Abstract: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.

Abstract: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.

Abstract: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 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.

Abstract: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.

Abstract: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. 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.

Abstract: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 combined 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.

Abstract: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.

Abstract: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.

Abstract: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.

Abstract: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.

Abstract: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”.

Abstract: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 mass, 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 mass, 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.

Abstract: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.

Abstract: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.

Abstract: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. 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.

Abstract: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.

Abstract: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.