Abstract:AimTo investigate the proliferation effects of overexpression integrin-linked kinase on human umbilical vein endotheial cells (HUVEC).MethodsHUVEC were infected by integrin-linked kinase (ILK) Ad-virus and induced ILK overexpression.HUVEC proliferation levels were measured by thiazolyl blue (MTT), flow cytometry and 5-ethynyl-2’-deoxyuridine (EDU) assay.Western blot was used to assess protein kinase B (Akt), endothelial nitric oxide synthase (eNOS) expression and phosphorylation levels.ResultsMTT assay, flow cytometry cycle, EDU assay all showed that cells proliferation were significantly higher in ILK overexpressed group than in the control group (p<0.01 or p<0.05).The levels of Akt and eNOS phosphorylation levels were also significantly higher than that in control group (p<0.05), but Akt and eNOS expression levels had no difference between the two groups.ConclusionsILK overexpression can promote HUVECs proliferation.This effect may be mediated through actvation of Akt and eNOS.

Abstract:AimTo observe the influence of the early growth response factor-1 (Egr-1) decoy oligodeoxynucleotide (decoy ODNs) on the expression of Egr-1 and cysteine-rich protein 61 (CYR61/CCN1) in rat vascular smooth muscle cells (VSMCs) cultured in vitro and investigate the mechanism about Egr-1 decoy ODNs inhibiting VSMCs migration.MethodsCultured the VSMCs in vitro and transfected Egr-1 decoy ODNs.Detected the mRNA level and the protein expression of Egr-1 and CCN1 using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical method at the different time points of normal control group (normal cultured cells) and the decoy group (transfected Egr-1 decoy ODN).The migration lenth of each group was observed by wound-healing assay.ResultsThe results of RT-PCR and cell immunohistochemistry analysis showed that there was certain expression of Egr-1 and CCN1 mRNA and protein in these three groups.At the different time points, the level of Egr-1 and CCN1 mRNA and protein were decreased obviously in the decoy group compared with the normal control group (p<0.05).Wound-healing assay showed that the migration length of the control group, decoy and decoy SCR was 105.23±7.81 μm, 58.65±12.68 μm and 106.47±7.60 μm respectively.The migration length of the decoy group is much shorter than the other groups (p<0.01).ConclusionTransfecting the decoy ODNs of Egr-1 to the arterial VSMCs of rat in vitro can decrease the expression of Egr-1 and CCN1, to achieve the purpose of inhibiting cell migration.

Abstract:AimTo investigate the role of cAMP response element binding protein (CREB) in endothelial cells injury induced by high glucose. MethodsThe endothelial cell injury model was induced by high glucose.ECV304 cells were divided into six groups: control group, high glucose group, PCI transfection group, PCI transfection＋high glucose group, PCI-CREB transfection group and PCI-CREB transfection＋high glucose group.The cell survival rate of ECV304 cells was detected by MTT method.Flow cytometry was used to detect the apoptosis rate.The MDA concentration was observed by TBA method and SOD activity was tested by xanthine oxidase method.ResultsCompared with control group, cell survival rate, SOD activity, MDA concentration and apoptosis rate in PCI-CREB transfection group and PCI transfection group were not significantly different (p>0.05).Compared with control group, both cell survival rate and SOD activity in high glucose group showed a significant decrease, however, apoptosis rate and MDA concentration increased significantly (p<0.05).Cell survival rate, SOD activity, MDA concentration and apoptosis rate in PCI transfection＋high glucose group was not significantly different compared with high glucose group (p<0.05).Compared with high glucose group, both cell survival rate and SOD activity in PCI-CREB transfection＋high glucose group showed a significant increase, however, apoptosis rate and MDA concentration decreased significantly (p<0.05). ConclusionOverexpression of CREB may protect endothelial cells from high glucose injury.

Abstract:AimTo investigate the effect of hypoxia postconditioning on mitochondrial reactive oxygen species (ROS) and cardiomyocyte apoptosis induced by hypoxia/reoxygenation.MethodsThe model of cultured cardiomyocytes with hypoxia/reoxygenation (H/R) was established and the cardiomyocytes were divided into 3 groups, including control group, H/R group (hypoxia 3 h and reoxygenation 6 h), and hypoxia postconditioning (PC) group (3 intermittent cycles of 5 min H/R immediately after hypoxia 3 h, then reoxygenation for 6 h).Cardiac mitochondria and sarcolemma were isolated by differential centrifugation.ROS was detected with fluorescent probes and cardiomyocyte apoptosis was detected with flow cytometry.The expressions of Bcl-2 and Bax proteins in the cardiac mitochondria and sarcolemma were measured by Western blot.ResultsBoth mitochondrial ROS and the number of apoptotic cardiomyocytes reduced significantly in PC group compared with H/R group.Bcl-2 levels increased while Bax levels decreased in cardiac mitochondria and sarcolemma of PC group.ConclusionPC attenuated ROS and cardiomyocyte apoptosis induced by H/R, which potentially mediated by upregulating the expression of Bcl-2 and downregulating the Bax in mitochondria and sarcolemma.

Abstract:AimTo investigate the effects of DNA enzyme targeting early growth response factor-1 （Egr-1) mRNA (ED5) on the expression of PAI-1 after vascular balloon injury and the mechanism of inhibiting neointimal hyperplasia.MethodsTo establish the intimal injury model of left carotid artery in rats.Ninty-six rats were randomly divided into four groups, sham group, MgCl₂ group, FuGENE6 group and ED5 group.Six rats were killed at 3 d, 7 d, 14 d and 21 d after operation.Vascular sections were stained with heamotoxylin and eosin to identify pathological changes of vascular by light microscope.Immunohistochemistry stain was used to test the expression of Egr-1 inarterial wall.ELISA kit was used to test the expression of PAI-1 gene in plasma.ResultsThe levels of Egr-1 were reduced after treatment with ED5, and there were significant differences between the ED5 group and the MgCl₂ and FuGENE6 group (p<0.01).The expression of PAI-1 reduced at every time point in ED5 group (p<0.01).Compared with MgCl₂ group and FuGENE6 group, the intimal and media thickness and the stenosis rate of vessel in ED5 group at every time points after operation was significantly lowered (p<0.01).ConclusionThe level of PAI-1 increase after balloon injury.ED5 can through

Abstract:AimTo observe the change of the distribution and expression levels of connexin 40 (Cx40) in right atrial myocardium of people with no-atrial fibrillation (no-AF) and patients with different types of atrial fibrillation (AF), and to study the associations among distribution and expression changes of Cx40 with AF subtypes, right atrium (RA) size and initiation and development of AF.MethodsThe total 23 samples of right auricular appendage were collected.Five were designed to no-AF group, six patients with paroxysmal AF (PX), five patients with persistent AF (PS) and seven patients with permanent AF (PM) served as AF groups.RA size by echocardiographic analysis was indexed to body surface area.Expression and distribution of Cx40 were examined by immunoconfocal microscopy.The mRNA content of Cx40 was measured by relatively real-time quantitative polymerase chain reaction (real-time PCR).And protein expression level of Cx40 was studied by Western blotting.ResultsRA size of different AF subgroups was higher than that in no-AF group (p<0.05 or p<0.01).Immunoconfocal microscopy showed that Cx40 was mainly localized to the intercalated disc of cardiomyocytes in no-AF group, whereas exhibited heterogenous distribution in AF groups.The mRNA and protein expression levels of Cx40 were increased in PX group compared with no-AF group, however, Cx40 reduced in PS group and even lower in PM group (p<0.05 or p<0.01).The protein expression of Cx40 correlated negatively with

Abstract:AimTo observe the effect of ligustrazine (tetramethylpyrazine,TMP) pretreatment on the inflammatory reaction of myocardial ischemia reperfusion (IR) injury in rats and to investigate the possible mechanism.MethodsForty-eight Sprague-Dawley rats were randomly divided into sham group, IR group, TMP pretreated group, NG-nitro-L-arginine methyl ester (L-NAME) group and TMP+L-NAME group.All hearts except those in the sham group were subjected to 35 min ischemia followed by 120 min reperfusion.The myocardial structure was observed under microscope.The activity of myocardial superoxide dismutase (SOD) and the content of malondialdehyde (MDA), myeloperoxidase (MPO), interleukin-1β(IL-1β) and nitric oxide (NO) were measured.The expression of endothelial nitric oxide synthase (eNOS) was detected by reverse transcription polymerase chain reaction and Western blot.ResultsCompared with the IR group, the myocardial tissues of the TMP group showed an increase in the SOD activity, a decrease in the MDA content, the MPO activity and the level of IL-1β.TMP pretreatment also increased the NO level and alleviated the neutrophil infiltration.The expression of eNOS mRNA and protein in the myocardium of rats in the TMP group increased significantly compared with that in the IR group.However, these effects could be significantly reversed by L-NAME which abolished the increase of NO production and eNOS mRNA and protein expression brought by TMP.ConclusionsPretreatment of TMP attenuated the inflammatory reaction of myocardial IR in rats.The cardioprotective mechanism of TMP may relate to its effects of increasing the expression of eNOS and the level of NO.

Abstract:AimThe effect of ezetimibe (EZE) on acetylation of sterol regulatory element binding protein-1c (SREBP-1c) in lipid-loaded cells derived from vascular smooth muscle cell (VSMC) and its mechanisms have been explored.MethodsThe cholesterol∶methyl-β-cyclodextrin (Chol∶MβCD) complexes were used to build VSMC-derived lipid-loaded cells.After treated by 30 μmol/L EZE, the level of acetylated SREBP-1c, the level of fatty acid synthase (FAS) protein expression in lipid-loaded cells were detected by western blot, and the interaction between SIRT1 and SREBP-1c were analyzed by co-immunoprecipitation, and the role of SIRT1 in deacetylation of SREBP-1c was investigated using shRNA-mediated protein knockdown.ResultsThe level of acetylated SREBP-1c, the level of FAS protein expression were increased, but the interaction between SIRT1 and SREBP-1c was significantly reduced in lipid-loaded cells.However, EZE could weaken all these effects induced by Chol∶MβCD in lipid-loaded cells.A further study reveals that effects of EZE in lipid-loaded cells could be abolished by SIRT1 shRNA transfection.ConclusionEZE inhibits hyperacetylation of SREBP-1c in lipid-loaded cells required for SIRT1.

Abstract:AimTo investigate the effect of hydrogen sulfide (H₂S) on volume-regulated chloride channel (VRCC).MethodsH9c2 cardiomyocytes were cultured and treated with sodium hydrosulfide (NaHS, a H₂S donor).Expression of ClC-3 protein and VRCC chloride current (iCl-VRCC) were measured by Western blot assay and whole cell patch clamp, respectively.ResultsWhen H9c2 cardiomyocytes were placed in the isotonic solution, ICl was slightly activated.Hypotonicity obviously enhanced iCl (p<0.01), which was statistically attenuated by hypertonicity (p<0.05).ClC-3 protein was expressed in H9c2 cardiomyocytes.Similarly to the effect of isohytonicity on the activation of VRCC, treatment with 400 μmol/L NaHS for 0～30 min significantly increased iCl-VRCC (p<0.01), which was also statistically attenuated by hypertonicity (p<0.05).However, treatment with 400 μmol/L NaHS for 0～30 min did not alter the expression of ClC-3 protein in H9c2 cardiomyocytes (p>0.05).ConclusionsBoth VRCC and ClC-3 protein were expressed in H9c2 cardiomyocytes.H₂S activated VRCC in a ClC-3-independent manner.

Abstract:AimTo investigate the effects of hypoxia on the expression of Cx43 and apoptosis in hypertrophic rat cardiomyocytes.MethodsCardiomyocytes from newly born rats were cultured after digested in trypsin. The apoptosis rate of cardiomyocytes induced by hypoxia was evaluated by a modified Hoechst33258 fluorescent staining method.The expression of the Cx43 protein were detected by Western blot and fluorescent staining method.ResultsCompared with the control group, the apoptosis rate of hypertrophic cardiomyocytes induced by 24 h hypoxia was increased (15.42%±1.62%vs 2.56%±0.48%).The expression of Cx43 protein of hypertrophic cardiomyocytes was obviously decreased in comparison with the control group.Hypoxia could significantly down-regulate the Cx43 expression in hypertrophic cardiomyocytes than that of the normal cardiomyocytes (p<0.01). ConclusionsHypoxia can significantly increase the apoptosis rate and down-regulate the Cx43 expression in hypertrophic cardiomyocytes.These may be involved in the mechanisms of electrophysiology remodeling and malignant arrhythmia of hypertrophia myocardium induced by hypoxia.

Abstract:AimTo investigate the effects of puerarin on ventricular function and calcium levels in diabetic rats.MethodsForty SD rats were randomly divided into four groups: the control group, the diabetic model group, the puerarin low dose group and high dose group.The cardiac function, ventricular mass index, heart weight index and calcium levels were respectively measured.ResultsCompared with diabetic model group, in the puerarin low dose group and high dose group cardiac function was significantly improved, ventricular mass index, heart weight index and calcium levels were decreased, but the puerarin high doses group improved more significantly.ConclusionsPuerarin can improve cardiac function, ventricular mass index, heart weight index in diabetic rats, and its mechanism may be related to intracellular calcium levels.

Abstract:AimTo investigate the role of cardiac rehabilitation on improving exercise capacity and ameliorating emotional disorders in coronary artery bypass grafting patients.MethodsFifty-two patients who had to wait for coronary artery bypass grafting (CABG) in the 1st hospital of China medical university were randomly divided into cardiac rehabilitation group(n= 27) and usual care group(control, n=25).All of the patients received conventional therapy,and the rehabilitation group was treated by gradually increased exercise and psychological intervention.6 min walk test,left ventricular ejection fraction(LVEF), hamilton anxiety scale(HAMA), self-rating depress scale(HAMD) were measured in preoperative and on 15th postoperative day.ResultsOn 15th postoperative day,compared with control group, 6-min walk distance of rehabilitation group was significantly higher（298.15±59.24 m vs 220.80±54.25 m, p<0.01）, and the scores of HAMA, HAMD significantly dropped（2.07±2.45 scores vs 5.48±3.32 scores,2.26±2.31 scores vs 6.36±3.81 scores,p<0.01）, rehabilitation patient’s postoperative LVEF was improved compared with preoperative（p<0.01).With futher analysis, the rehabilitation group showed a negative correlation between HAMA(r=－0.53,p<0.01),HAMD(r=－0.60,p<0.01) level and 6-min walk distance respectively.ConclusionsPsychological intervention combined with exercise treatment to patients undergoing coronary artery bypass grafting may drop levels of anxiety and depression, improve exercise capacity.It deserves further clinical applications.

Abstract:AimTo investigate the relationship between blood pressure load, blood pressure variability and tumor necrosis factor-α (TNF-a) in prehypertensives.Methods100 normotensives and 105 prehypertensives were recruited, while 110 hypertensives were enrolled.24 h ambulatory blood pressure monitoring was carried out in the three groups, blood pressure load and variability was computed and TNF-a was measured respectively.ResultsTNF-a, blood pressure load during 24 h, daytime and nighttime in prehypertensives were higher than that in normotensives while lower than that in hypertensives (p<0.01).Blood pressure variability during 24 h, daytime and nighttime in prehypertensives were higher than that in normotensives (p<0.05).The systolic blood pressure variability during 24 h and daytime and the diastolic blood pressure variability during 24 h and night in prehypertensives were lower than that in hypertensives (p<0.05).Pearson correlation analysis and multiple linear stepwise regression showed that the level of TNF-α was positively correlated with the systolic blood pressure load during 24 h and daytime, the diastolic blood pressure load during 24 h and nighttime, the systolic blood pressure variability during 24 h and nighttime (p<0.05).The systolic blood pressure load during 24 h and nighttime as well as systolic blood pressure variability during 24 h and nighttime were risk factors for TNF-a (p<0.01).ConclusionsBlood pressure load and blood pressure variability was correlated with TNF-a.Inflammation was relevant to the level of blood pressure in prehypertensives.

Abstract:AimTo evaluate the effect of clinical pathways on door-to-balloon (D2B) time, the curative effect of myocardial reperfusion in patients with ST-segment elevation myocardial infarction (STEMI).Methods100 patients treated with STEMI emergency percutaneous coronary intervention (PCI) clinical pathways were chosen randomly as the experimental group while 100 un-treated patients were chosen as the control group.The primary observation indexes are: D2B time, the number of cases achieving thrombolysis in myocardial infarction (TIMI) 3 flow, lowering of ST segment 90 min after PCI, the time to peak serum creatine kinase-mb (CK-MB) levels and in-hospital mortality rate.ResultsThe D2B time of experimental group was significantly shorter than that of control group (Median, 65 min vs 95 min, p<0.001), and the number of patients receiving primary PCI within the guideline-recommended 90 min was greater in the experimental group than in the control group (98% vs 65%, p<0.001).More patients achieved TIMI 3 flow (94% vs 81%, p<0.05) and lowering of ST segment 90 min after PCI (88% vs 67%, p<0.05) than those in control group.Time to peak serum CK-MB levels was evidently improved (7.8±0.5 vs 10.1±0.4, p<0.05) and in-hospital mortality rate of the experimental group was significantly less than the control group (2% vs 7%, p<0.001).ConclusionsClinical pathways were associated with a significantly shorter D2B time, the increase of the proportion of patients undergoing primary PCI within 90 min of hospital arrival.It could also help to significantly improve the myocardial microcirculation of the infraction area, evidently promote the curative effect of myocardial reperfusion in acute myocardial infarction and reduce the in-hospital mortality rate.

Abstract:AimTo investigate the association between the connexin37 (Cx37) gene C1019T polymorphisms and the risk of Kawasaki disease (KD) and that complicated with coronary artery lesion (CAL).MethodsA total of 45 patients with KD and 60 healthy controls were genotyped by using polymerase chain reaction-restriction fragment length polymorphism and agarose gel electrophoresis for the C1019T polymorphism in Cx37 gene.ResultsFor C1019T polymorphism in Cx37 gene, there were no significant differences between KD patients and the controls in genotype frequencies of CC, TC and TT and allele frequencies of C and T (χ²=1.411 and 0.003, both p>0.05); there were no significant differences between KD patients with CAL and that without CAL in genotype and allele frequencies (χ²=0.633 and 0.438, both p>0.05).ConclusionNo association was found between C1019T polymorphism in Cx37 gene and the risk of KD or its complication of CAL in this study.

Abstract:AimTo investigate the association of plasma homocysteine (Hcy) with arterial stiffness, measured as brachial-ankle pulse wave velocity (BaPWV) in essential hypertensive patients.Methods146 essential hypertensive patients were enrolled in the study.All the patients accepted physical examination, laboratorial checks, and coronary risk factors were recorded.They were classified into four groups according to interquartile interval of homocysteine.And they were also classified into two groups according to BaPWV.The differences among these groups were analyzed.ResultsThe linear correlation analysis showed that Hcy was more positively correlated with age, smoking, systolic blood pressure (SBP), pulse pressure (PP), pulse pressure index (PPI) and BaPWV (r=0.278, 0.272, 0.608, 0.575, 0.460 and 0.351， p<0.05).BaPWV was more positively correlated with Hcy, SBP, PP and PPI (r=0.351, 0.322, 0.322 and 0.253, p<0.01).Multiple logistic regression analysis showed that PP and Hcy were both independent risk factors for BaPWV.ConclusionThis study showed a positive correlation between high levels of plasma Hcy and arterial stiffness measured as BaPWV in essential hypertensive patients.

Abstract:AimThe study was designed to investigate the association between plasma concentrations of the soluble interleukin-1 receptor family member ST2 (sST2) and the degree of coronary artery stenosis in patients with acute myocardical infarction (AMI).MethodsPlasma soluble ST2 levels were measured via enzyme-linked immunosorbent assay (ELISA) in 44 AMI patients within 12 hours of the onset.The levels of coronary stenosis were assessed by Gensini scores and numbers of diseased vessels were counted.The association between plasma soluble ST2 level and coronary artery stenosis degree was analysed by SPSS software.ResultsIn AMI patients, the plasma sST2 was increased significantly in high Gensini scores subgroup (p<0.001) and a positive relation was found between sST2 levels and Gensini scores (r＝0.772, p<0.001).The plasma sST2 level was also significantly higher in patients with multivessels compared with single vessel (p<0.01).ConclusionThe levels of plasma soluble ST2 was positively correlated with severity of coronary stenosis and may predict the degrees of coronary stenosis in AMI patients.

Abstract:AimTo study the relationship between serum myeloperoxidase (MPO), pregnancy-associated plasma protein-A (PAPP-A) and stability of carotid plaques in patients with cerebral infarction.MethodsThe study included 52 patients with cerebral infarction and with carotid plaques confirmed by high resolution MRI.Patients were classified according to the characteristic findings of plaques in MRI.Serum levels of MPO and PAPP-A were determined by ELISA.ResultsSerum levels of MPO and PAPP-A in patients of unstable plaques group and rupture plaques group were significantly enhanced compared to individuals of stable plaques group (p<0.05 or p<0.01).Serum levels of MPO in patients of unrupture plaques group were significantly enhanced compared to individuals of stable plaques group (p<0.05).Serum levels of PAPP-A in patients of rupture plaques group were significantly enhanced compared to individuals of unrupture plaques group (p<0.01).There was no significant difference in serum levels of PAPP-A between unrupture plaques group and stable plaques group.There was no significant difference in serum levels of MPO between unrupture plaques group and rupture plaques group.The level of serum MPO and PAPP-A was positively correlated with the stability of carotid plaques.ConclusionsThere was a relationship between the serum MPO, PAPP-A and the instability of carotid plaques.High serum levels of the markers may indicate that the plaques were vulnerable or ruptured.

Abstract:Lipoprotein(a) (Lp(a)) is an independent risk factor of cardiovascular disease.It is a big hurdle for us to study the Lp(a) pathogenicity due to its heterogeneity, which has been a hotspot in current medical research.This review was about the research progress of the relationship between Lp(a) heterogeneity and cardiovascular disease, which can provide more detail information regarding the pathogenic mechanism of Lp(a).

Abstract:Clinical studies showed inflammation plays a great role in all phases of the development of atherosclerotic lesions, from the fatty streak lesion formation to the eventual rupture of the vulnerable plaque.Inflammation biomarkers may reflect different aspects of the atherothrombotic process in relation to the stages of acute coronary syndrome, and may correlate with its severity.This review summarizes the literature on the role of biomarkers of the atherosclerotic plaque with a view to suggest some evidence of feasibility for identification of vulnerable plaques.These evidence may encourage patients with atherosclerosis with more perfect management, involved both primary and secondary prevention.

Abstract:Micro RNA (miRNA) are short about 19～22 nucleotides long noncoding RNA.They were isolated from plasmas from well-characterized patients with varying degrees of cardiac damage: acute myocardial infarction.More and more research show miRNA are novel biomarkers for cardiovascular diseases.Disease of the myocardial infarction is the most common cardiovascular defect and causes of adult morbidity and mortality.An early diagnosis may ensure immediate initiation of reperfusion therapy to potentially reduce the death rate.To compare the plasma of miRNA between healthy people and patients with acute myocardial infarction, it can be found that the plasma level of miRNA in patients with acute myocardial infarction are raised significantly.