Abstract:Vascular calcification is highly prevalent in ageing, atherosclerosis, diabetes and chronic kidney disease, and associated with morbidity and mortality of myocardial infarction. As more investigation input in this field, the better understanding of vascular calcification has been achieved. This review focuses on the molecular mechanism of vascular calcification, including osteogenic transition of vascular smooth muscle cell, defects in calcification inhibitors, disorders of calcium and phosphate homeostasis, oxidative stress, inflammation, apoptosis, autophagy, extracellular matrix remodeling and microRNAs. In this column, several research groups have conducted the in-depth studies on the role of activated T-cell cytoplasmic 1 in the progression of diabetic vascular calcification, the relationship between the ratio of neutrophils to lymphocytes and coronary calcification in dialysis patients, and the effect of Dendrobium huoshanense on atherosclerosis and vascular calcification in LDLR gene knockout mice induced by high-fat food feeding.

Abstract:Aim To study the effect of Dendrobium Huoshanense C.Z. Tang et S.J.Cheng(DH)on the development of atherosclerosis and vascular calcification induced by high-fat diet in LDLR－/－ mice and the possible mechanisms.Methods Male LDLR－/－ mice were randomly divided into 2 groups and received the following treatment:High-fat diet (control group); HFD containing DH (DH group). The treatment was lasted for 18 weeks. At the end of treatment, all the mice were anesthetized followed by collection of blood, aorta and liver samples. The serum was prepared followed by determination of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDLC) and low density lipoprotein cholesterol (LDLC) levels. Whole aorta and frozen cross sections of aortic root were prepared, and the lesions in en face aorta and aortic root cross sections were determined by oil red O staining. Frozen cross sections of aortic root were also stained by Sirius Red and Alizarin Red S to determine the plaque stability and vascular calcification. Liver frozen sections were prepared and used to determine hepatic lipid content by oil red O staining. Results After 18 weeks, DH lowered serum TG to (2.31±0.16) mmol/L compared to the control group (3.76±0.47) mmol/L(P＜0.05), but had no effect on transaminase level and otherlipid profilesin serum. Meanwhile, DH significantly reduced lesions in en face aorta and sinus lesion of aortic root by 20% (P＜0.05). It also increased collagen content by 70% in plaques (P＜0.05), promoted lesion stability and reduced necrotic cores in arterial wall. At last, Alizarin Red S staining showed that the development of vascular calcified area, which was 0.88% in DH group while 1.34% in control group, was attenuated by 34% (P＜0.01). Conclusion DH inhibits the development of atherosclerosis and vascular calcification.

Abstract:Aim To explore the role of activated T-cell cytoplasmic 1 (NFATc1) in diabetic vascular calcification. Methods Fifteen patients with diabetic foot amputation were enrolled. Serum and anterior tibial arteries were collected. According to the calcium content of the anterior tibial arteries, patients were divided into low-calcification group and high-calcification group. The NFATc1 levels in serum and anterior tibial arteries were detected, and their correlation with the calcium content of anterior tibial arteries was analyzed respectively. An in vitro model of diabetic vascular calcification was constructed with mouse aortic smooth muscle cell (MASMC)to detect the phenotypic transition and calcium deposition of MASMC under high glucose. Furthermore, siRNA was used to silence NFATc1 to determine the effect of NFATc1 on MASMC phenotype transition and calcium deposition. Results The NFATc1 level of the high-calcification group was significantly higher than that in the low-calcification group both in serum and in anterior tibial arteries. And correlation analysis showed that the serum and anterior tibial artery NFATc1 levels were positively correlated with calcium content. In the in vitro model of MASMC, high glucose significantly weakened the contraction phenotype of MASMC, promoted the osteogenic phenotype transdifferentiation, and significantly increased the calcium deposition. The effect of high glucose in promoting phenotypic transition and promoting calcification was not connected with its hypertonicity. After silencing NFATc1 with siRNA, the osteogenic differentiation of MASMC was significantly inhibited, and calcium deposition was also significantly reduced. Conclusion NFATc1 can promote the transdifferentiation of vascular smooth muscle cell to an osteogenic phenotype and promote the progression of diabetic vascular calcification.

Abstract:Aim To explore the value of neutrophil/lymphocyte ratio (NLR) in predicting CAC by analyzing the risk factors of coronary artery calcification (CAC) in dialysis patients. Methods 163 dialysis patients (including 102 hemodialysis patients and 61 peritoneal dialysis patients) were retrospectively studied using a cross-sectional survey method. Based on the Agatston’s coronary calcification score (CACS), the patients were divided into two groups:non-calcification group (CACS 0~10), and calcification group (CACS≥11). NLR, age, dialysis history, levels of high sensitivity C-reactive protein (hs-CRP), calcium, phosphorus, total parathyroid hormone (iPTH), albumin (Alb), hemoglobin, and serum creatinine were calculated and compared between the two groups. Spearman correlation was used to analyze the factors related to CAC, binary Logistic regression analysis was used to evaluate the risk factors of CAC, and the receiver operating characteristic (ROC) curve was used to explore the predictive value of NLR for CAC. Results The total detection rate of CAC in 163 dialysis patients was 63.8%. NLR was higher in calcification group (n＝104) than that in non-calcification group (n＝59) (P＜0.001). Patients with calcification were divided into mild calcification group(CACS 11~400)and severe calcification group (CACS＞400), there was no difference in NLR between the two subgroups. Spearman correlation analysis showed that the NLR was significantly correlated with CAC (r＝0.403, P＜0.001). Binary Logistic regression analysis showed that age (OR=1.069, P＜0.001), dialysis age (OR=1.024, P＜0.001), diabetes (OR=15.871, P＝0.012) and NLR (OR=1.720, P＝0.001) were risk factors for CAC. ROC curve analysis results showed that when using the combined index of NLR and age to predict CAC in dialysis patients, the area under the curve was 0.810 (95%CI 0.739~0.880, P＜0.001), which was significantly higher than NLR (0.2,5%CI 0.666~0.818, P＜0.001) and age (0.4,5%CI 0.674~0.834, P＜0.001) when analyzed separately under the curve. Conclusion Dialysis patients with advanced age and high levels of NLR have a higher risk of CAC, and the combination of NLR and age has a better predictive value for the occurrence of CAC.

Abstract:Vascular calcification refers to the deposition of hydroxyapatite in the vascular wall, which is involved and regulated by multiple factors, similar to the active biological process of bone and cartilage formation. Endoplasmic reticulum stress is an adaptive regulatory response in the the organism. Appropriate endoplasmic reticulum stress helps maintain endoplasmic reticulum homeostasis, but excessive endoplasmic reticulum stress will promote the initiation and progression of vascular calcification. This paper reviews the potential role and molecular mechanism of endoplasmic reticulum stress, especially unfolded protein response, in vascular calcification, hoping to provide new ideas for the prevention and treatment of vascular calcification.

Abstract:Vascular calcification (VC) increases the incidence of cardiovascular events in patients with chronic kidney disease, hypertension and diabetes mellitus. However, up to now, the molecular mechanism leading to VC has not been fully elucidated, and there is a lack of effective treatment methods, so it is necessary to find new therapeutic targets.With the development of molecular biological analysis technology, non-coding RNA has become an important research hotspot. It has been reported that non-coding RNA can regulate the expressions of osteogenic gene or osteogenic inhibition gene, or regulate autophagy and aging process to regulate VC under the stimulation of VC promoting factors. This review mainly summarizes the research progress of non-coding RNA (microRNA, long non-coding RNA and circular RNA) in VC.

Abstract:Aim The purpose of this study was to observe the effect and mechanism of angiotensin(1-7)(Ang(1-7)) on angiotensin Ⅱ(AngⅡ)-induced senescence of human umbilical vein endothelial cell(HUVEC). MethodsHUVEC were cultured in vitro with DMEM high glucose medium containing 10%fetal bovine serum (FBS) for 48 hours and randomly divided into control group, Ang(1-7) group (1 μmol/L), AngⅡ group (1 μmol/L), and AngⅡ＋Ang(1-7) group. Cell senescence β-galactosidase(SA-β-Gal) staining kits were used to detect the number of senescent cell in each group (observed under an optical microscope); reactive oxygen species (ROS) levels were measured using reactive oxygen detection kits, and p53 and dynamin related protein-1(Drp1) expression in each group were detected by Western blot.Results Compared with the control group, both SA-β-Galpositive cell and ROS levels in the AngⅡ group were significantly increased(P＜0.001), and p53 and Drp1 protein expression in the cell were significantly increased (P＜0.01). Compared with AngⅡ group, SA-β-Galpositive cell(P＜0.01), ROS levels(P＜0.001), p53 and Drp1 protein expression(P＜0.05) were decreased in AngⅡ＋Ang(1-7) group. Conclusion Ang(1-7) may inhibit the ROS generation in HUVEC by affecting the p53/Drp1 pathway, and reverse the aging of HUVEC induced by AngⅡ.

Abstract:Aim To explore possible action mechanism of mitogen-activated protein kinase (MAPK) signal transduction pathway on chemotaxis of cardiac fibroblasts (CF) induced by transforming growth factor-β1 (TGF-β1). Methods The CF of neonatal SD rats were cultured. Cell were randomly divided into blank control group, TGF-β1 group, c-Jun N-terminal kinase (JNK) inhibitor (SP600125 10 μmol/L) group, P38MAPK inhibitor (SB203580 10 μmol/L) group and extracellular signal-regulated protein kinase (ERK) inhibitor (U0126 10 μmol/L) group. Except blank control group, the other groups were given 10 μg/L TGF-β1 and corresponding inhibitors. Methyl thiazolyl tetrazolium (MTT) colorimetric assay was applied to detect cell viability of CF. Transwell chamber was applied to detect motor ability of CF. The collagen content was detected by hydroxyproline kit. The enzyme-linked immunosorbent assay (ELISA) was applied to detect levels of monocyte chemotactic protein-1 (MCP-1) and plasminogen activator inhibitor-1 (PAI-1) in CF. Western blot was applied to detect expression levels of α-smooth muscle actin (α-SMA), type Ⅰ collagen (Col-1) and matrix metalloproteinase-9 (MMP-9) in CF. Results TGF-β1 could significantly promote cell viability and chemotaxis of CF and collagen content. MAPK inhibitors could inhibit the proliferation and chemotaxis, and decrease collagen content (P＜0.05). ELISA results showed that MAPK signaling pathway inhibitors could significantly decrease increased MCP-1 and PAI-1 induced by TGF-β1 treatment (P＜0.05). Western blot results showed that MAPK signaling pathway inhibitors could significantly decrease increased expression levels of α-SMA, Col-1 and MMP-9 protein induced by TGF-β1 treatment (P＜0.05). Conclusion TGF-β1 may promote chemotaxis of CF by activating MAPK signaling pathway. The application of MAPK inhibitors can inhibit myocardial fibrosis to certain extent.

Abstract:Aim To investigate the pharmacology mechanism of Sanhuang Xiexin decoction in the treatment of atherosclerosis. Methods All the chemical components and the targets related to Sanhuang Xiexin decoction were searched by the traditional Chinese medicine system pharmacology platform (TCMSP), the oral bioavailability (OB) ≥30% and drug likeness(DL) ≥ 0.18 were used as the screening conditions for molecular compounds and the comparative Toxicogenomics Database (CTD) were used to screen the genes related to atherosclerosis. The network map was constructed by Cytoscape 3.6.1 soft ware and the DAVID database was used for pathway annotation and analysis. ResultsThe compounds-targets-pathway network of Sanhuang Xiexin decoction related to atherosclerosis contained 41 compounds, 22 corresponding targets and 39 signaling pathway. The top four compounds were quercetin, baicalein, wogonin and emodin. The top five targets were prostaglandin G/H synthase 2 (PTGS2), intercellular adhesion molecule 1 (ICAM-1), matrix metalloproteinase-9 (MMP-9), tumor necrosis factor (TNF) and interleukin-6(IL-6). Related pathway were HIF-1 signaling pathway, cytokine-cytokine receptor interaction, NF-κB signaling pathway, VEGF signaling pathway, arachidonic acid metabolism and PPAR signaling pathway. Conclusion The active components of Sanhuang Xiexin decoction may regulate arachidonic acid metabolism, NF-κB signaling pathway, VEGF signaling pathway in the treatment of atherosclerosis mainly through PTGS2, ICAM-1, MMP-9, IL-6 and other targets.

Abstract:Aim To investigate whether Nuciferine affect the proliferation of arterial endothelial cell by regulating SDF-1/CXCR4 signaling pathway. Methods Human aortic endothelial cell (HAEC) were co-cultured with Nuciferine of different concentrations (0 mg/L, 10 mg/L, 20 mg/L, 50 mg/L, 100 mg/L) for 24 hours. The expressions of SDF-1, CXCR4 mRNA and their proteins in each group were detected by RT-QPCR and Western blot techniques, respectively. The effects of different concentrations of Nuciferine on the colonization ability of HAEC were observed by endothelial cell tubule formation experiment in vitro. Results The expression of SDF-1/CXCR4 protein and mRNA increased in Nuciferine group, especially in 50 mg/L group. The tubule formation energy of HAEC in vitro was enhanced in a concentration-dependent manner in the Nuciferine group, which was the most obvious in the 50 mg/L group. Conclusion Nuciferine can promote the proliferation of vascular endothelial cell by up-regulating the expression of SDF-1/CXCR4 signaling pathway.

Abstract:Aim To investigate the changes of peripheral blood mean platelet volume (MPV), serum vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9) in patients with acute coronary syndrome (ACS) before and after percutaneous coronary intervention (PCI) and their correlation with prognosis. Methods The data of 96 patients with ACS in Geriatrics Hospital of Hainan were retrospectively analyzed. 15 patients with major adverse cardiovascular events (MACE) 6 months after PCI were included in the MACE group, and 81 patients without MACE were included in the non-MACE group. The levels of MPV of peripheral blood, serum VEGF and MMP-9 were compared between the two groups before and after PCI. Receiver operating characteristic curve (ROC) was used to analyze the predictive value of MPV of peripheral blood, serum VEGF and MMP-9 for MACE after PCI in ACS patients. The factors influencing prognosis of ACS patients were discussed. Results The levels of MPV of peripheral blood, serum VEGF and MMP-9 in ACS patients at 3 days and 7 days after PCI were lower than those before PCI (P＜0.05). The above indexes of MACE group before PCI, 3 days and 7 days after PCI were higher than those in non-MACE group (P＜0.05). ROC analysis showed that the area under curve (AUC) of MPV at 7 days after PCI was 0.987, which was larger than those before PCI and 3 days after PCI; When the cut-off value was greater than 11.27 fL, the sensitivity and specificity of MPV in predicting MACE in ACS patients undergoing PCI were 96.67% and 96.20% respectively. The AUC of VEGF was 0.906 at 7 days after PCI; When the cut-off value was greater than 173.80 ng/L, the sensitivity and specificity of VEGF in predicting MACE were 83.33% and 88.61% respectively. The AUC of MMP-9 was 0.843 at 7 days after PCI; When the cut-off value was greater than 334.74 μg/L, the sensitivity and specificity of MMP-9 in predicting MACE were 73.33% and 87.34% respectively. Conclusions The levels of MPV of peripheral blood, serum VEGF and MMP-9 are significantly decreased in ACS patients after PCI. MPV, VEGF and MMP-9 are important risk factors of MACE in ACS patients undergoing PCI, which have high clinical value in predicting MACE.

Abstract:Aim To explore the predictive value of fractional flow reserve (FFR) immediately after percutaneous coronary intervention (PCI) on the prognosis of patients. Methods This study included patients (200 cases) who underwent PCI treatment in our hospital from March 2014 to December 2017. A 1-year follow-up (re-examination by telephone and outpatient) was carried out after the operation, and the patients were divided into MACE group and non-MACE group according to whether there were major adverse cardiovascular events (MACE) during the follow-up period. Perform univariate and multiple Logistic regression analysis to analyze the influencing factors of MACE after PCI, establish receiver operating characteristic curve (ROC curve) to analyze the predictive value of FFR and draw Kaplan-Meier curve to verifyit, and analyze the influencing factors of postoperative FFR. Results After 1 year of follow-up, 31 cases were in the MACE group and 169 cases were in the non-MACE group. The postoperative FFR of MACE group was significantly lower than that of non-MACE group [t＝6.316, P＝0.000]. The proportion of patients with diabetes, hypertension and hyperlipidemia in MACE group was significantly higher than that of non-MACE group (P＜0.05). Multivariate analysis showed that hypertension, diabetes, hyperlipidemia, and postoperative FFR were the influencing factors of MACE. ROC curve showed that postoperative FFR had a better predictive value for MACE (P＝0.000), and postoperative FFR＝0.868 was the best critical value. Survival analysis showed that the incidence of MACE in the postoperative FFR≥0.868 group was later than that in the postoperative FFR＜0.868 group (Log-rank P＝0.008); multivariate analysis showed that postoperative FFR influencing factors included right coronary target vessel involvement and preoperative FFR value and stent diameter (P＝0.2,0.4,0.038, all P＜0.05). Conclusions Postoperative FFR has a good predictive value for the prognosis of PCI. The incidence of MACE in the postoperative FFR≥0.868 group is later than that in the postoperative FFR＜0.868 group. Involvement of the right coronary target vessel, preoperative FFR value and stent diameter are the influencing factors of postoperative FFR.

Abstract:With the aging of population, the burden of medical is aggravated. As a new treatment method, stem cell transplantation has reversed aging and its accompanying dysfunction. Mesenchymal stem cell, as a type of stem cell with both regeneration and immunoregulatory functions, can promote the regeneration and repair of aging tissues through direct differentiation of cell replacement and cell empowerment. At the same time, recent studies have found that continuous centrifugation can separate exosomes which is the main therapeutic carrier from mesenchymal stem cell. Senescence-related miRNA in exosomes is helpful to elucidate the molecular mechanism of anti-aging treatment of mesenchymal stem cell.

Abstract:m⁶A methyladenosine is a methylation modification catalyzed by methyltransferase at the 6th position nitrogen atom of adenine (A), which is the most common chemical modification on eukaryotic messenger RNA (mRNA). Recently, m⁶A has been found to play an important role in the dynamic regulation of RNA, which is crucial for the basic physiological processes such as adipogenesis, cell differentiation and immune/inflammatory response. Metabolic diseases are series of disorders caused by metabolic dysfunction of proteins, glucose and lipid in the body. The studies have shown that m⁶A methylation plays a key role in the development of metabolic diseases such as obesity, type 2 diabetes and cardiovascular diseases via regulation of glucose/lipid metabolism and immune/inflammation. This paper will review the role of m⁶A methylation in metabolic diseases, which may provide new ideas for the prevention and treatment of metabolic diseases from the perspective of epitranscriptome.

Abstract:Coronary artery is the earliest and most common place where atherosclerosis occurs. Serious coronary atherosclerosis can cause myocardial ischemia and myocardial infarction, which is very harmful to people's health. Compared with other arteries, coronary artery has unique anatomical structure and hemodynamic characteristics, which are significantly related to the occurrence of atherosclerosis. This article reviews the relationship between biomechanical factors and coronary atherosclerosis.