Abstract:Cardiovascular disease (CVD) is one of the most important complication and the primary cause of death in chronic kidney disease (CKD) patients. Early prevention and treatment of CVD in CKD patients, depend on the evaluation of CVD presentations and risk factors for CVD, derived from end stage renal disease.

Abstract:Vascular calcification is a process of arterial wall mesenchymal cells, especially smooth muscle cells, transforming into the phenotype of chondroblast and osteoblast under the effect of various pathological factors. It mediates the process of abnormal calcium deposition in the vascular wall. Vascular calcification includes various pathological types such as intimal calcification, medial calcification, and valve calcification. With the increase of aging population, especially the prevalence rate of diabetes, atherosclerosis as well as chronic kidney disease continues to rise, the vascular calcification derived from which is gradually becoming a key disease spectrum affecting the health of our people. For that reason, the thesis starts from the origin, evolution and prognosis of vascular calcification, systematically discusses some controversial and hotspot issues in the process of prognosis such as bone and blood vessel, active and passive, intimal and medial calcification, microcalcification and macrocalcification, autophagy, endoplasmic reticulum stress and non-coding RNA.It hopes to promote the advancement of basic and clinical research on vascular calcification through the joint efforts of experts who devote in the calcification.

Abstract:Aim To explore whether 1,5-(OH)₂D₃ can regulate the autophagy and calcification of human umbilical vein endothelial cells (HUVEC) and the calcification of mouse atherosclerotic plaque through Ca²⁺/CaM signal. Methods (1) HUVEC were incubated in high fat microenvironment to stimulate autophagy. The autophagy level was detected by Western blot, fluorescence microscopy, transmission electron microscopy after HUVEC being stimulated by 1,5-(OH)₂D₃. Lentivious were used to overexpress or silence calmodulin (CaM), and the calcification was tested by Western blot,calcium ion detection kit, Alizarin red. (2) ApoE－/－ mice fed with high fat diet were randomly divided into 3 groups:control group (1,5-(OH)₂D₃ 2.5 μg/(kg·d) dissolved in 0.1 mL soybean oil, Gavage)； CaM overexpressed group (control +CaM overexpressed lentivirus, Caudal vein injection) and CaM silenced group (control+CaM silenced lentivirus, Caudal vein injection). Scanning electron microscope was used to observe autophagosomes and calcification of the aorta. HE and Von Kossa staining were used to detect the As degree and calcification of the aorta, respectively. Results The autophagy was promted and the expression of Ca²⁺ and CaM were up-regulated after HUVEC being stimulated by 1,5-(OH)₂D₃. Compared with the control group, CaM overexpressed group showed enhanced autophagy and reduced calcification in HUVEC and As plaque in ApoE－/－mice. While the CaM silencing group showed autophagy dysfunction, increased calcium depositions in HUVEC and plaques. Conclusion 1,5-(OH)₂D₃ regulates HUVEC autophagy to inhibit cell calcification and plaque calcification through Ca²⁺/CaM signaling.

Abstract:Aim To determine the effect of interplay between endoplasmic reticulum stress (ERS) and autophagy on vascular calcification (VC). Methods VC model in rats was induced by vitamin D3 plus nicotine. The deposition of calcium in aorta was determined by alizarin red staining and assessment of calcium content. The protein levels in aorta were measured by Western blot. Results Compared with control group, deposition of calcium in aorta was increased in VC group. The protein levels of SM-22α and calponin in calcified vessels were decreased, whereas that of bone morphogenetic protein 2 (BMP-2) and RUNX2 was increased. ERS and autophagy was activated in calcified aorta, shown by increased levels of glucose regulated protein (GRP78), C/EBP homogous protein (CHOP), LC3Ⅱ and Beclin-1. Tunicamycin (10 μg/kg per day), an ERS inducer exacerbated VC and transdifferation of vascular smooth muscle cells from contracted-like to osteoblasts-like phenotype. It also promoted ERS and auphagy. 3-methyladenine (10 mg/kg per day), an autophagy inhibitor prevented stimulation of autophagy, but promoted ERS activation in calcified aorta, while it also exaggerated VC and transdifferation of vascular smooth muscle cells. Conclusion The interplay between ERS and autophagy plays an important role in progression of VC.

Abstract:Aim To analyze the characteristics of vascular calcification (VC) in aortic dissection (AD) of patients with essential hypertension. Methods From January 2010 to December 5,0 patients with essential hypertension diagnosed as AD by computed tomography angiography (CTA) in Sun Yat-sen Memorial Hospital of Sun Yat-sen University were selected as the AD group, and 30 cases of essential hypertension and aortic CTA showed no aortic disease were as the control group. Characteristics of aortic calcification in two groups of people and patients with different Stanford types of AD were analyzed by CTA and Agatston scoring system. Results Compared with the control group, the VC incidence of descending aorta increased and the VC incidence of aortic arch decreased in AD group (P＜0.05). There was no significant difference between the two groups in the Agatston score and the incidence of VC with different degrees (P＞0.05). Patients in group AD were divided into two subgroups:Stanford A type and Stanford B type. There was no significant differences in the clinical data and the incidences of VC in aorta and different parts of the two subgroups (P＞0.05).Multivariate Logistic regression analysis showed that the factors associated with AD lesions were:low density lipoprotein cholesterol (OR 3.7,5%CI 1.234-9.354, P＝0.018), irregular use of antihypertensive drugs (OR 20.7,5%CI 2.326-179.233, P＝0.006), VC incidence of aortic arch (OR 0.7,5%CI 0.002-0.357, P＝0.006) and VC incidence of descending aorta (OR 35.6,5%CI 2.475-512.27, P＝0.009). Conclusion The differences of calcification in different parts of aorta in patients with essential hypertension may be an important link of hypertension in promoting AD lesion.

Abstract:Vascular calcification is an active cell- and gene-regulated process, involving imbalance between promoters and inhibitors of vascular calcification. It commonly occurs in patients with advanced atherosclerosis and is highly related with diabetes, chronic kidney diseases and aging. Osteogenic differentiation of vascular cells is the key contributor to vascular calcification, but the cell sources of vascular calcification remain unclear. It has been thought that the origins of cells regulating vascular calcification may include vascular smooth muscle cell, endothelial cell, pericyte, macrophage and progenitor cell. This review focuses on the regulatory role of cells in vascular calcification.

Abstract:Calcific aortic valve disease is frequently observed in the elderly. Currently, there is no treatment to reverse established calcific aortic valve disease. The responsible underlying molecular mechanisms have yet to be fully elucidated. Previous studies have demonstrated that calcific aortic valve disease shares many similarities to that of bone formation and bone metabolisms. lncRNAs are defined as non-coding transcripts longer than 200 nucleotides, with important biological functions. Emerging data have shown that lncRNAs differentially express in calcified aortic valves compared to normal control, which contribute significantly to the pathogenesis of calcific aortic valve disease through a number of distinct mechanisms. This study reviews here the current progress of lncRNAs and calcific aortic valve disease in recent years. These studies shed new lights on pathogenic mechanisms of calcific aortic valve disease.

Abstract:Aim To investigate the effect of down-regulation of miR-92a on the secretion of inflammatory mediators and lipid accumulation induced by oxidized low density lipoprotein (ox-LDL) in RAW264.7 cells and its mechanism.Methods RAW264.7 cells were treated with ox-LDL of different concentrations (0,5, 0,0 mg/L) for 24 hours or with 100 mg/L ox-LDL for 0,2 and 24 hours respectively. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the effect of ox-LDL on the expression of miR-92a. A RAW264.7 cell line with low expression of miR-92a was established and treated with 100 mg/L ox-LDL. Effects of down-regulation of miR-92a expression on expressions of inflammatory factors inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) and IL-1β, lipid accumulation and expressions of signal transducer and activator of transcription-3 (STAT3) signal pathway-related proteins p-STAT3 and STAT3 in RAW264.7 cells induced by ox-LDL, were detected by qRT-PCR, oil red O staining, nitric oxide (NO) fluorescence probe (DAF-FMDA) and Western blot. A RAW264.7 cell line with low expressions of protein inhibitor of activated STAT3 (PIAS3) and miR-92a was established and treated with 100 mg/L ox-LDL. The effects of silencing PIAS3 expression on inflammatory factors, lipid accumulation and STAT3 signaling pathway were observed in RAW264.7 cells with low expression of miR-92a induced by ox-LDL. TargetScan software prediction and dual luciferase reporting experiment were used to verify the target relationship between miR-92a and PIAS3. Results With the increase of the action time and concentration of ox-LDL, the expression of miR-92a was increased in RAW264.7 cells. The dual luciferase reporting experiment confirmed that PIAS3 was the target gene of miR-92a. After ox-LDL treatment, the expressions of miR-92a, iNOS, IL-6, IL-1β and p-STAT3 were increased, NO release and intracellular lipid droplets were increased in RAW264.7 cells, while the expression of PIAS3 was decreased. This regulatory effect could be inhibited by anti-microRNA-92a, and the inhibition of anti-microRNA-92a could be restored after PIAS3 silence. Conclusion Down-regulation of miR-92a can inhibit ox-LDL induced secretion of inflammatory mediators and lipid accumulation in RAW264.7 cells by regulating PIAS3.

Abstract:Aim To investigate the regulatory role of miR-146a in the senescence of human umbilical vein endothelial cells and its mechanism. Methods The replication aging model of human umbilical vein endothelial ECV-304 cells was established. Senescence-associated β-galactosidase (SA-β-gal) method was used to detect the ratio of senescent cells in different generations cells (P2, P6, P9, P12, P15 generation); real time quantitative PCR method was used to detect the level of miR-146a in different generations cells (P2, P6, P9, P12, P15 generation); Target gene prediction software was used to predict the target gene of miR-146a. The cells in P12 generation were transfected with Pre-miR146a, Pre-miR146a control, Anti-miR146a , and Anti-miR146a control; SA-β-gal method was used to detect the ratio of senescent cells; Western blot was used to detect the protein expression of NADPH oxidase 4(NOX4). Results The ratio of senescent cells in P6, P9, P12 and P15 generations was significantly higher than cells in P2 generation (P<0.05). And the expression of miR-146a in P6, P9, P12 and P15 generations was significantly lower than cells in P2 generation (P<0.05). Predicting the target gene of miR-146a was NOX4. After transfection, the ratio of senescent cells in Pre-miR146a group was significantly lower than control group (P<0.05), but it was significantly higher in Anti-miR146a group (P<0.05). The expression of NOX4 protein in Pre-miR146a group was significantly lower than control group (P<0.05), but it was significantly higher in Anti-miR146a group than control group (P<0.05). Conclusion The expression of miR-146a decreased in the aging human umbilical vein endothelial ECV-304 cells, the up-regulation of miR-146a expression can inhibit ECV-304 senescence, and its mechanism is related to the regulation of NOX expression.

Abstract:Aim To investigate the effect of asiatic acid on acute myocardial infarction (AMI) rats and its mechanism. Methods Healthy SD rats were used to establish acute myocardial infarction model by ligation of anterior descending branch of left coronary artery, then the successful model rats were randomly divided into model group, low, middle and high dose of asiatic acid group, 10 rats in each group. Sham group only threaded without ligation. Asiatic acid group were given asiatic acid 25 mg/kg, 50 mg/kg and 100 mg/kg by gavage, once a day for 28 consecutive days. The changes of ST segment of electrocardiogram were recorded before operation, immediately after operation, 1,3, 7,0, 4,1 and 28 days after administration. Myocardial apoptosis was detected by TUNEL, morphological and pathological changes of myocardial tissue were observed by HE staining, serum lactate dehydrogenase (LDH), creatine kinase isoenzyme (CK-MB) and superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by ELISA. The expression of Nrf2, HO-1 and NF-κB mRNA were determined by RT-PCR, the expression of Nrf2, HO-1 and NF-κB protein was determined by Western blot. Results The electrocardiogram ST segment of acute myocardial infarction rats was significantly elevated after model establishment, the electrocardiogram ST segment of myocardial infarction rats after asiatica acid treatment showed a downward trend in varying degrees (P＜0.05). HE staining showed that myocardial cells in the model group were disordered and infiltrated with a large number of inflammatory cells, and the nuclei dissolved, disappeared or even necrosed. The degree of myocardial cell disorder, the phenomena of cell nucleolysis and the infiltration of inflammatory cells were alleviated in all dose groups of asiatica acid. Compared with sham group, apoptotic myocardial cells increased significantly (P＜0.05), serum levels of CK-MB, LDH, MDA decreased significantly and SOD increased significantly (P＜0.05), the expression of Nrf2, HO-1 mRNA and protein decreased significantly (P＜0.05), the expression of NF-κB mRNA and protein increased significantly in the model group (P＜0.05). Compared with model group, apoptotic myocardial cells decreased significantly (P＜0.05), serum levels of CK-MB, LDH, MDA increased significantly and SOD decreased significantly (P＜0.05), the expression of Nrf2, HO-1 mRNA and protein increased significantly (P＜0.05), the expression of NF-κB mRNA and protein decreased significantly in all dose groups of asiatica acid (P＜0.05). Conclusion Asiatic acid can improve myocardial injury in rats with acute myocardial infarction, which may be achieved by regulating Nrf2/HO-1 and NF-κB signaling pathways.

Abstract:Aim To evaluate the characteristics of cardiac structure and function in atherosclerotic adults during quietness and exercise in order to assess the risk and adverse consequences of cardiac changes during exercise. Methods 107 Beijing residents aged 40-59, including 21 males and 86 females, were selected. According to the criteria of brachial-ankle pulse wave velocity (baPWV), the participants were divided into normal arterial stiffness group (n＝71) and arteriosclerosis group (n＝36). After resting echocardiography, all participants performed a horizontal exercise test that the initial load was 25 W, increasing 25 W every two minutes. Echocardiography and blood pressure data were obtained at 30 seconds before the end of each exercise and convalescence. Results (1)Compared with the normal arterial stiffness group, the arteriosclerosis group showed increasing interventricular septum thickness at end-diastole, left ventricular posterior wall thickness at end-diastole (LVPWd), left ventricular mass, left ventricular mass index, ejection fraction (EF), fractional shortening (FS) (P＜0.05) and decreasing peak flow velocity of early diastole/peak flow velocity of atrial contraction (E/A) (P＜0.05). (2)Left ventricular hypertrophy in the arteriosclerosis group was much higher than the normal arterial stiffness group (P＜0.05). (3)BaPWV was positively correlated with heart rate, EF, stroke volume and FS (P＜0.05). (4)During exercise, left ventricular end-diastolic diameter, LVPWd and systolic blood pressure in atherosclerosis group were significantly higher than those in normal arterial stiffness group (P＜0.05). Conclusions Cardiac manifestations of atherosclerosis individuals include thickening of ventricular wall, increase of left ventricular mass, enhancement of systolic function, decrease of diastolic function, and remodeling of left ventricular geometry, and the change of systolic function is independent on the change of blood pressure. Compared with heart changes, arteriosclerotic people should prevent excessive blood pressure during exercise.

Abstract:Aim To investigate the possible relationship between variant CD33 rs3865444 and serum lipid levels in long-lived population inhabiting along Hongshuihe River Basin in Guangxi province. Methods Genotyping of CD33 rs3865444 locus was performed by improved multi-temperature ligase detection reaction technique (iMLDR) for long-lived individuals (long-lived group, aged≥90 years, n＝645), non-long-lived elderly (local control group, aged 60～77 years, n＝674) residing in Hongshuihe River Basin and non-long-lived elderly living in Hezhou (non-local control group, aged 60～77 years, n＝662), a town about 600 km from Guangxi Honeshuihe. Association of CD33 rs3865444 genotypes with lipids was analyzed thereafter. Results The frequencies of A allele and its genotype (CA/AA) of females in Hongshuihe River Basin, both long-lived group and long control group, were significantly higher than those of non-local controls; CA/AA genotype of females was higher than that of males in the long-lived group, while this status was opposite between gender in the non-local controls. The levels of TC, LDLC and ApoB of A allele carriers in males in long-lived group were dramatically greater than those of non-A allele carriers (P＜0.0001), while HDLC level of A carriers was lower than non-A carriers in females of local controls; by contrary, HDLC and ApoA levels of females were higher than those of non-A carriers in the non-local controls. Conclusions CD33 rs3865444 polymorphism correlates with lipid levels to some extent, which is influenced by sex and other environmental factors. The enrichment of A allele of CD33 rs3865444 locus in the long-lived females needs further clarification.

Abstract:Aim To evaluate the predictive value of low density lipoprotein subfractions detected by lipoprint method for carotid artery intima-media thickness (CA-IMT). Methods A retrospective analysis of 175 patients in the department of cardiology of our hospital, the average age was 67 years, including 106 males (60.6%) and 69 females (39.4%). In all patients lipoprint lipoprotein classification detection system was used for low density lipoprotein(LDL) subfractions of cholesterol and four types of blood lipids, the color Doppler ultrasound detector was used to measure the thickness of the carotid artery intima-media, the differences in blood lipid levels and CA-IMT between groups under traditional risk factors were compared. Linear correlation analysis was used to evaluate the relationship between CA-IMT and traditional risk factors and low-density lipoprotein subfractions. Logistic regression analysis was used to evaluate the predictive value of low-density lipoprotein subfractions in CA-IMT. Results Compared with males, the LDL4 level was lower in female patients (P<0.05), and the CA-IMT was lower than that in males (P<0.01). The small, dense low-density lipoprotein(sdLDL), subfractions and CA-IMT of patients with traditional risk factors such as hypertension and diabetes were higher than the normal group, but the difference was not statistically significant (P>0.05). Correlation analysis showed that CA-IMT was negatively correlated with gender (P<0.05), and positively correlated with total cholesterol (TC), LDL and non high density lipoprotein (non-HDL) (P<0.05)； and it was positively correlatied with sdLDL, low-density lipoprotein subfractions LDL3, LDL4 (P<0.01). CA-IMT was also positively correlated with traditional risk factors smoking (P<0.05). Logistic regression analysis showed that LDL, sdLDL and sdLDL subfractions LDL3 and LDL4 were independent predictors of CA-IMT thickening. Conclusion Small, dense low-density lipoprotein and its subfractions have a correlation with CA-IMT and have predictive value for atherosclerosis.

Abstract:Aim To investigate the relationship among monocyte/high density lipoprotein cholesterol ratio and inflammatory adipocytokines with the severity of coronary artery stenosis. And then the diagnostic value of the new markers on coronary artery disease (CAD) was analyzed. Methods A total of 168 inpatients with CAD and a control group of 74 cases without significant coronary artery stenosis were consecutively enrolled from January 2017 to August 2017 in the division of cardiology in our hospital. All the demographic and clinical data of the subjects were collected by the physicians and master degree candidates in the division of cardiology. The concentrations of plasma inflammatory adipocytokines were measured by ELISA. Results Monocyte/high density lipoprotein cholesterol ratio, plasma levels of PTX3, RBP4, GAL-3 and PAI-1 were higher, while the plasma levels of IL-37, NTN1 and ADP were lower in CAD group than those in the control group. Monocyte/high density lipoprotein cholesterol ratio was significantly correlated with the coronary Gensini score and inflammatory adipocytokines. In addition, the area under the receiver operating characteristic curve (ROC-AUC) of monocyte/high density lipoprotein cholesterol ratio was 0.676. Conclusion Monocyte/high density lipoprotein cholesterol ratio and the adipocytokines may be enlightening diagnostic markers of CAD, which also will be helpful in assessing the severity of coronary artery stenosis in clinical practice.

Abstract:Diabetes is a worldwide epidemic disease and closely related to cardiovascular diseases such as atherosclerosis (As). In a long-term hyperglycemic environment, a series of complex non-enzymatic glycosylation reactions of proteins, fatty acids and other macromolecular substances in diabetic patients result in the formation of advanced glycosylation end products (AGE). Glycosylated albumin (GA) is the most important form of AGE, which promotes the progression of atherosclerosis by binding to glycosylated product receptor (RAGE) to induce oxidative stress, inflammation and endoplasmic reticulum stress. Therefore, in-depth study of the mechanism of GA-induced atherosclerosis is of great significance to reduce the incidence of cardiovascular events in diabetic patients and delay the disease process. At present, the existed researches have partially revealed its specific mechanisms, and this article reviews the roles of GA in the occurrence and development of atherosclerosis.

Abstract:Coronary heart disease is one of the leading diseases that threaten the health of urban and rural residents in China. Recent studies have shown that chronic inflammation is the core pathogenic mechanism of the induction and development of atherosclerosis, and various pathways of inflammatory response have become targets for the treatment of coronary heart disease. However, no specific target anti-inflammatory drugs have been recommended for secondary prevention of coronary heart disease. The CANOTOS study, published at the European Society of Cardiology Congress 2017 (ESC 2017), confirmed that canakinumab significantly reduces cardiovascular risk in the target population through anti-inflammatory action. This landmark study reveals that anti-inflammatory therapy is a new chapter in the treatment of coronary heart disease after the age of traditional lipid-lowering and antithrombotic therapy. In recent years, rapid progress has been made in the research of targeted anti-inflammatory drugs for various types of coronary heart disease. The current progress in the research of targeted anti-inflammatory therapy for coronary heart disease is reviewed.

Abstract:Scavenger receptor type B class I (SR-BI) is the first glycoprotein defined as a high density lipoprotein (HDL) receptor on the cell membrane, which mediates the selective uptake of high density lipoprotein cholesterol (HDL) by cells, affecting cell cholesterol balance and inflammatory status. Liver SR-BI plays an important role in reverse cholesterol transport, and is closely related to HDL metabolism and its role in anti-atherogenesis. It has been found that humans have SR-BI gene polymorphisms. This review focuses on SR-BI structure, function, regulation and its role on atherosclerosis.

Abstract:Familial hypercholesterolemia (FH) is a common autosomal dominant metabolic disease with an incidence of heterozygous FH:1/(137～350) and homozygous FH:1/(160000～300000). Early diagnosis and treatment are critical to the FH population. But the detection rate of FH is even less than 1% in more than 190 countries. Now the commonly used diagnostic criteria for FH mainly rely on cholesterol level test and patient health survey information. The rate of diagnosis/detection without genetic tests is very limited, so genetic testing is considered to be the gold standard for FH diagnosis. Positive results can be confirmed directly. Common international strategies for FH gene screening include uniform screening(UniS), cascade screening (CasS), reverse cascade screening(RCS), targeted screening (TarS) and e-health record identification(EHRI). At present, there is no FH epidemiological investigation at the national level in China. There is no official national conditions and population screening program and diagnosis or treatment guidelines issued, making our country lag behind the international standard in this respect. It is necessary to obtain the appropriate gene testing method and screening program as soon as possible to improve the detection rate of FH in our country, and to ensure that the patients can get the appropriate treatment in combination with clinical practice, so as to benefit many patients with FH to the greatest extent.