Abstract:AimThe present study aimed to investigate the bio-function and mechanism of cellular repressor of E1A-stimulated genes (CREG) protein on the migration of vascular smooth muscle cells (SMCs).MethodsHuman wild-type and glycosylation mutant CREG proteins (named wtCREG or mCREG) were transfected and purified in human 293F cells.Human SMCs with CREG knocked-down expression (named OB2) were used to evaluate the effects of two kinds of recombinant CREG protein.The migration of OB2 cells was evaluated by wound-healing assay.The expression and activity was detected by Western blot and gelatin zymography.The differentiational marker proteins of SMCs were i- dentified to express by Western blot analysis.Furthermore, using soluble mannose-6-phosphate/insulin-like growth factor-2 receptor (M6P/IGF2R) fragments and M6P/IGF2R neutralizing antibody, the blocking analysis was finished by detecting the migration of OB2 cells.ResultsBoth wtCREG and mCREG (400 nmol/L) inhibit the migration of OB2 cells.Meanwhile, the expression and activity of matrix metalloproteinase-2 (MMP2) and matrix metalloproteinase-9 (MMP9) were identified to reduce in OB2 cells when treated with recombinant CREG protein.Reversely, the expressions of myocardin, smooth muscle α-actin (SM α-actin), myosin heavy chain (MHC) and caldesmin were detected to enhance obviously.Further blocking experiments using soluble M6P/IGF2R fragments and M6P/IGF2R neutralizing antibody suggest that two kinds of recombinant protein are adequate for modulating SMC migration by binding to domains 11-13.ConclusionThese data suggest that soluble CREG protein can exert its biological function via binding to cell surface M6P/IGF2R, and thereby provide novel insights into CREG modulation of SMC phenotypic switching from contractile to migration.

Abstract:AimTo investigate the effect of mitofusin2 (Mfn2) with the removal of protein kinase A (PKA) phosphorylation site (Mfn2-PKA(△)) on promoting the apoptosis of vascular smooth muscle cell (VSMC) and related signaling pathway.MethodsVSMC were transfected with adenovirus vector encoding Mfn2-PKA(△) or Mfn2 (Adv-Mfn2-PKA(△), Adv-Mfn2).The abundance of Mfn2-PKA(△) protein and Mfn2 protein were determined by Western Blot analysis using Mfn2 phosphorylated antibody.Laser confocal microscopy(LCMS) was used to observe the location of proteins.The effect of Adv-Mfn2-PKA(△) on the apoptosis of VSMC was explored by cell death ELISA.Western Blot were used to detect the expression of p-Akt.ResultsBoth Mfn2-PKA(△) and Mfn2 were located at the outmembrane of mitochondria.ELISA assay showed Mfn2-PKA(△) had no effect on promoting the apoptosis of VSMC compared with Mfn2 (p<0.01).Western Blot indicated that overexpression of Mfn2-PKA(△) gene could not down-regulate the phosphorylation of Akt (p<0.01).ConclusionMfn2-PKA(△) could not induce the apoptosis of VSMC.PKA phosphorylation site plays an important role in regulating the function of Mfn2 gene.

Abstract:AimTo investigate the effect of oxidized low density lipoprotein (ox-LDL) on ten-eleven-translocation 2 (TET2) expression and autophagy in human umbilical vein endothelial cells (HUVEC).MethodsAfter treatment with ox-LDL for 24 h, RT-PCR was used to detect mRNA expression of TET2.Western blot was used to detect protein expression of TET2 and autophagic marker Beclin 1 and microtubule-associated protein light chain 3 (LC3).HUVEC were transfected with TET2 siRNA, and then Beclin 1 and LC3 were detected by using Western blot.ResultsAfter treatment with ox-LDL for 24 h, the expression of TET2 was decreased with a concentration-depended manner both in mRNA and protein levels.The protein expression of Beclin 1 and the ratio of LC3Ⅱ/LC3Ⅰ were increased with response to ox-LDL concentration.Transfection with TET2 siRNA, the expression of Beclin 1 and the ratio of LC3Ⅱ/LC3Ⅰ were upregulated.Conclusionox-LDL decreased the expression of TET2, which might contribute to the increase of HUVEC autophagy.

Abstract:AimTo study the effect of simvastatin on cardiac fibrosis of rats with diastolic heart failure (DHF).Methods30 SD rats were randomly divided into three groups: control group, DHF group and simvastatin group.DHF model was produced by abdominal aortic coarctation in the rats.The rats in simvastatin group were given simvastatin 2 mg/(kg·d) by intragastric administration for 4 weeks, the others were treated with equal isotonic Na chloride.After 4 weeks, cardiac function and hemorheology were assessed by echocardiography and catheterization.HE and Masson stainings were applied to observe altered myocardial histopathology and myocardial fibrosis, respectively.ResultsIn DHF group, IVS, LVPWT, LVMI, E/A ratio, SBP, DBP, LVSP, LVEDP, fiber tissue， ventricular perivascular collagen area (CA), inflammatory cell invasion and collagen volume fraction (CVF) were increased.Tau was prolonged and LV-dp/dtmax was decreased.While in simvastatin group E/A ratio, IVS, LVPWT, LVMI, SBP, DBP, LVSP, LVEDP, inflammatory cell invasion, fiber tissue, CA and CVF were significantly decreased, prolonged Tau was shortened, and LV-dp/dtmax was increased compared with DHF group.ConclusionSimvastatin improves cardiac diastolic function in rats with DHF by ameliorating myocardial fibrosis.

Abstract:AimTo investigate whether the downregulation of heat shock protein 90 (HSP90) was involved in cardiac cell injury induced by serum-glucose deprivation (SGD).MethodsH9c2 cardiomyocytes (H9c2 cells) were treated with SGD to establish an in vitro model of ischemic cardiac cell injury.H9c2 cells were pretreated with 17-AAG (a selective inhibitor of HSP90) for 60 min before exposure of cells to SGD.Cell viability was detected by CCK-8.The intracellular level of free calcium was measured by Fluo-3AM staining and photofluorography.The expressions of HSP90 and glucose-regulated protein 78 (GPR78) were tested by Western blot assay.ResultsTreatment of H9c2 cells with SGD for 24 h significantly induced downregulation of HSP90 expression, overload of intracellular calcium, and upregulation of GRP78 expression, which is a marker of endoplasmic reticulum stress.Pretreatment with 17-AAG, a selective HSP90 inhibitor, not only aggravated SGD-induced decrease in the viability of H9c2 cells, but also enhanced SGD-induced overload of intracellular calcium and upregulation of GRP78 expression in H9c2 cells.ConclusionInhibition of HSP90 may be one of the key mechanisms, by which H9c2 cells were damaged by SGD.

Abstract:AimTo investigate the effect of niacin on expression of Caveolin-1 in adipocytes and adipose tissue from experimental atherosclerosis rabbits.Methods16 New Zealand white rabbits were fed with high fat diet for 8 weeks, and then randomly divided into two groups: hyperlipidemia group (maintained on high fat diet for 6 weeks, n=8) and niacin treatment group (the same fat diet plus 200 mg/ (kg·d) niacin for 6 weeks, n=8).Rabbits in control group (n=8) were fed with normal diet for 14 weeks.At the end of the study, all rabbits were sacrificed to detect the blood lipid changes and Caveolin-1 expression in adipose tissue by real-time quantitative PCR.The direct effect of niacin on Caveolin-1 mRNA expression in primary rabbit adipocytes was assayed.ResultsCompared with control group, rabbits fed with high fat diet showed higher serum levels of total cholesterol, triglyceride and low density lipoprotein cholesterol, more aorta endometrial thickness and plaque area, less Caveolin-1 expression.Compared with hyperlipidemia group, rabbits treated with niacin showed less serum levels of total cholesterol, triglyceride and low density lipoprotein cholesterol, less aorta endometrial thickness and plaque area and more Caveolin-1 expression.In vitro study, niacin dose-dependenfly stimulated Caveolin-1 expression in cultured adipocytes.ConclusionsNiacin could improve the lipid pro- file and stabilize atheromatous plaque, which may be related to up-regulating the expression of Caveolin-1 in adipose tissue or adipocytes.

Abstract:AimThe aim of this study was to investigate whether ezetimibe has a protective effect on β-cell function in diabetic state.Methods8-week-old db/db mice were randomized to ezetimibe 10 mg/(kg·d), or placebo.The age-matched nondiabetic littermates db/m mice were treated with placebo as the controls.After 6 weeks’ treatment, intraperitoneal glucose tolerance test and immunohistochemical staining for insulin were performed.Islet perifusion were performed to evaluate kinetics of insulin release by using islet perfusion system.ResultsFasting blood glucose (FBG), total cholesterol (TC), low density lipoprotein cholesterol (LDLC) were lower in ezetimibe group than that in db/db group after 6 weeks’ treatment.Ezetimibe provided an improvement of glucose tolerance and greatly rescued islet β-cell mass.Ezetimibe treatment also improved first-phase insulin response.ConclusionEzetimibe treatment effectively improved glucose tolerance and β-cell function by significantly restored first-phase insulin secretion on db/db mice.

Abstract:AimTo explore an efficient method to establish a rat model of atherosclerosis using vitamin D₃ (VitD₃)and atherogenic diet.MethodsForty healthy Wistar rats were randomly divided into 4 groups, ie, a control group receiving standard diet, three model groups receiving atherogenic diet for 12 weeks.At the same time, model group 1 were injected VitD₃ 7.0×10⁵ IU/kg every two days before feeding for four times； model group 2 were injected VitD₃ 6.0×10⁵ IU/kg before feeding and 1.0×10⁵ IU/kg in the third, sixth and ninth week respectively after feeding; model group 3 were injected VitD₃ 6.0×10⁵ IU/kg before feeding and in the third,sixth and ninth week respectively after feeding.The body weights of all rats were recorded between 12 weeks.After 12 weeks, total cholesterol(TC), high-density lipoprotein cholesterol(HDLC), low-density lipoprotein cholesterol(LDLC), and triacylglycerol(TG) were measured.HE staining was used to verify the atherosclerotic lesion in aorta.ResultsCompared with the control group, the body weights of model groups were seriously decreased (p<0.01), especially model group 3 and its high death rate.The serum levels of TC and LDLC in model groups were obviously increased than the control group.The numbers in three model groups producing typical atherosclerotic lesion in aorta were 3, 8 and 5 respectively.ConclusionsIt was an effective method for establishing a rat model of atherosclerosis by feeding atherogenic diet and injecting VitD₃, and the suitable dose of VitD₃ was 6×10⁵ IU /kg before feeding and 1.0×10⁵ IU /kg in the third, sixth and ninth week respectively after feeding.

Abstract:AimTo investigate the effect of tail interacting protein of 47 kDa (TIP47) on lipid-droplets (LDs) formation and triglyceride accumulation in Macrophages.MethodsWestern blot was performed to detect the effects of TIP47 siRNA on the expression of TIP47 and adipose differentiation-related protein (ADRP).Oil-red O staining and lipid analysis were employed to measure the formation of LDs and accumulation of triglycerides in Macrophages.Results TIP47 siRNA clearly inhibited TIP47 protein expression, but did not affect the expression of ADRP protein.Furthermore, TIP47 siRNA suppressed ox-LDL induced LDs formation and triglycerides accumulation in macrophages.Conclusions The results indicates that TIP47 should be preventive or therapeutic target for diseases caused by excessive lipid accumulation through suppression of foam cells formation, such as atherosclerosis.

Abstract:AimTo explore the correlation between arterial elasticity indexes and serum high sensitivity C-reactive protein (hs-CRP) and matrix metalloproteinase-9 (MMP-9) in patients with coronary heart disease.MethodsLarge arterial elasticity index (C1) and small arterial elasticity index (C2) were measured with CVProfilor DO-2020.Serum hs-CRP and MMP-9 were measured with ABC-double-antibody sandwich ELISA.ResultsThere was significant difference between coronary heart disease group and control group in C1, C2, MMP-9 and hs-CRP.Logistic regression analysis showed that male, smoking, low density lipoprotein cholesterol, C2, history of hypertension, hs-CRP were independent risk factors for coronary heart disease.Significant inverse correlation was noted between C1 and hs-CRP (r＝－0.51), MMP-9 (r＝－0.49), and between C2 and hs-CRP (r＝－0.69), MMP-9 (r＝－0.55) (all p<0.01) in patients with coronary heart disease.Multiple linear regression analysis showed that independent risk factors for C1 included age, pulse pressure, systolic blood pressure, hs-CRP, low density lipoprotein cholesterol and MMP-9, and independent risk factors for C2 were age, pulse pressure, fasting blood sugar, hs-CRP and low density lipoprotein cholesterol in patients with coronary heart disease.ConclusionsThe present study showed that there was significantly negative correlation between arterial elasticity indexes and serum MMP-9 and hs-CRP in patients with coronary heart disease.The in- flammation may have important impact on arterial elasticity.

Abstract:AimTo discuss the correlation between leptin receptor (LEPR) gene polymorphism and patients of the Han nationality who suffer from hypertension and obesity in Yunnan region.MethodsSelect 283 hospitalized patients of the Han nationality who suffer from hypertension and obesity in yunnan region as hypertension group and select 153 healthy persons of the Han nationality as normal control group at the same time.According to body mass index (BMI) ≥28 kg/m², the hypertension group is divided into a group of 162 patients suffering from hypertension and obesity and a group of 121 patients suffering from hypertension but without obesity.Use polymerase even response-restriction fragment length polymorphism (PCR - RELP) method for determining Gln223Arg and Lys109Arg LEPR gene polymorphism, ELISA determination leptin levels in blood.Carotid intima-media thickness（CIMT） were measured by ultrasonography in all subjects.ResultsThe data of genotype frequency of Gln223Arg variant in leptin receptor gene between the case group and the control group had a significant difference (p<0.01).The data of genotype frequency of Lys109Arg variant in leptin receptor gene between the case group and the control group had no significant difference.Gln223Arg alleles LEPR gene frequency of A showed significant difference between the patients of hypertension with obesity and the patients of hypertension without obesity（p<0.01）.And patients of hypertension with obesity group has a higher CIMT（p<0.05）.LEPR gene Lys109Arg frequency showed no significant difference between the patients of hypertension with obesity and the patients of hypertension without obesity（p=0.226）.ConclusionLEPR gene polymorphism Gln223Arg was possibly associated with hypertension complicated with obesity in Han population in Yunnan area.And A allele is the risk factor to atherosclerosis in the patients of hypertension with obesity group.

Abstract:AimTo compare three different test methods monitoring the situation of inhibited platelet function in patients with acute cerebral infarction taking aspirin orally by establishing receiver operating characteristic (ROC) curve models, and to provide guidance for individual antiplatelet therapy.MethodsFrom February 2010 to October 2011, 127 patients with acute cerebral infarction not taking antiplatelet agents before were enrolled.The rate of CD62P expression was detected with flow cytometry (FCM), urinary 11-dehydro-thromboxane B2 levels was determined quantitatively with ELISA, the AA induced platelet inhibition rate was performed with thrombelastography (TEG) before and after 7-day treatment with aspirin orally.The sensitivity, specificity, and the area under the ROC curve (AUROC) of the three methods were calculated to evaluate their effects.ResultsCD62P expression rate, urine 11-dehydro-thromboxane B2 levels and platelet aggregation rate were significantly declined after treatment in aspirin group, and the differences were statistically significant (p<0.01).After treatment CD62P, urine 11-dehydro-thromboxane B2 levels and AA-induced TEG, had a certain correlation.The sensitivity, specificity and AUROC of the three methods were as follows: CD62P 86.7%, 79.8% and 0.805；11-DH-TXB2 78.5%, 64.7% and 0.767；TEG-AA 81.7%, 87.3%and 0.984.ConclusionsTEG has a high clinical value on monitoring the efficacy of aspirin in patients with acute cerebral infarction, which can provide more individualized guidance for antiplatelet treatment.

Abstract:AimTo investigate whether von Willebrand factor (vWF) can predict endothelialization after coronary stent implantation.MethodsOptical coherence tomography (OCT) images were obtained in 32 diseased coronary vessels with 34 stents in 30 patients with coronary artery disease who had no restenosis on follow up angiography at 12~18 months post drug-eluting stent (DES).Patients with complete endothelialization of the struts were considered as complete endothelialization group, patients with incomplete endothelialization of the DES struts were considered as incomplete endothelialization group, patients with angiographily diagonased nomal coronary were selected as the control group.For all the patients blood was drawed from elbow venous after operation, then plasma vWF levels of the three groups were tested.ResultsThe plasma vWF levels of incomplete endothelialization group were significantly higher than the complete endothelialization group and the control group (29.13±8.47 μg/L vs 13.81±1.04 μg/L and 12.83±1.13 μg/L,p<0.05), the plasma vWF levels had no significant difference in the complete endothelialization group and the control group (p>0.05).ConclusionsOur study confirms that the plasma vWF level has certain reference value in predicting endothelialization after coronary stent implantation.To some extent, the plasma vWF level can guide how long the antiplatelet therapy should be continued for the patients with coronary artery disease who had DES implanted.

Abstract:AimTo investigate the relationship between adiponectin and metabolic abnormalities in patients with acute cerebral infarction complicated by metabolic syndrome （MS）.Methods74 hospitalized patients with acute ischemic stroke were studied.All subjects were divided into MS group and non MS group with the criteria of MS published by international diabetes federation (IDF).For patient with MS, we divided it into three MS components group and four or five MS components group.The level of plasma adiponectin and serum TC, TG, LDL, HDL were detected by EILSA technology and the relationship between adiponectin and serum TC,TG, LDL,HDL were examined by Multiple linear regression analysis.ResultsThe patients with acute cerebral infarction complicated by MS showed lower levels of plasma adiponectin and higher levels of serum TG compared with control group (p<0.05).In four or five MS components group, the value of plasma adiponectin decreased obviously compared with control group and the levels of Serum TG, TC, LDL and HDL increased or decreased compared with control group (p<0.05).The result of Multiple linear regression analysis indicated that the levels of plasma adiponectin was significantly associated with TG inversely （b=－1.089，b’=－0.317，p=0.002）.ConclusionsThe value of plasma adiponectin in patients decreased obviously compared with control group.Plasma adiponectin was significantly associated with TG.

Abstract:AimTo study alterations in particle components of high density lipoprotein subspecies and activities of the key lipid transfer proteins for its metabolism in type 2 diabetic patients, the correlation between these alterations and related blood glucose indicators were also explored. Methods26 new-onset type 2 diabetic patients (T2DM group) and 25 healthy subjects (control group) were recruited. Plasma was both fasting and postprandial sampled for all subjects, then blood glucose, insulin and lipid spectrum were determined. High density lipoprotein 3 was separated by modified one-step precipitation method and the compositions were measured, then the major compositions of high density lipoprotein 2 were calculated. Activities of cholesterol ester transfer protein and phospholipid transfer protein was determined by enzyme fluorescence assay kits. Results (1)There were no differences in plasma high density lipoprotein cholesterol (HDLC) between T2DM group and control group (p＝0.579). (2)There were no significant differences in either cholesterol ester transfer protein or phospholipid transfer protein activity between T2DM group and control group (p>0.5). (3)The following particle components of high density lipoprotein 3 (HDL₃) in T2DM group were decreased significantly than those in control group (p<0.01): HDL₃C (0.60±0.17 mmol/L vs 0.75±0.22 mmol/L), phospholipids (PL, 1.14±0.19 mmol/L vs 1.29±0.22 mmol/L), free cholesterol (FC, 0.021±0.012 mmol/L vs 0.033±0.012 mmol/L), cholesterol ester (CE, 0.97±0.28 mmol/L vs 1.19±0.36 mmol/L).All the subjects’ FC and CE from H- DL₃ were significantly and negatively correlated with the following indicators (p<0.05): postprandial blood glucose (PBG), glycosylated hemoglobin (HbA1c), home-ostasis model assessment of insulin resistance (HOMA-IR). (4)The apolipoprotein AI (Apo AI) of HDL₂ in T2DM group was significantly decreased than those in control group (604.8±233.3 mg/L vs 755.4±198.0 mg/L, p<0.05), and the Apo AI of HDL₂ in all subjects was significantly and negatively correlated with PBG, HOMA-IR and plasma triglycerides (TG) (p<0.05). ConclusionsThe major components of HDL₃ and HDL₂ were decreased significantly in T2DM group than that in control group. We speculate that the size of HDL₃ and HDL₂ particle shrinks and might relate to hyperglycemia and insulin resistance.

Abstract:AimTo explore comprehensive intervention of coronary heart disease under Hospital-Community Health Integrated Management Mode, and to evaluate the effects.Methods583 patients with coronary heart disease from Shengze Jiangsu were randomly recruited into two groups: intervention group (287 patients) and control group (296 patients).Patients in the intervention group received comprehensive intervention for 1.5 years from July of 2010 to January of 2011, including health education and behavior, psychological and drug intervention; while patients in the control group received conventional treatment and education on relative knowledge of coronary heart disease.ResultsAfter 1.5-year intervention, the coronary risk factors awareness rate and healthy-behavior formation rate had shown significant change (p<0.01).The index of blood pressure and weight were significantly decreased (p<0.01).The level of blood glucose and lipid profiles were significantly improved (p<0.05).The cardiovascular events significantly decreased (p<0.05).ConclusionEstablishing comprehensive intervention of coronary heart disease under Hospital-Community Health Integrated Management Mode is entirely feasible.It serves as important exemplifications and is worthy of popularizing.

Abstract:AimTo investigate the prevalence of metabolic syndrome(MS) in Uighur and Han nationalities (≥50 years) in Xinjiang and compare the differences between the two groups of people.MethodsFrom July 2004 to August 2007,using hierarchical stochastic multistage cluster sampling, field survey and some combined investigation methods, a total of 8,382 people were investigated, including 3,603 Han people (male 1590, 2013 women), 4,779 Uighur people (male 2,347, 2,432 women), and the height, weight and abdominal circumference, blood pressure, blood sugar, blood lipid levels and other related indexes were tested.Results32.85% of Uighur and Han nationality had the prevalence of metabolic syndrome in the middle-aged (male 25.14%, female 39.07%, p<0.05); Han is 27.7% (male 22.53%, female 32.09%, p<0.05); Uighur is 41.03% (male 29.6%, female 50.30%,p<0.05); Uighur is higher than Han (p<0.05 ), and females than males, in which the prevalence of metabolic syndrome in Uighur men and women is higher than that of the Han nationality including both male and female (p<0.05).In two ethnic Uighur woman has the highest prevalence of metabolic syndrome.Prevalence of metabolic syndrome in the different age: from 50~59: Uighur 33.02%, Han 23.59%; from 60~69: Uighur 36.5%, Han 34.44%; above the age of 70: Uighur 27.65%, Han 28.3%.ConclusionsThere are inter-ethnic and gender differences between Han and Uighur in the prevalence of the metabolic syndrome in the middle-aged in Xinjiang.The prevalence of MS is higher in Uighur than in Han, especially in Uighur women.The prevalence of MS is the highest from 60 to 69.

Abstract:N-acetyl-seryl-aspartly-lysyl-proline （AcSDKP）is a kind of anti- fibrosis effects of short peptides, which is normally present in human plasma and circulating mononuclear cells. Recent studies found that AcSDKP was involved in some organ’s fibrosis, such as heart, lung, kidney and liver. AcSDKP could inhibit the target cells such as cardiac fibroblasts, lung fibroblasts, glomerular mesangial cells and hepatic stellate cells proliferation and collagen synthesis and expression. Through a variety of mechanisms including extracellular signal transduction AcSDKP can inhibit a target organ within the collagen deposition and reduce the causative factors leading to organ fibrosis.

Abstract:Dyslipidemia is one of the most important risk factors for atherosclerosis.From the 1960s, a large number of cholesterol-lowering clinical trials had laid a solid cornerstone of evidence-based medicine for the concept “the lower the better”.Statins can significantly reduce the occurrence of cardiovascular events by lowering low-density lipoprotein cholesterol (LDLC).However, there is still high residual cardiovascular risk in some patients who have achieved lower levels of LDLC, even when LDLC <70 mg/dL.High doses of intensive statin therapy can not completely solve the remnants of the residual risk.It is of great importance to solve the residual risk so as to further reduce the rates of cardiovascular events.This article aims to review the latest advances to clarify the relationship between high-density lipoprotein cholesterol (HDLC) and residual cardiovascular risk.

Abstract:Change of carotid intima-media thickness(CIMT) is earlier than the happen of plaque.As the early atherosclerosis (As) indicator it is increasingly attracting people’s attention, and has become a research hotspot at home and abroad.A large number of basic and clinical studies have shown that the traditional risk factors of As are similar to the risk factors of CIMT thickening.Recent international studies have found some other relevant factors can also cause increased CIMT.This paper make a brief overview about research progression of relevant factors of CIMT in recent years, in order to help the diagnosis and treatment of As.