The pituitary gland is a key regulator of mammalian reproduction. By secreting the gonadotropins-follicle-stimulating hormone (FSH) and luteinizing hormone (LH)-the anterior pituitary precisely coordinates reproductive function. The hypothalamus releases gonadotropin-releasing hormone (GnRH), which in turn regulates the synthesis and secretion of gonadotropins by the anterior pituitary gland. Although GnRH-dependent regulation of pituitary gonadotropins has been extensively studied in rodents, the mechanisms by which GnRH regulates gonadotropins in the bovine anterior pituitary gland remain unclear. In this research, we primarily used primary bovine anterior pituitary cells (PCs) for the experiments, and employed the mouse gonadotropin cell line LβT2 to further validate the regulatory effect of GnRH on pituitary gonadotropin-secreting cells. We first demonstrated that GnRH promotes bovine gonadotropin synthesis and secretion and concomitantly reduces global cellular m7G methylation. Further experiments demonstrated that METTL1, a key methyltransferase responsible for m7G modification, is downregulated after GnRH treatment and functions as a negative regulator of gonadotropin synthesis and secretion. Mechanistically, experiments such as MeRIP-qPCR and RNA stability analyses showed that METTL1 mediates m7G methylation of bovine FOXO1 mRNA, thereby modulating FOXO1 stability and protein expression, and regulating gonadotropin synthesis and secretion. Collectively, these findings demonstrate that GnRH regulates gonadotropin synthesis and secretion in the bovine anterior pituitary, and reveal an important role for RNA epigenetic modification in this process.
Endometriosis (EMS) is a chronic inflammatory disease affecting 10%-15% of women of reproductive age and is a major cause of infertility. This study explores the therapeutic effects of linalool, a natural compound with antioxidant and anti-inflammatory properties, on EMS in a rat model. Twenty-four female rats were divided into four groups: sham, EMS control, and two treatment groups receiving linalool at 50 mg/kg and 100 mg/kg. EMS was induced by implanting uterine tissue, and linalool was administered for 28 days. Oxidative stress markers (MDA, SOD, GPx, CAT), inflammation indicators (TNF-α, IL-1β, NF-κB), apoptosis regulators (Bax, Bcl-2, caspase-3), autophagy marker (Beclin1), adhesion molecule (ICAM-1), and angiogenic factor (VEGF) were all assessed using biochemical and histological techniques. Linalool treatment significantly reduced lesion size and oxidative stress in a dose-dependent manner. The higher dose group (Lina 100) showed nearly normal histological features tissue histology. Linalool boosted antioxidant enzymes, reduced inflammatory signaling via NF-κB and MAPK pathways, promoted apoptosis, and suppressed angiogenesis and cellular adhesion. These findings suggest that linalool exerts multi-targeted effects to reduce EMS severity. The study highlights linalool's promise as a potential therapeutic agent for EMS, though further research in human models is needed to confirm its clinical relevance.
Antipsychotic (AP) medications, such as olanzapine, cause acute weight gain independent of hyperglycemia through a glucagon-dependent mechanism. Previous work has shown that exercise, a ketogenic diet, and fasting can protect against acute AP-induced hyperglycemia and attenuate increases in glucagon. As these interventions all increase circulating concentrations of fibroblast growth factor 21 (FGF21), we hypothesized that increasing endogenous FGF21 through treatment with fenofibrate would protect against AP-induced hyperglycemia. Male C57BL/6J mice were fed a low-fat diet with or without fenofibrate (0.2% w/w) for 1 week prior to an acute olanzapine (OLZ) challenge. Fenofibrate reduced food intake and caused weight loss while protecting against OLZ-induced increases in serum glucagon and blood glucose. Pair feeding mice with the same amount of food as fenofibrate-treated animals conferred a similar degree of protection against OLZ-induced hyperglycemia. Furthermore, an acute oral gavage with fenofibrate, despite increasing circulating FGF21 concentrations to a similar level as fenofibrate feeding, failed to protect against OLZ-induced glucose excursions. The effects of fenofibrate on weight loss and protection against OLZ-induced hyperglycemia were attenuated in FGF21-/- mice. Fenofibrate attenuated both OLZ-induced hyperlipidemia and increases in whole-body fatty acid oxidation, with the latter effects being dependent upon FGF21. Together, our findings demonstrate that fenofibrate protects against acute olanzapine-induced hyperglycemia and hyperlipidemia in a weight loss and FGF21-dependent manner.
To delineate the dynamic transcriptomic landscape of ossification of the ligamentum flavum (OLF) across normal, immature, and mature stages, identify core genes, and screen stage-specific drug candidates. Normal, immature, and mature OLF tissues (nine samples) from three patients with multilevel OLF were collected for RNA-seq. Differential expression, functional enrichment, and dynamic trend analyses were performed. Key genes were validated using an external GEO dataset. Single-cell and spatial transcriptomics were integrated to determine cellular origin and spatial localization. AI-based drug prediction and molecular docking were conducted. Between normal versus immature, 1026 DEGs were identified; between immature versus mature, 713 DEGs; and between normal versus mature, 1285 DEGs. Upregulated genes were enriched in RNA processing and ribosome biogenesis, whereas downregulated genes were enriched in ECM-receptor interaction. LTBP4 showed progressive downregulation during ossification (AUC = 0.88). Single-cell transcriptomics and spatial transcriptomic analysis of a public multi-omics dataset (GSE255942) revealed that LTBP4 is expressed in fibroblasts, with lower expression in ossified than normal tissues. Virtual knockout of Ltbp4 in fibroblasts recapitulated the OLF transcriptomic signature. AI-based drug prediction identified stage-specific candidates; BRD-K59831625 exhibited the lowest binding energy with LTBP4 (-8.9 kcal/mol). Sustained LTBP4 downregulation drives OLF progression, with fibroblasts as core effector cells. Stage-specific drug candidates were identified. These findings provide new targets for molecular diagnosis and precision treatment of OLF.
Diabetic peripheral neuropathy (DPN), a severe complication of Type 2 diabetes (T2D), is marked by progressive distal-to-proximal axonal degeneration. Although animal models have advanced our understanding of DPN pathogenesis, preclinical successes rarely translate into effective therapies, underscoring the need for models that more faithfully mirror human disease. In this study, we characterized two polygenic mouse strains, NONcNZO10/LtJ (RCS10) and TALLYHO/JngJ (TH), and found that both models developed obesity, hyperglycemia, dyslipidemia, and DPN by 24 weeks of age. However, hyperinsulinemia was observed only in RCS10 mice. Compared to monogenic and high-fat diet-induced models, the metabolic and neuropathic phenotypes of RCS10 and TH more closely resembled those of human DPN. In the second phase of the study, RCS10 mice were subjected to a calorie-restricted diet (60% of standard intake) for 8 weeks, which improved metabolic health and restored large fiber function, although intraepidermal nerve fiber density remained unchanged. Together, these findings identify RCS10 and TH mice as clinically relevant polygenic models for studying DPN pathogenesis and evaluating targeted therapeutic strategies.
In contrast to mammals, adult zebrafish (Danio rerio) undergo neuronal regeneration following spinal cord injury (SCI), thereby allowing successful functional recovery. The adaptive regenerative response to SCI in zebrafish includes injury-induced proliferation of ependymal radial glial cells (ERGs) and neurogenesis. The molecular mechanisms underlying injury-induced neurogenesis in "regenerative" species, such as zebrafish, have not been fully elucidated and are an area of great interest. In mammals, the purinergic receptor P2Y2 has been shown to mediate NSC proliferation and neurogenesis, but its potential role following SCI or in zebrafish is unknown. Here, we found that P2Y2 is dysregulated following SCI in adult zebrafish, and pharmacological inhibition decreased swim capacity 2 weeks after injury. While P2Y2 was expressed on actively proliferating ERGs, antagonism of this receptor did not affect their proliferation levels. However, P2Y2 inhibition did attenuate SCI-induced neurogenesis and is strongly expressed by immature and mature neurons in adult zebrafish. Overall, P2Y2 serves as a pro-neurogenic factor during injury-induced neurogenesis and contributes to the recovery of swimming behavior in adult zebrafish.
IgA nephropathy (IgAN) is a progressive glomerular disease marked by mesangial IgA deposition, inflammation, podocyte injury, and fibrogenesis. Although understanding of its pathophysiology has advanced, effective disease-modifying therapies are still limited. Sarsasapogenin (SAR), a natural steroidal sapogenin, demonstrates anti-inflammatory, antioxidant, and antifibrotic properties by modulating signaling pathways such as NF-κB and TGF-β. The present study investigated the renoprotective effects of SAR in an experimental model of IgAN. IgAN was induced in Sprague-Dawley rats using bovine serum albumin (BSA), carbon tetrachloride (CCl4), and lipopolysaccharide (LPS). SAR was administered at 60 mg/kg/day from week 7 through week 12. Renal function, histopathological changes (Oxford MEST-C classification), and immunohistochemical, ultrastructural, and molecular alterations were assessed. Induction of IgAN resulted in significant proteinuria, renal dysfunction, mesangial IgA deposition, and podocyte alterations, accompanied by increased expression of IL-6, TGF-β1, SMAD3, fibronectin, and α-SMA. Masson's trichrome staining did not reveal excessive collagen deposition, indicating early fibrogenic activity rather than established fibrosis. SAR treatment significantly reduced proteinuria and improved renal function, accompanied by decreased IL-6-mediated inflammation and reduced activation of the TGF-β1/SMAD3 pathway. SAR also preserved podocyte-associated proteins, including nephrin and podocin, and lowered pre-fibrotic markers. Ultrastructural analysis confirmed preservation of podocyte architecture, with reduced foot process fusion and mesangial matrix expansion. Overall, sarsasapogenin modulated multiple targets in experimental IgAN by reducing inflammation and early fibrogenesis while supporting podocyte integrity. These findings indicate that SAR is a promising therapeutic candidate for early IgAN, although further mechanistic and long-term studies are warranted.
Intracranial aneurysms (IAs) represent a significant and potentially life-threatening category of disease, and there is currently a lack of effective treatment options aimed at preventing the progression of the disease. Accordingly, this study is dedicated to exploring and identifying effective drug targets that can help in the prevention of both the formation and rupture of IAs, along with a detailed examination of the underlying potential mechanisms involved in these processes. The data related to IAs for this research was obtained from the ISGC Biobank and UK Biobank. Then, we investigated the possible biological functions and unintended consequences of targeting the specific genes that were highlighted in IAs by using mediation analysis, virtual knockout experiments, and PW-MR studies. A total of 5 unique potential drug targets for IAs (FKTN, MAP3K1, PSMA4, SLC22A4, ADAM17), 4 unique potential drug targets for SAH (PSMA4, ADAM17, GPR160, SLC22A4), and 2 unique potential drug targets for UIA (SLC22A4, PRCP) were identified across brain or blood samples. Among the various candidates identified, SLC22A4 has emerged as a promising potential drug target, showing significant expression levels in both blood and brain tissues. Additionally, phenome-wide MR of SLC22A4 across 32 selected phenotypes did not identify statistically significant adverse associations after FDR correction. Virtual knockout (KO) experiments on SLC22A4 revealed that SLC22A4 KO disrupted 81 genes, all of which are involved in IAs-related pathways. Besides, we recognized BRD-K85337334 as potential candidates for targeting SLC22A4. This research indicates that an increase in SLC22A4 gene expression within the blood or brain is directly linked to a heightened risk of IAs rupture, which will aid in prioritizing the development of drugs for IAs.
Heterotopic ossification (HO) of the Achilles tendon induces pain and impairs tendon mechanical function, yet its pathogenic mechanisms remain largely unknown. Given that neurotransmitters, inflammatory cytokines, and mechanical loading are highly associated with HO incidence, we raised a hypothesis that activation of sensory neurons within the injured Achilles tendon may be one critical cause driving HO progression. This present work demonstrates that Netrin-1, predominantly secreted by macrophages at the injured site, profoundly promotes the regeneration of Piezo2-expressing sensory nerves in the Achilles tendon of mice after puncture. This is accompanied by an aberrant rise in the pro-osteogenic neuropeptide calcitonin gene-related peptide (CGRP) in both sensory nerves at the injury site and the dorsal root ganglion (DRG), particularly under the synergistic influence of mechanical stimulation and prostaglandin E2 (PGE2), a pro-inflammatory cytokine initially produced by macrophages. Consequently, osteogenic differentiation of tendon-derived stem cells (TDSCs) is enhanced, which in turn further stimulates PGE2 secretion from osteoblasts. Of note, such an effect and subsequent HO formation can be significantly attenuated by tail suspension treatment or by blocking signaling pathways mediated by CGRP, Netrin-1, PGE2, or Piezo2. Together, these findings elucidate the crosstalk among macrophages, osteoblasts, and Piezo2+ sensory neurons in the injured Achilles tendon, highlighting the critical role of sensory hyperinnervation in promoting HO progression.
Glaucoma is the leading cause of irreversible blindness worldwide, primarily driven by the progressive loss of retinal ganglion cells (RGCs) under pathological high intraocular pressure (ph-IOP). Despite the established role of ferroptosis in RGC degeneration, specific molecular targets that can be used for clinical intervention still need to be optimized, and the slow onset of conventional gene therapy vectors is incompatible with the acute clinical course of glaucoma. Here, we integrate single-cell RNA sequencing and spatial transcriptomics to profile the dynamic transcriptomic landscape of the rat retina across acute, subacute, and chronic stages of ph-IOP injury. Through ferroptosis-focused screening of an early-activated RGC gene cluster, we identify the lipid metabolism regulator phosphatidylethanolamine-binding protein 1 (PEBP1) as a candidate mediator of RGC ferroptosis. We demonstrate that Pebp1 is specifically upregulated in injured RGCs with a trajectory mirroring ferroptosis pathway activation, and that AAV-mediated Pebp1 knockdown suppresses ferroptosis through the GPX4/ACSL4 signaling axis, thereby preserving RGC survival, retinal structure, and visual function. To overcome the critical time-window bottleneck-the several weeks delay required for AAV-mediated silencing versus the rapid, irreversible RGC loss in acute glaucoma-we engineer Exosomes-siPebp1, a mesenchymal stem cell-derived exosome system loaded with siPebp1, which enables immediate single-dose intervention post-injury. This system exhibits efficient RGC uptake, prolonged intraocular retention, and robust target gene silencing, and, in a head-to-head comparison, significantly outperforms unloaded exosomes, liposomal formulations, and AAV vectors in RGC protection, without detectable acute systemic or local toxicity. Collectively, this study implicates Pebp1 in ph-IOP-associated RGC ferroptosis and supports exosome-mediated siRNA delivery as a rapid, cell-free intervention strategy for acute glaucomatous injury.
Bile acid biology has advanced through significant conceptual shifts. Once understood primarily as biological detergents, bile acids are now recognized as signaling molecules and, more recently, as a chemically diverse set of host- and microbe-derived metabolites. The 2020 discovery of microbially conjugated bile acids (MCBAs) and the expansion of the recognized catalog from approximately 20 species to more than 200 mark a new phase in this trajectory. Clinical translation has not kept up. Direct farnesoid X receptor (FXR) agonism failed twice in trials for non-alcoholic steatohepatitis, later received a serious liver injury safety communication from the U.S. Food and Drug Administration, and was voluntarily withdrawn from the United States market. The standard of care for bile acid diarrhea and post-cholecystectomy diarrhea still relies largely on chemical binding with drugs introduced in the 1960s and 1970s. This Perspective argues that the gap between bile acid biology and bile acid medicine persists in part because the field has not been organized around the bile acid pool as a shared measurable output. Researchers studying dietary modulators of the gut microbiome have worked in separate communities around fiber, fermented foods, polyphenols, protein, and dietary fat. Each of these inputs can shape bile acid metabolism, yet many intervention studies do not measure it. The most immediately implementable bile acid-targeted strategy is specified dietary intervention designed with measurable bile acid outcomes and evaluated with the precision of pharmacological therapy. Four recommendations follow: intervention studies should routinely measure bile acid outcomes; clinicians should test bile acid metabolism in conditions involving dysregulation; regulators should develop a framework for multicomponent dietary therapies; and researchers should build infrastructure for population-scale longitudinal monitoring.
Cisplatin causes nephrotoxicity by accumulating in renal tubular epithelial cells (RTECs). Astragaloside IV (ASIV) shows renoprotective potential, but its mechanisms remain poorly understood. Cisplatin induced nephrotoxicity was established in 8-week-old male C57BL/6 mice via intraperitoneal administration of cisplatin at 20 mg/kg for 48 h. For in vitro studies, HK-2 human proximal tubular epithelial cells were exposed to 50 μM cisplatin for 24 h. Multi-omics approaches were employed to identify novel mechanisms by which ASIV ameliorates cisplatin-induced proximal tubular injury. ASIV markedly reduced serum creatinine and urea nitrogen levels in mice, and ameliorated cisplatin-induced proximal tubular injury both in vivo and in vitro. Moreover, ASIV restored mitochondrial damage, upregulated protein expression of PGC-1α, TOMM20, and PINK1 in RTECs. Mechanistically, RNA-seq and scRNA-seq revealed that cisplatin predominantly affected ADRA1A-mediated mitochondrial biogenesis and mitophagy in proximal tubular cells, accompanied by suppression of the AMPK/FOXO3A pathway. Notably, ASIV upregulated ADRA1A expression, thereby facilitating AMPK and FOXO3A phosphorylation and consequently enhancing mitochondrial biogenesis and mitophagy. Furthermore, dabuzalgron (a selective ADRA1A agonist) recapitulated the protective effects of ASIV. In contrast, the renoprotective action of ASIV against cisplatin-induced proximal tubular injury was largely abrogated by the ADRA1A antagonist tamsulosin in vivo and by ADRA1A-specific siRNA in vitro. These findings identify ASIV as a highly promising renoprotective agent that upregulates ADRA1A expression and activates the AMPK/FOXO3A axis to enhance mitochondrial biogenesis and mitophagy, thereby counteracting cisplatin-induced proximal tubular injury.
African swine fever (ASF) is an acute, febrile, and highly contagious infectious disease of swine with the etiological agent of African swine fever virus (ASFV). The mortality rate of virulent strains is as high as 100%. Strengthening biosafety is so far the most effective way to prevent and control ASF. Therefore, it is urgent to develop a safe and effective vaccine. In this study, a Genotype II live-attenuated ASF vaccine bearing 24 genes deletion in 3 independent regions was constructed based on the highly virulent Eurasian strain ASFV CN/GS 2018 backbone. The resulting mutant ASFV-Δ24 is characterized by complete deletion of 24 genes distributed in 3 genomic positions of 852 to 11 468, 19 732 to 22 929, and 179 519 to 180 617, among which MGF100 and whole MGF300 families are pioneeringly removed. The ASFV-Δ24 displayed a delayed and reduced replication kinetics as well as aberrant icosahedral empty particles devoid of a nucleoid when compared to the parental virus. Animal experiments showed that ASFV-Δ24 was completely attenuated in animals as evidenced by stable body temperature and no ASF-compatible clinical signs in vaccinated pigs. The ASFV-Δ24 could provide complete homologous protection against lethal challenge, as vaccinated pigs demonstrated boosted antibody response, transient but low levels of viremia in blood and virus titers in organs as well as almost undetectable viral shedding. Gene deletions in multiple regions are helpful for prevention of virulence reversion. These results indicate that ASFV-Δ24 can be used as an effective and promising candidate vaccine to control the spread of ASFV.
Annexin A4 (A4) is a negative modulator of adenylyl cyclase type 5 (AC5) with increased expression in failing human hearts. Here, we investigated whether A4 deficiency contributes to cardiac electrical and structural remodeling induced by chronic stimulation of β-adrenergic receptors (βAR). A4-deficient mice (gene trapped, GT) and wild-type (WT) were infused for 7 days with isoprenaline (ISO) or NaCl as control. Myocytes of ISO-treated GT (GTISO) displayed more hypertrophy, increased action potential duration, reduced K+-current Ito, preserved L-type Ca2+ current ICaL with a negative shift of voltage dependence of activation, in line with increased AC/cAMP/PKA signaling, and increased NCX1 versus WTISO. At the molecular level, mRNA levels for Kcnd3, Kcnip2, Cacna1c decreased at unchanged protein levels of Kv4.2, Kv4.3, KChIP2, α1C, independent of genotype, suggesting posttranslational modifications of the channels underlying Ito and ICaL. Chronic ISO-induced βAR desensitization and redox stress were confirmed by decreased cAMP production and lower mRNA levels of Adrb1, Adcy5/6, and Sod2, and reduced response to acute ISO, with no additional genotype-dependent effects on calcium handling or contractility. Nevertheless, GTISO cardiomyocytes retained a greater cAMP response to acute ISO, in line with AC5 disinhibition and preserved β2AR and Gαs/i, suggesting genotype-dependent differences in β-adrenergic signaling under stress. mRNA levels of Anxa4 were increased in hypertrophied WTISO vs. normal WTNaCl hearts, supporting a protective role for A4. Overall, the advanced remodeling in A4-deficient myocytes detected in response to chronic βAR stimulation proposes using A4 peptide as a therapeutic tool to prevent the progression of cardiac electrical remodeling.
The human gut mycobiome plays a critical role in host immunity, yet standardized workflows for fungal internal transcribed spacer (ITS) amplicon sequencing remain elusive. Unlike the bacterial 16S rRNA gene, where the impact of DNA versus RNA templates is well-documented, such comparative benchmarks are lacking for fungal studies. Furthermore, fungal ITS regions exhibit significant length heterogeneity, posing unique challenges for denoising algorithms that were originally designed for conserved bacterial amplicons. We performed fungal ITS2 sequencing on six fecal samples, generating 12 matched DNA and RNA-derived cDNA libraries for paired methodological benchmarking. We systematically compared three representative workflows, DADA2, Deblur, and VSEARCH, to evaluate read retention, feature recovery, and preservation of ITS2 length heterogeneity. To address dataset-size limitation, we further added an independent public fecal fungal ITS2 validation cohort from PRJNA419104, including 32 baseline CDI samples and 21 metadata-confirmed healthy controls, and reprocessed this dataset using the same comparative pipeline framework. AGI-versus-healthy comparisons in the internal cohort were treated as exploratory secondary analyses rather than disease-biomarker discovery. DADA2 best preserved fungal ITS2 length heterogeneity while avoiding the fixed-length truncation artifact observed with Deblur and the feature inflation typical of permissive OTU clustering. Paired DNA and RNA profiles showed concordant dominant community structure, with no ANCOM-detected differential abundance between templates across major taxonomic levels. Exploratory AGI-versus-healthy comparisons suggested candidate differential-abundance features, but these observations were considered hypothesis-generating and were not interpreted as validated disease biomarkers. In this small-sample paired methodological benchmark, DADA2 is the preferred workflow for fungal ITS2 amplicon analysis in this paired benchmark because it preserves variable-length sequence information. DNA sequencing provides a practical primary layer for routine dominant-community profiling, whereas RNA-based sequencing can serve as an optional complementary layer when low-abundance or activity-oriented features are of specific interest. The results presented in this article are limited by the small sample size, and larger clinical cohorts are required for disease-association or biomarker inference.
High-fat diet (HFD)-induced obesity (DIO) is preceded by disruptions in endogenous circadian rhythmicity, including lengthening of its period (tau). We previously demonstrated that housing mice under a light-dark cycle (T-cycle) oscillating at their endogenous tau of 23.7 h prevents the DIO found under the 24-h T-cycle, suggesting that DIO is also secondary to disruptions caused by entrainment to non-tau-matching T-cycles. Here, we aimed to test this hypothesis by comparing energy homeostasis of low-fat diet (LFD) and HFD-fed female mice housed under four regimes: constant darkness (DD), a T-cycle oscillating at the tau of age-matched LFD-fed mice, a non-tau-matching 24-h T-cycle, or a T-cycle shorter than tau by the age-appropriate tau-24-h deviation (∆tau). In LFD-fed mice, energy homeostasis was unaffected by photic regime. In contrast, while HFD induced DIO across all regimes, onset occurred at similar times under DD and the 24-h T-cycle but was similarly delayed by 8 weeks under both the tau-like and ∆tau T-cycles. Notably, DIO onset under DD was preceded by the prevention of tau shortening, a characteristic of LFD-fed mice. Delayed DIO was not explained by reduced energy intake; instead, it was associated with a preserved locomotor activity level, suggesting higher energy expenditure, and was followed by a stronger (compared to 24-h T-cycle) tau-shortening aftereffect. In conclusion, these findings identify HFD-induced tau lengthening as a mechanism promoting early DIO under the conventional 24-h T-cycle. Importantly, they suggest that tau-shortening pharmacological or nutritional interventions may postpone DIO in susceptible individuals and settings, even during ad-libitum HFD feeding.
African swine fever virus (ASFV) causes an incurable swine disease with nearly 100% mortality, posing a catastrophic threat to global pig production. The soft tick Ornithodoros lahorensis acts as a critical biological vector that sustains persistent ASFV replication and mediates long-distance viral transmission, yet the molecular mechanisms governing ASFV-tick interplay remain poorly understood. Here, we integrated transcriptomics and metabolomics to systematically dissect molecular changes in O. lahorensis across three infection stages: Uninfected control, early infection (7 days post-infection, dpi), and late persistent infection (21 dpi). Multi-omics integration revealed that ASFV extensively remodels tick host metabolism, predominantly activating purine/pyrimidine metabolism, lipid biosynthesis, and energy metabolism. We further characterized a conserved regulatory module consisting of 12 core genes and 8 signature metabolites that collectively support ASFV genome replication and virion assembly. Three hub metabolic genes (TK1, ATP5F1B, and IMPDH) were selected for functional validation via siRNA silencing in ticks; individual gene silencing suppressed ASFV loads by 89.2%, 91.5%, and 87.8%, respectively (p < 0.001***). This work represents the first comprehensive multi-omics investigation of ASFV infection in O. lahorensis. We identified tick-specific molecular targets to block vector-mediated ASFV spread and established a standardized multi-omics analytical pipeline for tick-virus interaction research. Our findings elucidate the mechanistic basis of long-term ASFV persistence in soft ticks and deliver novel actionable clues for developing vector-targeted ASF intervention strategies.
Diabetes mellitus frequently leads to diabetic wounds (DW), a serious complication for which current treatment options remain limited. This work examined the pro-healing effects of sesamin (Sea), a major sesame lignan, on DW and the mechanisms underlying these effects. In vitro, human umbilical vein endothelial cells (HUVECs) were cultured under high glucose (HG) conditions to mimic diabetic dysfunction. Cell viability, lipid peroxidation, Fe2+ accumulation, mitochondrial function, and expression of ferroptosis-related proteins were assessed. Sirt1 knockdown was performed to verify target specificity. In vivo, a streptozotocin-induced diabetic mouse model with full-thickness skin wounds was established. Sea was administered, and wound healing rates, reactive oxygen species (ROS) levels, GPX4 expression, and transcriptomic profiles were analyzed. In vitro, Sea dose-dependently ameliorated HG-induced dysfunction in HUVECs and suppressed ferroptosis. Mechanistically, Sea upregulates Sirt1 expression, which promotes the dissociation of Keap1/Nrf2, thereby facilitating Nrf2 nuclear translocation and upregulating anti-ferroptosis proteins expression. These effects were abrogated by Sirt1 knockdown. In vivo, transcriptomic analysis revealed ferroptosis inhibition as a key mechanism underlying Sea-mediated DW healing. Sea treatment accelerated STZ-induced DW closuring, reduced ROS levels, and upregulated GPX4 expression, effects that were diminished by Sirt1 silencing. Collectively, our findings demonstrate that Sea promotes DW healing by activating the Sirt1/Keap1/Nrf2 pathway to inhibit ferroptosis, positioning Sea as a promising therapeutic candidate for DW treatment.
Cancer immunotherapy has reshaped modern oncology by enabling the immune system to recognize and eliminate malignant cells. However, many solid tumors still respond poorly because of weak T-cell activation and an immunosuppressive tumor microenvironment. Bacterial superantigens (SAgs) represent a unique class of immunomodulatory proteins capable of overcoming these limitations through direct activation of large T-cell populations. Unlike conventional antigens, superantigens bypass classical antigen processing by simultaneously binding major histocompatibility complex class II molecules and T-cell receptor Vβ domains, triggering rapid cytokine release and extensive immune activation. Although this potent mechanism has historically been associated with severe systemic toxicity, recent advances in protein engineering and targeted delivery have renewed interest in their therapeutic potential. This review discusses the structural and immunological basis of superantigen activity and highlights emerging strategies designed to improve tumor specificity and safety, including engineered low-toxicity variants, antibody-superantigen fusion proteins, nanoparticle-based delivery systems, and tumor-targeted constructs. We further examine how superantigens reshape the tumor microenvironment and synergize with immune checkpoint blockade, adoptive cell therapies, and other T-cell-redirecting approaches. Together, these advances position engineered superantigens as promising immune-amplifying platforms with the potential to complement existing cancer immunotherapies and improve responses in poorly immunogenic tumors.
Training with low carbohydrate availability (LCA) has been proposed as an independent determinant of physiological perturbations commonly attributed to low energy availability (LEA) and to increase skeletal muscle oxidative machinery, yet the effects of LCA in isolation from LEA remain unclear. We examined whether short-term carbohydrate restriction under energy balance alters endocrine and metabolic markers associated with LEA and skeletal muscle proteomic response. In a randomized crossover design, eight trained males completed 4 days of either a low-carbohydrate high-fat diet (LOW; 12% carbohydrate, 69% fat, 19% protein) or a normal-carbohydrate diet (NORM; 62% carbohydrate, 19% fat, 19% protein), while undertaking daily cycloergometer exercise (15 kcal kg FFM-1 day-1) and maintaining energy availability at 45 kcal kg FFM-1 day-1. LOW induced a clear metabolic shift consistent with LCA, evidenced by elevated circulating free fatty acids, glycerol and β-hydroxybutyrate, in fasting conditions and fat oxidation at rest and during exercise, alongside reduced exercise glucose concentrations. Despite these responses, LOW did not alter insulin, testosterone, triiodothyronine, leptin, hepcidin, or P1NP. In contrast, β-CTX increased and IGF-1 decreased relative to NORM. Muscle glycogen concentration decreased only in LOW (40% ± 14%). Proteomic analysis identified 671 proteins; 57 differentially expressed in LOW relative to NORM were limited to fatty acid metabolism pathways and suppression of ribosomal, sarcomeric, and extracellular matrix proteins. These findings indicate that isolated LCA exerts limited endocrine disruption but may selectively compromise bone turnover and muscle anabolic response, suggesting that without acute LEA, LCA has limited influence on muscle oxidative phenotype.
