Abstract:The highly dynamic tight junctions between the endoplasmic reticulum and mitochondria are known as mitochondria-associated ER membranes. These domains are essential for basic biological processes, including lipid metabolism, calcium homeostasis, mitochondrial dynamics, autophagy and apoptosis, endoplasmic reticulum stress and inflammation. Many studies have proved the abnormal amount, structure or function of mitochondria-associated ER membranes are related to the occurrence and development of cardiovascular diseases. This paper summarized the functions of mitochondria-associated ER membranes and its roles and possible mechanism in cardiovascular diseases, and provided theoretical references for mitochondria-associated ER membranes to become new targets for cardiovascular therapy.

Abstract:Aim To investigate the role and mechanism of Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil forming protein kinase (ROCK) pathway in hypoxia/reoxygenation (H/R)-induced injury in human cardiac AC16 cells. Methods AC16 cells cultured in vitro were divided into control group (normal culture), H/R group (construction of H/R model), H/R＋NC-siRNA group (H/R model was constructed after transfection of NC-siRNA) and H/R＋RhoA-siRNA group (H/R model was established after transfection of RhoA-siRNA), MTT assay was used to detect the survival rate of AC16 cells, the apoptosis rate of AC16 cells was detected by flow cytometry, the activities of lactate dehydrogenase (LDH) and Caspase-3 in AC16 cells were detected by colorimetry, the levels of interleukin-6 (IL-6), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the supernatant of AC16 cells were detected by ELISA, the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in AC16 cells were detected by kit, the mRNA expression levels of RhoA, ROCK1, ROCK2, nuclear factor-κB (NF-κB) p65 and IKB kinase (IKK) in AC16 cells were detected by real-time fluorescence quantitative PCR, the protein expression levels of RhoA, ROCK1, ROCK2, NF-κB p65, phosphorylated NF-κB (p-NF-κB) p65 and IKK in AC16 cells were detected by Western blot. Results Compared with those in the control group, the cell survival rate and SOD level in H/R group were significantly lower, apoptosis rate, LDH activity, Caspase-3 activity, MDA level, IL-6, IL-1β and TNF-α levels in cell supernatant, RhoA, ROCK1, ROCK2, NF-κB p65, p-NF-κB p65, IKK mRNA and protein expression levels were significantly higher (P＜0.05). There was no significant difference in the above indexes between H/R＋NC-siRNA group and H/R group (P＞0.05). Compared with those in H/R＋NC-siRNA group, the cell survival rate and SOD level in H/R＋RhoA-siRNA group were significantly higher, apoptosis rate, LDH activity, Caspase-3 activity, MDA level, IL-6, IL-1β and TNF-α levels in cell supernatant, RhoA, ROCK1, ROCK2, NF-κB p65, p-NF-κB p65, IKK mRNA and protein expression levels were significantly lower (P＜0.05). Conclusion Targeted inhibition of RhoA/ROCK pathway can reduce H/R-induced cardiomyocyte injury by reducing inflammatory response, oxidative stress and apoptosis.

Abstract:Aim To investigate the protective effect and mechanism of procyanidin B₂ (PCB₂) on lipopolysaccharide (LPS)-induced cardiomyocyte injury. Methods Cardiomyocytes H9c2 were cultured normally, and induced by LPS to establish cell damage models. The model cells were treated with PCB₂ of 6.5,2.5 and 25.0 μmol/L, 25.0 μmol/L PCB₂ treatment model was then treated with NF-κB signaling pathway inhibitor PDTC treatment. Tetrazolium salt colorimetry (MTT) was used to detect cell survival; flow cytometry was used to detect cell apoptosis; enzyme-linked immunosorbent assay (ELISA) was used to detect cell tumor necrosis factor alpha (TNF-α), interleukin 1β (IL-1β) and interleukin 6 (IL-6) levels; malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) kits were used to detect MDA content and activity of SOD and GSH-Px, respectively; Western blot was used to detect the expression of nuclear factor-κB (NF-κB) and IκB-α in cells. Results Compared with control group, the cell survival rate of the LPS group was significantly reduced (P＜0.05); compared with LPS group, PCB₂ significantly increased the cell survival rate (P＜0.05). Compared with control group, the apoptosis rate was significantly increased in LPS group (P＜0.05); compared with LPS group, PCB₂ significantly reduced the apoptosis rate (P＜0.05). Compared with control group, the levels of TNF-α, IL-1β, and were significantly increased IL-6 in LPS group (P＜0.05); compared with LPS group, PCB₂ significantly reduced the levels of TNF-α, IL-1β, and IL-6 in the cells (P＜0.05). Compared with control group, the cell MDA content in LPS group was significantly increased, and the SOD and GSH-Px activities were significantly reduced (P＜0.05); compared with LPS group, PCB₂ significantly reduced the cellular MDA content and significantly increased the SOD and GSH-Px activities (P＜0.05). Compared with control group, the cell NF-κB protein expression in LPS group was significantly increased, and the IκB-α protein expression was significantly decreased (P＜0.05); compared with LPS group, PCB₂ significantly reduced the cell NF-κB protein expression and significantly increased IκB- α protein expression (P＜0.05). Compared with LPS+PCB₂ group, LPS+PCB₂₊PDTC significantly reduced the apoptosis rate, TNF-α, IL-1β, IL-6 and MDA content, and significantly increased the SOD and GSH-Px activities. Conclusion PCB₂ reduces the apoptosis rate, inflammation level and oxidative stress of cardiomyocytes induced by LPS, and improves cell survival rate, which may be related to inhibiting the activation of NF-κB signaling pathway.

Abstract:Aim To verify the effects and mechanism of the deubiquitin enzyme USP22 participating in the estrogen receptor α(ERα) mediated gene transcription on vascular calcification (VC). Methods Human aortic smooth muscle cells (hASMC) were vaccinated respectively in the normal DMEM, 1.25 mmol/L CAL DMEM, 2.5 mmol/L CAL DMEM, 5 mmol/L CAL DMEM culture for 0,3, 7,9 days, with a 1%/2% Alizarin red staining, and the inverted phase contrast microscope imaging; Under the condition of 5 mmol/L CAL cultivating 0,3, 7,9 days, Western blot experiment was used to test the expression of calcification associated protein bone morphogenetic protein-2 (BMP-2), receptor activator of nuclear factor κB ligand (RANKL), cyclooxygenase-2 (COX-2), growth arrest-specific protein 6 (Gas6), ERα; Luciferase double gene report experiment was used to detect USP22 whether to participate in the ERα mediated gene transcription in hASMC; With overexpression of USP22, Western blot test was used to detect the expression of calcification associated protein BMP-2, RANKL, COX-2, Gas6, ERα. Results hASMC had significant calcium deposition at the 9th day of 1.25 mmol/L CAL condition culturing. In the condition of 2.5 mmol/L CAL and 5 mmol/L CAL culture condition, hASMC had small amount of calcium deposition at the 7th day, but at the 9th day, hASMC had obvious calcium deposition; In 5 mmol/L CAL conditions to developing, the expression of the RANKL, BMP-2, USP22 protein increased; Luciferase double gene report experimental results demonstrated that USP22 inhibited ERα mediated target gene transcription regulation; With overexpression of USP22, Western blot test demonstrated decreased expression of Gas6 protein. Conclusion With the condition of 5 mmol/L CAL concentration culture time, the model of hASMC calcification is successfully established;USP22 may inhibit ERα mediated target genes Gas6 gene transcription, and then facilitates hASMC calcification.

Abstract:Aim To investigate the relationship between single nucleotide polymorphism (SNP) of ATG16L1 gene promoter sequence and acute myocardial infarction (AMI). Methods The ATG16L1 gene promoter was amplified and sequenced by polymerase chain reaction in 285 AMI patients and 296 controls using case-control method. Combined with DNA sequencing sequence and alignment SNPs database, data statistics and analysis were carried out. After using the Hardy-Weinberg balance test, the χ² test and t test were used for correlation analysis. Logistic regression was used to analyze the association of multiple risk factors and three SNPs loci with susceptibility to AMI. Haploview 4.2 software and SHEsis online software were used for linkage disequilibrium and haplotype analysis. TRANSFAC database was used to predict the binding sites of transcription factors that may be affected by SNPs. Results Multivariate Logistic regression analysis showed that male, smoking history, and hypertension were independent risk factors for AMI (P＜0.05), while high density lipoprotein cholesterol was a protective factor for AMI (P＜0.05). Among the three SNPs (rs1816753, rs12476635, rs2289477) in the promoter sequence of ATG16L1 gene, the TC genotype of rs1816753 was associated with AMI, which significantly increased the risk of AMI (OR=2.9,5%CI:1.130～5.615, P＝0.024). Haploview 4.2 software analysis showed that the three SNPs were strongly linked. Conclusion The SNPs of ATG16L1 gene promoter may be associated with the susceptibility to AMI, and the TC genotype of rs1816753 may be a genetic risk factor for AMI.

Abstract:Aim To explore the correlation between remnant cholesterol and severity of coronary artery stenosis in patients with prehypertension based on Gensini score. Methods The clinical data of 36 216 coronary angiography patients hospitalized in the Department of Cardiovascular Medicine of Northern Theater Command General Hospital from 2004 to 2014 were retrospectively analyzed, including 421 patients with prehypertension. There were 85 patients with Gensini score ≥20, and 85 patients with Gensini score <20 in this cohort were selected as the control group. The clinical characteristics of the two groups were observed and the relationship between remnant cholesterol and common blood lipid indexes and the degree of coronary artery stenosis in prehypertensive patients was investigated. The seventh report of the National Joint Committee on the prevention, detection, evaluation, and treatment of hypertension in the United States defines prehypertension as systolic blood pressure of 120 to 139 mmHg(1 mmHg＝0.133 kPa) and/or diastolic blood pressure of 80 to 89 mmHg. The European Atherosclerosis Society (EAS) defines remnant cholesterol as total cholesterol－(high density lipoprotein cholesterol＋low density lipoprotein cholesterol). Remnant cholesterol is the amount of cholesterol in triglyceride-rich lipoprotein. Results In the Gensini score≥20 group, the proportion of diabetic patients and left ventricular ejection fraction were higher than those in the Gensini score＜20 group, the difference was statistically significant (P＜0.05). In terms of lipid-related indicators, the levels of remnant cholesterol, triglyceride and total cholesterol in Gensini score≥20 group were higher than those in the Gensini score＜20 group, the differences were statistically significant (P＜0.05). The correlation analysis of remnant cholesterol and triglyceride in Gensini score≥20 group was conducted. The results showed that there was a correlation between remnant cholesterol and triglyceride (r＝0.535, P＜0.01).Subsequently, patients with triglyceride≥1.7 mmol/L were screened for further correlation analysis, and the results showed that the correlation between remnant cholesterol and triglyceride increased further (r＝0.625, P＜0.01). Logistic regression showed that remnant cholesterol (OR＝8.0,5%CI:2.87～27.53, P＜0.01), low density lipoprotein cholesterol (OR＝3.6,5%CI:1.93～6.57, P＜0.01) and diabetes mellitus (OR＝2.4,5%CI:1.10～5.00, P＜0.05) were significantly associated with severe coronary artery stenosis. Conclusion Remnant cholesterol, low density lipoprotein cholesterol, diabetes mellitus and other factors are closely related to the occurrence of severe coronary artery stenosis in patients with prehypertension complicated with coronary heart disease.

Abstract:Aim To investigate the relationship between serum microRNA-29c (miR-29c), basic fibroblast growth factor (bFGF) levels and diabetic retinopathy (DR). Methods 154 patients with type 2 diabetes mellitus (T2DM) were selected and divided into DR group (n=65) and NDR group (n=89) according to whether they had combined retinopathy or not, and another 64 physically healthy people were selected as the control group during the same period. Spearman correlation analysis was used to correlate serum miR-29c with bFGF levels in the DR group. Multi-factor Logistic regression was used to analyze the factors influencing the occurrence of DR. ROC curve was used to analyze the diagnostic value of serum miR-29c and bFGF levels for DR. Results Serum miR-29c and bFGF levels gradually increased in the control group, NDR group and DR group (P＜0.05). Spearman correlation analysis showed that serum miR-29c was positively correlated with bFGF levels in the DR group (r_s=0.593, P＜0.001). Multi-factor Logistic regression analysis showed that disease duration (OR=1.2,5%CI:1.046 to 1.226), glycosylated hemoglobin A1c (HbA1c) (OR=2.1,5%CI:1.499 to 2.725), miR-29c (OR=2.3,5%CI:1.132 to 3.943), and bFGF (OR=3.8,5%CI:1.178 to 5.304) were independent risk factors for DR (P＜0.05). The ROC curves showed that the sensitivity and specificity of miR-29c combined with bFGF for the diagnosis of DR (76.92% and 83.15%) were higher than those of miR-29c alone (64.62% and 78.65%), bFGF (75.38% and 60.67%) diagnosis. Conclusions Serum miR-29c and bFGF levels are significantly elevated in patients with T2DM, which are closely related and jointly involved in retinopathy development. Combined detection of serum miR-29c and bFGF levels can improve the diagnostic efficacy of DR.

Abstract:Aim To investigate the effect of baseline peripheral blood circulating monocyte (MO) level on long-term mortality risk after percutaneous coronary intervention (PCI) in patients with coronary heart disease (CHD). Methods 6 045 patients with CHD treated by PCI were analyzed retrospectively. According to the level of peripheral blood circulating monocyte at baseline, the patients were divided into three groups and followed up:Ⅰ group:MO＜0.40×10⁹ L－1 (n＝1 943); Ⅱ group:0.40×10⁹ L－1≤MO≤0.56×10⁹ L－1 (n＝2 072); Ⅲ group:MO＞0.56×10⁹ L－1 (n＝2 030). The end points of follow-up included all-cause mortality (ACM), cardiac mortality (CM), major adverse cardiovascular event (MACE) and major adverse cardiac and cerebrovascular event (MACCE). The mean follow-up time was (35.9±22.6) months. Results 309 patients developed ACM, including 73 cases (3.8%) in Ⅰ group, 98 cases (4.7%) in Ⅱ group, and 138 cases (6.8%) in Ⅲ group; 251 patients developed CM, including 58 cases (3.0%) in Ⅰ group, 80 cases (3.9%) in Ⅱ group and 113 cases (5.6%) in Ⅲ group; 785 patients developed MACE, including 226 cases (11.6%) in Ⅰ group, 248 cases (12.0%) in Ⅱ group and 311 cases (15.3%) in Ⅲ group; 862 patients developed MACCE, including 250 cases (12.9%) in Ⅰ group, 269 cases (13.0%) in Ⅱ group and 343 cases (16.9%) in Ⅲ group. There were significant differences in ACM, CM, MACE and MACCE among the three groups (P≤0.001). Kaplan-Meier curve analysis of ACM and CM in the three groups showed that the prognosis of patients was worse with the increase of baseline peripheral blood circulating monocyte level (P＜0.05). COX regression analysis showed that the increase of baseline peripheral blood circulating monocyte level was independently correlated to the occurrence of ACM (Ⅲ group vs Ⅰ group, HR=1.8,5%CI:1.056-1.905, P＝0.020) and CM (Ⅲ group vs Ⅰ group, HR=1.5,5%CI:1.023-1.983, P＝0.036). Conclusion High levels of baseline peripheral blood circulating monocyte level are an independent predictor of long-term mortality risk in CHD patients after PCI.

Abstract:Aim To investigate the correlation between serum long-chain non-coding RNA GAS5 (lncRNA GAS5) level and in-stent restenosis in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods 144 patients with STEMI who received selective percutaneous coronary intervention were selected, and according to the follow-up results, the patients were divided into 27 cases of in-stent restenosis (ISR) group and 109 cases of non-ISR group, with 8 cases lost. The clinical data of patients with STEMI were collected. The level of serum lncRNA GAS5 in all patients after percutaneous coronary intervention was measured by real-time quantitative PCR (qRT-PCR). Results Compared with non-ISR group, the proportions of smokers before operation and complicated diabetic, total bilirubin (TBIL), levels of total cholesterol (TC) and low density lipoprotein cholesterol (LDLC) in serum in ISR group were increased (P＜0.05), and the level of lncRNA GAS5 in serum was decreased (P＜0.05); The incidence of ISI in low level of GAS5 was higher than that in the patients with high level of lncRNA GAS5 (P＜0.05), and the average time of no restenosis was shorter (P＜0.05); Logistic regression analysis showed that low level of lncRNA GAS5 was a risk factor of in-stent restenosis in patients with STEMI. Conclusion The level of lncRNA GAS5 is down-regulated in the serum of in-stent restenosis patients with STEMI, and it is a risk factor of in-stent restenosis patients with STEMI.

Abstract:Aim To evaluate the predictive value of tenascin-C (TNC) in coronary heart disease (CHD) and coronary atherosclerosis severity. Methods 160 patients with chest pain who had undergone coronary angiography were included. According to the results of the angiography, the patients were divided into two groups:CHD group (n＝90) and non-CHD group (n＝70). The coronary atherosclerosis severity was evaluated by Gensini score, and the CHD patients were divided into three groups:low Gensini score group (n＝39), medium Gensini score group (n＝27), high Gensini score group (n＝24). The levels of serum TNC were compared in each group. Pearson correlation analysis and multivariate linear regression analysis were used to evaluate the correlation between TNC and Gensini score. The predictive value of TNC in patients with coronary heart disease and high Gensini score was analyzed by ROC curve. ResultsThe level of serum TNC in CHD group was significantly higher than that in non-CHD group (P＜0.001), the serum TNC level of high Gensini score group was significantly higher than that of medium Gensini score group, and the serum TNC level of medium Gensini score group was significantly higher than that of low Gensini score group (P＜0.001). Pearson correlation analysis and multivariate linear regression analysis showed that there was a significant positive correlation between the serum TNC level and Gensini score in CHD group (P＜0.001). ROC curve analysis showed that the area under curve (AUC) of TNC for predicting CHD was 0.811 (95%CI:0.768-0.974), and the Cut-off value was 11.94 μg/L, the sensitivity and specificity were 83.5% and 76.4% respectively. The AUC of TNC in predicting patients with high Gensini score was 0.944 (95%CI:0.816-0.989), and the Cut-off value was 16.47 μg/L, the sensitivity and specificity were 82.6% and 78.4% respectively. Conclusion Serum TNC level is significantly correlated with Gensini score of CHD patients, which has a certain predictive value for CHD and coronary atherosclerosis severity.

Abstract:Aim To investigate the efficacy of different doses of atorvastatin in the treatment of acute coronary syndrome (ACS) patients with baseline low-level low density lipoprotein. Methods 102 patients with ACS associated with baseline low-level low density lipoprotein were selected from January 2016 to December 2017. The patients were randomly divided into three groups:low-dose short-term group, high-dose short-term group and low-dose long-term group, with 34 cases in each group. Low-dose short-term group and low-dose long-term group were treated with atorvastatin 10 mg/day for 9 months and 12 months respectively, high-dose short-term group was treated with atorvastatin 40 mg/day for 9 months.Automatic biochemical analyzer was used to detect triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDLC) and low density lipoprotein cholesterol (LDLC). The blood lipid levels and the incidence of major adverse cardiovascular and cerebrovascular events of the three groups of patients were compared. Results Before treatment, there were no significant differences in baseline data and 4 indexes of blood lipid among the three groups (P＞0.05).After 3 and 9 months of treatment, the levels of TC, LDLC and TG in the high-dose short-term group were lower than those in the low-dose short-term group and low-dose long-term group, and the levels of HDLC were higher than those in the low-dose short-term group and low-dose long-term group (all P＜0.05). After 12 months of treatment, the efficacy of 4 indexes of blood lipid in the low-dose long-term group was improved better than that in the low-dose short-term group after 9 months of treatment. There was no significant difference in 4 indexes of blood lipid in the low-dose long-term group after 12 months of treatment and high-dose short-term group after 9 months of treatment (P＞0.05). There was no death at the end of the treatment in the three groups. The incidence of major unconscience-related cerebrovascular events in the high-dose short-term group and low-dose long-term group was 11.76%, slightly lower than that in the low-dose short-term group (17.65%), but there was no statistical significance in the three groups (P＞0.05). Conclusion The use of atorvastatin in the treatment of ACS with baseline low-level low density lipoprotein is dose-dependent and time-dependent, so it is recommended to increase the dose and prolong the treatment time.

Abstract:The novel coronavirus pneumonia, also known as the coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The current clinical evidence shows that the risk of death is significantly increased in patients with COVID-19 combined with underlying cardiovascular diseases. Most patients will develop myocarditis, myocardial injury, arrhythmia and cardiomyopathy in the course of the disease. This article mainly describes the current research situation of cardiovascular complications of COVID-19.

Abstract:Betatrophin, also known as lipasin or angiopoietin-like protein 8 (ANGP TL8), is mainly expressed in human liver and adipose tissue. There is increasing evidence to show the correlation between betatrophin expression and blood lipid levels, and especially in patients of obesity or diabetes, stimulated by factors such as insulin, thyroid hormone, and energy intake, it can lead to an increase in betatrophin levels. It has also been verified in animal models that betatrophin can increase serum triglyceride levels and promote the proliferation of pancreatic β-cells. This article reviews the latest research progress of betatrophin in lipid regulation.

Abstract:Atherosclerosis is a common systemic vascular disease, and it is also the pathological basis of a variety of acute cardiovascular diseases. Acute cardiovascular disease has a high mortality rate, which seriously endangers people's life and health. In recent years, more and more studies have shown that the RhoA/ROCK signaling pathway is closely related to atherosclerosis. Inhibition of RhoA/ROCK signaling pathway can delay or inhibit the formation and pathological changes of atherosclerosis by stabilizing vascular endothelial function, inhibiting the proliferation and migration of vascular smooth muscle cells, inhibiting vascular calcification, regulating inflammatory cell aggregation, and inhibiting platelet proliferation and deformation.

Abstract:Cardiovascular diseases, as a major threat to human health,are attracting more and more attention. Although the treatment system for cardiovascular diseases has been preliminarily completed, there are still residual risks after controlling related risk factors. It is still difficult and the mortality remains stubbornly high. Intestinal flora is an important part of human physiological and metabolic homeostasis, especially its metabolite level may be closely related to some diseases. Intestinal flora metabolites are regarded as a new break-through in the prevention and treatment of cardiovascular diseases. The purpose of the current review is to explore the relationship between intestinal flora metabolites and common cardiovascular diseases such as coronary heart disease, heart failure, hypertension and arrhythmia,hoping to provide new ideas for the prevention and treatment of cardiovascular diseases.

Abstract:The low density lipoprotein receptor-related protein 6 (LRP6) is one of the important receptor proteins that perform the canonical Wnt/β-catenin pathway. Mutations in the protein are often associated with a variety of human diseases, including metabolic syndrome, tumor, Alzheimer's disease and osteoporosis. The mutation of low-density lipoprotein receptor-related protein 6 will have important guiding significance in the clinical diagnosis. In this paper, the function of low-density lipoprotein receptor-related protein 6 and the occurrence and development of related diseases will be summarized to provide useful resources for future research.