BACKGROUND:Sarcopenia is a novel complication of type 2 diabetes mellitus (T2DM), the incidence of which is rapidly increasing. Therefore, early detection and diagnosis of diabetic sarcopenia is important to improve the quality of life of patients. METHODS:Differentially expressed genes (DEGs) were identified from Gene Expression Omnibus (GEO) datasets GSE76895 and GSE1428. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were conducted using DAVID and KOBAS. Hub gene of metabolic pathways were validated using gene expression and ROC curves. Hub gene expression was then validated in mouse models and human populations, and therapeutic potential was assessed. RESULTS:The metabolic pathway is an important pathophysiological feature of T2DM and sarcopenia, with 17 genes co-enriched in this pathway. We focused on the PGK1, stained by immunofluorescence and western blot analysis, revealed significantly lower expression of PGK1 in pancreatic islets and skeletal muscles of T2DM mice, with a negative correlation with the diabetic cycle. Similarly, serum levels of PGK1 were lower in T2DM patients than in healthy individuals. Overexpression of PGK1 alleviates metabolic stress and improves skeletal muscle function in diabetic sarcopenia, highlighting its potential as a therapeutic target. CONCLUSION:These results suggest that PGK1 may serve as a novel biomarker and potential target for diabetic sarcopenia.

BACKGROUND:Periventricular nodular heterotopia 9 (PVNH9) arises from defective neuronal migration, which leads to the accumulation of ectopic neurons near the lateral ventricle. PVNH9 is caused by a heterozygous mutation in the microtubule-associated protein 1B (MAP1B) gene. METHODS:We report a PVNH9 family with a MAP1B variant. Trio whole-exome sequencing (WES) was performed to investigate the underlying genetic defects of the family. In vitro functional experiments, including messenger RNA (mRNA) splicing validation, quantitative polymerase chain reaction (qPCR) and western-blotting, were performed to determine the pathogenicity of the novel variant of MAP1B. RESULTS:WES showed that both the patient and her father carried a heterozygous MAP1B variant (c.7091 dup). Sanger sequencing confirmed that this variant occurred de novo in the father. In vitro functional experiments, mRNA splicing verification indicated normal splicing for both wild-type and mutant types. The qPCR and western blot analyses demonstrated significantly reduced mRNA and protein expression, respectively, in the mutant compared with that in the wild-type. Additionally, by comprehensive summary on genotype-phenotype associations of MAP1B in previously reported literatures, we found that loss-of-function (LOF) variants in MAP1B mainly lead to PVNH-related neurological symptoms, while patients with missense variants may only present with deafness. CONCLUSIONS:We clarified the genetic diagnosis for the family. Additionally, this study reveals a novel LOF variant in MAP1B and provides a comprehensive overview of MAP1B genotype and phenotype, aiding in the diagnosis and genetic counseling.

A comprehensive assessment of combined modifiable risk factors with common complications of type 2 diabetes (T2D) is lacking, and the potential role of proteomics remains unclear. Here, we examine the associations of cardiovascular health (CVH) score and degree of risk factor control with common diabetic complications using data from the UK Biobank (n = 14,102). Furthermore, we explore the mediation effects of plasma proteomics in a subset with proteomic data (n = 1287). Over median follow-ups of 12.4–13.4 years, higher CVH score and higher degree of risk factor control are associated with lower risks of 30 and 22 of 45 adverse outcomes among individuals with T2D, respectively. Mediation analyses reveal that mortality and multiple vascular diseases share common mediators, such as uromodulin and pro-adrenomedullin. These findings highlight the importance of risk factors modification in reducing disease burden among people with T2D and facilitate the understanding of mediation effects of plasma proteins underlying these associations. Risk factors modification is crucial for diabetes care. Here, the authors show the relationships of combined modifiable risk factors with common complications of type 2 diabetes and provide further insights into the mediating role of plasma proteomics.

BACKGROUND:Left ventricular mechanical dispersion (LV-MD), an indicator of LV dyssynchrony, is not well studied in heart failure with reduced ejection fraction (HFrEF). This study aims to investigate the profile of LV-MD in HFrEF, and evaluate whether it provides predictive information in terms of LV reverse remodeling (LVRR), particularly, on top of LV global longitudinal strain (LV-GLS). METHODS:In this observational cohort study, 239 consecutive patients hospitalized for HFrEF from 2020 to 2022 were included, and baseline and follow-up echocardiographic studies were performed. By strain analysis, LV-MD was calculated as the SD of time to peak negative longitudinal strain from 17 LV segments. LVRR was defined as an increase in LVEF of ≥10% and a decrease in LV end-diastolic diameter index of ≥10% or ≤33 mm/m2. RESULTS:During a median follow-up of 19 months, LVRR occurred in 152 patients (64%). A great heterogeneity of LV-MD ranging from 30 to 202 milliseconds was present in the cohort and 94 (84%) in 112 patients who were with LV-MD >76 milliseconds had no left bundle-branch block. LV-MD ≤76 milliseconds independently predicted LVRR and possessed greater discriminatory ability than LV-GLS by receiver operating characteristics analysis. Moreover, addition of LV-MD ≤76 milliseconds and LV-GLS ≤-5.5% to a multivariable model, including age, HF duration, systolic blood pressure, QRS duration and use of angiotensin receptor-neprilysin inhibitors, improved predictive performance in terms of LVRR. CONCLUSIONS:LV mechanical dyssynchrony is widely present in HFrEF, irrespective of left bundle-branch block. LV-MD is significantly associated with LVRR, independent of and superior to LV-GLS. Combined assessments of LV-MD and LV-GLS provide additive value beyond clinical variables for predicting LVRR in HFrEF.

BackgroundIndividuals with frailty are at increased risk of type 2 diabetes (T2D), but the underlying mechanisms are unclear. We aimed to investigate whether the frailty-T2D association is mediated by alterations in the metabolome and assess potential interaction with genetic susceptibility to diabetes.MethodsThis retrospective analysis, using data from a large prospective population-based cohort, included a total of 197,502 adults with baseline metabolomics data from the UK Biobank. Frailty was defined using the Fried frailty phenotype according to five components. Elastic net regression was applied to create a frailty-related metabolic signature. We assessed hazard ratios (HR) and its 95% confidence interval (CI) of incident T2D in relation to the baseline metabolic signature of frailty and examined the mediating role of the metabolic signature in the effect of frailty on T2D. Additive interaction between the metabolic signature and polygenic risk score for T2D (PRS-T2D) on the incidence of T2D was assessed as relative excess risk due to interaction (RERI).ResultsCompared with non-frailty, the HR (95% CI) of incident T2D in pre-frailty and frailty was 1.33 (1.26, 1.40) and 1.59 (1.46, 1.74), respectively. The metabolic signature of frailty (comprised of 53 metabolites) was positively associated with T2D risk (HR per standard deviation increment: 1.45; 95% CI: 1.42, 1.48), and explained 31.0% (95% CI: 25.8, 36.8) of the association between frailty and T2D. An additive interaction between metabolic signature of frailty and PRS-T2D was found (RERI: 9.43; 95% CI: 6.06, 12.80).ConclusionsThe increased risk of T2D in individuals with frailty may be mediated through effects on the metabolome, and the influence of such metabolic alterations on diabetes risk may be amplified in individuals with genetic susceptibility to T2D.

Pulmonary hypertension (PH) is a clinicopathological syndrome characterized by structural and functional alterations in the pulmonary vasculature arising from heterogeneous etiologies (including hypoxia) and diverse pathogenic mechanisms. These changes elevate pulmonary vascular resistance and increase pulmonary arterial pressure, ultimately progressing to right heart failure and potential fatality. Resistin-like molecule (RELM)-β activates multiple signaling pathways. This study aimed to explore the role of RELM-β in the development of chronic hypoxia-induced PH and its potential mechanisms. Exogenous human RELM-β was injected into a mouse model of hypoxia for 3 weeks, followed by histological and hemodynamic analyses. The relationship between RELM-β and membrane proteins or receptors (OR1N1, GIPC1 and CLIC4) was determined by affinity purification-mass spectrometry (AP-MS) and co-immunoprecipitation. At the same time, in vitro cell culture experiments were carried out. Cell membrane proteins or receptors (OR1N1, GIPC1, and CLIC4) were identified as proteins interacting with RELM-β and potentially involving in the development of PH. Compared with the RELM-β overexpression group, siRNA-mediated silencing of OR1N1, GIPC1, or CLIC4 resulted in significant reduction of cell viability in both human pulmonary artery smooth muscle cells (PASMCs) and human pulmonary arterial endothelial cells (PAECs). Moreover, augmenting effect of exogenous RELM-ß on the hypoxia-induced PH was remarkably reduced in the mice with genetic deficiency of GIPC1 (GIPC1 CKO) or CLIC4 (CLIC4 CKO) compared to the wild type mice. Findings of the current study suggested that RELM-β may play an important role in the development of hypoxia-induced PH through interacting with membrane proteins or receptors, including GIPC1, OR1N1, and CLIC4.

Previous studies have shown that ferroptosis of vascular smooth muscle cells (VSMCs) is involved in the development of aortic dissection (AD) and that histone methylation regulates this process. SP2509 acts as a specific inhibitor of lysine-specific demethylase 1 (LSD1), which governs a variety of biological processes. However, the effect of SP2509 on VSMC ferroptosis and AD remains to be elucidated. This aim of this study was to investigate the role and underlying mechanism of SP2509-mediated histone methylation on VSMC ferroptosis. Here, a mouse model of AD was established, and significantly reduced levels of H3K4me1 and H3K4me2 (target of SP2509) were found in the aortas of AD mice. In VSMCs, SP2509 treatment led to a dose-dependent increase in H3K4me2 levels. Furthermore, we found that SP2509 provided equivalent protection to ferrostatin-1 against VSMC ferroptosis, as evidenced by increased cell viability, decreased cell death and lipid peroxidation. RNA-sequencing analysis and subsequent experiments revealed that SP2509 counteracted cystine deficiency-induced response to inflammation and oxidative stress. More importantly, we demonstrated that SP2509 inhibited the expression of TFR and ferritin to reduce intracellular iron levels, thereby effectively blocking the process of ferroptosis. Therefore, our findings indicate that SP2509 protects VSMCs from multiple stimulus-induced ferroptosis by reducing intracellular iron levels, thereby preventing lipid peroxidation and cell death. These findings suggest that SP2509 may be a promising drug to alleviate AD by reducing iron deposition and VSMC ferroptosis.

Myocardial ischemia-reperfusion injury (MIRI) is a complex pathological process that results from the restoration of blood flow to ischemic myocardium, leading to a series of detrimental effects including oxidative stress and inflammation. Stachyose, a naturally occurring oligosaccharide found in traditional Chinese medicinal herbs, has been suggested to possess therapeutic properties against various pathological conditions. However, its impact on MIRI and the underlying mechanisms have not been fully elucidated. In this study, we aimed to investigate the therapeutic effects of stachyose on MIRI and to uncover the molecular mechanisms involved. Using both in vivo and in vitro models of MIRI, we evaluated the effects of stachyose on cardiac function and cell death pathways. Our results indicate that stachyose significantly improves cardiac function and reduces infarct size in MIRI mice. Mechanistically, stachyose modulates the ferroptotic pathway in cardiomyocytes by upregulating the expression of glutathione peroxidase 4 (GPX4) and reducing lipid peroxides and iron levels. Additionally, stachyose inhibits the pyroptotic pathway in macrophages by downregulating the expression of NLRP3, gasdermin D (GSMD-N), and cleaved-caspase-1, leading to decreased levels of proinflammatory cytokines interleukin (IL)-1 beta beta and IL-18. This study demonstrates that stachyose exerts a protective effect against MIRI by targeting both ferroptosis and pyroptosis pathways, suggesting its potential as a novel therapeutic agent for the treatment of MIRI. Further research is warranted to explore the detailed mechanisms and therapeutic potential of stachyose in clinical settings.

Tissue engineering heart valves (TEHVs) are expected to address the limitations of mechanical and bioprosthetic valves used in clinical practice. Decellularized heart valve (DHV) is an important scaffold of TEHVs due to its natural three-dimensional structure and bioactive extracellular matrix, but its mechanical properties and hemocompatibility are impaired. In this study, DHV is cross-linked with three different molecular weights of oxidized hyaluronic acid (OHA) by a Schiff base reaction and presented enhanced stability and hemocompatibility, which could be mediated by the molecular weight of OHA. Notably, DHV cross-linked with middle- and high-molecular-weight OHA could drive the macrophage polarization toward the M2 phenotype in vitro. Moreover, DHV cross-linked with middle-molecular-weight OHA scaffolds are further modified with RGD-PHSRN peptide (RPF-OHA/DHV) to block the residual aldehyde groups of the unreacted OHA. The results show that RPF-OHA/DHV not only exhibits anti-calcification properties, but also facilitates endothelial cell adhesion and proliferation in vitro. Furthermore, RPF-OHA/DHV shows excellent performance under an in vivo hemodynamic environment with favorable recellularization and immune regulation without calcification. The optimistic results demonstrate that OHA with different molecular weights has different cross-linking effects on DHV and that RPF-OHA/DHV scaffold with enhanced immune regulation, anti-calcification, and recellularization properties for clinical transformation.

Purpose:Inflammation is a hallmark of the initial development and progression of aortic dissection. This study aimed to investigate the predictive value of preoperative neutrophils in aorta-related adverse events (AAEs) after thoracic endovascular aortic repair (TEVAR) for type B aortic dissection (TBAD).Patients and Methods:A total of 80 patients with TBAD undergoing TEVAR were enrolled in our hospital. Preoperative inflammatory markers, including white blood cells (WBCs), neutrophils, neutrophil-to-lymphocyte ratio (NLR) and plasma high-sensitivity C-reactive protein (hs-CRP), were measured. Circulating neutrophil subpopulation was determined by flow cytometry. Kaplan-Meier curve was performed to determine whether neutrophil subsets independently predicted aorta-related adverse events (AAEs) after TEVAR.Results:Compared with control group, the prevalence of hypertension and the levels of inflammatory indicators including WBCs, total neutrophils, NLR, immature neutrophils and hs-CRP were significantly higher in TBAD patients. Receiver operating characteristic (ROC) curve showed that NLR, absolute number of total neutrophils and percent CD10- immature neutrophils had excellent area under curves. During the 18-month follow-up, 16 (20.0%) were reported to occur AAEs, while 4 deaths (5.0%) were documented. Percent immature neutrophil was markedly higher in TBAD patients experiencing AAEs as compared with those without AAEs. Kaplan-Meier curve and Cox regression analysis demonstrated that percent immature neutrophil was the only predictor correlated with the occurrence of AAEs (hazard ratio 7.66, 95% CI: 2.91, 20.17, P = 0.018).Conclusion:Increased CD10- immature neutrophils could act as a potential biomarker related to long-term adverse outcomes in TBAD patients following TEVAR.

Introduction: Dilated cardiomyopathy (DCM) represents a diverse set of myocardial diseases characterized by notable genetic heterogeneity. Although over 50 genes have been associated with DCM, these collectively explain 35% of idiopathic DCM cases. Variants in the FBN1 gene encoding fibrillin-1 are primarily linked to connective tissue disorders. Considering the potential of these disorders to impact myocardial tissue, this study probes into the possible association between FBN1 variants and DCM. Aim: The objective of this study was to investigate the association between FBN1 variants and DCM in a Chinese Han population. Methods and Results: We performed whole-exome sequencing (WES) to identify rare FBN1 variants among 1,059 DCM cases and 514 controls. Utilizing a case-control strategy and the optimal sequence kernel association test (SKAT-O), we found a significant enrichment of rare deleterious FBN1 variants in DCM patients (19 of 1,059 vs. 0 of 514, PSKAT-O = 7.49E-04). Clinical characteristics analysis indicated a higher occurrence of atrial fibrillation and a higher rate of implantable cardioverter-defibrillator (ICD) implantation among DCM patients carrying FBN1 variants (FBN1+) compared to non-carriers (FBN1-). However, these FBN1 variants did not significantly affect primary endpoints, defined as cardiac mortality or heart transplantation, yet appeared to increase the risk of secondary endpoints, including all-cause mortality or heart failure recurrence. Conclusion: The findings suggest an association between rare deleterious variants in the FBN1 gene and DCM in a Chinese Han population. Our findings underline the importance of further research to validate these results and elucidate the role of FBN1 in DCM. Potential Impact of the findings: This research provides fresh insights into the potential role of FBN1 rare variants in DCM, pointing to new directions for future genetic studies and potential therapeutic strategies in DCM management.

Background: Abdominal aortic aneurysms (AAA) are chronic inflammation in nature and are closely related to macrophages. The purpose was to explore regulating macrophage polarization with target-macrophage nanoparticles impacting the development of AAA.Methods: Galactose-modified nanoparticles were prepared by self-assembly technology for delivering microRNA (miR)-223. In AngiotensinII-induced experimental AAA model, miR-223-loaded nanoparticles (MirNPs) or PBS was injected at day 7 before and after operation, respectively. Cultured cells and aortic specimen were collected to be analyzed with histology and biochemical examination.Results: In vitro, miR-223 promoted bone marrow-derived macrophages (BMDMs) to polarize to M2. In experimental AAA model, MirNPs significantly decreased the AAA incidence and the ratio of M1 macrophages and production of related proinflammatory cytokines. Furthermore, MirNPs also reduced the expression of the NLRP3 inflammasome.Conclusion: Our findings suggested that miR-223-loaded nanoparticles targeting macrophage polarization may mitigate AAA progression via downregulating of NLRP3.(c) 2022 Published by Elsevier Inc.

BackgroundObservational studies have revealed that dried fruit intake may be associated with cancer incidence; however, confounding factors make the results prone to be disturbed. Therefore, we conducted a two-sample Mendelian randomization (MR) study to explore the causal relationship between dried fruit intake and 11 site-specific cancers.Materials and methodsForty-three single nucleoside polymers (SNPs) with robust genome-wide association study (GWAS) evidence, strongly correlated with dried fruit intake, were used as instrumental variables (IVs) in this study. The summary-level genetic datasets of site-specific cancers were obtained from the Oncoarray oral cavity and oropharyngeal cancer consortium, International Lung Cancer Consortium, Breast Cancer Association Consortium (BCAC), Ovarian Cancer Association Consortium, PanScan1, and GWAS of other scholars. We analyzed the causality between dried fruit intake and 11 site-specific cancers using the inverse-variance-weighted (IVW) and weighted median (WM) methods. For the results of the MR analysis, Cochran’s Q test was used to check for heterogeneity, and multiplicative random effects were used to evaluate the heterogeneity further. Gene pleiotropy was tested using MR-Egger regression and MR-PRESSO methods. In addition, the main results of this study were validated by using the summary statistical data from the FinnGen and UK Biobank databases, and adjusted body mass index (BMI), years of education, fresh fruit intake, and vitamin C using multivariable MR analysis to ensure the stability of the research results.ResultsThe evidence from IVW analyses showed that each increase of dried fruit intake by one standard deviation was statistically significantly associated with 82.68% decrease of oral cavity/pharyngeal cancer incidence risk (P = 0.0131), 67.01% decrease of lung cancer incidence risk (P = 0.0011), 77% decrease of squamous cell lung cancer incidence risk (P = 0.0026), 53.07% decrease of breast cancer incidence risk (P = 4.62 × 10–5), 39.72% decrease of ovarian cancer incidence risk (P = 0.0183), 97.26% decrease of pancreatic cancer incidence risk (P = 0.0280), 0.53% decrease of cervical cancer incidence risk (P = 0.0482); however, there was no significant effect on lung adenocarcinoma (P = 0.4343), endometrial cancer (P = 0.8742), thyroid cancer (P = 0.6352), prostate cancer (P = 0.5354), bladder cancer (P = 0.8996), and brain cancer (P = 0.8164). In the validation part of the study results, the causal relationship between dried fruit intake and lung cancer (P = 0.0043), squamous cell lung cancer (P = 0.0136), and breast cancer (P = 0.0192) was determined. After adjusting for the potential impact of confounders, the causal relationship between dried fruit intake and lung cancer (P = 0.0034), squamous cell lung cancer (P = 0.046), and breast cancer (P = 0.0001) remained. The sensitivity analysis showed that our results were stable and reliable.ConclusionThe intake of dried fruits may have a protective effect against some site-specific cancers. Therefore, health education and a reasonable adjustment of dietary proportions may help in the primary prevention of cancer.

Background:The misdiagnosis of aortic dissection (AD) can lead to a catastrophic prognosis. There is currently a lack of stable serological indicators with excellent efficacy for the differential diagnosis of AD and coronary artery disease (CAD). A recent study has shown an association between AD and iron metabolism. Thus, we investigated whether iron metabolism could discriminate AD from CAD.Methods:This retrospective and multicenter cross-sectional study investigated the efficacy of biomarkers of iron metabolism for the differential diagnosis of AD. We collected biomarkers of iron metabolism, liver function, kidney function, and other biochemistry test, and further, logistic regression analysis was applied.Results:Between Oct. 8, 2020, and Mar. 1, 2021, we recruited 521 patients diagnosed with AD, CAD, and other cardiovascular diseases (OCDs) with the main symptoms of chest and back pain and assigned them to discovery set (n = 330) or validation set (n = 191). We found that six serum biomarkers, including serum iron, low-density lipoprotein, uric acid, transferrin, high-density lipoprotein, and estimated glomerular filtration rate, can serve as a novel comprehensive indicator (named FLUTHE) for the differential diagnosis of AD and CAD with a sensitivity of 0.954 and specificity of 0.905 to differentially diagnose AD and CAD more than 72 h past symptom onset.Conclusion:Our findings provide insight into the role of iron metabolism in diagnosing and distinguishing AD, which might in the future be a key component in AD diagnosis. Furthermore, we establish a novel model named "FLUTHE" with higher efficiency, safety, and economy, especially for patients with chest pain for more than 72 h.

A variety of programmed cell death types have been shown to participate in the loss of smooth muscle cells (SMCs) during the development of aortic dissection (AD), but it is still largely unclear whether ferroptosis is involved in the development of AD. In the present study, we found that the expression of key ferroptosis regulatory proteins, solute carrier family 7 member 11 (SLC7A11), ferroptosis suppressor protein 1 (FSP1) and glutathione peroxidase 4 (GPX4) were downregulated in aortas of Stanford type A AD (TAAD) patients, and liproxstatin-1, a specific inhibitor of ferroptosis, obviously abolished the β-aminopropionitrile (BAPN)-induced development and rupture of AD in mice. Furthermore, the expression of methyltransferase-like 3 (METTL3), a major methyltransferase of RNA m6A, was remarkably upregulated in the aortas of TAAD patients, and the protein levels of METTL3 were negatively correlated with SLC7A11 and FSP1 levels in human aortas. Overexpression of METTL3 in human aortic SMCs (HASMCs) inhibited, while METTL3 knockdown promoted SLC7A11 and FSP1 expression. More importantly, overexpression of METTL3 facilitated imidazole ketone erastin- and cystine deprivation-induced ferroptosis, while knockdown of METTL3 repressed ferroptosis of HASMCs. Overexpression of either SLC7A11 or FSP1 largely abrogated the effect of METTL3 on HASMC ferroptosis. Therefore, we have revealed that ferroptosis is a critical cause of AD in both humans and mice and that METTL3 promotes ferroptosis of HASMCs by inhibiting the expression of SLC7A11 and FSP1. Thus, targeting ferroptosis or m6A RNA methylation is a potential novel strategy for the treatment of AD.

Abdominal aortic aneurysm (AAA) is characterized by abdominal aorta dilatation and progressive structural impairment and is usually an asymptomatic and potentially lethal disease with a risk of rupture. To investigate the underlying mechanisms of AAA initiation and progression, seven AAA datasets related to human and mice were downloaded from the GEO database and reanalysed in the present study. After comprehensive bioinformatics analysis, we identified the enriched pathways associated with inflammation responses, vascular smooth muscle cell (VSMC) phenotype switching and cytokine secretion in AAA. Most importantly, we identified ATPase Na+/K+ transporting subunit alpha 2 (ATP1A2) as a key gene that was significantly decreased in AAA samples of both human and mice; meanwhile, its reduction mainly occurred in VSMCs of the aorta; this finding was validated by immunostaining and Western blot in human and mouse AAA samples. Furthermore, we explored the potential upstream transcription factors (TFs) that regulate ATP1A2 expression. We found that the TF AT-rich interaction domain 3A (ARID3A) bound the promoter of ATP1A2 to suppress its expression. Our present study identified the ARID3A-ATP1A2 axis as a novel pathway in the pathological processes of AAA, further elucidating the molecular mechanism of AAA and providing potential therapeutic targets for AAA.

Background Vascular smooth muscle cell (VSMC) phenotype switching is critical for neointima formation, which is the major cause of restenosis after stenting or coronary artery bypass grafting. However, the epigenetic mechanisms regulating phenotype switching of VSMCs, especially histone methylation, are not well understood. As a main component of histone lysine demethylases, Jumonji demethylases might be involved in VSMC phenotype switching and neointima formation. Methods and results A mouse carotid injury model and VSMC proliferation model were constructed to investigate the relationship between histone methylation of H3K36 (downstream target molecule of Jumonji demethylase) and neointima formation. We found that the methylation levels of H3K36 negatively correlated with VSMC proliferation and neointima formation. Next, we revealed that JIB-04 (a pan-inhibitor of the Jumonji demethylase superfamily) could increase the methylation levels of H3K36. Furthermore, we found that JIB-04 obviously inhibited HASMC proliferation, and a cell cycle assay showed that JIB-04 caused G2/M phase arrest in HASMCs by inhibiting the phosphorylation of RB and CDC2 and promoting the phosphorylation of CHK1. Moreover, JIB-04 inhibited the expression of MMP2 to suppress the migration of HASMCs and repressed the expression of contraction-related genes. RNA sequencing analysis showed that the biological processes associated with the cell cycle and autophagy were enriched by using Gene Ontology analysis after HASMCs were treated with JIB-04. Furthermore, we demonstrated that JIB-04 impairs autophagic flux by downregulating STX17 and RAB7 expression to inhibit the fusion of autophagosomes and lysosomes. Conclusion JIB-04 suppresses the proliferation, migration, and contractile phenotype of HASMCs by inhibiting autophagic flux, which indicates that JIB-04 is a promising reagent for the treatment of neointima formation.

Background: The misdiagnosis of aortic dissection (AD) can lead to a catastrophic prognosis. There is currently a lack of stable serological indicators with excellent efficacy for the differential diagnosis of AD and coronary artery disease (CAD). Recent study has shown association between AD and iron metabolism. Thus, we investigated whether iron metabolism could discriminate AD from CAD.Methods: In this retrospective and multicenter cross-sectional study that investigated the efficacy of biomarkers of iron metabolism for the differential diagnosis of AD. We collected biomarkers of iron metabolism, liver function, kidney function and other biochemistry test and further Logistic regression analysis was applied. Findings: Between Oct. 8, 2020, to Mar. 1, 2021, we recruited 521 patients diagnosed with AD, CAD, and other cardiovascular diseases (OCDs) with the main symptoms of chest and back pain and assigned them to discovery set (n = 330) or validation set (n = 191). We found that six serum biomarkers, serum iron, low-density lipoprotein, uric acid, transferrin, high-density lipoprotein and estimated glomerular filtration rate, can serve as a novel comprehensive indicator (named FLUTHE) for the differential diagnosis of AD and CAD with sensitivity up to 95.4% and specificity 90.5% to differentially diagnose AD and CAD more than 72 hours past symptom onset.Interpretation: Iron metabolism showed great relationship with aortic dissection. Results suggest the novel combination, FLUTHE, was an ideal indicator in diagnosing AD especially when time from symptom onset was over 72 hours. Funding Information: The National Natural Science Foundation of China (NO. 82070488 and NO. 82000440).Declaration of Interests: The authors declare that they have no competing interests.Ethics Approval Statement: The study was approved by the Ethics Review Board of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology.