Decellularized extracellular matrix (dECM) leverages native architecture and bioactive components for tissue regeneration, yet its therapeutic efficacy is constrained by donor tissue maturity. While mature-tissue-derived dECM (Mat-dECM) lacks developmental signals, developmental-stage dECM (Dev-dECM) retains these cues but is limited by scarce sources and poor adaptability to adult environments. Here, we proposed postnatal maxillofacial odontogenic tissues as a novel Dev-dECM (pDev-dECM) source. Leveraging its unique trans-stage development—spanning embryonic crown formation to postnatal periodontal maturation—pDev-dECM balances developmental potency with adult environment adaptability. Multiomics comparative analysis revealed that pDev-dECM is enriched in arginylglycylaspartic-acid-containing proteins, which activate macrophage integrin αvβ5-mediated efferocytosis. This mechanism drives macrophage polarization toward regenerative M2 phenotypes, ensures adaptability to the adult environment, and reprograms mesenchymal stem cells to orchestrate developmental recapitulation. Consequently, pDev-dECM not only enabled in situ hierarchical regeneration of the periodontal complex but also facilitated cross-organ hierarchical regeneration in skin and muscle defect models. These findings demonstrate that pDev-dECM exerts spatiotemporal control over developmental recapitulation, establishing a universal biomaterial paradigm for multiscale tissue reconfiguration.

Mitochondrial DNA (mtDNA) damage and its subsequent release into the cytoplasm are strongly linked to osteoarthritis (OA), but the pathogenic mechanism remains poorly understood. Here, this study reveals that under inflammatory or oxidative stress, the down-regulation of mitochondrial base excision repair enzyme 8-oxoguanine DNA glycosylase 1 and excessive opening of the mitochondrial permeability transition pore jointly drive mtDNA escape into the cytoplasm. Activation of 8-oxoguanine DNA glycosylase 1 with TH10785 reduces the production of oxidized mtDNA and preserves mtDNA integrity, while suppression of excessive mitochondrial permeability transition pore opening with cyclosporin A prevents mtDNA translocation. The combined intervention synergistically decreases cytosolic mtDNA levels, alleviating cartilage matrix degradation and cellular senescence. Mechanistically, cytosolic mtDNA induces the senescence-associated secretory phenotype by activating the cyclic guanosine monophosphate-adenosine monophosphate synthase–stimulator of interferon genes–nuclear factor κB signaling axis, whereas combined intervention blocks this cascade activation. Notably, intra-articular injection of the combination of TH10785 and cyclosporin A markedly reduces senescence and ameliorates the progression of the experimental OA model mice. This research reveals the dual regulatory roles of mtDNA integrity and translocation in governing cytosolic mtDNA content, providing novel insights for developing mtDNA-targeted therapeutic strategies against OA.

In single-cell microbes, primary and secondary metabolism are synergistically integrated to support survival and environmental adaptation through the biosynthesis of structurally diverse and biologically active small molecules—known as secondary metabolites (SMs) or natural products, such as penicillin. However, the therapeutic potential of microbial SMs is often limited because their biosynthetic gene clusters remain silent or weakly expressed under laboratory conditions. In this study, we employed BioNavi-NP as a guide to identify heme A synthase (ClHAS) as a novel metabolic link in Curvularia lunata, bridging primary and secondary metabolism. Surprisingly, inactivation of ClHAS led to the diversification of its characteristic indolizidine alkaloids, yielding previously unreported molecular frameworks. Mechanistically, ClHAS disruption impaired heme biosynthesis, thereby inducing oxidative stress that activated an uncharacterized basic-region leucine zipper transcription factor (A7370). A7370 binds specifically to a unique cis-element (ACGGCTGAC) in the promoter region of cuaF, a pathway-specific positive regulator of indolizidine alkaloid biosynthesis. This cascade regulation produces structurally unprecedented alkaloids, some of which show antibacterial activity equal or superior to the clinically prescribed drug, tinidazole, against a range of human pathogenic bacteria. Overall, this work reveals a conserved regulatory network that coordinates primary and secondary metabolism in fungi, providing new mechanistic insights into the complexity of fungal SM biosynthesis.

Diabetes hinders nerve recovery after spinal cord injury (SCI). The complex pathological factors of diabetes increase the difficulty of treating diabetes combined with SCI. Maintaining normal microglial function is essential for SCI recovery. However, it is unclear whether diabetes hinders nerve recovery after SCI by influencing normal microglial function. This study explored the role and regulatory mechanism of diabetes in microglial function during SCI recovery. We constructed a type 2 diabetes (T2D) combined with SCI mouse model and confirmed that T2D hinders nerve repair after SCI. T2D blocked phagocytizing function of microglia in SCI mice, which results in increased myelin debris accumulation and poor remyelination. A mechanistic study demonstrated that T2D triggers activation of NLRP3 inflammasome by activating the RAGE–ROS–TXNIP axis and then induces excessive microglial pyroptosis, which consequently leads to considerable loss of microglia after SCI. Verapamil (VRP; a TXNIP inhibitor) treatment confirmed that TXNIP is necessary for NLRP3 inflammasome activation. Conditional microglial Caspase-1 gene knockout (KO) mice also confirmed that excessive microglial pyroptosis is an important inducing event for more severe nerve damage in T2D combined with SCI mice. Moreover, this T2D effect on increased microglial pyroptosis was also effectively reversed by N-acetyl-l-cysteine (NAC; an antioxidant) and N-benzyl-4-chloro-N-cyclohexylbenzamide (FPS-ZM1; a RAGE inhibitor). In conclusion, this study revealed that T2D induces increased microglial pyroptosis by activating the RAGE–ROS–NLRP3 axis, and then blocks remyelination after SCI, which strongly suggests that microglial pyroptosis may be the key target for treating T2D combined with SCI.

The development of fast-charging anode materials is crucial for overcoming the sluggish reaction kinetics of conventional graphite electrodes in lithium-ion batteries. This work presents a kilogram-scale producible hard carbon/graphene flake (HC-GF) composite anode engineered based on an “electron Velcro” conductive network design. In this architecture, highly conductive GFs are intimately integrated with the HC matrix, creating multidimensional electron/ion transport channels while effectively minimizing interfacial resistance. The resulting HC-GF anode exhibits exceptional electrochemical performance, delivering a reversible capacity of 506.4 mAh g⁻¹ and maintaining 223.8 mAh g⁻¹ at a 10 C rate (corresponding to 6-min fast charging). Remarkably, when blended with graphite at a 20% ratio in a 5 Ah LFP//graphite + 20% (HC-GF) pouch cell, this minimal doping level substantially enhances multiple performance metrics, particularly fast-charging capability and cycling stability. The pouch cell retains 3.39 Ah after 600 cycles at 3 C, corresponding to an impressive capacity retention of 94.6%. This study provides a valuable material strategy for advancing fast-charging lithium-ion battery technologies.

A comprehensive understanding of the physiological functions of mammary glands and the mechanisms underlying breast diseases is crucial for promoting women’s health and well-being. This review systematically summarizes the development and applications of in vitro mammary gland models, focusing on key components including cell types, hydrogel matrix, and breast-on-a-chip platforms. We begin with a detailed overview of primary mammary epithelial cells, various cell lines, organoids, and multicellular coculture systems, highlighting their respective characteristics and advantages in mimicking mammary gland physiology and pathology. Next, we examine the essential role of hydrogel matrix in providing extracellular matrix support and facilitating cellular interactions. Furthermore, recent advances in breast-on-a-chip technologies are reviewed, covering their design, fabrication, and functional capabilities that enable high-throughput drug screening and disease mechanism investigations. Finally, we explore the applications of these in vitro models in mammary gland development, breast cancer research, and drug discovery, underscoring their critical role in improving prevention and treatment strategies. By integrating these state-of-the-art technologies, this review provides researchers with a comprehensive perspective to better understand and apply in vitro mammary models in innovative studies.

Mitochondrial dysfunction is pivotal in the pathogenesis of cardiac ischemia/reperfusion (I/R) injury. Restoring mitochondrial function represents a promising strategy for mitigating I/R-induced cardiac injury. Mitochondrial fission regulator 1-like protein (MTFR1L), a recently identified mitochondrial dynamics protein, is abundantly expressed in the cardiac tissues. However, its functional role in I/R injury remains undefined. Here, Mtfr1l-knockout mice and human embryonic-stem-cell-derived cardiomyocytes are utilized to investigate the role of MTFR1L in myocardial I/R injury and elucidate its contribution to mitochondrial integrity and function. MTFR1L deficiency markedly worsened I/R-induced cardiac injury and mitochondrial dysfunction. These phenotypes were partially reversed by mitochondria-anchored apoptosis-inducing factor (AIF) overexpression. Mechanistically, MTFR1L protects the heart via 2 interconnected pathways. First, MTFR1L sustains AIF dimerization and stabilizes the AIF–CHCHD4 (coiled-coil-helix-coiled-coil-helix domain containing 4) complex, thereby preserving mitochondrial contact site and cristae organizing system integrity and cristae architecture to facilitate electron transport chain supercomplex assembly, sustain mitochondrial respiration, and limit reactive oxygen species production. Second, by physically interacting with AIF, MTFR1L prevents its mitochondrial release and nuclear translocation, thereby suppressing intrinsic apoptosis. Overall, these findings identify MTFR1L as a cardioprotective protein against myocardial I/R injury through a dual mechanism, providing new insights into the functional repertoire of MTFR1L beyond its previously recognized role in mitochondrial dynamics. Targeting MTFR1L or its interactors may offer novel therapeutic strategies for alleviating mitochondrial dysfunction and myocardial injury.

The rise of multidrug-resistant bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA), poses a serious threat to human health, necessitating an urgent need to develop innovative antibacterials with multifunctions for this. Here, focusing on integrating multi-antimicrobial mechanisms with visual monitoring, we report a novel phenanthro[9,10-d]imidazole fluorogen 9b, outstanding for its antibacterial effects against S. aureus and various clinical MRSA isolates (minimum inhibitory concentration = 0.5 to 1 μg/ml) with rapid bactericidal activity, high membrane selectivity, and low susceptibility to drug resistance. Further investigation revealed its dual antimicrobial mechanisms of action by concurrently disrupting bacterial cell membranes and inducing bacterial DNA degradation to accelerate bacterial death. Ultraviolet–visible and fluorescence spectroscopy studies showed that the fluorescence intensity of 9b was dramatically enhanced when it interacted with S. aureus. Notably, in vivo studies demonstrated that 9b effectively treated both skin and thigh MRSA infections, showing a favorable safety profile and superior efficacy compared to the first-line antibiotic vancomycin. The “dual-mechanism” fluorescent phenanthro[9,10-d]imidazole fluorogen 9b holds promise as a candidate for developing innovative antibacterial agents that enable simultaneous real-time diagnosis and treatment.

Recent advances in transcranial ultrasound stimulation (TUS) pulsed at 40 Hz have demonstrated the potential to ameliorate cognitive deficits in mouse models of Alzheimer’s disease. However, technical barriers remain as general anesthesia is required for mice, which restricts the accurate elucidation of biological mechanisms and behavioral effects under awake physiological conditions. Here, we report a wearable, free-moving ultrasound stimulation system that delivers TUS pulsed at 40 Hz to female 5xFAD transgenic mice to systematically evaluate the behavioral outcomes and underlying mechanistic pathways. Among the treatment groups, a 14-d regimen at an acoustic intensity of 2.14 W/cm² yielded the optimal cognitive outcome in Alzheimer’s disease mice, which was consistently verified across Y-maze and Morris water maze tests. Additionally, this group showed reduced Aβ plaque deposition and increased plaque-associated microglial activity. Furthermore, enhanced gamma oscillations in the hippocampus were detected following treatment. RNA sequencing revealed modulation of innate immune and inflammatory pathways. Corresponding molecular analysis demonstrated a marked down-regulation in RIPK1, phosphorylated NF-κB, and necroptosis markers, alongside reductions in key pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α). Collectively, our findings suggest that the cognitive improvement observed after treatment with TUS pulsed at 40 Hz may be linked to the modulation of neuroinflammatory and necroptotic pathways, possibly involving RIPK1/NF-κB signaling.

The dynamic metaverse paradigm integrates emerging technologies and offers transformative opportunities to enhance consumer healthcare applications through immersive, connected experiences. However, this paradigm faces substantial cybersecurity challenges, such as distributed denial-of-service attacks, probing, and port scanning. This undermines the trustworthiness and resilience of healthcare analytics frameworks. To address these threats, intrusion detection systems that support proactive anomaly detection are essential for securing metaverse-based healthcare applications. Conventional anomaly detection techniques face challenges such as low interpretability, suboptimal feature selection, class imbalance, and inefficient hyperparameter tuning. These challenges limit their reliability in practical cyber threat intelligence settings. To solve these challenges, this paper presents an anomaly-detection framework for Internet of Things-enabled metaverse healthcare environments. The proposed framework leverages an off-policy proximal policy optimization (PPO) algorithm that incorporates SHapley Additive exPlanations-based feature selection and class-specific reward adjustments to address imbalance. The reinforcement learning-based off-policy PPO enables adaptive, sample-efficient learning by leveraging prior experience during policy updates. The hyperparameters of the model are optimized using the Bayesian Optimization Hyperband algorithm to accelerate training and enhance performance. This optimization technique combines Bayesian search with the Hyperband method to improve efficiency and convergence during model tuning. The performance of our model is evaluated on NSL-KDD, MAWI, and CICIoT2023 datasets. The results depict that the model outperformed its contemporaries with state-of-the-art results where accuracy, F-measure, G-means, and area under the curve reached 88.005%, 87.271%, 87.986%, and 0.870; 92.184%, 88.992%, 89.738%, and 0.873; and 89.368%, 88.312%, 89.039%, and 0.836, respectively. The results confirm the effectiveness of the framework in cyber threat scenarios. They also show their potential for explainable, trustworthy intelligence in metaverse healthcare.