Abstract:Aim The incidence of stroke in diabetes is increasing seriously,which is associated with endothelial dysfunction and increased inflammation in carotid artery.Our study is to investigate the effect of c-Jun amino-terminal kinase(JNK) specific inhibitor SP600125 on carotid endothelial function and monocyte chemotactic protein-1(MCP-1) expression in type 1 diabetes. Methods Animals were divided into five groups in this study.The 1st group was C57BL/6 wild type male mice;the 2nd group was INS2AKITA male mice,intraperitoneal injection with 0.9% normal saline(NS) each day for 8 weeks;the 3rd,4th,5th group were INS2AKITA male mice,intraperitoneal injection with SP600125 10 mg/kg,20 mg/kg,30 mg/kg respectively each day for consecutive 8 weeks.Then the mice were killed and sreum myeloperoxidase(MPO),malondialdehyde(MDA),nitric oxide(NO),total nitric oxide synthase(TNOS) were measured.HE staining was performed in carotid artery and immunohistochemistry was performed to investigate MCP-1 protein expression in endothelial cells of carotid artery.P-JNK,JNK,MCP-1 protein expression in carotid artery was measured by Western blot,signal transducer and activator of transcription-1(STAT-1) DNA binding activity was assayed by electrophoretic mobility shift assay(EMSA). Results Compared with wild type mice,serum MPO and MDA expression increased prominently(P<0.05),whereas TNOS,NO decreased and p-JNK/JNK,MCP-1 expression increased significantly in INS2AKITA mice(P<0.05);STAT-1 DNA binding activity increased prominently in type 1 diabetic group compared with that in control group(P<0.05).Compared with type 1 diabetic mice,after treatment with SP600125,MPO decreased and NO,TNOS increased and p-JNK/JNK,endothelial MCP-1 expression decreased(P<0.05).The effects of SP600125 on MPO,NO,TNOS,MCP-1 were increased accordingly as the dose of SP600125 increased.As the dose of SP600125 increased,MCP-1 protein expression decreased 21.82%,39.09%,68.18% respectively;STAT-1 DNA binding activity decreased 25.70%,33.20%,61.29% respectively compared with type 1 diabetic mice. Conclusions JNK specific inhibitor SP600125 could inhibit STAT-1 DNA binding activity,thus increase serum NO and TNOS,and improve carotid diastolic function.SP600125 could also decrease MCP1 and MPO in a dose-dependent manner.The results also showed JNK signal pathway is associated with carotid endothelial dysfunction and inflammatory expression via activation of STAT-1 in type 1 diabetes.Our study provides a novel pharmacologic agent to prevent the promotion of macrovascular complication in type 1 diabetes.

Abstract:Aim To investigate effects of angiotensin-(1-7) [Ang-(1-7)] on soluble Fas(sFas) and restenosis after percutaneous transluminal abdominal aorta angioplasty. Methods Twenty-four healthy New Zealand white rabbits were divided into 3 groups: sham group,control group and Ang-(1-7) group.Rabbits underwent percutaneous transluminal angioplasty in the abdominal aorta or sham surgery.Subsequently,an osmotic minipump was implanted for Ang-(1-7) or saline administered.Before and 3 days,7 days,14 days,28 days after angioplasty,the levels of sFas in plasma were measured by enzyme-linked immunosorbent assay(ELISA).Angiography and histomorphometric analysis were performed four weeks after angioplasty. Results Four weeks after angioplasty,the Ang-(1-7) group displayed a significant reduction in luminal diameter(4.11±0.10 mm vs 2.88±0.08 mm,P<0.05),neointimal thickness(207.51±16.70 μm vs 448.08±15.30 μm,P<0.05),neointimal area(0.266±0.009 mm2 vs 0.408±0.020 mm2,P<0.05),and restenosis rate(28.13%±2.74% vs 40.13%±2.74%,P=0.008) compared with the control group.Furthermore,the levels of sFas in plasma were elevated in the 3rd day after vascular injury both in control and Ang-(1-7) group,then the levels of sFas in Ang-(1-7) group gradually declined(1.023 3±0.063 4 μg/L),while the levels of sFas in control group achieved its peak in the 7th day(2.508 2±0.068 0 μg/L).The levels of angiotensinⅡ(AngⅡ) in plasma were raised both in control group and Ang-(1-7) group four weeks after angioplasty.However,the level of AngⅡ after angioplasty has no significant difference between these two groups(0.183±0.015 μg/L vs 0.178±0.015 μg/L,P>0.05). Conclusion Angiotensin-(1-7) attenuates neointimal formation and restenosis rate after angioplasty in rabbits and this effect might be conveyed through advancing the peak concentration of sFas in plasma,which could promote apoptosis of excessive proliferated vascular smooth muscle cell and achieving balance between proliferation and apoptosis as early as possible.

Abstract:Aim To study the effect of Amlodipine on CD40 expression in HUVEC-12 endothelial cells induced by oxidized low density lipoprotein(ox-LDL) to explore the new antiatherosclerotic role of Amlodipine. Methods The experiment groups included 6 groups: control group,10.0 μmol/L Amlodipine alone treated group,ox-LDL group,and different concentration(0.1,1.0,10.0 μmol/L) of Amlodipine pretreated for 1 h and treated with 50 mg/L ox-LDL for 24 h group.RT-PCR was used to detect the mRNA expression of CD40 and western-blot was used to detect the protein expression of CD40. Results RT-PCR and western blot showed that with ox-LDL and Amlodipine treatment,both mRNA and protein expression of CD40 were greatly decreased in a concentration-dependent manner. Conclusion Amlodipine can have a significant effect on down-regulating CD40 expression induced by ox-LDL.

Abstract:Aim To investigate the effect of Shexiang Baoxin Wan on cell proliferation,apoptosis and expression of inflammatory factors in human umbilicial vein endothelial cells(HUVEC) induced with hydrogen peroxide(H2O2). Methods Collagenase I was used to digest and isolate HUVEC and then cultured.Bromodeoxyuridine(BrdU) enzyme-linked immunosorbent assay(ELISA) was used to assay proliferation of HUVEC in vitro.Cell apoptosis was determined by terminal dUTP nick end labling(TUNEL).Expression of monocyte chemotactic protein-1(MCP-1),interleukin-6(IL-6) and nuclear factor-kappa B(NF-κB)-P65 mRNA was detected by reverse transcription polymerase chain reaction(RT-PCR).Western-blotting was performed to detect the expression of NF-κB-p65. Results(1)Compared with control,cell growth and proliferation was significantly lower in H2O2 group.The serum of Shexiang Baoxin Wan could inhibit the HUVEC proliferation declined by H2O2 in concentration-dependent manner.Apoptosis was higher in H2O2 groups than that in control group and reserved to normal in 1 g Shexiang Baoxin Wan.(2)Compared with control,the level of MCP-1,IL-6 and NF-κB mRNA in H2O2 group was greatly increased.Shexiang Baoxin Wan could inhibit the expression of these inflammatory factors induced by H2O2 in concentration-dependent manner.The serum of 1 g Shexiang Baoxin Wan remarkly decreased the level of MCP-1,IL-6 and NF-κB-P65 mRNA.(3) Compared to control,the level of NF-κB-P65 was largely increased in H2O2 group,Shexiang Baoxin Wan could inhibit the expression of NF-κB-P65 in concentration-dependent manner.The serum of 1 g Shexiang Baoxin Wan greatly decreased the level of NF-κB-P65. Conclusion The role of Shexiang Baoxin Wan in protecting endothelial cell may be related to inhibiting the inflammatory factors induced by H2O2.

Abstract:Aim To investigate the effects of deuterium-depleted water(DDW) and liquor on lipid metabolism,blood coagulation and fibrinolysis activity in experimental hyperlipidemic rats. Methods Normal control and hyperlipidemia model groups were designed.Experimental hyperlipidemia rats were administration of liquor [(0.01 L/(kg·d)] or deuterium-depleted water for 90 d.The levels of plasma triglyceride(TG),total cholesterol(TC),high-density lipoprotein(HDL),and low-density lipoprotein(LDL)were determined,and prothrombin time(PT),activated partial thromboplastin time(APTT),tissue plasminogen activator(t-PA)activity,and plasminogen activator inhibitor-l(PAI-1) activity were measured.The pathological examination of the aorta structure and liver tissue was performed. Results Compared with model group,HDL and t-PA significantly increased(P<0.05) and PAI-1 significantly decreased(P<0.05)in DDW group. LDL and PAI-1 significantly decreased(P<0.05) and PT and t-PA significantly increased(P<0.05) in low-dose liquor group.Further,TC,HDL,and PT and t-PA in high-dose liquor group significantly increased(P<0.05).The TC,HDL,LDL,and PT and t-PA significantly increased(P<0.05) and TG and PAI-1 significantly decreased(P<0.05) in high-dose liquor with DDW group.The TC,TG,and t-PA significantly increased(P<0.05) and APTT and PAI-1 significantly decreased(P<0.05) in low-dose liquor with DDW group. Conclusions Blood clotting system and fibrinolysis system in the experimental hyperlipidemic rats were improved by DDW and liquor,respectively.The combined effects of liquor and DDW on these 2 systems in hyperlipidemic rats were similar to those of liquor.

Abstract:Aim To investigate the impact of hyperuricemia(HUA) on blood pressure and serum adiponectin(APN) and to explore the relationship between endothelial function and the change of serum APN induced by hyperuricemia. Methods Thirty six male Sprague-Dawley(SD) rats were randomly divided into 3 goups.HUA was induced in rats over 6 weeks by a high-yeast extract(YE) diet united intraperitoneal injection of potassium oxonate(OA) suspension(n=12).Allopurinol was administered simultaneously with the OA and YE in allopurinol-treatment group.Normal diet-fed rats were used as control.Systolic blood pressure(SBP) was measured.At the end of 6th week plasma uric acid were detected by automatic biochemical analysis.APN,endothelin-1(ET-1),nitrogen monoxidum(NO) were measured by ELISA and the expression of endothelial nitricoxide synthase(eNOS) in aortic tunica intima was assessed by immunohistochemistry. Results Compared with contol,levels of UA,ET-1,SBP were significantly higher in model group,but levels of APN,NO and expression of eNOS in aortic tunica intima were significantly lower.There was an increase of serum APN,NO and higher expression of eNOS in aortic tunica intima in allopurinol-treatment group,but a decrease of UA,ET-1 and SBP.APN was negatively related to SUA,ET-1 and SBP. Conclusion Decrease of serum adiponectin and nitic oxide,and increase of endothelin-1 levels may be involved in the hyperuricemia-induced hypertension in rats.

Abstract:Aim To evaluate left ventricular function by the parameter of excursion of left ventricular wall motion in type 2 diabetes mellitus(T2DM) patients. Methods 36 T2DM patients and 25 normal subjects were investigated by real-time three-diametional echocardiography in order to quantitatively analyse the distance and time of the excursion,then the result of the two groups were compared. Results The regional ejection fractior(REF) and the whole ejection fraction(EF) of the two groups were not significantly different(P>0.05).Whole ejection fraction(EF) of the two groups were not significantly different(P>0.05).Compared with control group the standard difference(ESD) and minimum distance(Emin) of the excursion T2DM patients were decreased(P<0.05),and the time of excursion(E%) was increased in T2DM patients group(P<0.05). Conclusions The distance and time of excursion could be helpful for sensitively assessing the early left ventricular dysfunction and myocardiac damage and quantitatively provide accuracy information for clinical preventment and therapy of myocardiac damage in T2DM patients.

Abstract:Aim Postprandial hypertriglyceridemia is an independent risk factor for coronary heart disease and atherosclerosis.This study was designed to explore the effect of once 1000 u coenzyme A(CoA) injection or capsule on postprandial triglyceride level after a high-fat meal in Chinese patients with fasting hypertriglyceridemia. Methods Ten healthy controls and 20 patients with fasting hypertriglyceridemia took a baseline high-fat meal(800 calories,50 g fat) after diet direction.And blood samples were collected at 0,2,4 and 6 h after the baseline meal.On the second day,all patients accepted a high-fat meal again and randomly took 1000 u CoA orally at 30 min before the meal or by intravenous drip immediately after the meal within 1 h.Blood samples were also collected at 0,2,4 and 6 h after the second meal.Blood lipids levels were assayed and percentages of increment in postprandial triglyceride level at 2,4 and 6 h after the meal were calculated. Results The postprandial serum triglyceride levels increased significantly at 2,4,6 h in patients while only at 2 h in healthy controls(P<0.05).Postp-randial serum total cholesterol,high density lipoprotein cholesterol and low density lipoprotein cholesterol levels did not change significantly when compared with their fasting levels,respectively.Taking all patients accepting CoA capsule or injection as a total(n=20),a significant decrease was found in percentage of increment in postprandial triglyceride level at 6 h after CoA treatment(62.4%±13.1% vs 42.1%±9.2%,P<0.05).Subgroup analysis showed that both postprandial triglyceride level at 6 h(4.63±2.40 mmol/L vs 3.80±2.08 mmol/L,P<0.05) and percentage of increment in postprandial triglyceride level at 6 h(62.8%±17.7% vs 39.0%±14.9%,P<0.05) significantly decreased after CoA injection treatment.However,decreases of postprandial triglyceride level and percentage of increment in postprandial triglyceride level did not reach statistical significance in patients taking CoA capsule. Conclusion This small-sample study showed that once 1000 u CoA injection immediately after meal effectively lower postprandially increased serum triglyceride level at 6 h after a high-fat meal in patients with fasting hypertriglyceridemia while once 1000 u CoA capsule did not significantly decrease postprandial hypertriglyceridemia.

Abstract:Aim To analyse the vascular risk factors and the relation with atherosclerosis between different degree periventricular white lesions(PVL) and deep white matter lesions(DWML) in advanced age people. Methods The severity of PVL and DWML was graded and the cognitive score,vascular risk factors,lacunar infarction numbers,carotid atherosclerotic plaque score,the cerebral blood flow velocity and pulsatility index(PI)of intracranial vessels of the different grade participants were analyzed. Results Both severe PVL and DWML had significant difference in cognitive score(P<0.05),while severe PVL had significant difference in morbidity rate of hypertension and diabetes mellitus(P<0.05),and DWML had significant difference in lipidemia and lacunar infarction number(P<0.05).Severe PVL manifested both large and small vessel atherosclerosis(P<0.05),and severe DWML was only found with large vessel atherosclerosis(P<0.05). Conclusions Severe PVL and DWML both induced cognitive impairment.The small vessel atherosclerosis and hypoperfusion may play roles in the pathogenesis of PVL,and DWML may be involved in lipid factors and lacunar infarction.

Abstract:Aim To investigate the relationship between serum levels of inflammatory factors,adiponectin and atherosclerosis in the patients with type 2 diabetic mellitus(T2DM). Methods 125 cases of T2DM patients were divided into two groups according to the carotid intima-media thickness(IMT) as follows: 65 cases of non-atherosclerotic diabetic patients and 60 cases of atherosclerotic diabetic patients.30 non-diabetic subjects were recruited as normal control.The clinical characteristics were collected for each patient.Serum fasting plasma glucose(FPG),glycation hemoglobin A1c(GHbA1c),fasting insulin(FINS) 2 hours postprandic plasma glucose(2hPG),lipid profiles levels were measured.Serum levels of adiponectin,intercellular adhesion molecule-1(ICAM-1),tumor necrosis factor-α(TNF-α) and high sensitive C-reactive protein(hs-CRP) were also detected.Insulin resistance index(HOMA-IR) was calculated. Results The serum levels of total cholesterol,triglyceride(TG),low density lipoprotein cholesterol(LDLC),FPG,2hPG and GHbA1c were significantly higher in the T2DM group than that of normal control group(P<0.05).And the serum levels of TG and LDLC were higher in atherosclerotic T2DM patients than that of non-atherosclerotic T2DM patients(P<0.05).The serum levels of TNF-α and hs-CRP and HOMA-IR in atherosclerotic patients were significantly higher than that of in non-atherosclerotic and normal control group(P<0.05).However,serum adiponectin level in arteriosclerotic group was lower than that of other two groups(P<0.05).There were negative correlation between adiponecin and IMT,ICAM-1,TNF-α,hs-CRP and HOMA-IR(r=-0.574,-0.635,-0.681,-0.768 and-0.752,P<0.01).There were positive correlation between HOMA-IR and ICAM-1,TNF-α and hs-CRP(r=0.657,0.706 and 0.688,P<0.01).TG,LDLC and HOMA-IR were risk factors of T2DM complicated with arteriosclerosis according to multiple regression analysis. Conclusion Inflammatory factors and adiponectin are involved in the pathogenesis of diabetic vascular complications.The protective effects of adiponectin may be performed through its anti-inflammation to improve insulin resistance and delay the development of atherosclerosis.

Abstract:Aim Observing the effect of pravastatin and folic acid on the carotid atherosclerosis. Methods Dividing 164 cases of acute cerebral infarction patients with carotid atherosclerosis and dyslipidemia randomly into three groups,including pravastatin group,pravastatin and folic acid group,and folic acid group,which have been given medication for 6 months respectively.And then make comparison of the 3 groups before and after treatment about the difference of the serum high sensitivity C-reactive protein(hs-CRP),lipid levels and carotid intima-media thickness(IMT),carotid artery plaque area change. Results By comparison,levels of the serum hs-CRP in the 3 groups decrease significantly(P<0.05),which situation can also be found in the lipid levels and carotid IMT,carotid artery plaque area change(P<0.05).The alteration of lipid levels is statistically significant(P<0.05).In particular,improvement of all the indicators in the pravastatin and folic acid group after treatment,is more obvious than the other two groups(all P<0.05).Conclusions The folic acid combined treatment with pravastatin can reduce serum hs-CRP levels,delay and reverse carotid atherosclerosis better than any single use of pravastatin or folic acid,since the two may have a synergistic effect.Therefore,it is worthy of further exploration in a large-scale,multi-center study.

Abstract:Aim To study the association between obstructive sleep apnea syndrome(OSAS) and cardiovascular risk factors in the male elderly. Methods 582 male elderly subjects were recruited and underwent standard polysomnography(PSG).Body mass index(BMI),neck circumference,waist circumference,hip circumference,waist to hip ratio,blood pressure,fasting blood glucose(FBS),high sensitivity C-reactive protein(hs-CRP),total cholesterol(TC),high density lipoprotein cholesterol(HDLC),triglyceride(TG),low density lipoprotein cholesterol(LDLC),free fatty acids(FFA) were measured.With respect to apnea-hypopnea index(AHI),the subjects were divided into OSAS patients and controls,furthermore,according to the value of AHI,OSAS patients were classified into mild,moderate and severe OSAS ones. Results OSAS patients had higher BMI,neck circumference,waist circumference,hip circumference,systolic blood pressure,FBS,hs-CRP,TG than controls.HDLC,average oxyhemoglobin saturation and minimum oxyhemoglobin saturation during sleep time was lower in OSAS patients than controls.The prevalence of metabolic syndrome was higher in OSAS patients than in controls.Severe OSAS patients had higher BMI,neck circumference,waist circumference,hip circumference,average and minimum oxyhemoglobin saturation during sleep time,however lower HDLC than mild and moderate patients.In Spearman's correlate analysis,OSAS and AHI were positively correlated with BMI,neck circumference,waist circumference,hip circumference,systolic blood pressure,FBS,hs-CRP and inversely correlated with HDLC.In multiple stepwise regression analysis,neck circumference was independently correlated with OSAS.Conclusion OSAS in the male elderly is associated with cardiovascular risk factors,which maybe induce the increased morbidity and mortality of cardiovascular disease.

Abstract:Aim To understand the character of distribution for hyperlipemia and hyperlipemia-related diseases in different population. Methods 3019 people who came to our hospital for clinical exanimation were divided into groups according to their sex and age and were measured for their blood level of triglycerin(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDLC),and low-density lipoprotein cholesterol(LDLC).Then these data and clinical diagnosis were analyzed. Results The elevation of TC level was accompanied by the increase in age.In male group,the TG level was increased first followed by decrease with increase of age.However,TG level was enhanced with increase of age in female group.The level of LDLC in male group with age below 40 was significantly higher than in female counterpart(P<0.05),however,there was no statistic difference in LDLC level between male and female group with age over 40(P>0.05).Level of HDLC was decreased with the increase of age in female group,but we did not find any relation between HDLC level and age in male group.Detection rate of hyperlipidemia in male(59.0%) was higher than that in female(33.5%),female with high TG,high TC,high LDLC detection rate peaked in the≥61 age group,male with high TC and high LDLC peak detection rate in the same group,but peak prevalence of high TG in the 41～50 age group; The high level of fatty liver,hypertension,diabetes,hepatic steatosis were more frequently found in male group(29%,19%,8%,19%)compared with female group(16%,18%,6%,12%).In contrast,Atherosclerosis was more frequently found in female group(18%) compared with male group(12%).It was positively correlated between hyperlipemia and fatty liver,hypertension,diabetes,hepatic steatosis(P<0.05). Conclusion The hyperlipemia and hyperlipemia-related disease were differently distributed in different population divided by age and sex.Our finding is very important for the early prevention,diagnosis and treatment of hyperlipemia and hyperlipemia-related diseases and also supplies an interesting clue for research in the cause of the hyperlipemia and hyperlipemia-related diseases.

Abstract:Aim To establish an available and convenient method to isolate and culture the rabbit bone marrow-derived endothelial progenitor cells(EPC) and compare the characteristics of the two different EPC. Methods Obtained 2 mL bone marrow from each shinbone of about 4 weeks old New Zealand rabbit,mononuclear cells(MNC) were isolated by Ficoll density gradient centrifugation method and planted in the first culture flask,after incubated for 48 h,collecting the suspended cells into the second flask,supplemented with vascular endothelial growth factor(VEGF) in order to induce cells differentiation into EPC.To compare the growing characteristics of the former and later cells,immunocytochemistry was used to examine the expression of the surfaces markers and immunofluorescence for function detection. Results Earlier obtained mononuclear cells adhered after planted 30 minutes,about 3 days late,cells became long spindle-shaped,a little bigger,and formed blood island-like clone and vessels-like formation.To the 10 days,vortex cells can confluence the whole flask,but had less proliferation(early EPC).The second adherent cells,oval shape,and appeared colony-forming by the 5～7th day after adhered.At last,cells confluenced like slabstone,which can transfer for 10 generations consecutively(late EPC).The second adherent EPC have a deficiency of CD133+obviously during differentiation,but a higher expression of CD34+.Most first adherent EPC can uptake Dil-ac-LDL and Ulex Europaeus agglutinin-1(UEA-1),that is similar to the function identification of the second adherent EPC. Conclusion The improved Ficoll density gradient centrifugation integrated with differential attachment technique can efficiently isolate and culture rabbit bone marrow-derived EPC,and the late EPC have a stronger growing ability.

Abstract:Th17 cells were identified recently as CD4+T cell lineages.Transforming growth factor β(TGF-β) and interleukin-6(IL-6) are required to induce the development of Th17 cells from naive T cells.Retinoid-related orphan receptor gamma-t(RORγt) is the key transcription factor in the differentiation of Th17 cells and IL-17 is the main effector molecule.Th17 cells/IL-17 play an important role in inflammation.Atherosclerosis(As) is a chronic inflammatory disease.Inflammation plays an important role in the occurrence and development progress of As.Animal models and clinical trials have found that Th17 cells/IL-17 play a role in As.This paper reviewed the research progress in recent years on the related aspects of the relationship between Th17 cells/IL-17 and As.

Abstract:The transforming growth factor-β(TGF-β) is superfamily of growth factors,which has similar structure and plays a pivotal role in holding the structure of the vascular wall and the development of atherosclerosis.The TGF-β mediates Smad signaling to regulate cell function.In Smad family,Smad3 plays an important role in embryo later stage.Recently,the application of gene knockout and siRNA interference,Smad3's physiological role is further clarified.It has been found to participate in TGF-β-mediated cell proliferation,apoptosis,immunity supervision and cardiovascular development.The studies suggested that the TGF-β/Smad3 signaling is a critical pathway in the process of atherosclerosis,and that might be a potential therapeutic target.