OBJECTIVES:Insulin-like growth factor binding protein-4 (IGFBP-4) fragments generated by PAPP-A-mediated proteolysis are emerging biomarkers of myocardial stress and remodeling. Although elevated N-terminal IGFBP-4 (NT-IGFBP-4) has been associated with heart failure (HF), its prognostic value remains unclear. This study evaluated the prognostic performance of NT-IGFBP-4, measured using a validated chemiluminescent immunoassay, in patients with acute heart failure (AHF) and compared it with the conventional biomarker NT-proBNP. METHODS:A total of 309 patients with AHF were included. Baseline NT-IGFBP-4 and NT-proBNP were measured, and patients were followed for up to 1 year for HF rehospitalization and all-cause mortality. The prognostic performance of each biomarker and their combination for 30-day and 1-year outcomes was evaluated using ROC analysis, Kaplan-Meier analysis, and Cox proportional hazards regression with multivariable adjustment for clinical factors and the other biomarker. Optimal cut-off values were derived from ROC analysis. RESULTS:During follow-up, 81 patients experienced clinical events, including 10 deaths and 71 hospital readmissions. NT-IGFBP-4 showed a moderate correlation with NT-proBNP, but only weak associations with hs-CRP and hs-cTnI. For 30-day outcomes, NT-IGFBP-4 remained independently predictive after mutual adjustment (HR = 10.07[3.10-32.68], p < 0.001), whereas NT-proBNP lost statistical significance under the same conditions (HR = 2.68[0.72-10.01], p = 0.142). For 1-year outcomes, both NT-IGFBP-4 (HR = 3.27[1.80-5.91], p < 0.001) and NT-proBNP (HR = 2.04[1.18-3.53], p = 0.010) remained independent predictors after adjustment for clinical covariates and each other. Notably, the combined assessment of both biomarkers further enhanced 1-year risk discrimination (HR = 5.72[2.73-11.98], p < 0.001). CONCLUSIONS:NT-IGFBP-4 is a novel, independent prognostic biomarker in AHF, supporting its use in multimarker strategies for patient management.

Objective:This study aimed to assess the clinical utility and prognostic value of a panel of serum cytokines for predicting major adverse cardiovascular events (MACE) in patients with acute myocardial infarction (AMI) after percutaneous coronary intervention (PCI). Methods:We enrolled 232 AMI patients who underwent PCI. Based on the occurrence of MACE during follow-up, patients were categorized into a poor prognosis group (n=127) and a good prognosis group (n=105). Serum levels of interleukin IL-1β, IL-6, IL-8, IL-10, and tumor necrosis factor-alpha (TNF-α) were measured. Spearman correlation analysis was employed to evaluate the relationships between cytokine levels and MACE risk. Nine distinct machine learning models, incorporating all identified independent risk factors for MACE, were developed and evaluated. Results:Levels of high-sensitivity C-reactive protein (hs-CRP), IL-6, IL-8, and TNF-α were significantly elevated in patients who developed MACE compared to those who did not. IL-6 emerged as a relatively strong predictor of MACE, with an area under the curve (AUC) of 0.82 (95% CI: 0.77-0.87). The IL-6/IL-10 ratio also demonstrated predictive value for MACE (AUC=0.74, 95% CI: 0.69-0.80). Furthermore, significant differences were observed between initial presentation and readmission levels for the IL-8/IL-10 and TNF-α/IL-10 ratios in patients readmitted for AMI following MACE. Among the nine machine learning models constructed, the XGBoost and Logistic Regression models, which incorporated all independent risk factors, demonstrated robust performance, both AUC values exceeded 0.8. Conclusion:AMI patients with elevated serum cytokine levels exhibited a lower 1-year survival rate. Serum cytokine profiles show significant predictive value for MACE following PCI in AMI patients.

Objective:The etiology and specific pathological mechanisms of keloids remain elusive. Array expression profiling has revealed dysregulation of the transcription cofactor ankyrin repeat domain 1 (ANKRD1) in keloid fibroblasts. The present study focused on examining the expression pattern of ANKRD1 in keloids and assessing its function in human keloid fibroblasts (HKFs). Material and Methods:Differential mRNA expression profiles in keloid fibroblasts were investigated by analyzing data from gene expression omnibus (GEO) datasets. Immunohistochemistry assays were performed to verify the expression patterns of ANKRD1 and claudin 11 (CLDN11) in keloid tissue samples. Functional studies were conducted by transfecting HKFs with either a small interfering RNA (siRNA) targeting ANKRD1 (siANKRD1) or ANKRD1-overexpressing plasmids. The functional impact of ANKRD1 was assessed using cell proliferation, flow cytometry, and Transwell migration assays. mRNA expression was evaluated using reverse transcription polymerase chain reaction, and protein expression was determined using Western blotting. Results:Analysis of the GEO series (GSE) GSE44270 revealed eight differentially expressed mRNAs, with ANKRD1 and CLDN11 being the top two downregulated mRNAs. ANKRD1 expression was observed to be lower in keloid tissues than in normal skin tissues, whereas CLDN11 expression showed no significant difference between the two groups. ANKRD1 overexpression suppressed HKF proliferation, migration, and the expression levels of collagen I, fibronectin, matrix metallopeptidase 9, whereas the opposite effects were observed on ANKRD1 knockdown. ANKRD1 did not affect apoptotic cell levels. Conclusion:ANKRD1 is downregulated in keloids and inhibits the growth, migration, and extracellular matrix deposition of keloid fibroblasts. Thus, ANKRD1 may function as a suppressor in keloid formation.

OBJECTIVE:Autoimmune diseases, such as systemic lupus erythematosus (SLE), are associated with pulmonary arterial hypertension (PAH), a condition that can lead to heart failure. However, whether T cells also contribute to the occurrence of PAH in SLE has not been clarified. The objective of this study was to elucidate the role of Activin A and activated receptor signaling in SLE-PAH. METHODS:Mass Cytometry (CyTOF) analysis was performed to identify the major affected immune cell population in patients with SLE-PAH. Serum Activin A and interleukin-17 (IL-17) levels in patients with SLE-PAH, patients with SLE, and healthy donors were determined by enzyme-linked immunosorbent assay. Cocultures of Th17 cells with pulmonary microvascular endothelial cells (PMECs) and relevant rodent models were used to identify the converged target. RESULTS:The reduced CD4+ T cell number was detected in patients with SLE-PAH after treatment with immunosuppressant and vasodilator. Increased Th17 cells population and higher serum Activin A and IL-17 levels were found in patients with SLE-PAH compared to patients with SLE only or donors. Activin A signals via activin receptor-like kinase 4 (ALK4) in both Th17 cells and PMECs. Overexpressing ALK4 in Th17 cells increased IL-6 and endothelial-mesenchymal transition (EndoMT) marker levels in cocultured PMECs. We found severe SLE-pulmonary hypertension (PH) in mice by overexpressing ALK4, and alleviated hemodynamic changes in CD4+ T cells depletion mice. ALK4 inhibitor vactosertib (TEW-7197) effectively treated SLE-PH mice by repressing connective tissue growth factor (CTGF) transcription, which was induced by ALK4 activated pSmad2 and pSTAT3. CONCLUSION:Our findings suggest that Activin A activates ALK4 in Th17 cells, thereby inducing IL-17 secretion. Concurrently, activated ALK4 induces EndoMT in human PMECs (hPMECs) via CTGF up-regulation. It suggests that ALK4 is a promising therapeutic target for SLE-PAH.

OBJECTIVES:Insulin-like growth factor binding protein-4 (IGFBP-4) fragments are reported as emerging biomarkers for cardiovascular disease (CVD) risk assessment. To ensure data reliability and improve clinical application, the first automatic chemiluminescent immunoassay (CLIA) for NT-IGFBP-4 was developed and its distribution across CVDs was evaluated in this study. METHODS:Fragment-specific monoclonal antibodies were used to develop immunoassay, followed by comprehensive analytical validation, including limit of blank (LoB), limit of detection (LoD), limit of quantification (LoQ), linearity, specificity, precision, sample type compatibility, and stability. Reference intervals for NT-IGFBP-4 were established in healthy individuals, with variations analyzed based on gender, age, body mass index (BMI), and renal function. Additionally, NT-IGFBP-4 distribution was explored in CVD patients. RESULTS:The newly developed chemiluminescence assay demonstrated high specificity for NT-IGFBP-4, with excellent sensitivity (LoQ=1.0 ng/mL), broad linearity (1.0-1,000.0 ng/mL), and strong precision (CV≤3.0 %). It showed no interference from common endogenous substances, maintained compatible with various sample types, and remained stable under different storage conditions. Reference intervals showed slight variations by gender and age, with levels being independent of BMI but influenced by renal function. NT-IGFBP-4 levels were significantly elevated in CVDs, especially in heart failure, correlating with NYHA classification and LVEF (%), with higher levels indicating worse cardiac function. CONCLUSIONS:The new automatic NT-IGFBP-4 (CLIA) assay offers a highly specific, sensitive and precise method for quantifying IGFBP-4 fragments. Its validated performance and disease association findings would enhance its diagnostic and prognostic potential in CVD research, particularly in heart failure.

Pulmonary veno-occlusive disease (PVOD) is a fatal disease characterized by the remodelling of pulmonary veins and haemosiderin accumulation in macrophages. Although (General Control Nonderepressible 2) GCN2 deficiency has been reported in PVOD patients, the underlying mechanism by which GCN2 deficiency affects the pulmonary venous cells and the surrounding cells, remains unclear. Here, we perform immunohistochemistry and scRNA-sequencing analyses to show that macrophages are the major population affected by GCN2 deficiency and ferroptosis pathway-related genes are upregulated in lung macrophages of PVOD patients. Treatment with the specific ferroptosis inhibitor ferrostatin-1 (Fer-1) reverses the changes in haemodynamic indices observed in Eif2ak4K1488X/K1488X hypoxia mice and PVOD model rats. Furthermore, GCN2 deficiency increases HMOX1 and iron levels to facilitate ferroptosis in macrophages, and enhances arterial marker expression in venous endothelial cells (VECs). Specifically, spatial transcriptome analysis shows increased expression of NRP1, KDR and EFNB2 through ETS1 in VECs from PVOD patients. Our findings suggest the potential of targeting macrophage ferroptosis as a therapeutic strategy for treating related vascular diseases, and of using NRP1/KDR/EFNB2 expression as a specific marker set for venous arterialization.

OBJECTIVES:The potential relationship between cardiac troponin (cTn) composition and etiologies of myocardial injury may provide diagnostic insights for myocardial infarction (MI). We compared the performance of novel ternary cTnI-cTnT-TnC complex (ITC) assays with high-sensitivity (hs)-cTnI and hs-cTnT in differentiating MI patients at presentation. METHODS:Plasma samples from 1,210 patients with suspected MI were collected. Major cTn forms were quantified using long-cTnT ITC complex, hs-total ITC complex, hs-cTnI and hs-cTnT assay. We compared the assay performance based on the 99th percentile upper reference limits (URLs), evaluated their area under the receiver operating characteristic curve (AUC), and assessed their ability to differentiate MI using established cut-off values. RESULTS:A total of 138 (11.4 %) patients were diagnosed with MI. 86 % patients had long-cTnT ITC complex concentration below the 99th percentile URLs (sensitivity: 0.818, specificity: 0.943, NPV: 0.975, PPV: 0.649). AUC was 0.959 for long-cTnT ITC complex, comparable to hs-cTnI (0.953) and superior to hs-total ITC complex (0.889) and hs-cTnT (0.927). Using established cut-off values, the long-cTnT ITC complex assay classified 10 % of patients as rule-in and 52 % as rule-out (sensitivity: 0.993, NPV: 0.998, specificity: 0.972, PPV: 0.760), which was comparable to hs-cTnI and better than hs-total ITC complex and hs-cTnT. In early-presenting patients, the long-cTnT ITC complex demonstrated enhanced discrimination and improved diagnostic performance. CONCLUSIONS:The long-cTnT ITC complex assay demonstrated performance comparable to hs-cTnI and superior to hs-total ITC complex and hs-cTnT in distinguishing MI using a single sample at presentation. These findings suggest its potential utility in improving risk stratification and triage strategies, particularly among early-presenting patients.

Objective:This study aims to identify and validate key genes involved in the progression of myocardial infarction (MI) and to investigate their roles in inflammatory response, immune regulation, and myocardial remodeling. A systematic analysis will be conducted using bioinformatics and machine learning methods. Methods:Gene expression data of GSE60993, GSE61144, GSE66360 and GSE48060 from four datasets were collected from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) between MI samples and normal samples were screened by the limma package. Weighted gene co-expression network analysis (WGCNA) was employed to identify genetic modules associated with MI. Core genes in key modules were screened using LASSO regression and support vector machine recursive feature elimination (SVM-RFE). These genes were then subjected to functional enrichment analysis, including Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO), and Gene Set Enrichment Analysis (GSEA). The CIBERSORT algorithm was utilized to evaluate immune cell infiltration patterns. Finally, potential therapeutic targets were explored through drug-gene interaction analysis using the DGIdb database. Results:After correcting for batch effects across datasets, we identified 687 differentially expressed genes (DEGs), including 405 upregulated and 282 downregulated genes. WGCNA analysis identified a highly correlated module with MI (turquoise module) containing 324 genes. Integrative machine learning (LASSO regression and SVM-RFE) and validation identified five key MI-associated genes: ANPEP, S100A9, MMP9, DAPK2, and FCAR. These genes were functionally enriched in inflammatory and immune-related pathways and correlated with immune cell infiltration, particularly neutrophils and macrophages. Notably, S100A9, FCAR, and MMP9 emerged as druggable targets. Conclusion:The five hub genes identified in this study (ANPEP, S100A9, MMP9, DAPK2, and FCAR) significantly contribute to MI development by modulating inflammatory responses and immune regulation. Their strong association with MI pathogenesis highlights their potential as diagnostic markers and therapeutic targets, which may lead to new clinical applications for MI management.

Objectives Cardiac troponin (cTn) is the key biomarker for diagnosis of acute coronary syndrome (ACS). We performed a complete assessment of the high-sensitivity cardiac troponin I (hs-cTnI) (CLIA) assay on the analytical performance and clinical diagnostic performance, which was compared with Abbott ARCHITECT hs-cTnI assay. Methods Sex-specific 99th percentile upper reference limits (URLs) were determined from a healthy population of 424 males and 408 females. High-sensitivity performance was assessed by examining the imprecision at sex-specific URLs and the detectable results above LoD in a cohort of healthy population. The diagnostic performance of the hs-cTnI (CLIA) assay was validated in a population of 934 patients with suspected ACS. Results The 99th percentile URLs were 15.3 ng/L for female, 31.3 ng/L for male and 24.2 ng/L for overall population. The total imprecision near the sex-specific 99th percentile URLs were <5 %. 76.74 % of females, 97.12 % of males and 86.69 % of overall population had cTnI values exceeding the LoD, which met the criteria of high-sensitivity troponin assay. No cross-reactivity or interference was identified. The diagnostic sensitivity, specificity, PPV, NPV, and AUC of hs-cTnI (CLIA) assay were 97.97 , 90.70, 79.02, 99.21 % and 0.9885, respectively, which were comparable to ARCHITECT hs-cTnI assay. Conclusions hs-cTnI (CLIA) assay is a high-sensitivity troponin I method with high precision, sensitivity and specificity. The clinical diagnostic performance of hs-cTnI (CLIA) is comparable to the established ARCHITECT hs-cTnI assay. Mindray’s hs-cTnI (CLIA) assay is an attractive alternative for diagnosis of myocardial infarction with a high level of accuracy and safety.

BACKGROUND:Current studies suggest that cardiac troponin (cTn) forms in the circulation may vary in different clinical scenarios. Our aim was to design a combination of cTn assays specific to the main cTn forms and to evaluate their analytical performance. METHODS:We developed immunoassays specific for measuring (1) long-cTnT cTnI-cTnT-TnC (ITC) ternary complex, with cTnT in long form without cleavage at the C-terminal amino acids residue 189-223, designated "long-cTnT ITC complex assay;" (2) both the long-cTnT ITC complex plus short-cTnT ITC complex, designated "hs-total ITC complex assay;" (3) the central part of cTnT of both the long-cTnT ITC complex and free cTnT, designated "hs-cTnT assay." Sex-specific 99th percentile upper reference limits (URLs) were determined. High-sensitivity performance was assessed by examining the imprecision and detectable results above limit of detection (LoD) in the healthy population. RESULTS:Both complex immunoassays exhibited excellent analytical sensitivity, precision, and specificity. The 99th percentile URLs were as follows: long-cTnT ITC complex: male 0.90 ng/L, female 0.87 ng/L; hs-total ITC complex: male 16.15 ng/L, female 10.08 ng/L; hs-cTnT: male 15.57 ng/L, female 14.28 ng/L. The total imprecision at or below the sex-specific 99th percentile URLs was <5% for all assays. The hs-total ITC complex and the hs-cTnT assays showed >50% of measurable concentrations above the LoD. However, <20% were measurable for the long-cTnT ITC complex assay. CONCLUSIONS:The cTn assays detected concentrations of major cTn forms in the circulation with high sensitivity, precision, and specificity, supporting their use for monitoring cTn complex and fragmentation forms during myocardial injuries.

BACKGROUND:Increased cardiac troponin (cTn) concentrations occur in acute myocardial injury and chronic diseases. Characterization of cTn composition in the circulation may assist in differentiating etiologies of myocardial injury. Our goal was to study cTn composition and kinetics in patients following type 1 myocardial infraction (T1MI), cardiac procedures, and chronic heart diseases to establish the relationship between cTn composition and clinical diagnosis. METHODS:Plasma samples were collected from 201 patients with T1MI, 78 undergoing cardiac surgeries, and 218 with chronic cardiomyopathy or chronic heart failure. Major cTn forms in the circulation and their ratios were analyzed using cTn composition immunoassays, targeting (a) the long-cTnT cTnI-cTnT-TnC (ITC) ternary complex, short-cTnT ITC complex cleaved at amino acids residues 189-223 of cTnT, and the binary cTnI-TnC (IC) complex, and designated the "high-sensitivity (hs)-cTnI assay;" (b) the long-cTnT ITC complex, and designated the "long-cTnT ITC complex assay;" (c) the long-cTnT ITC complex and short-cTnT ITC complex, and designated the "hs-total ITC complex assay;" and (d) the central part of cTnT of both the long-cTnT ITC complex and free cTnT, and designated the "hs-cTnT assay." RESULTS:Early-stage T1MI patients showed a high ratio of long-cTnT ITC complex to cTnI (long-cTnT ITC complex/cTnI, R1). Similarly, patients after acute cardiac surgery exhibited increased cTn concentrations with high R1, which decreased rapidly. In chronic disease, cTn composition exhibited stable and low R1 and high ratios of cTnT to cTnI (cTnT/cTnI, R3). CONCLUSIONS:Kinetic differences in multiple cTn forms contribute to the differentiation between acute injury and chronic disease, with a high proportion of long-cTnT ITC complex implying occurrence of acute injury.

The accumulation and spread of prion-like proteins is a key feature of neurodegenerative diseases (NDs) such as Alzheimer's disease, Parkinson's disease, or Amyotrophic Lateral Sclerosis. In a process known as 'seeding', prion-like proteins such as amyloid beta, microtubule-associated protein tau, α-synuclein, silence superoxide dismutase 1, or transactive response DNA-binding protein 43 kDa, propagate their misfolded conformations by transforming their respective soluble monomers into fibrils. Cellular and molecular evidence of prion-like propagation in NDs, the clinical relevance of their 'seeding' capacities, and their levels of contribution towards disease progression have been intensively studied over recent years. This review unpacks the cyclic prion-like propagation in cells including factors of aggregate internalization, endo-lysosomal leaking, aggregate degradation, and secretion. Debates on the importance of the role of prion-like protein aggregates in NDs, whether causal or consequent, are also discussed. Applications lead to a greater understanding of ND pathogenesis and increased potential for therapeutic strategies.

Background Autism Spectrum Disorders (ASD) are defined as a group of pervasive neurodevelopmental disorders, and the heterogeneity in the symptomology and etiology of ASD has long been recognized. Altered immune function and gut microbiota have been found in ASD populations. Immune dysfunction has been hypothesized to involve in the pathophysiology of a subtype of ASD. Methods A cohort of 105 ASD children were recruited and grouped based on IFN-γ levels derived from ex vivo stimulated γδT cells. Fecal samples were collected and analyzed with a metagenomic approach. Comparison of autistic symptoms and gut microbiota composition was made between subgroups. Enriched KEGG orthologues markers and pathogen-host interactions based on metagenome were also analyzed to reveal the differences in functional features. Results The autistic behavioral symptoms were more severe for children in the IFN-γ-high group, especially in the body and object use, social and self-help, and expressive language performance domains. LEfSe analysis of gut microbiota revealed an overrepresentation of Selenomonadales, Negatiyicutes, Veillonellaceae and Verrucomicrobiaceae and underrepresentation of Bacteroides xylanisolvens and Bifidobacterium longum in children with higher IFN-γ level. Decreased metabolism function of carbohydrate, amino acid and lipid in gut microbiota were found in the IFN-γ-high group. Additional functional profiles analyses revealed significant differences in the abundances of genes encoding carbohydrate-active enzymes between the two groups. And enriched phenotypes related to infection and gastroenteritis and underrepresentation of one gut–brain module associated with histamine degradation were also found in the IFN-γ-High group. Results of multivariate analyses revealed relatively good separation between the two groups. Conclusions Levels of IFN-γ derived from γδT cell could serve as one of the potential candidate biomarkers to subtype ASD individuals to reduce the heterogeneity associated with ASD and produce subgroups which are more likely to share a more similar phenotype and etiology. A better understanding of the associations among immune function, gut microbiota composition and metabolism abnormalities in ASD would facilitate the development of individualized biomedical treatment for this complex neurodevelopmental disorder.

BACKGROUND: Endothelium dysfunction is a central problem for early rejection due to the host alloimmune response and the late status of arteriosclerosis in heart transplantation. However, reliable pieces of evidence are still limited concerning the source of the regenerated endothelium within the transplanted heart.METHODS: We analyzed single-cell RNA sequencing data and constructed an inducible lineage tracing mouse, combined heart transplantation with bone marrow transplantation and a parabiosis model, cellular components, and endothelial cell populations in cardiac graft lesions.RESULTS: Our single-cell RNA sequencing analysis of a transplanted heart allowed for the establishment of an endothelial cell atlas with a heterogeneous population, including arterial, venous, capillary, and lymphatic endothelial cells. Along with genetic cell lineage tracing, we demonstrated that the donor cells were mostly replaced by recipient cells in the cardiac allograft, up to 83.29% 2 weeks after transplantation. Furthermore, recipient nonbone marrow CD34+ endothelial progenitors contributed significantly to extracellular matrix organization and immune regulation, with higher apoptotic ability in the transplanted hearts. Mechanistically, peripheral blood-derived human endothelial progenitor cells differentiate into endocardial cells via Vascular endothelial growth factor receptor-mediated pathways. Host circulating CD34+ endothelial progenitors could repair the damaged donor endothelium presumably through CCL3-CCR5 chemotaxis. Partial depletion of host CD34+ cells resulted in delayed endothelial regeneration.CONCLUSIONS: We created an annotated fate map of endothelial cells in cardiac allografts, indicating how recipient CD34+ cells could replace the donor endothelium via chemokine CCL3-CCR5 inter-actions. The mechanisms we discovered could have a potential therapeutic effect on the long-term outcomes of heart transplantation.J Heart Lung Transplant 2023;42:1651-1665 (c) 2023 International Society for Heart and Lung Transplantation. All rights reserved.

Introduction Diabetes mellitus (DM) is a major public health problem worldwide. Cancer is the second most common cause of death in the United States and the leading cause of death in China. There is compelling evidence that individual risk for type 2 diabetes mellitus (T2DM) is strongly influenced by genetic factors. DM and cancer may interact with one another; some kinds of cancer accompany DM, and DM can also promote cancer.Methods An analysis was conducted of diabetes mellitus-related gene (DM-gene) expression levels in tumor and normal tissues, clinical parameters, tumor stages, mutations, copy number variations (CNVs), immune cell infiltration, survival, gene enrichment, and gene ontology annotations.Results This analysis revealed six genes that appear to play key roles in lung cancer survival: MTMR3 (in lung adenocarcinoma [LUAD]) and COBLL1, PPARG, PPIP5K2, RREB1, and WFS1 (in lung squamous cell carcinoma [LUSC]).Conclusion The results suggested that clinical practitioners and researchers should account for PPARG and RREB1 expression when selecting or testing chemotherapy drugs.

Cardiac fibrosis is a common consequence of various cardiac diseases and is linked to the activation of cardiac fibroblasts (FBs). This study aimed to investigate the effect of tadalafil (Tad) on cardiac fibrosis by regulating FB transformation and its molecular mechanism. Angiotensin II (Ang II) treated FBs were used to construct in vitro cardiac fibrosis models. FBs were divided into three groups: control, Ang II, and Ang II+Tad. Immunofluorescence, real-time quantitative PCR, Western blot, and flow cytometry were utilized to detect the expression of α -SMA, collagen types I and III, fibronectin, Bcl-2, Bax, Bad, TGF- β 1, SMAD3, p-SMAD3, and apoptosis. The expression of α -SMA, collagen types I and III, fibronectin, and Bcl-2 in the Ang II+Tad group was significantly decreased, while the expressions of Bax and Bad were increased compared to the Ang II group. The Ang II+Tad group also had the highest proportion of apoptosis. The TGF- β 1/Smad3 signaling pathway was found to be inhibited in the Ang II+Tad group. Therefore, Tad can inhibit Ang II-induced FB activation through the TGF- β 1/Smad3 pathway and promote apoptosis of activated FBs.

Inhibition of DNA binding proteins 1 and 3 (ID1 and ID3) are important downstream targets of BMP signalling that are necessary for embryonic development. However, their specific roles in regulating the pluripotency of human embryonic stem cells (hESCs) remain unclear. Here, we examined the roles of ID1 and ID3 in primed and naive-like hESCs and showed that ID1 and ID3 knockout lines (IDs KO) exhibited decreased survival in both primed and naive-like state. IDs KO lines in the primed state also tended to undergo pluripotent dissolution and ectodermal differentiation. IDs KO impeded the primed-to-naive transition (PNT) of hESCs, and overexpression of ID1 in primed hESCs promoted PNT. Furthermore, single-cell RNA sequencing demonstrated that ID1 and ID3 regulated the survival and pluripotency of hESCs through the AKT signalling pathway. Finally, we showed that TCF3 mediated transcriptional inhibition of MCL1 promotes AKT phosphorylation, which was confirmed by TCF3 knockdown in KO lines. Our study suggests that IDs/TCF3 acts through AKT signalling to promote survival and maintain pluripotency of both primed and naive-like hESCs.

Background and Objectives:Endothelial progenitor cells (EPCs) and endothelial cells (ECs) have been applied in the clinic to treat pulmonary arterial hypertension (PAH), a disease characterized by disordered pulmonary vasculature. However, the lack of sufficient transplantable cells before the deterioration of disease condition is a current limitation to apply cell therapy in patients. It is necessary to differentiate pluripotent stem cells (PSCs) into EPCs and identify their characteristics.Methods and Results:Comparing previously reported methods of human PSCs-derived ECs, we optimized a highly efficient differentiation protocol to obtain cells that match the phenotype of isolated EPCs from healthy donors. The protocol is compatible with chemically defined medium (CDM), it could produce a large number of clinically applicable cells with low cost. Moreover, we also found PSCs-derived EPCs express CD133, have some characteristics of mesenchymal stem cells and are capable of homing to repair blood vessels in zebrafish xenograft assays. In addition, we further revealed that IPAH PSCs-derived EPCs have higher expression of proliferation-related genes and lower expression of immune-related genes than normal EPCs and PSCs-derived EPCs through microarray analysis.Conclusions:In conclusion, we optimized a highly efficient differentiation protocol to obtain PSCs-derived EPCs with the phenotypic and molecular characteristics of EPCs from healthy donors which distinguished them from EPCs from PAH.

Background: Recent studies have demonstrated the beneficial effects of STS in treating pulmonary hypertension by inhibiting the pulmonary vascular remodeling and suppressing the abnormally elevated proliferation and migration of PASMCs. However, the roles of STS on pulmonary vascular endothelium remain largely known.& nbsp;Methods: In this study, we investigated the effects and mechanisms of STS on pulmonary vascular endothelial dysfunction by using a chronic hypoxia-induced pulmonary hypertension (HPH) rat model, as well as in pri-marily cultured rat PMVECs and human ESC-ECs cell models.& nbsp;Results: Firstly, a 21-day treatment of STS significantly prevents the disease development of HPH by normalizing the right ventricular systolic pressure and right ventricular hypertrophy, improving the cardiac output. Then, STS treatment markedly inhibits the hypoxia-induced medial wall thickening of the distal intrapulmonary arteries. Notably, STS significantly inhibits the hypoxia-induced apoptosis in both the pulmonary endothelium of HPH rats and primarily cultured PMVECs, through the stabilization of BMPR2 protein and protection of the diminished BMP9-BMPR2-Smad1/5/9 signaling pathway. In mechanism, STS treatment retrieves the hypoxic down-regulation of BMPR2 by stabilizing the BMPR2 protein, inhibiting the BMPR2 protein degradation via lysosome system, and promoting the plasma membrane localization of BMPR2, all of which together reinforcing the BMP9-induced signaling transduction in both PMVECs and human ESC-ECs. However, these effects are absent in hESC-ECs expressing heterozygous dysfunctional BMPR2 protein (BMPR2(+/R899X)).& nbsp;Conclusion: STS may exert anti-apoptotic roles, at least partially, via induction of the BMP9-BMPR2-Smad1/5/9 signaling transduction in pulmonary endothelium and PMVECs.

Objectives The aim of this study is to investigate the clinical significance of myeloperoxidase (MPO)–antineutrophil cytoplasmic antibody (ANCA) on eosinophilic granulomatosis with polyangiitis (EGPA) from a longitudinal Chinese cohort. Methods A total of 120 patients with EGPA were consecutively enrolled and followed up. Two patients with PR3 ANCA was excluded and our analysis focused on the 118 patients with EGPA. On the basis of MPO-ANCA status, baseline clinical manifestations, treatment, and outcomes were analyzed. Logistic regression analysis was performed to analyze the independently associated factors for renal involvement. Results ANCA positivity was observed in 24.2% of patients with EGPA. Patients with MPO-ANCA accounted for 20.8%. Patients with positive MPO-ANCA had higher levels of erythrocyte sedimentation rate (ESR), C-reactive protein, Birmingham Vasculitis Activity Score (BVAS), higher ratios of fever, myalgia, renal involvement, and biopsy-proven vasculitis. Heart manifestations and asthma were more common in patients with negative ANCA. Baseline MPO-ANCA titers positively correlated with ESR, eosinophil count, and BVAS and were higher in patients with methylprednisolone pulse. Among patients with renal involvement, patients with positive MPO-ANCA had higher proportions of female, fever, biopsy-proven vasculitis, and faster ESR; patients with negative ANCA developed more skin and cardiac involvement. MPO-ANCA positivity, male, and ear involvement were the independent factors associated with renal involvement. Intravenous cyclophosphamide and immunoglobulins were prescribed more frequently in patients with positive MPO-ANCA. Conclusion In this cohort, patients with positive MPO-ANCA and negative ANCA displayed distinct clinical features, suggesting that MPO-ANCA might be a valuable biomarker for EGPA stratification. Baseline MPO-ANCA level correlated positively with disease activity of EGPA. MPO-ANCA was a significant independent factor associated with renal involvement.