The 2025 Nobel Prize in Physiology or Medicine was awarded to Mary E. Brunkow, Fred Ramsdell and Shimon Sakaguchi in recognition of their groundbreaking contributions to unraveling the mechanisms of peripheral immune tolerance. Regulatory T cells (Treg cells), as the core components maintaining peripheral immune tolerance, exhibit high plasticity and heterogeneity. Dysregulation of Treg function is closely associated with autoimmune diseases, tumor progression, and transplant rejection. Forkhead box protein P3 (FOXP3) is a key transcription factor that controls the development and function of Tregs. This review discusses the classification of Tregs into thymic-derived Tregs (tTregs), peripherally induced Tregs (pTregs), and in vitro-induced Tregs (iTregs). It also elaborates on how Treg cells exert their inhibitory functions through multiple pathways, including the secretion of inhibitory factors, metabolic interference via competitive uptake of IL-2, and direct cell-cell contact. In recent years, significant advances have been made in Treg and FOXP3 research, progressively deepening our understanding of Treg plasticity. Investigations have revealed their capacity to adapt and acquire features of effector T helper cell subsets—such as Th1, Th2, and Th17—under specific microenvironmental cues. This plasticity also poses challenges for therapeutic interventions, as Tregs can potentially lose their suppressive function and acquire pro-inflammatory properties, thereby exacerbating disease pathology. Furthermore, the concept of tissue-specific Treg specialization has emerged, highlighting distinct functional subsets resident in organs such as the gut, adipose tissue, and tumors. For instance, gut-resident Tregs maintain tolerance to commensal bacteria and dietary antigens, while tumor-infiltrating Tregs promote immune evasion by suppressing anti-tumor immunity. Concurrently, studies on the metabolic and epigenetic regulation of Tregs, including post-translational modifications of FOXP3 such as acetylation and ubiquitination, have uncovered intricate layers of control over their stability and function. Building upon these fundamental insights, this review synthesizes FOXP3-targeted therapeutic strategies. These encompass approaches to enhance Treg function in autoimmune diseases and transplantation, including adoptive cell therapies and pharmacological interventions. Conversely, strategies to antagonize Treg-mediated immunosuppression in oncology, such as immune checkpoint blockade, are discussed. Notably, the development of programmable engineered Tregs represents a particularly promising frontier for achieving antigen-specific immune modulation with enhanced precision and efficacy. However, the field of Treg research continues to grapple with several complex challenges. The deeper, underlying regulatory networks governing Treg biology remain incompletely understood. A comprehensive resolution of Treg heterogeneity is still lacking, and significant hurdles exist in maintaining the stability and function of Tregs during in vitro expansion and culture. Furthermore, the precision and efficacy of translating these findings into clinical applications require substantial improvement. Consequently, both the development of Treg-targeting pharmacological agents and the refinement of Treg-based cellular therapies demand more profound exploration. The ultimate goal is to overcome these obstacles and achieve transformative, breakthrough clinical outcomes in the foreseeable future.

Regulatory T cells (Treg cells) have reshaped modern immunology by establishing the conceptual and mechanistic foundation of peripheral immune tolerance. Since the pioneering identification of CD4⁺CD25⁺ suppressive T cells by Shimon Sakaguchi and the subsequent discovery of the lineage-defining transcription factor forkhead box P3 (Foxp3) by Mary E. Brunkow and Fred Ramsdell, Treg cells have been recognized as indispensable guardians of immune homeostasis. These advances collectively clarified that central tolerance alone is insufficient to eliminate all self-reactive lymphocytes, and peripheral tolerance—critically mediated by Treg cells—serves as a second barrier preventing pathological autoimmunity. Contemporary research has therefore expanded the functional and therapeutic significance of Treg cells across the fields of autoimmunity, cancer, transplantation, and tissue repair. Treg cells originate from two major developmental pathways: thymus-derived Treg (tTreg) cells, which arise from high-affinity self-reactive TCR interactions in the thymus, and peripheral Treg (pTreg) cells, which are induced in mucosal and other peripheral tissues via antigen stimulation under tolerogenic cytokine cues such as IL-2 and TGF-β. Their differentiation is orchestrated by a multilayered transcriptional and epigenetic network within the Foxp3 locus, including CNS0-CNS3 elements that integrate TCR, cytokine and environmental signals to support lineage stability. Treg cells are identified by a combination of surface and intracellular markers——CD25, CD127^low/-, CTLA-4, GITR, TNFR2, CD39/CD73, and Foxp3——although marker specificity varies with context, activation state, and species. Their notable heterogeneity enables Treg cells to adopt Th1-, Th2-, Th17- or Tfh-like programs through transcription factors such as T-bet, GATA3, RORγt and Bcl6, thereby permitting precise suppression of corresponding effector responses. Tissue-resident Treg subsets in adipose tissue, skin, skeletal muscle and the CNS have emerged as highly specialized regulators that integrate local metabolic and stromal signals, contributing not only to immunosuppression but also to tissue regeneration. Mechanistically, Treg cells maintain tolerance through three synergistic strategies: (1) secretion of suppressive cytokines (IL-10, TGF-β, IL-35) and cytotoxic mediators (granzyme B, perforin); (2) cell-contact-dependent interactions via CTLA-4, PD-1/PD-L1, and LAG-3 to limit dendritic cell maturation and T-cell activation; and (3) metabolic regulation including IL-2 consumption, adenosine production via CD39/CD73, cAMP transfer through gap junctions, and adaptation to hypoxic or nutrient-restricted microenvironments. Dysregulation of Treg cell quantity or function contributes directly to pathogenesis across a spectrum of diseases. In autoimmune diseases such as type 1 diabetes, systemic lupus erythematosus, rheumatoid arthritis and multiple sclerosis, impaired Foxp3 stability, epigenetic abnormalities, defective IL-2 signaling or inflammatory cytokine exposure undermine Treg suppressive capacity, facilitating excessive autoreactive T- and B-cell activation. In contrast, within the tumor microenvironment, Treg cells are often enriched through chemokine axes such as CCL22-CCR4 and reinforced by interaction with myeloid-derived suppressor cells and tumor-associated macrophages. Their enhanced metabolic fitness and suppressive phenotype enable tumors to evade immune destruction. In transplantation, Treg cells are essential for promoting graft tolerance, restraining effector T-cell activation, and facilitating tissue repair after injury. Rapid therapeutic progress has been driven by Treg-based immunomodulation. Polyclonal Treg adoptive transfer has demonstrated safety and preliminary efficacy in type 1 diabetes, autoimmune disorders, solid-organ transplantation, and graft-versus-host disease. Gene-engineered Treg therapies, including antigen-specific CAR-Treg and TCR-Treg platforms, offer superior precision and stability, enabling targeted suppression at disease sites. Additional strategies——including low-dose IL-2 therapy, small-molecule modulation, and selective depletion of intratumoral Treg using antibodies against CCR4, CCR8, CTLA-4 or CD25×TIGIT bispecifics——further expand the translational landscape. Collectively, advances in Treg biology——from lineage ontogeny and molecular regulation to specialized functions and therapeutic engineering——highlight Treg cells as central orchestrators of immune equilibrium. Continued integration of single-cell multi-omics, systems immunology and gene-editing technologies is expected to accelerate the development of highly specific, durable and safe Treg-centered therapies, ultimately enabling precision control of immune tolerance in autoimmunity, transplantation and cancer.

Regulatory T cells (Treg cells) are a specialized subset of CD4⁺ T cells defined by expression of the lineage-specifying transcription factor FOXP3 and a potent capacity to maintain peripheral immune tolerance. The modern concept of Tregs was catalyzed by Shimon Sakaguchi’s identification of CD4⁺CD25⁺ suppressive T cells and subsequent work establishing FOXP3 as a central determinant of Treg cell development and function; together with landmark FOXP3 genetic discoveries by Mary E. Brunkow and Fred Ramsdell, these advances transformed understanding of immune homeostasis and were recognized by the 2025 Nobel Prize in Physiology or Medicine. Under normal physiological conditions, FOXP3⁺ Treg cells restrain autoreactive lymphocytes, prevent excessive inflammation, and shape antigen-presenting cell activity through contact-dependent pathways and suppressive cytokines, thereby protecting tissues from immune-mediated damage. Disruption of Treg abundance, stability, or suppressive capacity can therefore lead to immune dysregulation and disease. Over the past two decades, Treg cells have become a major focus of immunology because their roles are highly context-dependent. In autoimmune and chronic inflammatory diseases, impaired Treg cell function or insufficient Treg activity contributes to loss of tolerance and persistent tissue injury, supporting therapeutic approaches designed to enhance Treg cell number, stability, and suppressive potency. In contrast, many cancers exploit Treg cells by promoting their expansion, activation, and recruitment into the tumor microenvironment (TME), where they blunt antitumor immunity by suppressing cytotoxic T-cell priming and effector function, limiting dendritic cell activation, and fostering immune escape. In both settings, immune checkpoint pathways critically influence Treg cell biology. Beyond PD-1/PD-L1 and CTLA-4, emerging checkpoints and costimulatory receptors, including TIGIT, TIM-3, LAG-3, and OX40, modulate Treg cell generation, stability, and suppressive functions, thereby shaping the balance between tolerance and immunity. Meanwhile, immunometabolic adaptations further tune Treg cell fitness and function in inflamed tissues and tumors; lipid utilization and mitochondrial programs, among other metabolic axes, enable Treg cells to persist in nutrient- and oxygen-restricted microenvironments, while microenvironmental stress can drive functional remodeling or fragility in a subset-dependent manner. In this review, we summarize the discovery and defining biological features of Treg cells, highlight core suppressive mechanisms and regulatory circuits, and synthesize evidence for the dual roles of Treg cells in preventing autoimmunity yet enabling tumor immune evasion. We further outline current and emerging therapeutic strategies aimed at augmenting Treg cell activity to restore tolerance in autoimmune disease, or selectively depleting, functionally inhibiting, and reprogramming tumor-resident Treg cells to enhance cancer immunotherapy. Overall we discuss how deeper insight into Treg heterogeneity, checkpoint control, and immunometabolic regulation may enable more precise Treg cell-directed interventions and inform next-generation immunotherapeutic combinations across immune-mediated and malignant diseases.

Metal-organic frameworks (MOFs), a class of porous crystalline materials formed by the self-assembly of metal ions/clusters and organic ligands, have shown broad application potential in the biomedical field due to their high specific surface area, precisely tunable pore structure, designable framework composition, and good biocompatibility. This paper traces the origin and development of MOFs, summarizes the contributions of the main promoters, and then systematically reviews the conventional synthesis and characterization methods of MOFs. Subsequently, it conducts an in-depth discussion around three application directions in the biomedical field: first, in the integration of cancer diagnosis and treatment, MOF-based treatment systems can integrate multiple modes such as chemotherapy, radiotherapy, photodynamic therapy, photothermal therapy, chemodynamic therapy, starvation therapy, and immunotherapy through single or combined strategies to exert a synergistic anti- tumor therapeutic effect; second, by constructing MOF-based carriers, including pH-responsive, GSH-responsive, and photo-responsive carriers, effective loading and precise delivery of drug molecules, including biological macromolecules, can be achieved; third, in the field of in vitro diagnosis, various MOF-based biomarker detection methods have been developed, providing technical means for the precise early diagnosis of diseases. Meanwhile, this paper deeply analyzes the key challenges that MOFs still face in clinical translation, including large-scale preparation, long-term stability, and biological safety assessment, and prospects the future development directions and application prospects.

The formation of protein-protein interaction (PPI) networks is a central event in biochemical reactions within organisms. These interactions not only regulate normal physiological functions but are also closely associated with the onset and progression of diseases. PPIs are intricately regulated by proteins, nucleic acids, and their interactions. The complex molecular networks formed between these molecules serve as the foundation for most biochemical reaction events. Moreover, biological information is transmitted through countless molecular interactions within the cellular environment. A wide range of technologies has been developed to study PPIs, among which proximity-dependent biotinylation is a novel technique for labeling proteomes in living cells. This method utilizes engineered biotin ligases to specifically label nearby proteins or RNA molecules, enabling the capture of transient, weak, or stable interactions and facilitating the systematic construction of molecular interaction maps. Through continuous enzyme optimization and refinement, proximity-dependent biotinylation techniques have evolved into diverse systems with improved operational convenience and labeling efficiency. Each proximity-dependent biotinylation technique offers unique advantages: BioID is non-toxic to cells but suffers from low labeling efficiency, requires 18-24 h for labeling, and yields limited biotinylated products. TurboID achieves efficient labeling within 10 min, but its high activity and strong biotin affinity may lead to cytotoxicity. AirID enables low-toxicity labeling under low biotin concentrations but requires several hours to complete. UltraID offers the highest labeling activity with the smallest molecular mass but is prone to over-labeling. APEX provides convenient operation and can resolve protein topology, yet it has concentration-dependent limitations—forming dimers at high concentrations and lacking sensitivity at low concentrations. RNA-BioID is tailored for studying RNA-protein interactions but is limited by non-specific binding. TransitID can capture dynamic protein translocation at the subcellular level, though its temporal resolution still requires improvement. This review systematically summarizes the development, mechanisms, advantages, and disadvantages of proximity-dependent biotinylation techniques such as BioID, TurboID, AirID, UltraID, RNA-BioID, APEX, and TransitID. It also explores their cutting-edge applications in functional regulation and disease research. Proximity-dependent biotinylation techniques are widely used in disease-related studies. In tumor research, they are primarily applied to investigate the transcriptional regulation and chromosomal structural changes of proto-oncogenes and tumor suppressor genes. In the field of neuroscience, they are used to study mechanisms underlying nervous system function and neurological diseases. In viral infection mechanisms, they help elucidate virus-host interaction networks. In immune regulation, they contribute to the study of immune signaling pathways. In stem cell research, they aid in understanding cell differentiation processes. Furthermore, proximity-dependent biotinylation techniques hold promise for integration with spatial biology technologies, enabling more comprehensive and detailed protein studies. These techniques are expected to provide more accurate and efficient tools for life science research and to advance the medical and health fields to a higher level. By comprehensively analyzing the strengths, limitations, and innovative potential of each method, this review also highlights their advantageous applications in molecular interaction studies, aiming to provide methodological guidance and theoretical support for molecular mechanism research in the life sciences.

Microorganisms, as one of the Earth’s most abundant genetic resources, demonstrate tremendous application potential in fields such as medicine, energy, and environmental protection. However, natural microorganisms often suffer from poor stability and low catalytic efficiency. The emergence of microorganism-nanomaterial hybrid systems offers novel strategies to overcome these limitations. These systems integrate nanomaterials with microorganisms or their components (e.g., cell membranes, metabolites, or biomacromolecules) through methods such as biomineralization, electrostatic assembly, surface modification, and genetic engineering. This enables programmable design from the nanoscale to the macroscale, demonstrating broad application prospects and attracting extensive research interest. First, microbial-nanomaterial hybrid systems are classified based on the types of nanomaterials (organic, inorganic, organic-inorganic) and microorganisms (bacteria, fungi, viruses, algae, probiotics). Both types of systems leverage the unique catalytic selectivity of microorganisms and the diverse physicochemical properties of nanomaterials to achieve multidimensional synergy. Their synergistic mechanisms involve both the biochemical processes of microorganisms and the surface/interface reactions of nanomaterials, representing a multidisciplinary achievement spanning microbial interface engineering, biomimetic catalysis, controllable nanomaterial fabrication, and interfacial transport and reaction processes. Next, the application progress in biomedical fields (such as anti-infection, intestinal diseases, and cancer therapy) and energy conversion (e.g., light-driven hybrid systems for proton reduction to hydrogen, CO₂ reduction and conversion, and nitrogen fixation) is elaborated in detail, highlighting their significant advantages in functional integration and synergistic performance. Microorganism-nanomaterial hybrid systems combine the specific recognition and precise metabolic capabilities of microorganisms with the catalytic, drug-delivery, and optoelectronic functions of nanomaterials, enabling the construction of various multifunctional synergistic platforms for catalysis, diagnosis, and therapy. These advances have greatly promoted development in nanomedicine, energy, and environmental applications. In medical contexts, such systems utilize the natural chemotaxis of microorganisms for precise targeting, achieve controlled drug release through environmentally responsive delivery and metabolic regulation, and enhance therapeutic efficacy via combined chemical-biological treatments and immune modulation. Improved biosafety can be achieved through attenuated microbial designs and nanomaterial coatings, offering diverse strategies for the precise treatment of various diseases. In the energy sector, the excellent light-harvesting properties of semiconductor materials and the precise catalytic capabilities of biological systems have been integrated to successfully construct light-driven biocatalytic systems, significantly improving light utilization efficiency. Finally, this review discusses the key challenges facing the practical application of these systems. Nanomaterials may exert toxic effects on microorganisms, impairing their activity and raising environmental safety concerns. The potential release of engineered nanomaterials into ecosystems necessitates careful risk assessment and long-term monitoring. In real-world environments, microbial functions are easily compromised, nanostructures are prone to damage, and reactive oxygen species (ROS) tend to accumulate, resulting in insufficient system stability. Stringent culture conditions, costly raw materials, and significant batch-to-batch variability hinder large-scale production and commercialization. The synergistic mechanisms between microorganisms and nanomaterials are not yet fully understood, particularly regarding molecular-level interactions and long-term compatibility. In medical applications, off-target risks persist due to unpredictable microbial colonization and immune responses, while environmental applications lack sufficient selective recognition capabilities, indicating a need for improved targeting and specificity. Furthermore, interdisciplinary barriers between biology, materials science, and engineering complicate collaborative innovation, and the absence of well-established standards for evaluation, regulation, and scalability also constrains further development. Future efforts should focus on enhancing biocompatibility, optimizing fabrication processes, and establishing comprehensive safety and performance standards to accelerate the transition of these promising systems from laboratory research to real-world applications.

This review synthesizes recent advances in prolamin-based multicomponent nanocarriers, with a focus on their physicochemical properties, modification strategies, and potential applications in functional foods, biomedicine, and sustainable agriculture. The abundance of hydrophobic amino acid residues in prolamins facilitates spontaneous self-assembly into nanoparticles, making them promising carriers for poorly water-soluble bioactive compounds such as curcumin and resveratrol. However, native prolamin nanoparticles suffer from limitations including poor colloidal stability, tendency to aggregate under processing or physiological conditions (e.g., pH, ionic strength, enzymatic degradation), and limited functional diversity. To address these drawbacks, extensive research has been devoted to modification strategies aimed at enhancing stability, structural integrity, and cargo protection. Polysaccharide modification enables the formation of stable core-shell structures through electrostatic interactions, hydrogen bonding, and steric hindrance. Coatings with pectin, chitosan, or alginate improve stability across a broad range of pH values and ionic strengths, enhance resistance to gastric digestion, and enable sustained release in the intestine, thereby improving bioavailability. Polyphenol modification introduces hydrogen bonding, hydrophobic interactions, and occasionally covalent cross-linking, which modify nanoparticle structure and surface properties. These composites exhibit improved hydrophilicity, colloidal stability, and resistance to oxidative or UV-induced degradation, along with intrinsic antioxidant activity. Lipid modification leverages hydrophobic interactions with oils or fatty acids to form composite nanoparticles or Pickering emulsions. This approach increases the loading capacity for hydrophobic compounds, creates a protective barrier, and enhances oral bioavailability by promoting emulsification and intestinal absorption. Additional strategies include the incorporation of auxiliary proteins (e.g., casein, whey protein) to improve stability and emulsifying capacity, as well as the use of inorganic nanomaterials (e.g., SiO₂, AuNPs) to impart mechanical reinforcement, antibacterial properties, and stimuli-responsive functions. Genetic engineering further allows molecular-level tailoring of amino acid sequences to fine-tune hydrophobicity, amphiphilicity, and self-assembly behavior. These engineered nanocarriers exhibit advanced functionalities. They enable sustained and stimuli-responsive release triggered by pH, redox potential, enzymes, temperature, or light, facilitating on-demand delivery that maximizes efficacy while minimizing off-target effects. Targeting can be achieved passively through the enhanced permeability and retention (EPR) effect, or actively via conjugation with ligands, antibodies, or peptides that recognize specific receptors. The applications of these systems are broad. In functional foods and nutraceuticals, prolamin-based carriers improve the stability, bioavailability, and controlled release of sensitive bioactive ingredients, supporting personalized nutrition. In biomedicine, they enhance oral drug delivery, enable targeted cancer therapy with reduced systemic toxicity, and serve as scaffolds for tissue engineering. In agriculture, they facilitate the controlled release of pesticides, fertilizers, and growth regulators, helping to reduce environmental contamination and promote sustainable practices; they are also being explored for smart food packaging applications. Despite significant progress, challenges remain in clinical and industrial translation. There is an urgent need for standardized characterization methods, comprehensive in vivo safety and efficacy evaluations, and scalable, regulation-compliant manufacturing processes. Future research should adopt rational design principles to develop multi-stimuli-responsive and sustainable systems. The integration of artificial intelligence and data-driven approaches may further accelerate the development of personalized theranostic platforms and co-delivery systems. Continued innovation is expected to solidify the role of prolamin-based nanocarriers in advancing global health and sustainable development.

The comorbidity of sarcopenia and cognitive impairment constitutes a degenerative syndrome that progresses significantly with age. It has emerged as a critical global health challenge, contributing to functional disability, reduced quality of life, and increased pressure on public healthcare systems. This comorbidity is characterized by a synergistic decline in both physical and cognitive capabilities, manifesting as reduced skeletal muscle mass, diminished muscle strength, impaired physical function, and progressive deterioration in cognitive domains such as memory, executive function, and information processing speed. This dual degeneration not only creates a vicious cycle where each condition exacerbates the other but also substantially increases the risk of falls, fractures, hospitalization, and mortality among older adults. Against the backdrop of rapid global population aging, the prevalence of this comorbidity is anticipated to rise further without effective interventions. Consequently, investigating its underlying mechanisms and developing preventive and therapeutic strategies hold substantial clinical and public health significance. Current evidence indicates that the pathogenesis involves multi-system and multi-level pathophysiological processes, with chronic inflammation, mitochondrial dysfunction, and gut microbiota dysbiosis, identified as three core interacting mechanisms. Age-related chronic low-grade inflammation, termed inflammaging, arises from the senescence-associated secretory phenotype (SASP) and persistent immune cell activation. This inflammatory state inhibits the intramuscular IGF-1/Akt/mTOR anabolic pathway through proinflammatory cytokines (e.g., IL-6, TNF-α), while simultaneously activating protein degradation systems including the ubiquitin-proteasome system (UPS) and autophagy-lysosomal pathway (ALP), ultimately leading to accelerated protein breakdown and muscle atrophy. These circulating inflammatory factors can also compromise blood-brain barrier integrity, activate microglia, trigger neuroinflammation, and consequently damage synaptic structures and neuronal function, thereby accelerating cognitive decline in this comorbidity. Mitochondrial dysfunction presents as impaired oxidative phosphorylation efficiency, excessive reactive oxygen species (ROS) production, and dysregulated mitochondrial quality control. This not only results in inadequate cellular energy supply but also enables mitochondrial-derived factors (e.g., extracellular mtDNA) to activate innate immune pathways such as cGAS-STING, propagating stress signals and amplifying tissue damage in both muscle and brain. Additionally, gut microbiota dysbiosis impairs intestinal barrier function, increases lipopolysaccharide (LPS) translocation into circulation, and reduces short-chain fatty acid (SCFA) production. These changes induce systemic inflammation and metabolic disturbances that further impact muscle metabolism and promote pathological protein accumulation in the brain, thereby establishing a gut-brain-muscle axis that exacerbates the progression of this comorbidity. Exerkines represent a class of biologically active signaling molecules—including cytokines, peptides, metabolites, and exosomes—secreted by various tissues in response to exercise. These exerkines mediate systemic adaptations and protective effects through endocrine and paracrine actions on target organs. Key exerkines such as IL-6, irisin, brain-derived neurotrophic factor (BDNF), insulin-like growth factor-1 (IGF-1), fibroblast growth factor-21 (FGF-21), lactate, and cathepsin B (CTSB) play central roles in coordinately ameliorating the comorbidity of sarcopenia and cognitive impairment. The beneficial effects of these exerkines are mediated through multiple mechanisms including inflammation modulation, energy metabolism remodeling, neuroprotection, and enhanced neuroplasticity. As a non-pharmacological intervention, exercise effectively stimulates the production and release of exerkines, thereby targeting the comorbidity through multiple pathways. Aerobic exercise elevates lactate levels and activates the Sirt1/PGC-1α pathway, improving cerebral metabolism and cognitive function. Resistance training significantly upregulates IGF-1, irisin, and CTSB expression, enhancing muscle anabolism and hippocampal function. Other modalities like high-intensity interval training (HIIT) and traditional practices also help modulate inflammatory status and optimize the neurotrophic environment through the action of various exerkines. Different exercise types work synergistically by engaging distinct signaling pathways and exerkine combinations, collectively alleviating chronic inflammation, correcting mitochondrial dysfunction, and optimizing gut microecology to achieve concurrent musculoskeletal and cognitive protection against this comorbidity. Synthesizing current evidence, this review emphasizes the necessity of transcending a single-organ perspective by recognizing muscle and brain as an integrated functional unit, with exerkines playing a pivotal role in the muscle-brain axis. The field nevertheless faces several challenges: the secretion dynamics of exerkines during aging remain unclear, mechanisms underlying individual differences in exercise response require elucidation, and the compensatory and imbalance characteristics of exercise-induced exerkine networks across disease stages need further characterization. Future research should employ large-sample cohorts and randomized controlled trials integrated with multi-omics technologies to establish personalized exercise interventions based on exerkine profiling for managing this comorbidity. Parallel efforts should focus on developing quantifiable efficacy assessment systems to provide robust theoretical foundation and practical guidance for precise management of the comorbidity of sarcopenia and cognitive impairment and the promotion of healthy aging.

Olfactory receptors (ORs) form the largest superfamily of G protein-coupled receptors (GPCRs). Traditionally recognized for their role in the nasal olfactory epithelium, where they mediate the sense of smell, accumulating evidence has firmly established their ectopic expression in non-olfactory tissues, including the intestine, lungs, and kidneys. The intestine, as the primary site for nutrient digestion and absorption, harbors a highly complex chemical environment. To adapt to this environment, the gut employs a sophisticated network of "chemosensors" to monitor luminal contents and maintain homeostasis. Among these sensors, intestinal ORs have emerged as crucial functional components, serving as a molecular bridge that connects environmental chemical signals—such as food-derived odorants—to specific physiological responses. This discovery has significantly deepened our understanding of how dietary flavors and compounds influence intestinal physiology at the molecular level. This review systematically summarizes the expression profiles, ligand classification, and biological functions of ORs within the gastrointestinal tract. Studies indicate that intestinal ORs exhibit distinct spatial distribution patterns across different gut segments and display cell-type specificity, particularly within enterocytes and enteroendocrine cells. These receptors function as versatile sensors capable of recognizing a wide variety of ligands, including exogenous dietary components, gut microbiota metabolites such as short-chain fatty acids, and endogenous small molecules like azelaic acid. Upon activation by specific ligands, intestinal ORs trigger intracellular signaling cascades, primarily involving the AC-cAMP-PKA pathway or calcium influx channels. A major focus of this review is to elucidate the molecular mechanisms by which these receptors regulate the secretion of gut hormones. Activation of specific ORs in enteroendocrine cells has been shown to stimulate the release of hormones such as glucagon-like peptide-1 (GLP-1), peptide YY (PYY), and serotonin (5-HT), thereby modulating systemic energy metabolism, glucose homeostasis, and gastrointestinal motility. Furthermore, the review addresses the critical roles of ORs in immune regulation and pathology. Evidence suggests that specific ORs contribute to the maintenance of intestinal immune homeostasis and may offer protection against inflammation. Beyond their involvement in inflammatory responses, ORs such as Olfr78 have been shown to regulate the differentiation and function of intestinal endocrine cells. Similarly, Olfr544 has been demonstrated to alleviate intestinal inflammation by remodeling the gut microbiome and metabolome. These findings collectively suggest that specific ORs hold promise as therapeutic targets for mitigating intestinal inflammation and maintaining gut homeostasis. Additionally, the review explores the emerging role of ORs in cancer. Although OR expression is often downregulated in tumor tissues compared to normal mucosa, activation of specific ORs by certain ligands can inhibit tumor cell proliferation and migration and induce apoptosis via pathways such as MEK/ERK and p38 MAPK. Conversely, other receptors, such as OR7C1, may serve as biomarkers for cancer-initiating cells. In conclusion, intestinal ORs represent a vital component of the gut"s sensory network. The review also discusses the translational potential of these findings. By elucidating the precise pairing relationships between dietary components and specific ORs, novel therapeutic strategies could be developed. Intestinal ORs may thus emerge as promising targets for nutritional and pharmacological interventions in metabolic diseases, inflammatory bowel diseases, and malignancies.

Objective Pancreatic ductal adenocarcinoma (PDAC) exhibits a limited response to current treatments due to its dense fibrotic stroma and highly immunosuppressive tumor microenvironment. In recent years, advancements in cellular immunotherapy, particularly chimeric antigen receptor macrophage (CAR-M) therapy, have offered new hope for pancreatic cancer treatment. Although CAR-M therapy demonstrates dual potential in directly killing tumor cells and remodeling the immune microenvironment, it still faces challenges such as complex in vitro preparation processes and low in vivo targeting and delivery efficiency. Therefore, developing strategies for efficient and targeted in vivo delivery of CAR genes has become crucial for overcoming current therapeutic limitations. This study aims to develop an orally administrable nano-gene delivery system for the targeted delivery of CAR genes to pancreatic tumor sites.Methods Core nano-gene particles (PNP/pCAR) were constructed by loading plasmid DNA encoding CAR (pCAR) with cationic polypeptides (PNP). Subsequently, PNP/pCAR was surface-modified with β-glucan to prepare the targeted nanoparticles (βGlus-PNP/pCAR). The loading efficiency of PNP for pCAR was quantitatively assessed by gel retardation assay. The particle size, Zeta potential, morphology, and storage stability of PNP/pCAR were characterized using a Malvern particle size analyzer and transmission electron microscopy. At the cellular level, RAW 264.7 macrophages were selected. The cytotoxicity of PNP/pCAR was evaluated using the CCK-8 assay. The cellular uptake efficiency and lysosomal escape ability of the nanoparticles were assessed via flow cytometry and confocal microscopy. Transfection efficiency was quantitatively evaluated by detecting the expression of the reporter gene GFP using flow cytometry. At the in vivo level, an orthotopic pancreatic cancer mouse model was established. Cy7-labeled βGlus-PNP/pCAR nanoparticles were administered orally, and the fluorescence distribution in mice was dynamically monitored at 1, 2, 4, 8, and 16 hours post-administration using a small animal in vivo imaging system. Forty-eight hours after oral gavage, the mice were euthanized, and pancreatic tumor tissues were collected for further analysis of intratumoral fluorescence signals using the imaging system. Additionally, βGlus-PNP/pCAR-GFP nanoparticles loaded with the reporter gene (GFP) were administered orally. Forty-eight hours post-administration, pancreatic tumor tissues were harvested to prepare frozen sections, and GFP expression was observed and analyzed under a fluorescence microscope.Results The PNP carrier exhibited a high loading capacity for pCAR. The successfully prepared PNP/pCAR nanoparticles were regular spheres with a hydrodynamic diameter of approximately (120 ± 10) nm and a Zeta potential of about +(6 ± 1) mV. They maintained good structural stability after incubation in PBS buffer for 7 days. Cell experiments demonstrated that PNP/pCAR exhibited no significant cytotoxicity in RAW 264.7 cells while being efficiently internalized and effectively escaping lysosomal degradation. The transfection positive rate of PNP/pCAR-GFP in RAW 264.7 cells reached (25 ± 3)%, surpassing that of Lipofectamine 2000-loaded pCAR-GFP (Lipo/pCAR-GFP), which was (20 ± 1)%. In vivo experiments revealed that, compared to unmodified PNP/pCAR, βGlus-PNP/pCAR exhibited stronger in situ pancreatic tumor targeting ability after oral administration. Furthermore, oral administration of βGlus-PNP/pCAR-GFP resulted in significant GFP protein expression detectable within pancreatic tumor tissues.Conclusion This study successfully constructed and validated an orally administrable, pancreatic cancer-targeting polypeptide-based nano-gene delivery system. It provides an important technological foundation in delivery systems and experimental basis for the subsequent development of in situ CAR-M-based therapeutic strategies for pancreatic cancer.

Objective Cleft palate (CP) is a common congenital deformity often associated with velopharyngeal insufficiency (VPI), which disrupts the physiological coupling between respiration and speech. Conventional clinical assessments, such as nasometry and spirometry, provide limited static data and fail to visualize the dynamic spatiotemporal distribution of lung ventilation during phonation. This study introduces spatiotemporal electrical impedance tomography (ST-EIT) to evaluate speech-respiratory functional features in CP patients compared to normal controls (NC). The aim is to characterize multi-domain respiratory patterns and to validate an interpretable machine learning framework for providing objective, quantitative evidence for clinical assessment.Methods Seventy-five participants were enrolled in this study, comprising 37 patients with surgically repaired CP and 38 healthy volunteers matched for age, gender, and body mass index (BMI). All subjects performed standardized sustained phonation tasks while undergoing synchronous monitoring with a 16-electrode EIT system and a pneumotachograph. A comprehensive feature engineering pipeline was developed to extract physiological parameters across three complementary domains: (1) Temporal domain: including inspiratory/expiratory phase duration (tPhase), time constants (Tau), and inspiratory-to-expiratory time ratios (TI/TE); (2) Airflow domain: comprising mean flow, peak flow, and instantaneous flow at 25%, 50%, and 75% of tidal volume; and (3) Spatial domain: quantifying global and regional tidal impedance variation (TIV), global inhomogeneity (GI), and center of ventilation (CoV). Extreme Gradient Boosting (XGBoost) classifiers were trained using five distinct data sources (Spirometry, Nasometry, Inspiratory-EIT, Expiratory-EIT, and fused ST-EIT). Model performance was rigorously evaluated via stratified 5-fold cross-validation, and Shapley Additive Explanations (SHAP) were employed to quantify global and local feature contributions.Results The CP group exhibited a distinct respiratory phenotype compared to controls. In the temporal domain, CP patients showed significantly shorter inspiratory (1.60 vs. 1.85 s, P<0.001) and expiratory phase durations (2.45 vs. 3.95 s, P<0.001), indicating a rapid, shallow breathing rhythm. In the airflow domain, while inspiratory flows were comparable, the CP group demonstrated significantly elevated mean and peak flows during the expiratory phase (P<0.001), reflecting compensatory respiratory effort. Spatially, CP patients presented significant ventilation redistribution, characterized by higher regional TIV in the right-anterior (ROI1) and left-posterior (ROI4) quadrants, but lower TIV in the left-anterior (ROI2) quadrant. In terms of diagnostic accuracy, the multi-modal ST-EIT model achieved the highest performance (AUC: 0.915 ± 0.012, Accuracy: 0.843 ± 0.019, F1-score: 0.872 ± 0.017), substantially outperforming models based on spirometry (AUC: 0.721) or nasometry (AUC: 0.625) alone. Interpretability analysis revealed that spatial domain features were the most critical, contributing 53.4% to the model"s decision-making, followed by temporal (25.0%) and airflow (21.6%) features.Conclusion ST-EIT successfully captures the temporal, airflow, and spatial deviations in CP speech respiration that are undetectable by conventional methods—specifically, rapid phase transitions, hyperdynamic expiratory airflow, and regional ventilation heterogeneity. This study validates ST-EIT as a robust, non-invasive, and radiation-free tool for characterizing speech-respiratory dysfunction, offering high clinical value for bedside screening, rehabilitation planning, and longitudinal monitoring of patients with cleft palate.

Osteoarthritis (OA), a highly prevalent degenerative joint disease worldwide, is defined by articular cartilage degradation, abnormal bone remodeling, and persistent chronic inflammation. It severely compromises patients’ quality of life, and currently, there is no radical cure. Abnormal mechanical stress is widely regarded as a core driver of OA pathogenesis, and the exploration of mechanical signal perception and transduction mechanisms has become crucial for deciphering OA’s pathophysiological processes. Piezo1, a key mechanosensitive cation channel belonging to the Piezo protein family, has recently gained significant attention due to its pivotal role in mediating cellular responses to mechanical stimuli in joint tissues. This review systematically examines Piezo1’s expression patterns, regulatory mechanisms, and pathological functions in OA, with a particular focus on its dual roles in modulating chondrocyte homeostasis and bone metabolism disorders, while also delving into the underlying molecular signaling pathways and potential therapeutic implications. Piezo1, consisting of approximately 2 500 amino acids and forming a unique trimeric propeller-like structure, is widely expressed in chondrocytes, osteocytes, mesenchymal stem cells, and synovial cells. It exhibits permeability to cations such as Ca²⁺, K⁺, and Na⁺, and directly responds to membrane tension changes induced by mechanical stimuli like fluid shear stress and mechanical overload. In OA patients and animal models, Piezo1 expression is significantly upregulated, especially in cartilage regions subjected to abnormal mechanical stress (e.g., human temporomandibular joint cartilage). This overexpression is closely associated with aggravated cartilage degeneration, increased chondrocyte apoptosis, accelerated cellular senescence, and intensified inflammatory responses. Mechanical overload and pro-inflammatory cytokines (e.g., IL-1β) are key inducers of Piezo1 upregulation: IL-1β activates the PI3K/AKT/mTOR signaling pathway to enhance Piezo1 expression, forming a pathogenic positive feedback loop that inhibits chondrocyte autophagy, promotes apoptosis, and further accelerates joint degeneration. Mechanistically, Piezo1 mediates OA progression through multiple interconnected pathways. When activated by mechanical stress, Piezo1 triggers excessive Ca²⁺ influx, leading to endoplasmic reticulum stress (ERS) and mitochondrial dysfunction, which directly induce chondrocyte apoptosis. This process involves the activation of downstream signaling cascades such as cGAS-STING and YAP-MMP13/ADAMTS5. YAP, a transcriptional regulator, upregulates the expression of matrix metalloproteinase 13 (MMP13) and aggrecanase (ADAMTS5), thereby accelerating cartilage matrix degradation. Additionally, Piezo1-driven Ca²⁺ overload promotes the accumulation of reactive oxygen species (ROS) and upregulates senescence markers (p16 and p21), accelerating chondrocyte senescence via the p38MAPK and NF-κB pathways. Senescent chondrocytes secrete senescence-associated secretory phenotype (SASP) factors (e.g., IL-6, IL-1β), further amplifying joint inflammation. In terms of bone metabolism, Piezo1 maintains joint homeostasis by promoting the differentiation of fibrocartilage stem cells into chondrocytes and balancing bone formation and resorption through regulating the FoxC1/YAP axis and RANKL/OPG ratio. Therapeutically, targeting Piezo1 shows promising potential. Preclinical studies have demonstrated that Piezo1 inhibitors (e.g., GsMTx4) can reduce joint damage and alleviate pain in OA mice. Simultaneously, siRNA-mediated co-silencing of Piezo1 and TRPV4 (another mechanosensitive channel) decreases intracellular Ca²⁺ concentration, inhibits chondrocyte apoptosis, and promotes cartilage repair. Conditional knockout of Piezo1 using Gdf5-Cre transgenic mice alleviates cartilage degeneration in post-traumatic OA models by downregulating MMP13 and ADAMTS5 expression. Despite existing challenges, such as off-target effects of inhibitors, inefficient local drug delivery, and interindividual genetic variability, strategies like developing selective Piezo1 antagonists, optimizing targeted nanocarriers, and combining Piezo1-targeted therapy with physical therapy provide viable avenues for clinical translation. The authors propose that Piezo1 serves as a critical therapeutic target for OA, and future research should focus on deciphering its context-dependent regulatory networks, developing tissue-specific intervention strategies, and validating their efficacy and safety in clinical trials to address the unmet medical needs of OA patients.

Organoid-on-a-chip technology represents a promising interdisciplinary advancement that merges two cutting-edge biomedical platforms: stem cell-derived organoids and microfluidics-based organ-on-a-chip systems. Organoids are self-organizing three-dimensional (3D) cell cultures that mimic the key structural and functional features of in vivo organs. However, traditional organoid culture systems are often static, lacking dynamic environmental cues and suffering from limitations such as batch-to-batch variability, low stability, and low throughput. Organ-on-a-chip platforms, by contrast, utilize microfluidic technologies to simulate the dynamic physiological microenvironment of human tissues and organs, enabling more controlled cell growth and differentiation. By integrating the advantages of organoids and organ-on-a-chip technologies, organoid-on-a-chip systems transcend the limitations of conventional 3D culture models, offering a more physiologically relevant and controllable in vitro platform. In organoid-on-a-chip systems, stem cells or pre-formed organoids are cultured in micro-engineered environments that mimic in vivo conditions, enabling precise control over fluid flow, mechanical forces, and biochemical cues. Specifically, these platforms employ advanced strategies including bio-inspired 3D scaffolds for structural support, precise spatial cell patterning via 3D bioprinting, and integrated biosensors for real-time monitoring of metabolic activities. These synergistic elements recreate complex extracellular matrix signals and ensure high structural fidelity. Based on structural complexity, organoid-on-a-chip systems are classified into single-organoid and multi-organoid types, forming a trajectory from unit biomimicry to systemic simulation. Single-organoid chips focus on highly biomimetic units by integrating vascular, immune, or neural functions. Multi-organoid chips simulate inter-organ crosstalk and systemic homeostasis, advancing complex disease modeling and PK/PD evaluation. This emerging technology has demonstrated broad application potential in multiple fields of biomedicine. Organoid-on-a-chip systems can recapitulate organ development in vitro, facilitating research in developmental biology. They mimic organ-specific physiological activities and mechanisms, showing promising applications in regenerative medicine for tissue repair or replacement. In disease modeling, they support the reconstruction of models for neurodegenerative, inflammatory, infectious, metabolic diseases, and cancers. These platforms also enable in vitro drug testing and pharmacokinetic studies (ADME). Patient-derived chips preserve genetic and pathological features, offering potential for precision medicine. Additionally, they reduce species differences in toxicology, providing human-relevant data for environmental, food, cosmetic, and drug safety assessments. Despite progress, organoid-on-a-chip systems face challenges in dynamic simulation, ECM variability, and limited real-time 3D imaging, requiring improved materials and the integration of developmental signals. Current bottlenecks also include the high technical threshold for automation and the lack of standardized validation frameworks for regulatory adoption. Meanwhile, the concept of a "human-on-a-chip" has been proposed to mimic whole-body physiology by integrating multiple organoid modules. This approach enables systemic modeling of drug responses and toxicity, with the potential to reduce animal testing and revolutionize drug development. Future advancements in bio-responsive hydrogels and flexible biosensors will further empower these platforms to bridge the gap between bench-side research and personalized clinical interventions. In conclusion, organoid-on-a-chip technology offers a transformative in vitro model that closely recapitulates the complexity of human tissues and organ systems. It provides an unprecedented platform for advancing biomedical research, clinical translation, and pharmaceutical innovation. Continued development in biomaterials, microengineering, and analytical technologies will be essential to unlocking the full potential of this powerful tool.

RNA synthetic biology, as a frontier interdisciplinary field, is driving the leap from fundamental research to precision medicine in life sciences through the engineered design of RNA components and the construction of genetic circuits. This paper aims to systematically outline the design principles, key technological breakthroughs, and biomedical applications of synthetic RNA genetic circuits. Building upon this foundation, it provides an in-depth analysis of current research bottlenecks and proposes future development directions. Commencing with a foundational role in the central dogma of RNA, this paper establishes a systematic classification framework for synthetic biology RNA components. At the cis-acting element level, it elaborates on how components such as riboswitches, RNA thermometers, and Toehold switches achieve precise gene expression regulation by responding to specific ligands, temperatures, or trigger RNAs through conformational changes. Concerning trans-acting elements, it delves into the molecular mechanisms of miRNA-mediated gene silencing, the high stability and "sponge-like adsorption" function conferred by the closed-loop structure of circRNA, the targeting role of siRNA within the RNAi pathway, and the targeting specificity of sgRNA within the CRISPR system. The research emphasizes that rational design, sequence optimization, and chemical modifications can significantly enhance the performance and orthogonality of these natural elements. Secondly, the paper focuses on the design and optimization strategies for synthetic RNA regulatory modules. Taking miRNA-responsive circRNA switches as an example, it elucidates the principles of customized miRNA responsiveness. The engineering applications of circRNA are explored, introducing strategies for constructing functional RNA nanostructures via siRNA self-assembly. Building upon this, the paper emphasizes synthetic genetic circuits: from logical operations to resource allocation, enabling advanced cellular logic and functional regulation. For instance, by combining transcriptional cascade switches or utilizing the CRISPR-Cas13a system, an AND logic gate responsive to multiple miRNAs (such as miRNA-155 and miRNA-21) was constructed, significantly enhancing the specificity of disease diagnosis. Addressing the challenges of resource competition and expression noise faced by synthetic circuits within cells, this paper introduces computational models such as MIRELLA, with particular emphasis on the design of endogenous miRNA-based iFFLs. These advanced circuits, illustrated in this paper, have been successfully applied to real-time monitoring of cellular differentiation states and regulation of stem cell-directed differentiation. For cellular state detection and dynamic regulation, miRNA switches can be integrated with fluorescent systems to track differentiation statuses in real time via fluorescent signal changes. Synthetic genetic circuits, meanwhile, utilize endogenous miRNA logic integration alongside miSFITs technology to achieve state-specific protein regulation in human pluripotent stem cells, laying the groundwork for customized cellular control. This approach ingeniously harnesses intrinsic cellular regulatory mechanisms to buffer gene expression burdens, thereby enhancing circuit robustness. These advanced circuits, illustrated schematically herein, have been successfully applied to real-time monitoring of cellular differentiation states and regulation of stem cell-directed differentiation. At the therapeutic translation level, the paper systematically reviews application strategies for RNA technologies across multiple fields, including cancer, metabolic diseases, neurodegenerative diseases, cardiovascular diseases, regenerative medicine engineering, immunotherapy, and vaccine applications. For instance, in cancer treatment, specific killing of tumor cells is achieved by embedding targets for miRNAs specific to healthy cells within the genomes of oncolytic viruses (such as Zika virus). Within metabolic and degenerative diseases, LNP-delivered mRNA therapeutics and antisense oligonucleotide (ASO) technologies have demonstrated significant clinical progress. Finally, this paper highlights ongoing challenges in the field, including limited programmability of RNA elements, low in vivo delivery efficiency, and inadequate off-target risk assessment systems. It advocates for future integration of epigenomics and computational modelling to optimize element functionality, establishing an integrated "element-circuit-delivery" platform. Furthermore, leveraging single-cell sequencing and organoid technologies to develop a multidimensional safety assessment system is proposed to advance the deep integration and translation of RNA synthetic biology in personalized medicine. Consequently, RNA engineering has transcended single-dimensional regulation, evolving towards multi-layered, dynamic, and intelligent synthetic biological systems. Its deep integration with clinical needs will reshape disease diagnosis and treatment paradigms.

Objective To clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology.Methods The ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture.Results Behaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01).Conclusion METTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of "opening Xuanfu and dredging myelin".

Depressive disorder is a prevalent mental illness characterized by pronounced and enduring symptoms of depression and cognitive impairment. The escalating pressures of modern society have led to a corresponding rise in the number of depressive disorder patients, particularly those exposed to adverse social, economic, political, and environmental factors which exacerbate the risk of this disorder. The pathogenesis of depressive disorder is multifaceted, encompassing oxidative stress, neuroplasticity alterations, neuroinflammation, neurotransmitter system imbalances, and intestinal microecological disruptions, among others. Clinically, conventional antidepressants are primarily predicated on the monoamine neurotransmitter hypothesis. This theory posits that depressive disorder can be ameliorated by regulating the levels of neurotransmitters within the body through a singular mechanism. However, the complex and multifaceted pathogenesis of depressive disorder results in limited selectivity for these drugs. Mitogen-activated protein kinase (MAPK) is a conserved serine/threonine kinase that plays a crucial role in various cellular physiological and pathological processes, including cell growth, differentiation, stress adaptation, and inflammatory response. It is instrumental in maintaining cellular homeostasis and regulating cellular responses. Numerous studies indicate MAPK is involved in the pathogenesis and progression of depressive disorder through various pathogenesis. However, what deserves attention is that the interaction between the pathogenesis and dynamics of regulatory process remains unclear. Modulating MAPK has been shown to influence the onset and progression of depressive disorder, though the precise mechanism remains elusive. Within the MAPK family, aberrant activity of extracellular signal-regulated kinase (ERK) can damage hippocampal neurons and overactivate microglia, precipitating depressive disorder. Excessive activation of c-Jun N-terminal kinase (JNK) results in heightened neuronal apoptosis in the hippocampus and prefrontal cortex, and suppresses the expression of neurotrophic factors. p38, a key regulator in inflammatory reactions, can induce neuroinflammation when overactive, leading to depressive disorder. ERK, JNK, and p38 sub-pathways do not function in isolation but rather interact synergistically and/or antagonistically through shared activators and common target molecules. Consequently, these sub-pathways form a complementary and coordinated regulatory network. In addition, MAPK family members can jointly influence the process of depressive disorder by sharing upstream factors and regulating common downstream targets, and there is a lack of identification of their markers and screening for subgroups. The collective abnormal activities of these MAPK family members illuminate the underlying mechanisms of depressive disorder, suggesting that MAPK could serve as a potential therapeutic target for this disorder. As for the study of ERK, different models of depressive disorder have contradictory effects on its activity. The primary cause of these differences can be attributed to the distinct pathological environments utilized in the creation of depressive disorder models. In the future, it is suggested that we use the inducement of depressive disorder as a modeling standard to accurately simulate the onset of depressive disorder to carry out accurate treatment according to the causes of depressive disorder. Research shows that classic clinical drugs, novel MAPK inhibitors and certain traditional Chinese medicines can prevent and treat depressive disorder by regulating the MAPK signaling pathway. Research on MAPK remains limited, particularly concerning the permeability and cellular specificity across the blood-brain barrier and the identification of objective predictive markers. Although inhibitors face challenges, they also possess significant advantages and developmental potential. This paper systematically summarizes the current status of MAPK in the treatment of depressive disorder, in order to provide insights for researching the pathogenesis of depressive disorder and developing new antidepressant drugs.

Objective This study aims to explore the potential of different orders of magnitude SNP locus combinations for predicting distant kinship relationships. A high-density SNP locus set was constructed, and a comprehensive assessment of its inference capability was conducted.Methods Firstly, we selected three commercial chip panels, CGA (Chinese genotyping array, Illumina), GSA (Global screening array, Illumina), Affy (23MF_V2 high-density SNP array, Affymetrix) and merged them after quality control, forming a high-density SNP locus panel(1 180 k). Secondly, we selected 161 samples and collected their peripheral blood samples by using whole-genome sequencing technology. Within this sample population, the levels of kinship relationships fully covered the range from level 1 to level 9, and the number of kinship pairs at each level was consistently maintained at over 50 pairs. From 161 samples data of whole-genome sequencing, the 1 180 k locus set was extracted, which is referred to as the high - density SNP locus set in the following text. The kinship inference was conducted using the Identity-By-Descent (IBD) segment algorithm with the selected optimal parameters. To comprehensively evaluate the performance of the high-density SNP locus set in kinship inference, we compared it with the three commercial chip panels, the intersection of these three chip loci, and the control sets constructed by randomly reducing the number of the high-density SNP locus set. Based on the changes in the IBD segment lengths, as well as the dynamic trends in prediction accuracy, we conducted a scientific assessment of the kinship inference capability of the high-density SNP locus set.Results After screening, a set of 1 184 334 autosomal SNPs was obtained. During the process of screening the optimal IBD segment threshold, the result revealed that 0 cM, 1 cM, and 2 cM all demonstrated good applicability. However, to avoid the issue of a large amount of redundant information caused by setting a too low segment length threshold, this study ultimately selected 2 cM as the optimal threshold. Compared with the average results of three chip panels, the high-density SNP locus set increased the total IBD fragment length and the average IBD fragment length across levels 1-9; the accuracy of the confidence interval for level 8 was 70.97%, which represented a 3.50% improvement; the average confidence interval accuracy for levels 1-8 was 91.39%, representing a 1.00% increase; and the false negative rates at levels 8 and 9 were reduced by 2.42% and 6.76%, respectively. The system efficacy of the high-density SNP locus set for kinship inference of first to eighth degree relationships reached 98.91%. Through random reduction of the high-density SNP locus set results, it is found that increasing the number of SNPs panel, the detection efficiency of IBD segment length showed a significant upward trend. At the same time, the overall trend in the accuracy of kinship relationship prediction as well as the confidence interval accuracy also indicated that both metrics steadily increased with the addition of more loci.Conclusion The results show that the high-density SNPs panel significantly enhances the efficacy of distant kinship inference, accurately covering kinship degrees, with the average confidence interval accuracy for first to eighth degree relationships stably above 90%. The study finds that increasing the number of SNPs panel can improve the ability to predict distant kinship.

Working memory (WM) serves as the core of advanced cognitive functions, enabling the temporary storage and manipulation of information, which is crucial for reasoning, comprehension, and decision-making. However, its performance is influenced by emotional states; for instance, stress or anxiety may impair accurate recall or prioritize the processing of threatening stimuli. This review integrates research on the neural oscillatory mechanisms by which emotions affect WM, emphasizing shared patterns in the θ, α, β, and γ frequency bands. Emotions activate distinct neural circuits and alter oscillatory characteristics through arousal and valence, overlapping with the neural activities required for WM processes. This article aims to elucidate these mechanisms and propose a dual-pathway theoretical framework for emotional influences on WM. The cognitive efficiency hypothesis posits that emotions and WM compete for shared oscillatory resources. Both emotional processing and high WM load enhance cortical excitability by reducing α power to optimize attention allocation. Negative emotions, under sufficient presentation time, prioritize resource allocation to improve WM precision through enhanced sustained α suppression, albeit at the cost of reducing the number of remembered items. The patterns of θ power increases induced by emotions and task load overlap, potentially leading to θ/β saturation under high load, which limits cognitive regulation, while under low load, they synergistically support information maintenance. Emotional states and the β oscillations relied upon for WM maintenance converge in frequency, prone to synergy or competition. High arousal induced by emotions, as well as anxiety alleviation through music, can reduce prefrontal β oscillation power, bringing the brain closer to the low-β state required for WM maintenance, thereby enhancing memory performance and neural efficiency. Similar γ oscillations and α-γ coupling patterns induced by emotions and WM compete for neural resources, interfering with inter-brain region information integration and weakening the regulation of γ amplitude by α rhythms, thus impairing information exchange efficiency and processing stability. The interference hypothesis suggests that emotions directly disrupt the rhythms required for WM. Negative emotions typically reduce the power and synchrony of α oscillations, a change opposite to the α enhancement mode needed during the WM maintenance phase, thereby weakening interference shielding capabilities. The decrease in θ oscillation power induced by negative emotions contrasts with the θ enhancement required for WM tasks, thereby interfering with information maintenance, manipulation, and multi-item integration, leading to declines in memory capacity and precision. Negative or high-arousal emotions significantly disrupt β oscillation power and synchrony in WM tasks, resulting in impaired inter-brain coordination, reduced information stability, and consequently weakened task performance. Emotions directly alter γ oscillation power and θ-γ coupling patterns, with these changes opposing the enhancement direction required for WM tasks, thereby causing neural activity imbalances and information integration obstacles. The cognitive efficiency hypothesis and the interference hypothesis, as a dual-pathway model for emotional influences on WM, are not opposing explanatory frameworks but rather reflect a dynamic and complementary regulatory mechanism in emotion-cognition interactions. The brain flexibly selects interference or efficiency pathways based on current task load, emotional intensity, and individual states to maintain overall functional stability, with its core mechanism lying in the limited and adaptive allocation of brain neural oscillatory resources.Future research should delve into the impacts of emotional states, cognitive load, emotional arousal, and regulatory strategies on WM and its neural modulatory effects, to optimize personalized cognitive intervention strategies.

Objective Using fecal microbiota transplantation (FMT), we established a stroke rat model to investigate the interplay between gut microbiota dysbiosis and ischemic stroke.Methods A preliminary experiment was conducted to establish an antibiotic-induced pseudo-sterile (ABX) rat model through antibiotic treatment, and a cerebral ischemia model was prepared using the middle cerebral artery occlusion (MCAO) method. Fecal microbiota from stroke patients and healthy individuals were transplanted via FMT, followed by behavioral testing. 16S rRNA sequencing was used to analyze the microbial community, hematoxylin and eosin (HE) staining to observe histopathological status, transmission electron microscopy (TEM) to examine the tight junction structure of the small intestine, and enzyme-linked immunosorbent assay (ELISA) to detect levels of inflammatory factors and intestinal barrier-related markers.Results 16S rRNA sequencing of fecal samples showed that compared with the normal control group and the metronidazole group, the abundance and diversity of fecal microorganisms in the quadruple antibiotic group were significantly reduced, indicating successful establishment of the ABX model. After transplanting fecal microbiota from stroke patients into ABX rats, significant changes in gut microbiota composition were observed. Behavioral tests revealed that the MCAO model group showed significant decreases in both horizontal movement and vertical exploration abilities. ELISA results indicated that IL-17 concentration in the ABX+mFMT group was lower than in the ABX+cFMT group, suggesting that IL-17 may serve as a key inflammatory indicator for evaluating the impact of stroke intervention on gut microbiota. TTC staining suggested that gut microbiota intervention may increase the risk of stroke. HE staining showed that, except for the control group, all groups exhibited ischemic changes and inflammatory infiltration in brain tissues. TEM revealed that microvilli of small intestinal epithelial cells in the ABX+mFMT group were sparser than those in the ABX+cFMT group, indicating that microbial intervention affects intestinal barrier function.Conclusion The ABX model established using broad-spectrum antibiotics showed no significant differences in physiological characteristics compared to normal rats, and the findings were consistent with those from germ-free rat models. Stroke prognosis appears to be influenced by intestinal dysbiosis, accompanied by significantly elevated levels of the pro-inflammatory cytokine IL-17, which may exacerbate neural injury via the gut-brain axis. Behavioral experiments indicated that transplantation of gut microbiota from stroke rats impaired cognitive function. Furthermore, IL-17 demonstrated sensitivity to alterations in the gut microbiota, suggesting its potential as a key therapeutic target for stroke intervention.

Objective Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS); however, its underlying neurological pathogenic mechanisms remain incompletely understood. Endogenous formaldehyde (FA), a metabolic byproduct of methylation-demethylation cycles, has recently been implicated in neurotoxicity, oxidative damage, and cognitive impairment. This study aimed to investigate whether excessive FA contributes to myelin sheath demyelination in mice and to evaluate the protective effects and mechanisms of two FA-elimination strategies: sodium bisulfite (NaHSO₃), a classical FA scavenger, and polyethylene glycol-modified astaxanthin nanoparticles (PEG-ATX@NPs), a brain-targeted nano-antioxidant formulation.Methods A chronic demyelination model was established by feeding female C57BL/6J mice a diet containing 0.2% cuprizone (CPZ) for four weeks, followed by a two-week intervention period. Eighty mice were randomly assigned to four groups: NS (normal saline), CPZ+NS, CPZ+NaHSO₃, and CPZ+PEG-ATX@NPs. Behavioral tests, including open-field, Y-maze, and pole-climbing assays, were conducted to assess locomotor activity, motor coordination, and working memory. FA levels in serum, corpus callosum, and spinal cord were measured using an Na-FA fluorescent probe and quantified via in vivo and ex vivo fluorescence imaging. Neuroinflammatory responses were evaluated by measuring TNF-α, IL-1β, and IL-6 levels using ELISA, while oxidative stress was assessed by reactive oxygen species (ROS) fluorescence intensity. Demyelination was examined via Luxol fast blue staining, and microglial activation was analyzed by Iba1 immunofluorescence. Correlation analyses were performed to explore relationships among FA levels, inflammatory cytokines, ROS intensity, and behavioral parameters.Results Compared with the NS group, mice in the CPZ+NS group exhibited significant weight loss, impaired motor coordination and memory, and markedly reduced myelin regeneration (P<0.05). FA levels and pro-inflammatory cytokines were significantly elevated in serum, corpus callosum, and spinal cord (P<0.05). FA-associated fluorescence in brain and spinal tissues, as well as ROS intensity across all tissues examined, also increased substantially (P<0.05). CPZ treatment induced pronounced microglial activation and severe demyelination in the corpus callosum (P<0.01). Both NaHSO₃ and PEG-ATX@NPs effectively reduced FA accumulation in the brain and spinal cord, attenuated demyelination, suppressed microglial activation, decreased inflammatory cytokine levels, and improved motor and cognitive performance. These results confirm that CPZ induced severe demyelination accompanied by oxidative stress, neuroinflammation, and abnormal FA accumulation. Following intervention with either NaHSO₃ or PEG-ATX@NPs, endogenous FA levels in the CNS were substantially reduced. Both treatments alleviated demyelination and significantly decreased the number of activated microglia. Levels of TNF-α, IL-1β, and IL-6 in serum, corpus callosum, and spinal cord were downregulated. Behavioral performance improved significantly, as evidenced by enhanced locomotor activity, better coordination, and improved memory function. These findings indicate that both FA-scavenging agents mitigate CPZ-induced biochemical and behavioral abnormalities.Conclusion This study demonstrates that excessive endogenous FA is closely associated with cognitive impairment, inflammatory dysregulation, and demyelination in a CPZ-induced chronic demyelination mouse model. Clearing abnormally elevated FA effectively reduces neuroinflammation, suppresses microglial overactivation, decreases oxidative stress, and alleviates demyelination, ultimately improving motor and cognitive outcomes in mice. These results suggest that targeting endogenous FA represents a promising therapeutic strategy for MS and other demyelinating disorders. Further investigations are warranted to explore the long-term safety, dosage optimization, and molecular pathways involved in FA-mediated neurotoxicity.

Chronic pain is a complex condition shaped by long-standing alterations in both physiological and psychological processes. Rather than representing a simple continuation of acute nociceptive signaling, it is increasingly understood as the outcome of progressive dysregulation within distributed neural systems that govern sensation, affect, motivation, and cognitive control. Neuroimaging and electrophysiological studies indicate that this state is accompanied by extensive plastic changes in deep brain structures and large-scale networks. Beyond well-described central sensitization processes, chronic pain is characterized by disrupted oscillatory rhythms and altered connectivity within large-scale brain networks, including thalamo-cortical circuits and prefrontal-limbic-reward networks. These findings support a conceptual shift from viewing chronic pain as a focal, lesion-driven phenomenon toward recognizing it as a disorder of distributed network pathology. Pharmacological treatments remain central to clinical practice, yet their long-term efficacy is often limited and frequently accompanied by substantial side effects. The ongoing concerns about opioid-related risks and the inadequate therapeutic response in a subset of patients highlight the need for safe, non-pharmacological approaches that can address not only pain but also comorbid disturbances in mood, sleep, and social functioning. Neuromodulation provides a promising path toward mechanism-based and non-pharmacological management of chronic pain by employing physical or chemical stimulation to alter the excitability and synchrony of specific neural populations within central, peripheral, and autonomic systems. While invasive deep brain stimulation demonstrates that targeting deep brain structures can be effective, its clinical application is restricted by surgical risks and cost, highlighting the importance of non-invasive techniques capable of reaching deep targets. Current non-invasive approaches, such as transcranial electric stimulation, are constrained by limited penetration depth and insufficient spatial precision. These limitations hinder reliable engagement of deep regions implicated in pain, including the thalamus and nucleus accumbens, and tend to produce broad, non-specific modulation of cross-network oscillatory activity. Temporal interference (TI) stimulation has emerged as a means of overcoming these obstacles. By delivering interacting high-frequency currents that generate a low-frequency envelope within the head, TI enables focal stimulation of deep targets while minimizing superficial current delivery. Recent multiscale modeling and animal studies indicate that TI exploits the nonlinear rectification properties of neuronal membranes in response to high-frequency carriers, as well as their phase-locked responses to low-frequency envelopes, to generate “peak-focused” electric fields in deep regions under relatively low superficial current loads. Moreover, TI appears to exhibit potential advantages in terms of cell-type selectivity and rhythm-specific engagement, including differential responses across neuronal subtypes and distinct coupling to θ-, β-, and γ-band oscillations. These features suggest a promising avenue for correcting abnormal rhythms and network dynamics that contribute to chronic pain. This review summarizes current knowledge of the neural mechanisms underlying chronic pain and recent advances in TI research. It examines functional disturbances across key pain-related regions and networks, outlines the principles and technical characteristics of TI, and discusses potential deep-brain targets and stimulation strategies relevant to chronic pain. Evidence to date indicates that TI, with its non-invasiveness, tolerability, and capacity for precise deep brain modulation, holds great promise for the management of treatment-resistant chronic pain and may evolve into a new generation of precise and efficient non-pharmacological analgesic strategies.

The Golgi body, a core organelle in eukaryotic cells, plays a critical role in protein modification, sorting, vesicular transport, and serves as a key site for lipid synthesis and glycosylation. Glucose and lipid metabolism are central processes for cellular energy maintenance and biosynthesis, and are closely linked to Golgi function. Recent studies have revealed the extensive involvement of the Golgi body in regulating glucose and lipid metabolism, where maintaining its structural and functional homeostasis is crucial for normal physiological activity. Under various stress conditions such as acidosis, hypoxia, and nutrient deficiency, the Golgi body undergoes structural and functional disruption, leading to Golgi stress. This in turn activates specific signaling pathways, such as those mediated by the cAMP-responsive element binding protein 3 (CREB3) and proteoglycans, to alleviate Golgi stress and enhance Golgi function. Golgi stress contributes to glucose and lipid metabolic disorders by affecting the activity of insulin receptors, glucose transporters, and lipid metabolism-related enzymes. For example, Golgi stress triggers the cleavage and release of the active fragment of CREB3, which enters the nucleus and upregulates the transcription of ADP-ribosylation factor 4 (ARF4) and key gluconeogenic enzymes, including phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase). ARF4 promotes vesicle retrograde transport between the Golgi and endoplasmic reticulum, maintains secretory capacity, and enhances hepatic glucose output. This pathway is particularly active under high-fat or lipotoxic stress, leading to fasting hyperglycemia. When damaged Golgi components accumulate beyond a tolerable threshold, the cell initiates an autophagic response, selectively encapsulating the damaged Golgi into autophagosomes, which then fuse with lysosomes to form autolysosomes, leading to Golgiophagy. This process results in the degradation and clearance of damaged Golgi, thereby regulating Golgi quantity, quality, and function. Golgiophagy also plays a significant role in regulating glucose and lipid metabolism. For instance, under high-glucose conditions, autophagic flux may be suppressed, impairing the timely clearance and renewal of damaged Golgi, compromising its normal function, and further exacerbating glucose metabolism disorders. Additionally, Golgiophagy may participate in lipid degradation and influence lipid synthesis and transport. Research indicates that Golgi stress and Golgiophagy play important roles in glucose and lipid metabolism-related diseases. For example, the leucine zipper protein (LZIP) under Golgi stress conditions can promote hepatic steatosis. In mouse primary cells and human tissues, LZIP induces the expression of apolipoprotein A-IV (APOA4), which increases peripheral free fatty acid uptake, resulting in lipid accumulation in the liver and contributing to the development of fatty liver disease. This review systematically outlines the structure and function of the Golgi apparatus, the molecular regulatory mechanisms of Golgi stress and Golgiophagy, and their synergistic roles. It further elaborates on how Golgi stress and Golgiophagy participate in the regulation of glucose and lipid metabolism, discusses their clinical significance in related diseases such as diabetes, fatty liver disease, and obesity, and highlights potential novel therapeutic strategies from the perspective of Golgi-targeted medicine. Finally, this article addresses the challenges and future directions in Golgi-targeted interventions, aiming to advance the clinical translation of such strategies and foster breakthroughs in the treatment of glucose and lipid metabolism-related disorders.

Objective The accuracy of Y-chromosome haplogroup assignment is crucial for tracing paternal lineage in male samples. With the advancement of high-throughput sequencing technologies, high-density Y-SNP genotyping from whole-genome or array-based data has become a standard method for determining Y-chromosome haplogroups. This study systematically evaluated the performance of 4 commonly used high-density SNP genotyping systems—namely, the Global Screening Array (GSA), Chinese Genotyping Array (CGA), Affymetrix array, and the 1240K capture panel—for haplogroup assignment. This work provides a reference for data comparison across different systems.Methods We extracted genotype data for the 4 Y-SNP panels from 30× whole-genome sequencing (WGS) data of 1 590 male samples from the 1000 Genomes Project. Additionally, GSA array genotype data from 384 relative pairs (spanning 1st- to 12th-degree relationships) from 109 Chinese Han families were collected. Haplogroup assignment was performed using Y-LineageTracker v1.3.0 software. We assessed the concordance and resolution of haplogroup assignments between the four Y-SNP panels and the WGS data. The consistency and resolution of haplogroup assignments were also evaluated for both the 1000 Genomes Project samples and the 109 family samples collected in this study. Furthermore, the impact of varying numbers of Y-SNPs on haplogroup assignment was examined.Results The GSA and CGA panels demonstrated superior resolution and discrimination of haplogroup subclades compared with the other two panels. The haplogroup assignments from the GSA, CGA, and 1240K panels showed high concordance with WGS data, with consistency rates exceeding 88.70%, whereas the Affymetrix platform exhibited a significantly lower consistency rate of 61.89%. Specifically, the GSA and CGA panels consistently demonstrated superior performance compared with the other two panels in the assignment of haplogroups O-M175 and H-L901, achieving complete concordance (100%) for both haplogroups. In contrast, the Affymetrix panel erroneously assigned all individuals belonging to haplogroup O-M175 to haplogroup K2-M526. Furthermore, its accuracy for haplogroup H-L901 was exceedingly low, at merely 1.41%. This poor performance was characterized by the misassignment of 98.59% of H-L901 samples—specifically, 1.41% to J-M304 and a predominant 97.18% to F-M89. For haplogroup R-M207, all four panels exhibited uniformly high levels of consistency, with concordance values exceeding 94.00%. Notably, for haplogroup E-M96, the 1240K and Affymetrix panels outperformed the GSA and CGA panels in terms of concordance, representing the first instance in which these two panels surpassed the latter. Conversely, for haplogroups J-M304, Q-M242, and I-M170, all 4 panels showed relatively elevated misclassification rates, with the Affymetrix array demonstrating the poorest overall performance. None of the four panels showed any discordant haplogroup assignments among the familial relative pairs analyzed. A positive correlation was observed between the number of Y-SNPs (ranging from 1 000 to 10 000) and classification consistency; however, classification consistency plateaued when the number of Y-SNPs exceeded 10 000. Furthermore, a random sampling analysis conducted on the GSA and CGA panels demonstrated that the haplogroup misclassification rate exhibited negligible fluctuation across the Y-SNP range of 500 to 1 000. Conversely, a marked enhancement in classification consistency was observed as the number of markers increased from 1 000 to 5 000, ultimately reaching a plateau within the interval of 5 000 to 8 000 markers.Conclusion These findings indicate that the GSA and CGA panels provide high resolution and concordance, delivering reliable Y-haplogroup assignment for forensic investigations.

Abstract　Objective Donkey hide is the sole legally designated raw material for the preparation of the traditional Chinese medicine Ejiao. The quality stability of donkey hide during preservation directly determines the efficacy and safety of Ejiao. This study focuses on the dynamic succession of microbial communities during the preservation of donkey hides from different origins, aiming to clarify the correlation between microbial biodiversity difference and the degradation profiles of hide collagen and critical biochemical components, thereby providing a theoretical foundation for developing targeted preservation strategies based on microbial regulation. Methods Donkey hides originating from four different regions were subjected to an accelerated microbial aging assay to simulate the spoilage process. The microbial community succession was analyzed using high-throughput sequence. Microstructure changes and pore structure characteristics were assessed by scanning electron microscopy and mercury intrusion porosimetry, respectively. Additionally, the content of major components, including lipids, proteins, and sugars were determined by biochemical methods. Results After 96 h of aging, the collagen fiber structure in Africa donkeys hides (ADH) exhibited significant degradation and collapse, followed by Xinjiang donkeys hides (XDH). Instead, the microstructure of Dong'e black donkeys hides (DDH) and Peru donkeys hides (PDH) remained relatively intact. The porosities of DDH, XDH, PDH, and ADH increased from 27.9%, 15.7%, 30.3%, and 46.2% to 36.5%, 52.6%, 42.8%, and 57.7%, respectively, during the aging process, which suggested that the originally compact fiber structure was disrupted by microbial aging. Fourier transform infrared spectrometer analysis revealed the amide bands in XDH exhibited relatively weak intensity, and no collagen amide I band was observed in ADH. Meanwhile, the lipid and protein contents decreased in all four types of donkey hides, indicating these components served as the primary nutrient sources for the growth of microorganism. Notably, the most severe collagen degradation was observed in XDH and ADH. A substantial increase was detected in the total soluble sugar in PDH aging solution and hydroxyproline in the ADH aging solution, respectively. These results indicated that donkey hides exhibit distinct patterns of structural degradation and nutrient utilization. Furthermore, the viable cells number of donkey hides increased sharply after 48 h of aging. Metagenomic analysis revealed that the relative abundance of Euryarchaeota in ADH, PDH and XDH declining from initial 97.73%, 93.19% and 30.1% to 1.43%, 0.79% and 0.02% after 96 h, respectively. Conversely, a significantly increase was observed in the abundance of Firmicutes, with a marked increase in ADH, peaking at 92.75%. Additionally, the abundance of Pseudomonadota in PDH increased from 0.10% to 87.84%, suggesting that Bacillota and Pseudomonadota may be key factors exacerbating donkey hide spoilage. Unlike the other three types of donkey hides, the dominant bacterial phylum in DDH shifted from Pseudomonadota to Bacteroidota, characterized by a substantial abundance increase of Bacteroidota from 0.13% to 44.22%. Conclusion Regional variation in origin significantly influence the microbial aging of donkey hides, leading to distinct patterns of structural deterioration and differential nutrient utilization. Therefore, implementing origin-specific preservation strategies, through the precisely controlling environmental factors to suppress harmful phyla such as Bacillota, is crucial for enhancing the storage quality of donkey hides.

Objective This study aimed to construct a cell co-culture microfluidic chip based on droplet microfluidic to investigate the influence of multicellular interactions in complex microenvironments on the sensitivity of anti-tumor drugs.Methods We constructed a droplet microfluidic chip consisting of 12 co-culture units, with each unit containing 4 microwells for holding cell droplets, enabling the co-culture of 4 types of cells. To evaluate whether the co-culture of multiple cell types can be achieved in the droplet microfluidic chip, as well as to observe and analyze the interactions between different cell types, we investigated the interaction between microenvironmental cells and tumor cells through cell co-culture experiments. To construct a stable co-culture system capable of evaluating drug sensitivity, we conducted diffusion experiments with blue ink and model drugs to investigate the ability of drugs to diffuse in the chip and be taken up by cells. To investigate the effect of the complex microenvironment on cellular drug sensitivity, we carried out cell co-culture experiments combined with drug treatment to explore the changes in the drug sensitivity of tumor cells in the presence of microenvironmental cells. To explore the reasons for drug resistance in tumor cells under co-culture conditions, we detected DNA double-strand break marker using immunofluorescence.Results Experiments on cell culture within droplets showed that cells in each droplet exhibited good proliferation ability and consistent cell status, laying a foundation for the co-culture of multiple kind of cells. Cell co-culture experiments showed that compared with the mono-culture group, the numbers of LoVo cells, HUVECs, and macrophages in the co-culture group increased significantly. This confirms that there are obvious interactions between cancer-associated fibroblasts (CAFs), endothelial cells (HUVECs), macrophages, and tumor cells in the microenvironment, which promotes cell proliferation in the co-culture chip. Experiments on blue ink diffusion showed that drugs could diffuse uniformly and effectively into wells in different directions within the chip. Experiments on the diffusion of doxorubicin (a model drug) demonstrated that identical cells in different wells exhibited consistent drug uptake capacity for the drug. Additionally, cell co-culture experiments combined with oxiliplatin treatment revealed that with the concentration of 80 μmol/L, the survival rate of LoVo cells cultured alone was only 25%, whereas it reached 96% under co-culture conditions. with the concentration of 160 μmol/L, the survival rate of LoVo cells cultured alone was merely 2%, while that under co-culture conditions was 50%. These results indicate that the complex microenvironment composed of CAFs, HUVECs, and macrophages significantly reduces the drug sensitivity of LoVo cells to oxaliplatin through intercellular interactions. The immunofluorescence results showed that the expression level of γH2AX in LoVo cells decreased under co-culture conditions.Conclusion Our study achieved co-culture of the main constituent cells of the tumor microenvironment and analysis of their drug sensitivity in a droplet microfluidic chip for the first time. The research found that crosstalk between different microenvironmental cells strongly affects the drug sensitivity of tumor cells to oxaliplatin, suggesting that targeting the interactions between tumor microenvironmental cells is an effective strategy to improve the efficacy of tumor therapy. Our study provides new methods and approaches for the efficacy evaluation of anti-tumor drugs and the screening of new drugs. In addition, the open structural design of the co-culture chip can be combined with various omics technologies to analyze the molecular characteristics of cells under co-culture and drug treatment conditions. This is expected to provide new methods and experimental evidence for elucidating the mechanisms of drug action and identifying novel drug targets in the context of the microenvironment.

Circular RNAs (circRNAs) represent a distinct group of RNA molecules produced through back-splicing of precursor mRNAs. Their covalently closed structure, which lacks both a 5′ cap and a poly(A) tail, renders them highly resistant to exonucleolytic degradation and contributes to their remarkable intracellular stability. Although circRNAs were historically viewed as noncoding transcripts, accumulating evidence indicates that certain circRNAs can undergo translation under appropriate molecular contexts. Two major modes of noncanonical translation have been described so far: initiation mediated by internal ribosome entry sites (IRESs) and translation triggered by N6-methyladenosine (m6A) modification. These findings have broadened the traditional definition of noncoding RNA biology and suggest that circRNAs may contribute previously unrecognized elements to the cellular proteome. Peptides generated from circRNAs have been increasingly implicated in cancer biology. Depending on their molecular functions, these peptides may enhance malignant phenotypes—such as uncontrolled proliferation, motility, invasion, epithelial-mesenchymal transition, metabolic alteration, or drug resistance—or, conversely, exhibit inhibitory effects on oncogenic pathways. Their dual and context-dependent functions highlight the complexity of circRNA-mediated regulation and suggest that these translation products participate in multiple layers of tumor initiation and progression. In this review, we synthesize current knowledge regarding the molecular mechanisms that enable circRNAs to be translated, with particular attention to IRES-driven initiation, m6A-dependent regulation, ribosome accessibility, and the structural determinants required for translation competence. We further summarize well-characterized circRNA-encoded peptides and discuss how they influence tumor-associated signaling networks. In addition, we examine the potential translational applications of these peptides, including their value as diagnostic indicators, prognostic markers, or therapeutic entry points. Their inherent sequence stability, relative expression specificity, and detectability in clinical specimens make circRNA-derived peptides promising candidates for future biomarker and therapeutic development. Overall, circRNA translation research is reshaping our understanding of RNA function and offers new perspectives for studying tumor biology. We propose that expanding investigations into circRNA-encoded peptides will not only improve the mechanistic resolution of cancer research but may also pave the way for innovative strategies in precision oncology, including RNA-based therapeutics and peptide-targeting interventions.

Diabetic Nephropathy (DN) is the leading cause of end-stage renal disease (ESRD) globally, representing a major global health burden with limited disease-modifying therapies. Podocyte injury serves as the core pathological hallmark of DN, and conventional treatments targeting metabolic disorders or hemodynamic abnormalities fail to reverse the progressive decline of renal function. Accumulating evidence over the past decade has established that high glucose-induced podocyte pyroptosis—a pro-inflammatory form of programmed cell death—is a key driving force in DN progression. Its core molecular mechanism hinges on the activation of the TXNIP-NLRP3 inflammasome axis. Under sustained hyperglycemic conditions, excessive reactive oxygen species (ROS) are generated via pathways including the polyol pathway, advanced glycation end products (AGEs) accumulation, and mitochondrial dysfunction. Concurrently, methylglyoxal (a glucose metabolite) mediates post-translational modification of thioredoxin-interacting protein (TXNIP). These events collectively trigger the dissociation of TXNIP from thioredoxin (TRX), a redox-regulating protein. The free TXNIP then translocates to the mitochondria, where it binds to The NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) and promotes inflammasome assembly. This assembly activates cysteine-aspartic acid protease 1 (caspase-1), which cleaves Gasdermin D (GSDMD) to generate its N-terminal fragment (GSDMD-NT). GSDMD-NT oligomerizes to form membrane pores, leading to podocyte swelling, rupture, and the release of pro-inflammatory cytokines interleukin-1β (IL-1β) and interleukin-18 (IL-18). These cytokines amplify local inflammatory responses, induce mesangial cell proliferation, and accelerate extracellular matrix deposition, ultimately exacerbating glomerulosclerosis. MCC950, a highly selective NLRP3 inhibitor, exerts its therapeutic effects through a multi-layered mechanism: it binds to the NACHT domain (NAIP, CIITA, HET-E and TP1 domain) of NLRP3 with nanomolar affinity, forming hydrogen bonds with key residues (Lys-42 and Asp-166) within the ATP-hydrolysis pocket to block ATP hydrolysis, thereby locking NLRP3 in an inactive conformational state. Additionally, MCC950 interferes with the protein-protein interaction between TXNIP and NLRP3 and regulates mitochondrial homeostasis to reduce ROS production. Preclinical studies have demonstrated that MCC950 dose-dependently reduces proteinuria, restores the expression of podocyte-specific markers (nephrin and Wilms tumor 1 protein, WT1), and alleviates podocyte foot process fusion and glomerulosclerosis in both streptozotocin (STZ)-induced type 1 diabetic models (characterized by absolute insulin deficiency) and db/db type 2 diabetic models (driven by insulin resistance). However, discrepancies in therapeutic outcomes exist across different models—some studies report exacerbated renal inflammation and fibrosis in STZ-induced models—which may stem from differences in disease pathogenesis, intervention timing (early vs. mid-stage disease), and dosing duration. Despite its promising preclinical efficacy, MCC950 faces significant translational challenges, including low oral bioavailability, insufficient podocyte targeting, potential hepatotoxicity, and drug-drug interactions with statins (commonly prescribed to diabetic patients for cardiovascular risk management). Furthermore, off-target effects such as the inhibition of carbonic anhydrase 2 have been identified, raising concerns about its safety profile. Nevertheless, its unique mechanism of action—directly blocking podocyte pyroptosis by targeting the TXNIP-NLRP3 axis—endows it with substantial translational value. In the future, strategies to overcome these barriers are expected to advance its clinical application: targeted delivery via nanocarriers (e.g., PLGA-PEG nanoparticles or nephrin antibody-conjugated systems) to enhance renal accumulation and podocyte specificity; precise patient stratification based on biomarkers such as serum IL-18 and renal TXNIP/NLRP3 expression to identify “inflammatory-phenotype” DN patients most likely to benefit; and combination therapy with sodium-glucose cotransporter 2 (SGLT2) inhibitors—whose metabolic benefits synergize with MCC950’s anti-inflammatory effects. These approaches hold great potential to break through clinical translation bottlenecks, offering a novel, precise anti-inflammatory treatment option for DN and addressing an unmet clinical need for therapies targeting the inflammatory underpinnings of the disease.

Cancer remains a leading cause of global mortality, necessitating the development of advanced therapeutic strategies with enhanced efficacy and reduced systemic toxicity. Among promising bioactive agents, lactoferrin (LF)—a multifunctional iron-binding glycoprotein abundantly found in mammalian milk and exocrine secretions—has garnered significant interest for its potent and multifaceted anti-cancer properties. This review provides a comprehensive analysis of the current understanding of LF"s role in oncology, encompassing its structural biology, diverse mechanisms of action, and groundbreaking advancements in its application through nano-engineering. LF exerts anti-tumor effects through multiple pathways, including extracellular action, intracellular action, and immune regulation. It demonstrates a remarkable affinity for cancer cell membranes, binding to overexpressed anionic components such as glycosaminoglycans and sialic acids, as well as to specific receptors including the Low-density Lipoprotein Receptor-related Protein 1 (LRP-1). This selective binding facilitates targeted uptake. Upon internalization, LF orchestrates a direct assault by inducing cell-cycle arrest in phases such as G0/G1 or S phase through the modulation of key regulators including cyclins, CDKs, and p53. Furthermore, it promotes programmed cell death via apoptotic pathways, involving caspase activation and downregulation of anti-apoptotic proteins such as survivin. A more recently elucidated mechanism is the induction of ferroptosis, an iron-dependent form of cell death characterized by overwhelming lipid peroxidation. Beyond direct cytotoxicity, LF acts as a potent immunomodulator. It enhances Natural Killer (NK) cell activity, modulates T-lymphocyte populations, and crucially reprograms Tumor-Associated Macrophages (TAMs) from a pro-tumor M2 state to an anti-tumor M1 state, thereby reversing the immunosuppressive tumor microenvironment (TME). The translation of LF"s potential has been significantly accelerated by nanotechnology. The inherent biocompatibility and natural tumor-targeting capabilities of LF make it an ideal platform for sophisticated drug-delivery systems. This review details various fabrication strategies for LF-based nanoparticles (NPs), including self-assembly, sol-oil emulsion, and electrostatic complexation, among others. Research demonstrates that nano-formulations not only protect LF from degradation but also enhance its bioactivity and anti-cancer potency. More importantly, LF NPs serve as versatile carriers for a wide array of therapeutic agents, including conventional chemotherapeutics, natural compounds, and imaging agents. These engineered systems enable synergistic therapy and facilitate site-specific delivery. Notably, the ability of LF to bind to receptors on the blood-brain barrier (BBB) has been leveraged to develop nano-systems for glioblastoma treatment. Other innovative designs utilize LF to modulate the TME—for instance, by alleviating tumor hypoxia to sensitize cells to radiotherapy and chemotherapy. Despite compelling pre-clinical evidence, the clinical translation of LF and its nano-formulations remains nascent. While early-phase trials have established a favorable safety profile for recombinant human LF, larger Phase III studies have yielded mixed results, underscoring the complexity of its action in humans. Key challenges include enhancing drug targeting, optimizing loading efficiency, ensuring batch-to-batch reproducibility, and achieving deep tumor penetration. Future research must focus on the rational design of next-generation LF-NPs. This entails developing standardized manufacturing protocols, engineering "smart" stimuli-responsive systems for targeted drug release in the TME, and constructing multi-targeting platforms. A concerted interdisciplinary effort is paramount to bridge the gap between bench and bedside. In conclusion, LF, particularly in its nano-engineered forms, represents a highly promising and versatile agent in the oncological arsenal, holding immense potential for precise and effective cancer therapy.

Atherosclerosis (AS), the primary pathological contributor to cardiovascular diseases (CVDs), has increasingly affected younger populations due to modern dietary habits and sedentary lifestyles. Current diagnostic modalities, including ultrasound, MRI, and CT, primarily identify advanced lesions and inadequately evaluate plaque vulnerability, thereby hindering early detection. Conventional treatments, which involve long-term medications associated with side effects such as hepatic injury and surgical interventions that carry risks of restenosis and hemorrhage, underscore the urgent need for non-invasive, cost-effective early diagnostic methods and targeted therapies. Gut microbiota metabolites are pivotal in AS pathogenesis, with trimethylamine N-oxide (TMAO) and short-chain fatty acids (SCFAs) serving as functionally opposing biomarkers. TMAO is produced when gut bacteria, specifically Firmicutes and Proteobacteria, metabolize dietary choline and carnitine into trimethylamine (TMA), which the liver subsequently converts to TMAO via flavin-containing monooxygenase 3 (FMO3); TMAO is then excreted in urine. Variability in TMAO levels is influenced by marine food consumption and FMO3 modulation, which can be affected by genetics, age, and diet. Mechanistically, TMAO exacerbates AS by disrupting cholesterol metabolism, inducing endothelial dysfunction through the elevation of reactive oxygen species (ROS) and pro-inflammatory cytokines such as IL-6, and reducing nitric oxide levels. Additionally, TMAO activates NF-κB and NLRP3 pathways while enhancing platelet reactivity. Clinically, elevated TMAO levels correlate with early AS and serve as predictors of mortality in patients with stable coronary artery disease (CAD) and acute coronary syndrome (ACS), as well as major adverse cardiovascular events (MACE) in stroke patients. Conversely, SCFAs—namely acetate, propionate, and butyrate—are produced by gut bacteria such as Akkermansia muciniphila and Faecalibacterium prausnitzii through the fermentation of dietary fiber. These metabolites exert anti-AS effects: acetate aids in maintaining metabolic homeostasis; propionate protects endothelial function and reduces plaque area; and butyrate fortifies intestinal barriers while suppressing inflammation. Furthermore, SCFAs cross-regulate bile acid metabolism, thereby influencing TMAO levels, and antagonize the pro-inflammatory and lipid-disrupting effects of TMAO. The use of TMAO and SCFAs as standalone biomarkers is constrained by limitations. TMAO lacks specificity, while SCFA levels fluctuate based on gut microbiota and dietary intake. Traditional AS risk assessment tools, which include clinical indicators, imaging techniques, and single biomarkers such as CRP, LDL-C, and ASCVD scores, overlook gut metabolism and demonstrate inadequate performance in younger populations. This review advocates for an “antagonistic-complementary” combined strategy: utilizing acetate and TMAO for early AS, propionate and TMAO for progressive AS, and butyrate and TMAO for advanced AS, addressing endothelial dysfunction, lipid deposition, and plaque stability/thrombosis risk, respectively. For clinical application, standardization of detection methods is crucial; liquid chromatography-mass spectrometry (LC-MS) is the gold standard, necessitating a unified sample pretreatment protocol, such as extraction with 1% formic acid in methanol. Additionally, dried blood spots (DBS) facilitate non-invasive testing, provided that dietary controls are implemented prior to detection, including a 12-hour fast and avoidance of high-choline and high-fiber foods. Existing challenges encompass the absence of standardized systems, limited large-scale validation, and ambiguous interactions with conditions such as hypertension. The authors’ team has previously established connections between gut metabolites and AS, including the reduction of TMAO as a preventive measure for AS, thereby reinforcing this proposed strategy. Future research should prioritize standardization, the development of machine learning-optimized models, validation of interventions, and the exploration of multi-omics-based “gut microbiota-metabolite-vascular” networks. In conclusion, the combined detection of TMAO and SCFAs offers a novel framework for AS risk assessment, facilitating early diagnosis and targeted interventions while enhancing the integration of gut metabolism into cardiovascular disease management.

Deciphering how the brain enables humans to interact, coordinate, and learn from one another remains one of the most compelling challenges in contemporary cognitive neuroscience. Social interaction is a dynamic, reciprocal process. Over the past decade, hyperscanning research has consistently identified inter-brain synchronization (IBS) as a neural signature accompanying successful cooperation, communication, joint attention, and social learning. However, the correlational nature of these findings leaves a critical question unresolved: does IBS cause better social interaction, or does it merely reflect it? While traditional hyperscanning paradigms are powerful in revealing inter-brain neural dynamics “in the wild”, they cannot on their own determine the direction of causality. This gap has motivated the emergence of Multibrain Stimulation (MBS)—a new generation of causal inference tools designed to actively manipulate neural coupling across individuals. MBS leverages non-invasive transcranial electrical stimulation (tES) to modulate neural activity simultaneously in two or more interacting brains. Unlike conventional tES applied to a single individual, MBS employs coordinated stimulation parameters, such as synchronized waveforms or matched frequencies, to directly perturb the neural mechanisms underlying social interaction. By providing an exogenous, precisely controlled intervention on IBS, MBS satisfies interventionist criteria for establishing causal relationships: researchers can test whether modifying inter-brain synchrony leads to predictable changes in behavior, communication, or shared understanding. This capability represents a fundamental methodological shift, transforming interpersonal neuroscience from a largely descriptive discipline into one capable of mechanistic inquiry. The biophysical underpinnings of MBS vary depending on the specific modality used. Transcranial alternating current stimulation (tACS) functions through cross-brain entrainment: when two individuals receive oscillatory currents matched in frequency and phase (e.g., theta-, beta-, or gamma-band stimulation), their endogenous neural rhythms tend to align with the exogenous signal and, consequently, with each other. This alignment effectively instantiates principles of the Communication Through Coherence (CTC) framework, which posits that coherent oscillations optimize information exchange by synchronizing periods of excitability across neural populations. Meanwhile, transcranial direct current stimulation (tDCS) exerts its influence by altering the excitability of targeted cortical regions in a polarity-dependent manner, thereby tuning the computational readiness of social-cognitive hubs such as the temporoparietal junction, superior temporal cortex, or inferior frontal gyrus. A growing body of empirical evidence demonstrates that such manipulations yield robust behavioral effects. In joint motor tasks, in-phase tACS enhances interpersonal coordination by aligning motor preparation dynamics, reducing temporal variability, and enabling individuals to anticipate each other’s actions more effectively. In communication and social learning contexts, MBS targeting high-order integrative regions promotes conceptual alignment, accelerates knowledge transfer, and supports more efficient encoding of shared representations. Notably, the effects of MBS often persist beyond the stimulation period, suggesting short-term plasticity in cross-brain networks. Post-stimulation improvements in synchronization and coordination indicate that MBS may temporarily recalibrate the neural architecture underlying social interaction. However, these benefits exhibit strong parameter specificity—precise phase relationships (e.g., 0° in-phase versus 180° anti-phase) and frequency matching are essential for generating reliable behavioral outcomes. Taken together, MBS represents a transformative step toward establishing the causal principles of human sociality and offers a new avenue for probing how multiple brains become functionally aligned during interaction.

This paper presents a comprehensive exploration of the IPDPS teaching concept—a framework built upon the 5 core principles of interdisciplinarity, practicality, diversity, process-oriented, and soul-forging—and its systematic implementation in the general education course “Biology in Daily Life” at Sun Yat-sen University. Developed over 8 years of iterative practice, this educational model is designed to address critical challenges in cultivating top innovative talents within higher education. It specifically targets the overcoming of disciplinary barriers, the disconnection between academia and industry, the limitations of one-way knowledge transmission, the rigidity of traditional evaluation systems, and the lack of value guidance. The curriculum is innovatively structured around four life-centric modules—birth, aging, illness, and food—which seamlessly integrate cutting-edge advancements in life sciences with interdisciplinary knowledge, making complex biological concepts accessible and relevant to students from diverse academic backgrounds. Pedagogically, the course employs a rich array of teaching methods to activate student engagement and foster higher-order thinking skills. These include case-based learning? driven by real-world problems, multi-sensory interactive experiences?, storytelling? to illustrate scientific discovery processes, and contrastive analysis? of ethical dilemmas in science. A cornerstone of the implementation is a multi-tiered practical teaching system, encompassing mandatory in-class experiments, optional social investigations, corporate visits, and face-to-face sessions with industry leaders. This structure ensures learning extends from the classroom to real-world societal and industrial contexts. A significant reform is the shift from a summative to a process-oriented evaluation system. This system diversifies assessment methods and incorporates multiple evaluators, including teacher assessment, self-assessment, and structured peer review using detailed rubrics. This approach aims to stimulate intrinsic motivation, foster a growth mindset, and provide a more holistic measurement of student development. Fundamentally, the course deeply integrates value-shaping elements? into its fabric. By incorporating themes of national identity, scientific spirit, bioethics, and cultural confidence through specific cases, the course forges students’ sense of social responsibility and ethical reasoning, ensuring their innovative capacities are guided by a strong moral compass. Assessment data from 2017 to 2024 demonstrates significant positive outcomes. Course satisfaction ratings have shown a remarkable increase, rising from 80.5% to 96.8%. Survey data from 245 students (2021-2024) indicates that the course effectively broadens interdisciplinary horizons, enhances independent thinking and problem-solving abilities, and successfully integrates knowledge acquisition with capacity building and value orientation. The course has successfully functioned as an “initial incubator and screening mechanism”?for identifying and nurturing talented individuals, with some students even shifting their academic focus to biology as a result. In conclusion, the “Biology in Daily Life” course, underpinned by the IPDPS framework, provides a replicable and scalable paradigm for educational innovation in cultivating elite innovators. It represents a successful model for achieving the organic unity of knowledge impartation, ability cultivation, and value shaping in higher education. Future work will focus on optimizing differentiated content design for diverse student backgrounds, deepening practical teaching experiences, and establishing long-term tracking mechanisms for learning outcomes.

Objective Pioneer transcription factors (PTFs) possess the unique ability to recognize and bind their target DNA sequences within compacted nucleosomal DNA, thereby initiating chromatin opening and gene expression. They play pivotal roles in fundamental biological processes such as embryonic development, cellular reprogramming, and tumorigenesis. The specific regulatory mechanism by which nucleosomal rotational positioning governs PTF-nucleosome interactions remains inadequately elucidated. This study aims to systematically investigate the role of the rotational orientation of motifs in PTF-nucleosome binding.Methods We employed a DNA deformation energy model to predict the rotational positioning of DNA on nucleosomes. We analyzed high-throughput in vitro data from the NCAP-SELEX assay, which profiles the binding landscapes of numerous transcription factors to nucleosomal DNA. For in vivo analysis, we integrated genome-wide binding data (ChIP-seq) and nucleosome positioning data (MNase-seq) for eight well-characterized pioneer factors (OCT4, SOX2, KLF4, GATA4, MYOD1, FOXA1, CEBPA, and ASCL1) in human cells. Binding motifs were classified as “TF-bound” if they overlapped with ChIP-seq peaks and “TF-unbound” otherwise. DNA bendability profiles and Fast Fourier Transform (FFT) analysis were used to assess rotational positioning patterns around these motif sites. This analytical framework was further applied to specific biological contexts, including cellular reprogramming from IMR90 fibroblasts to induced pluripotent stem cells (iPSCs) and the differentiation of human embryonic stem cells (hESCs) to human neuroectodermal cells (hNECs).Results Our in vitro analysis revealed a strong dependence of transcription factor binding on the rotational orientation of TF-binding motifs. For SOX7, the unbound motifs at specific enrichment peaks exhibited a rotational phase clearly opposite to that of the SOX7-bound motifs. Similarly, analysis of P53 binding sequences confirmed that successful binding in vitro correlated with model-predicted exposure of the DNA minor groove at the motif center, consistent with P53’s binding mode. Genome-wide in vivo analysis of the eight PTFs showed that their DNA binding motifs were generally associated with DNA sequences exhibiting significant 10-bp periodicity in bendability, suggesting an inherent potential for nucleosome association. Crucially, for most factors (except ASCL1), the average rotational positioning preferences were remarkably similar between TF-bound and TF-unbound motifs. This indicates that, at a global genomic level, rotational positioning is not the primary determinant dictating whether a nucleosomal motif is bound by its cognate PTF in vivo. This phenomenon persisted during cellular reprogramming (IMR90 to iPSC), where the rotational positioning of OSKM factor motifs bound versus unbound in nucleosomal regions showed no significant overall difference. Interestingly, during hESC differentiation to hNECs, SOX2 binding sites underwent comprehensive reprogramming. In hNECs, the rotational positioning of nucleosomal SOX2-bound motifs was significantly different and, unexpectedly, opposite to the general preference observed in hESCs and for unbound motifs in hNECs, suggesting a cell context-dependent rewiring of binding mechanisms.Conclusion This study suggests a distinction in the role of DNA rotational positioning in TF-nucleosome binding between in vitro and in vivo environments. While rotational positioning critically governs the binding efficiency of factors like SOX7 and P53 in simplified in vitro systems, PTFs in vivo appear to overcome this steric hindrance at the binding interface. The ability of PTFs to bind nucleosomal motifs, even when key interaction surfaces are partially buried, might stem from their unique structural properties (e.g., intrinsically disordered regions, DNA distortion/binding domains), nucleosome breathing which transiently exposes DNA, and potential cooperativity with other factors. Our results highlight the unique capacity of pioneer factors to drive chromatin openness through mechanisms beyond rotational positioning.

Methamphetamine (METH) addiction is a severe and increasingly prevalent neuropsychiatric disorder for which current diagnostic and therapeutic approaches remain limited and predominantly symptom-oriented. Exercise, as a safe, accessible and cost-effective non-pharmacological intervention, has emerged as a promising strategy to ameliorate METH-induced neurotoxicity and addiction-related behaviors. Growing evidence indicates that these benefits are closely linked to the regulation of exercise-induced biomarkers, defined as molecular indicators whose expression or activity is dynamically altered during or after physical activity. This review focuses on the core regulatory role of exercise-induced biomarkers in METH addiction and systematically summarizes their involvement in key neurobiological pathways, outlining molecular pathological mechanisms such as dysregulation of dopamine, glutamate and GABA neurotransmitter systems, neuroinflammation and oxidative stress, and epigenetic remodeling, and emphasizing how these processes converge on changes in candidate biomarkers in the brain and periphery. On this basis, the review describes how exercise modulates neural plasticity, neurotransmitter systems, inflammation and oxidative stress through biomarkers such as brain-derived neurotrophic factor (BDNF), exerkines, inflammatory cytokines, metabolites and non-coding RNAs, with particular attention to neurotrophic and immune-related markers, microRNAs and other epigenetic regulators that can reverse METH-induced synaptic and structural abnormalities and promote recovery of cognitive and emotional functions. Advances in high-throughput omics technologies, including transcriptomics, metabolomics and multi-omics integration, are summarized to illustrate the screening and identification of key exercise-responsive biomarkers. Studies in METH-addicted animal models have revealed differentially expressed genes, signaling pathways (e.g., PI3K-Akt, mTOR, Wnt) and core nodes such as NFKBIA and CXCL12 that may mediate the protective effects of exercise. The review further discusses the potential of exercise-mediated biomarkers as objective indicators for diagnosis, dynamic monitoring of therapeutic efficacy and patient stratification. Multi-gene diagnostic models based on peripheral samples (e.g., hair follicles, blood) demonstrate how biomarker panels can distinguish non-recovered, almost-recovered and healthy individuals, providing a molecular basis for staging METH use disorder and evaluating the impact of exercise interventions. The temporal dynamics of biomarker changes before and after exercise are highlighted, underscoring the value of longitudinal monitoring of factors such as BDNF, immune-related genes and circulating microRNAs to capture treatment-relevant windows of plasticity. In addition, the underlying molecular basis of exercise as an adjunct therapy and gene-targeted exercise strategies that leverage individual biomarker and gene expression profiles to optimize exercise prescriptions are summarized. Current conceptual and technical challenges are outlined, including heterogeneity of biomarker responses, individual variability, assay sensitivity and specificity, and gaps between preclinical findings and clinical application, together with future directions for integrating exercise with multi-omics, artificial intelligence-assisted biomarker discovery and, prospectively, gene-editing-based interventions. Particular emphasis is placed on the need to standardize exercise protocols, incorporate stage-specific and sex-sensitive designs, and combine exercise with pharmacotherapy and psychosocial rehabilitation in real-world clinical settings across diverse healthcare systems. Overall, this review aims to provide a comprehensive and integrated mechanistic framework and updated theoretical support for the application of exercise-mediated biomarkers in the diagnosis, therapeutic effect monitoring and personalized intervention of METH addiction, and to offer new and clinically relevant insights into the development of precision medicine strategies for substance use disorders.

Objective Osteoporosis is a progressive metabolic bone disorder characterized by reduced bone mass and microarchitectural deterioration, leading to increased skeletal fragility and susceptibility to fracture. Conventional diagnostic and risk-assessment approaches, such as dual-energy X-ray absorptiometry (DXA) and the FRAX? algorithm, remain limited because they rely primarily on bone mineral density (BMD) and a restricted set of clinical factors, failing to capture the multidimensional determinants of bone strength. This study aimed to develop and validate a deep learning-based multi-dimensional feature fusion model that integrates heterogeneous biological, structural, functional, and genetic information to improve the early identification of osteoporosis and enhance fracture risk prediction.Methods A total of 12 856 participants were aggregated from three major data repositories: the International Osteoporosis Foundation database, a clinical research database on osteoporosis, and a large-scale medical informatics dataset. A unified data-extraction protocol was applied to ensure cross-database harmonization, followed by quality control, variable standardization, and missing-data handling using multiple imputation by chained equations (MICE). A multimodal deep learning framework was constructed to integrate six categories of features: BMD measurements, quantitative bone microarchitecture parameters, bone turnover biomarkers, established clinical risk factors, osteoporosis-related genetic polymorphisms, and sensor-derived balance and gait metrics. A multi-task learning strategy was adopted to simultaneously predict osteoporosis status and 10-year fracture probability. Model training used five-fold cross-validation, and external validation was conducted in an independent clinical cohort. Model performance was benchmarked against DXA alone and the FRAX tool.Results In the internal test cohort, the proposed model achieved an AUC of 0.936 (95% CI: 0.927-0.945), with a sensitivity of 87.5% and a specificity of 91.2%, significantly outperforming DXA alone (AUC=0.889) and FRAX (AUC=0.842) (both P<0.05). External validation yielded an AUC of 0.918 (95% CI: 0.905-0.931) and demonstrated strong calibration (Brier score=0.087). SHAP analyses revealed that, beyond BMD, key predictors included trabecular separation, serum C-terminal telopeptide of type I collagen, balance-related metrics, gait speed, and specific SNPs within the RANKL and VDR loci. A simplified model incorporating only BMD, clinical features, and bone turnover markers preserved high accuracy (AUC=0.917), underscoring its feasibility for resource-limited clinical environments.Conclusion The deep learning-based multi-dimensional feature fusion model markedly enhances the precision and individualization of osteoporosis assessment compared with traditional tools. By integrating biological, structural, metabolic, genetic, and functional dimensions of bone health, the model provides a comprehensive representation of skeletal integrity and robustly improves both diagnostic accuracy and fracture risk prediction. Its strong generalizability across demographic subgroups highlights its clinical applicability. This work offers a promising direction for developing next-generation intelligent decision-support systems that may meaningfully improve osteoporosis screening, risk stratification, and preventive care.

Ischemic stroke (IS) accounts for approximately 80% of all stroke cases and is a leading cause of death and long-term disability worldwide. Its core pathological mechanism involves the interruption of cerebral blood flow, leading to neuronal cell death and ischemic tissue necrosis in the brain, which is associated with multiple molecular processes including apoptosis, inflammation, and oxidative stress. This review systematically discusses the classification of HDACs, the mechanisms of action of HDAC inhibitors, and their multiple effects in inhibiting cell apoptosis, regulating neuroinflammation, repairing the blood-brain barrier, and improving cognitive function following IS. HDACs function by removing acetyl groups from histone lysine residues, leading to chromatin condensation and gene silencing. The HDAC family is classified into four classes: class I (HDAC1, 2, 3, 8), class IIa (HDAC4, 5, 7, 9), class IIb (HDAC6, 10), and class IV (HDAC11), with class III being the NAD⁺-dependent sirtuins. Histone deacetylase inhibitors (HDACi) exert significant neuroprotective effects following ischemic stroke through a multi-target, multi-pathway synergistic mechanism. The core mechanisms include inhibition of neuronal apoptosis, regulation of neuroinflammation, protection of the blood-brain barrier (BBB), and improvement of cognitive impairments (PSCI). HDACi regulate gene expression epigenetically by upregulating genes such as p21/CIP1, leading to cell cycle arrest, while also modulating apoptosis-related proteins by inhibiting pro-apoptotic signaling pathways, thereby reducing neuronal cell death. In terms of neuroinflammation, HDACi suppress NF-κB and activate Nrf2 pathways, decreasing the release of pro-inflammatory cytokines and preventing the pro-inflammatory polarization of microglia and macrophages, thus modulating the inflammatory response. Regarding BBB protection, HDACi regulate the expression and restoration of tight junction proteins such as Occludin and Claudin-5, while inhibiting the release of destructive factors like MMP-9, alleviating vasogenic edema, and maintaining BBB integrity. Furthermore, HDACi promote the transcription of neurotrophic factors and synaptic-associated genes, enhancing neuroplasticity and repairing neuronal networks, ultimately improving cognitive functions. Therefore, HDACi demonstrate great potential as a multifaceted therapeutic strategy for ischemic stroke. HDAC inhibitors (HDACis) represent a powerful multi-target therapeutic approach that transcends the limitations of traditional thrombolytic therapies. HDACis represent a powerful multi-target therapeutic approach that transcends the limitations of traditional thrombolytic therapies, which are hampered by a narrow time window and risks of reperfusion injury. Histone acetylation is increased by HDACis, which relaxes chromatin and reactivates protective gene transcription. Their selectivity and chemical structure are used to classify them. Trichostatin A (TSA) and sodium butyrate (SB), a short-chain fatty acid, are examples of broad-spectrum inhibitors that are effective in lowering infarct volume and reducing neuroinflammation. More selective inhibitors, including tubastatin A (HDAC6-selective) and entinostat (class I-selective), may have fewer adverse effects while increasing efficacy. By suppressing apoptosis by modifying the p53, Bcl-2, and JNK pathways, reducing neuroinflammation by blocking NF-κB and NLRP3 activation, preserving the integrity of the blood-brain barrier by strengthening tight junction proteins, and promoting synaptic plasticity, neurogenesis, and the expression of neurotrophic factors like BDNF, these inhibitors provide neuroprotection through a variety of interrelated mechanisms.Despite their great potential, HDACis" clinical translation is fraught with difficulties, mostly because of non-selective inhibition-related adverse effects such as hepatotoxicity and gastrointestinal problems with valproic acid (VPA). In order to accomplish targeted delivery to the brain, future research is consequently shifting toward the development of highly selective inhibitors, refining dosing regimes, and utilizing cutting-edge drug delivery technologies like nanoparticles. In summary, the development of effective neuroprotective and neurorestorative treatments for IS may be greatly aided by a nuanced, spatiotemporally accurate understanding of HDAC activities and the judicious use of subtype-selective HDACis.

Parkinson’s disease (PD), the second most prevalent neurodegenerative disorder worldwide after Alzheimer’s disease, is pathologically characterized by the progressive loss of dopaminergic neurons in the substantia nigra pars compacta and the abnormal intracellular aggregation of α-synuclein into Lewy bodies. Traditionally, the clinical symptoms of PD have focused on motor dysfunction, which includes characteristic signs such as resting tremor, rigidity, bradykinesia, and postural instability. However, increasing evidence from both clinical and basic research suggests that the clinical presentation of PD is highly diverse, with neuropsychiatric complications representing a significant and unavoidable aspect of the disease"s overall burden. From the perspective of clinical phenotypes, the range of neuropsychiatric symptoms associated with PD is extensive, primarily including depressive disorders, generalized anxiety, apathy, impulse control disorders, and cognitive impairments related to executive function and memory. Notably, emotional and cognitive dysfunctions often manifest years prior to the onset of motor symptoms. This clinical observation indicates that the pathological processes of PD may originate within the non-motor circuits of the central nervous system (CNS), particularly in neural networks closely linked to emotional regulation and cognitive function. As one of the human body"s most lipid-rich organs, the CNS comprises lipids that account for approximately 50%-60% of the dry weight of brain tissue. These lipid molecules serve not only as structural components but also actively participate in the formation of cell membrane phospholipid bilayers, myelin sheath insulation layers, and various signal transduction complexes. From a functional perspective, lipids not only provide the structural foundation necessary for maintaining neuronal membrane fluidity, synaptic plasticity, and ion channel activity, but also act as essential molecules in energy metabolism, signal transduction, and epigenetic regulation. Notably, the frontal cortex—particularly its evolutionarily specialized prefrontal cortex (PFC)—functions as the brain"s "executive center for cognition and emotion". This region is pivotal for higher cognitive functions, including working memory, decision-making, and behavioral inhibition, as well as for the complex regulation of emotions, such as reward and risk assessment. This region displays an exceptionally high synaptic density and is abundant in structural lipids, including unsaturated fatty acids and cholesterol, which makes it particularly vulnerable to disturbances in lipid metabolism. In PD research, lipid imbalance has become a central focus. As investigations progress, the importance of lipid metabolic pathways becomes increasingly apparent. Simultaneously, pharmacological therapies aimed at lipid regulation show considerable efficacy in addressing cognitive and emotional deficits associated with PD. In light of this, the present study utilizes bioinformatics analysis to identify differentially expressed genes in the peripheral blood of PD patients, demonstrating significant enrichment in processes such as chronic depression, cholesterol metabolism, fatty acid metabolism, AMPK signaling pathways, and insulin resistance. Expanding on this groundwork, the present review systematically explores the connections between dysregulated lipid metabolism and metabolic reprogramming in cognitive and emotional impairments associated with PD. Through the analysis findings, intervention approaches focusing on various fundamental pathological pathways such as neuroinflammation, mitochondrial dysfunction, imbalance in lactate homeostasis, and disrupted protein homeostasis are suggested. These proposals provide innovative perspectives for advancing mechanistic investigations and therapeutic advancements targeting cognitive and emotional disorders in PD.

Spinal cord injury (SCI) is a highly disabling trauma of the central nervous system, characterized by a complex pathological process involving intertwined multiple mechanisms. Key pathological events include excessive activation of neuroinflammation, oxidative stress injury, neuronal apoptosis, autophagic dysfunction, and energy metabolism imbalance, which severely disrupt the integrity of spinal cord neural function and significantly reduce patients" quality of life. Currently, clinical neurorepair strategies for SCI have limited efficacy and are difficult to achieve synergistic intervention targeting multiple pathological links. Therefore, exploring novel core therapeutic targets and precise intervention regimens has become an urgent need in this field. The Sirtuins family (SIRT1-SIRT7), as NAD⁺-dependent deacetylases, play a central role in critical biological processes such as cellular metabolism regulation, immune homeostasis maintenance, and stress injury repair, and have been identified as potential intervention targets for neurological diseases. This review systematically summarizes the cellular localization and core biological functions of each member of the Sirtuins family, with a focus on their regulatory roles and molecular mechanisms in the pathological process of SCI: SIRT1, 3, 5, and 6 inhibit the excessive activation of the NF-κB pathway and block NLRP3 inflammasome assembly through deacetylation modification, thereby participating in the regulation of neuroinflammation after SCI; meanwhile, they alleviate oxidative stress injury in spinal cord tissues by activating the Nrf2 antioxidant pathway and enhancing the activity of antioxidant enzymes such as SOD and NADPH, forming a “anti-inflammatory-antioxidant” synergistic protective effect. SIRT7 delays neuronal apoptosis by promoting DNA damage repair and inhibiting apoptotic signaling pathways. SIRT3 and SIRT5 target mitochondrial function, improve mitochondrial energy metabolism by regulating the modification status of enzymes involved in the tricarboxylic acid cycle and oxidative phosphorylation, and restore autophagic homeostasis by modulating the acetylation levels of FOXO3a and AMPK, providing metabolic support for neural repair. We summarize that a variety of natural Chinese herbal components (e.g., resveratrol, matrine) and synthetic compounds (e.g., SRT1720, AGK2) can influence the pathological progression of SCI by targeting and regulating members of the Sirtuins family. We propose that Sirtuins-targeted combination therapeutic strategies (e.g., combined with stem cell transplantation, neurotrophic factor supplementation, or antioxidant intervention) are expected to break through the limitations of single therapies and enhance the repair effect of SCI through multi-mechanism synergistic actions. In conclusion, the Sirtuins family exhibits critical mechanisms of action and potential intervention value in the pathophysiological process of SCI. This review summarizes and prospects novel Sirtuins-targeted therapeutic strategies, aiming to provide new insights for basic research and clinical translation in this field.

As oncologic therapies continue to advance, the overall survival of cancer patients has markedly increased. Nevertheless, virtually every anticancer treatment modality is accompanied by some degree of cardiotoxicity. Epidemiological data indicate that approximately 30 % of cancer survivors ultimately die from cardiovascular disease. Among the cardiotoxic agents, the anthracycline doxorubicin (DOX) is the most widely used; it effectively suppresses a variety of malignant tumors—including breast cancer, lymphoma, and acute leukemia—but its cardiac toxicity limits further escalation of clinical dosing. Literature reports identify a cumulative dose of ≥250 mg/m2 as the threshold of high risk, with roughly 25 % of patients receiving DOX developing varying degrees of myocardial injury; severe cases progress to heart failure. Even at cumulative doses below the traditional safety limit, some patients exhibit cardiac dysfunction after the first administration, suggesting that cardiotoxicity is not solely a linear function of dose. DOX related cardiotoxicity can be classified as acute (hours to days after administration), sub acute (weeks to months), and chronic/late onset (years later). Most patients initially exhibit only mild reductions in left ventricular ejection fraction (LVEF) or subtle abnormalities in global longitudinal strain (GLS), often without symptoms. Recently, cardiac biomarkers (cTn, NT proBNP) combined with high sensitivity echocardiography (speckle tracking) have been recommended for monitoring high risk individuals, enabling detection of subclinical injury before overt LVEF decline. Currently, several preventive and therapeutic approaches are used in clinical practice, which can be summarized into the following four points: (1) dose limitation and administration strategies: fractionated low dose regimens, liposomal encapsulation, or continuous infusion lower peak plasma concentrations, thereby reducing cardiac exposure; (2) pharmacologic prophylaxis: β blockers (e.g., carvedilol) and ACE inhibitors/ARBs have shown protective effects on LVEF in some randomized trials, though results remain inconsistent and require larger confirmatory studies; (3) metabolic targeted interventions: animal experiments indicate that activation of PPARα or supplementation with L carnitine restores fatty acid oxidation and improves ATP generation, suggesting metabolic modulators as promising cardioprotective candidates; (4) lifestyle modifications: regular aerobic exercise up regulates mitochondrial biogenesis genes (PGC-1α) and reduces reactive oxygen species (ROS) production; small clinical studies have demonstrated a potential benefit in attenuating cTnT elevation. However, DOX-induced cardiotoxicity has not been effectively controlled, indicating that the core mechanism underlying DOX-related cardiac toxicity remains unidentified. Cardiomyocytes are high energy demand cells, and metabolic dysregulation is considered a central component of DOX induced cardiotoxicity. DOX disrupts myocardial metabolic balance through several interrelated pathways. (1) Oxidative stress and mitochondrial damage: DOX generates abundant ROS within cells, leading to mitochondrial membrane potential loss, lipid peroxidation, and iron accumulation, which suppress electron transport chain activity and markedly reduce ATP synthesis efficiency. (2) Autophagy dysregulation: DOX interferes with autophagic flux, preventing the clearance of damaged mitochondria and further aggravating apoptosis and inflammatory responses. (3) Inflammation and cytokine release: oxidative stress activates NF-κB, up-regulating pro inflammatory cytokines such as TNF-α and IL-6, creating a chronic inflammatory microenvironment that weakens myocardial contractility. (4) Epigenetic modifications: studies have shown that DOX alters DNA methylation and histone acetylation patterns in cardiomyocytes, affecting the expression of key metabolic genes (e.g., PGC-1α, CPT-1) and further inhibiting fatty acid β oxidation. These mechanisms collectively lead to suppressed fatty acid oxidation and compensatory up regulation of glycolysis, manifested by an elevated lactate/pyruvate ratio, accumulation of medium chain acyl carnitines, and a pronounced decline in ATP production. The resulting energy deficit precipitates left ventricular contractile dysfunction and, ultimately, heart failure. Despite extensive basic and clinical research on DOX cardiotoxicity, a unified risk assessment model and precise interventions targeting metabolic disturbances remain lacking. This review systematically summarizes recent progress on DOX induced cardiotoxicity and highlights that impairment of myocardial energy metabolism is a central mechanism of injury, thereby deepened our understanding of how impaired myocardial energy metabolism drives DOX induced injury, we can move toward safer chemotherapy protocols that achieve “cure cancer without harming the heart”.

The inflammatory response is the foundation and a critical component of innate immunity. It serves as a vital defense mechanism, enabling the body to rapidly recognize and resist the invasion of foreign pathogenic microorganisms through a spontaneous immune reaction. Through pattern recognition receptors (PRRs), the host can effectively identify pathogen-associated molecular patterns (PAMPs) from microbes like bacteria and viruses, as well as damage-associated molecular patterns (DAMPs) released by injured cells. This allows for swift identification and resistance against pathogenic invasions, fulfilling a cellular surveillance function. As one of the most important protein complexes in innate immunity, the NLRP3 inflammasome—a large multi-protein complex—is among the most extensively studied inflammasomes. It assembles in response to pathogenic invasion or other danger signals and is crucial for the processing and release of pro-inflammatory mediators. This process helps the body distinguish between “self” and “non-self” and plays a significant role in both inflammatory and antiviral responses, thereby maintaining the host’s internal homeostasis. However, under certain conditions, immune regulation can become dysregulated, leading to an inflammatory response that is either too weak or too strong. This imbalance between pro-inflammatory and anti-inflammatory states can ultimately result in disease and tissue damage. Notably, not all viral infections activate the inflammasome. The activation mechanism of the NLRP3 inflammasome remains unclear and is even a subject of debate. On one hand, viruses are recognized by the host’s innate immune system, which can activate the NLRP3 inflammasome to mobilize immune and inflammatory responses for antiviral defense. Upon viral infection, the host receptor protein NLRP3 recognizes inflammatory signals, recruits the adapter protein ASC, and forms an inflammasome complex with pro-caspase-1. This triggers a cascade of activation events that initiate the innate immune response. Strategies involved in this process include altering intracellular and extracellular ion concentrations, affecting host cell energy metabolism, and directly interacting with components of the NLRP3 inflammasome to regulate its activation. On the other hand, viruses have evolved multiple strategies to inhibit NLRP3 inflammasome activation and evade immune responses. These include regulating NLRP3 ubiquitination and degradation, inhibiting the assembly and activation of the NLRP3 inflammasome, and modulating its effector functions. Furthermore, while NLRP3 inflammasome activation upon viral infection helps clear the virus and is crucial for antiviral defense, viruses can also evade this immune mechanism to facilitate their own replication and proliferation. A deeper understanding of the interplay between inflammasome activation and viral replication will contribute to the precise and effective prevention and treatment of currently incurable viral diseases. Therefore, this article will focus on the complex interactions between viral infection and the NLRP3 inflammasome. It will review recent advances in understanding virus-induced NLRP3 inflammasome activation and the immune evasion strategies viruses employ by modulating NLRP3 inflammasome activity, with the ultimate goal of fundamentally controlling viral replication in the host. In-depth research in this area will not only enhance our understanding of viral pathogenesis but also provide new strategies for clinical antiviral therapy and drug development.

The alternative lengthening of telomeres (ALT) is a homology-directed repair (HDR)-based mechanism that maintains telomere length independently of telomerase by hijacking the canonical double-strand break (DSB) repair machinery. In ALT-positive cells, a RAD51-, MUS81-, and BLM-dependent recombination cascade copies telomeric tracts from sister chromatids, extrachromosomal telomeric circles (t-circles), or inter-chromosomal templates, thereby restoring a functional TTAGGG repeat array. This process is characterized by a distinct molecular signature:(1) chronic replication stress, manifested by elevated ATR-CHK1 signaling, R-loop accumulation, and fragile telomere phenotypes;(2) clustering of telomeric chromatin into ALT-associated PML bodies (APBs), which serve as SUMO-dependent recombination hubs enriched for SLX4-SLX1, MRE11-RAD50-NBS1, and FANCD2 complexes; and (3) global chromatin remodeling, marked by the eviction of histone H3.3 and its chaperones ATRX/DAXX, derepression of the long non-coding RNA TERRA, and acquisition of constitutive heterochromatin marks (H3K9me3/H4K20me3) along with the facultative heterochromatin mark H3K27me3. Together, these changes establish a chromatin environment permissive for homologous recombination. Importantly, these alterations are not merely passive by-products but are functionally required for homology search, strand invasion, and resolution of recombination intermediates. This is supported by CRISPR screens identifying ATRX, DAXX, and the SUMO E2 enzyme UBC9 as essential ALT fitness genes. While 85%-90% of human cancers re-express telomerase reverse transcriptase (TERT), the remaining 10%-15% are telomerase-null and rely exclusively on ALT for immortality. ALT tumors are enriched in osteosarcomas, glioblastomas, pancreatic neuroendocrine tumors, and aggressive soft-tissue sarcomas. In telomerase-negative somatic cells, progressive telomere shortening during each S phase eventually reaches a critical length, triggering a persistent DNA damage response (DDR) at chromosome ends. This activates the p53-p21 and p16INK4A-Rb tumor suppressor pathways, driving cells into stable replicative senescence. Although this telomere-length-dependent senescence acts as a potent barrier to malignant progression, recent single-cell analyses reveal that senescent fibroblasts and epithelial cells transiently display ALT-like features—such as accumulation of telomeric γH2AX/53BP1 foci, formation of APB-like PML condensates containing SUMOylated TRF1 and TRF2, and intermittent TERRA upregulation. These observations suggest that telomerase-negative tumors and senescent cells share a recombination-permissive chromatin state. Although senescent cells do not achieve net telomere elongation—likely due to intact p53/p16 checkpoints restraining unscheduled HDR—transient ALT activation may enable rare clonal escape. This further implies that ALT operates not only as a tumor-cell survival pathway but also as a protective mechanism against environmental stress. Indeed, spontaneous immortalization of TERT-/- fibroblasts in vitro is preceded by stochastic ALT induction, indicating that stochastic recombination at dysfunctional telomeres can overcome senescence barriers and initiate malignant transformation. Consistent with this model, whole-genome sequencing of ALT-positive tumors frequently identifies early driver mutations in TP53, ATRX, and DAXX, which disable replicative-senescence checkpoints while simultaneously enhancing telomeric HDR. Here, we synthesize the convergent molecular features of ALT tumors and senescent cells, highlighting:(1) replication stress as a common initiating cue, (2) SUMO-dependent phase separation as a platform for telomere-templated recombination, and (3) epigenetic erosion of ATRX/DAXX-mediated heterochromatin as a rate-limiting step. Finally, we discuss therapeutic implications: (1) pharmacological inhibition of SUMO E1/E2 enzymes to prevent APB scaffold nucleation, (2) synthetic-lethal exploitation of replication stress via ATR/CHK1 inhibitors, and (3) immune-microenvironment-targeting strategies that remodel the senescence-associated secretory phenotype (SASP). Collectively, this review elucidates the mechanisms by which ALT regulates cellular senescence and tumorigenesis, offering druggable vulnerabilities and translational strategies for the clinical management of telomerase-negative tumors.

Bispecific antibodies, engineered to simultaneously bind two distinct antigens or two epitopes on the same antigen, are now widely utilized in tumor therapy and various other fields. Depending on their mechanisms of action, bispecific antibodies can be designed into diverse structural formats, including IgG-like bispecific antibodies containing an Fc region. The Fc region mediates immune effector functions by interacting with receptors on immune cells or soluble immune components. However, antibodies containing an Fc region have a relatively high molecular weight, which limits their tissue penetration. They also exhibit slow systemic clearance in vivo and possess pharmacokinetic characteristics marked by a long terminal elimination half-life. Symmetric IgG-like bispecific antibodies feature a symmetric structure and are bivalent for each target antigen. During production, since the two heavy chains carrying the Fc region are identical, issues related to chain mispairing do not arise, thereby simplifying the manufacturing and purification processes. Moreover, the pairing of two identical natural Fc chains allows for correct disulfide bond formation, resulting in a more stable structure. Glycosylation of the Fc region remains in its natural state, preserving Fc-mediated functions. However, as the variable regions of the two antigen-binding sites are linked to the same heavy chain, the design must account for potential steric hindrance when the antibody binds both antigens simultaneously. In contrast, asymmetric IgG-like bispecific antibodies consist of two different heavy chains, each carrying antigen-binding domains that recognize distinct antigens or epitopes, offering greater structural design flexibility. Their development, however, requires addressing challenges related to heavy chain and light chain pairing. Strategies to prevent heavy chain mispairing include engineering the spatial configuration of the Fc region, facilitating Fab arm exchange, applying IgG-IgA chain exchange techniques, and introducing charge modifications in the Fc domain. To ensure correct light chain-heavy chain pairing, approaches such as introducing electrostatic interactions or novel disulfide bonds between the chains, swapping the CH1 and CL domains, or replacing the CH1-CL module with a T-cell receptor-derived structure have been employed. Non-IgG-like bispecific antibodies lack an Fc region. They are characterized by their small size and low molecular weight, which confer enhanced tissue penetration, rapid systemic clearance, and high structural versatility. Unlike IgG-based formats, they do not bind Fc receptors or activate the complement system directly. Different bispecific antibodies exert therapeutic effects through distinct mechanisms, which are largely determined by their structural design and target specificity. Currently recognized mechanisms of action include T cell redirection, dual signaling pathway blockade, immune checkpoint inhibition, formation of ternary complexes by binding two molecules, neutralization of soluble ligands, and acting as cofactors to mimic or enhance biological processes. Bispecific antibodies are extensively applied in cancer therapy. Beyond oncology, they are also being developed for the treatment of autoimmune diseases, infectious diseases, hematological disorders, and other conditions. Different structural designs offer unique advantages across therapeutic areas. This article elaborates on the structural designs of various types of bispecific antibodies and reviews their mechanisms of action and applications in therapeutics.

Viral membrane fusion proteins facilitate the fusion of viral and host cell membranes by undergoing a transition from a prefusion conformation to a post-fusion conformation, thereby enabling the transfer of viral nucleic acids into the cell interior. This transition process is characterized by peptide exposure, membrane insertion, and structural refolding. The prefusion configuration represents an optimal target for vaccine formulation and antiviral pharmacotherapy. However, the metastable nature of the prefusion conformation makes it prone to spontaneous conversion into the stable post-fusion conformation, thereby complicating structural analysis and vaccine design. Investigating the mechanisms of conformational change in these proteins and developing methods to stabilize their prefusion state remain challenging research topics. This review summarizes the structural and functional differences among three classes of membrane fusion proteins: class I proteins, which are predominantly composed of α-helices, form trimers, and rely on receptor binding or low pH to trigger fusion peptide release; class II proteins, which are mainly β-sheet-rich, rearrange from dimers to trimers and activate fusion loops via low pH; and class III proteins, which combine α-helices and β-structures, with mechanisms involving internal fusion loop insertion and membrane remodeling. It is evident that a comprehensive understanding of the mechanisms underlying viral membrane fusion is crucial for developing effective stabilization strategies for the prefusion conformation of these proteins. This paper presents several such methods that have been successfully employed in this endeavor, including: disulfide bond formation to stabilize domain-domain interactions; hydrophobic cavity filling to enhance core stability; proline substitution to restrict structural transitions in hinge regions; and multimer domains stabilizing the trimeric conformation. The stabilization strategies summarized and discussed herein have been validated in studies of multiple viral membrane fusion proteins and further applied in the design of vaccine antigens. Moreover, this paper highlights the potential applications of novel techniques, such as time-resolved cryo-EM, in capturing conformational intermediates and resolving dynamic transition processes. Such stabilization efforts, informed by structural insights, have yielded promising outcomes—for instance, prefusion-stabilized RSV F antigens that elicit potent neutralizing antibodies in clinical trials. Looking ahead, integrating computational modeling, such as AlphaFold predictions, with experimental data will further refine these approaches. Ultimately, these innovations promise to enable structure-guided therapeutics to combat emerging viral threats. This review provides a theoretical foundation for developing stable viral membrane fusion proteins, offering crucial insights for understanding viral membrane fusion mechanisms and advancing next-generation vaccines and antiviral drugs.

Chinese hamster ovary (CHO) cells are the most established and versatile mammalian expression system for the large-scale production of recombinant therapeutic proteins, owing to their genetic stability, adaptability to serum-free suspension culture, and ability to perform human-like post-translational modifications. More than 70% of biologics approved by the U.S. Food and Drug Administration rely on CHO-based production platforms, underscoring their central role in modern biopharmaceutical manufacturing. Despite these advantages, CHO systems continue to face three persistent bottlenecks that limit their potential for high-yield, reproducible, and cost-efficient production: excessive metabolic burden during high-density culture, heterogeneity of glycosylation patterns, and progressive loss of long-term expression stability. This review provides an integrated analysis of recent advances addressing these challenges and proposes a forward-looking framework for constructing intelligent and sustainable CHO cell factories. In terms of metabolic regulation, excessive lactate and ammonia accumulation disrupts energy balance and reduces recombinant protein synthesis efficiency. Optimization of culture parameters such as temperature, pH, dissolved oxygen, osmolarity, and glucose feeding can effectively alleviate metabolic stress, while supplementation with modulators including sodium butyrate, baicalein, and S-adenosylmethionine promotes specific productivity (qP) by modulating apoptosis and chromatin structure. Furthermore, genetic engineering strategies—such as overexpression of MPC1/2, HSP27, and SIRT6 or knockout of Bax, Apaf1, and IGF-1R—have demonstrated significant improvements in cell viability and product yield. The combination of multi-omics metabolic modeling with artificial intelligence (AI)-based prediction offers new opportunities for building self-regulating CHO systems capable of dynamic adaptation to environmental stress. Regarding glycosylation uniformity, which determines therapeutic efficacy and immunogenicity, gene editing-based glycoengineering (e.g., FUT8 knockdown or ST6Gal1 overexpression) has enabled the humanization of CHO glycan profiles, minimizing non-human sugar residues and enhancing drug stability. Process-level strategies such as galactose or manganese co-feeding and fine control of temperature or osmolarity further allow rational regulation of glycosyltransferase activity. Additionally, in vitro chemoenzymatic remodeling provides a complementary route to construct human-type glycans with defined structures, though industrial applications remain constrained by cost and scalability. The integration of model-driven process design and AI feedback control is expected to enable real-time prediction and correction of glycosylation deviations, ensuring batch-to-batch consistency in continuous biomanufacturing. Long-term expression stability, another critical challenge, is often impaired by promoter silencing, chromatin condensation, and random genomic integration. Molecular optimization—such as the use of improved promoters (CMV, EF-1α, or CHO endogenous promoters), Kozak and signal peptide refinement, and incorporation of chromatin-opening elements (UCOE, MAR, STAR)—helps maintain durable transcriptional activity, while site-specific integration systems including Cre/loxP, Flp/FRT, φC31, and CRISPR/Cas9 can enable single-copy, position-independent gene insertion at genomic safe-harbor loci, ensuring stable, predictable expression. Collectively, this review highlights a paradigm shift in CHO system optimization driven by the convergence of genome editing, synthetic biology, and artificial intelligence. The transition from empirical optimization to rational, data-driven design will facilitate the development of programmable CHO platforms capable of autonomous regulation of metabolic flux, glycosylation fidelity, and transcriptional activity. Such intelligent cell factories are expected to accelerate the transformation from laboratory-scale research to industrial-scale, high-consistency, and economically sustainable biopharmaceutical manufacturing, thereby supporting the next generation of efficient and customizable biologics manufacturing.

Alzheimer’s disease (AD) is a common chronic neurodegenerative disorder of the central nervous system characterized by progressive impairments in memory, cognition, and behavior, eventually leading to severe dementia and loss of self-care ability. Despite decades of investigation, the precise molecular mechanisms underlying AD remain incompletely understood, and effective disease-modifying treatments are still lacking. The traditional pathological hallmarks of AD including amyloid β-protein (Aβ) plaques and neurofibrillary tangles (NFTs) composed of hyperphosphorylated Tau fail to account for the complex biochemical and cellular alterations observed in AD brains. Ferroptosis, a distinct iron-dependent form of non-apoptotic programmed cell death, is increasingly recognized as a contributor to AD pathogenesis. Ferroptosis is driven by excessive accumulation of lipid peroxides and reactive oxygen species (ROS), leading to oxidative destruction of cellular membranes. Unlike apoptosis or necrosis, ferroptosis is morphologically characterized by shrunken mitochondria with condensed membrane densities and biochemically defined by the loss of glutathione peroxidase 4 (GPX4) activity. Disruption of iron homeostasis, a central hallmark of ferroptosis, triggers a cascade that inhibits the cystine/glutamate antiporter (System Xc^-), suppresses glutathione (GSH) synthesis, and impairs GPX4-mediated detoxification of lipid peroxides, leading to uncontrolled lipid peroxidation and oxidative stress that ultimately trigger ferroptotic cell death. This iron-driven cell death exhibits distinct morphological and biochemical characteristics compared with other forms of cell death. Ferroptosis contributes to AD pathogenesis through multiple mechanisms and is closely associated with disease onset and progression. Iron overload can affect early amyloid precursor protein processing, accelerate Aβ production and plaque deposition, reduce Tau protein solubility, and promote Tau hyperphosphorylation and aggregation into NFTs. Therapeutic strategies targeting ferroptosis—such as iron chelation with deferoxamine to reduce labile iron levels and inhibit Fenton reaction-driven oxidative damage; supplementation with antioxidants such as α-tocopherol or α-lipoic acid to neutralize reactive oxygen species (ROS) and scavenge lipid radicals; and administration of selenium or activators of the Nrf2-SLC7A11-GPX4 axis and the SIRT1/Nrf2 signaling pathway to restore glutathione-GPX4 function—can effectively block lipid peroxidation and suppress iron-dependent cell death. By modulating iron metabolism, enhancing antioxidant defenses, and inhibiting lipid peroxidation, these approaches hold promise for mitigating ferroptosis-related neuronal injury. These interventions collectively aim to modulate iron metabolism, strengthen antioxidant defenses, and suppress lipid peroxidation, thereby mitigating neuronal injury and delaying cognitive deterioration. Ferroptosis represents a pivotal intersection of iron metabolism, oxidative stress, and neurodegeneration in AD. Exploring ferroptotic mechanisms not only deepens our understanding of AD pathophysiology but also opens new avenues for therapeutic intervention. This review aims to comprehensively summarize the molecular basis of ferroptosis, elucidate its pathological roles in AD, and propose ferroptosis-centered therapeutic strategies, thereby providing a theoretical framework for future research and drug development in AD.

Objective With the continuous evolution of severe acute respiratory syndromes-coronary virus 2 (SARS-CoV-2) Omicron subvariants, particularly the emergence of BA.2.86 and its descendant JN.1, the efficacy of current neutralizing antibodies has faced substantial challenges. The JN.1 variant, noted for its pronounced immune evasion capacity, has rapidly become the globally dominant strain. Elucidating its escape mechanisms is therefore essential to guide the development of next-generation broad-spectrum vaccines and neutralizing antibody therapeutics. This study aimed to investigate the immune evasion mechanisms of JN.1 against broadly neutralizing antibodies, focusing on the effects of key receptor-binding domain (RBD) mutations on antibody binding and neutralization, thereby providing theoretical support for countering ongoing viral evolution.Methods We employed a multidisciplinary approach to systematically assess the binding and neutralizing activities of three broad-spectrum neutralizing antibodies (XGv074, XGv302, and XGv303) against BA.2.86 and JN.1. Binding affinities (K_D values) of antibodies to variant RBDs were determined using bio-layer interferometry (BLI). Cryo-electron microscopy (cryo-EM) was used to resolve the structure of the BA.2.86 Spike trimer in complex with antibody antigen-binding fragments (Fabs), achieving a resolution of 3.47 ? for the BA.2.86 S-trimer bound to XGv302. Molecular dynamics simulations and binding free-energy decomposition were conducted to quantify the contributions of key mutations at the antibody-RBD interface. Additionally, sequence alignment and structural modeling were performed to evaluate the role of conformational flexibility in the antibody heavy-chain complementarity-determining region 3 (HCDR3) in mediating tolerance to mutations.Results Experimental data showed that XGv074, XGv302, and XGv303 retained neutralizing activity against BA.2.86 but exhibited markedly reduced binding to JN.1, with only XGv074 maintaining weak neutralization (IC₅₀=2.3 mg/L). Cryo-EM structures revealed that all three antibodies targeted the RBD tip, overlapping with the ACE2-binding region. The JN.1-specific L455S mutation disrupted the hydrophobic interaction network between XGv302 and the RBD (involving key residues such as Y421 and L455), resulting in complete loss of neutralization. Binding free-energy decomposition further identified L455 and Y421 as energetic hotspots (ΔG<-3 kcal/mol), with the L455S mutation directly impairing antibody binding. XGv074, owing to greater conformational flexibility in its HCDR3 region, partially tolerated the mutation and retained weak binding. Molecular dynamics simulations showed that the L455S mutation not only eliminated the energetic contribution of this residue but also caused a concurrent decrease in binding free energy of neighboring residues, thereby reducing overall interface stability.Conclusion The JN.1 variant escapes broad-spectrum neutralizing antibodies primarily through the L455S mutation in the RBD, which disrupts energetic hotspots and remodels the antibody-binding interface. Antibody conformational flexibility enhances adaptability to such mutations, providing new insights for broad-spectrum antibody design. These findings highlight the critical roles of epitope energy distribution and antibody flexibility in maintaining neutralization breadth, offering essential guidance for the rational design of next-generation vaccines and antibody therapeutics: specifically targeting conserved energetic hotspots while enhancing CDR flexibility to counter immune evasion driven by viral evolution.