Identification of Potential lncRNA-miRNA-mRNA Regulatory Network Contributing to Arrhythmogenic Right Ventricular Cardiomyopathy.

Arrhythmogenic right ventricular cardiomyopathy (ARVC) can lead to sudden cardiac death and life-threatening heart failure. Due to its high fatality rate and limited therapies, the pathogenesis and diagnosis biomarker of ARVC needs to be explored urgently. This study aimed to explore the lncRNA-miRNA-mRNA competitive endogenous RNA (ceRNA) network in ARVC. The mRNA and lncRNA expression datasets obtained from the Gene Expression Omnibus (GEO) database were used to analyze differentially expressed mRNA (DEM) and lncRNA (DElnc) between ARVC and non-failing controls. Differentially expressed miRNAs (DEmiRs) were obtained from the previous profiling work. Using starBase to predict targets of DEmiRs and intersecting with DEM and DElnc, a ceRNA network of lncRNA-miRNA-mRNA was constructed. The DEM and DElnc were validated by real-time quantitative PCR in human heart tissue. Protein-protein interaction network and weighted gene co-expression network analyses were used to identify hub genes. A logistic regression model for ARVC diagnostic prediction was established with the hub genes and their ceRNA pairs in the network. A total of 448 DEMs (282 upregulated and 166 downregulated) were identified, mainly enriched in extracellular matrix and fibrosis-related GO terms and KEGG pathways, such as extracellular matrix organization and collagen fibril organization. Four mRNAs and two lncRNAs, including COL1A1, COL5A1, FBN1, BGN, XIST, and LINC00173 identified through the ceRNA network, were validated by real-time quantitative PCR in human heart tissue and used to construct a logistic regression model. Good ARVC diagnostic prediction performance for the model was shown in both the training set and the validation set. The potential lncRNA-miRNA-mRNA regulatory network and logistic regression model established in our study may provide promising diagnostic methods for ARVC.

Circulating trimethylamine N-oxide is correlated with high coronary artery atherosclerotic burden in individuals with newly diagnosed coronary heart disease.

BACKGROUND: Trimethylamine N-oxide (TMAO) is a metabolite derived from the gut microbiota and has been reported to be correlated with cardiovascular diseases. Although TMAO is associated with the severity of coronary artery disease in subjects with coronary heart disease (CHD) history. However, the correlation between TMAO and the atherosclerotic burden in newly diagnosed cases of CHD is unknown. METHODS: In this hospital-based study, we enrolled 429 individuals newly diagnosed with CHD undergoing coronary angiography. Plasma TMAO was assessed before coronary angiography. SYNTAX score was computed during coronary angiography to estimate the coronary artery atherosclerotic burden. Both linear and logistic regression analyses were conducted to explore the correlation between plasma TMAO levels and SYNTAX score in newly diagnosed CHD population. RESULTS: The TMAO in patients with SYNTAX ≥ 33 and subjects with SYNTAX < 23 were 6.10 (interquartile range [IQR]: 3.53 to 9.15) µmol/L and 4.90 [IQR: 3.25 to 7.68] µmol/L, respectively. Linear regression adjusting for traditional risk factors showed TMAO level was positively correlated with SYNTAX score (ß = 0.179; p = 0.006) in CHD population. When TMAO was added to models with traditional risk factors, the predictive value improved significantly, with the receiver operating characteristic curve (AUC) increased from 0.7312 to 0.7502 (p = 0.003). Stratified analysis showed that the correlations did not hold true for subjects who were non-smoker or with histories of diabetes. None of the stratifying factors significantly altered the correlation (all p for interaction < 0.05). CONCLUSIONS: We found a positive linear correlation between plasma TMAO and SYNTAX score among newly diagnosed CHD individuals in Chinese population.

In Situ Polymerization Derived from PAN-Based Porous Membrane Realizing Double-Stabilized Interface and High Ionic Conductivity for Lithium-Metal Batteries.

Polymer polyacrylonitrile (PAN), with exceptional mechanical strength and ionic conductivity, is considered a potential electrolyte. However, the huge interfacial impedance of PAN-derived C≡N polar nitrile groups and Li anode limited its application. In this study, a double-stabilized interface was integrated by in situ polymerization of DOL between electrodes and a three-dimensional (3D) porous PAN polymer matrix containing SN plasticizer and LLZTO ceramic fillers to optimize the challenge of interfacial instability. The fabricated PDOL-PAN(SN/LLZTO)-PDOL composite solid electrolyte (CSE) exhibited the maximum ionic conductivities of 1.9 × 10-3 S cm-1 at room temperature and 2.5 × 10-3 S cm-1 at 60 °C, an electrochemical stability window (ESW) of 4.9 V, and a high Li+ transference number (tLi+) of 0.65. In addition, the side reactions of the PAN/Li metal were effectively prevented by inserting PDOL between the 3D porous membrane and Li electrode. Benefiting from the superior interface compatibility and ion conductivity, the Li symmetric battery showed more than 2000 h of cyclability. The solid Li/LiFePO4 full battery delivered excellent cycling performance, showing an original specific capacity of 136.2 mAh g-1 with a capacity retention of 90.1% after 350 cycles at 1C and 60 °C. Furthermore, the cycling of solid-state Li/NCM622 batteries also proved their application potential. This work presents an effective approach to solving interface problems of the PAN electrolyte for solid lithium-metal batteries (LMBs).

PM2.5 associates with blood pressure: a Mendelian randomization analysis.

The relationship between fine particulate matter (PM2.5) and blood pressure (BP) is a controversial issue. We conducted a two-sample Mendelian randomization (MR) analysis and identified 58 genome-wide significant single-nucleotide polymorphisms associated with PM2.5 as instrument variables. Inverse-variance weighted (IVW) was used as the primary analysis approach. MR-Egger, weighted median, simple model, and weighted model methods were selected for quality control. We found a significant negative causal association of higher genetically predicted PM2.5 levels with lower systolic BP (SBP), while no causal relationship was identified between PM2.5 and diastolic BP (DBP). For each 1 standard deviation increase in genetically predicted PM2.5 levels, the beta value (95% CI) of SBP was -0.14 (-0.25, -0.03) for IVW (p=0.02), and -0.13 (-0.22, -0.04) for weighted median (p=0.005). Increased PM2.5 concentrations can lead to decreased SBP levels. Our findings provided novel insights into the controversial topic on the causal relationship between PM2.5 and BP.

PBX/Knotted 1 homeobox-2 (PKNOX2) is a novel regulator of myocardial fibrosis.

Much effort has been made to uncover the cellular heterogeneities of human hearts by single-nucleus RNA sequencing. However, the cardiac transcriptional regulation networks have not been systematically described because of the limitations in detecting transcription factors. In this study, we optimized a pipeline for isolating nuclei and conducting single-nucleus RNA sequencing targeted to detect a higher number of cell signal genes and an optimal number of transcription factors. With this unbiased protocol, we characterized the cellular composition of healthy human hearts and investigated the transcriptional regulation networks involved in determining the cellular identities and functions of the main cardiac cell subtypes. Particularly in fibroblasts, a novel regulator, PKNOX2, was identified as being associated with physiological fibroblast activation in healthy hearts. To validate the roles of these transcription factors in maintaining homeostasis, we used single-nucleus RNA-sequencing analysis of transplanted failing hearts focusing on fibroblast remodelling. The trajectory analysis suggested that PKNOX2 was abnormally decreased from fibroblast activation to pathological myofibroblast formation. Both gain- and loss-of-function in vitro experiments demonstrated the inhibitory role of PKNOX2 in pathological fibrosis remodelling. Moreover, fibroblast-specific overexpression and knockout of PKNOX2 in a heart failure mouse model induced by transverse aortic constriction surgery significantly improved and aggravated myocardial fibrosis, respectively. In summary, this study established a high-quality pipeline for single-nucleus RNA-sequencing analysis of heart muscle. With this optimized protocol, we described the transcriptional regulation networks of the main cardiac cell subtypes and identified PKNOX2 as a novel regulator in suppressing fibrosis and a potential therapeutic target for future translational studies.

Heterogeneous nucleation induced A. pernyi/B. mori silk fibroin coatings on AZ31 biometals with enhanced corrosion resistance, adhesion and biocompatibility.

Silk fibroin coatings on biomedical magnesium alloys have garnered significant attention due to their enhanced corrosion resistance and biocompatibility. However, the utilization of wild A. pernyi silk fibroin, known for its RGD sequence that facilitates tissue regeneration, presents a challenge for corrosion-resistant coatings on magnesium alloys due to its weak adhesion and high dissolution rate. In this study, we employed hexafluoroisopropanol as a solvent to blend A. pernyi silk fibroin with B. mori silk fibroin. The resulting blended fibroin coating at a 3:7 mass ratio exhibited a heterogeneous nucleation effect, enhancing ß-sheet content (32.3 %) and crystallinity (28.6 %). This improved ß-sheet promoted the "labyrinth effect" with an Icorr of 2.15 × 10-6 A cm-2, resulting in significantly improved corrosion resistance, which is two orders of magnitude lower than that of pure magnesium alloy. Meanwhile, the increased content of exposed serine in zigzag ß-sheet contributes to a higher adhesion strength. Cell cytotoxicity evaluation confirmed the enhanced cell adhesion and bioactivity. This work provides a facile approach for wild A. pernyi silk fibroin coatings on magnesium alloys with enhanced corrosion resistance, adhesion and biocompatibility.

Versican Promotes Cardiomyocyte Proliferation and Cardiac Repair.

BACKGROUND: The adult mammalian heart is incapable of regeneration, whereas a transient regenerative capacity is maintained in the neonatal heart, primarily through the proliferation of preexisting cardiomyocytes. Neonatal heart regeneration after myocardial injury is accompanied by an expansion of cardiac fibroblasts and compositional changes in the extracellular matrix. Whether and how these changes influence cardiomyocyte proliferation and heart regeneration remains to be investigated. METHODS: We used apical resection and myocardial infarction surgical models in neonatal and adult mice to investigate extracellular matrix components involved in heart regeneration after injury. Single-cell RNA sequencing and liquid chromatography-mass spectrometry analyses were used for versican identification. Cardiac fibroblast-specific Vcan deletion was achieved using the mouse strains Col1a2-2A-CreER and Vcanfl/fl. Molecular signaling pathways related to the effects of versican were assessed through Western blot, immunostaining, and quantitative reverse transcription polymerase chain reaction. Cardiac fibrosis and heart function were evaluated by Masson trichrome staining and echocardiography, respectively. RESULTS: Versican, a cardiac fibroblast-derived extracellular matrix component, was upregulated after neonatal myocardial injury and promoted cardiomyocyte proliferation. Conditional knockout of Vcan in cardiac fibroblasts decreased cardiomyocyte proliferation and impaired neonatal heart regeneration. In adult mice, intramyocardial injection of versican after myocardial infarction enhanced cardiomyocyte proliferation, reduced fibrosis, and improved cardiac function. Furthermore, versican augmented the proliferation of human induced pluripotent stem cell-derived cardiomyocytes. Mechanistically, versican activated integrin ß1 and downstream signaling molecules, including ERK1/2 and Akt, thereby promoting cardiomyocyte proliferation and cardiac repair. CONCLUSIONS: Our study identifies versican as a cardiac fibroblast-derived pro-proliferative proteoglycan and clarifies the role of versican in promoting adult cardiac repair. These findings highlight its potential as a therapeutic factor for ischemic heart diseases.

Relationship between daytime napping and cardiovascular disease: A two-sample mendelian randomization study.

OBJECTIVE: Daytime napping has been reported to have a potential association with an increased risk of cardiovascular diseases (CVDs) in several cohort studies, but the causal effects are unclear. In this study, we aimed to investigate the relationship between daytime napping and CVDs, as well as to validate causality in this relationship by Mendelian randomization (MR). METHODS: A two-sample MR method was used to evaluate the causal effect of daytime napping on CVDs. The exposure of daytime napping was extracted from publicly available genome-wide association studies (GWASs) in the UK Biobank, and the outcomes of 14 CVDs were obtained from the FinnGen consortium. A total of 49 single-nucleotide polymorphisms (SNPs) were used as the instrumental variables. The effect estimates were calculated by using the inverse-variance weighted method. RESULTS: The MR analyses showed that genetically predicted daytime napping was associated with an increased risk of five CVDs, including heart failure (odds ratio (OR): 1.71, 95% CI: 1.19-2.44, p = 0.003), hypertension (OR: 1.51, 95% CI: 1.05-2.16, p = 0.026), atrial fibrillation (OR: 1.71, 95% CI: 1.02-2.88, p = 0.042), cardiac arrythmias (OR: 1.47, 95% CI: 1.47, 95% CI: 1.01-2.13, p = 0.042) and coronary atherosclerosis (OR: 1.77, 95% CI: 1.17-2.68, p = 0.006). No significant influence was observed for other CVDs. CONCLUSION: This two-sample MR analysis suggested that daytime napping was causally associated with an increased risk of heart failure, hypertension, atrial fibrillation, cardiac arrythmias and coronary atherosclerosis.

In Situ Gelation of a 1,3-Dioxolane Dual-Permeable Porous Tandem Framework with Excellent Interfacial Stability to Power Long-Cycling Solid-State Lithium Metal Batteries.

Ceramic Li1.3Al0.3Ti1.7(PO4)3 (LATP) with high ionic conductivity and stability in ambient atmosphere is considered to be potent as a solid-state electrolyte of solid-state lithium metal batteries (SSLMBs), but its huge interfacial impedance with electrodes and the unwanted Ti4+-mediated reduction reaction caused by the lithium (Li) metal anode greatly limit its application in LMBs. Herein, a composite polymer electrolyte (CPET) was integrated by in situ gelation of dual-permeable 1, 3-dioxolane (DOL) in the tandem framework composed of the commercial cellulose membrane TF4030 and a porous three-dimensional (3D) skeleton-structured LATP. The in situ gelled DOL anchored in the tandem framework ensured nice interfacial contact between the as-prepared CPET and electrodes. The introduction of the porous 3D LATP endowed CPET the increased lithium-ion migration number (tLi+) of 0.70, a wide electrochemical stability window (ESW) of 4.86 V, and a high ionic conductivity of 1.16 × 10-4 S cm-1 at room temperature (RT). Meanwhile, the side reaction of the LATP/Li metal was adequately restrained by inserting TF4030 between the porous LATP and Li anode. Profiting from the superb interfacial stability and the enhanced ionic transport capacity of CPET, Li/Li batteries based on the optimal CPET (CPET2) cycled over 2000 h at 20∼30 °C smoothly. Moreover, solid-state LiFePO4 (LFP)/Li with CPET2 exhibited excellent electrochemical performance with a capacity retention ratio of 72.2% after 400 cycles at 0.5C. This work offers an integrated strategy to guide the fabrication of a highly conductive solid electrolyte and a stable interface design for high-performance SSLMBs.

Rapid warning of emerging contaminants in reuse water using biocathode sensors.

The proliferation of emerging contaminants (ECs) in the environment poses a major threat to the safety of reuse water. However, many ECs exist for which no corresponding control standards have been established. Here, we used polarity reversal to construct a biocathode sensor capable of early warning of ECs biotoxicity in aerobic reuse water with low organic concentrations. The baseline current and sensitivity of the biosensor in response to formaldehyde were enhanced by 25% and 23% using microbial fuel cell effluent as the inoculum. The microbial community explained that the inoculum primarily influenced the performance of the biosensor by modulating species abundance, function and interactions. More importantly, the successfully commissioned biocathode sensor demonstrated rapid warning capability (Response time less than 1.3 h) for ECs such as fluoride, disinfection by-products and antibiotics in an actual landscape reuse system. Further, the sensor could quantify the concentration of a single known contaminant. Our study demonstrated a method for rapid early warning of ECs in an oxygen-rich, low-organics environment, promoting innovative development of monitoring technologies for water ecology and environmental safety.

Full Life-Cycle Lead Management and Recycling Transparent Conductors for Low-Cost Perovskite Solar Cell.

Realizing a full life-cycle management for toxic lead (Pb) and reducing material/manufacture cost are the key steps in determining the commercialization process of perovskite photovoltaics. In this work, we develop full lifecycle material management for a carbon-based perovskite solar cell (C-PSC) to immobilize and recover Pb against environmental pollution, followed by refabrication of C-PSC based on recovered materials and recycled transparent conductors from obsolete devices. Pb immobilization is first achieved by a strong coordination interaction between undercoordinated Pb ions from perovskite and a C═O bond from green pseudohalide ions (pseudo-X), and the resulting C-PSC with the structure of ITO/SnO2/pseudo-X-perovskite/carbon yields an efficiency of 16.63%. Pb from an end-of-life C-PSC is then recovered by dissolving the obsolete perovskite layer into DMF/DMSO precursor solvent, followed by replenishing a certain amount of MAI to guarantee new perovskite layer formation. The refabricated C-PSC based on recovered perovskite and a recycled transparent conductor displays comparable efficiency (15.30%) to that of C-PSC with commercial raw materials, also exceeding the previous efficiency record for C-PSCs based on recycled materials. Such refabricated C-PSC is relatively low-cost.

Identification of potential lncRNA-miRNA-mRNA regulatory network contributing to aldosterone-producing adenoma.

Aldosterone-producing adenoma (APA) is a common cause of secondary hypertension. This study aimed to explore the lncRNA-miRNA-mRNA competitive endogenous RNA (ceRNA) network to uncover molecular mechanism underlying APA. The mRNA and lncRNA expression data of APA and adjacent adrenal gland (AAG) from GSE60044, GSE64957 and GSE101894 were obtained from the Gene Expression Omnibus (GEO) database to analyse differentially expressed genes (DEGs) and lncRNAs (DElncs). Hub genes were identified by robust rank aggregation (RRA) and protein-protein interaction (PPI) network analysis. The miRcode and miRWalk network tools were used to construct the ceRNA network. 1526 upregulated and 1512 downregulated DEGs were identified, which are mainly enriched in extracellular matrix and Ca2 + -related GO terms. In the KEGG pathway analysis, Ca2+ signalling and the aldosterone synthesis and secretion pathways were enriched. ceRNA network included 2 lncRNAs, 9 miRNAs, and 13 mRNAs. The lncRNAs are MEG3 and LINC00115. The mRNAs included CCND1, TP53, GPRC5B, BMI1, COMMD3-BMI1, ADAMTS15, STAT3, MMP2, SCN2B, CXCL12, HGF, FOS, and THBS1. Overall, this study conducted a ceRNA regulatory network analysis and identified that 2 lncRNAs and 13 mRNAs may contribute to the development of APA. These findings may provide novel diagnostic and intervention targets for APA.

The Study of Mechanical Behaviors of Caprinae Horn Sheath under Pendulum Impact.

As a light-weight natural keratin biocomposite, Bovidae horn exhibits high mechanical properties and energy absorption. Different to the widely studied horn from subfamily Bovinae and Antilocapridae, few studies have focused on the horn sheath of subfamily Caprinae. In this work, three Caprinae horn sheathes from Cashmere goat, White goat and Black sheep were selected. Charpy pendulum impact tests were performed, and the fracture characteristics were evaluated. It was demonstrated that water plays an important role in acquiring balanced dynamic mechanical properties in all Caprinae horn sheaths. The hydrated keratin provides large plastic deformation capacity and further gives rise to a gradual generation of micro-cracks. Multi-scale structure including wavy-shaped interface, scattered voids and hierarchical micro-fibre were observed. Such a structure induced complex fracture mechanisms, such as delamination, 90° crack deflection and fibre pull-out, which were probably influenced by interfacial strength. The results are expected to endow the research and thinking of Bovidae horn.

Extracellular matrix-based biomaterials for cardiac regeneration and repair.

The spectrum of ischemic heart diseases, encompassing acute myocardial infarction to heart failure, represents the leading cause of death worldwide. Although extensive progress in cardiovascular diagnoses and therapy has been made, the prevalence of the disease continues to increase. Cardiac regeneration has a promising perspective for the therapy of heart failure. Recently, extracellular matrix (ECM) has been shown to play an important role in cardiac regeneration and repair after cardiac injury. There is also evidence that the ECM could be directly used as a drug to promote cardiomyocyte proliferation and cardiac regeneration. Increasing evidence supports that applying ECM biomaterials to maintain heart function recovery is an important approach to apply the concept of cardiac regenerative medicine to clinical practice in the future. Here, we will introduce the essential role of cardiac ECM in cardiac regeneration and summarize the approaches of delivering ECM biomaterials to promote cardiac repair in this review.

Optimized Langendorff perfusion system for cardiomyocyte isolation in adult mouse heart.

With the rapid development of single-cell sequencing technology, the Langendorff perfusion system has emerged as a common approach to decompose cardiac tissue and obtain living cardiomyocytes to study cardiovascular disease with the mechanism of cardiomyocyte biology. However, the traditional Langendorff perfusion system is difficult to master, and further, the viability and purity of cardiomyocytes are frequently unable to meet sequencing requirements due to complicated devices and manipulate processes. Here, we provide an optimized Langendorff perfusion system with a simplified and standardized operating protocol which utilizes gravity as the perfusion pressure, includes a novel method for bubbles removing and standardizes the criteria for termination of digestion. We obtained stable cardiomyocyte with high viability and purity after multiple natural gravity sedimentation. The combination of the optimized Langendorff perfusion system and the multiple natural gravity sedimentation provides a stable system for isolating adult mouse heart, which will provide higher-quality cardiomyocytes for further experiments.

gp130 Controls Cardiomyocyte Proliferation and Heart Regeneration.

BACKGROUND: A key cause of the high mortality of cardiovascular diseases is the cardiomyocyte inability to renew after cardiac injury. As a promising strategy to supplement functional myocytes for cardiac repair, there is a pressing need to understand the cellular and molecular mechanisms of heart regeneration. METHODS: Seven genetic mouse lines were used: global OSM (oncostatin M) knockout, monocyte-/macrophage-specific OSM deletion, cardiomyocyte-specific lines, including OSM receptor deletion, gp130 (glycoprotein 130) deletion, gp130 activation, and Yap (yes-associated protein) ablation with gp130 activation mice. A series of molecular signaling experiments, including RNA sequencing, immunostaining, coimmunoprecipitation, and imaging flow cytometry, were conducted. Two models of cardiac injury, apical resection and myocardial infarction operation, were performed in neonatal, juvenile, and adult mice. Heart regeneration and cardiac function were evaluated by Masson staining and echocardiography, respectively. Gene recombinant adenovirus-associated virus was constructed and infected myocardial-infarcted mice as a gene therapy. RESULTS: OSM was identified by RNA sequencing as a key upstream regulator of cardiomyocyte proliferation during neonatal heart regeneration in mice. Cardiomyocyte proliferation and heart regeneration were suspended in neonatal mice after cardiac injury when OSM was conditionally knockout in macrophages. The cardiomyocyte-specific deficiency of the OSM receptor heterodimers, OSM receptor and gp130, individually in cardiomyocytes reduced myocyte proliferation and neonatal heart regeneration. Conditional activation of gp130 in cardiomyocytes promoted cardiomyocyte proliferation and heart regeneration in juvenile and adult mice. Using RNA sequencing and functional screening, we found that Src mediated gp130-triggered cardiomyocyte proliferation by activating Yap (yes-associated protein) with Y357 phosphorylation independently of the Hippo pathway. Cardiomyocyte-specific deletion of Yap in Myh6-gp130ACT mice blocked the effect of gp130 activation-induced heart regeneration in juvenile mice. Gene therapy with adenovirus-associated virus encoding constitutively activated gp130 promoted cardiomyocyte proliferation and heart regeneration in adult mice after myocardial infarction. CONCLUSIONS: Macrophage recruitment is essential for heart regeneration through the secretion of OSM, which promotes cardiomyocyte proliferation. As the coreceptor of OSM, gp130 activation is sufficient to promote cardiomyocyte proliferation by activating Yap through Src during heart regeneration. gp130 is a potential therapeutic target to improve heart regeneration after cardiac injury.

The microfabrication of mold for polymer microfluidic devices with Zr-based metallic glass.

Polymer microfluidic devices are used for many purposes such as microarrays and biochips. The key tool for manufacturing these chips in bulk is an appropriate mold. However, the popular material for making molds is nickel or nickel alloys, which have low stiffness and wear out easily. Zr-based metallic glass is a promising material for micro- or nanomolds because it has good mechanical properties and can be easily formed with high precision. In this paper, Zr-based metallic glass is proposed for use as micromold insert to make poly-(methyl methacrylate) (PMMA) microfluidic devices. Our experiments show that they have good feature integrity and replication quality. Microchannels we fabricated using these replicas did not leak and had good flow performance. Zr-based metallic glass can greatly ease the manufacture of plastic microfluidic devices for research and commercial applications.