Abstract:Aim To observe the influence of Huayuqutan recipe on the metabolism of cholesterol in apolipoprotein E gene knockout （ApoE^－/－） mice, so as to illuminate the role and mechanism of Huayuqutan recipe on atherosclerosis. Methods Ten C57BL/6/J mice were used as blank group. Thirty ApoE^－/－ mice were randomly divided into model group, Huayuqutan recipe group and simvastatin group. The levels of serum lipid were tested by biochemical methods. HE and oil red staining method were used to observe the aortic wall injury of ApoE^－/－ mice atherosclerotic mice.RT-PCR and Western blot methods were used to test the changes of expression of low-density lipoprotein receptor （LDLR）, lecithin-cholesterol acyl transferase （LCAT） in liver and CD36 in aortic wall. The expression of CD36 in macrophages was detected by flow cytometer. Results Compared with the blank group, the serum triglyceride （TG）, total cholesterol （TC） and low-density lipoprotein cholesterol （LDLC） levels and the expression of CD36 in aortic wall and macrophages were significantly increased while high-density lipoprotein cholesterol （HDLC） level and expression of LDLR, LCAT in liver were significantly decreased in the model group. In Huayuqutan recipe group and simvastatin group, the serum con tent of TG, TC and LDLC were lower and HDLC was higher compared with the model group. At the same time, the expression of LDLR, LCAT in liver were up-regulated and CD36 in aortic wall as well as macrophages was obviously down-regulated. Conclusion Huayuqutan recipe could inhibit the formation of the atherosclerotic plaques, which might be related to the regulation of blood lipid, the expression of LDLR, LCAT as well as CD36.

Abstract:Aim To investigate the role of selective inhibition of the PI3K/Akt/mTOR signaling pathway on prompting rabbit primary macrophage autophagy. Methods Primary macrophages were obtained intraperitoneally from the New Zealand rabbits and then were co-cultured with the phosphatidylinositol 3-kinase （PI3K） inhibitor LY294002 （10 μmol/L）, protein kinase B （Akt） inhibitor triciribine （20 μmol/L）, mammalian target of rapamycin （mTOR） inhibitor rapamycin （10 nmol/L） and no drugs respectively for 12 hours. Ultrastructural changes of macrophages were examined by transmission electron microscopy. The expression level of microtubule-associated protein light chain 3Ⅱ（LC3Ⅱ） was assayed by immunofluorescence. Protein expression levels of Akt, mTOR, phosphorylation of protein kinase B （p-Akt）， phosphorylation of mammalian target of rapamycin （p-mTOR） and autophage related protein Beclin-1 and autophagy protein 5 and 12 conjugated form （Atg5-Atg12） were measured by Western blot. Mondansylcadaverine （MDC） staining was used to see autophagy lysosome changes. Results Compared with the control group, few autophagosomes and vacuoles were detected in group LY294002 while plenty of typical autophagosomes were detected in group rapamcin and triciribine. The expression levels of Beclin-1 and Atg5-Atg12 decreased significantly in group rapamcin, while increased significantly in group rapamcin and triciribine. The fluorescence microscope showed few dots of LC3Ⅱ in group LY294002 and many in group rapamcin and triciribine. The expression of p-Akt and p-mTOR increased obviously in group rapamcin while the former increased a lot and the latter decreased in group rapacin after co-culturing of 4 hours. The expression of p-mTOR decreased significantly in the treated groups, however, p-Akt decreased significantly in group rapamcin and triciribine but increased obviously in group rapamci after 12 hours’ co-culture. There were no significant differences on the total AKT and mTOR levels among the treated groups. MDC staining showed decreased autophagic lysosomes in group LY294002 and increased autophagic lysosomes in group rapamcin and triciribine. Conclusion Selective inhibition of PI3K/Akt/mTOR signaling pathway can promote rabbit primary macrophage autophagy while inhibition of PI3K suppress macrophage autophagy by other signaling pathway.

Abstract:Aim To investigate the influence of atorvastatin on the expression of phosphatase and tensin homolog deleted on chromosome （PTEN） involved in human CD4＋T lymphocytes in vitro. Methods Human CD4＋T cells obtained from healthy individuals were activated with phytohemagglutinin （PHA） and treated with atorvastatin. The levels of the inflammatory cytokines TNF-α， IL-6 and IL-10 were tested by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescent quantitative polymerase chain reaction （RFQ-PCR） was used to measure the expression of PTEN mRNA in CD4＋T cells, which were assessed at the protein level by Western blot as well. Results Compared to the blank group, the stimulation of PHA obviously increased the mRNA and protein expression of PTEN and those serum TNF-α and IL-6 secretion （P<0.05）, while the level of IL-10 was different between two groups （P>0.05）. Compared with those of PHA stimulation group, different concentration atorvastatin significantly increased the protein level of PTEN expression and serum IL-10 secretion （P<0.05）, while serum TNF-α and IL-6 production were reduced （P<0.05）. Serum TNF-α and IL-6 secretion were negatively related to the expression of PTEN through the linear related analysis （r＝－0.837 and r＝－0.816, P<0.01）, and IL-10 positive relation with the expression of PTEN （r＝0.753, P<0.05）. Conclusion The anti-inflammatory effects of atorvastatin are mediated by regulating the expression of PTEN in CD4＋T cells.

Abstract:Aim To investigate the effects of bradykinin （BK） of different concentrations on the survival, migration and apoptosis of human umbilical cord blood derived endothelial progenitor cells （hEPC） in vitro. Methods Total mononuclear cells （MNC） were isolated from human cord blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin- coated culture dished. After 7 days cultured, attached cells were isolated and assessed by immunofluorescence. Differentiating EPC were characterized as adherent cells double positive for both DiI-acLDL and FITC-UEA-1. Flow cytometry were used to further confirm EPC. EPC were treaed with BK of different concentrations （1, 10, 100 nmol/L）, or HOE140 （BK B2 receptor inhibitor） plus BK （10 nmol/L） for 16 h. The effects of BK on EPC’ survival, migration and apoptosis were assayed with MTT assay, Transwell chamber assay, flow cytometry assay using Annexin V-FITC/PI assay and Hoechst 33342 staining, respectively. Results BK at 1 and 10 nmol/L significantly improved the ability in the survival, migration and anti-apoptosis of hEPC （P< 0.05）, which was not at the concentration of 100 nmol/L （P> 0.05）. And the effects of BK on these functions of hEPCs could be blocked by HOE140 （P< 0.05）. Conclusion The present study established that BK in a certain concentration range could improve hEPC’ survival, migration and anti- apoptosis capability, and this effect might be mediated by BK-B2 receptor.

Abstract:Aim Using animal model to determine the effect of recombinant human tumor necrosis factor-like weak inducer of apoptosis （rhTWEAK） on aortic endothelial diastolic function and tissue factor and its inhibitor. Methods C57BL mice were randomly divided into four groups: （1）rhTWEAK group: given rhTWEAK by intraperitoneal injection （2）rhTWEAK＋anti-rhTWEAK group: after intraperitoneal injection with anti-TWEAK factor, then treated with rhTWEAK factor 15 minutes later （3）IgG group: treated with nonspecific IgG （4）Control group: given the same amount 0.3 mL of normal saline injection. After 7 weeks, we took eyeball blood to separate supernatant and thoracic aortic area to detect the vasodilatation function. Circulating endothelial nitric oxide synthase （eNOS）, high-sensitivity C-reactive protein （hs-CRP）, nitric oxide （NO） and tissue factor （TF） and tissue factor pathway inhibitor （TFPI） were detected in mouse plasma by enzyme linked immunosorbent serologic assay. Results （1）We found that endothelial diastolic function and the concentration of NO and eNOS in plasma in TWEAK group obviously decreased, TF levels increased significantly and the TFPI decreased when compared with control rhTWEAK＋anti-rhTWEAK and IgG group, those factors had significant difference （P<0.05）, however the hs-CRP content had no obvious change （P>0.05）. （2）Our results show that the endothelial diastolic function and the content of NO eNOS TF and TFPI and hs-CRP have no obvious difference in the control rhTWEAK＋anti-rhTWEAK and IgG group （P>0.05）. Conclusion rhTWEAK can induce endothelial dysfunction and blood coagulation factor content changes, and this may be a mechanism of vascular endothelial damage in early stage atherosclerosis.

Abstract:Aim To investigate the effect and mechanism of aspirin on human umbilical vein endothelial cells （HUVEC） senescence exposed to high glucose condition. Methods The cultured HUVEC were treated with concentrations of glucose at 5.5, 33 mmol/L and glucose at 33 mmol/L with aspirin at 0.01, 0.1, 1 and 3 mmol/L for 48 h. SA-β-gal staining was used to evaluate senescence. Telomerase activity was detected by PCR-ELISA. Cell cycle and the level of reactive oxygen species （ROS） were detected by flow cytometry. Results Endothelial cells exhibited the characteristic of senescence, increased the number of β-gal positive cells, decreased telomerase activity significantly, enhanced the proportion of HUVEC in the G0/G1 phase and reduced that in the S phase and increased the level of ROS after exposure to high glucose （33 mmol/L, P<0.05）. All these phenomena were reversed by 0.01, 0.1 and 1 mmol/L aspirin remarkably （P<0.05）. However, there was no significant difference between 3 mmol/L aspirin group and high glucose group. Conclusions 0.01, 0.1 and 1 mmol/L aspirin delay endothelial cell senescence exposed to high glucose. This positive effect may be associated with decreasing oxidative stress.

Abstract:Aim To compare the difference in blood pressure control rate between ambulatory blood pressure（ABP） and office blood pressure （OBP） in chronic kidney disease （CKD） patients. Methods We enrolled 225 CKD patients complicated with hypertension in our hospital during 2012.5～2013.2. The general and blood pressure information about these patients were recorded. Results （1） Compared with CKD 1～2 patients, the decline of blood pressure control rate of office systolic blood pressure, 24 h average blood pressure, especially the night time blood pressure in CKD 5 patients were significantly different （P<0.05）. （2）The blood pressure detection rate evaluated by OBP and ABP were different. In CKD 1～2 patients, ambulatory blood pressure detection rate were lower than office blood pressure control rate（79.6% vs 61.3%, P0.038）, however, in CKD 5 patients, the result was on the contrary（83.5% vs 93.0%, P0.029）. Conclusions （1） With the deterioration of renal function, the blood pressure control rate decreases. （2）There are differences between the blood pressure control rate evaluated by two blood pressure measurements in CKD patients of different clinical stage, so it is necessary for CKD patients to take ambulatory blood pressure measurements.

Abstract:Aim To elucidate the relationship between autophagy of peripheral blood monocyte （PBM） and the vulnerability of atherosclerotic plaques. Methods Forty patients with stable angina pectoris （SAP）, 40 patients with acute coronary syndrome （ACS） compromised the study groups. All patients underwent coronary angiography （CAG） and intravascular ultrasound （IVUS） examinations. The expression levels of autophagy related protein Beclin-1, microtubule-associated protein 1 light chain 3 （MAP1-LC3） and Atg5-Atg12 complex in PBM were detected by Western blot. MAP1-LC3 （autophagy-specific protein） in the PMB was also examined by laser scanning confocal microscope. Results Sixty two plaques were detected by IVUS in the SAP patients, which included 30 fibrous ones, while 71 plaques were evidenced in the ACS group, which encompassed 40 lipid ones. The expression levels of Beclin-1, MAP1-LC3 and Atg5-Atg12 complex in PBM in ACS patients were significantly lower than those in SAP patients （P<0.01）. Conclusion The patients in SAP group had more stable fibrous plaques than those in the ACS group （P<0.01）, while the ACS patients had more vulnerable soft lipid ones than those in the SAP group （P<0.01）. The autophagy of PBM in patients with coronary heart disease （CHD） decreases with the increasing vulnerability of atherosclerotic plaques. Enhancing the autophagy of PBM may be a potential therapeutic target of stabilizing atherosclerotic plaques, which will reduce the acute coronary events and lower the mortality of patients with CHD.

Abstract:Aim To explore the impact of the different baseline level of high-sensitivity C-reactive protein （hs-CRP） on the brachial-ankle pulse wave velocity （baPWV）. Methods Prospective cohort study method was used in our study. 101 510 workers who had participated in the 2006-2007 Kailuan health examination were stratified randomly, and 5 440 participants with sufficient information for questionnaires and blood biochemical tests were recruited. In the 2010-2011 Kailuan health examination the baPWV was tested, and 4 651 were included for the final analysis. According to the concentration of hs-CRP in 2006-2007 Kailuan health examination, the participants were divided into three groups, and multivariate linear regression analysis and multivariate Logistic regression analysis were used to calculate the rate ratios for baPWV. Results （1）The average baPWV in hs-CRP<1 mg/L group （n＝2 682）, 1 mg/L≤hs-CRP≤3 mg/L group （n＝1 307） and hs-CRP>3 mg/L group （n＝662） were respectively 1 505.82 cm/s, 1 612.48 cm/s, 1671.04cm/s （P<0.001） （2）After adjustment for other risk factors, with the every one unit increasing of baseline lg_hs-CRP, baPWV increased 6.448 cm/s in the multiple linear regression analysis （P＝0.034） （3）Compared with participants whose hs-CRP<1 mg/L, the RR for those with hs-CRP>3 mg/L was 1.34 （95%CI 1.04-1.72） in multivariate Logistic regression analysis. Conclusions The baPWV gradually increased with the increasing of baseline hs-CRP concentration. Serum hs-CRP>3 mg/L was an independent risk factor for increased baPWV in the general population, especially in men.

Abstract:Aim To investigate the correlation between the p53 gene codon 72 polymorphism and hyperlipidemia. Methods Blood samples were collected from 202 people in hyperlipidemia group（both the level of total cholesterol and low-density lipoproteins increased） and 194 people in control group. PCR-RFLP and PCR method was used to detect the genotypes of p53 codon 72 polymorphism. Results The genotype of the p53 gene codon 72 was ArgArg/ArgPro/ProPro, and the frequencies of genotype in hyperlipidemia group were 39.1 %, 45.0% and 15.8%, respectively.The frequencies of genotype in control group were 28.4%, 55.7% and 16.0%, and the difference of distribution of genotype was not significant（P>0.05）. The frequencies of genotype of women in two groups were 42.8%, 42.1%, 15.1% and 27.3%, 58.0%, 14.7%, respectively, and the difference was statistically significant（P0.012）. The genotype distribution of men in two groups was not significantly different. The differences of allele frequency distributions in the two groups and in men or in woman were not significant（P>0.05）. Conclusion There was a correlation between the genotype distribution of p53 gene codon 72 and hyperlipidemia. The risk of hyperlipidemia was increased in women who carried the Arg/Arg genotype. The genotype Arg/Arg may be a risk factor of hyperlipidemia.

Abstract:Aim To investigate the correlation of matrix metalloproteinase-9 （MMP-9） levels and MMP-9-1562C＞T polymorphism with acute ischemic stroke （IS） and its “Trial of Org 10172 in Acute Stroke Treatment” （TOAST） subtypes in Uygur nationality. Methods A total of 284 patients with acute IS were enrolled using the method of case-control study, based on the standard of new TOAST classification, 91 patients were atherothrombosis （AT）, 150 patients were small artery disease （SAD） and 43 patients were cardioembolism （CE）. Meanwhiie, 226 age-and sex-matched physically healthy subjects were used as control group. The levels of serum MMP-9 and-1562C＞T gene polymorphism in each group were measured and analyzed by enzyme-linked immunosorbent assay and restriction fragment length polymorphism. Results The serum MMP-9 level in the IS group was sigrfificantly higher than that in the control group （0.308±0.033 mg/L vs 0.087±0.011 mg/L, t＝7.813, P＝0.000）. The serum MMP-9 level in the AT group and CE group were significantly higher than those in the groups of SAD （0.350±0.030 mg/L vs 0.261±0.029 mg/L, t＝4.156, P＝0.001 0.317±0.043 mg/L vs 0.261±0.029 mg/L, t＝2.877, P＝0.031）. Multivariate Logistic regression analysis showed that the increased serum MMP-9 level was an independent risk factor for ischemic stroke （OR: 1.012, 95% CI: 1.007～1.016, P<0.001）. There was no significant difference in the frequencies of genotype （χ²＝3.558, P＝0.058） and allele （χ²＝3.567, P＝0.059） of MMP-9-1562C＞T between the IS group and the control group. However, there were significant difference in the frequencies of genotype （χ²＝5.097, P＝0.024） and allele （χ²＝5.439, P＝0.02） of MMP-9-1562C＞T between the AT group and the control group. The CT＋TT genotype frequency in the AT group was significantly higher than the control group （24.2% vs 13.7%）. Multivariate Logistic regression analysis showed that MMP-9-1562C＞T polymomhism was not an independent risk factor for the IS group （OR: 0.821, 95% CI: 0.622～1.059, P＝0.124）, but it was an independent risk factor for the AT group （OR: 1.768, 95% CI: 1.178～2.677, P＝0.007）. Conclusions The serum MMP-9 level increased in Uygur patients with IS, especially in the patients with AT.

Abstract:Aim To evaluate the predictive value of epicardial fat tissue （EFT） on coronary heat disease （CHD）. Methods Overall, 333 cases who underwent coronary angiography were classified into CHD groups and non-CHD group. EFT was measured by echocardiography. The obtained data was compared by receiver operating characteristic（ROC） curve and multi-logistic regression. Results The EFT in CHD group was significantly higher than that in non-CHD group （6.4±0.8 mm vs 4.8±0.7 mm, P<0.01）. ROC curve analysis showed EFT could reliably discriminate CHD patients （AUC: 0.834, 95% CI: 0.775～0.894, P<0.01）. Multivariate regression analysis also showed EFT was independent predictor of CHD ［OR5.17 （95% CI: 3.10～8.63）, P<0.01］. Conclusion This study showed EFT was independent predictor of CHD.

Abstract:Aim To investigate the application opportunity of intra-aortic balloon pump （IABP） for patients with acute myocardial infarction （AMI） complicated by pump failure and its influence on therapeutic efficacy for these patients.Methods 85 patients with acute myocardial infarction complicated by pump failure were randomly divided into two groups according to application opportunity of IABP. Immediate IABP group （43 cases） underwent the IABP surgery immediately after diagnosis. Necessary IABP group （42 cases） after positive drugs therapy, whose systolic blood pressure remains ＜90 mmHg, underwent IABP surgery. And all the patients underwent emergency percutaneous coronary intervention （PCI） as soon as possible. The difference of door-to-balloon time, survival rate of the 1 week and 3 month postoperative in two groups were compared, left ventricular ejection fraction （EF） of 1 day and 3 month were measured. Results Door-to-balloon time of necessary IABP group was significantly greater than the immediate IABP group （96.7±31.2 min vs 78.3±35.6 min, P<0.05）. The survival rate of necessary IABP group after 3 months was significantly lower than a week postoperative （57.1% vs 69.0%）, and lower than instantly IABP group （57.1% vs 67.4%）, there were significant differences （P<0.05）. EF of two groups after three months were continuously decreased, the necessary IABP group declined more than the immediate IABP group （7.6%±3.5% vs 4.2%±3.1%, P<0.05）. Conclusion Early use of IABP for patients with acute myocardial infarction complicated by pump failure, can shorten revascularization time, reduce mortality and protect left ventricular systolic function.

Abstract:Aim To evaluate the myocardial reperfusion of patients with acute myocardial infarction under treatment of fasudil by myocardial blush grades （MBG） after coronary stent implantation. Methods Ninty-two patients with acute myocardial infarction were randomly divided into treatment group and control group. Patients in the treatment group were given fasudil intra-vena twice a day for 7～10 days besides the ordinary treatment while the ones in the control group were only given ordinary treatment. Coronary artery angiography and coronary stent implantation were taken after treatment and myocardial reperfusion was identified by MBG. Results The MBG of treatment group improved significantly （P<0.05） compared with control group. Conclusions The routine treatment plus fasudil hydrochloride therapy can improve myocardial reperfusion in patients with acute myocardial infarction after coronary stent implantation.

Abstract:Aim To probe into clinical application value of off-pump coronary artery bypass grafting（OPCABG） in elderly patients with coronary heart disease. Methods Selected from January 2010 to December 2012, 308 elderly patients with coronary heart disease were treated with coronary artery bypass grafting （CABG） in our department. According to the surgical approach, the patients were divided into on-pump CABG group （32 cases） and off-pump CABG group （276 cases）. The therapeutic effects of the patients with CABG in two groups were compared and analysed. Results Compared with on-pump CABG group, the mortality in 30 days, number of bypass vessels, rate of reoperation, incidence of acute renal failure, respiratory failure and new onset atrial fibrillation, blood transfusion, incidence of cerebral infarction and pulmonary infection were significantly reduced in off-pump CABG group, and ventilator time, intubation time and stayying-inhospital time were significantly shorter, which were statistically significant （P<0.05）. Conclusions Off-pump CABG could improve the effect effectively， reduce the mortality and the incidence of complications for the elderly patients with coronary heart disease. It could be used as the preferred method of surgical treatment for elderly patients with coronary heart disease and promote the further application.

Abstract:Aim To investigate the influence on the level of intracellular reactive oxygen species （ROS） by staining the cell nuclei using two fluorescent dyes —Hoechst 33342 and DAPI, respectively. Methods Vascular smooth muscle cells （VSMC） were stimulated with advanced glycation end products （AGE） for 10 minutes and then incubated with 2’,7’-dichlorofluorescin diacetate （H2DCFDA）. After that, cell nuclei were stained with Hoechst 33342 and DAPI respectively. And through the analysis of the number of labeled nuclei and the level of intracellular ROS by fluorescence microscopy, the fluorescence intensity of intracellular ROS were detected by staining with two different fluorescent dyes.Results After staining with Hoechst 33342 for 5 min, cell nuclei were labeled immediately and the number of them did not change with the increase of staining time. However, there were only a few cell nuclei could be labeled when the cells were stained with DAPI for 5 min, with the increase of staining time, more and more cell nuclei could be labeled. Surprisingly, the fluorescence intensity of Hoechst 33342 group showed no significant differences staining at 5, 10 and 20 min.

Abstract:Atherosclerosis causes severe cardiovascular diseases, and those diseases are the most serious threats to public health. Inflammation was generally considered as a novel risk factor of atherosclerosis besides some classic factors, such as hyperlipemia. The Nod-like receptor protein-3 （NLRP3） inflammasome is a large multimeric danger-sensing platform, as one of innate immunity response, promotes the activation of Caspase-1 and mediates the mature of pro-inflammatory cytokines interleukin-1β （IL-1β） and IL-18. Evidences have been provided that NLRP3 inflammasome has a critical role in the development of atherosclerosis. Here we discuss the advance in the role of NLRP3 Inflammasome in atherogenesis, and the potential targets of atherosclerosis according to NLRP3 inflammasome.

Abstract:With the improvement of people's living standards and diet, the incidences of hyperuricemia and artery atherosclerosis increase. Research revealed that the two diseases were closely related. But the mechanism on how hyperuricemia leads to atherosclerosis is not very clear yet. Hyperuricemia may facilitate the progress of atherosclerosis through these ways: the direct effect of soluble uric acid, urate crystals inflammatory reaction, and hypercholesterolemia which enhances the correlation between high uric acid and atherosclerosis. Reduction of blood uric acid could reduce the occurrence of atherosclerosis and improve the survival rate of patients with cardiovascular and cerebrovascular diseases. The improvement of the quality of long-term survival of patients undoubtedly has important clinical significance. Therefore, in-depth and extensive reseaches are necessary in elucidating the underlying mechanisms.

Abstract:Lipoprotein associated phospholipase A2 （Lp-PLA2） is an enzyme produced by inflammatory cells and attached to low density lipoprotein （LDL）. Lp-PLA2 hydrolyzes the oxidized LDL and releases inflammatory meditor to promote atherosclerosis. There is a significant association between the single nucleotide polymorphisms in the PLA2G7 gene and coronary heart disease. Lp-PLA2 is a novel inflammatory biomarker and has close relationship with the propensity of atherosclerosis. Lp-PLA2 has received attention gradually as an independent risk predictor and may be a new therapeutic target of coronary heart disease. This paper reviewed the recent research in the correlation between Lp-PLA2 and coronary heart disease.