Abstract:Aim To study the immunologic injury mechanism of atherogenesis by murine cytomegalovirus(MCMV) infection. Methods Animal model of atherosclerosis(C57BL/6) with apolipoprotein E knockout mice were randomized to four groups: control group,hypercholesterol diet group,MCMV infection group,MCMV infection add hypercholesterol diet group,respectively.Mice from each group were sacrificed in different period.Portions of aortas were kept in-80℃.The expression of monocyte chemoattractant protein-1(MCP-1),growth-related oncogene-α(GRO-α),fractalkine(FKN) and regulated activation normal T cell expressed and secreted(RANTES) in aortas were detected by RT-PCR and immunohistochemistry. Results In the 8th week,the expression of GRO-α in aorta tissues was significant by CMV infection,and the expression of FKN,RANTES and MCP-1 was induced.The expression of FKN was more significant in MCMV infection add hypercholesterol diet group than other groups.In the 12th week,the expression of FKN and RANTES was marked than others.Immunohistochemistry staining also showed that the expression of MCP-1,GRO-α and FKN was enhanced in atherosclerotic lesion,but expression of FKN was variable,and correlated with the atherosclerotic plaque. Conclusion MCMV infection could contribute to atherogenesis by inducing the expression of chemokines and enhance the migration of inflammatory cells.

Abstract:Aim To investigate the potential effects and molecular mechanisms of an angiotensin Ⅱtype 1 receptor blockers irbesartan on the process of atherosclerosis in high cholesterol-diet apolipoprotein E knockout(ApoE KO) mice. Methods Adult male ApoE KO mice were given normal diet or high cholesterol-diet and randomized to no treatment or irbesartan 10 mg/(kg·d) for 12 weeks.Systolic blood pressure was measured by a kind of non-invasive tail cuff system in conscious mice.The plasma total cholesterol and triglyceride concentration were measured by autoanalyzer.Atherosclerotic lesion area in aortic root was evaluated by oil red O staining.Inflammatory cytokines were measured by real-time reverse-transcription polymerase chain reaction(PCR) and Western blotting. Results The ApoE KO mice with high cholesterol-diet was associated with a marked increase in plasma lipid levels,atherosclerotic lesion area,as well as the expressions of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),monocyte chemoattactant protein-1(MCP-1) and vascular cell adhesion molecule-1(VCAM-1).These changes were suppressed in mice that were treated with irbesartan 10 mg/(kg·d) for 12 weeks concomitant with high cholesterol-diet administration,with no significant change in systolic blood pressure and plasma lipid levels. Conclusion The results suggest that irbesartan can attenuate atherosclerosis,and this effect is partly related to the inhibition of inflammatory response which leads to the decrease in the expression of inflammatory cytokines.

Abstract:Aim To observe the changes of serine/threonine kinase 2(Akt 2)protein expression in pulmonary arteriy smooth muscle cells(PASMC) induced by hypoxia in rats,and to investigate the value of Akt 2 signaling pathway in hypoxia pulmonary hypertension(HPH). Methods The pure PASMC was cultured by tissue-sticking methods.Akt 2 protein expressions were determined by Western blotting after PASMC were exposed to hypoxia for 2 h,8 h,12 h and 24 h respectively.The changes of PASMC proliferation were determined by MTT and 3 H-TdR incorporated way. Results The value of 3 H-TdR was 0.372±0.059 under normoxia.And accompanying the prolongation of hypoxia time,the value of 3 H-TdR kept heightening.It was 0.703±0.100 in 12 h and significantly different from normoxia group;The value of MTT was 8374.39±545.31 under normoxia.And accompanying the prolongation of hypoxia time,the value of MTT kept heightening.It was 11208.35±678.82 in 24 h group and significantly different from normoxia group.The protein of Akt 2 could be detected in all groups,and the expressive level of protein showed a possible relationship with PASMC prolferation. Conclusion The results suggest that Akt 2 signaling pathway may play an important role in the hypoxia pulmonary hypertension.

Abstract:Aim To determine if treatment with hydroxymethylglutaryl-coenzyme A reductase inhibitors(statins) can influence the development of experimental abdominal aortic aneurysms(AAAs). Methods Twenty-four Wistar Rats were randomly divided into 2 equal groups(n=12).An animal model of AAAs was established by perfusion of the aorta with elastase,treated for 14 days with Atorvastatin 4 mg/(kg·d) or with NS alone.The expression of MMP-9 and NF-κB were observed in aortic tissue by immunohistochemistry staining or by RT-PCR. Result The incidence of AAAs was reduced by 70%(2 of 10 rats treated with atrovastatin vs.9 of 10 rats treated with vehicle ;P<0.05).The development of AAAs in vehicle-treated rats were accompanied by a dense transmural inflammatory response,and complete destruction of the elastic media.MMP-9 and NF-κB expression increase significantly in vehicle-treated rats.Inflammatory infiltration was less found in Atorvastatin-treated rat,and little MMP-9 and NF-κB expression in atrovastatin-treated rats. Conclusion Treatment with Atorvastatin reduced the development of elastase-induced experimental AAAs.The underlying mechanism was associated with structural preservation of aortic wall elastin and the presence of a chronic inflammatory response,there was a relative reduction in aortic wall expression of MMP-9and NF-κB,the effect is independent on serum cholesterol levels.

Abstract:Aim To observe the hyperlipemia rabbit hemodynamic changes in the early stage of atherosclerosis by using Doppler ultrasound at different time point. Methods 60 white big-ear rabbits were divided into two groups randomly: 15 rabbits(control group) were fed with normal rabbit chow,while 45 rabbits(experiment group) were fed with high cholesterol diet.Then each group was divided into three subgroups that were fed for 4 weeks,8 weeks and 12 weeks respectively.Hemodynamic parameters(including systolic peak velocity,diastolic velocity,pulsatility index,resistent index,diameter of systole and diastole),blood lipid levels and the pathological examination were detected at each time-point. Results Levels of cholesterol and low-density lipoprotein cholesterol of experiment group increased significantly and reached peaks at 8 week,then they decreased slightly at 12 week;levels of high-density lipoprotein cholesterol decreased significantly from 8 week;triglyceride level had an obvious elevation during the experiment(P<0.01).The rabbit blood vessel diameter and resistance index were with significant difference compared with control group at the same time-point(P<0.01).Systolic peak velocity of 12 week,diastolic peak velocity of 4 week and 8 week,and pulsatility index of 4 week and 12 week of experiment group were all significant in statistics(P<0.05).Among the subgroups Dd had no significant changes;Ds changed at 4 week and 12 week(P<0.05);Vs,Vd and PI between 4 week and 12 week,8 week and 12 week were different(P<0.01 or P<0.001);RI between 4 week and other two time points,8 week and 12 week were different(P<0.01). Conclusion Hemodynamic changes of color Doppler,that presented earlier than morphology,may dynamically detect the progression of atherosclerosis,and play an important role in early diagnosis of atherosclerosis.

Abstract:Aim To investigate the effects of C-reactive protein(CRP) on the proliferation and chemotaxis of endothelial progenitor cells(EPC) of human peripheral blood. Methods Mononuclear cells were isolated from human peripheral blood and cultured for 7days.Attached cells were incubated with different concentration of CRP(1.0 mg/L,2.5 mg/L and 5.0 mg/L) for different times(24,48 and 72 h).MTT assay and quantified colony forming units(CFU) were used to assess the proliferation of EPC after treated with CRP.Meanwhile,chemotaxis assay was used to quntified EPC induced by VEGF.NO in the supernatant was measured by nitrate reductase assay.RT-PCR was employed to detect mRNA expression of endothelial nitric oxide synthase(eNOS). Results Incubation of EPC with CRP decreased the number and the function of EPC. Conclusions It is suggested that CRP can promote endothelial dysfunction by means of depressant EPC proliferation and migratory.

Abstract:Aim To investigate the effects of CD40 ligand from activated platelets on the expression of tissue factor and tissue factor pathway inhibitor in human vascular endothelial cells(hUVEC) and its molecular mechanism. Methods Activated platelets and human umbilical vein endothelial cells were co-incubated in the same systerm,and then the expression of tissue factor and its inhibitor in endothelial cells and the effects of blocking CD40-CD40L signal pathway on their expression were observed. Result After 8 hours of the co-incubation of activated platelets and human umbilical vein endothelial cells,tissue factor mRNA and its protein expression level in hUVEC were raised.Nevertheless these effects could be attenuated by CD40L monoclonal antibody and CD40 monoclonal antibody added into media before platelets being induced(P<0.05),but tissue factor pathway inhibitor mRNA expression level in hUVEC was not changed. Conclusions Activated platelets could induce the expression of tissue factor in endothelial cells by the expression of CD40L, but could not enhance the expression of tissue factor pathway inhibitor.This indicates that these effects may play a promoting role in unstable plaque formation and the development of atherosclerosis.

Abstract:Aim To investigate the change rule and correlation of heme oxygenase-1(HO-1)/ carbon monoxide(CO) and nitric oxide synthase(NOS) / nitric oxide(NO) and the influence of amlodipine on the two systems in atherosclerotic progress. Methods The rabbits received 1% cholesterol diet(n=16) for eight weeks;after the eight weeks,one group(n=8) were fed with normal diet for eight weeks and onther group(n=8) were administrated with amlodipine. Results Compared with contrast group,in model group the levels of serum lipids and plasma carbon monoxide were increased obviously(P<0.01);however,the levels of serum nitric oxide and the expression of heme oxygenase-1 and inducible nitric oxide synthase in aortic were decreased markedly(P<0.01).Compared with model group,in amlodipine group the levels serum lipids and serum nitric oxide were no significant effects(P>0.05),the levels of plasma carbon monoxide were decreased obviously(P<0.01),while the expression of heme oxygenase-1 and inducible nitric oxide synthase in aortic reduced greatly(P<0.01). Conclusion In atherosclerotic progress,between heme oxygenase-1(HO-1)/carbon monoxide(CO) and nitric oxide synthase(NOS)/nitric oxide(NO) system appears the reciprocal relationship,and amlodipine may delay atherosclerotic progress by dowhregulate the two systems.

Abstract:Aim To observe the expression of found in inflammatory zone1(FIZZ1) in aortic atherosclerosis lesion and the interventional effect of rosiglitazone in apolipoprotein E-knockout mice. Methods Eight-week-old apoE knockout mice were divided into atherosclerosis(As) group(n=10) and rosiglitazone group(n=10).Wildtype C57/BL mice(n=10) were used as normal control group(n=10).All mice were fed with normal chow diet.In addition to normal diet,rosiglitazone group received rosiglitazone 10 mg/(kg·g) of body weight.After 12 weeks,aorta were used for histomorphometric analysis of atherosclerosis lesion,immunohistochemistry and RT-PCR for the expression of FIZZ1.Vessel bloods were collected for plAsma lipid and high sensitive-CRP(hs-CRP). Results There is no significant difference in blood lipid level between As group and rosiglitazone group.The hs-CRP level in As group is significantly higher than that in rosiglitazone group.Histomorphometric analysis showed that the area of atherosclerosis plaque in As group was significantly bigger than that in rosiglitazone group.Immunohistochemistry and RT-PCR showed that the expression of FIZZ1 in As group were higher than that in rosiglitazone group. Conclusion The expression of FIZZ1 in atherosclerosis lesion was observed.Rosiglitazone can decrease hs-CRP level,reduce the expression of FIZZ1 in aortic atherosclerosis lesion.It is not related to modulation of blood lipid that rosiglitazone inhibits the development of aortic atherosclerosis.Anti-inflammation is a potential mechanism.

Abstract:Aim To study the effect of alendronate on the expression of osteoprotegerin(OPG) in rat aorta,and investigate the possible mechanism of inhibiting artery calcification by alendronate. Methods Eighteen 4-week-old male SD rats were randomly divided into the control group(n=6),calcification group(n=6) and alendronate group(n=6).The later two groups were subcutaneously injected with warfarin(150 mg/kg per 12 hours for 5 days) and Vitamin D3[3 MU/(kg·d) for 3 days] to induce extensive calcification of the aorta.Alendronate group was treated with daily subcutaneous injection of alendronate 4 days before the first warfarin dose.The control group was given daily subcutaneous injection of 0.9%NaCl.Thoracic aortas were exercised for analysis of calcification and OPG expression using histomorphometry,spectrophotome RT-PCR and immunohistochemistry. Results The large area of focal darkly stained regions by Von Kossa staining reduced obviously in alendronate group.The level of calcium content in aorta wall was significantly lower in the alendronate group than in the calcification group(67.33±19.86 μmol/g vs 114.3±23.23 μmol/g,P<0.05).The relative content of OPG mRNA in aortic wall was higher in the alendronate group than in the calcification group(P<0.01).The similar results was drawn by immuohistochemistry.OPG protein expression was similar to that for its mRNA expression. Conclusions Alendronate inhibited artery calcification by upregulating the expression of OPG.

Abstract:Aim To study the effect of united therapy of probucol(P) and clopidogrel(Anti-thrombosis,A) and atorvastatin(statin,S) on rabbit atherogenesis. Methods 36 rabbits were randomized into four groups.The one group did not receive treatments and served as control group.Atherosclerosis was induced in the aorta arteries of rabbits by atherogenic diet in the next groups for 6 weeks.Then, animals were randomized to receive either no treatment or therapy.One group of treatment were received atorvastatin 2.5 mg/(kg·d),and the other accepted probucol(1.0 g/d) and clopidogrel 5 mg/(kg·d) and atorvastatin 2.5 mg/(kg·d) and killed after 4 weeks. Results Atorvastatin and united therapy induced a significant reduction in serum lipids and in lesion size,also were the area and thickness in neointima.Arterial macrophage infiltration was abolished by the treatment.Notedly,there was a more evident effect in united therapy than Atorvastatin in anti-atherosclerosis. Conclusions Atorvastatin and united therapy diminish the plaque lesion and neointimal inflammation in rabbit atherosclerosis model,and the effect of united therapy was more than atorvastatin.This could contribute to the more stabilization of the atherosclerotic plaque,and may be a more meaning additional prediction for the reduction of acute ischemic events in patients treated with united therapy.

Abstract:Aim To elucidate the mechanism of advanced glycation end products(AGE) inducing oxidative effects in cultured ECV304 cells. Methods ECV304 cells were cultured in vitro with AGE-human serum albumin(HSA),or pretreated with Apocynin or GF109203 or Genistein for 0.5 h,then cultured with AGE-HSA.After 1 h,the level of O2-· was measured with cytochrome C,the level of reduced glutathione was measured with ThioGlo-1. Results O2-· increased from 1.37±0.67 nmol/(107·h) to 3.44±0.40,10.67±0.67 and 10.93±0.67 nmol/(107·h),and reduced glutathione decreased from 9.54±0.41 nmol/106 to 9.02±0.21,8.41±0.34,and 8.02±0.18 nmol/106 after 12.5 mg/L,50 mg/L,200 mg/L AGE-HSA stimulating;Apocynin,GF109203 and Genistein could inhibit these effects. Conclusion AGE-HSA could activate NADPH oxidatiate enzyme via protein kinase(PKC) and tyrosine protein kinase(TPK)to induce O2-· increasing and reduced glutathione decreasing and enhance intracellular oxidative effects.

Abstract:Aim To compare efficacy and safety of clopidogrel two treatment programs on preventing cardiovascular events after percutaneous coronary intervention. Methods 245 cases of patients undergoing coronary intervention were analyzed retrospectively.The patients of the observation group(120 cases) were given double of capacity clopidogrel 300 mg and 225 mg on the preoperative 48 h and 24 h respectively,then conventional oral 75 mg,qd;and the patients of the control group(125 cases) were given clopidogrel 300 mg on the preoperative 6 h single,then conventional oral 75 mg,qd.After three months,incidence rates of cardiovascular events,thrombosis and bleeding complications were compared between the two groups. Results After three months of treatment,blood coagulations of the two groups were similar(P>0.05).The incidence of reischemic event and thrombosis in observation group was significantly lower compared with the control group(P<0.01),but the incidences of bleeding of the two groups had no significant difference(P>0.05).Nobody was obliged to stop treating for drug side-effects. Conclusion giving double load of clopidogrel to patients with coronary heart disease undergoing coronary intervention before operation can reduce the incidences of reischemic event and thrombosis,and is safe.

Abstract:Aim To investigate the relationship between plasma soluble thrombomodulin(sTM) with coronary artery disease(CAD) and explore its association with other risk factors of CAD in 74 male patients. Methods Plasma sTM was measured by enzyme linked immunosorbent assay(ELISA) in all subjects,including 50 patients with CAD and 24 controls.All the subjects were documented by coronary artery angiography.Clinical information,such as age,body mass index,homocysteine,etc,was analyzed. Results The plasma sTM in CAD patients(32.9±9.4 μg/L) was significantly higher than that in the controls(24.6±5.1 μg/L,P=0.0001).Univariate Logistic regression showed that sTM was a significant risk factor for CAD.Oneway analysis of variance showed the patients in different clinical severities had different sTM levels(P=0.000) and patients with acute myocaidial infarction(AMI) or unstable angina(UA) owned a higher sTM level than the patients with stable angina(SA) or controls.If all subjects were assigned to quartiles based on their sTM values and OR value was estimated by comparing the likelihood of different severities of CAD among the quartiles,the OR value of developing more serious CAD augmented 2.5-fold for each increasing quartile of sTM in an ordinal logistic regression.Likewise,and the OR value of developing one rank or more than one rank in Gensini' scores was 1.89 for each increasing quartile of sTM.Plasma sTM levels were only related to age,high sensitive C-reaction protein (hs-CRP) and D-dimer significantly among those variables.Furthermore,Gensini' scores correlated with sTM significantly(r=0.43,P=0.001). Conclusions As a maker for endothelial damage,the sTM level is closely related to the severity of CAD and the severity of diseased coronary arteries in male patients.

Abstract:Aim To provide reference for the therapy and the prevention of cerebrovascular disease,we investigated the relationship between age and the location and the severity of stenosis in ischemic stroke patients. Methods From January 2004 to October 2005,311 patients with ischemic stroke were selected and diagnosed by DSA.Among them,there were 223 males and 88 females ranging from 18 to 82 years old.They were divided into three groups by age: young group(<45 years old),middle-age group(45～59 years old),and elder group(≥ 60 years old).Artery stenosis was diagnosed based on the North American Symptomatic Carotid Endarterectomy Trial(NASCET).This study examines the relationship between age and the location and severity of stenosis. Results ① 218(70.1%) of 311 patients with ischemic stroke suffered from artery stenosis.The incidence of ischemic stroke increases with age.The incidences between genders were not significantly different(P>0.05).②The distributions of ischemic stroke differs in different age groups(P<0.01): the young group exhibits high incidence in the intracranial artery(69.0%),but the middle-age and elder groups exhibit high incidences in both the intra and extracranial arteries(respectively,36.2% and 38.2%).③The numbers of intra-and extracranial artery stenosis were compared among the different age groups—while the difference for extracranial artery was significant(P<0.05),the difference for intracranial artery was not(P>0.05).④Among the different age groups,the severity of stenosis was not significantly different(P>0.05). Conclusions In Chinese patients with ischemic stroke,the distribution of artery atherosclerosis is increasing with age,but not necessarily more severe.The severity of stenosis is also related to the diameter of the blood vessel.

Abstract:Aim To detect the association of estrogen receptor(ER)α gene PuvⅡ polymorphisms with severity of coronary heart disease(CHD) of Han nationalities in South China. Methods By using PCR-RFLP analysis method,ER gene types in 236 CHD patients and 117 healthy controls were detected,and the association of gene types with severity of CHD and number of involved vessel and Gensini coronary score were compared. Results The frequencies of PP,Pp and pp types of PvuⅡ polymorphism in CHD patients and controls were 19.9%,44.5% and 35.6% vs 9.4%,48.7% and 41.9%,the frequency of PP genotype in CHD was significantly higher than that of control(P<0.05).There were no significant differences in the distribution frequency of three genotypes and two alleles of PvuⅡamong subgroups with different severity of disease(P>0.05),the score in PP homogeneous was significantly higher than those in Pp genotype and pp homogeneous(P<0.05). Conclusions The ER-α gene PvuⅡ polymorphism is associated with CHD and severity of CHD in Han nationality of South China.

Abstract:Aim To investigate the influences of probucol combined with atorvastatin and solo atorvastatin on success rate of lipid and prognosis in acute coronary syndrome(ACS) patients at 1 month. Methods Ninety ACS patients were assigned to solo atorvastatin group(20 mg/d atorvastatin for 1 month,n=49) or therapeutic alliance group(20 mg/d atorvastain and 1.0 g/d probucol for 1 month n=41) in a randomized block,controlled study.Serum levels of blood lipids were detected before and aftert teatment.Then prognosis was compared in the two groups. Results The success rate of blood lipids of therapeutic alliance group was higher than solo atorvastatin group,and the prognosis was insignificantly between the two groups. Conclusion Both atorvastatin and probucol may cut down blood lipids in the ACS patients,and atorvastatin combined with probucol can elevate the success rate of blood lipids.