Congenital diaphragmatic hernia (CDH) is a severe developmental anomaly with high etiological heterogeneity. Although numerous susceptibility genes have been identified, genetic factors alone fail to explain the full disease risk. Epidemiological data highlight the contribution of environmental exposures. Epigenetic regulation provides a crucial link bridging genetic predisposition and environmental influence. This review aims to summarize recent multiomics evidence elucidating how epigenetic mechanisms mediate gene-environment interactions in CDH. This narrative review summarizes studies published over the past decade on epigenetic mechanisms in congenital diaphragmatic hernia (CDH), including DNA methylation, histone modifications, noncoding RNAs, and integrative genomic and transcriptomic analyses in human samples, animal models, and iPSC-derived organoids. Relevant literature was identified through targeted searches of the biomedical literature and screening of reference lists from key articles. Relevant studies were considered according to their relevance to epigenetic regulation, gene-environment interactions, and translational implications in CDH. Aberrant DNA methylation, histone acetylation imbalance, and dysregulated miRNAs converge on key developmental pathways, including retinoic acid, TGF-β, and NF-κB signaling. Experimental evidence shows that miR-200b supplementation or pharmacological restoration of histone acetylation can partially rescue pulmonary hypoplasia in nitrofen-induced CDH models. iPSC and organoid systems further demonstrate synergistic effects between genetic susceptibility and mechanical stress, supporting epigenetic regulation as a mechanistic bridge. Epigenetic mechanisms serve as core mediators linking genetic and environmental factors in CDH pathogenesis. Future directions include large-scale EWAS/WGBS and multiomics integration, establishment of standardized epigenetic databases, and ethically guided development of prenatal epigenetic diagnostics and targeted interventions.
Methicillin-resistant Staphylococcus aureus (MRSA) is a major antimicrobial-resistant pathogen affecting both human and animal health. Although historically associated with healthcare settings, MRSA is now established in livestock production and throughout the production chain. Its detection in animals, food products, and processing environments reflects the complex ecology of antimicrobial resistance (AMR) in modern food systems. This narrative review synthesizes current evidence on the molecular basis of methicillin resistance and multidrug resistance determinants, as well as the epidemiology of MRSA in food-associated settings. Particular emphasis is placed on its occurrence in animal-derived foods and key reservoirs within farms, slaughterhouses, and processing environments. Livestock-associated populations are dominated by clonal complex CC398. In contrast, CC9 is prevalent in pig production systems in Asia, while CC5-related lineages occur at the human and animal interface. MRSA has been detected in retail meat and animal-derived foods at low but measurable prevalence, indicating contamination during slaughter and processing. Virulence determinants include staphylococcal enterotoxins linked to food poisoning and Panton-Valentine leukocidin associated with severe infections. Biofilm formation and adhesins further support persistence and colonization. Epidemiological and molecular evidence indicates that livestock, processing environments, and food-contact surfaces act as interconnected reservoirs sustaining MRSA circulation. Human exposure occurs primarily through occupational contact and environmental pathways, whereas foodborne transmission appears less common. Effective control requires integrated surveillance, responsible antimicrobial use in livestock production, and strict hygiene practices throughout the production chain within a One Health framework.
This study seeks to investigate the underlying mechanism of glycolytic key gene bisphosphoglycerate mutase (BPGM) in nonalcoholic fatty liver disease (NAFLD). qRT-PCR and immunohistochemistry were utilized to detect BPGM levels in clinical NAFLD samples. HepG2 cells and liver organoids were treated with free fatty acid. (FFA). The role of BPGM in NAFLD was explored at cellular, organoid, and animal levels. Metabolomics was performed to analyze differential metabolites and metabolic pathways. Furthermore, we examined the regulatory mechanisms of BPGM by HIF-1α in NAFLD. Results indicated that high expression of BPGM in NAFLD samples was correlated with NAFLD progression. Moreover, Severe group had higher BPGM expression than Mild group. FFA treatment induced time-dependent steatosis and BPGM upregulation in HepG2 cells and liver organoids, whereas BPGM knockdown attenuated lipid accumulation, cellular injury, and oxidative stress. At the animal level, knockdown of BPGM reversed high-fat diet (HFD) induced lipid accumulation and liver tissue injury. Metabolomics studies showed significant changes of metabolic pathways including glycolysis/gluconeogenesis and pyruvate metabolism. Verification experiment showed FFA increased pyruvic acid levels, and knockdown of BPGM decreased pyruvic acid levels. Pyruvic acid further reversed the changes in NAFLD progression caused by BPGM knockdown at the cellular and organoid levels. Finally, HIF-1α regulated the expression of BPGM in NAFLD. Together, our findings suggest that BPGM contributes to abnormal glucose metabolism and promotes hepatic steatosis, thereby driving NAFLD progression.
Modified citrus pectin (MCP), a polysaccharide from citrus fruits, modulates galectin-3 (Gal-3) and immune responses. Although MCP exhibits notable immunomodulatory effects, its role in drug-induced splenic and systemic toxicity remains unexplored. This study investigated the effects of MCP modulation on splenic immune remodeling and galectin expression in a rat model of cisplatin-induced toxicity. Wistar rats were divided into four groups (n = 5 animals/group): control (SHAM), MCP (100 mg/kg/day for 7 days), cisplatin (CIS, 10 mg/kg/day for 3 days), and MCP + CIS. Spleens were collected 6 h after the final cisplatin dose. Cisplatin induced splenic structural disorganization and selectively reduced nitric oxide levels without broadly affecting antioxidant enzymes. Cisplatin treatment was associated with increased CD3⁺ T cell labeling and enhanced tissue expression of Gal-1, -3, and - 9. MCP administration did not restore splenic architecture but promoted increased hemosiderin deposition in the red pulp, suggestive of enhanced erythrophagocytosis and altered iron handling, and markedly reduced CD3⁺ T cell immunoreactivity. MCP associated with cisplatin also reduced Gal-1 and Gal-3 levels and altered the relationship between galectins and splenic immune cell populations. Correlation analyses revealed positive associations between Gal-1, -3, and - 9 and CD68⁺ macrophages, as well as a selective association between Gal-3 and CD3⁺ T cells, exclusively in MCP + CIS animals. MCP does not mitigate cisplatin-induced splenic damage but alters immune cell distribution and galectin-related responses during cisplatin exposure.
Fibromyalgia, a syndrome characterized by hyperalgesia and 'negative emotionality', and major depressive disorder (MDD) demonstrate substantial overlaps in clinical, neurobiological, and therapeutic domains. Currently, treatment options for fibromyalgia remain limited; however, the epidemiology of this syndrome continues to grow worldwide. The use of animal models is indispensable for developing new treatment strategies for fibromyalgia. Meanwhile, the choice of animal paradigms is limited. Here, we used the ultrasound exposure of emotional stress on CBA, BALB/c, and C57BL/6 mouse strains to model this condition and to identify new molecular targets of fibromyalgia treatment. We exposed young male mice of three common strains to a three-week ultrasound stress (US) comprising emotionally negative and neutral frequencies of 20-25 kHz and 25-45 kHz, resulting in the development of altered pain sensitivity and signs of 'negative emotionality'. Specifically, mice were studied for timid-like/aggressive behaviors and the tail flick response. Serum levels of corticosterone, cortisol, β-Endorphin, and brain-derived neurotrophic factor (BDNF), as well as brain gene expression of interleukin-33 (Il-33), Bdnf, and its receptor Trkb were investigated. Among the stressed mouse strains, C57BL/6 mice displayed augmented pain sensitivity, allodynia, and suppressed dominant behavior, whereas CBA and BALB/c mice demonstrated opposing changes. Glucocorticoid levels were increased in all stressed groups. Stressed C57BL/6 mice showed downregulated gene and protein expression of functionally inter-related BDNF and IL-33 molecules in the hippocampus, amygdala, and striatum, significantly correlating with behavioral outcomes, as well as lowered blood levels of β-Endorphin and elevated cortisol concentrations. Altogether, our study identified the BDNF/IL-33 regulatory pathway as a molecular correlate of fibromyalgia, and the use of US-exposed young C57BL/6 mice as a potential model that recapitulates this syndrome.
Osteoporosis is a metabolic bone disease characterized by reduced bone mass and deterioration of bone microarchitecture, in which impaired osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) represents a central pathological mechanism. In recent years, ferroptosis, a newly recognized form of regulated cell death, has been demonstrated to play an important role in the initiation and progression of osteoporosis. Mitochondrial dysfunction can exacerbate oxidative stress and disrupt iron metabolism, thereby triggering ferroptosis in BMSCs and ultimately inhibiting osteogenic differentiation. The present study aimed to identify and validate key genes associated with mitochondrial homeostasis and osteoporosis, with a particular focus on the role of glial cell line-derived neurotrophic factor (GDNF) in regulating mitochondrial function, suppressing ferroptosis, and promoting osteogenic differentiation of BMSCs. Gene expression data from normal human bone tissues and osteoporotic bone tissues were obtained from public transcriptomic datasets in the Gene Expression Omnibus (GEO) database. Through differential expression analysis, GDNF was identified as a candidate gene. In vitro experiments demonstrated that GDNF markedly improved mitochondrial membrane potential, reduced intracellular reactive oxygen species (ROS) levels, and restored GPX4 expression, thereby promoting osteogenic differentiation of BMSCs. Furthermore, animal experiments confirmed that GDNF intervention effectively increased bone mineral density and improved trabecular microarchitecture in ovariectomized (OVX) mice. In conclusion, this study identified GDNF may suppress ferroptosis by maintaining mitochondrial homeostasis in BMSCs, thereby enhancing osteogenic differentiation and alleviating osteoporosis, providing a potential theoretical basis for molecular targeted therapy in osteoporosis.
Hypertension is the leading global risk factor for cardiovascular diseases, and its pathogenesis is closely linked to excessive sympathetic activation, which markedly elevates the risk of stroke, heart failure and other adverse cardiovascular events. Traditional therapies mainly target peripheral mechanisms, whereas the clinical efficacy of renal denervation highlights the critical role of central regulation in sympathetic hyperactivity. This review focuses on the core sympathetic nuclei including the rostral ventrolateral medulla (RVLM) and paraventricular nucleus (PVN), with epigenetic regulation as a key innovative perspective. We systematically summarize the upstream driving effects of reactive oxygen species (ROS) and neuroinflammation, and emphasize lncRNA/miRNA-mediated post-transcriptional regulation and the modulatory actions of gasotransmitters. Under stress conditions, aberrant activation of ROS and neuroimmune pathways, epigenetic reprogramming, and hyperexcitability of central sympathetic neurons act as key events in sympathetic overactivation, which interact synergistically to promote hypertension. Integrating evidence from multiple hypertensive animal models and clinical studies, we discuss multimodal interventions including pharmacotherapy, nanozyme biotechnology and neuromodulation, analyze current translational challenges, and provide a theoretical framework for developing central-targeted antihypertensive therapies.
The river buffalo is an economically important livestock species supplying milk and meat. However, a multi-tissue transcriptomic atlas for the key dairy river buffalo breeds, Murrah and Nili-Ravi, has not yet been established, and the lack of stable reference genes has hindered in-depth studies of their biological functions and the molecular mechanisms underlying key economic traits such as lactation. We established a multi-tissue gene expression atlas across 20 tissues and identified 717 housekeeping genes (HKGs), and RPL37A and EEF2 were further shown to be stable candidate reference genes under the conditions tested. We found 8368 tissue-specific genes (TSGs), predominantly enriched in the reproductive system. Exploratory analysis of mammary tissue (dry-period vs. public lactating samples, confounded by batch effects) revealed mammary-enriched hub genes including LALBA; these findings are preliminary and require validation. Dynamic analysis across lactation stages (early, peak, mid-, late) identified candidate genes including SEC14L2 and ACSM3. Phenotypic data showed strong negative correlations between milk yield and protein/fat content, and a positive correlation with lactose content. However, causal or regulatory roles were not inferred due to lack of paired individual-level data. Cross-dataset comparisons are descriptive only, and are not key conclusions. In summary, this study lays the foundation for advancing research in lactation trait genetics and functional genomics in river buffalo, with novel reference genes and lactation stage-specific transcriptional dynamics as its main contributions.
Galectins are a functionally diverse family of β-galactosyl-binding lectins that are ubiquitously present in animal species, with key roles in development and immune regulation. Recently, galectins have been found to recognize microbial glycosylated moieties, but the detailed mechanisms of their innate immune functions in mucosal epithelia have remained elusive. The zebrafish (Danio rerio) represents an ideal genetically tractable model to address these questions, as the skin, gills, and gut display mucosal surfaces exposed to the environment. In this study, we investigated the range of endogenous and microbial glycans that are recognized by zebrafish galectin Drgal1 present in epidermal mucus, which would be consistent with defense functions against a bacterial challenge. Results revealed that zebrafish galectin isoform Drgal1-L2 can recognize selected bacterial glycans, as well as zebrafish mucus glycans and cell-surface receptors for bacterial adhesins such as fibronectin (KD = 1.593 × 10-6 M) and CD147 (KD = 1.115 × 10-6 M). Furthermore, preliminary experiments revealed that Drgal1-L2 may hinder bacterial adhesion to epidermal mucus in about 50% at 2.5 μg/mL. Our results suggest that Drgal1-L2 present in epidermal mucus can prevent access of pathogenic bacteria to the epithelial cell surface by alternate or synergic binding to bacterial glycans and to zebrafish mucus components and epithelial receptors for bacterial adhesins. Thus, the present study provides key information for the testing of the abovementioned hypothesis by implementing gene-silencing approaches targeting both zebrafish Drgal1-L2 and its ligands.
Metabolic inflammation, triggered by obesity is a core pathological link in complications such as type 2 diabetes, cardiovascular diseases, and non-alcoholic fatty liver disease. This review explains the central hub role of microRNAs in obesity-related chronic low-grade inflammation, achieved through their precise regulation of macrophage polarisation, inflammatory signalling pathways, and adipokine secretion. Importantly, this relationship is bidirectional-while miRNAs regulate inflammatory cascades, the inflammatory environment itself can alter miRNA expression, forming complex feedback loops. Circulating miRNAs, due to their stability (conferred by encapsulation in exosomes or binding to proteins), tissue specificity, and strong correlation with pathological processes, show great potential as promising non-invasive biomarkers for metabolic inflammation, useful for assessing its activity and complication risk. Furthermore, miRNA-based intervention strategies have demonstrated significant metabolic improvements in animal models. However, their clinical translation still faces challenges such as insufficient delivery targeting, off-target effects, and pharmacokinetic defects. This article aims to summarise the regulatory network of miRNAs in metabolic inflammation and discuss in depth their potential and the bottlenecks to be solved for moving from basic research to clinical application, providing new theoretical basis and strategic directions for the diagnosis and targeted therapy of obesity-related metabolic diseases.
Ulcerative colitis (UC) is a chronic inflammatory bowel disease characterized by dysbiosis of the gut microbiota. Helminth infections are known to modulate host immunity and intestinal microbial composition; however, the therapeutic use of live parasites poses safety challenges. The recombinant Fasciola hepatica fatty acid-binding protein Fh15 is a helminth-derived molecule with anti-inflammatory effects in models of septic shock and dextran sulfate sodium (DSS)-induced colitis. Whether Fh15 also influences gut microbial composition during colitis remains unknown. Male C57BL/6 mice received 4% DSS in drinking water for 7 days to induce colitis and were treated intraperitoneally with Fh15 (2 mg/kg) on days 1, 3, and 5. Fecal samples were collected on days 2, 4, and 7 for 16S rRNA gene sequencing. Standard microbiota pipelines were used to evaluate community diversity. Acute DSS treatment disrupted gut microbial diversity and community structure compared with non-colitic controls. Fh15 treatment partially restored early microbial balance by shifting microbial composition toward that of healthy mice and reducing microbial dispersion, indicating enhanced community stability despite severe dysbiosis. Although alpha diversity did not return to control levels, Fh15 mitigated the expansion of pro-inflammatory genera (Enterococcus and Turicibacter) and preserved beneficial taxa, including Adlercreutzia.
Copy number variations (CNVs) are a major source of structural genomic diversity that influences adaptation, reproduction, and production traits in livestock. The Black Bengal goat, an economically important Indian breed known for its high fecundity, superior skin quality, and resilience to humid tropical climates, was studied to uncover its structural genomic landscape. We performed whole-genome CNV analysis using high-depth (10×) sequencing data from eight individuals. A total of 31,816 copy number variants (CNVs) were identified, predominantly duplications, with an average length of approximately 45 kb. These CNVs were combined into 8910 copy number variation regions (CNVRs) covering approximately 0.15 Gb (about 5.3% of the autosomal genome). CNVR hotspots were mainly located on chromosome 1. Gene annotation showed that regions overlapping with CNVs and CNVRs contained more than 1987 protein-coding genes involved in pathways related to immunity, reproduction, metabolism, and extracellular matrix (ECM) organization. The presence of CNVs involving genes such as GDF9 and BMPR1B on chromosomes 7 & 6, respectively, is important because it indicates that the breed has a high reproductive capacity due to dosage-sensitive duplications. Changes in the extracellular matrix and increased dermal strength have been linked to duplications of genes such as COL6A1, LAMC2, LAMB3, FMN1, and CLDN1. This helps explain the superior hide quality of the breed. This research offers a comprehensive map of CNVs and CNVRs within the genome of the Black Bengal goat. It demonstrates how these duplications lead to structural changes that enhance both reproductive performance and skin resilience. These findings provide a valuable genomic resource for future marker-assisted selection, comparative genomics, and conservation breeding programs aimed at preserving indigenous goat populations.
Alzheimer's disease (AD) research has primarily focused on amyloid beta (Aβ) and tau protein; however, drug development targeting these two proteins has been disappointing. Therefore, there is an urgent need to explore the novel pathogenic mechanisms underlying AD. Recently, we found that expression of the K670N/M671L-mutated amyloid precursor protein (APP) in 293T cells significantly reduced membrane ferroportin (FPN) levels. Furthermore, 2-month-old APP/PS1 mice exhibited a marked decrease in membrane FPN levels, while total FPN expression and Aβ levels remained unchanged. Further studies revealed that features of ferroptosis were present in the brains of 2-month-old APP/PS1 mice, and that treatment with ferroptosis inhibitors or iron chelation significantly alleviated early pathological changes and cognitive impairment in these animals. In addition, supplementation with an APP-FPN binding peptide during the early phase ameliorated AD-related pathologies, including Aβ deposition, neuroinflammation, oxidative stress, and synapse-associated protein deficits, in APP/PS1 mice. Collectively, our findings suggest that APP mutations may contribute to early brain pathological changes and subsequent memory impairment in AD by downregulating membrane trafficking of FPN and inducing ferroptosis, thereby providing new molecular targets for drug development.
Ischemic heart disease is a leading cause of death worldwide. While percutaneous coronary intervention (PCI) restores blood flow in acute coronary syndrome (ACS), reperfusion injury exacerbates myocardial damage, contributing to heart failure (HF). Preemptive administration of a cardioprotective agent could help counter the imminent proinflammatory insult of PCI and reperfusion. Administering BT2, a small-molecule MAPK kinase/extracellular signal-regulated kinase inhibitor, 24 hours before and during ischemia in rats before reperfusion reduced infarct size by ~70% and preserved cardiac function 24 hours and 2 weeks postinjury. BT2 prevented adverse left ventricular remodeling and scarring. Single nucleus RNA sequencing (snRNA-seq) and bulk RNA-seq revealed that BT2 modulated genes associated with inflammation, fibrosis, and matrix production, especially within macrophages and myofibroblasts. BT2 suppressed macrophage and neutrophil infiltration. BT2 reduced the expression of genes in rodent hearts predictive of HF in patients with ACS, including many encoding cytokines, inflammasome components, and damage-associated molecular patterns. BT2 is a small molecule that can prevent myocardial ischemia-reperfusion injury, improve heart function, reduce cardiac fibrosis, and favorably modulate multiple key genes and biological processes in rats prognostic of HF when delivered before reperfusion. This strategy could be evaluated with high-risk unstable angina/non-ST-segment elevation myocardial infarction patients or those having an elective PCI.
Although the anti-inflammatory role of tumor necrosis factor alpha-induced protein 3 (TNFAIP3) in M2 macrophages has been reported, its function in subretinal fibrosis remains unclear. This study aimed to investigate the role of TNFAIP3 in M2 macrophages in subretinal fibrosis and to elucidate its mechanism. A model of subretinal fibrosis of choroidal neovascularization (CNV) lesions was established in C57BL/6 mice using a two-stage laser photocoagulation protocol. TNFAIP3 of M2 macrophages in subretinal fibrosis of CNV lesions was analyzed by single-cell RNA sequencing (scRNA-seq), immunofluorescence, and western blot. In vitro, RAW264.7 cells polarized to M2 phenotype using IL-4 (40 ng/mL) and IL-13 (40 ng/mL) were co-cultured with primary mouse retinal pigment epithelial cells. The effects of Tnfaip3 genetically modulated in M2 macrophages on epithelial-mesenchymal transition and the mechanisms in M2 macrophages were analyzed by immunofluorescence, western blot, and chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR). Peripheral blood monocyte-derived macrophages in mice were depleted through intraperitoneal administration of clodronate liposomes. Subsequently, intravitreal injection of M2 macrophages, M2 macrophages with Tnfaip3-overexpression, or PBS was performed, and the areas of CNV and subretinal fibrosis of CNV lesions were assessed by imaging examinations and immunofluorescence staining. Our findings revealed a transient upregulation followed by subsequent downregulation of TNFAIP3 in M2 macrophages in subretinal fibrosis of CNV lesions and in M2 polarized macrophages. Mechanistically, TNFAIP3 of macrophages inhibits M2 polarization by promoting degradation of nuclear factor kappa B (NF-κB), which reduces Spi1 and subsequently suppresses CCAAT/enhancer-binding protein beta (C/EBPβ). Macrophage depletion led to a significant reduction in both the area of CNV and subretinal fibrosis, with areas measuring (2.23 ± 0.22) × 104 µm2 versus (0.11 ± 0.02) × 104 µm2 for CNV and (6.04 ± 0.64) × 104 µm2 versus (1.17 ± 0.15) × 104 µm2 for fibrosis. Furthermore, intravitreal injection of Tnfaip3-overexpressing M2 macrophages in macrophage-depleted mice attenuated the area of CNV and subretinal fibrosis compared with control M2 macrophages, with areas measuring (0.62 ± 0.08) × 104 µm2 versus (0.11 ± 0.02) × 104 µm2 for CNV and (2.12 ± 0.20) × 104 µm2 versus (1.30 ± 0.17) × 104 µm2 for fibrosis. Our findings demonstrated that TNFAIP3 in macrophages modulates subretinal fibrosis of CNV lesions by regulating M2 polarization through the NF-κB/Spi1/C/EBPβ signaling pathway. These results suggest that targeting TNFAIP3 in M2 macrophages might serve as a promising therapeutic strategy for CNV and subretinal fibrosis.
Myxosporean parasites have been identified in amphibians globally; however, in Japan, no amphibian-infecting myxosporeans have been formally recorded. Thus, in this study, 20 native Japanese frog species were surveyed to clarify the diversity of myxosporeans that infect amphibians. A survey of 102 anurans representing 20 species across six families collected in Japan revealed the presence of a myxosporean belonging to the genus Sphaerospora that parasitizes the kidneys of the Japanese rice frog, Fejervarya kawamurai. The myxosporean is described herein as a new species based on its distinct morphological characteristics and phylogenetic position. Spores are spherical to subspherical with an overall dimension of 11.1 (10.2-12.2) × 11.5 (9.3-14.0) µm. The polar capsules are spherical, with a diameter of 3.3 (2.5-4.3) µm. Typical spores possess a characteristic caudal appendage. This discovery is considered the first record of anuran-parasitic Sphaerospora in East Asia and the fifth global record of a frog-parasitic species in this genus. Phylogenetic analysis based on partial SSU rDNA sequences unequivocally placed the new species within a well-supported monophyletic clade of anuran-parasitic Sphaerospora, comprising taxa distributed across the Palaearctic (Asia and Europe), Afrotropical, Nearctic, and Neotropical realms.
Heat stress is a major systemic challenge in poultry, but the role of circulating extracellular vesicles (EVs) in liver-directed adaptation remains unclear. This study investigated whether plasma-derived EVs from heat-stressed chickens (HS_EV) mediate hepatoprotective responses under thermal stress. EVs were isolated from the plasma of control and heat-stressed chickens and characterized by morphology, size distribution, and marker expression. Their biodistribution in vivo and uptake by primary hepatocytes in vitro were also evaluated. Hepatocyte injury was induced by heat exposure, and the effects of HS_EV on proliferation, apoptosis, inflammatory cytokine production, transcriptomic reprogramming, and MYD88/NF-κB/MAPK signaling were assessed. Heat stress induced systemic injury in chickens, increased the release of plasma-derived EVs, and promoted their preferential accumulation in the liver. Whole-transcriptome analysis further showed that HS_EV enhanced glutathione metabolism and related metabolic pathways while suppressing apoptosis- and inflammation-related signaling. In primary hepatocytes, HS_EV, but not control EVs, restored proliferative capacity, reduced apoptosis, suppressed the expression and secretion of IL-1β, IL-6, and TNF-α under heat stress, and was associated with attenuation of the MYD88/NF-κB/MAPK axis. These findings suggest that circulating EVs participate in adaptive intercellular communication during heat stress and identify HS_EV as a potential endogenous mediator of hepatoprotection in chickens.
Dravet syndrome (DS) is a severe, catastrophic childhood epilepsy predominantly caused by loss-of-function mutations in the SCN1A gene, which encodes the voltage-gated sodium channel Nav1.1. In this study, we evaluated the therapeutic potential of 5-(Benzofuran-2-yl)-3-(2-chloro-4-fluorobenzyl)-1,3,4-oxadiazol-2(3H)-one (GM-90663), a novel small molecule designed to address the complex pathophysiology of DS. Using scn1lab knockout (KO) zebrafish larvae-a robust vertebrate model for DS-we demonstrated that GM-90663 significantly alleviates seizure-like behavioral movements and rescues deficit in cognitive-like functions. Whole-cell patch-clamp recordings in hippocampal slices revealed that GM-90663 modulates voltage-gated Na+ channel kinetics; specifically, it suppresses slow ramp-induced currents, thereby effectively attenuating neuronal hyperexcitability. Furthermore, neurochemical profiling indicated that GM-90663 treatment leads to a marked increase in endogenous serotonin (5-HT) levels in both wild-type and KO larvae. Molecular docking simulations and subsequent in vitro enzymatic assays confirmed that this elevation in serotonin is mediated through the potent inhibition of monoamine oxidase (MAO) activity. Collectively, our findings suggest that GM-90663 exerts its anti-seizure effects through a synergistic dual mechanism-stabilizing sodium channel conductance and elevating serotonergic activity-positioning it as a promising multi-target candidate for the treatment of DS.
Duchenne muscular dystrophy (DMD) is a fatal rare disease caused by dystrophin deficiency, with no effective clinical treatments available to date. Using mdx mice as a model, this study investigated the therapeutic efficacy and interaction of mini utrophin (a truncated utrophin) and Yes-associated protein (YAP) delivered via recombinant adeno-associated virus (rAAV). Results showed that mini utrophin was efficiently expressed in mdx mouse skeletal muscle, significantly increased phosphorylated YAP (p-YAP) levels, restored the expression of dystrophin-glycoprotein complex (DGC) components (α/γ-sarcoglycans), reduced serum creatine kinase (CK) leakage, alleviated pathological damages such as central nucleation and inflammatory infiltration, and comprehensively improved grip strength, treadmill endurance, and pole climbing ability in mice. However, the co-overexpression of YAP completely antagonized these therapeutic effects, resulting in no improvement in pathological phenotypes or motor function of mdx mice. This study confirms that mini utrophin can effectively reverse DMD-related phenotypes, while excessive YAP activation abrogates its therapeutic efficacy, suggesting that precise regulation of YAP activity is required in DMD treatment and providing experimental basis for optimizing gene therapy strategies.
Staphylococcus aureus is a high-priority pathogen causing skin and soft tissue infections (SSTIs). The frequent resistance to anti-staphylococcal agents exhibited by this underscores the need for accurate diagnostics to guide effective therapy. Therefore, this study aimed to compare phenotypic and genotypic resistance in S. aureus isolates from nasal carriers and SSTIs and to elucidate gene-silencing mechanisms. In total, 355 S. aureus isolates (256 isolated from carriers and 79 from SSTIs) were studied for their phenotypic and genotypic resistance to β-lactams, macrolides, tetracyclines, aminoglycosides, and mupirocin. The silenced mupA gene (low prevalence: 0.6%; 2/335), linked to mupirocin resistance, was sequenced, and expression was assessed via reverse transcription qualitative PCR (RT-qPCR) in all mupA-positive isolates. SSTI isolates showed significantly higher resistance to erythromycin, gentamicin, and mupirocin, along with a higher prevalence of multidrug-resistant strains and ermC and tetM genes. Sequencing revealed multiple mutations in silent mupA, including a critical frameshift (c.372 delA) in a poly(A) tract that brings about premature truncation. RT-qPCR indicated upregulation of silent mupA variants and high variability in functional strains, suggesting that frameshift alone prevents resistance. These findings highlight silent resistance genes as key targets for advancing S. aureus surveillance and for combating emerging threats.