Abstract:Aim To investigate the effects of ganoderma lucidum polysaccharide on endothelial function of type 2 diabetic rats in vivo,and explore the potential mechanisms of ganoderma lucidum polysaccharide on protecting the endodermis of blood vessel. Methods SD rats were fed with high fat diet for 4 weeks,then injected STZ(30 mg/kg) to replicate type 2 diabetic rats.The successful diabetic rats model were randomized into normal control group,diabetes group,berberine(30 mg/kg) in the positive control group,ganoderma lucidum polysaccharide of low,middle and high-dose group(Ganoderma lucidum polysaccharide were orally given 200 mg/kg,400 mg/kg and 800 mg/kg).After 16 weeks' treatment,the thoracic aorta was immediately isolated.Endothelium-dependent relaxation induced by acetylcholine,the activity of catalase(CAT),glutathione peroxidase(GSH-PX),total cholesterol(TC) and triglycerides(TG) in serum were detected. Results After treatment the blood glucose concentration of middle-dose group and high-dose group was lower than diabetes group(P<0.01).The advanced glycation end product(AGE) in serum was significantly lower in high-dose group than diabetes group(P<0.01),catalase and glutathione peroxidase in serum was significantly higher in high-dose group than diabetes group(P<0.01),TC and TG in serum was significantly lower in high-dose group than diabetes group(P<0.01). Conclusion Ganoderma lucidum polysaccharide is able to protect against aorta endothelial dysfunction in diabetic rats.The mechanisms may be involved in decreaseing advanced glycation end products in serum,enhancing the activity of CAT and GSH-PX in serum and accommodating dyslipidemia.

Abstract:Aim To investigate the protective effect of acetaminophen on the injury of human umbilical vein endothelial cells(HUVEC) induced by hydrogen peroxide in vitro and to explore its underlying mechanism. Methods The HUVEC line was subcultured in vitro and used for experiment.This study was conducted as follows: normal control group,hydrogen peroxide(H2O2)-injury group(0.1 mmol/L),acetaminophen protective group(25 μmol/L,50 μmol/L and 100 μmol/L) was pretreatmented for 1 hour.The cell viability was measured by MTT assay.The content of malondialdehyde(MDA) and super oxide dismutase(SOD) was determined by kit.The expression of apoptosis-related protein caspase-3 was determined by Western blotting.Apoptosis of the cells was analyzed by flow cytometry. Results Comparied with normal control group,H2O2 can obviously damage vascular endothelial cells(P<0.05).It shows as the decrease of viability of HUVEC,the increase of the content of MDA,the decrease of SOD activity,the increase of caspase-3 expression and apoptosis rate.Acetaminophen can disturb the damage of H2O2-induced endothlial celles.It can increase viability of HUVEC,decrease the content of MDA,increase SOD activity,decrease caspase-3 expression and apoptosis rate.The effect of acetaminophen showed a dose-dependent manner(P<0.05). Conclusions The results show that acetaminophen can decrease the damage of cultured vascular endothelial celles injured by H2O2.It also can inhibit the apoptosis of vascular endothelial cells induced by H2O2.The protective effect of acetaminophen may relate with antioxidation and inhibiting pro-apoptotic protein caspase-3.

Abstract:Aim To investigate the dynamic changes of the expression of HCN2 and HCN4 mRNA and protein in infarcted border zones of post-AMI heart. Methods AMI model induced by the left anterior descending coronary artery(LAD) ligation in Sprague-Dawley rats,were randomly divided into 24 hours group,1-week group,2-week group,4-week group.Accordingly,the sham-operation group was established.Left ventricular infarcted border zone myocardial tissues were harvested at each time,meanwhile,corresponding myocardial tissues in sham-operated rats were harvested too.HCN2 and HCN4 mRNA and protein expression were measured by RT-PCR,immunohistochemistry and Western blotting. Results The expression of HCN2 and HCN4 mRNA and protein were detected in sham-operated rats.The expression of HCN2 and HCN4 mRNA and protein underwent a dynamic change process.The expression of two of them exhibited an ascending tendency at 24 hours after AMI,reached peak value at 1 week after AMI,decreased gradually after that.But the expression of HCN2 channel was also increased compared with sham-operated group at 4 week after AMI,while there was no significant difference in HCN4. Conclusion Expression of HCN2 and HCN4 mRNA and protein in infarcted border zone myocardial tissues underwent a dynamic change process and reached peak value at 1 week after AMI.

Abstract:Aim To study the effect of oxidized low density lipoprotein on biological properties of smooth muscle progenitor cells. Methods Mononuclear cells were isolated from human peripheral blood and cultured with fibronectin,platelet-derived growth factor-BB and EGM-2 medium. Cellular immumofluorescence method and reverse transcription polymerase chain reaction(RT-PCR) method were used to indentify smooth muslce progenitor cells.Smooth muslce progenitor cells were cultured in medium respectively added with oxidized low density lipoprotein in 25,50,100,200 mg/L for 24 hours.Then MTT assay was used to detect cellular proliferational ability,cellular immumofluorescence method and RT-PCR method were used to detect the expression of α-smooth muscle actin. Results Smooth muscle progenitor cells grew with a hill and valley morphology.Smooth muscle progenitor cells could express smooth muslce cell specific biomakers such as α-smooth muscle actin and calponin.The proliferation ability of experimental groups was significantly higher than that of the control group.Especially when the concentration of oxidized low density lipoprotein was 50 mg/L,the proliferation ability of smooth muscle progenitor cells was the highest.The expression of α-smooth muscle actin in experimental group was less than that in control group. Conclusion Oxidized low density lipoprotein can stimulate proliferation and decrease α-smooth muscle actin expression of smooth muscle progenitor cells.

Abstract:Aim To investigate the inhibition of naftopidil ramification YMⅢ on Wistar rat and spontaneously hypertensive rat(SHR) thoracic aorta smooth muscle cell(tASMC) proliferation induced by angiotensin Ⅱ(AngⅡ),and to explore the possible mechanisms. Methods Primary culture of tASMC from Wistar rat and SHR was established in vitro,and the effects of YMⅢ on tASMC proliferation induced by AngⅡ were determined by methyl thiazolyl tetrazolium(MTT) assay.The effects of YMⅢ on the expressions of angiotensinogen(AGT) and c-myc mRNA were detected by real-time reverse transcription polymerase chain reaction(RT-PCR). Results 0.1 μmol/L YMⅢ depressed Wistar rat and SHR tASMC proiferation and YMⅢ(0.01,0.05,0.1 μmol/L) depressed tASMC proiferation induced by AngⅡ in a concentration-dependent manner.YMⅢ(0.05～0.1 μmol/L) down-regulated the expression of AGT and c-myc mRNA on Wistar rat in 24 hours.YMⅢ(0.01,0.05,0.1 μmol/L) down-regulated the expression of AGT and c-myc mRNA on SHR in 24 hours. Conclusions Proliferation of tASMC induced by AngⅡ can be inhibited clearly by YMⅢ,the effect is more significant on SHR than on Wistar rat.The mechanisms may be related to down-regulating the expression of AGT and c-myc mRNA.

Abstract:Aim To explore the effect of melatonin(MT) on the proliferation and apoptosis in endothelial progenitor cells(EPC) from patients with coronary artery disease in vitro. Methods Total mononuclear cells were isolated from patients with coronary artery disease in vitro by Ficoll density gradient centrifugation,then the cells were plated on fibronectin-coated culture dishes.After 7 days,the attached cells were treated by MT with different concentrations(0,0.5,1.0,2.0 mmol/L) for different hours(6,12,24,48 h).EPC were identified by demonstrating the expression of CD34,VEGFR-2 and CD133 under a laser scanning confocal microscope.MTT assay was used to detect the effect of MT on the proliferation ability of EPC.EPC adhesion assay was performed by replating it on fibronectin-coated dishes,and the adherent cells were then counted.EPC migration were assayed by modified Boyden chamber assay.Flow cytometry was used to detect cell apoptosis.The expressions of bcl-2 mRNA and protein were detected respectively by RT-PCR and western blot. Results After exposured to MT,the proliferation,adhesion and migration of EPC in MT group was higher,and the cell apoptosis rate was lower than that in control group in a dose-dependent and time-dependent manner(P<0.01).Expression of bcl-2 mRNA and protein of EPC in MT group was higher than that in control group(P<0.01). Conclusion MT promotes the proliferation,adhesion and migration of EPC and inhibits the cell apoptosis via upregulating bcl-2 expression.

Abstract:Aim To explore the influence of advanced glycation end products(AGE) on inducing rat retinal microvascular endothelial cells(RMEC) to form new blood vessels by setting up a stable angiogenesis culture system on Matrigel and further discuss the effect of AGE on diabetic retinopathy. Methods RMEC were cultured in vitro and AGE-BSA were prepared.A stable angiogenesis culture system was set up on Matrigel.The experiment was divided into different concentration and different time groups.The expression of CD34 was detected by immunocytochemical method.The results were tested by calculating the number of tubules in microscopic observation and computer image-analysis.Finally the data was analyzed by statistics. Results RMEC were lumina formation on Matrigel.After treatment with AGE-BSA,the number of tubules increased and in certain ranges showing a dose-dependent and time-dependent manner(P<0.01). Conclusion AGE can promote inducing RMEC on Matrigel in vitro.

Abstract:Aim To investigate the effects of scavenger receptor class B type Ⅰ(SR-BⅠ) in atherosclerosis by observing the change of SR-BⅠ expression. Methods Eighty patients with hypercholesterolemia and sixty healthy persons were investigated as hyperlipidemia group and control group respectively.Blood samples were extracted from ulnar vein.Blood serum,plasma and platelets were extracted and stored at a refrigerator of-80℃.The level of angiotensin Ⅱ(AngⅡ) in plasma and nitric oxide(NO) in serum were detected by the method of radioimmunity and nitrate reductase respectively.The expression of SR-BⅠ mRNA and protein on platelet were assessed by real-time polymerase chain reaction(RT-PCR) and Western Blotting respectively. Results The plasma AngⅡ and serum NO of hyperlipidemia group were higher than that of control group(P<0.01).The expression of SR-BⅠ mRNA and protein on platelets were lower in hyperlipidemia group than those of control group(P<0.05 and 0.01).The expression of SR-BⅠ protein was inversely correlated with the plasma AngⅡ(r=-0.488,P<0.05) in patients with hypercholesterolemia. Conclusion The expression of SR-BⅠ decreases and the decrease of SR-BⅠis related to AngⅡ.

Abstract:Aim To explore the relationship of common carotid intima-media thickness(CIMT)and the coronary lesions' morphology with the thrombolysis in myocardial infarction(TIMI) risk score for non-ST-elevation acute coronary syndrome(NSTEACS). And evaluate the roles of CIMT in risk stratification of NSTEACS. Methods One hundred and thirty-two patients with NSTEACS were recruited.CIMT were measured,and the coronary angiographies were analysed to detect the single or multiple complex coronary stenostic lesions.Their correlation with TIMI risk score and its variables were investigated. Results Satisfying images of CIMT were obtained in one hundred and twenty-three patients(99.2%),and the general CIMT was 0.83±0.22 mm.Sixty-two patients(50.4%) had an abnormal(≥0.8 mm) CIMT,whilst 52 patients(39.4%) only had single complex coronary lesions and 80(60.6%) had multiple complex coronary lesions.In patients with abnormal CIMT,more of them were ≥65 years old,had diabetes,or with previous ischemic heart disease,than in those without abnormal CIMT;In patients with multiple complex coronary lesions,more of them were ≥65 years old,had diabetes,or with previous ischemic heart disease,than in those with single complex coronary lesions.CIMT was correlated with TIMI risk score(r=0.25,P=0.004),whilst the presence with multiple complex lesions was associated with TIMI risk scale(r=0.31,P<0.01).Using a Logistic regression analysis,the presence of an abnormal CIMT was only related to age ≥65 [OR: 3.52(CI: 1.48～9.37),P=0.001] and diabetes mellitus [OR: 3.83(CI: 1.66～8.91),P=0.004].The presence with multiple complex lesions was also associated with age ≥65 [OR: 17.32(CI: 6.53～52.34),P<0.001] and diabetes mellitus[OR: 3.06(CI: 1.84～8.73),P=0.006]. Conclusion CIMT and the presence of multiple complex lesions in patients with NSTEACS are correlated with TIMI risk score.Both variables were related to age and diabetes.CIMT can play a role in the risk stratification of NSTEACS.

Abstract:Aim To evaluate coronary artery calcification(CAC) by multislice spiral CT scan in patients starting to undergo hemodialysis,and to analyse its association with cardiovascular events in the follow-up. Methods 29 patients starting to undergo chronic hemodialysis(duration of hemodialysis under 6 months) were enrolled in this study.CAC score,along with related parameters,was quantified by Multislice Spiral CT scan at the initiation of this study.The patients were followed up for about 18 months for appraising cardiovascular events. Results 18 cases of CAC(62.07%) were identified by CT scan in 29 patients starting to undergo chronic hemodialysis,with average CAC score being 482.41±739.97.During the 18 months follow-up,significant difference of cardiovascular events was documented among patients with CAC(22 times) and without CAC(1 time)(P=0.001).CAC was independently associated with cardiovascular events(Spearman's Rho=0.882,P<0.0005).Stepwise regression analysis in 29 patients indicated that CAC score was related with age and serum PO4(P=0.013,P=0.043).However,CRP,Ca,iPTH,Chol,TG,HDL-C,LDL-C,BMI,SBP and DBP were excluded from this regression equation(P>0.05). 6 cases(15.79%) were died of cardiovascular events during the 18 months follow-up,including 5 cases with CAC and 1 case without CAC.Kaplan-Meier survival analysis showed that there was no significant difference between patients with CAC and without CAC(P=0.2930). Conclusion Multislice Spiral CT based coronary artery calcification scoring is helpful in risk evaluation of cardiovascular events of hemodialysis patients.

Abstract:Aim To investigate the correlation between severity of coronary artery lesions and middle-term prognosis with obstructive sleep apnea-hypopnea syndrome(OSAHS) in patients with acute coronary syndrome(ACS). Methods The patients with ACS were classified into four groups according to results of coronary angiography and polysomnography: ACS group(group Ⅰ),ACS complicated with mild OSAHS(group Ⅱ),ACS complicated with moderate OSAHS(group Ⅲ),ACS complicated with severe OSAHS(group Ⅳ).Coronary artery lesions' severity was compared by Gensini score.The relationship between Gensini score and apnea hypopnea index(AHI) and nocturunal lowest blood oxygen saturation(LSaO2) were analyzed.The patients were followed up for six months,and the amounts of cardiovascular composite endpoint in four groups were compared. Results Compared with group Ⅰ,AHI of group Ⅱ,Ⅲ and Ⅳ were much higher(P<0.01),LSaO2 of group Ⅱ,Ⅲ and Ⅳ were much lower(P<0.01).Compared with group Ⅱ,AHI of group Ⅲ and Ⅳ were much higher(P<0.01),LSaO2 of group Ⅲ and Ⅳ were much lower(P<0.01).Compared with group Ⅱ and Ⅰ,Gensini scores of group Ⅲ and Ⅳ were much higher(P<0.01).Gensini score of group Ⅳ was higher than group Ⅲ(P<0.05).Gensini score was positively correlated with AHI and negatively with LSaO2.Incidence of cardiovascular composite endpoint events during six months in four groups were 19.35%,22.58%,32.26%,43.33% respectively.The group Ⅳ's cardiovascular composite endpoint events was higher than group Ⅰ(P<0.05).Total cholesterol,triglyceride,low density lipoprotein cholesterol and classification of OSAHS were independent predictors for cardiovascular composite endpoint events. Conclusion Moderate and severe OSAHS is related to the severity of coronary artery lesions in patients with ACS.Complicated OSAHS in patients with ACS may increase the incidence rate of cardiovascular composite endpoint events,especially severe OSAHS.OSAHS is an independent predictor for cardiovascular composite endpoint events in patients with ACS.

Abstract:Aim To study the relationship between serum uric acid(SUA) and coronary artery disease(CAD),and try to explain the relationship between them by observing the effect of uric acid on vascular smooth muscle cells(VSMCs) of SD rats. Methods The basic informations(fast blood fat,glucose,uric acid and body mass index,et al) of 368 in-patients undergone coronary artery angiography were collected.Uric acid levels were compared among patients with different degree of coranary lensions,and Logistic regression was applied to analyze the relationship between serum acid level and degree of coronary lensions.Vascular smooth muscle cells were cultured with attachment blook method.Different concentrations of uric acid were applied to culture together with them for 48 hours,and the vital force of VSMCs was detected and compared. Results Levels of SUA were higher in male patients and in patients with CAD,and were relatively higher in patients with three coronary arteries lesions and in patients with coronary lesion score over 15.Taking coronary lesion score as dependent variable,SUA level didn't enter regression equation.Effects of different concentrations of uric acid on VSMCs weren't statisticly different. Conclusions Serum uric acid isn't an independent risk factor of CAD.Uric acid have no definite effects on VSMCs of rats.

Abstract:Aim To determine the effect of atorvastatin on macroangiopathy in patients with type 2 diabetes mellitus(DM) and further to explore the possible mechanism. Methods 120 patients with DM were enrolled in the present study and randomly divided into routine treatment group and atorvastatin treatment group.Flow-mediated dilation(FMD) and intima-medial thickness(IMT) was measured before treatment and after 6-month treatment.The levels of plasma monocyte chemoattractant protein-1(MCP-1),nitric oxide(NO),and vonwillebrand factor(vWF) were also determined. Results After 6 months therapy,the levels of blood glucose and blood pressure were well controlled in two groups.FMD and the levels of NO and vWF were also improved in both groups.However,compared with routine treatment group,endothelial function was more significantly improved by treatment with atorvastatin(P<0.05).Atorvastatin treatment controlled blood lipid,attenuated the levels of MCP-1 and the progress of IMT(P<0.05),but routine treatment had no effect. Conclusion Atorvastatin may induce a markedly protective effect on the macroangiopathy of DM,which may be related to regulating blood lipids,protecting vascular endothelium and inhibiting vascular inflammation.

Abstract:Aim To study the apolipoprotein E(apo E) gene polymorphism relating to the severity of coronary artery disease(CAD) and its impact on prognosis. Methods Polymerase chain reaction-restriction fragment length(PCR-RFLP) was used to determine apoE genotype of 251 CAD patients and 200 controls.Plasma lipid levels were measured by routine ways. Results Five kinds of apoE genotype,ε3/3,ε3/2,ε4/3,ε4/2 and ε4/4 were detected in the study.It indicates that different kind of apo E allele had different function on plasma lipid and lipoprotein levels.There were statistic differences in apoE ε4/3 genotype and ε4 allele frequencies between normal-lipid-leveled CAD patients and controls,but between the two hyperlipidemia subgroups,no differences had been found.Patients with heart function of class Ⅲ~Ⅳ had higher apoE ε4/3 genotype and ε4 allele frequencis and lower ε3/3 genotype frequencies than those with better heart function(class Ⅰ~Ⅱ). Conclusion Apo E gene polymorphism is involved in the occurrence,the development and the heart of CAD.ApoE ε4/3 genotype and ε4 allele are important genetic fators of CAD and may have relationship with worse heart function or cardiac events,while ε3/3 genotype might be a protective factor of CAD.

Abstract:Aim To investigate the clinical features and therapeutic efficacy of thrombolytic therapy of acute myocardial infarction(AMI) in young patients and older patients. Methods 24 patients ≤45 years of age with a diagnosis of AMI,were received thrombolytic therapy after the onset of AMI,and assigned to the young group.40 consecutive patients aged 50～70 years old were selected as the control group.The patient's cardiovascular history,risk factors(smoking,hypercholesterolemia,hypertension,diabetes mellitus,and family history) and therapeutic efficacy of thrombolytic therapy were recorded and analysed. Results The percentage of male in young group were higher than in older group(P<0.01).The percentage of smoking and family history in young group were higher than in older group(79.2% and 45.8% vs 40.0% and 12.5%,P<0.01).The percentage of hypertension and diabetes mellitus in older group were higher than in young group(60.0% and 40.0% vs 16.7% and 12.5%,P<0.05).The history of angina pectoris in young group were shorter than in older group(324±576 days vs 928±1 846 days,P<0.001).The patients of chest pain in 30 days and 12 hours in young group were much more than in older group(62.5% and 41.7% vs 10.0% and 5.0%,P<0.001).The initiating time of reperfusion had no significant difference in two groups,but the recanalization rate of coronary artery was higher in young group than in older group(75.0% vs 50.0%,P<0.05). Conclusion Smoking and family history were the most common risk factors among young patients.The clinical history of young patients was shorter than older patients.The therapeutic efficacy of thrombolytic therapy in young patients was more excellent than in older patients.

Abstract:Aim To investigate the clinical application of ankle-brachial index(ABI) in the treatment of peripheral arterial disease in the patients with type 2 diabetes mellitus. Methods 112 type 2 diabetic patients with peripheral arterial disease were randomly divided into drug treatment group(n=62) and interventional therapy and drug treatment group(n=50).All patients were checked for ABI by automatic analysis system of angiosclerosis. Results ABI was less than 0.90 in type 2 diabetic patients with peripheral arterial disease before therapy.ABI was increased in type 2 diabetic patients with peripheral arterial disease after interventional therapy and drug treatment(P<0.05).ABI was not increased in type 2 diabetic patients with peripheral arterial disease after drug treatment(P>0.05). Conclusion ABI is significant in type 2 diabetic patients with peripheral arterial disease who had taken the interventional therapy.

Abstract:Aim To investigate the relation between Lp-PLA2 levels and degree of carotid atherosclerosis,and meanwhile to explore the relation between lipids、hs-CRP and Lp-PLA2 levels. Methods 138 cases(70 male cases and 68 female cases) were collected and their bilateral carotid arteries were examined by color Doppler ultrasound.According to the results of color Doppler ultrasound,the 138 cases were divided to four groups:30 cases with normal carotid arteries;38 cases with intima-media incrassation of carotid arteries;and 70 cases with plaque.The Lp-PLA2 concentration of limosis blood serum was determined by ELISA method.hs-CRP concentration and total cholesterol(TC),triglycerides(TG),low density lipoprotein(LDL) were determined as well. Results The concentration of TC,hs-CRP,and Lp-PLA2 were lower in the group with normal carotid arteries and intima-media incrassation of carotid arteries than those in the groups with plaque,and there were significant differences(P<0.05).No significant difference was found in TG and LDL among the three groups(P>0.05). The concentration of Lp-PLA2 and TC,LDL,hs-CRP are of significantly positive correlation(r=0.279,0.187,0.176,P=0.001,0.028,0.039).No significant difference was found between Lp-PLA2 and TG(r=0.100,P=0.244). Conclusion The Lp-PLA2 was associated with CAS degree as a factor of promoting atherosclerosis,especially of the negativity or positivity of the exisitence of plaque.The mechanism may be related to lipids and inflammation.

Abstract:Aim To discuss the importance of insulin,high-sensitivity C-reactive protein(hs-CRP) and adiponectin in acute cerebral infarction(ACI). Methods The differences and relationship of insulin,hs-CRP and adiponectin were discussed between 45 patients with ACI and 36 patients of control group. Results The expressions of hs-CRP and insulin were higher in ACI group than in control group.However,the expression of adiponectin was lower in ACI than in control group. Conclusion The increase of insulin,hs-CRP and the decrease of adiponectin exist in ACI,the expression of hs-CRP was relative with adiponectin,insulin.