High glucose levels accelerate atherosclerosis via NLRP3-IL/ MAPK/NF-κB-related inflammation pathways.

BACKGROUND: As a chronic inflammatory disease, diabetes mellitus (DM) contributes to the development of atherosclerosis (AS). However, how the NLRP3 inflammasome participates in diabetes-related AS remains unclear. Therefore, this study aimed to elucidate the mechanism through which NLRP3 uses high glucose (HG) levels to promote AS. METHODS: Serum and coronary artery tissues were collected from coronary artery disease (CAD) patients with and without DM, respectively. The expression of NLRP3 was detected, and the effects of this inflammasome on diabetes-associated AS were evaluated using streptozotocin (STZ)-induced diabetic apoE-/- mice injected with Adenovirus-mediated NLRP3 interference (Ad-NLRP3i). To elucidate the potential mechanism involved, ox-LDL-irritated human aortic smooth muscle cells were divided into the control, high-glucose, Si-NC, and Si-NLRP3 groups to observe the changes induced by downregulating NLRP3 expression. For up-regulating NLRP3, control and plasmid contained NLRP3 were used. TNF-α, IL-1ß, IL-6, IL-18, phosphorylated and total p38, JNK, p65, and IκBα expression levels were detected following the downregulation or upregulation of NLRP3 expression. RESULTS: Patients with comorbid CAD and DM showed higher serum levels and expression of NLRP3 in the coronary artery than those with only CAD. Moreover, mice in the Ad-NLRP3i group showed markedly smaller and more stable atherosclerotic lesions compared to those in other DM groups. These mice had decreased inflammatory cytokine production and improved glucose tolerance, which demonstrated the substantial effects of NLRP3 in the progression of diabetes-associated AS. Furthermore, using the siRNA or plasmid to downregulate or upregulate NLRP3 expression in vitro altered cytokines and the MAPK/NF-κB pathway. CONCLUSIONS: NLRP3 expression was significantly increased under hyperglycemia. Additionally, it accelerated AS by promoting inflammation via the IL/MAPK/NF-κB pathway.

SFI, a sex hormone binding globulin based nomogram for predicting non-alcoholic fatty liver disease in the Chinese population.

Background: The purpose of this study is to establish a novel nomogram model for accurate detection of non-alcoholic fatty liver disease (NAFLD) in the Chinese population based on sex hormone binding globulin (SHBG) and other routine laboratory tests. Methods: A total of 1417 participants (1003 testing and 414 validations) were enrolled into the study. Risk factors independently associated with NAFLD were identified and incorporated in the new nomogram, SFI. The performance of nomogram was assessed by analysis of receiver operating characteristic (ROC) curve, calibration curve, and decision curve. Results: We formulated a new nomogram incorporating four independent factors: SHBG, body mass index (BMI), ALT/AST, and triglycerides (TG). The nomogram achieved good indexes of area under ROC 0.898 (95% confidence interval 0.865-0.926) in predicting NAFLD, which was significantly superior to previously reported models of FLI, HSI, LFS, and LAP. The calibration curve and decision curve demonstrated high performance and clinical utility of the nomogram in predicting NAFLD. Conclusion: The nomogram SFI has high performance in predicting NAFLD in Chinese population and may be used as a cost-effective screening model to assess NAFLD in the general population.

Performance of Problem-Based Learning Based Image Teaching in Clinical Emergency Teaching.

At present, with the rapid increase of emergency knowledge and the improvement of people's requirements for medical quality, the traditional teaching mode cannot fully meet the needs of emergency teaching in the new era. PBL is a project-based teaching that allows students to have a deeper understanding of content knowledge and to better apply what they have learned to their lives. This paper aims to improve the clinical emergency teaching mode by PBL teaching method, and improve the comprehensive ability of clinical emergency of medical students. This article proposes a problem-based PBL imaging teaching method, combining the characteristics and content of clinical emergency courses, focusing on students, highlighting the problem-solving process, and improving students' creative thinking ability. To cultivate students' interest in clinical learning, develop their self-learning ability, train their teamwork and communication skills, and cultivate their ability to set, question and solve questions, so as to promote medical students' overall comprehensive ability to integrate specialized knowledge and clinical practice. In this paper, the PBL teaching method and the traditional teaching method of comparative experiments show that the PBL teaching method can more effectively highlight the characteristics of clinical emergency medicine teaching mode, and make full use of the limited emergency teaching resources, so as to improve the quality of clinical emergency teaching. Compared with the traditional teaching mode, the theoretical knowledge and clinical operation skills of medical students under the PBL teaching mode are improved by 13%, Autonomous learning ability, communication ability and creative thinking ability have also been relatively improved.

Inclisiran inhibits oxidized low-density lipoprotein-induced foam cell formation in Raw264.7 macrophages via activating the PPARγ pathway.

Proprotein convertase subtilisin kexin type 9 (PCSK9) is a well-known proprotein convertase that influences foam cell formation and modulates atherosclerosis. Inclisiran is a novel chemosynthetic small interfering RNA that inhibits PCSK9 synthesis. This study aimed to explore the effect of inclisiran on oxidized low-density lipoprotein (ox-LDL)-induced foam cell formation in Raw264.7 macrophages and to investigate the underlying mechanisms. Raw264.7 cells were treated with ox-LDL to induce the formation of macrophage-derived foam cells. Oil Red O staining and high-performance liquid chromatography were performed to detect lipid accumulation and cholesterol levels. Dil-ox-LDL uptake assay, CCK-8, RT-qPCR, and Western blotting analysis were performed to examine ox-LDL uptake, cell viability, and expression of scavenger receptor-related factors. Inclisiran reduced lipid accumulation in ox-LDL-treated macrophages in a dose-dependent manner. Inclisiran significantly inhibited the levels of total cholesterol, free cholesterol, and cholesterol ester in the supernatant of Raw264.7 cells. Inclisiran reduced ox-LDL uptake and increased Raw264.7 cell viability. Meanwhile, inclisiran downregulated the expression of SR-A, LOX-1, and CD36 and upregulated SR-BI, ApoE, and ABCA1. Furthermore, inclisiran increased PPARγ activity and decreased NF-κB activity. An inhibitor of PPARγ (T0070907) reversed the beneficial effects of inclisiran on ox-LDL uptake, NF-κB inactivation, and cytokine expression. In conclusion, these data suggested that inclisiran inhibited the formation of macrophage-derived foam cells by activating the PPARγ pathway.HighlightsInclisiran reduces lipid accumulation in Raw264.7 cells;Inclisiran reduces ox-LDL uptake and increases Raw264.7 cell viability;Inclisiran inhibits foam cell formation by activating the PPARγ pathway.

Melatonin protected against myocardial infarction injury in rats through a Sirt6-dependent antioxidant pathway.

BACKGROUND: The Sirt6, one of the members of the sirtuin family, has been regarded as a key factor in the pathogenesis of myocardial infarction (MI) through its antioxidant defense mechanisms. A previous study reported that melatonin is an antioxidant drug that can act as an agent for cardioprotection in cardiac ischemia-reperfusion (I/R) injury. However, whether melatonin could protect against cardiac remodeling after myocardial injury via the Sirt6-dependent antioxidant pathway remains unknown. OBJECTIVES: To explore the protective effects and the potential mechanisms of melatonin on MI-induced injury in rats. MATERIAL AND METHODS: A cardiac remodeling model was established through left coronary artery ligation surgery. The dose of melatonin was 10 mg/kg body weight. Four weeks after the treatment for 7 successive days, the infarct size and hemodynamic parameters were evaluated. The relative mRNA level and protein level of Sirt6 were also determined. Finally, the levels of oxidative stress, including reactive oxygen species (ROS) and superoxide dismutase (SOD), were measured, and the expression of nitric oxide (NO), inducible nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS) and their corresponding phosphorylation were evaluated. RESULTS: After the treatment with melatonin, infarct size, the left ventricular end-diastolic diameter (LVEDd), and left ventricular end-systolic diameter (LVEDd) and minimum first derivative of developed pressure (min dP/dt) decreased, while left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS) and maximum first derivative of developed pressure (max dP/dt) increased in the melatonin-MI (MM) group compared to the placebo-MI (PM) group. Furthermore, the expressions of Sirt6, both in mRNA and protein level, were significantly increased in the MM group treated with melatonin, as compared to the melatonin-control (MC) group treated with melatonin. In addition, melatonin enhanced SOD activity and reduced ROS levels. At the same time, we observed that the eNOS/NO signaling pathways were activated. CONCLUSIONS: Melatonin improved cardiac function through the Sirt6-dependent antioxidant pathway in MI rats.

Transforming Growth Factor ß1 (TGF-ß1) Appears to Promote Coronary Artery Disease by Upregulating Sphingosine Kinase 1 (SPHK1) and Further Upregulating Its Downstream TIMP-1.

BACKGROUND Transforming growth factor (TGF)-ß1 is involved in the pathogenesis of coronary artery disease (CAD), but the mechanism of its action remains unclear. Our study aimed to investigate the role of TGF-ß1 in CAD and to explore the possible mechanisms. MATERIAL AND METHODS A total of 60 CAD patients and 54 healthy people were included in this study. Blood samples were drawn from each participant to prepare serum. ELISA was utilized to measure serum level of TGF-ß1. TGF-ß1 expression vector, TGF-ß1 siRNA, and TIMP-1 siRNA were transfected into human primary coronary artery endothelial cell (HCAEC) line cells, and expression of TGF-ß1 sphingosine kinase 1 (SPHK1) and TIMP metallopeptidase inhibitor 1 (TIMP-1) was detected by Western blot. Cell apoptosis was detected by MTT assay. RESULTS Serum level of TGF-ß1 was specifically higher in patients with CAD than in healthy controls. Serum levels of active TGF-ß1 can be used to effectively distinguish CAD patients from healthy controls. TGF-ß1 overexpression promoted the apoptosis of HCAEC and TGF-ß1 siRNA silencing inhibited the apoptosis of HCAEC. TGF-ß1 overexpression also promoted the expression of SPHK1 and TIMP-1. SPHK1 overexpression upregulated TIMP-1 but it showed no significant effects on TGF-ß1. TIMP-1 overexpression showed no significant effects on TGF-ß1 or SPHK1. SPHK1 inhibitor and TIMP-1 silencing reduced the enhancing effects of TGF-ß1 overexpression on cell apoptosis. CONCLUSIONS TGF-ß1 appears to promote CAD through the induction of cell apoptosis by upregulating SPHK1 expression and further upregulating its downstream TIMP-1.

MicroRNA­221­3p contributes to cardiomyocyte injury in H2O2­treated H9c2 cells and a rat model of myocardial ischemia­reperfusion by targeting p57.

Myocardial ischemia­reperfusion (I/R) injury is a major cause of cardiovascular disease worldwide, and microRNAs have been implicated in the regulation of pathological and physiological processes in myocardial I/R injury. The present study aimed to investigate the role of microRNA (miR)­221­3p in myocardial I/R injury. Cell death and lactate dehydrogenase (LDH) activity were increased in hydrogen peroxide (H2O2)­treated H9c2 cells, as measured by flow cytometry and an LDH detection kit. The expression of miR­221­3p was elevated in H2O2­incubated cells and in remote areas of the rat I/R model, examined using reverse transcription­quantitative polymerase chain reaction analysis. The overexpression of miR­221­3p enhanced the number of propidium iodide (PI)+ cells and the activity of LDH in H2O2­treated cells. In I/R­induced rats, the overexpression of miR­221­3p promoted the number of myosin+ cells and inhibited the fractional shortening of left ventricular diameter (FSLVD%). The results showed that the expression of p57 at the gene and protein levels was decreased in H9c2 cells incubated with H2O2 and in rats subjected to I/R surgery; the expression of p57 decreased following the overexpression of miR­221­3p. Subsequently, the hypothesis that p57 was the direct target of miR­221­3p was confirmed by performing a dual­luciferase reporter assay. Finally, to examine the function of p57 in myocardial impairment, p57 was transfected into H9c2 cells and administered to the rats prior to undergoing H2O2 treatment and I/R surgery, respectively. The results indicated that p57 attenuated the number of PI+ cells and the activity of LDH in H2O2­treated cells, whereas p57 downregulated the number of myosin+ cells and upregulated FSLVD% in the I/R­treated rats. Therefore, these findings suggested that miR­221­3p exacerbated the H2O2­induced myocardial damage in H9c2 cells and myocardial I/R injury in the rat model by modulating p57.

Relationship between carotid artery sclerosis and blood pressure variability in essential hypertension patients.

OBJECTIVES: This study aimed to investigate the relationship between the presence of carotid arteriosclerosis (CAS) and blood pressure variability (BPV) in patients with essential hypertension. METHODS: One hundred and forty four essential hypertension patients underwent ambulatory BP monitoring for 24h after hospitalization. Common BPV metrics were calculated. General clinical parameters, including age, gender, height, weight, history of coronary heart disease, stroke, diabetes, hypertension, smoking and drink, were recorded. Biochemical indices were obtained from a blood test. Carotid intima-media thickness (IMT) and carotid plaques were assessed to separate patients into a non-CAS group (IMT≤0.9mm; n=82) and a CAS group (IMT>0.9mm; n=62). BPV metrics and clinical parameters were analyzed and compared between the two groups. Multivariate logistic regression analysis was performed to determine the associated risk factors of CAS. RESULTS: Multivariate logistic regression analysis revealed that two BPV metrics, the standard deviation of daytime systolic blood pressure (SSD) (OR: 1.587, 95%CI: 1.242-2.028), the difference between average daytime SBP and nighttime SBP (OR: 0.914, 95%CI: 0.855-0.977), as well as three clinical parameters (age, OR: 1.098, 95%CI: 1.034-1.167; smoking, OR: 4.072, 95%CI: 1.466-11.310, and fasting blood glucose, OR: 2.029, 95%CI: 1.407-2.928), were significant factors of CAS in essential hypertension patients. CONCLUSION: SSD, in combination with the ageing, smoking and FBG, has been identified as risk factors for CAS in patients with essential hypertension.

MiR-139-5p protect against myocardial ischemia and reperfusion (I/R) injury by targeting autophagy-related 4D and inhibiting AMPK/mTOR/ULK1 pathway.

This study aimed at investigating the effect and underlying mechanism of miR-139-5p in myocardial ischemia and reperfusion (I/R) injury. A hypoxia/ reoxygenation (H/R) model was established in H9c2 cardiomyocytes. The level of miR-139-5p was detected in H/R-treated cardiomyocytes, and subsequently, the level of miR-139-5p or its target gene autophagy-related 4D (ATG4D) was up- or downregulated. Furthermore, the cell viability, apoptosis, and autophagy, as well as the expression levels of the proteins related to adenosine 5'-monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)/uncoordinated 51-like kinase 1 (ULK1) signaling pathway were determined. The MiR-139-5p was downregulated in H/R-treated cardiomyocytes in comparison to the untreated cells (P < 0.05). H/R treatment significantly decreased the cell viability but increased the cell apoptosis ratio, and autophagy-related proteins levels (P < 0.05). The overexpression of MiR-139-5p significantly promoted cell apoptosis and inhibited cell autophagy induced by H/R (P < 0.05); however, the effects of miR-139-5p on cell apoptosis and cell autophagy were inhibited by its target gene ATG4D (P < 0.05). Furthermore, the upregulated miR-139-5p remarkably inhibited the expression of p-AMPK, p-Raptor, and ULK1, but increased that of p-mTOR (P < 0.05) in H/R-treated cardiomyocytes. The MiR-139-5p has the potential of regulating cell apoptosis and cell autophagy by inhibiting AMPK/mTOR/ULK1 signaling pathway and thereby protecting against myocardial I/R injury.

BRD7 mediates hyperglycaemia-induced myocardial apoptosis via endoplasmic reticulum stress signalling pathway.

Bromodomain-containing protein 7 (BRD7) is a tumour suppressor that is known to regulate many pathological processes including cell growth, apoptosis and cell cycle. Endoplasmic reticulum (ER) stress-induced apoptosis plays a key role in diabetic cardiomyopathy (DCM). However, the molecular mechanism of hyperglycaemia-induced myocardial apoptosis is still unclear. We intended to determine the role of BRD7 in high glucose (HG)-induced apoptosis of cardiomyocytes. In vivo, we established a type 1 diabetic rat model by injecting a high-dose streptozotocin (STZ), and lentivirus-mediated short hairpin RNA (shRNA) was used to inhibit BRD7 expression. Rats with DCM exhibited severe myocardial remodelling, fibrosis, left ventricular dysfunction and myocardial apoptosis. The expression of BRD7 was up-regulated in the heart of diabetic rats, and inhibition of BRD7 had beneficial effects against diabetes-induced heart damage. In vitro, H9c2 cardiomyoblasts was used to investigate the mechanism of BRD7 in HG-induced apoptosis. Treating H9c2 cardiomyoblasts with HG elevated the level of BRD7 via activation of extracellular signal-regulated kinase 1/2 (ERK1/2) and increased ER stress-induced apoptosis by detecting spliced/active X-box binding protein 1 (XBP-1s) and C/EBP homologous protein (CHOP). Furthermore, down-regulation of BRD7 attenuated HG-induced expression of CHOP via inhibiting nuclear translocation of XBP-1s without affecting the total expression of XBP-1s. In conclusion, inhibition of BRD7 appeared to protect against hyperglycaemia-induced cardiomyocyte apoptosis by inhibiting ER stress signalling pathway.

Gene environment interaction of GALNT2 and APOE gene with hypertension in the Chinese Han Population.

In some GWAs studies, GALNT2 and APOE polymorphisms have been identified to be related to alterations of plasma or serum HDL-C and TG concentrations. The purpose of our study is to assess the contribution of GALNT2 rs4846914, APOE rs429358, rs7412, rs405509 variants, and several environmental factors to the development of hypertension disease in the China Han population. A hospital-based case-control study was conducted. Cases were hypertension (n=211) and controls were normal participants (n=434). The AA, AG, and GG genotype frequencies of GALNT2 rs4846914 were 22.8%, 43.1%, and 34.1% in hypertension subjects, and 35.3%, 44.2%, and 20.5% in controls (P<0.05), respectively. The OR of the AG genotype adjusted for all risk factors compared to the AA genotype was 1.61 (95%CI: 1.02 to 2.56) and to the GG genotype 2.67 (95%CI: 1.59 to 4.488). There was no significant difference between the APOE rs429358, rs7412, and rs405509 genotype frequencies in hypertension and control subjects. The present work indicates that SNP rs4846914 in GALNT2 gene is related to an increased risk of hypertension in China Han population, but the APOE gene is not.