Introduction RNA viruses rapidly evolve to evade host immunity, posing a significant challenge to conventional vaccines. Natural infection, however, provides a blueprint for broad immunity by training the immune system against a diverse antigenic landscape. This study addresses the problem of creating a vaccine that mimics this complex training process. Objectives: We aimed to develop and validate a novel vaccination paradigm, “Proactive Immune Training” (PIT), designed to enhance cross-reactive neutralization breadth against variants within the predicted evolutionary space. Methods Using SARS-CoV-2 as a model, we constructed a 600-mutant mRNA library (m-library) based on 30 predicted high-frequency mutation sites. This library was encapsulated in proprietary lipid nanoparticles (LNPs) to form the PIT vaccine. We then evaluated a sequential prime-boost regimen consisting of a single-antigen (SA) vaccine followed by a PIT boost (SA + PIT) in BALB/c mice, assessing humoral and cellular immunity, and performing deep sequencing of B-cell and T-cell receptor repertoires. Results The SA + PIT regimen induced significantly broader and more potent neutralizing antibodies against a range of SARS-CoV-2 variants compared to conventional regimens. This enhanced protection was mechanistically linked to a profound diversification of the T-cell and B-cell receptor repertoires, a hallmark of superior immune adaptability. Conclusion The PIT strategy successfully guides the immune system to generate a broad and adaptive immunological response. This proactive, training-based paradigm offers a powerful and versatile framework for developing next-generation vaccines against highly variable pathogens.
INTRODUCTION:Antimicrobial resistance (AMR) poses an escalating global health threat, yet the antibiotic discovery pipeline has stagnated over recent decades. Multi-mechanistic antibacterial action may offer a promising strategy to address this challenge. OBJECTIVES:This study aimed to use clofoctol, a clinically used antibacterial agent, as the starting scaffold for cationic amphiphilic modification to generate derivatives with improved aqueous solubility, broadened antibacterial spectrum, and enhanced selectivity, followed by mechanistic characterization of the lead compound. METHODS:Minimum inhibitory concentration (MIC) and hemolysis assays were performed to assess structure-activity relationship of clofoctol derivatives. The lead compound was then systematically evaluated for bactericidal kinetics, resistance development, antibiofilm activity, salt tolerance, mammalian cytotoxicity, and therapeutic efficacy in a murine keratitis model. To investigate the mechanisms, we evaluated membrane perturbation, topoisomerase IV-mediated DNA decatenation, intracellular reactive oxygen species (ROS) and adenosine triphosphate (ATP) quantification, transcriptomic analysis, and RT-qPCR validation. RESULTS:30 was identified as the lead candidate, displaying improved apparent aqueous solubility, broadened activity against Gram-positive and Gram-negative bacteria (MICs = 0.39-3.125 µg/mL), favorable selectivity index and therapeutic index (TI = 272.5) relative to clofoctol. Furthermore, 30 exhibited rapid bactericidal kinetics, good salt tolerance, and low propensity for resistance development. In a murine keratitis model, 30 reduced bacterial burden by 4.65 and 3.81 log CFU in S. aureus ATCC29213- and P. aeruginosa ATCC9027- induced infections, respectively. These reductions were comparable to those achieved by vancomycin and gatifloxacin under the same conditions, and no observable ocular toxicity was detected. Mechanistic investigation supported membrane perturbation as the best-supported antibacterial component. Compound 30 exposure was also associated with intracellular ATP depletion, ROS accumulation, and in vitro inhibition of topoisomerase IV-mediated DNA decatenation. CONCLUSION:These results identify 30 as a promising antimicrobial agent with broadened antibacterial activity, favorable preliminary safety profiles, and multiple contributing antibacterial mechanisms.
Introduction Astrocytic-vascular crosstalk contributes to vascular repair and remodeling after brain injury. 3‑Phosphoglycerate dehydrogenase (PHGDH), the rate-limiting serine synthetic enzyme, has emerged as a crucial factor in cell growth and metabolism. However, the role of PHGDH in ischemic stroke is poorly understood. Objectives This study aimed to determine whether PHGDH-derived serine contributes to neovascularization and functional recovery after ischemic stroke. Methods We employed astrocyte-specific PHGDH deletion and PHGDH overexpressing mice to evaluate the role of PHGDH in poststroke neovascularization, vascular remodeling and functional recovery. To explore the interaction between vascular solute carrier family 38 member 2 (SLC38A2) and astrocytic PHGDH, we knocked down SLC38A2 in PHGDH-overexpressing mice using an AAV-BR1 carrying shRNA (shSlc38a2) virus. Moreover, we treated mice with mammalian target of rapamycin complex1 (mTORC1) inhibitor rapamycin to assess the effect of mTORC1 on angiogenesis and vascular integrity after ischemic stroke. Results The expression of PHGDH was significantly upregulated in astrocytes during stroke recovery, which led to the accumulation of serine in the ischemic cortex. Deletion of PHGDH in astrocytes decreased the amount of serine and enhanced vascular permeability, impaired neovascularization and the formation of functional vessels, and decreased vascular perfusion and cerebral blood flow at 14 days after ischemic stroke. In contrast, overexpression of astrocytic PHGDH increased serine levels, promoted neovascularization and vascular remodeling, and improved blood flow and long-term functional recovery. These effects were mediated through the SLC38A2 transporter, and endothelial silencing of SLC38A2 resulted in decreased vascular serine levels and the inactivation of mTORC1, and blocked neovascularization and vascular repair in PHGDH-overexpressing mice. Moreover, infusion of rapamycin impaired vascular repair and plasticity. Conclusion Our observations demonstrate that poststroke neovascularization is regulated by astrocytic-vascular interaction involving PHGDH-mediated elevation of serine.
Background Liver diseases are a major cause of illness and death worldwide. Oxidative stress is a pivotal driver in the pathogenesis of a spectrum of liver diseases, including alcoholic liver disease (ALD), metabolic dysfunction-associated fatty liver disease (MAFLD), drug-induced liver injury (DILI), and hepatocellular carcinoma (HCC). The transcription factor Nrf2, a master regulator of cellular antioxidant responses, plays a central yet context-dependent role in modulating this injury. Additionally, the gut-liver axis is a critical regulator of hepatic homeostasis. Aim of Review This review presents recent advances to propose a refined gut-microbiota-Nrf2 axis as a key mechanistic link in the treatment of liver injury. We detail how specific gut-derived microbial metabolites, such as short-chain fatty acids (SCFAs), tryptophan derivatives, and urolithins, directly or indirectly activate the hepatic Keap1/Nrf2 signaling pathway. This activation orchestrates a cytoprotective program that enhances the redox balance, promotes detoxification, and induces selective autophagy, thereby protecting against oxidative liver injury. Conversely, we examine the dual role of Nrf2, highlighting how its dysregulated and constitutive activation in established HCC can paradoxically promote tumor progression and ferroptosis resistance. Finally, we evaluate the therapeutic potential of targeting this axis using microbiome-modulating strategies, including probiotic and prebiotic supplementation, fecal microbiota transplantation (FMT), dietary intervention, and synergy with Nrf2-targeting drugs. Key scientific concepts of review This review provides an integrated framework that connects gut microbial ecology with host redox signaling, offering novel mechanistic insights and translational perspectives for the prevention and treatment of oxidative liver diseases.
BACKGROUND AND PURPOSE:Current blood-based diagnostic tests for traumatic brain injury (TBI), such as those measuring UCH-L1 and GFAP, require venous whole blood or plasma samples collection and cold-chain handling. No FDA-cleared TBI test currently supports capillary blood or dried sample formats. We propose incorporating a novel dried plasma collection device into future TBI diagnostic workflows. This format is compatible with fingerstick sampling, simplifies storage and transport, reduces biosampling burdens, and preserves key TBI biomarker detectability. Our study evaluates this device's analytical performance, focusing on biomarker stability under various storage conditions and its compatibility with multiplex TBI biomarker assays. METHODS:Capillary blood (50-70 µL) or archived plasma was applied to a Dried Plasma Biosampling Lateral Flow (DPB-LF) device. The device separates plasma via a cell filter onto a nitrocellulose strip. After air-drying for 30 min, samples were stored with desiccants under various temperatures and durations. Later, dried plasma was rehydrated and analyzed using the Quanterix Neurology 4-Plex B assay, and biomarker levels were compared to matched wet plasma samples. RESULTS:With capillary blood from healthy controls, the DPB-LF device demonstrated consistent plasma separation. SDS-PAGE analysis showed similar protein profiles in dried versus wet plasma. As proof of principle, pooled plasma samples from TBI patients and healthy controls were applied to DPB-LF devices and stored at different temperatures (4 °C, room temperature, and 40 °C) and durations (1, 7, 14, and 21 days). Upon rehydration, dried TBI plasma samples consistently yield robust recovery of GFAP, UCH-L1, NfL, and Tau signals at 4 °C across all time points and at room temperature for up to 7 days. Extended storage durations and elevated temperatures (14-21 days at room temperature, or 7-21 days at 40 °C) led to partial reductions in biomarker signal recovery, as expected. Additionally, archived plasma samples from 16 geriatric TBI patients (GCS 13-15) were applied to the DPB-LF device. Upon rehydration, biomarker levels from the dried samples showed strong correlations with matched wet plasma, with R2 ranging from 0.829 to 0.995 for GFAP, NfL, Tau, and UCH-L1. We further examined the DPB-LF device using plasma samples from a second independent cohort of 44 adult TBI subjects (GCS 3-15) collected on post-injury day 1. Dried plasma demonstrated robust prediction of cranial lesions on computed tomography, with AUC/ROC values of 0.756, 0.734, 0.738, and 0.703 for GFAP, NfL, Tau, and UCH-L1, respectively. All four biomarker levels measured in dried plasma also showed significant correlations with injury severity categories (GCS 3-8, 9-12, and 13-15) using nonparametric Kruskal-Wallis testing. CONCLUSION:The DPB-LF device enables minimally invasive capillary blood collection and stable dried plasma storage, supporting multiplex TBI biomarker analysis. This technology shows promise for clinical and field use, with potential application in future TBI studies and diagnostic purposes.
Introduction Per- and polyfluoroalkyl substances (PFASs) have been implicated in promoting the progression of intestinal inflammation. However, the specific mechanisms remain unclear. Objectives This study aimed to elucidate how PFASs exacerbate inflammatory bowel disease (IBD) progression by directly influencing macrophages, investigate the underlying molecular mechanisms, and explore potential interventions. Methods Primary mouse peritoneal macrophages (PMs) and a chronic colitis model were utilized to investigate the impacts of three typical PFASs, perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), and perfluorodecanoic acid (PFDA), on the progression of IBD to clarify the underlying mechanisms and potential interventions. Results Our study found that PFNA induced significant proinflammatory responses, characterized by marked upregulation of IL-1β and IL-18 under environmentally relevant concentrations. Mechanistically, PFNA activated the caspase-11-dependent noncanonical NLRP3 inflammasome, thereby inducing dose-dependent upregulation of caspase-1, N-GSDMD, IL-1β, and IL-18. Both caspase-11 knockdown and Z-VAD-FMK inhibition attenuated these inflammatory effects. Molecular docking suggested that PFNA might induce NLRP3 inflammasome activation by interacting with pro-caspase-11. In vivo, PFNA also aggravated IBD progression through the same pathway. Conversely, based on our previous study, myricetin was found to alleviate PFNA-induced inflammation by inhibiting the caspase-11/NLRP3 axis. Conclusion Our study reveals a molecular axis through which PFNA exacerbates intestinal inflammation and identifies myricetin as a potential intervention for IBD associated with PFNA exposure. These findings provide insights into the mechanisms of PFNA-induced intestinal toxicity and may guide the development of strategies to mitigate PFNA-associated intestinal inflammation.
INTRODUCTION:Microglia-mediated neuroinflammation hallmarks the pathophysiology of depression. We recently identified 6-shogaol (6Sh) from the ginger root (Rhizoma Zingiberis Recens) as a potent anti-neuroinflammatory and antidepressant drug candidate. OBJECTIVES:This study aimed to resolve the bioavailability and cost-effectiveness of 6Sh for the development of novel anti-neuroinflammatory drugs against depression. METHODS:A biomimicry 6Sh analog (6SA) with an amide moeity is chemically synthesized by directly coupling vanillylamine with trans-2-octenoic acid and pharmacologically evaluated in a mouse model of corticosterone-induced depression. Single-cell RNA sequencing (scRNA-Seq) was performed to profile the brain spatial transcriptomic responses to 6SA for molecular insights to define the antidepressant mechanisms. RESULTS:As results, 6SA outperforms the parent 6Sh in terms of water solubility, oral bioavailability, cytotoxicity, cost-effectiveness and pharmacokinetic profile. Remarkably, 6SA more effectively attenuated neuroinflammation and depressive-like behaviors while demonstrated good safety profile in mice. Mechanistically, the scRNA-Seq results highlighted two important differentially expressed genes (i.e., Glul and lncRNA Gm57375) in the prefrontal cortex microglia. Strikingly, in vivo and in vitro experiments indicated that 6SA regulated Glul and Gm57375 expression in a TRPV1-dependent manner. CONCLUSION:Thus, 6SA may be a promising drug candidate with the capacity to tune neuroinflammation via targeting the TRPV1-Glul-Gm57375 axis in microglia against depression.
Introduction Gray mold is an important fungal disease caused by Botrytis cinerea which threatens global agriculture. As chemical control faces limitations, biological control using Bacillus has gained attention for its environmental friendliness and growth promotion. However, their ecological basis and application potential in mulberry gray mold control remain insufficiently understood. Objective This study aimed to evaluate the biocontrol efficacy of the mulberry derived endophytic strain Bacillus velezensis ZJU_268 and to investigate its associated effects on plant growth, root-associated microbiomes, and metabolic profiles. Methods Greenhouse assays were combined with genomic and comparative genomic analyses, amplicon sequencing, non-targeted metabolomics, and functional validation of isolated microbes and metabolites to assess the effects of ZJU_268 and its cell free supernatant (CFS) on mulberry seedlings. Results This study isolated a mulberry derived endophytic bacterium, B. velezensis ZJU_268, which exhibits strong antifungal activity and reduces the incidence of gray mold in mulberry seedlings. Whole-genome sequencing and comparative genomic analyses revealed strain-specific regions and genes associated with root colonization, stress adaptation, and antimicrobial biosynthesis. Both live cells and CFS significantly promoted seed germination, seedling growth, and biomass accumulation in a dose dependent manner. Amplicon sequencing showed that ZJU_268 and its supernatant reshaped the mulberry root microbiome, enriching beneficial bacterial and fungal taxa while reducing potentially pathogenic members. Cultivable members of the enriched microbiota displayed strong antifungal activity against B. cinerea and promoted mulberry growth. Metabolomic profiling further showed that ZJU_268 and its supernatant were associated with marked metabolic shifts in mulberry roots, accompanied by the accumulation of selected metabolites that supported the growth of representative enriched isolates. Conclusions This study demonstrates that ZJU_268 suppresses gray mold and promotes mulberry growth in association with direct antagonistic activity, microbiome restructuring, and holobiont-level metabolic shifts, providing a promising biological strategy for sustainable mulberry disease management.
INTRODUCTION:Amplification of chromosome 12q13-15 spanning MDM2 and CDK4 genes serves as a molecular diagnostic hallmark of dedifferentiated liposarcoma (DDLPS), an aggressive soft-tissue sarcoma. Epigenetic activation of master transcription factors (RUNX proteins, FOSL2, and MYC) establishes a self-reinforcing oncogenic transcriptional circuitry in DDLPS. Nevertheless, the collaborative interplay between genomic alterations and epigenetic dysregulation in defining DDLPS cell identity remains elusive. OBJECTIVES:This work aimed to elucidate the primary genetic drivers and mechanistic basis of DDLPS-specific core transcriptional regulatory circuitry. METHODS:We performed integrative chromatin profiling analysis of DDLPS clinical specimens and cell lines to map cis-regulatory landscapes. Cistromes of MDM2, JUN, and E2F1 were delineated through chromatin immunoprecipitation sequencing in two DDLPS models. Essential driver functions and transcriptional regulatory effects of key regulators were assessed via various genetic manipulation approaches. Synergistic interactions between BET-targeting agents and MDM2/p53 or CDK4 inhibitors were quantified by cell viability assays. In vivo xenograft assays evaluated the oncogenic potential of key regulators and the therapeutic efficacy of novel strategies. RESULTS:Co-amplification of MDM2, CDK4, and JUN during sarcomagenesis converges with BET protein-dependent chromatin remodeling to fuel feed-forward transcriptional circuits among master transcription factors. Mechanistically, excessively expressed MDM2 stabilizes the core regulatory circuitry by forming chromatin-bound complexes with JUN/FOSL2 at cis-regulatory elements, especially super-enhancers across DDLPS genome. Concurrently, CDK4 maintains expression of E2F1 which further fosters transcriptional output of master transcription factors in DDLPS cells. Leveraging DDLPS-selective overexpression of MDM2 and its E3 ligase activity, targeted degradation of BET proteins by MDM2-recruiting proteolysis targeting chimera selectively disrupted the core regulatory circuitry, suppressing DDLPS growth and exhibiting strong synergy with CDK4 inhibitor. CONCLUSION:DDLPS-associated genomic lesions collaborate with BET-dependent chromatin regulation to establish disease-sustaining transcriptional circuitry. Our findings also provide a mechanistic rationale for harnessing MDM2's E3 ligase activity to therapeutically degrade oncoproteins in MDM2-amplified malignancies.
Background “Hidden hunger” caused by zinc (Zn), iron (Fe), and selenium (Se) deficiencies remains a major global health challenge, especially for populations relying on cereal-based diets in developing countries. While conventional food fortification and dietary supplementation work, biofortification of staple cereals (wheat, maize, and rice) through agronomic practices and breeding/biotechnological approaches offers a sustainable, food-based solution with long-term impact. However, existing literature has two key gaps. (1) Fragmented analysis: a lack of systematic cross-nutrient, cross-crop analysis of Zn/Fe/Se biofortification feasibility across wheat, maize, and rice. (2) Disconnection from industry: insufficient integration of agronomic and breeding/biotechnological technologies with food processing, market demand, and consumer health outcomes—failing to scale up trial results into industrialized products. Aim of Review This review seeks to address the aforementioned knowledge gaps and outline a conceptual framework to help bridge the “trial-to-industry” divide in Zn/Fe/Se biofortification of staple cereals, aiming to support researchers, policymakers, and food enterprises in joint efforts to mitigate global micronutrient malnutrition and advance the United Nations 2030 “Zero Hunger” Goal. Key scientific concepts of review The biofortification feasibility (levels of difficulty, similarities, and differences) of Zn, Fe, and Se in wheat, maize, and rice was systematically compared. It is easier to biofortify Se than Zn and Fe. Recent agronomic innovations, including drone-based foliar spraying, nanofertilizers, and integrated soil-crop system management, show significant promise for scalable biofortification. To develop consumer-acceptable micronutrient-enriched functional agri-food products complying with nutrition labeling and health claim regulations, several critical advances are needed: improved translocation and biosynthesis of Zn/Fe-nitrogen/protein complexes into the endosperm, advanced Zn/Fe-preserving “whole-ingredient” processing and selenomethionine extraction, and strengthened clinical evidence, policy frameworks and market guidance (particularly for Se safety). Finally, this review outlines a staged, comprehensive roadmap for integrating innovation across the industrial chain and clarifies the industrialization progress and technical maturity of each micronutrient.
INTRODUCTION:Long noncoding RNAs (lncRNAs) are key regulators of vascular endothelial function. The lncRNA differentiation antagonizing non-protein coding RNA (DANCR) is implicated in cell proliferation and inflammatory responses; however, its specific role in atherosclerosis remains undefined. OBJECTIVE:To investigate the role of DANCR in modulating endothelial adhesive capacity and atherosclerotic plaque instability. METHODS:DANCR expression was profiled in vascular tissues and cell lines using RNA fluorescence in situ hybridization and real-time qPCR. Endothelial-specific DANCR-knockout mice were generated and injected with recombinant adeno-associated virus carrying murine PCSK9 to induce atherosclerosis. Chromatin isolation by RNA purification followed by sequencing was performed to identify potential targets of DANCR. Plasma DANCR level was measured in healthy subjects (n = 42) and in patients with mixed plaques detected by coronary computed tomography angiography (n = 30). RESULTS:DANCR expression was predominantly expressed in endothelial cells and was significantly lower by 43% in the endothelium of human carotid plaques than normal vessels. Endothelial-specific knockout of DANCR in mice led to a 78% increase in aortic plaque area and a 1.1-fold elevation in the plaque instability index, characterized by enlarged necrotic cores, elevated type III/I collagen ratio, and increased macrophage infiltration. The ribosomal protein L22 (RPL22) was identified to be a target of DANCR which repressed its transcriptional expression. Endothelial-specific knockdown of RPL22 reversed the plaque progression and instability induced by DANCR deficiency in vivo. In vitro, DANCR reduced endothelial adhesion capacity and the expression of ICAM1 and VCAM1 via inhibition of the RPL22/p53 pathway. Clinically, plasma DANCR level was lower in patients with mixed plaques compared with control subjects. CONCLUSIONS:Endothelial DANCR deficiency promotes atherosclerotic plaque instability through activation of the RPL22/p53 pathway, suggesting DANCR as a potential protective factor and therapeutic target in atherosclerosis.
Introduction Liquid-liquid phase separation (LLPS) drives cellular organization through scaffold proteins that initiate condensate formation and client proteins that partition into them. Current computational tools lack precision in differentiating these functional classes and identifying key intrinsically disordered regions (IDRs), limiting mechanistic understanding of LLPS in health and disease. Objectives This study develops PhaseOM, a unified computational framework to: (1) differentiate scaffold versus client proteins; (2) identify LLPS-associated IDRs; and (3) pinpoint critical functional residues, enabling systematic analysis of LLPS determinants. Methods We constructed a multi-task predictive framework. Standard datasets were curated from drLLPS and PDB. PhaseOM integrates four specialized models: (1) a Graph Attention Network fusing ProtT5 sequence embeddings and ESMFold-predicted structural features for scaffold identification; (2) an ensemble classifier leveraging 22 optimized physicochemical and structural attributes for client discrimination; (3) a model based on 17 disorder-related features for IDR detection; and (4) a deep multi-layer perceptron with multi-head self-attention for key residue prediction. The performance of these models was thoroughly assessed using independent test datasets to ensure robustness and reliability. Results PhaseOM outperformed Seq2Phase on independent tests, achieving scaffold AUC of 0.9954 and client AUC of 0.9474 (MCC 0.7514). It improved client AUC by 0.21 and scaffold AUC by >0.07 over baseline. Key findings reveal that scaffolds are enriched in tyrosine/arginine, facilitating multivalency, while clients exhibit expanded conformations with cysteine/histidine enrichment. Empirical validation on α-synuclein isoforms confirmed high-fidelity predictions, with residue-level IDR accuracy ranging from 76% to 91%. PhaseOM provides the first unified platform for systematic proteome-wide phase separation analysis. Conclusion PhaseOM provides the first unified platform for comprehensive LLPS analysis, revealing distinct mechanistic principles and enabling rational design of biomolecules with tunable phase behavior for therapeutic applications.
Introduction Orthodontic tooth movement (OTM) relies on mechanical force that triggers inflammatory responses in periodontal tissue, orchestrating alveolar bone remodeling. Potassium (K+) efflux plays a key role in inflammation and tissue remodeling. The calcium-activated potassium channel KCNN4 regulates immune responses, but its role in inflammatory signaling in periodontal ligament cells (PDLCs) under mechanical stress remains unclear. Objectives This study aimed to investigate how mechanical force activates KCNN4 and regulates downstream inflammatory signaling in PDLCs. Methods Human PDLCs (hPDLCs) were subjected to compressive force in vitro. KCNN4, FOXO1, and USP46 were interfered with using pharmacological inhibition (senicapoc) or siRNA-mediated knockdown. Potassium and calcium fluxes, NLRP3 inflammasome activation, and downstream cytokine expression were assessed via fluorescence imaging, western blotting, qPCR, and immunoprecipitation. Chromatin immunoprecipitation and RNA-seq analyses were performed to identify transcriptional regulators of USP46. Conditioned medium from hPDLCs was applied to THP-1-derived macrophages to evaluate macrophage inflammatory polarization. In vivo, a rat OTM model was used to examine alveolar bone remodeling and local inflammation under KCNN4 inhibition. TRAP staining, immunohistochemistry, and immunofluorescence to evaluate osteoclast activity and NLRP3-mediated inflammatory signaling in the periodontal ligament. Results Mechanical force induced KCNN4-mediated K+ efflux in hPDLCs, promoting FOXO1 dephosphorylation and nuclear translocation. Activated FOXO1 transcriptionally upregulated the deubiquitinase USP46, which in turn deubiquitinated NLRP3, leading to inflammasome activation. Inhibition of KCNN4, FOXO1, or USP46 attenuated NLRP3 activation and downstream IL-1β maturation. Conditioned medium experiments demonstrated that this signaling cascade promoted macrophage inflammatory activation, while senicapoc reversed these effects. In vivo, KCNN4 inhibition suppressed local NLRP3-mediated inflammation, reduced osteoclast differentiation on the pressure side, and attenuated OTM. Conclusions Our findings reveal a mechanotransduction axis in PDLCs whereby mechanical force activates KCNN4, driving FOXO1-dependent USP46 transcription and NLRP3 inflammasome activation. This signaling cascade translates transient mechanical stimuli into an inflammatory cascade, triggering macrophage-mediated alveolar bone remodeling during OTM. Importantly, KCNN4 is required for proper inflammatory responses and OTM, highlighting its potential role in orthodontic therapy
INTRODUCTION:Intramuscular fat, known as "snowflake" pattern in pork, critically determines meat tenderness, juiciness, flavor, and overall consumer preference. Cultured meat has emerged as a promising solution to address future protein demands sustainably. However, producing cultured meat with authentic marbling remains challenging, as myocytes and adipocytes require distinct microenvironments for differentiation. OBJECTIVE:This study aimed to develop a co-differentiation strategy independent of conventional hormonal adipogenic cocktails (IBMX and dexamethasone) to simultaneously induce myogenesis and adipogenesis in a single culture system, thereby enabling the production of structured cultured meat with microscale muscle-fat patterning. METHODS:An edible κ-carrageenan-konjac glucomannan (CK) hydrogel was developed as a versatile platform for cultured meat. A 3D co-culture system was established by encapsulating porcine muscle satellite cells (pMuSCs) and fibro-adipogenic progenitors (pFAPs) within the CK scaffold, followed by differentiation induction using an eicosapentaenoic acid (EPA, C20:5n-3)-based strategy. The physicochemical properties of the resulting cultured meat induced with EPA (CM-EPA) were rigorously characterized using texture profile analysis and nutritional composition analysis, with comparisons made against horse serum-induced controls (CM-HS) and commercial streaky pork. RESULTS:The CK hydrogel exhibited tunable, biomimetic mechanical properties and a porous microstructure that supported the proliferation and differentiation of both cell lineages. Notably, the EPA-based induction strategy significantly enhanced lipid accumulation while simultaneously supporting myotube formation from pMuSCs, effectively mitigating the myogenesis inhibition associated with conventional hormonal adipogenic cocktails. The co-cultured meat exhibited significantly improved textural properties, such as hardness and chewiness, which closely resembled those of real streaky pork. Its nutritional profile also showed elevated protein and fat content relative to the CM-HS control. CONCLUSION:This work establishes a versatile scaffold-based platform for producing structured cultured meat with integrated muscle and fat components, offering a promising strategy to replicate the textural and nutritional qualities of traditional meat.
BACKGROUND:Banana, the fourth most important food crop globally, is severely threatened by Panama disease caused by Fusarium oxysporum f. sp. cubense (Foc). The continued spread of Foc tropical race 4 (TR4), with recent first reports in Peru and Venezuela, demands renewed attention and a critical reevaluation of eco-friendly management strategies. Biocontrol has emerged as a promising approach, but its efficacy varies widely across studies and geographic regions, and a comprehensive synthesis of research trends and outcomes is lacking. AIM OF REVIEW:This review critically synthesizes the past decade of biocontrol research on Panama disease, examining geographic and taxonomic trends, biocontrol efficiency, and methodological factors including strain isolation sources, pathogen races, plant growth media, soil amendments, and application methods. We aim to identify persistent patterns alongside novel developments, critically evaluate outcomes, discuss reasons for inconsistencies, and provide future recommendations to achieve the greatest success in combating Panama disease. KEY SCIENTIFIC CONCEPTS OF REVIEW:A systematic literature search yielded 123 studies, which were analyzed using random-effects meta-analysis with logit transformation. Research trends vary significantly by country and microbial group, revealing distinct regional and taxonomic methodological approaches and success rates. Overall, Streptomyces and Trichoderma emerged as the most effective genera, particularly against Foc TR4, with optimal conditions identified as: sterilized medium or field conditions, without amendments, multiple-strain SynComs, non-banana bulk soil isolates, multiple applications, and soil drenching or dual-application methods. However, current research remains geographically centralized and taxonomically narrow. Efficacy is highly context-dependent, shaped by geography, microbial genus, and methodological factors. The review identifies critical knowledge gaps, including limited field validation, incomplete reporting of experimental parameters, and publication bias, and calls for standardized reporting, expanded exploration of understudied regions and microbial sources, and integrated, multi-disciplinary approaches to translate research into practical solutions for sustainable banana production.
INTRODUCTION:Hormesis, characterized by low-dose stimulation and high-dose inhibition, is a biphasic regulatory phenomenon, and its underlying mechanisms remain elusive. Drug-induced liver injury (DILI) can progress to liver fibrosis, liver failure, and ultimately death, and natural products hold considerable promise for the treatment of DILI. OBJECTIVES:To identify the active constituents and underlying biphasic regulatory mechanism of Chrysanthemum indicum against DILI. METHODS:Structural elucidation of the new compounds was achieved through integrated interpretation of HRESIMS, 1D and 2D NMR, and ECD. Signaling pathway was determined by mitochondrial transplantation in vitro and in vivo and RNA sequencing. Target proteins were validated by the drug affinity responsive target stability-mass spectrometry analyses, isothermal titration calorimetry, cellular thermal shift assay, shRNA, and liver-specific knockdown mice. Protein sites were validated by truncation experiments, molecular dynamics simulations, and point mutations. RESULTS:A total of 21 guaianolide sesquiterpenoids, including 13 new ones, were isolated and identified from the flowers of C. indicum. Interestingly, the new compound chrysanthemolide I (CI) alleviated acetaminophen-induced liver injury in vitro and in vivo. Mitochondria isolated from CI-treated hepatocytes attenuated DILI. CI attenuated AMP-activated protein kinase (AMPK)-mediated mitochondrial oxidative stress while enhancing AMPK-dependent mitochondrial biogenesis and mitophagy. At low doses, CI binds directly to ALA-205 and ARG-301 of serine/threonine kinase 11 (STK11) with high affinity to activate AMPK; at medium doses, binding of CI to STK11 reaches saturation, leading to peak AMPK activity; at high doses, CI additionally binds to SER-261 of serine/threonine-protein phosphatase 2A catalytic subunit α isoform (PP2Acα) with low affinity to inhibit AMPK activation. Furthermore, liver-specific knockdown of both STK11 and PP2Acα largely diminished the protective effect of CI against DILI. CONCLUSION:Novel guaianolide sesquiterpenoid CI was identified as an affinity-dependent dual-target regulator of STK11 and PP2Acα to alleviate DILI.
BACKGROUND:The growing global emphasis on health consciousness has significantly increased demand for traditional and complementary medicine products, including functional foods, nutraceuicals, and medicine and food homologous substances (MFHs). The rising prevalence of chronic diseases and the shift toward preventive healthcare have accelerated this market expansion. Although MFHs have been applied for hundreds of years in traditional healthcare systems, contemporary research faces a critical knowledge gap in the systematic organization of scientific evidence. AIM OF THE REVIEW:This review systematically summarizes the major bioactive compounds and extraction methods of MFHs with therapeutic potential against MetS. It further discusses the molecular mechanisms underlying their effects on hypertension, hyperglycemia, dyslipidemia, and hyperuricemia, while highlighting current challenges related to safety and clinical translation. KEY SCIENTIFIC CONCEPTS OF THE REVIEW:MFHs contain diverse bioactive constituents, including polysaccharides, saponins, flavonoids, polyphenols, alkaloids, and terpenoids, which exert beneficial effects on multiple components of MetS through the regulation of oxidative stress, inflammation, energy metabolism, and other signaling pathways. Conventional and emerging extraction technologies have improved the recovery and utilization of these compounds, supporting their applications in functional foods, nutraceuticals, and therapeutic products. Despite promising preclinical evidence, further studies are required to standardize extraction processes, validate efficacy and safety, and strengthen clinical evidence to facilitate the development and application of MFHs in MetS prevention and management.
INTRODUCTION:The development of small-diameter vascular grafts remains clinically challenging. Decellularized small intestinal submucosa (SIS) is a promising candidate biomaterial for vascular grafts owing to the satisfactory biocompatibility and low immunogenicity. Nevertheless, its efficacy remains limited, primarily by thrombosis or dilation-induced failure. OBJECTIVES:This study presents a novel SIS / Poly (L-Lactide-co-caprolactone) (PLCL) composite graft co-functionalized with heparin and human adipose-derived stem cell exosomes (HASCs-Exo) , aiming to meet the standards of blood vessel replacement. METHODS:In this study, we fabricated SIS/PLCL hierarchical fibrous grafts via electrospinning, followed by a dual-functionalization strategy to load heparin and HASCs-Exo. The grafts were then evaluated in vitro and in vivo. RESULTS:Comprehensive in vitro characterization revealed that the composite graft possessed optimal surface topography, enhanced tensile strength, sustained in situ release of HASCs-Exo, and favorable hemocompatibility. In a rat subcutaneous implantation model, the HASCs-Exo-modified SIS/PLCL grafts exhibited significantly improved biocompatibility, as evidenced by reduced inflammatory cell infiltration compared to unmodified SIS/PLCL grafts. In rabbit carotid artery replacement experiments, HASCs-Exo-modified SIS/PLCL grafts demonstrated superior performance, evidenced by a higher patency rate on Doppler ultrasound and enhanced endothelialization confirmed by CD31 and eNOS immunofluorescent staining. Mass spectrometry and Western blot showed that ATP2B1 was enriched in HASCs-Exo. Mechanistic studies further demonstrated that HASCs-Exo promoted endothelial cell functionality by upregulating eNOS and VEGF through ATP2B1. CONCLUSION:This study presents a novel strategy that combinines HASCs-Exo with SIS/PLCL grafts to fabricate functional small-diameter vascular grafts, achieving dual optimization of mechanical integrity and endothelialization capacity.
INTRODUCTION:The combination of polydatin and hawthorn flavonoids (PH), a traditional Chinese medicine formulation for activating blood circulation and detoxification, has shown the potential to counteract atherosclerosis, but the mechanisms underlying its effects remain unclear. PURPOSE:To investigate the protective effects of pH on atherosclerosis and identify the key targets underlying mitochondrial homeostasis. METHODS:An ApoE-/- mouse model fed a high-fat diet (HFD) was established to evaluate the effect of pH on aortic plaques, and an oxidized low-density lipoprotein (ox-LDL)-induced human umbilical vein endothelial cell (HUVEC) injury model was established. Dynamin-related protein 1 (DRP1) knockdown, YTHDF2 knockdown, and overexpression models were used to identify the key targets. RESULTS:PH exerted potent dose-dependent anti-atherosclerotic effects in high-fat diet-challenged ApoE-/-mice, reducing atherosclerotic lesion burden by 44.8%, 49.29%, and 72.99% at low, medium, and high doses, respectively. In vivo, PH ameliorated systemic dyslipidemia by lowering circulating total cholesterol, triglycerides, low-density lipoprotein cholesterol, and very-low-density lipoprotein levels, while robustly suppressing proinflammatory cytokine expression. PH also rescued aortic mitochondrial damage, mitigated mitochondrial fragmentation, and restored mitochondrial structural integrity, as evidenced by increased mitochondrial length, improved aspect ratio, and elevated mtDNA abundance. In ox-LDL-injured human umbilical vein endothelial cells (HUVECs), PH rescued endothelial mitochondrial dysfunction and restored normal mitochondrial architecture by reversing the ox-LDL-induced declines in mitochondrial matrix diameter and aspect ratio. Mechanistically, PH inhibited DRP1 expression and Ser616 phosphorylation, blocked mitochondrial translocation of phosphorylated DRP1, and thereby preserved mitochondrial membrane potential and mtDNA content. DRP1 knockdown abrogated PH's endothelial protective functions, verifying DRP1 as a core downstream effector. PH significantly upregulated YTHDF2 expression in ox-LDL-stimulated HUVECs. Functional assays confirmed a negative YTHDF2-DRP1 regulatory axis: YTHDF2 depletion increased DRP1 and phosphorylated DRP1 levels by 31.92% and 30.79%, whereas YTHDF2 overexpression reduced their levels by 43.88% and 38.31%. Notably, YTHDF2 loss abolished PH-mediated DRP1 suppression, indicating PH alleviates DRP1-dependent mitochondrial dysfunction via a YTHDF2-dependent mechanism. CONCLUSION:PH exerts a protective effect on HUVECs and prevents AS by regulating the YTHDF2/DRP1 axis to restore mitochondrial homeostasis, thereby providing a novel therapeutic approach for AS.
