Abstract:The major cause of human and experimental heart failure is related to the deregulated sarcoplasmic reticulum （SR） calcium cycling, controlled by several protein complexes. Those complexes, including protein kinase, protein phosphatase and some interacting proteins or subunits, work together to maintain normal cytosolic calcium homeostasis. Among them, SR calcium transport is regulated by SR calcium ATPase and the phosphorylated protein: phospholamban （PLN）. Recently, some other proteins，such as: protein phosphatase 1 inhibitor 1, heat shock protein 20 （HSP20） and HS-associated protein X-1 （HAX-1） have been reported to play an important role in SR calcium handing as well as cardiac function. This review will focus on the recent findings in these PLN interacting complex and their contribution in cardiac pump function as well as the potential clinical application.

Abstract:Aim To observe the effects and mechanism of glycyrrhiza acid （GA） on lipid metabolism and development of atherosclerotic lesions in apolipoprotein E gene knockout （ApoE^－/－） mice. Methods Eighteen female ApoE^－/－ mice were randomly divided into control group and glycyrrhiza acid treatment group. All mice were fed with Western diet for 12 weeks, glycyrrhiza acid 100 mg/（kg·d） was intragastrically given to hyperlipidemic mice and 0.9% NaCl as the control. The weight of mice was detected per 4 weeks, the levels of plasma lipids and glucose were determined enzymatically, serum paraoxonase 1 （PON1） activity was measured by using paraoxon as the substrate. The extent of aortic atherosclerosis was examined both in oil red O-stained cross-sections of the proximal aorta （8 m cryosections） and by en face analysis with quantitation using Image J System. The expression levels of gene in liver were detected by real-time PCR and Western blot. Results The average weight of mice was increased and dyslipidemia was aggravated during being fed with Western diet. The increasing level of mice weight, the level of plasma triglycerides and glucose had no obvious change between both groups. However, the level of total cholesterol was statistically decreased （P<0.05） in ApoE^－/－ mice with glycyrrhiza acid treatment compared with control group. In mice of glycyrrhiza acid group, the atherosclerotic lesions of aortic root and aorta were reduced by 21% to 22% versus control mice. Glycyrrhiza acid obviously up-regulated the mRNA level of scavenger receptor BⅠ （SR-BⅠ） and ATP-binding cassette transporter A1 （ABCA1） in mouse liver. Meanwhile, glycyrrhiza acid significantly increased the serum antioxidative enzymes PON1 activity by up-regulation of the PON1 expression level （P<0.01）, and the expression level of cellular methionine sulfoxide reductase A （MsrA） was also increased. Conclusions Glycyrrhiza acid inhibits the development of the atherosclerosis, the mechanism might be related to the regulation of cholesterol metabolism and improvement of antioxidation.

Abstract:Aim To study hypolipidemic and anti-atherosclerosis effects and mechanisms of grape-seed polyphenols （GSP） in LDLR^-/- mice. Methods 7～8 weeks male LDLR^-/- mice （n32） were randomly divided into 4 groups with 8 mice in each group. One group was orally administrated with saline and fed with chow diet （normal control）. The other three groups were fed with a high fat diet containing 0.5% cholesterol and 20% lard and orally administrated with saline （hyperlipidemia control）, 5 mg/（kg·d） ezetimibe （positive drug control）, and 750 mg/（kg·d） GSP, respectively. Ezetimibe and GSP were dissolved in saline for administration. After a 12-week regimen, blood samples of the mice were collected by retro-orbital blood drawing for determining plasma TC and TG levels. All mice were then sacrificed and mouse aortas were isolated and dissected under microscope, and atherosclerotic lesions on aortic walls were stained by oil red O. Mouse aortic roots were frozenly sectioned for HE and oil red O staining, and Mac-2 immunohistochemistry staining as well. Besides, gene expression in mouse livers, related to lipid synthesis, oxidation, and metabolism, were measured by RT-PCR. Results Compared to saline treated hyperlipidemia control group, treating mice with GSP significantly decreased their TC and TG levels, significantly reduced atherosclerosis lesion area in mouse aortas and aortic roots, decreased macrophage infiltration in plaques, and up-regulated gene expression of LRP1 and CYP27A1 in mouse livers. Conclusions GSP has hypolipidemic effects and ameliorates atherosclerosis in LDLR^-/- mice. The hypolipidemic mechanisms of GSP may relate to regulation of alternative and neutral synthetic pathways of cholesterol metabolism, which results in an increase of cholesterol transference into bile acid, and improvement of lipid metabolism. The anti-atherosclerosis effects of GSP can be concluded to decrease of plasma TC and TG levels and reduction of foam cell formation.

Abstract:Aim To investigate the effect of heat shock protein 65 （HSP65） as the important inflammatory substances in atherosclerosis on cellular immune function and properties of high density lipoprotein in apoptotic human T cell lymphoma Jurkat cells in vitro and explore the possible molecular mechanisms of immune system and lipid metabolism. Methods Jurkat cells activated by 5 mg/L concanavalin A （ConA） were treated with different concentrations of HSP65, and the cell proliferation were detected by CCK-8 assay. The concentration of cytokines interferon-γ （IFN-γ） and interleukin-10 （IL-10） were measured by ELISA. Cholesterol efflux was assayed by liquid scintillation spectrometry. Expression of ABCA1, ABCG1, SR-BⅠ, PPARγ and LXRα mRNA were detected by RT-PCR, respectively. Results ConA enhanced the activity of Jurkat cells after 48 h incubation. HSP65 increased the proliferation activity of Jurkat cells gradually （P<0.001）. The cholesterol efflux and IL-10 level were significantly lower, whereas IFN-γ level elevated in 0.5 mg/L group and 1 mg/L group than those in control group （P<0.01）. Compared with control group, the treatment of Jurkat cells with 0.5 mg/L and 1 mg/L HSP65 resulted in a concentration-dependent decrease of ABCG1, ABCG1, SR-BⅠ, PPARγ, LXRα mRNA expression （P<0.01）. Conclusions HSP65 within the concentration range of 0.5～1 mg/L can promote the proliferation and intensify the cellular immune response, which may attenuate cholesterol efflux by down-regulating ABCG1, ABCG1, SR-BⅠ, PPARγ, LXRα mRNA expression in T cells.

Abstract:Aim To develop a rabbit model of vulnerable atherosclerotic plaque by cryogenic gas-induced endothelial injury combined with high-fat diet. Methods 24 healthy male New Zealand rabbits （3 months） were randomly divided into three groups: control blank group （group A）, sham-operated group （group B） and temperature control gas injury group （group C） with 8 in each group. After one week high-fat diet, the rabbits in group C were underwent carotid artery intima injury using cryogenic gas. The rabbits in group B suffered surgical operations without cryogenic gas. After the operation, the rabbits in group B and group C were kept feeding on high-fat diet for 12 weeks, whereas the ones in group A were kept on normal diet. All rabbits were killed after 13 weeks. Then paraffin-embedded and frozen sections were used for HE staining, Masson trichrome staining and elastic fiber staining and oil red O staining to analyze the changes of vascular morphology. Additionally, rat anti-rabbit macrophage antibody 11 （RAM11） monoclonal antibody was applied for animal immunohistochemistry staining. Results The rabbit carotid atherosclerosis established in our study was in line with the progress characteristics of vulnerable plaque, involving thin fibrous cap, large lipid core and macrophages and foam cells in the intima as well as endothelial cells shedding or local small scattered thrombosis. The rabbit carotid in sham-operated group showed intimal hyperplasia and partly showed small primary plaques infiltrated by macrophages, which was in line with the change of fatty streaks phase. There is no plaque visible in the control blank group. Conclusion It is viable and easy to develop a rabbit model of typical vulnerable atherosclerotic plaque by injuring carotid intima with temperature control gas combined with high-fat diet.

Abstract:Aim To observe the effect and mechanism of soluble epoxide hydrolase inhibitor t-AUCB regulating murine endothelial progenitor cells’ function. Methods Separated by density gradient centrifugation to obtain mouse bone marrow progenitor cells, different concentrations of t-AUCB and peroxisome proliferator activated receptorγ （PPARγ） blocker GW9662 pre-intervented endothelial progenitor cells, after that endothelial progenitor cell proliferation, adhesion, migration, angiogenesis ability were detected in vitro. Results From 0～100 μmol/L, as the concentration increases, t-AUCB can increase endothelial progenitor cell proliferation, adhesion, migration, angiogenesis ability in vitro. Conclusion Soluble epoxide hydrolase inhibitor t-AUCB involved in forward regulating endothelial progenitor cell’ function, and its function is associated with activation of PPARγ on endothelial progenitor cell.

Abstract:Aim To investigate whether acylated ghrelin can protect 3T3-L1 mouse adipocytes from inflammatory injury mediated by inflammatoral factor tumour-necrosis factor-α （TNF-α）. Methods Four experimental groups were set up: control group, TNF-α treated group, acylated ghrelin treated group and a group pretreated by acylated ghrelin followed by TNF-α treatment. After the completion of the intervention, the mRNA and protein levels of Toll-like recepter 4 （TLR-4）, nuclear factor κB p65 （NF-κB p65） protein phosphorylation level, the concentration of monocyte chemotactic protein 1 （MCP-1） and adiponectin in the supernatant were detected. Results ① Compared with control group, mRNA and protein expression level of TLR-4 were increased by TNF-α intervention, as well as level of NF-κB p65 protein phosphorylation and MCP-1 protein in the murine 3T3-L1 adipocytes （P<0.05,n5）. On the contrary, the adiponectin protein was decreased（P<0.05,n5）. ②Compared with control group, the level of TLR-4 mRNA and protein expression, as well as NF-κB p65 protein phosphorylation were significantly decreased in acylated ghrelin treated group （P<0.05,n5）. The adiponectin level in the cell supernatant was decreased （at 15 pmol/L most obvious） （P<0.05,n5 ）, while level of the MCP-1 went down without statistically significant difference （P>0.05,n5）. ③Compared with TNF-α（100 μg/L） treated group, the incubation of acylated ghrelin could antagonise TNF-α-induced activation of TLR-4/NF-κB pathway in mouse 3T3-L1 adipocytes, mRNA and protein expression levels of TLR-4, the level of NF-κB p65 protein phosphorylation were decreased （P<0.05,n5） in a dose-dependent manner. The secretion of MCP-1 and adiponectin did not change significantly in 3T3-L1 adipocytes （P>0.05,n5）. Conclusions TNF-α can activate TLR-4/NF-κB inflammatory pathways and increase secretion of pro-inflammatory cytokines （MCP-1）, while decrease the secretion of anti-inflammatory cytokines （adiponectin） in 3T3-L1 adipocytes. Acylated ghrelin can antagonise TNF-α-induced activation of TLR-4/NF-κB pathway in a dose-dependent manner in mouse 3T3-L1 adipocytes, but can not completely improve the secretion disorder of MCP-1 and adiponectin.

Abstract:Aim To investigate the relationship between the concentration of plasma lipocalin-2 （LCN2） and common carotid artery intima-medial thickness （CCA-IMT） in newly diagnosed type 2 diabetic patients. Method 166 newly diagnosed type 2 diabetic patients （age 35～70, duration≤1 year） and 65 healthy people were recruited. CCA-IMT was detected by high-resolution ultrasonography, plasma concentration of LCN2 was measured by enzyme-linked immunosorbent assay. Results Adjusting for age and sex, type 2 diabetics’ plasma concentration of LCN2 was significantly higher than that in the healthy group （P<0.05）. According to the three tilter of plasma concentration of LCN2, the highest concentration group’s CCA-IMT was significantly higher than that in the lowest concentration group （P0.026）.

Abstract:Aim To develop an approach based on targeted gene capture and next-generation sequencing to determine the genetic defects in patients with abdominal aortic aneurysm precisely and effectively and confirm the role of fibrillin-1 （FBN1） in the pathogenesis of abdominal aortic aneurysm. Methods We analyzed four abdominal aortic aneurysm patients. Peripheral blood was collected and genomic DNA was isolated. FBN1 were selected by a gene capture strategy, using GenCap custom enrichment kit. The enrichment libraries were sequenced on Illumina HiSeq 2000 sequencer for paired read 90 bp to determine the mutation frequency in FBN1. Variants which have been reported in dbSNP137 or 1000 genome and in-house database were filtered. Selected variants were further validated by PCR and Sanger sequencing. Results For the samples subjected to targeted next-generation sequencing, the average sequencing depths on the targeted regions were yielded from 448.15 to 536.61. Meanwhile, coverage of targeted exons were ranged from 99.5% to 99.7%. We identified one missense mutation in FBN1: c.2753 C＞G （p.Pro918Arg）, which was validated by Sanger sequencing. Conclusions Our results confirm the role of FBN1 in the pathogenesis of abdominal aortic aneurysm and demonstrated the robustness of targeted next-generation sequencing to precisely and rapidly determine genetic defects.

Abstract:Aim To investigate the relationship of serum levels of pregnancy-associated plasma protein-A （PAPP-A） with carotid atherosclerotic plaque in acute cerebral infarction. Methods Carotid artery intima-media thickness （IMT） and the characteristic of atherosclerotic plaque were examined by carotid ultrasonography and the levels of serum PAPP-A were measured by enzyme immunoassay in 79 patients with acute cerebral infarction and 64 normal controls.

Abstract:Aim To determine the platelet surface expression of glycoprotein Ⅱb/Ⅲa fibrinogen receptor （PAC-1） and P-selectin （CD62P） in diabetes mellitus （DM） patients with acute coronary syndrome （ACS） and to explore the relationship between them and plasma homocysteine （Hcy） level. Methods The platelet surface expression of PAC-1 and CD62P were measured by flow cytometry in 94 patients, including 40 ACS patients, 24 DM with ACS patients and 30 patients in control group. The correlation between platelet PAC-1, CD62P and plasma level of Hcy were investigated.

Abstract:Aim To compare the long-term clinical efficacy and safety of hybrid surgery and PCI in the treatment of coronary heart disease in patients with multivessel disease. Methods A prospective randomized controlled study was adopted from 2012 January to 2014 June, 102 cases in our hospital, in the diagnosis of coronary angiography for multivessel coronary artery disease, randomly divided into hybrid group （n53） and PCI group （n49）, patients accepted hybrid surgery in hybrid group, patients with multi branch coronary artery lesions accepted pure PCI treatment in PCI group. 2-years follow-up was after 1, 3, 6, 9 and 12 month. After 12 months, coronary angiography was adopted to evaluate target vessel patency rate and SYNTAX score. Clinical status of patients, and the primary and secondary end points, the rate of long term survival and cardiovascular adverse events were recorded. Results The amount of contrast agent of hybrid group was lower than that of PCI group （P<0.05）. There was significantly difference of average hospitalization days, total stent length, postoperative hs-CRP peak of the two groups （P<0.05）. The hospital adverse events of contrast-induced nephropathy, acute heart failure, recurrent angina pectoris and postoperative hypotension were significant different in the two groups （P<0.05）. There was no significant difference of repeat myocardial infarction, target vessel revascularization, cerebrovascular accident and death in the two groups （P>0.05）. All patients were followed for a median of 2.4 years follow-up, with an average of 16.2 ± 11.3 months. Between the two groups, there was remarkable difference of repeat myocardial infarction, target vessel revascularization, acute heart failure, recurrent angina pectoris （1.9% vs. 8.2%, 1.9% vs. 8.2%, 3.8% vs. 12.2%, 5.7% vs. 14.3%, P0.023）. 2-years total mortality was 3.8% in hybrid group, and 4.1% in PCI group. In the two groups, 1-year target vessel patency rate and SYNTAX scores were statistically significant （P<0.05）. Conclusions Compared with PCI treatment, coronary hybrid surgery can reduce adverse events in patients with multivessel disease in the long-term, and operation safety was high.

Abstract:Aim To explore the relationship between coronary artery disease（CAD） and the level of plasma Klotho protein in the aged. Methods A total of 138 patients over 60 years old with CAD （ experiment group） were enrolled， including 65 cases of single vessel lesion，73 cases of multivessels lesion. And a total of 140 controls above 60 years old without CAD were chosen. The levels of plasma Klotho protein were detected and compared between the two groups. The degree of coronary artery stenosis were assessed by Gensini scores. The correlation between coronary artery stenosis degree and the level of plasma Klotho protein was analyzed simultaneously. Results The level of plasma Klotho protein in experiment group was lower than that in control group. Moreover, the level of plasma Klotho protein in multivessels lesion group was lower than that in single vessel lesion group. The coronary artery stenosis degree was negatively correlated with the level of plasma Klotho protien. Conclusion The level of plasma Klotho protein may reflect the degree of coronary stenosis and could be used as an index to estimate the coronary artery stenosis degree.

Abstract:Aim To investigate the relationship between severity of coronary heart disease （CHD） and plasma level of high-mobility group box1 protein（HMGB1）, high sensitive C-reactive protein（hs-CRP ）. Methods The study enrolled 100 patients suspected of CHD. All patients underwent coronary artery angiography. The patients were divided into CHD group and control group. The CHD group was divided into several subgroups according to the type of disease, Gensini score and the number of diseased coronary arteries. Serum HMGB1, hs-CRP, renal/hepatic function, blood lipids, blood glucose, myocardial markers were measured before coronary artery angiography, and history of smoking, hypertension, diabetes mellitus were also asked. The plasma level of HMGB1 and hs-CRP were compared among all groups to explore the correlation between plasma level of HMGB1,hs-CRP and severity of CHD. All data were analyzed by SPSS 20.0 for windows. Results ①Serum HMGB1 and hs-CRP level of CHD group were higher than the control group（P<0.01）. Serum HMGB1 and hs-CRP level of AMI group were higher than the UAP group（P<0.05）.

Abstract:Aim To explore the relevant factors of the ischemic stroke patients with cerebral microbleeds, and to provide reference for clinical diagnosis. Methods 142 ischemic stroke patients were collected from the second hospital of Lanzhou university with taking examination of MRI from March 2013 till 2014. Clinical data and the MRI examination results of all the patients were recorded and all the patients were divided into the group of ischemic stroke with cerebral microbleeds and the group of simple ischemic stroke, according to the results of SWI. Possible relevant factors of cerebral microbleeds about ischemic stroke patients were analyzed by logistic regression analysis. Two groups of patients were analyzed by Mann-Whitney U test to explore the relation between CMB and leukoaraiosis. Results Ischemic stroke with cerebral microbleeds was associated with hypertension and leukoaraiosis（P<0.05）. It was not associated with sex, age, diabetes mellitus, hyperlipidemia and the history of taking anti-coagulants or anti-platelet drugs（P>0.05）. Compared with the group of simple ischemic stroke, the degree of leukoaraiosis was more serious in the group of ischemic stroke with cerebral microbleeds （P<0.05）. Conclusion Hypertension and leukoaraiosis were the relevant factors of ischemic stroke with cerebral microbleeds. Among ischemic stroke patients, the degree of leukoaraiosis in patients with cerebral microbleeds was more serious.

Abstract:Aim To investigate the correlation between myeloperoxidase（MPO） and coronary slow flow（CSF）. Methods Through corrected TIMI frame count（CTFC）, the randomly selected 600 patients without coronary artery stenosis by coronary angiography （CAG） were screened out of 52 patients of CSF as the study group, the other randomly selected 52 patients without CSF as the control group. Plasma levels of MPO and other blood biochemical index were obtained in all participants, and analysis of the difference between the two groups, logistic regression analysis was used to screen the risk factors of CSF. Results Baseline values were similar in two groups MPO levels of the CSF group were significantly higher than control group（93.70±37.75 μg/L vs 26.76±6.20 μg/L, P<0.001）, the difference was statistically significant Logistic regression analysis showed that high plasma MPO level was related to CSF（OR1.23,P0.001）.

Abstract:Aim To detect the distribution of cervicocerebral arteriostenosis and evaluate its risk factors. Methods A total of 131 patients with intracranial and extracranial arteriostenosis were examined with transcranial Doppler ultrasonography （TCD）. Potential risk factors for vascular disease in each patient were evaluated based on their medical history. Then the effects of related factors on arteriostenosis were probed. Results Intracranial arteriostenosis was found in 114 out of 131 patients（85.5%）, which was significantly higher than extracranial artery stenosis or occlusion（26/131, 19.8%, χ²118.790, P0.000）. The middle cerebral artery was affected in 84 patients（64.2%）, which is the most frequently affected artery, accounting for 52.7% in all stenotic arteries. Binary logistic regression analysis showed that hypertension, diabetes and smoking were independent factors significantly associated with cervicocerebral arteriostenosis. Conclusions Hypertension, diabetes and smoking were independent factors significantly associated with cervicocerebral arteriostenosis. Transcranial Doppler ultrasonography should be performed in these patients routinely.

Abstract:Aim To investigate the relationship between rate-pressure product and brachial-ankle pulse wave velocity （BaPWV） in middle-aged and older women. Methods A total of 5852 participants was selected with stratified random sampling from the 101510 workers of Tangshan Kailuan company who received the health examination. Among them 2043 women with integral data were recruited into the survey. The observation population was divided into four groups according to the rate-pressure product collected during 2010～2011 health examinations: quartile 1 （the rate-pressure product＜7500.00）, quartile 2 （7500.00≤the rate-pressure product＜8624.00）, quartile 3 （8624.00≤the rate-pressure product＜10121.00）, quartile 4 （the rate-pressure product≥10121.00）. Multivariate logistic regression analysis was used to analyze the the influence of the rate-pressure product on BaPWV. Results （1） There were 2043 women with an average age of 52.2±9.7 years old. Their mean of BaPWV was 1488.9 cm/s. （2）The mean of BaPWV for the above quartiles of the rate-pressure product were 1266.0, 1377.3, 1525.6, 1786.7 cm/s, respectively and the detection rates of BaPWV≥1400 cm/s were 19.5%, 37.2%, 59.6%, 84.3%, respectively. （3） Multiple logistic regression analysis showed that after adjusting for age and other risk factors, compared with quartile 1, the risk of increasing BaPWV was increased in quartile 2, 3 and 4 of the rate-pressure product and the OR values were 1.66 （95%CI 1.16～2.37）, 3.18 （95%CI 2.19～4.62） and 7.30 （95%CI 4.67～11.42）, respectively. Conclusion The rate-pressure product is positively associated with increasing BaPWV in middle-aged and older women.

Abstract:Diabetic macroangiopathy is the leading cause of death in diabetic patients. The remarkable pathological change of diabetic macroangiopathy is atherosclerosis. Diabetes mellitus can lead to premature and accelerated atherosclerosis.

Abstract:Hydrogen sulfide （H₂S） is the new gasotransmitter to regulate the function of cardiovascular system after nitric oxide （NO） and carbon monoxide （CO）. H₂S can hinder the development of atherosclerosis by protecting vascular endothelium and inhibiting vascular smooth muscle cell proliferation and foam cell formation. This protective effect of H₂S is achieved through the mechanisms such as anti-inflammation, against oxidative stress. Therefore, hydrogen sulfide is expected as a promising therapeutic target for atherosclerosis.

Abstract:Osteoporosis and atherosclerosis are both widely prevalent in ageing population, and induce serious morbidities and death. There is growing evidence that osteoporosis and atherosclerosis have clinical interrelation. Experimental studies indicate that bone and arterial wall share some matrix proteins and factors, such as osteoprotegerin, osteopontin, matrix Gla protein, osteocalcin and low density lipoprotein receptor-related protein. And both diseases may have the same physiopathologic mechanism. The osteoprotegerin/receptor activator of nuclear factor-κB ligand/receptor activator of nuclear factor-κB system, oxidized lipids, inflammatory and calcium metabolic abnormalities may participate in or accelerate these diseases. And drugs such as stains, bisphosphonates, thiazolidinediones and osteoprotegerin which based on the biological linkage of above entities may benefit both bone and vessel. This article reviews their clinical interrelations, matrix proteins and factors shared by bone and arterial wall, the same physiopathologic mechanism, and therapies based on their interrelation.

Abstract:Statins play an increasing important role in the primary and secondary prevention of cardiovascular disease, since hypercholesteremia has been proven to be the major risk factor for cardiovascular disease. It is one of the most common drug on prescriptions, behind antibiotics. However, with the increasing clinical use, other adverse reactions were found besides on the muscle, liver and kidney. American FDA announces diabetes risk in labeling for some cholesterol-lowering drugs at February 2012. The same year in December, China's FDA made a similar risk warning. Now, new-onset diabetes risk has become one of the most concern about adverse reactions. This review focused on the latest research about new-onset diabetes risk of statins.