Abstract:Aim To explore the effects of hydrogen sulfide on the ABCA1 expression and the formation of apolipoprotein. Methods HepG2 cells were treated with different doses of hydrogen sulfide. The cell viability was measured by MTT, the expression of protein was tested by Western blot, the expression of mRNA was detected by real-time PCR. Results Hydrogen sulfide had no influence on cell viability except for 400 μmol/L. At range of 4 to 24 h or 0 to 200 μmol/L, hydrogen sulfide could up-regulate the expression of ABCA1 and promote the serection of apolipoproteins in a dose-dependent manner but not in a time-dependent manner, compared with the control group. The mRNA of ABCA1 and apolipoproteins change in accord with the proteins. Conclusion Hydrogen sulfide up-regulates ABCA1 expression and promotes the formation of apolipoproteins.

Abstract:Aim To explore the effect of Calpain 1 on astragaloside Ⅳ （ASⅣ） inhibiting myocardial apoptosis induced by isoproterenol （ISO）. Methods 48 SD rats were randomly divided into 6 groups, 8 rats in each group: control group, ISO group, ISO＋propranolol 40 mg/（kg·d） group, ISO＋ASⅣ 20 mg/（kg·d） group, ISO＋ASⅣ 40 mg/（kg·d） group and ISO＋ASⅣ 80 mg/（kg·d） group. Administered groups received continually intragastric administration for 2 weeks, and ISO was intraperitoneal injected for 2 weeks in the day after that. 2 weeks later, TUNEL and electron microscopy experiments were performed to detect the myocardial apoptosis and mitochondrial changes respectively. Western blot was used to semi-quantify the expression of PARP1, mitochondrial Calpain 1 and apoptosis inducing factor （AIF）. Results Compared to control group, ISO group had a higher apoptosis percentage and mitochondrial swelling, cristae breakage were observed. In addition, the expression of poly ADP ribose polymerase 1 （PARP1）, mitochondrial Calpain 1 increased while mitochondrial AIF decreased. Compared to ISO group, ASⅣ group with different dose could decrease the apoptosis percentage and mitochondrial damage. Western blot results revealed that ASⅣ could decrease PARP1, mitochondrial Calpain 1 expression and increase mitochondrial AIF. Conclusions ASⅣ can protect myocardial apoptosis induced by ISO. The possible mechanism can be related to the inhibition of mitochondrial Calpain 1 expression, then reduce the release of mitochondria AIF into the cytosol and translocation to the nuclei.

Abstract:Aim To investigate the effects of exenatide on the development of aortic atherosclerotic lesions in diabetes ApoE^－/－ mice. Methods The male ApoE^－/－ mice were randomly divided into three groups: control group, diabetes group and exenatide group, all fed with high-fat diet. Diabetes group and exenatide group were injected with streptozocin （STZ） intraperitoneally to induce diabetes then infused with either placebo or exenatide for six weeks. At last their body weight, heart weight, blood glucose and serum lipids were measured. And the aortic atherosclerotic plaque area, plaque composition and plaque stability score were analyzed by using HE staining and immunohistochemistry. Meantime, the protein expression levels of poly（ADP-ribose）polymerase-1（PARP-1） and induced nitric oxide synthase （iNOS） were measured by Western Blot. Results Compared with control group, the diabetes group exhibited higher serum total cholesterol, triglycerides, blood glucose, aortic atherosclerotic plaque area and lower plaque stability score, and the expression of PARP-1 and iNOS increased. While compared with the diabetes group, the exenatide group exhibited lower serum total cholesterol, triglycerides, aortic atherosclerotic plaque area and higher plaque stability score, and the expression of PARP-1 and iNOS decreased. Conclusion Exenatide can suppress the area of aortic atherosclerotic lesions and stabilize the aortic atherosclerotic plaques through PARP-1 pathway in diabetes ApoE^－/－ mice.

Abstract:Aim To investigate the effect of alisol A 24-acetate on the phenotypic modulation of vascular smooth muscle cells（VSMC）as well as the expression of matrix metalloproteinase-9（MMP-9） in ox-LDL-induced rats vascular smooth muscle cells （VSMC）, and their correlation with ERK1/2 pathway. Methods VSMC isolated from the thoracic aorta of rats were induced by ox-LDL（50 mg/L） and intervened by alisol A 24-acetate （10 mg/L）. Immunocytochemistry was performed to detect the expression of SM22α protein.RT-PCR was performed to detect MMP-9 mRNA expression and Western blot was applied to detect the expressions of MMP-9, p-ERK and t-ERK. Results The contractive phenotypic specificity protein SM22α in the VSMC induced by ox-LDL was inhibited, VSMCs changed the phenotype from constriction to synthesis, the expression of phosphorylated ERK （p-ERK） and MMP-9 in ox-LDL group were elevated compared with the control group（P<0.05）. Alisol A 24-acetate.partially inversed the effects of ox-LDL on VSMC （P<0.05）. Conclusion Alisol A 24-acetate inhibited the phenotype transformation of VSMC induced by ox-LDL，and the mechanism maybe has correlation with ERK1/2 pathway.

Abstract:Aim To investigate the effects of calcium blocker nifedipine and diuretic hydrochlorothiazide on mechanical stretch stress mediated phosphorylation of ERK1/2 （p-ERK1/2） and expression of Ki67 in vascular smooth muscle cells （VSMC）. Methods Cultured quiescent VSMC were pretreated with nifedipine and hydrochlorothiazide respectively and subjected to treatment with mechanical stretch stress. Level of p-ERK1/2 in the treated cells was detected by Western blot and meanwhile Ki67 expression was detected by immunofluorescent staining. Results Compared with the negative control group, nifedipine or hydrochlorothiazide had no effects on p-ERK1/2 and Ki67 expression in quiescent VSMC, while mechanical stretch stress stimulation significantly increased levels of p-ERK1/2 and Ki67 expression, which was inhibited by nifedipine in a concentration-dependent manner, and synergistically enhanced by hydrochlorothiazide. Conclusions Hydrochlorothiazide synergistically promotes increased p-ERK1/2 and Ki67 expression in VSMC induced by mechanical stretch stress, which can be inhibited by nifedipine in a concentration-dependent manner. These results provide novel mechanisms for traditional antihypertensive drugs.

Abstract:Aim To construct and identificate recombinant adenovirus with siNrdp1 gene using AdMax system, and investigate the effect of Nrdp1 gene on cardiomyocyte hypertrophy. Methods Designing and synthesizing siRNA sequences targeting of Nrdp1 DNA, then cloned into the shuttle vector GV119 and homologous recombinated with adenovirus backbone plasmid AdMax in BJ5183 bacteria transfected HEK293 cells, and got adenovirus containing Nrdp1-siRNA gene through packaging. Real-time quantitative PCR and Western blot were used to detect Nrdp1 expression in primary rat neonatal cardiomyocytes. After adenoviral containing siNrdp1 transfection and angiotensin Ⅱ （AngⅡ） stimulation, real-time quantitative PCR was used to detect the expression of myocardial hypertrophy marker gene （ANF, β-MHC and Skeletal-α-actin） of rat neonatal cardiomyocytes. Results Digested PCR analysis and sequencing showed that interference Nrdp1 adenovirus was successfully constructed, and the titer of virus was 1.5E＋9 PFU/mL. Real-time PCR and Western blot indicated that the expressions of Nrdp1 mRNA and protein were greatly inhibited after infection in rat primary cardiomyocytes with recombinant adenovirus particles （P<0.001）. Nrdp1 gene silencing cloud significantly increase expression of AngⅡ induced cardiomyocyte hypertrophy marker genes including ANF, β-MHC and Skeletal-α-actin （P<0.01）. Conclusion The recombinant adenovirus vector containing the Nrdp1-siRNA gene was successfully constructed, which can effectively silence Nrdp1 gene and enhance AngⅡ induced cardiomyocytes hypertrophy in vitro.

Abstract:Aim To investigate the expression of Orai1 in the formation of atherosclerotic plaque of apolipoprotein E knockout （ApoE^－/－） mice. Methods 7 to 8 weeks old male ApoE^－/－ mice and wild-type C57BL/6J mice were selected and fed with fat rich diet. The animals were sacrificed consecutively after the mice were 20, 27 and 33 weeks old. Their aortas were taken for preparing the serial sections. The percentage of plaque area in luminal area was calculated after HE staining The percentage of collagen in plaque area was calculated after Masson staining Lipid content was analyzed after oil red O staining The percentage of smooth muscle cells expressing Orai1 was calculated after immunohistochemical staining. The dynamic expression of Orai1 during vulnerable plaque formation was quantitatively analyzed by using Western Blot analysis. Results Compared with the control group, the expression of Orai1 were markedly higher in ApoE^－/－ mice. As the age of ApoE^－/－ mice increasing, the expression of Orai1 increased significantly （P<0.05）. Conclusions Orai1 participates in the pathological process of atherosclerosis. The expression of Orai1 is up-regulated in the formation of atherosclerotic plaques.

Abstract:Aim To investigate the influence of transforming growth factor-α （TGF-α） on the differentiation capacity of human endothelial progenitor cells（EPC） and to explore its mechanism of action. Methods EPC were isolated from peripheral blood of healthy people, and cultured with complete medium containing 0, 1, 10 μg/L concentration of TGF-α. Morphological changes were observed in each group in EPC differentiation. By using the flow cytometry, the CD34⁺/CD133⁺ and CD31⁺/vWF⁺ positive cells were detected during the differentiation of cultured EPC with TGF-α. The expression of eNOS and Flk-1/KDR genes, the specific genes of EPC differentiated endothelial cells, were determinated by Real-time PCR. The NO content changes in each group were detected. The TGF-α receptor （EGFR） and vascular endothelial growth factor （VEGF） expression in EPC differentiation were tested by Western Blot. Results The cell morphology of EPC induced and cultured by TGF-α, could faster differentiate stem cells from the spherical to spindle cells.

Abstract:Aim To explore the effects and underlying mechanism of the exogenous hydrogen sulphide（H₂S） donor hydrosulfide（NaHS） on calcification （CAL） in human umbilical vein smooth muscle cell （HUSMC）. Method

Abstract:Aim To establish the swine model of myocardial infarction by percutaneous coronary artery embolism with gelatin sponge, and to observe the dynamic changes of early serum ischemia modified albumin （IMA） in swine with acute myocardial infarction （AMI）. Methods 16 Chinese mini-swines were via femoral artery puncture after anesthesia, and obtuse marginal branch was blocked by injecting gelatin sponge via micro catheter after coronary artery angiography. The dynamic changes of IMA were observed by continuously measuring serum IMA in the preoperative and postoperative 10, 30 min and 1, 2, 3, 4, 6, 8, 10, 12, 24, 48 h. The swines were sacrificed at 1, 3, 5, 7, 10 days after the AMI, and specimens of the myocardium were taken for histopathological examination. Results The 15 mini-swine models of myocardial infarction were established successfully, and AMI was confirmed by electrocardiogram, cardiac specimen and histopathological examination. IMA began to rise after 10 min and arrived the highest serum concentration in 6 h, and decreased to normal range in 48 h after AMI. Conclusions The method of swine AMI model established by percutaneous coronary artery embolism with gelatin sponge is scientifically reliable, simple, and repeatable. IMA is sensitive marker in the early of AMI, and dynamic detection of serum IMA level has certain clinical significance in early diagnosis of AMI.

Abstract:Aim To investigate if erythrocyte membrane cholesterol （EMC） precipitated the onset of the percutaneous coronary intervention （PCI）-related coronary slow or no reflow phenomenon, and to determine if EMC was a new predictor for this PCI-related coronary slow or no reflow in patients with non-ST elevated acute coronary syndrome （ACS）.

Abstract:Aim To compare the functional difference of endothelial progenitor cells （EPC） of high concentration Lp（a） （≥300 mg/L） coronary artery disease （HLPCAD） patients and low concentration Lp（a） （＜300 mg/L） coronary artery disease （LLPCAD） patients. Methods Differential adherence method was used to isolate EPC, Dil-ac-LDL swallowed and lectin binding was used for EPC identification. MTT was used to assay EPC survival and proliferation, modified Boyden chamber for migration, gelatin slide method for adhesion, a single cell hybridoma clones dish was observed and counted, and tubular structures formed on matrigel matrix length was measured. Results The numbers of circulating EPC in HLPCAD patients were significantly lower than those in LLPCAD patients （109.4±13.8 Cells/field vs.384.0±37.0 Cells/field, P0.0023）. MTT analysis showed that the OD value of LLPCAD group was 0.77±0.05, and the HLPCAD was 0.23±0.04 （P0.0018）, the apoptosis rate of HLPCAD EPC was significantly higher than that in LLPCAD group （14.9%±3.3% vs.4.1%±0.8%, P0.035）. The numbers of adhesion （25.3±4.6 Cells/field vs. 78.6 ± 6.8 Cells/field, P0.0030）, migration （22.0±2.6 Cells/field vs. 56.0 ± 4.9 Cells/field, P0.0037）, and clone-form units （2.4±0.4 number/field vs.11.0±1.3 number/field, P0.0003）, tubular structure formation （7.4±1.2 mm/field vs.33.3±2.6 mm/field, P0.0001） of HLPCAD patients were significantly reduced. Conclusions Function of HLPCAD EPC is seriously impaired while compared with LLPCAD EPC.

Abstract:Aim To investigate the changes of CD137 level in patients with acute coronary syndrome （ACS）.

Abstract:Aim To evaluate the effect of rosuvastatin medication with different doses on serum level of soluble lectin-like oxidized low-density lipoprotein receptor-1（sLOX-1）in patients with acute coronary syndrome（ACS）. Methods Patients（n80）with ACS were randomly assigned into rosuvastatin 10 mg group（40 mg loading dose and 10 mg/d,RSV10 group,n40）and rosuvastatin 5 mg group（5 mg/d, RSV5 group,n40）. All patients were accepted based on the clinical situation, including coronary revascularization, and aspirin, clopidogrel hydrogen, tirofiban, low molecular weight heparin, ACEI/ARB, β-blockers, calcium channel blockers, nitric acid esters drugs, diuretics and other basic drugs. Major adverse cardiac events（MACE）were observed during hospitalization, monitoring adverse drug reactions. Serum sLOX-1 levels of immediate admission on the third day were determinated using enzyme-linked immunosorbent assay and were compared between groups. Results Baseline characteristics were nearly identical in both groups. The density of sLOX-1 on the third day during hospitalization is 290.03±141.43 ng/L in RSV10 group, which remarkablely decreases in comparison with the value of 397.86±170.61 ng/L on admission. Reduce of the rate is 27.1%（t4.625，P<0.001）；The density of sLOX-1 in the third day during hospitalization is 300.03±135.31 ng/L in RSV5 group. The value of sLOX-1 declined significantly as well compared with 385.66±154.39 ng/L on admission，and the reduction is 22.2%（t3.329，P<0.05）； The comparison of the sLOX-1 density between the two groups on admission has no significance（t－0.335，P0.739） The reduction has no significance as well between the two groups（t－0.639，P0.525）. There is no difference in leukocyke count and neutrophil percentage，which actually decreased after treatment in both groups and RSV10 group decreased more significantly（t2.254，P<0.05）； The rate of MACE during hospitalization in RSV10 group is lower than RSV5 group（χ²4.588，P<0.05） There is no adverse drug recation during hospitalization in both two groups. Conclusion In clinical practice,early rosuvastatin treatment could significantly decrease the serum density of sLOX-1 in patients with ACS,obviously. Besides, the effect showed no dose dependent. Loading-dose and mild moderate-dose statin,which is relatively safe, could reduce the incidence of MACE during hospitalization.

Abstract:Aim To discuss the foreboding of avL lead T wave shape changes in ECG for the blocking of the coronary left anterior descending branch middle part in acute coronary syndrome. Methods Check the coronary angiography records and the electrocardiogram of the patients treated with coronary angiography in Shengjing Hospital in Huaxiang District of China Medical University from January 2009 to July 2010 due to acute coronary syndrome by information centre, in the 194 cases the average age was 65 years old, 140 cases were male. According to avL leads T wave shape, the patients were divided into inversion group and non-inversion group to analyze the relationship of avL lead T-wave and coronary artery disease by Logistic regression analysis. Results Compared with non-avL lead T-wave inversion group, the stenosis of middle left anterior descending artery was higher in T-wave inversion stenosis group （P<0.05）. T wave inversion and middle left anterior descending artery stenosis degree were independently associated（P<0.05）. Conclusion

Abstract:Aim To investigate the relationships between the changes in serum visfatin and coronary artery lesions, cardiac function in patients with acute myocardial infarction, and their chinical significance. Methods Acute myocardial infarction （AMI） patients with emergency percutaneous coronary intervention were picked out as test group （AMI group, n＝30）, including twenty-two men and eight women whose average age was 54.77±10.09. The degree of coronary artery lesions was assessed by the number of coronary artery lesions and Gensini integral. Meanwhile, the patients undergoing coronary angiography without any artery lesions were as control group （n＝30）, including twenty men and ten women whose average age is 54.07±11.07. Serum level of visfatin was measured by enzyme-linked immunosorbent assay （ELISA） at pre-operation and 24 hours after operation Left ventricular ejection fraction （LVEF） and left ventricular end-diastolic diameter （LVEDD） were measured by echocardiography. Results Serum levels of visfatin at both time points （pre-operation: 80.82±7.63 μg/L, 24 hours after operation: 91.96±7.37 μg/L） were significantly higher than that of control group （19.32±4.37 μg/L） （P<0.01）. Compared with pre-operative level, serum visfatin level was significantly increased at 24 hours after operation （P<0.01）. Serum levels of visfatin from different numbers of coronary artery lesion were: single-lesion: 72.85±2.56 μg/L, double-lesions: 82.24±5.77 μg/L, multi-lesions: 88.22±6.07 μg/L） Serum levels of visfatin from different stenosis degree were: mild: 74.58±4.40 μg/L, moderate: 80.13±4.71 μg/L, severe: 87.57±6.39 μg/L The differences between groups were statistically significant （all P<0.05）. Compared with control group, LVEF in AMI group was significantly lower （P<0.01），but LVEDD wasn’t significantly different. Serum level of visfatin was positively correlated with coronary artery lesion counts, coronary artery lesion Gensini integral （r＝0.754, r＝0.672, P<0.01）, whereas was negatively correlated with LVEF （r＝－0.459, P<0.01） in AMI group. Conclusion Serum level of visfatin can be used as an indicator to conjecture the severity degree of coronary artery lesion in patients with acute myocardial infarction.

Abstract:Aim To observe the effect of rosuvastatin on C-reaction protein（CRP）, interleukin-6 （IL-6）, brain natriuretic peptide （BNP）, left ventricular ejection fraction （LVEF）, left atrial diameter （LAD） and sinus rhythm maintenance in patients of the heart failure with paroxysmal atrial fibrillation. Methods All of 98 patients of chronic heart failure （CHF） with paroxysmal atrial fibrillation were converted to sinus rhythm after treatment. They were divided into rosuvastatin group （50 patinets） and control group （48 patinets） on the basis of routine CHF treatment. After follow-up 12 months, the change of CRP, IL-6, BNP, LVEF, LAD, the rate of recurrence of atrial fibrillation, incidence rate of heart failure were compared in two groups before and after treatment and the first and the twelfth month after discharge.

Abstract:Recent studies have shown that atherosclerosis is essentially a chronic vascular inflammatory disease, in which interleukin-1β plays a key regulatory role. Vascular endothelial cell damage caused by a variety of factors can lead to overexpression of interleukin-1β. By combination with interleukin-1 I receptor and activating NF-κB signaling pathway，interleukin-1β induces the synthesis of a large number of secondary inflammatory mediators, such as IL-6, TNFα. The results of animal experiment and clinical trials showed that antagonizing interleukin-1β signaling pathway was conducive to improving cardiac remodeling and reducing the risk of atherosclerotic cardiovascular events. The treatment of atherosclerosis targeting interleukin-1β has promising prospects for clinical application.

Abstract:Atherosclerosis （As） is a major cause of coronary artery disease （CAD）, and foam cells are a main reason of As, the accumulation of excess cholesterol in macrophages forms foam cells, so reducing the accumulation of cholesterol results in reducing the formation of foam cells, which may become an effective method for the treatment of As. ATP-binding cassette transporter A1 （ABCA1） mediates the transport of cellular cholesterol and phospholipids to ApoAⅠ to generate nascent HDL particles, which makes clearance of excess cholesterol from cells to the liver for excretion to the bile and feces, a process called reverse cholesterol transport （RCT）. ABCA1 can suppress inflammatory response and induce vascular endothelial cells changes, participate in oxidative stress, affect As by different metabolic pathways, and the effects of different genotypes on As is different. Therefore, ABCA1 plays an important role in the form and development of As.

Abstract:Inflammation mediated endothelial dysfunction plays an important role in the occurrence and development of obesity-induced cardiovascular disease. Omentin is a kind of anti-inflammatory adipokines, and there is a close relationship between obesity endothelial function. Regular exercise can produce anti-inflammatory effect, improve endothelial function in obese subjects and thereby reduce the risk of cardiovascular disease, through enhancing the expression and secretion of omentin in obesity. This paper is a review of recent research status of this field. It may provide a theoretical thinking for the further research of biological mechanisms of the regular exercise to improve the obesity-related vascular endothelial function.

Abstract:Berberine is a traditional Chinese herbal medicine. As a long term clinical antimicrobial drug, it has been used in treating bacterial diarrhea. In recent ten years, the domestic and foreign scholars have extensively studied on anti-atherosclerosis of berberine. In this paper, improvement of blood lipids of berberine and its mechanism of anti-atherosclerosis are reviewed from regulating blood plasma lipids, suppressing the formation of atheromatous lipid plaque on the arterial wall, and so on.