Abstract:Aim To explore the role of c-Jun N-terminal kinase （JNK） pathway in high glucose-induced cardiomyocyte injury and whether hydrogen sulfide （H₂S） protects cardiomyocytes against high glucose-induced injury by modulating JNK pathway. Methods H9c2 cells were treated with high glucose to establish a model of cardiac cell injury. CCK-8 was used to detect cell viability, intracellular level of reactive oxygen species （ROS） was measured by DCFH-DA staining and photofluorography, mitochondrial membrane potential （MMP） was tested by Rh123 staining and photofluorography, the activation of JNK protein was measured by Western blot assay. Results Glucose at high concentration induced obvious injury of H9c2 cells, leading to a decrease in cell viability, an increase in ROS generation and MMP loss, as well as upregulation of phosphorylated JNK expression Pretreatment with N-acethl-L-cystein （NAC）, a ROS scavenger or SP600125 （an inhibitor of JNK） for 30 min inhibited high glucose-induced cytotoxicity Pretreatment with 400 μmol/L NaHS （a donor of H₂S） for 30 min prior to exposure of H9c2 cells to high glucose blocked high glucose-induced injuries, evidenced by an increase in cell viability, decreases in ROS generation and loss of MMP, along with inhibition of phosphorylated JNK expression induced by high glucose. Conclusion Both ROS and JNK pathway mediate H9c2 cardiac cell injury induced by high glucose, H₂S may protect cardiac cells against high glucose-induced injury by inhibiting oxidative stress and JNK pathway.

Abstract:Aim To investigate the effects of siRNA recombinant lentivirus targeting CREB binding protein （CBP） gene on neointimal hyperplasia in rat carotid arteries after balloon injury and its possible mechanism. Methods Forty-eight male SD rats were randomly divided into four groups: Sham group, PBS group, CBP-siRNA-Lentivirus group and NC-siRNA-Lentivirus group to establish the model of balloon-injury in left common carotid artery, and the rats were sacrificed 28 days after injury and in vivo gene transfer. The expression of CBP and acetylated nuclear factor kappaB p65 （NF-κB p65） were determined by real-time PCR and Western Blot. Proliferating cell nuclear antigen （PCNA） protein expression was detected by immunohistochemistry. Meanwhile, morphometric analysis was used to measure neointimal hyperplasia. Results 28 days after operations, lentivirus siRNA targeting CBP markedly decreased CBP mRNA and protein expression compared with PBS and NC-siRNA-Lentivirus groups（P<0.05）. CBP gene silencing significantly reduced the neointimal area （0.108±0.008 mm² vs 0.238±0.022 mm² and 0.252±0.016 mm², P<0.05） and intima / media ratio （0.706±0.062 vs 1.483±0.136 and 1.497±0.137, P<0.05）. Furthermore, compared with PBS and NC-siRNA-Lentivirus groups, PCNA and acetylation of NF-κB p65 expression were both obviously downregulated in CBP-siRNA-Lentivirus group （P<0.05）. Conclusions Lentivirus-mediated CBP gene silencing could efficiently suppress neointimal formation in balloon injured rat carotid artery, and the mechanism was involved with downregulation of NF-kB p65 acetylation.

Abstract:Aim In order to investigate the relationship between integrin-linked kinase （ILK） and proliferative capacity of cardiac myocytes, we observed the expression of ILK and cyclin D1 in cardiomyocytes of Sprague-Dawley （SD） rats at different development stages （embryonic, newborn, adult and old）. Methods Hearts of SD rats at different development stages （fetal day 16, postnatal day 1, 3, 7, postnatal month 2, postnatal year 1, 1.5） were used to study. Real time polymerase chain reaction （real time PCR） and Western blot analysis were used to assess ILK and cyclin D1 expression. Results The expression of ILK in myocardial tissue decreased progressively with aging in SD rats. Expression of ILK was the highest in the fetal life （16 days gestation）, and the level of ILK tended to be stable at 2 month after birth. Expression of cyclin D1 was positively correlated with ILK content. Conclusions The altered ILK and cyclin D1 parallel normal cardiac development, and expression of cyclin D1 is positively correlated with ILK content. ILK may be correlated with proliferative capacity of cardiac myocytes.

Abstract:Aim To explore the effect of hyperhomocysteinemia （HHcy） on the changes of myocardial enzyme spectrum and its correlation with P53 gene. Methods 36 male ApoE^―/― mice were randomly divided into 3 groups （n＝12 for each group）: AopE^-/- control group, hyperhomocysteinemia group and intervention group. They were respectively fed with nomal diet, normal diet enriched 1.7% methionine and nomal diet plus 1.7% methionine 0.006% folate and 0.000 4% vitamin B₁₂ for 14 weeks. 12 healthy 5-week-old male C57BL/6J mice were taken as normal control group. The levels of serum homocysteine （Hcy） and myocardial enzyme levels in the four groups were measured by automatic biochemical analyzer. The content of serum oxidized low density lipoprotein （ox-LDL） was detected by enzymelinked immunosorbent assay （ELISA） the protein and mRNA expressions of P53 in the heart were detected by immunohis tochemical assay and real time polymerase chain reaction （RT-PCR）. Results The content of Hcy was significantly higher in ApoE^―/― control group, HHcy group and folate treatment group than normal control group, a remarkable increase was observed in the HHcy mice （P<0.01）, the ox-LDL level showed a parallel increase with the Hcy, the difference was significant （P<0.05）. The contents of creatine kinase-MB （CK-MB）, creatine kinase （CK）, hydroxy butyrate dehydrogenase （HBDH） and lactate dehydrogenase （LDH） were increased significantly in HHcy group while folate and vitamin B₁₂ decreased serum CK-MB, CK and HBDH levels compared with HHcy group. The results of RT-PCR and immunohistochemical assay exposed an up-expressed P53 mRNA and protein in HHcy group. The changes of P53 mRNA and serum CK levels had positive correlation （r＝0.706 1, P<0.01）. Conclusions Hyperhomocysteinemia could result in the changes of myocardial enzyme spectrum, the homocysteine-induced oxidative stress might be its major pathogenic mechanism. The up-expressed P53 might also be involved in its pathogenesis. Replenishment of folate and vitamin B₁₂ could significantly antagonize the detrimental effects of exogenous Hcy.

Abstract:Aim To investigate the effect of bone marrow mesenchymal stem cells（BMSC）transplantation on injured vessels endothelium repair and vascular restenosis after carotid balloon injury in rabbits. Methods 48 rabbits of carotid artery stenosis model were established and randomly divided into BMSC transplantation group （n24） and control group （n24）. BMSC were cultured in vitro and transfected with recombinant adenovirus-mediated enhanced green fluorescent protein gene. Balloon injured carotid artery of rabbits, meanwhile, BMSC（10⁷/kg）were infused into rabbits injured artery by external carotid artery in BMSC transplantation group, and the same amount of PBS solution were infused into the control group. 1 week after transplantation, BMSC homing were detected by immunohistochemical techniques. 2 weeks after transplantation, the expression of platelet-endothelial cell adhesion molecule（CD31）, α-smooth muscle actin and proliferating cell nuclear antigen（PCNA）were detected by immunohistochemical staining. 4 weeks s after transplantation, the incidences of the vessels stenosis were detected by carotid artery arteriography, the neointimal area and the ratio of the intima/media area were examined by hematoxylin and eosin staining. Results BMSC with GFP homing were detected in injured vessels intima at 1 week after BMSC transplantation. CD31 continues to have expression in intima of BMSC transplantation group, while the control group did not express. The expression of PCNA in BMSC transplantation group were decreased significantly compared with control group（23.43%±2.80% vs 50.49%±3.60%，P<0.05）. The expression of SM α-actin in BMSC transplantation group were elevated significantly compared with control group（0.437± 0.049 vs 0.197±0.032， P<0.01）. The neointimal area（0.103±0.022 vs 0.214±0.024，P<0.01）and the ratio of the intima/media area（0.771±0.096 vs 1.646±0.223，P<0.01）were significantly decreased in BMSC transplantation group than control group at 4 week. The incidences of the vessels stenosis were decreased compared with control group（39.64%±2.30% vs 63.31%±2.82%，P<0.05）. Conclusion BMSC transplantation can promote repairing of endothelial and phenotypic transforming of vascular smooth muscle cells after carotid balloon injury in rabbits and inhibits neointima hyperplasia. BMSC transplantation can also reduce the restenosis of injured vessels.

Abstract:Aim To investigate the effect of integrin β1 on foam cells formation by RNA interference （RNAi）-mediated silencing. Methods Short hairpin RNAs against mouse integrin β1 was constructed. The foam cell formation induced by oxidized low density lipoprotein （ox-LDL） was analyzed in mouse RAW264.7 cells which were divided into three groups,including control group,siRNANC group （with transfection of siRNA negative） and siRNA group. Formation of foam cells and contents of total cholesterol were detected, atomic force microscope was used to observe the caveolaes on cell membrane,expression of CD36 protein and mRNA were detected. Results Decreased integrin β1 expression led to reduce foam cell formation and contents of total cholesterol in cells. Meanwhile, there were no caveolaes on cell membrane and expression of CD36 protein and mRNA were both decreased. Conclusions Silencing of integrin β1 reduces foam cells formation. Integrin β1 modulated the capacity of macrophages to take up ox-LDL possibly via CD36-dependent pathway.

Abstract:Aim To investigate angiotensin Ⅱ （Ang Ⅱ） and oxidized low density lipoprotein （ox-LDL） regulating the expression of Smurf2 and collagenⅠin cultured rat vascular smooth muscle cells （VSMC）. Methods The mRNA expression was tested by RT-PCR. The protein expression was examined by Western blot. The protein expression of collagenⅠwas examined by ELISA. Results Compared with control group, Ang Ⅱ decreased Smurf2 expression （P<0.05） and enhanced collagenⅠexpression （P<0.05）, ox-LDL enhanced Smurf2 expression （P<0.01） and decreased collagenⅠexpression （P<0.05）. Ox-LDL concentration-dependently induced Ang Ⅱ-decreased Smurf2 expression （P<0.05） and collagenⅠ concentration. Conclusion Ang Ⅱ decreased Smurf2 expression in cultured VSMC. Ang Ⅱ may inhibit Smurf2 expression to improve collagenⅠ production. ox-LDL enhanced Smurf2 expression. ox-LDL may improve the expression of Smurf2 to suppress the production of collagenⅠ. ox-LDL concentration-dependently induced Ang Ⅱ-decreased Smurf2 expression, while decreased collagenⅠ expression.

Abstract:Aim　To　investigate　the　regulating　effects　of　P38MAPK　signaling　pathway　on　the　differentiation　of　the　bone　marrow-derived　stem　cells（BMSC）　induced　by　BMP-2　toward　cardiomyocyte-like　cells　and　to　determine　the　possible　mechanisms. Methods　BMSC　were　separated　and　cultured　in　vitro.　The　third　passage　cells　were　divided　into　3　groups:　control　group,　BMP-2　inducer　group,BMP-2+P38MAPK　blocker　SB203580　group.　The　morphological　observation　was　performed　by　the　inverted　phase　contrast　microscope,　the　immunohistochemical　technique　was　used　for　detecting　the　expression　of　cardiac　specific　troponin（cTnT）　and　connexin　43（Cx43）,Western　blot　was　used　for　examining　of　the　changes　in　p-P38MAPK/P38MAPK　levels　in　BMSC　after　induced　with　the　BMP-2. Results　Expression　of　p-P38MAPK　of　BMSC　could　be　observed　at　15　min　after　induction　with　BMP-2,　increased　obviously　at　30　min,increased　increasingly　from　60　min.　Compared　with　inducer　group,the　expression　of　p-P38MAPK　of　blocker　group　decreased　obviously（P<0.05）.　The　expression　of　cardiac　specific　troponin（cTnT）　and　Cx43　were　negative　in　control　group,　and　positive　in　inducer　group　and　blocker　group.　Compared　with　inducer　group,　Cx43　positive　cells　increased　in　blocker　group（P<0.05）. Conclusion　BMP-2　could　induce　differentiation　of　BMSC　toward　cardiomyocyte-like　cells,and　the　P38MAPK　sinaling　pathway　plays　a　negative　regulating　role　during　induction.

Abstract:Aim To investigate the effects of ouabain or atrial natriuretic peptide （ANP） on angiotensinⅡ secreted by artery smooth muscle cells （ASMC） in WKY rats and spontaneously hypertensive rats （SHR）. Methods The thoracic ASMC isolated from SHR and WKY rats were cultured and interfered with different concentrations of ANP or ouabain （1×10^－9 mol/L, 1×10^－8 mol/L and 1×10^－7 mol/L） respectively. The levels of angiotensinⅡ in culture-medium were measured by radioimmunoassay before and after 3, 6, 12, and 24 hours of using ouabain or ANP （1×10^－7 mol/L）. Results AngiotensinⅡ level in WKY rats was lower than that in SHR. After using ouabain, angiotensinⅡ level of two kinds of rats was remarkably increased, but in WKY rats angiotensinⅡ level reached the peak after 12 hours, while in SHR reached the peak after 6 hours. With the increasing concentration of ouabain, angiotensinⅡ level in WKY rats was increased in a concentration-dependent manner, and in SHR increased in a concentration-independent manner. After using ANP, the level of angiotensinⅡ of two kinds of rats was significantly increased, but in WKY rats reached the peak after 3 hours, and in SHR increased in a time-dependent manner. With the increase of concentration of ANP, the level of angiotensinⅡ went down in WKY rats, whereas went up in SHR. Conclusions Ouabain or ANP can promote the secretion of angiotensinⅡ of ASMC in both WKY rats and SHR, and the secretion peak of angiotensinⅡ induced by ouabain is in advance in SHR. The secretion of AngiotensinⅡ induced by ANP is significantly abnormal in SHR.

Abstract:Aim To investigate the role of calcium in tumor necrosis factor α（TNF-α）-induced cardiomyocyte hypertrophy. Methods The protein content was assayed with Lowry’s method . The cardiomyocytes volumes were measured by computer photograph analysis system. The protein synthesis was assayed with ［³H］-leucine incorporation method. ［Ca²⁺］i transient was measured by Till image system by cell-loading Fura-2/AM. Results 2-APB, a selective IP₃R inhibitor, and/or ryanodine, a selective RyR inhibitor, significantly suppress the elevation of the amplitude and the rates of the spontaneous Ca²⁺ transients induced by TNF-α in cultured ventricular myocytes from the neonatal rat. Nifedipine, an L-type calcium channels antagonist, had little effect on it. The increase of protein content, ³H-leucine incorporation and cell size induced by TNF-α were significantly suppressed by 2-APB, and/or ryanodine. Nifedipine had no effect on it. Conclusion TNF-α induced cardiac hypertrophy through increasing of intracellular Ca²⁺ in cultured ventricular myocytes from the neonatal rat. It was induced by both IP₃R and RyR, not by L-type calcium channels.

Abstract:Aim To compare the short-term effects of rosuvastatin and atorvastatin on serum lipids and markers of inflammation and endothelial function in patients with stable atherosclerosis. Methods Thirty-six patients with stable atherosclerosis were randomly assigned to receive 10 mg/day of rosuvastatin or 20 mg/day atorvastatin for 4 weeks. The change in the levels and patterns of lipoproteins, high-sensitivity C-reactive protein （hs-CRP）, Rho-associated coiled-coil-containing protein kinase activity（ROCK）, and brachial artery flow-mediated dilation （FMD） of the brachial artery were assessed before and after statin therapy. Results Rosuvastatin and atorvastatin both significantly lowered levels of total cholesterol, low-density lipoprotein cholesterol, triglycerides，and hs-CRP from baseline values, and increased levels of hs-CRP and FMD. Both statins inhibited ROCK（P<0.05）, the extent of inhibition was greater with rosuvastatin （P<0.05）. ROCK and FMD was correlated to each other（P<0.05）. But ROCK was not correlated with the levels of low density lipoprotein cholesterol（LDLC）. FMD was not correlated with the levels of LDLC adn hs-CRP. Conclusions Short-term treatment with either rosuvastatin or atorvastatin inhibits ROCK and improves endothelium dysfunction in patients with atherosclerosis. But rosuvastatin inhibits ROCK even more than atorvastatin.

Abstract:Aim To investigate the protective effection of atorvastatin on cardiovascular in maintenance hemodialysis patients. Methods The new import maintenance hemodialysis （MHD） patients （100 cases） were randomly divided into atorvastatin treatment group （n＝50） and control group （n＝50）. Before and after the study, hemoglobin, serum lipids, liver and kidney function, and markers of myocardial damage in patients of the two groups were detected with conventional method, dynamic blood pressure changes were also monitored. Before and after the study, changes of coronary vascular stenosis and calcification in patients of the two groups were analyzed by 64 row spiral CT coronary angiography. At the same time, the incidence of cardiovascular events in patients of the two groups were comparatively analyzed. Results In atorvastatin treatment group, before and after the study, no significant differences was found in vascular number of small coronary arteries stenosis≥50%, vascular number of small coronary arteries stenosis＜50%, and vascular number of no vascular stenosis, respectively, but in control group, at the end of the study, vascular number of small coronary arteries stenosis≥50% and vascular number of small coronary arteries stenosis＜50% were obviously increased, and vascular number of no vascular stenosis was distinctly decreased compared with that before the study （P<0.05）. At the beginning of the study, there was no statistically significant difference of the average level of coronary artery calcification between the two groups （P>0.05）. In atorvastatin treatment group, no significant increase was found in the average level of coronary artery calcification at the end of the study compared with that at the beginning of the study （P>0.05）, but in control group, the average level of coronary artery calcification was distinctly increased after the study, compared with that before the study, and compared with that at the end of the study in atorvastatin treatment group （P<0.05）. At the end of the study, the average level of myocardial injury markers was significantly lower than that at the beginning of the study in atorvastatin treatment group and at the end of the study in control group, at the same time, in control group, the rate of cardiovascular events was significantly higher than that in atorvastatin treatment group （P<0.05）. Conclusions The cardiovascular protective effect of atorvastatin calcium on the new import MHD patients was probably carried out through inhibition of coronary vascular calcification and/ or lipid lowering.

Abstract:Aim To investigate the possible mechanism of cerebral infarction in type 2 diabetes mellitus （T2DM） through identifying the changes of metabolic products in serum by applying the method of ¹H-nuclear magnetic resonance （¹H-NMR）. Methods Nineteen patients with cerebral infarction complication in T2DM, 25 T2DM patients without cerebral infarction and 29 healthy controls were included in our study. The micromolecular substances of each participant in serum were detected by 1H-NMR. The metabolic profile of each participant was obtained. The partial least square discriminant analysis （PLS-DA） was used to examine the difference of metabolic profile among groups. Results Based on the analysis of score plots of groups, the T2DM patients with cerebral infarction, T2DM patients without cerebral infarction and the group of healthy controls were clearly distinguished by applying PLS-DA. The characteristic metabolic products of patients with cerebral infarction complication in T2DM included very low density lipoprotein （VLDL）, low density lipoprotein （LDL）, glucose, 3-hydroxy butanoic acid, lactate, pyruvic acid, glycoprotein, leucine, valine, prostaglandin D₂, E₂, homocysteine, trimethylamine oxide （TMAO） and betaine. Conclusions The results of our study suggested that the metabolic disorders of lipids, carbohydrate and amino acid may play essential role in the development of T2DM with cerebral infarction. The elevated level of prostaglandin D₂, E₂ and homocysteine might play a role in the occurrence of cerebral infarction in T2DM. The change of TMAO and betaine which were caused by enteric dysbacteriosis, also might be related to the occurrence of cerebral infarction in type 2 diabetes mellitus.

Abstract:Aim To evaluate the long-term outcomes of myocardial infarction after coronary artery interventional therapy （PCI）. Methods 862 patients（mean age 58 years,range 27～82 years） who accepted PCI were followed up. According to the time of PCI, they were divided into three groups: group of emergency PCI, group of remedial PCI and group of elective PCI. Death, major adverse cardiovascular or cerebrovascular events and angina pectoris were observed. The relative risk of adverse events in different patients were analyzed by multivariate Cox analysis. Results The patients were followed up for 4.26 to 5.25 years（average 4.93 years）. Incidence rate of death and heart failure in group of elective PCI was higher than the other two groups. Prognosis of PCI was relative with age, smoking, diabetic, coronary artery and ejection fraction. Conclusion Multiple factors participate in the prognosis of PCI, and for the acute myocardial infarction,long-term outcomes of emergency and remedial PCI were better than that of elective PCI.

Abstract:Aim To evaluate the diagnostic accuracy of coronary artery stenosis with 320-slice dynamic volume computed tomography （320-DVCT） in comparison of selective coronary angiography. Methods Eighty-five patients with suspected coronary artery disease underwent 320-DVCT coronary angiography and selective coronary angiography subsequently. All available coronary segments were included in the analysis, regardless of size or image quality. Lesion with ＞50% diameter stenosis were considered significant. Image quality was analyzed, and the sensitivity, specificity, positive and negative predictive value of mild, moderate, severe coronary stenosis, occlusive coronary artery and ≥50% stenosis were calculated. Results The images acquired from 320-DVCT were excellent and sufficient and can be used to evaluate coronary stenosis. The sensitivity, specificity, positive and negative predictive value of 320-DVCT for detecting severe coronary stenosis, occlusive coronary artery and ≥50% stenosis were 90.9%, 99.2%, 89.6%, 99.3% and 98.6%, respectively, by mild stenosis, 92.8%, 98.9%, 90.9%, 99.2% and 98.3%, respectively, by moderate stenosis, 87.4%, 98.8%, 89.2%, 98.6% and 97.6%, respectively, by severe stenosis, 50.0%, 99.9%, 66.7%, 99.8% and 99.7%, respectively, by occlusive coronary artery, 94.1%, 97.2%, 89.2%, 98.5% and 96.6%, respectively, by ≥50% coronary artery stenosis. Conclusions 320-DVCT is a new, noninvasive approach of assessing the stenosis of coronary artery and can provide accurate and reliable information.

Abstract:Vascular disease is one of the common complications of diabetes mellitus including macrovascular disease and microvascular disease. Macrovascular disease （commonly refers to atherosclerosis） which leads to cardiac-cerebral vascular disease is a major cause of death in patients with type 2 diabetes mellitus. Although diabetic macrovascular disease has become one of hot research in the diabetic complications field, the underlying mechanism is still far from being completely elucidated. This review describes current situation and prospect concerning hyperinsulinemia and atherosclerosis and discusses the possible pathogenesis.

Abstract:As the classic drugs for treatment of atherosclerosis, statins’ cholesterol-independent or pleiotropic effects are attracting more and more attention. Recent experimental and clinical studies have indicated the close relationship between Rho kinase and the pleiotropic effects of statins. Moreover, many studies abroad have suggested that the inhibition of Rho kinase could be a promising target for the treatment of cardiovascular disease. In this review, we will summarize the structure and activation of the Rho kinase, discuss the relationship between Rho kinase and atherosclerosis, and illustrate the Evidence-Based evidences and mechanisms of statins’ inhibition of Rho kinase, meanwhile, this article will also prospect the therapeutic potential of Rho kinase in the treatment of atherosclerosis and vascular disease in future.

Abstract:Endothelial system is a dynamic balance system to maintain the normal flow of blood, which is very important to maintain local homeostasis. It has been confirmed that endothelial cell integrity of the structure and function are important to the normal vessel wall permeability barrier, immune defense, anticoagulant，antithrombotic and anti-inflammatory response. Microvascular endothelial cell damage is closely related to myocardial no-reflow, which can significantly reduce the survival rate of patients with no-reflow, being clinical treatment challenges. Studies have shown ischemia-reperfusion results in structure and function of endothelial cell injury, which promote the development of no-reflow. The corresponding clinical intervention measures in a certain extent can repair the damage of endothelial cells and improve myocardial no-reflow to a certain extent. The latest endothelial cell injurious mechanism and clinical treatment application are reviewed here, in order to provide new theoretical basis of prevention of myocardial no-reflow for clinical treatment.