Abstract:Aim To investigate the changes of myocardial apoptosis and peroxisome proliferator activated receptor α of aging rats and the effects of atorvastatin on it.Methods 30 wistar rats at 20 month old were divided into three groups:aging control group,high dose atorvastatin group [10 mg/(kg·d)],low dose atorvastatin group [1 mg/(kg·d)] and another 10 wister rats at 3 month old as young control group.Atorvastatin was delivered by intragastric administration for 4 month.Normal sodium was delivered to control group's rats by intragastric administration too.Cardiac apoptosis was evaluated by terminal deoxynucleotidy transferase mediated dUTP-biotin nick end labelling(TUNEL)assay.Peroxisome proliferator activated receptor α protein was detected by western blot.Results ①The expression level of peroxisome proliferator activated receptor α protein in aging control group is significantly lower than that of young control group(P<0.01),atorvastatin upregulate peroxisome proliferator activated receptor α expression obviously(P<0.01);②The density of TUNEL-positive cells in aging group was significantly higher than that of young control group(P<0.01).Compared with aging group,the density of TUNEL-positive cells decreased markedly in both atorvastatin group.Conclusions Atorvastatin inhibits the myocardic apoptosis of aging rats,which may be related with its upregulating the expression level of peroxisome proliferator activated receptor α.

Abstract:Aim To investigate the effects of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)on pregnancy-associated plasma protein-A(PAPP-A)expression in endothelial cells and analyse its mechanism.Methods The rat aortic endothelial cells were obtained from thoracic aortas and cultured by tissue explant method.Passage 3～4 cells were used in the experiment.Rortic endothelial cells were coincubated with different concentrations of TNF-α(10,20,40,60 and 100 μg/L)and IL-1β(1,5,10,20 and 50 μg/L)for 24 h or coincubated with 60 μg/L TNF-α,20 μg/L IL-1β for different periods(2,4,8,16,24 and 48 h).BAY11-7082(20 μmol/L)was pretreated for 60 min before rortic endothelial cells were coincubated with 60 μg/L TNF-α,20 μg/L IL-1β for 48 h.LDH activity in supernatant was detected.PAPP-A mRNA and protein expression were detected by RT-PCR or ELISA.Results After the intervention of TNF-α or IL-1β for 24 h,the levels of PAPP-A expression increased with the concentration raise(P<0.05);Cytokines stimulated PAPP-A expression in the two groups(P<0.05);BAY11-7082 inhibited PAPP-A expression stimulated by TNF-α and IL-1β.Conclusions TNF-α and IL-1β may stimulate the expression of PAPP-A,nuclear factor-κB(NF-κB)activation may be the primary mediator of cytokines stimulated PAPP-A expression.

Abstract:Aim To explore the role of heat-shock protein 90 in protective effect of hydrogen sulfide against H9C2 cardiac cells injuries induced by cobalt chloride.Methods H9C2 cells were exposed to cobalt chloride at different doses to set up the chemical hypoxia-induced cardiomyocyte injury model.Sodium hydrosulfide(a hydrogen sulfide donor)was added into cell medium for 30 min before cobalt chloride treatment.Cell viability was tested by using cell counter kit-8.Morphological changes in apoptotic cardiomyocytes were detected by Hoechst 33258 staining and photofluorography.The expression of heat-shock protein 90 was evaluated by Western blot.Results H9C2 cell viability was inhibited by cobalt chloride at the concentrations from 600 to 1000 μmol/L for 24 h in a dose-dependent manner.Pretreatment with 400 μmol/L sodium hydrosulfide 30 min before exposure to cobalt chloride significantly blocked the cardiomyocyte cell injuries induced by cobalt chloride at 600,800 and 1000 μmol/L respectively,leading to an increase in cell viability.Heat-shock protein 90 expression was upregulated after treatment with 400 μmol/L sodium hydrosulfide for 30 min,peaking at 6 h to 9 h,returning to the basal level at 18 h.17-allylamino-17-demethoxygeldanamycin(2 μmol/L),an inhibitor of heat-shock protein 90,not only enhanced H9C2 cells injury induced by cobalt chloride,but also obviously blocked the inhibition of hydrogen sulfide on cobalt chloride-induced cardiomyocyte damage,reducing viability of H9C2 cells,increasing number of apoptotic cells,which didn't damage H9C2 cells alone.Conclusion Hydrogen sulfide can protect H9C2 cells against cobalt chloride-induced injury and upregulate expression of heat-shock protein 90.17-allylamino-17-demethoxygeldanamycin not only increases cobalt chloride-induced H9C2 cell injury,but also significantly inhibits the cardioprotection of hydrogen sulfide,suggesting that heat-shock protein 90 may mediate the cardioprotective effect afforded by hydrogen sulfide.

Abstract:Aim To investigate the effect of arecoline on inflammation factors expression of RAW 264.7 murine macrophages and the relation with PPARγ.RAW 264.7 murine macrophages were co-incubated with 75 mg/L oxidized low density lipoprotein and arecoline respectively for 24 h.Methods Reverse transcription-PCR was performed to determine the lever of the inflammation factor and PPARγ mRNA expression.Results The cell viability was decreased when culture with the arecoline 10-3 mol/L,but not in the group of arecoline 10-6 mol/L,10-5 mol/L and 10-4 mol/L.The inflammation factor expression in macrophages has no significant difference among control group,10-6 mol/L,10-5 mol/L and 10-4 mol/L arecoline groups.The mRNA levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and intercellular adhesion molecules-1(ICAM-1)was no significant changes when the macrophages was co-incubated with 75 mg/L ox-LDL and 10-6 mol/L arecoline,but they were all decreased in ox-LDL 75 mg/L+10-4 mol/L arecoline group and ox-LDL 75 mg/L+10-5 mol/L arecoline group while the expression of PPARγ mRNA was increased in ox-LDL 75 mg/L+10-5 mol/L arecoline group.Conclusion These results suggest that arecoline could inhibit inflammation factor expression of macrophages,which may be related to up-regulation of PPARγ.

Abstract:Aim To package heat shock protein 72 recombinant adenoviruses and to explore its effect on ECV304 cell apoptosis.Methods Heat shock protein 72 recombinant adenoviruses was generated by employing homologous recombination in bacteria,flow cytometric analysis was used to invest the effect of heat shock protein 72 recombinant adenoviruses on cell apoptosis,luciferase reporter system and western blotting were used to analyse its effects on p53 Transcription and expression.Results Heat shock protein 72 recombinant adenoviruses were successfully packaged,and heat shock protein 72 could enhance cell apoptosis.Conclusions Heat shock protein 72 may contribute to cell apoptosis by upregulating P53 expression.

Abstract:Aim To prokaryotic express mouse obese gene in E.coli expression system and purify the fused mouse leptin.Methods Mouse obesity gene leptin was amplified from pMET mouse leptin and subcloned in pET-28a(+)to construct recombinant vector pET-28a-lep.The sequence of pET-28a-lep was checked by sequencing and restriction analysis.Expression of BL21-pET-lep transformed with pET-28a-lep was induced with 0.1 mmol/L isopropylthiogalactoside and analyzed with Sodium dodecylsulfonate-polyacrylamide gel electrophoresis and Western Blotting to identify the mouse leptin molecular weight and optimum time for purpose protein expression.The maximum amount of the fused protein expressed was examined with thin layer chromatography.Mouse leptin dissolved with 0.5% Sarkosyl was purified by Ni2+ affinity chromatography column.Results Sequencing and restriction analysis confirmed the right sequence of pET-28a-lep.Sodium dodecylsulfonate-polyacrylamide gel electrophoresis and Western Blotting indicated a 22.5 kDa fused mouse leptin,in the form of inclusion body,was expressed with high efficiency in BL21-pET-lep.The maximum amount of the fused protein produced was 39.2% of the total cellular protein.The concentration of purified protein is 1.086 g/L.Conclusion BL21-pET-lep could highly express recombinant mouse leptin.This provides a basis for further researches on biological activity of mouse leptin and its role on the developing process of arthrosclerosis.

Abstract:Aim To study the effects of inhibition of NF-κB in the early phase after myocardial ischemia on myocardial reparation in rats.Methods The myocardial ischemic injury models were established by hypodermic injection of high dose of ISO in rats.Surviving rats were randomly divided into experimental group and treated group,meanwhile control group was set.Then treated group was injected PDTC 100 mg/(kg·d)for 26 days.Myocardial pathomorphologic changes,NF-κBp65 activation,capillary density and myofibroblast accumulation were investigated.Then cardiac structure and function were assessed by echocardiography on day 14 and 28 of experiment.Results Cardiac necrotic areas were similar between experimental group and treated group.PDTC inhibited the activation of myocardial NF-κB,delayed the replacement of the damaged myocardium with granulation tissue,significantly depressed capillary density in the border area of necrosis and decreased myofibroblast accumulation in the zone of necrosis.The ratio of ventricular aneurysm formation in treated group was higher than experimental group.On day 28,the left ventricular functions in treated group were worse than that in experimental group.Conclusion Inhibition of NF-κB in the early phase after experimental myocardial ischemic injury in rats may be adverse to myocardial reparation,and may aggravate heart failure.

Abstract:Aim To investigate the expression and significance of interleukin-18 and matrixmetalloproteinase-9 in human coronary arteries with different atherosclerotic lesions.Methods 48 coronary arteries specimens from 14 autopsy cases were divided into 3 different groups according to HE staining,which were unstable plague group,stable plague group and normal group.These specimens were detected by immunohistochemistry and reverse transcription-polymerase chain reaction for protein and mRNA analysis.Results Interleukin-18 and matrixmetalloproteinase-9 mRNA and protein expression can be detected in atherosclerosis,and there is significant difference between unstable plaques and stable plaques.The levels are significantly higher in unstable plaques.Positive association between interleukin-18 and matrixmetalloproteinase-9 expression intensity showed in Spearman tests.Conclusions These results indicates interleukin-18 and matrixmetalloproteinase-9 both participate in the initiation and development of atherosclerosis and are highly relevant to the unstable plaque.These findings suggests that interleukin-18 is in coordination with matrixmetalloproteinase-9 during the development of atherosclerosis.

Abstract:Aim To study the effect of advanced glycation end products(AGE)on the expression of extracellular matrix metalloproteinase inducer(EMMPRIN)and the activity of matrix metalloproteinase-9(MMP-9)in cultured mouse macrophage(J774A.1).Methods The AGE-BSA was prepared by incubating bovine serum albumin(BSA)with glucose.The cultured J774A.1 was intervented with AGE-BSA(50,100,200,400 mg/L)for 24 h or 200 mg/L AGE-BSA for 12,24,48 h,respectively,taking DMEM and BSA as negative control.The mRNA expression of EMMPRIN in J774A.1 was analyzed by reverse transcription-polymerase chain reaction(RT-PCR)and the concentration of EMMPRIN in the supernatant was quantified by enzyme linked immunosorbent assay(ELISA).The activity of MMP-9 in J774A.1 was determined by gelatin enzymogram method.Results Compared with that in the cells incubated with DMEM and BSA,the level of EMMPRIN mRNA,the concentration of EMMPRIN in the supernatant and the activity of MMP-9 in the cells incubated with 50,100,200 or 400 mg/L AGE-BSA,respectively,for 24 h or with 200 mg/L AGE-BSA for 12,24,48 h,respectively,was increased in a dose and time dependent manner significantly(P<0.05).Moreover,with the increasing concentration of AGE-BSA or the extended incubating time,the level of EMMPRIN mRNA,the concentration of EMMPRIN in the supernatant and the activity of MMP-9 increased singnificantly(P<0.05).Concluions The AGEBSA can stimulate the expression of EMMPRIN,the secretion of EMMPRIN and the activity of MMP-9 in cultured J774A.1,indicating its effect on atherogenesis and plaque rupture.

Abstract:Aim To study arteriosclerosis induced by large dose of vitamin D3(VitD3)and its mechanism in mouse.Methods Forty Kunming mouse were randomly divided into three groups including control group,model group and normal VitD3 group,with sixteen in the forner two groups,in which male and female were eight equally and eight in the last group.Then mouse in model group were given forty units of VitD3 for three days and observed for forty-two days.Sections were made for aortic arch and stained by hematoxylin and eosin,thoracic aortawere stained by Von kossa and the percentage of calcificational areas was counted,the content of Ca and P was measured by ICP.Results The media of aorta was seriously calcified in VitD3 model group compared with control group,phenotype of smooth muscle cell in media of aorta have changed to another phenotype which was suspected for chondrocyte,percentage of calcificational areas in model group was remarkably higher than in control group(P<0.01).The content of Ca and P for abdominal aorta was higher in model group than in control group(P<0.05).Conclusion Large dose of VitD3 could lead to the serious calcification located in media of aorta in mouse,and the vascular smooth muscle cell coverted to analogical chondrocyte.The mechanism was connected to that large dose of VitD3 which induces the convertion of vascular smooth muscle cell to analogical chondrocyte and simultaneously leads to the massive deposition of Ca and P in blood vessel.

Abstract:Aim To investigate the relationgship between serum advanced oxidation protein products(AOPP)and atherosclerosis in patients with type 2 diabetes mellitus(T2DM).Methods 90 T2DM patients and 60 healthy controls were enrolled.Serum levels of stromal cell-derived factor-1α(SDF-1α)and AOPP were measured by ELISA and spectrophotometry method separatly.Carotid,femoral and common iliac artery intima-media thickness(IMT)were determined with non-invasive high-resolution B-mode ultrasonography.Results Both serum AOPP and SDF-1α levels were significantly higher in T2DM group as compared with those of heathy controls(80.32±12.65 μmol/L vs 41.80±17.09 μmol/L,P<0.01;2652.05±362.07 μg/L vs 2160.37±424.79 μg/L,P<0.05).Using the mean+2SD of AOPP concentration as cut-off ponit,T2DM with AOPP more than or equal to 92.97 μmol/L were defined as high AOPP group,while less than 92.97 μmol/L as normal AOPP group.Of all the patients with T2DM higher serum SDF-1α levels were found in high AOPP group than those of normal AOPP group(2813.04±330.25 μmol/L vs 2501.73±383.02 μmol/L,P<0.05).Both SDF-1α and AOPP levels were significantly increased in the atherosclerosis(As)group than those of non-As group(2843.93±355.28 μg/L vs 2507.88±360.62 μg/L,P<0.05;89.69±12.58 μmol/L vs 75.10±13.09 μmol/L,P<0.01)in the patients with T2DM.As was defined that carotid or femoral or common iliac artery IMT was more than 1 mm or there was a plaque on the vessal wall according to the ultrasonography.Serum levels of AOPP were correlated positively with serum SDF-1α(r=0.295,P=0.03).Conclusions Protein oxidation damages were increased in T2DM,increased levels of AOPP contribute to the increased expression of SDF-1α on vascular endothelial cells,increased AOPP may be associated with accelerated atherosclerosis in T2DM.

Abstract:Aim To observe the correlation between the level of serum homocysteine and the heart failure in patients with coronary heart diseases and the severity of heart failure.Meanwhile the interventional treatment of folic acid and vitamin B6,B12 were given to the heart failure patients to observe the changes of HCY and the heart failure after the short-term therapy.Methods All 57 patients with chronic heart failure secondary to coronary heart diseases(heart failure group)were divided into three subgroups according to NYHA cardiac function classification(abbreviated as coronary heart function Ⅱ,Ⅲ,Ⅳ groups).Twenty patients who were diagnosed as stable angina pectoris without heart failure were selected as coronary heart disease control group(CHD control group).20 healthy subjects after examination served as healthy control group.The levels of HCY and NT-proBNP were measured.Results ①The prevalence rate of hyperhomocysteinemia(HHCY)was higher in chronic heart failure group than that in CHD control group and the healthy control group.②The serum HCY in patients with chronic heart failure disease were higher than that in stable angina pectoris patients and both higher than that in the healthy people.③ In heart failure group the serum HCY were increased with the NYHA cardiac function classification increase(P<0.05).④ The serum HCY and NT-proBNP was positive linear correlation.⑤In heart failure group,the serum HCY and EF were determined with negative linear correlation;the serum HCY and LVEDD were determined with negative linear correlation.⑥After short-term treated with folic acid,vitaminB6 and B12,the levels of HCY and NT-proBNP were both decreased.Conclusion HHCY are associated with heart failure and maybe one of the independent risk factors of heart failure.The level of HCY is increased with the severity degree of the cardiac function and the serum HCY may participate in the development of heart failure.Both levels of HCY and NT-proBNP seem to be related with the prognosis of heart failure.

Abstract:Aim To compare the early outcomes after coronary artery bypass grafting(CABG)among different aged groups.Method 4 303 patients were underwent isolated CABG in our hospital.All of the patients were divided into 4 groups according to the age:Group A(≤45 years),Group B(46~60 years),Group C(61~74 years)and Group D(≥75 years).The preoperative risk factors,mortality and complications were analyzed between each group.Results The factors that hypertension,the incidence of mild mitral insufficiency or aortic insufficiency,preoperative blood creatinine,the proportion of saphenous vein to left anterior descending artery,ventilation time,intensive care unit stay were lower in group A and B than those in group C and D.Whereas the proportion of cardiopulmonary bypass(CPB)and total arterial bypass grafting were higher in group A and B than those in group C and D.The mortality in group A(0.35%)and B(1.52%)was significant lower than that in group C(2.62%)and D(5.45%).The incidence of complications of atrial fibrillation(8.18%)and low cardiac output syndrome(3.18%)in group D were maximum.Conclusion Saphenous vein to left anterior descending artery,age and CPB are the independent risk factors of operative mortality.The number of diseased vessels,the proportion of total arterial bypass grafting,mitral insufficiency,aortic insufficiency and preoperative blood creatinine are contributory factors to operative death.Postoperative renal insufficiency may be relative to preoperative diabetes mellitus and blood creatinine.

Abstract:Aim To explore the cerebral hemodynamic changes of intracranial artery stenosis in Trancranial Doppler ultrasound(TCD).To compare the sensitivity and specificity of TCD with magnetic resonance angiography(MRA)(or digital subtraction angiography,DSA).Methods 85 cases with cerebral ischemic stroke(CIS)were examined respectively by using TCD and MRA(or DSA).Results Compared with the results of MRA(or DSA),the sensitivity and the specificity of TCD were respectively 88.2% and 98.6% in the diagnosis of intracranial stenosis arteries.Compared with the results of DSA,the sensitivity and the specificity of TCD were respectively 90% and 97.9% in 15 cases with CIS.Conclusion TCD can objectively reflect the cerebral hemodynamic changes in cerebral vessels and can be used to diagnose the stenotic diseases of carotid and intracranial arteries.

Abstract:Aim To observe the effect of Huoxueyihao capsule combined with Levamlodipine on pre-thrombus state of hypertension 2.Methods 152 patients with hypertension 2 were divided into Levamlodipine group,aspirin group and huoxueyihao group,and treated by Levamlodipine Besylate tablet,Levamlodipine Besylate tablet and aspirin enteric-coated tablet,Levamlodipine Besylate tablet and Huoxueyihao capsule for 8 weeks respectively.The levels of blood pressure,nitric oxide(NO),von willebrand factor(vWF),granule membrane protein-140(GMP-140),plasma fibrinogen(Fg)were observed before and after treatment.Results Contrast in the health group,the levels of NO in patients were remarkably decreased(P<0.05),but the levels of vWF,GMP-140 and Fg were markedly increased(P<0.05).The levels of NO in the treated patients were all higher than those of before treatment(P<0.05),and the levels of Huoxueyihao group rise obviously(P<0.05).The levels of vWF,GMP-140,Fg all decreased after treatment(P<0.05),and those in Huoxueyihao group decreased especially(P<0.05).Conclusion Huoxueyihao capsule combined with Levamlodipine could be used in treating pre-thrombus state of hypertension 2 through protecting vascular endothelium and stabilizing platelet function.

Abstract:Aim To evaluate the changes of the arterial function in essential hypertensive(EH)patients with metabolic syndrome(MS).Methods 64 patients with EH were assigned into two groups:EH with MS group(n=32),EH without MS group(n=32).31 healthy subjects were served as normal control.Brachial-ankle pulse wave velocity(BaPWV)was measured using an automatic device(VP1000),arterial elastic index(C1-large artery and C2-small artery)were measured by the CVProfiler-2020 pulse wave analysis instrument.Results BaPWV in EH with MS group(1 916.5±318.0 cm/s)and EH without MS group(1 757.6±313.9 cm/s)were higher than that in control group(1 517.9±315.0 cm/s,P<0.01 and <0.05);and BaPWV in EH with MS group was higher than that in EH without MS group(P<0.05).The magnitude of C1 and C2 reduced in proper order in control group,EH without MS group,EH with MS group(C1:113.2±46.2,91.1±35.0 and 66.6±31.1 mL/mmHg,P<0.01;C2:505±244,355±195 and 253±116 mL/mm Hg,P<0.05).Pearson correlation showed that BaPWV was positively related with age,systolic blood pressure,pulse pressure difference and triglyceride(P<0.01),negatively related with high density lipoprotein cholesterol(HDLC)(P<0.01);C1 and C2 were positively related with HDLC(P<0.05 and 0.01),and negatively related with age,systolic blood pressure,pulse pressure difference and triglyceride(P<0.01).Conclusion MS may aggravate the dysfunction of artery in EH patients.

Abstract:Aim To explore the relationship of serum uric acid(UA),fibrinogen(FIB),low density lipoprotein(LDL)and lipoprotein(a)[Lp(a)] and severity of coronary artery lesions in patients with different types of coronary heart disease(CHD).Methods The investigated population were consisted of 185 cases of CHD patient,including 107 cases of acute coronary syndrome(ACS),30 cases of coronary artery spasm(CAS),48 cases of stable angina pectoris(SAP),and 31 normal controls(NC).Serous UA,LDL and Lp(a)were detected with Hitachi-7600 Auto-analyzer,and FIB was deteced with BE-Thrombolyzer.The severity of coronary artery lesions was quantified with the Gensini score system and the number of impaired arteries.Results Compared with that of NC group,the levels of serous UA were increased in the patients with CAS,SAP and ACS in male;the same phenomenon was observed in SAP and ACS in female.The FIB level was significantly higher in the ACS patients than in NC group and SAP patients.The levels of UA,FIB and LDL were related to the Gensini score,the UA and FIB were related to the number of impaired arteries.Conclusions There were different levels of UA and FIB in different types of CHD and LDL was related to the Gensini score of coronary artery stenosis.

Abstract:Aim To discuss the changes and significance of plasma fibrinogen and D-dimer in patients with acute cerebral.Methods The levels of plasma fibrinogen and D-dimer in 96 cases of acute cerebral infarction and 30 healthy controls were detected with Clauss method and immunoturbidimetry method,respectively.Results The levels of plasma fibrinogen and D-dimer were significantly higher than those in the control group(P<0.05).Moreover,with the aggression of acute cerebral infarction,the levels of fibrinogen and D-dimer were increasing,gradually(P<0.05).The levels of fibrinogen were positively correlated with the levels of D-dimer(r=0.423,P<0.05).Conclusion The levels of plasma fibrinogen and D-dimer are useful for the diagnosis and evaluation of disease severity.