the incidence rate of carotid atherosclerosis of the patients with coronary heart disease;To analyze the related risk factors of carotid atherosclerosis of the patients with coronary heart disease;To determine the relationship between the aggravation of carotid atherosclerosis and coronary atherosclerosis;To summarize the pathologic character of carotid atherosclerosis of the patients with various coronary heart disease.Methods Review the result of coronary angiography and carotid ultrasound and the related risk factors of the patients who were diagnosed as coronary heart disease through coronary angiography.The patients were divided into the carotidatherosclerosis group and the non-carotidatherosclerosis group.The patients were divided into stable angina pectoris(AP) group,unstable angina pectoris(UAP) group and acute myocardial infaction(AMI) group according to the diagnotic criterion of coronary heart disease of WHO.The patients were divided into group A(coronary artery of single vessel lession),group B(coronary artery of double vessel lession),group C(coronary artery of triple vessel lession) and group D(left main vessel lession).The aggravation of carotid atherosclerosis was graded.Results There were 198 patients with various aggravative carotid atherosclerosis among 228 patients with coronary heart disease(86.8%).Carotid atherosclerosis was much related with hypertension and non-related with age,sex,body mass index(BMI),smoking,drinking,hyperlipidermia,hyperuricermia and diabetes mellitus.The aggravation of carotid atherosclerosis got graver with the severity of coronary athero-sclerosis.But only the plaque number of group A was more than group D(P<0.05).The grading integral and Crouse integral of carotid atherosclerosis of AP group was not remarkablely less than UAP group.The grading integral of carotid atherosclerosis of AP group was less than AMI group and the Crouse integral of AP group was more.But there was no remarkable difference.The number of all plaque,and plaque of AP group was not remarkablely less than UAP group and AMI group.The plaque of all the three groups had no obvious difference.There was the plaque among the three groups.Conclusion Carotid atherosclerosis is relative to coronary heart disease.

Abstract:Aim To evaluate the correlation between the left antetior descending coronary artery(LAD) atherosclerotic stenosis and cardiac function by 64-slice spiral CT,and to discuss the mainly imaging findings.Methods 104 cases with the LAD atherosclerotic stenosis and 20 cases of controls were studied.All cases underwent 64-row CT coronary angiography,and the data were transferred to Vitrea 2.0 work station.Then,the parameters including left ventricular ejection fraction(LVEF),left ventricular end diastolic volume(LVEDV),left ventricular end systolic volume(LVESV),left ventricular myocardial mass(LVMM) were calculated.All data were analyzed with statistical software.Results There were no significant difference of cardiac functions in mild stenosis group compared with control group,and in moderate stenosis group,LVMM and LVESV increased,while LVEF decreased in a small percentage.Statistical differences of LVEF,LVEDV,LVESV,LVMM could be found in severe stenosis group compared with mild stenosis group,moderate stenosis group and the control group.Conclusion With the stenosis of the LAD increasing,the change is a dynamic process from chronic adjustment to cardiac dysfunction.64-slice spiral CT can evaluate the changes and correlations accurately.

Abstract:Aim To observe clinical treatment effect and prognosis of the emergency intervention and elective intervention treatment in acute ST-elevation myocardial infarction(STEMI) patients.Methods Clinical data of 524 patients with STEMI were retrospectively analysed,including 471 patients diagnosed in the emergency percutaneous coronary intervention(PCI) time window,who received emergency treatment,and 53 cases diagnosed beyond the emergency intervention,who received the conventional conservative treatment for 7 to 10 days before PCI.Left ventricular ejection fraction(LVEF) 7 days and six months after PCI,recurrent infarction rehospitalization within a year follow-up period,mortality rate at 30 days and a year,were observed and compared between the two groups.Results LVEF of emergency PCI group 7 days(52.3%±7.5% vs 48.2%±6.9%)and six months(54.9%±8.2% vs 50.1%±7.1%) after PCI were significantly higher than that of elective PCI group(P<0.05);Follow-up 1 year,incidence of cardiac rupture during hospitalization,recurrent infarction rehospitalization rate,mortality rate at 30 days and a year(0.42%,6.58%,3.82% and 8.28%) decreased significantly in emergency PCI group compared with elective PCI group(3.77%,16.98%,11.32% and 18.87%).Conclusions For STEMI patients,emergency interventional treatment can rapidly improve myocardial ischemia and reperfusion,then relieve symptoms of myocardial ischemia,reduce incidence of recurrent infarction and mortality.

Abstract:Atherosclerosis is thought to arise as a result of a chronic inflammatory process within the vessel wall,also the basic pathology of cardiovascular disease.Clinical studies found that pioglitazone,a peroxisome proliferator-activated receptor gamma agonist,can improve glucose and lipid metabolism,protect vascular endothelial function,reduce inflammatory markers,inhibit smooth muscle cell proliferation,anti-thrombosis,stabilize atheromatous plaque and so on,which plays a protective role in the pathological process of atherosclerosis and prevention of vascular disease,reduces the incidence of cardiovascular events and mortality.In this paper,the mechanism of the effect of pioglitazone on anti-atherosclerosis is reviewed.

Abstract:Endothelial progenitor cells(EPC) mainly derived from bone marrow are the precursor of mature endothelial cell,and have the characteristics of hematopoietic stem cells as well as the potential to differentiate into endothelial cells.EPC can participate in the process of atherosclerosis,and are closely related with cardiovascular disease.Recent studies have provided increasing evidence that gender differences exist in the morbidity and mortality of cardiovascular disease.This article tries to review the effects and mechanisms of estrogen and androgen on EPC.