Abstract:Vascular endothelial cells, macrophages, and smooth muscle cells (SMC) participate in atherogenesis.Recent studies show that as many as 70% of all cells in atherosclerotic lesions are smooth muscle cells-derived. Inflammatory cytokines and immune modulators secreted by smooth muscle cells in the atherosclerotic plaque promote smooth muscle cell proliferation, migration and inflammatory response, and activate and recruit macrophages to the atherosclerotic lesions in a autocrine or paracrine manner. Moreover, smooth muscle cells in atherosclerotic lesions express receptors for lipid and can take up modified lipoproteins, leading to a massive accumulation of cholesterol esters and the formation of foam cells in the atherosclerotic plaque. Thus, smooth muscle cells play a key role in the formation of atherosclerotic plaque, the role and mechanism of action of smooth muscle cells in the atherosclerotic plaque pathogenesis need to be further studied.

Abstract:Reverse cholesterol transport (RCT) is a process that promotes the efflux of cholesterol from the cells and transports it to the liver for metabolism. Its dysfunction plays a key role in the development of atherosclerosis (As). Chronic metabolic inflammatory diseases including diabetes, obesity promote the progress of As, in which pro-inflammatory cytokines and adipokines are important regulatory mechanisms for the regulation of RCT in vivo. The research papers and review collected in this issue studied the effect of nuclear factor κB (NF-κB), glycosylation end products Nε-carboxymethyllysine, adipokines Visfatin, etc. on the cholesterol efflux and the expression of key proteins such as adenosine triphosphate binding cassette transporter A1, Sortilin, and acyl-coenzyme A:cholesterol acyltransferase (ACAT) in RCT, which would clarify the molecular mechanism of chronic metabolic inflammatory diseases regulating RCT and cardiovascular disease development from different perspectives.

Abstract:Aim To provide an excellent animal model for the study of atherosclerosis, it generated apolipoprotein E gene knockout (ApoE－/－) rabbit. Methods ApoE gene of New Zealand rabbits were edited and F0 ApoE－/－ rabbits were created by applying CRISPR/Cas9 system. The efficiency and effect of ApoE gene targeted knockout was determined through PCR, TA cloning, Sanger sequencing, Western blot and lipid assay. ApoE＋/－ rabbits were obtained through transmitting by germline. Results Sanger sequencing results showed that ApoE gene was successfully targeted knockout. Serum lipoprotein content of ApoE－/－ rabbits increased in normal diet compared with wild type rabbits and can be transmitted by germline. Conclusion The successful establishment of ApoE－/－ rabbit provided a good animal model for the further study of atherosclerosis and other related diseases.

Abstract:Aim To explore whether nuclear factor kappa B (NF-κB) manipulates Sortilin expression to regulate lipid metabolisms and foam cell formation of lipid-laden THP-1 macrophage. Methods The levels of Sortilin mRNA and protein were detected by quantitative real-time PCR (qRT-PCR) and Western blot in lipid-laden THP-1 macrophage incubated with NF-κB activator phorbol-12-myristate-13-acetate (PMA) or its specific inhibitor ammonium pyrrolidinedithiocarbamate (PDTC). Under the lipid-laden THP-1 macrophage treated with Sortilin shRNA and together with PMA, cholesterol efflux from macrophage was measured by liquid scintillation counting apparatus, intracellular lipid contents were detected by high performance liquid chromatography, and the intracellular lipid droplets were stained with oil red O. Results PMA treatment increased the levels of Sortilin mRNA and protein in lipid-laden THP-1 macrophage, whereas PDTC incubation decreased macrophage Sortilin expression. When lipid-laden THP-1 macrophage was treated with PMA alone, the cholesterol efflux of macrophages was reduced, the intracellular lipid accumulation was increased, and foam cell formation was increased. Reversely, the cholesterol efflux of lipid-laden macrophage was increased, the intracellular lipid accumulation was decreased, and foam cell formation was reduced under the treatment with Sortilin shRNA and supplemented with PMA. Conclusion NF-κB promotes Sortilin expression to inhibit the cholesterol efflux from lipid-laden macrophage and accelerates the accumulation of intracellular lipid and the formation of foam cell.

Abstract:Aim To study the effect and mechanism of Nε-carboxymethyllysine (CML) on vascular smooth muscle cells (VSMC) foam cells formation. Methods Primary VSMC were extracted from the aorta of C57BL/6J mice using tissue affixing method and identified; Primary smooth muscle cells of 3～9 passages were divided into control group, oxidized low-density lipoprotein (ox-LDL) (50 mg/L) group, 1,0, and 100 μmol/L concentrations of CML and ox-LDL co-stimulation group. The levels of total cholesterol (TC), free cholesterol (FC) and cholesteryl esters (CE) were determined by a cholesterol content assay kit; the primary VSMC were divided into four groups:control group, ox-LDL-induced model group, and ox-LDL co-stimulated with CML group, RAGE siRNA silencing group. Western blot and qRT-PCR was used to detect receptor for advanced glycation end products (RAGE) and cholesterol efflux end regulator ATP binding cassette transporter A1 (ABCA1); The degree of foaming of smooth muscle cells in each group was detected by oil red O staining and oil red extraction experiments. Results Compared with control group, CML significantly increased smooth muscle intracellular TC, CE and FC levels; Cholesterol efflux experiments showed that cholesterol efflux rate decreased with increasing CML concentration; Western blot showed that compared with the control group, RAGE in the CML group was significantly higher, ABCA1 was significantly lower, and RAGE siRNA decreased expression of RAGE, and increased ABCA1 levels. Oil red staining showed that ox-LDL could promote lipid accumulation in smooth muscle cells, and CML could enhance its effect. Conclusion CML/RAGE inhibits VSMC cholesterol efflux, induces vascular smooth muscle cell derived-foam cells formation.

Abstract:Aim Circulating visfatin levels has reflected the long-term survival in patients with ST-elevation myocardial infarction (STEMI). It postulated that higher visfatin would be linked to increased risk for atrial fibrillation (AF) and major adverse cardiovascular events (MACE) in patients with STEMI treated with primary percutaneous coronary intervention (PPCI). Methods 604 patients with acute STEMI underwent the PPCI were enrolled in the study. ELISA was used to measure plasma visfatin concentrations. One-year MACE and adverse events were compared between patients with and without new-onset AF after PPCI, and statistical analysis was used to respectively analyze the relationship between plasma visfatin level and MACE, plasma visfatin level and new-onset AF. Results From statistical data we learned that 42 of these patients had the new-onset AF (6.95%). The incidence rate of MACE in patients with new-onset AF was higher than patients without AF after PPCI within one year (45.24% vs 19.57%, P<0.05). Moreover, compared with patients with low plasma visfatin levels (≤14.5 μg/L), the patients with higher plasma visfatin levels (＞14.5 μg/L) had high rates of MACE (31.02% vs 9.56%, P<0.001) and new-onset AF (10.24% vs 2.94%, P<0.001). The multivariate Cox hazard regression model revealed that the plasma visfatin level was an independent predictor for occurrence of new-onset AF after PPCI one year (hazard ratio (HR) was 1.51, and 95% confidence interval (CI) was 1.03 to 2.23, P=0.021). Conclusion Plasma visfatin levels have a positive correlation with the incidence rate of new-onset AF and connection with MACE in patients with acute STEMI treated with PPCI.

Abstract:Cholesterol is crucial for the survival of cells, and cholesterol and its metabolites are involved in the pathogenesis of atherosclerosis, cancer, neurodegenerative diseases etc. Acyl-CoA:cholesterol acyltransferase (ACAT) catalyzes the esterification of cholesterol in cells, and plays an important role in the intracellular cholesterol homeostasis. ACAT is the therapeutic target of many diseases. This article will review the role of ACAT in cholesterol metabolism and associated diseases.

Abstract:Aim To investigate the effect of aerobic exercise on the microvascular rarefaction in spontaneously hypertensive rats (SHR) and explore whether the mechanism is related to the change of vascular endothelial growth factor (VEGF) expression. Methods Male SHR and Wistar-Kyoto rats (WKY), 12 weeks age, were randomly divided into a sedentary group (WKY-SED, SHR-SED), and an exercise group (WKY-EX, SHR-EX). WKY-SED and SHR-SED were used as normotensive control. Exercise groups were subjected to a 12-week treadmill training protocol:20 m/min (about 55%～65% of maximal aerobic velocity), 0% grade, 60 min/day, 5 d/w. Immunohistochemical staining was used to observe distribution and changing of VEGF in rat gastrocnemius muscle of each group. Western blot was applied to examine the expression level of VEGF in rat gastrocnemius muscle. HE staining was used to observe the skeletal mucle capillaries of each group. Results The positive immunoactivity of VEGF in SHR-SED group was significantly lower than WKY-SED group (P<0.01). After 12 weeks, exercise training markedly increased the VEGF expression in rat gastrocnemius muscle (P<0.01). No significant differences were observed in protein expression of VEGF between SHR-SED and WKY-SED (P＞0.05). After 12 weeks, the protein expression in SHR-EX was significantly higher than SHR-SED (P<0.05). Skeletal muscle capillaries were observed by light microscope, the result showed that the capillary/fiber of SHR-SED was significantly lower than WKY-SED (P<0.01). After 12 weeks' training, the capillary/fiber showed an obvious increasing in SHR-EX group. Conclusions Microvascular rarefaction is a possible reason that leads to hypertension. Aerobic exercise can effectively increase the expression of VEGF in the skeletal muscle and accelerate angiogenesis, which may be one of the mechanisms of exercise reversing hypertension.

Abstract:Aim To investigate whether the onion essential oil from Allium cepa has a protective effect on the oxidative damage cell model of atherosclerosis (As) and whether the protective mechanism is related to the regulation of janus kinase/signal transduction and transcriptional activator (JAK/STAT) pathway and NF-кB pathway. Methods Volatile oil extracting equipment was used to extract the essential oil from fresh Allium cepa. GC-MS was used to analyze the liposoluble components of the essential oil from Allium cepa. MTT method was used to detect the effect of the essential oil at different concentration and time on the cell proliferation of human umbilical vein endothelial cell (HUVEC). The HUVEC damage model has been established by using 80 mg/L oxidized low density lipoprotein (ox-LDL) to HUEVC for 24 h. The cell was stained by oil red O. The total cholesterol (TC) was detected by commercial kits. Free cholesterol (FC)/TC was more than or equal to 50%, which indicated whether HUVEC damage has been formed. ELISA kit, reactive oxygen species (ROS) kit and superoxide dismutase (SOD) kit were used to study the onion essential oil of anti-atherosclerosis effect and mechanism in vitro. Results 9.9008 g essential oil was extracted from 26615.5 g fresh Allium cepa, and the average yield of essential oil was 0.372‰. GC-MS analysis showed that the total relative content of organic sulfide in the essential oil was 47.64%, among which the content of 2-thiazolidinethione among the organic sulfide compounds was the highest (12.52%), followed by 1,3,5-trithiane (4.42%) and diallyldisulphide (3.82%). The cell proliferation was the best after 200 mg/L onion essential oil had acted on HUEVC for 6 h. HUEVC was induced by 80 mg/L ox-LDL for 24 h, the cells had a large number of red dye particles and FC/TC was not less than 50% of HUVEC cells which showed lipid accumulation and cholesterol and triglyceride metabolism has caused HUVEC damage. The onion essential oil significantly reduced HUVEC damage model secretion of inflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) (P<0.01) and the produce of ROS, promoted the produce of anti-inflammatory factor IL-10 (P<0.01) and increased activity of intracellular SOD (P<0.05). Conclusion The onion essential oil produced in this study has protective effect on the HUVEC damage model induced by ox-LDL.

Abstract:Aim To evaluate the impact of atherogenic index of plasma (AIP) on prognosis in patients with premature acute coronary syndrome (ACS) after percutaneous coronary intervention (PCI). Methods A total of 208 premature ACS patients (male≤55 years old, female≤65 years old ) with PCI were studied. According to the cutpoint of AIP at 0.06, the patients were classified as atherogenic phenom group (n=155) and non-atherogenic phenom group (n=53). The patients were followed-up for 1 year. The basic clinical characteristics, incidences of mortality and major adverse cardiovascular and cerebravascular events (MACCE) were compared between two groups. Results Atherogenic phenom group had higher mortality, higher rate of MACCE than those in non-atherogenic phenom group (11.0% vs 1.9%, P＜0.05; 24.5% vs 11.3%, P＜0.05, respectively). Multiple factor Logistic analysis revealed that AIP was independently associated with MACCE among patients with premature ACS at 1 year after PCI(OR=2.9,5%CI:1.426~3.209, P<0.05). Multiple Cox proportional hazard analysis showed that AIP was an independent predictor for all cause death among patients with premature ACS at 1 year after PCI (HR=1.7,5%CI:1.137~2.879, P<0.05). Conclusion AIP has a certain predictive value for all-cause death and MACCE after PCI in patients with premature ACS.

Abstract:Aim To investigate the relationship between uric acid, blood lipids and ambulatory arterial stiffness index (AASI). Methods A total of 280 subjects without taking anti-hypertensive medication from Fuwai Hospital during January to June, 2011 were enrolled. Ambulatory blood pressure monitoring (ABPM) and blood biochemical examination were performed among all subjects. AASI was calculated as 1 minus the regression coefficient of diastolic blood pressure on systolic blood pressure according to ABPM recordings. Pearson correlation analysis and stepwise multivariable linear regression models were used to assess the relationships between AASI and biomarkers. Results Among 280 subjects, 161 were men, 138 were hypertensive, and the average age was (50.4±13.3) years old. Pearson correlation analysis indicated that AASI was related to serum uric acid (r=0.168, P=0.049), total cholesterol (r=0.269, P=0.001), low-density lipoprotein cholesterol (LDLC) (r=0.223, P=0.009) in hypertensive participants. Stepwise multivariable linear regression analysis indicated that AASI was independently associated with serum uric acid (β=0.219, P=0.007) and total cholesterol (β=0.226, P=0.005) in hypertensive participants. Conclusion Serum uric acid and total cholesterol are independently correlated to AASI in hypertensive patients.

Abstract:Aim To investigate the correlation between flow mediated dilatation (FMD) and the degree of coronary artery lesion and provide a clinical basis for the detection of vascular endothelial function in early coronary heart disease. Methods A total of 130 patients with suspected coronary heart disease (CHD) who underwent coronary angiography (CAG) were recruited. The FMD was performed before CAG. The patients were divided into CHD group and non-CHD group according to the result of CAG. Meanwhile, the differences were analyzed about FMD in different lesion degree of coronary artery. ROC curves were drawn to find the optimal cut-off value for diagnosis of CHD. Results Systolic blood pressure, homocysteine and HbA1c in CHD group were higher than those in non-CHD group (P＜0.01 or P＜0.05). The FMD in CHD group was significantly lower than that in non-CHD group (P＜0.01). Further subgroup analysis showed that the FMD in normal group was higher than that in coronary atherosclerosis group, single branch lesion group, double branch lesion group and three branch lesion group (P＜0.05 or P＜0.01), the FMD in coronary atherosclerosis group was respectively higher than that in double branch lesion group and three branch lesion group (all P＜0.01), the FMD in single branch lesion group was higher than that in three branch lesion group (P＜0.05). Logistic regression analysis indicated that the FMD was an independent predictor of CHD (OR＝0.2,5%CI was 0.068～0.487, P＜0.01). ROC curve analysis showed that FMD≤6.05% was the best cut-off value for predicting CHD, and the sensitivity was 91.1%, the specificity was 80.4%, positive predictive value was 87.8% and the negative predictive value was 85.4%. Conclusion The FMD can better reflect the degree of coronary artery lesions and has a good predictive value for the diagnosis of coronary heart disease.

Abstract:Aim To elucidate the characteristics of carotid plaques in patients between acute coronary syndrome (ACS) and atherothrombotic cerebral infarction (ACI). Methods 100 patients (50 patients with single-vessel coronary artery disease (SVD) and the others with multivessel coronary artery disease (MVD) ) with ACS and fifty-six patients with ACI were enrolled in this study, carotid artery ultrasound-based atherosclerotic parameters of all patients were measured. Results Carotid atherosclerotic plaques in patients with ACS and ACI were multiple in the bilateral carotid sinus, and more common with hard spot. The incidence of plaque in ACS patients with MVD was higher than that SVD (P<0.05), similar to those with ACI. The incidence of unstable plaque and stenosis in patients with MVD and ACI was higher than those with SVD (P<0.05). Conclusions High-frequency ultrasound findings showed similar characteristics of carotid plaque in ACS and ACI patients. The similarity of cervical plaque features in MVD patients was more obvious.

Abstract:Aim To investigate the association between the expression of Beclin-1 and ischemic cardiomyopathy.Methods Participants in case group (n＝80) were selected from autopsy cases accepted by Forensic Judicial Expertise Center of Guizhou Medical University from July 2013 to December 2016, pathologically confirmed to have suffered “ischemic cardiomyopathy”. And participants in control group (n＝122), the ones without myocardial tissue lesions, were selected from autopsy cases accepted during the same period. Beclin-1 was detected in all samples by immunohistochemistry.And the effect of Beclin-1 expression on ischemic cardiomyopathy was evaluated by establishing multiple Generalized Additive Models (GAM), adjusting the confounding factors such as gender, age, height, body mass index (BMI), left ventricular wall thickness (LVWT), right ventricular wall thickness (RVWT), alcohol abuse, Gensini score, previous myocardial infarction history (MI), hypertension, chronic kidney disease (CKD), and chronic respiratory disease. Results This research demonstrated that the expression of Beclin-1 in cardiomyocytes was positively correlated with ischemic cardiomyopathy (OR＝1.2,5％CI was 1.1～1.3, P＜0.001). After adjusting for other confounders, Beclin-1's effect on ischemic cardiomyopathy was that the expression of Beclin-1 increased by 0.01 units, and the risk of ischemic cardiomyopathy increased by 20%. Conclusion Beclin-1 is an independent risk factor for ischemic cardiomyopathy.

Abstract:Aim To establish a simplified method for extraction of lipids from high density lipoprotein (HDL), qualify and quantify lipids by liquid chromatography tandem mass spectrometry (LC-MS/MS). Methods The acidic methanol precipitation combined with ultrasonic extraction method was used to extract lipids, and the supernatant after centrifugation was analyzed directly by LC-MS/MS with multiple reaction monitoring mode. Results The acidic extraction method was effective and time saving, and the established LC-MS/MS method using lipids standards could simultaneously quantify more than 30 kinds of lipids, including sphingolipid, sphigomyelin, ceramide and oxidized phosphatidylcholine.Conclusions Acidic methanol precipitation combined with ultrasonic extraction can effectively isolate lipids from HDL, and the extract after centrifugation can be determined by LC-MS/MS directly. This method is effective with good reproducibility, and is suitable for quick quantitative analysis of a great deal of biological liquid samples.

Abstract:Vascular calcification is associated with an increased risk of death from cardiovascular disease, however vascular calcification is common in patients with coronary artery disease. The pathogenesis of vascular calcification is similar to the process development of bone and chondrogenesis. Vascular smooth muscle cells transdifferentiate into osteoblast-like cells, the imbalance of calcium and phosphorus adjustment, osteoclast activity and the decrease of mineral absorption capacity play an important role in promoting vascular calcification. It is now found that microRNA are involved in the process of vascular calcification by directing vascular smooth muscle cells for complex genetic reprogramming and other cellular functional responses associated with vascular calcification. This article presents in detail the important regulation that has so far involved microRNA-mediated vascular calcification.

Abstract:Because of the usually asymptomatic nature of patients with coronary myocardial bridge, this anatomical abnormality has long been considered a benign variant. However, with the popularity of coronary angiography and clinical research, more and more scholars believe that myocardial bridge may be serious complications, and even threaten human life. At present, its pathogenesis, diagnosis and treatment have received extensive attention and research, but there is no clear guide to guide the clinical diagnosis and treatment. This review summarizes the incidence, pathophysiology, diagnosis and treatment of myocardial bridge and the research progress, and provides the basis for the standardized diagnosis and treatment of myocardial bridge to benefit more and more patients.

Abstract:CD36 is a multifunctional transmembrane glycoprotein that binds to oxidized low density lipoprotein to induce monocyte transformation into foam cells and promotes the formation of atherosclerosis through inflammatory response, oxidative stress, platelet activation, macrophage trapping. Inhibition of CD36 expression or interference with its associated signaling pathways can significantly alleviate the severity of atherosclerosis. In addition, the high expression of CD36 in the tongue, nasal cavity, small intestine and brain promotes the body's intake and absorption of lipids, and increases the risk factors of metabolic diseases. Serum soluble CD36 is found as a component of circulating microparticles and may be as a predictor for atherosclerotic disease.