Abstract:Aim To investigate the influence of icariin on adhesion molecule gene expression in hyperlipidemia rats.Methods 40 male healthy SD rats were randomly divided into four groups:control group,high cholesterol group,high cholesterol with low dose icariin(100 mg/kg)group,high cholesterol with high dose icariin(200 mg/kg)group.Serum lipid level of all rats was measured and the aorta was taken for pathologic analysis.The mRNA expression of both intercellular adhesion molecule-1(ICAM-1) and vascular cell adhesion molecule-1(VCAM-1) on the aorta was analyzed by realtime PCR.Results As compared with the high cholesterol group,the level of total cholesterol(TC),triglyceride(TG) and low density lipoprotein cholesterol(LDLC) in high cholesterol with icariin was obviously lower(P<0.05 or 0.01);the mRNA expression of ICAM-1 was significantly decreased(P<0.01);but the mRNA expression of VCAM-1 was not significantly inhibited.Conclusion Icariin can reduce the level of blood lipid and the mRNA expression of ICAM-1 in hyperlipidemia rats.

Abstract:Aim To explore the change of nitric oxide in human umbilical vein endothelial cells induced by different dose of formaldehyde and the mechanisms of the nitric oxide change.Methods Human umbilical vein endothelial cells(hUVECs-12) were cultured in dulbecco's minimum essential medium(DMEM) containing 10% fetal bovine serum.At 80% confluence,cells were washed with D-Hank's fluid and cultured in DMEM containing 1% fetal bovine serum for 24 h.Cells were incubated in the presence or absence(control) of formaldehyde.In order to test the dose-effect of formaldehyde-induced endothelial cells change of nitric oxide,the cells were treated with formaldehyde at the dose of 0.0 μmol/L and 5,10,20,40,80 μmol/L for 24 h respectively.To explore the injuring mechanism of formaldehyde on endothelial cells,hydrogen peroxide(100 μmol/L),a potent free radical,was selected as a positive control;Vitamin C(100 mg/L),an anti-oxidant medicine,was selected as a protective group.After 24 h,the transudation content of lactate dehydregenase was observed by colorimetry reaction,the level of malondialdehyde in supernatant-conditioned media of cells was measured by thiobituric acid reaction(TBA) method,the level of nitric oxide in the culture medium was observed to evaluate the influence feature of formaldehyde by nitrate reductase method,the nitric oxide synthase activity was measured by biochemical method,the protein and mRNA level of inducible nitric oxide synthase and endothelial nitric oxide synthas were examined by immunohistochemistry and RT-PCR approach respectively to elucidate the injuring mechanism.Results Formaldehyde conspicuously inhibited the activity of Endothelial nitric oxide synthas,expression of endothelial nitric oxide synthas protein and endothelial nitric oxide synthas mRNA.Formaldehyde induced the activity of inducible nitric oxide synthase,expression of inducible nitric oxide synthase protein and inducible nitric oxide synthase mRNA,and increased the production of nitric oxide,malondialdehyde and the transudation content of lactate dehydrogenase on Human umbilical vein endothelial cells in dose-dependent manner;Vitamin C inhibited the activity of inducible nitric oxide synthase,expression of inducible nitric oxide synthase protein and inducible nitric oxide synthase mRNA,and increased the activity of endothelial nitric oxide synthas.Conclusion Formaldehyde can conspicuously cause change of nitric oxide on human umbilical vein endothelial cells when the concentration is between 5 and 80 μmol/L.This change is related to reducing the activity and expression of endothelial nitric oxide synthas,and increasing the activity and expression of inducible nitric oxide synthase.

Abstract:Aim To investigate the feasibility of differentiation of bone marrow mesenchymal stem cell(BMSC) into smooth muscle cell(SMC) in vitro under platelet-derived growth factor-BB(50 μg/L) and high concentrations of fetal bovine serum(FBS)(20%),and the positive effect of myocardin on BMSC differentiating into SMC.Methods Mouse BMSCs were isolated and purifed from bone marrow,then 50 μg/L PDGF-BB and FBS(20%)were added to the BMSCs.And the cells were cultuted for 4 and 8 days respectively.Immunohistochemistry assay and image analysis were used to value the exptession extent of α-smooth muscle actin(α-SMA),smooth muscle myosin heavy chain(SM-MHC)and myocardin.Results Intensities of the three protein immunostaining significantly increased after 4 and 8 days.And Test of One-way ANOVA showed that there was statistically significance in different times(P<0.05).Conclusions BMSC could be induced to differetiate into smooth muscle-like(SM-like) cells through adding PDGF-BB(50 μg/L) and FBS(20%) with BMSC in vitro.Myocardin may play an important role in this process.

Abstract:Aim To observe the effects of lysophosphatidic acid(LPA) on system generation of adrenomedullin(ADM) with its receptors:calcitonin receptor like receptor(CRLR) and receptor activity modifying protein(RAMP),and the effects of ADM on proliferation of the vascular smooth muscle cells(VSMC) induced by LPA.Methods VSMC of rat aortic were cultured by explant method.DNA synthesis of VSMC were measured by 3H-TdR incorporation.The ADM concentration in VSMC were determined by radioimmunoassay.Results LPA stimulated the production of ADM on vascular smooth muscle cells in rats.LPA up-regulated the expression of mRNA of ADM and its receptors CRLR,RAMP2 and RAMP3 in VSMC.ADM can inhibit 3H-TdR incorporation of rat VSMC stimulated by LPA.ADM inhibited activation of mitogen-activated protein kinase(MAPK) in VSMC induced by LPA.Conclusions ADM and its receptors inhibit VSMC proliferation which involves in its inhibition on MAPK activity.

Abstract:Aim To investigate expression of farnesoid X receptor(FXR) in murine macrophage cell line ANA-1 and evaluate the effect of FXR agonist chenodexycholic acid(CDCA)on the expression of monocyte chemoattractant protein-1(MCP-1) in ANA-1 cells.Methods ANA-1 cells were cultured and treated with FXR agonist CDCA.The mRNA level of FXR and MCP-1 were detected by reverse transcription-polymerase chain reaction(RT-PCR).Western blot was used to assess the expression of FXR and MCP-1 protein level.Results FXR expressed in ANA-1 cells and acted in an autoregulatory fashion.CDCA significantly reduced MCP-1 mRNA and protein level in ANA-1 cells(P<0.05).Conclusions FXR experss in murine macrophage cell line ANA-1.After activated by CDCA,FXR may downregulate MCP-1 directly or indirectly.

Abstract:Aim To investigate the effect of Ezetimibe on cholesterol accumulation in loaded-lipid cells derived from vascular smooth muscle cells(VSMC).Methods Loaded-lipid cells model were established by 20 mg/L Chol∶MβCD treated VSMC.Loaded-lipid cells were respectively treated with different concentration of ezetimibe(0,3,10,30 μmol/L) for 24 h,and with 30 μmol/L of ezetimibe for different time(0,6,12,24,48 h).The concentration of total cholesterol(TG) and free cholesterol(FC) were detected by high performance liquid chromatography(HPLC).Cholesterol accumulation in cells were observed by oil red O staining.Results(1) Intra-cellular TG were110.8±3.56,90.5±.2.30,60.9±1.86 mg/g and FC were 85.5±3.12,74.6 ±2.89,53.8±1.56 mg/g,respectively with loaded-lipid cells treated with different concentration of ezetimibe.Compared with model group,there was significant difference in medium-dose and high-dose groups(P<0.05),TG and FC had a reducing trend with rising drug concentration(tendency χ2=11.56,P=0.007) and the best drug concentration was 30 μmol/L.(2)Treated loaded-lipid cells for different time(0,6,12,24,48 h) with 30 μmol/L ezetimibe,the content of TG were 120.1±2.37,117.9±3.05,80.3±.2.81,69.2±1.56,72.1±2.98 mg/g and FC were 92.9±2.11,90.7±3.76,71.4±2.57,60.1±1.21,64.2±2.48 mg/g respectively.TG and FC gradually decreased with treated time,and both of them had negative correlations with treated time(r=-0.776,P<0.05 and r=-0.786,P<0.05 respectively).Compared with 0 h group,there was significant difference for FC in 12 h,24 h and 48 h groups(P<0.01).There was time-depended tendency for ezetimibe and it was best for 24 hours.Conclusion Ezetimibe decrease cholesterol accumulation in vascular smooth muscle cells.

Abstract:Aim To investigate the expression of matrix metalloproteinases(MMP) and protein kinase C(PKC) in aorta of atherosclerotic rats and effect of atorvastatin on them.Methods Fifty female Sprague-Dawley rats were randomly divided into normal diet group(n=10,control group);vitamin D3 injection and high cholesterol diet group(n=40).After 8 weeks,vitamin D3 injection and high cholesterol diet rats were randomly switched to receive atorvastatin [5 mg/(kg·d)](n=20,atorvastatin group) or normal diet(n=20,model group).Another eight weeks later,all the rats were killed and some of their aortas were observed by light and electron microscope,the left aortas were removed for western blot analysis to detect MMP-2,MMP-9 and PKC;at the begin and end of experiment,serum was collected for serum lipid determining.Results There were no significant changes in serum lipids at the beginning of experiment between the groups,but after two months,cholesterol,low density lipoprotein(LDL),triglyceride(TG) of atorvastatin group were significantly lower than those of model group(all P<0.01),but higher than control group(all P<0.01).The pathologic changes of atorvastatin group were less serious than those of model group,there showed no any pathological changes in control group.MMP-2,MMP-9 and PKC were significantly higher in the model group than those in the control group(all P<0.01),But compared with the model group,MMP-2,MMP-9 and PKC levels were markedly lower in the atorvastain group(all P<0.01).Conclusions Atorvastatin inhibiting PKC expression may play an important role in depressing the express of MMP,stabilizing the atherosclerotic plaque.

Abstract:Aim To investigate the effects of captopril on vascular endothelial dysfunction induced by homocysteine thiolactone in rat in vivo.Methods The model of hyperhomocysteinemia(HHcy) in rats was induced by intragastric gavaged HTL,and the intervention groups were intragastric gavaged captopril in water.After treatment of 8 weeks,endothelium-dependent relaxation(EDR) of aortic rings were examined.Activity of paraoxonase1(PON1) and angiotensin-converting enzyme(ACE) in serum were analyzed.The masculine cell ratio of NF-κB P65 were detected by immunohistochemistry in vascular of rats.Results HTL significantly inhibited EDR induced by ACh,induced the activation of NF-κB P65 of vascular endothelium cell in rats;degraded the activity of PON1 and SOD in serum;down-regulated the PON1mRNA in the liver.Captopril could improve the EDR response and maintain the activity of PON1;up-regulated the PON1mRNA in the liver in dose-dependent manner.Captopril simultaneously inhibited the activation of NF-κB P65 of vascular endothelium cell in rats.Conclusion Captopril could protect vascular endothelial function induced by homocysteine thiolactone in rats.

Abstract:Aim To elucidate the relationship between the apolipoproteins,especially apoA-I and the alteration of HDL subclasses in type Ⅱa and Ⅱb hyperlipidemic subjects.Methods Apolipoprotein(apo) A-I contents of serum HDL subclasses in 68 type Ⅱa and 67 type Ⅱb hyperlipidemic subjects and 233 normal controls were determined by two-dimensional gel electrophoresis coupled with immunodetection method.Results Compared with normal controls,the contents of small-sized preβ1-HDL,HDL3b and HDL3a were significantly higher(P<0.01),but those of large-sized HDL2a and HDL2b were significantly lower(P<0.01 or P<0.05) in type Ⅱa and Ⅱb hyperlipidemic subjects.In addition,preβ1-HDL and HDL3a increased significantly(P<0.05) while HDL2a and HDL2b decreased significantly(P<0.01) in type Ⅱb versus type Ⅱa hyperlipidemic subjects.Both in type Ⅱa and Ⅱb hyperlipidemic group,compared to the subjects with high apo A-I levels,all small-sized HDL subclasses(except preβ2-HDL)increased significantly(P<0.01 or P<0.05) while large-sized decreased significantly(P<0.01) in the subjects with low apo A-I levels.Correlation between HDL subclasses and apolipoproteins showed that apoA-I was positively and significantly correlated with all HDL subclasses(P<0.01).In contrast,concentrations of apoB100,apoCⅢ showed positive and significant correlation with small-sized preβ1-HDL while negative and significant correlation with large-sized HDL2a and HDL2b.Conclusions The particle size of HDL shifted towards smaller size in type Ⅱa and Ⅱb hyperlipidemic subjects,and the shift was more prominent in type Ⅱb hyperlipidemic subjects.Apolipoprotein levels,especially apoA-I level influence the distributions of HDL subclasses in type Ⅱa and Ⅱb hyperlipidemic subjects.

Abstract:Aim To study the sensitivity of augmentation index(AI) and pulse way velocity(PWV) in evaluation of aortic stiffness.Methods Seventy-three continuous ambulatory peritoneal dialysis(CAPD) patients in a single center were included in the present study.Ankle-brachial pressure index(ABI) was measured.The patients were classified into ABI≤0.9 group and ABI>0.9 group.Aortic stiffness was assessed by brachial pulse pressure(PP),central augmentation pressure(C-AP),AI,central PP(C-PP) and carotid-femoral pulse wave velocity(CF-PWV).Results The CF-PWV in ABI≤0.9 group was higher than that of ABI>0.9 group.However,there was no difference in AI between the two groups.CF-PWV was positively correlated with ABI(P<0.001),but AI was not correlated with ABI.Conclusions AI was not as sensitive as PWV in evaluating aortic stiffness.Therefore,we should be cautious in utilizing AI to evaluate aortic stiffness.

Abstract:Aim To investigate the value of rest 99m Tc-MIBI myocardial single photon emission computed tomography(SPECT) in diagnosis of old myocardial infarction.Methods Vector cardiogram(VCG),ultrasonic cardiography(UCG) and rest 99m Tc-MIBI SPECT were performed in the 25 patients with dubious old myocardial infarction(22 inferior and 3 anterior myocardial infarction).Positive rate of rest 99m Tc-MIBI SPECT myocardial imaging were compared with VCG and UCG for diagnosis of old myocardial infarction.Results In diagnosis of old myocardial infarction,positive rate of rest 99m Tc-MIBI SPECT was 88%,and were significantly higher than VCG(32%) and UCG(24%;P<0.05).In diagnosis of old inferior myocardial infarction,positive rate of rest 99m Tc-MIBI SPECT(82%) were significantly higher than VCG(27%) and UCG(23%;P<0.05).Consistent rate of rest 99m Tc-MIBI SPECT with angiography was higher than the VCG(100% vs 25%).Conclusion In diagnosis of old myocardial infarction especially inferior myocardial infarction,rest 99m Tc-MIBI SPECT myocardial imaging has significant clinic value.

Abstract:Aim To evaluate the therapeutic effect of sodium ozagrel on treatment of transient ischemic attack(TIA).Methods 70 cases of TIA patients were allocated randomly into treatment group and treated control group(35 cases in every group).The treatment group was given 80 mg of sodium ozagrel intravenously twice a day and Ligustrazine 80 mg intravenously once a day,the treated control group was given 80 mg Ligustrazine once a day,both groups were given 5 000 u low molecular weigh heparin subcutaneously twice a day and 100 mg asprin once a day.Carotid internal-media thickness(IMT) and arterial plaque of all patients and normal controls were measured by carotid ultrasonography.The platelet aggregation rate and fibrinogen level were observed in both treatment groups before and after 14 d treatment.The cerebralvascular events were observed in 30 d.Results The difference of fibrinogen level and platelet aggregation rate in the treatment group was significant compared with the treated control group(P<0.05).The difference of IMT and percentage of carotid arterial plaque in the 70 cases of TIA was significant compared with the normal controls(P<0.01).Compared with the treated control group,the cerebralvascular events in the treatment group decreased(P<0.01).Conclusion Sodium ozagrel is effective and safe in treatment of TIA and progress the prognosis.

Abstract:Aim Discuss the relation between vascular endothelium function,body composition and glucose metabolism in middleaged patients with impaired glucose tolerance.Methods Adopt the Philips IU-22 color Doppler ultrasound diagnostic system to detect vascular endothelium function,body composition and glucose metabolism indexes,and process a statistic analysis.Result(1) The rate of impaired glucose tolerance group to catch coronary heart disease,the positive rate of diabetes family history and family history in cardiovascular disease,are all obviously higher than the normal glucose tolerance group.(2)Compared to the normal glucose tolerance group,the BMI,BF%,waist circumference and 2-hour postprandial blood glucose of the impaired glucose tolerance group show significant difference,while the WHR and Fasting blood glucose are obviously higher.There is no significant difference in the SBP and DBP between two groups.(3)There is significant difference in the brachial arterial Dia-P% between two groups,while the brachial arterial internal diameter and Dia-N% show no significant difference.(4)The Dia-P% of middleaged impaired glucose tolerance patients is related the OGTT2h blood glucose.(5) The multivariate successive regression equation of Dia-P% in the middleaged impaired glucose tolerance patients show a predictor variable:OGTT2h blood glucose.Conclusion Brachial arterial vasodilation dysfunction exists in the middleaged impaired glucose tolerance patients,and the indexes of body composition and glucose metabolism change.Meanwhile,the abnormality of insulin resistance and OGTT2h blood glucose play an important role in the brachial arterial vasodilation dysfunction of middleaged IGT patients.

Abstract:Aim To evaluate the prevalence of atherosclerotic renal artery stenosis(ARAS) in patients undergoing selective coronary angiography,determine the relationship of ARAS and coronary heart disease(CHD),and to investigate the related factors of ARAS.Methods A total of 410 patients with suspected CHD underwent selective renal artery angiography immediately after coronary angiography.Independent sample test and multivariate logistic regression analysis were used to study the independent risk predictors of ARAS.Results 88(21.4%) patients observed with ARAS were included 7.3% in early,9.7% in mediate and 4.3% in severity.The incidence of ARAS in 285 patients with CHD was 26.3%,and it was higher than in patients without CHD(10.4%,P<0.05).130 renal arteries were narrow more than 30% of lumen diameter,with 61.5% in the ostial,36.9% in the main stem,and 3.8% in the branch of renal artery.The multivariate logistic regression analysis released that the important related factors were female,hypercholesterolemia,lower high density lipoprotein(HDL),early renal disfunction,3-vessel lesions and left main lesion in coronary arteries.Conclusions It is useful to perform selective renal artery angiography following coronary angiography.ARAS is not parallel with CHD.Female,hypercholesterolemia,low HDL,early renal disfunction,CHD with 3-vessel lesions and left main lesion are the independently predicts risks for ARAS.

Abstract:Aim To study the relationship between type of hypertension and multiple metabolic disorder in rural non-treated hypertensive individuals in Fuxin of Liaoning Province.Methods Multistage sampling of a resident(≥5 years) group with non-treated hypertension over 35 year were clustered and divided into 3 groups by type of hypertension.The relationship between type of hypertension and multiple metabolic disorders was analyzed by SPSS11.5 statistical software.Results 4273 non-treated hypertensive individuals were contained in this study aged 56.5±11.4.In the isolated systolic hypertension(ISH) group,the prevalence of overweight/obesity(OW/OB),impaired glucose metabolism disorder,and dyslipidemia was 31.4%,19.9%,and 46.9%,respectively.In the isolated diastolic hypertension(IDH) group,the prevalence of OW/OB,impaired glucose metabolism disorder,and dyslipidemia was 41.9%,11.7%,and 44.3%,respectively.In the systolic-diastolic hypertension group,the prevalence of OW/OB,impaired glucose metabolism disorder,and dyslipidemia was 45.6%,19.0%,and 53.3%,respectively.The prevalence of overweight/obesity(OW/OB) in all age groups was different among 3 groups,but the prevalence of impaired glucose metabolism disorder was different among 3 groups only in the <60 age group.In the isolated systolic hypertension(ISH) group,the prevalence of impaired glucose metabolism and dyslipidemia was lower in middle-age group than that in old-age group;however the prevalence of OW/OB was higher in middle-age group.In other groups only the prevalence of OW/OB was higher in middle-age group than that in old-age group.In the third group,there was higher prevalence of multiple metabolic disorders,except few.Conclusion The prevalence of multiple metabolic disorders between different types of hypertension was different.We should take different controlling measures to prevent cardiovascular disease.

Abstract:Aim To develop a new animal model for the investigation of adventitial function in atherosclerosis.Methods One carotid artery adventitia of rabbit with 2 mg/mL typeII collagenase was digested,then adventitia with microforceps was removed,and the contralateral carotid artery was used as control.HE staining and immunohistochemistry were performed to assure adventitia was removed,and to assess intimal change after adventitial removal.Results It is feasible to remove vascular adventitia with collagenase digestion and mechanical dissection.Removal of adventitia of rabbit carotid artery induced neointima development in 2 weeks.Cells in neointima were secreating smooth muscle cells.The other sides were normal.Conclusions The method we used to remove artery adventitia is effective.Normal adventitia is necessary for normal vessel structure.