ABSTRACT Cisplatin is a widely utilized chemotherapeutic agent for the treatment of malignant cancers. However, its clinical application is significantly restricted due to its severe ototoxicity. Therefore, identifying protective medications to prevent cisplatin‐induced hearing loss (CIHL) is of paramount importance. Here, we show that AC591, derived from the classical herbal formula “Huangqi Guizhi Wuwu decoction,” exhibits notable protective effects on cisplatin‐induced ototoxicity. Our findings indicate that pretreatment with AC591 mitigates cisplatin‐induced auditory dysfunction, with its mechanism of action linked to the inhibition of p53‐mediated ferroptosis. By identifying the active components of AC591 in serum and cochlear tissues following oral administration, we recognized 18β‐glycyrrhetinic acid as an effective constituent for hearing protection. Furthermore, we confirmed that 18β‐glycyrrhetinic acid exerted otoprotective effects by binding to p53 and regulating proteins associated with ferroptosis. Collectively, our findings suggest that AC591 and 18β‐glycyrrhetinic acid may serve as promising agents for preventing CIHL.
ABSTRACT An organ‐on‐a‐chip platform has emerged as an advanced in vitro model that replicates organ‐level structures within physiologically relevant human microenvironments. In this review, we focus on intervertebral disc (IVD) degeneration, a leading cause of chronic low back pain, providing a comprehensive overview of the current understanding of IVD anatomy, physiology, and pathogenesis. We review conventional in vitro, ex vivo, and in vivo models, which have expanded our understanding; however, they inadequately reproduce the human phenotype, heterogeneous disc composition and anatomy, which comprises nucleus pulposus (NP) and annulus fibrosus (AF) with neovascularization and sensory innervation within the three‐dimensional (3D) degenerative microenvironment. We emphasize a disc‐on‐a‐chip platform to address these limitations, which integrates microfluidics and biomimetic ECM‐based hydrogels within a controlled microenvironment to miniaturize the disc. It offers a promising platform to simulate the pathological disc milieu by incorporating gradient control of key signaling molecules implicated in disc degeneration. These include a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) and matrix metalloproteinase (MMP)‐mediated extracellular matrix (ECM) degradation; tumor necrosis factor alpha (TNF‐α) and interleukin‐1 beta (IL‐1β)‐driven inflammation via nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB); vascular endothelial growth factor (VEGF)‐promoted angiogenesis; and nerve growth factor (NGF)‐ and brain‐derived neurotrophic factor (BDNF)‐mediated sensory innervation and sensitization, thus contributing to nociception. Collectively, disc‐on‐a‐chip offers a next‐generation in vitro model for investigating mechanisms relating to ECM degradation, inflammation, innervation, vascularization, and nociception underlying IVD degeneration, enabling precision preclinical therapeutic evaluations.
ABSTRACT Chronic diabetic foot ulcers represent a persistent clinical challenge characterized by a “locked” inflammatory phase, biofilm‐mediated infection, and impaired tissue regeneration. Because conventional antibiotic and debridement therapies fail to resolve dysbiosis or stimulate healing, microbial antagonism has emerged as a potent biological principle for wound restoration. This narrative review integrates ecological evidence and mechanistic insights from animal models and early human studies to explore how beneficial microorganisms, including probiotics, bacteriophages, and competitive consortia, actively reshape diabetic wound environments. We examined the multifaceted mechanisms of these interactions, ranging from direct pathogen inhibition to host immunomodulation and metabolic signaling via conserved pathways such as the p40/epidermal growth factor receptor (EGFR)/PI3K axis. To address significant translational barriers, we introduce a “Regenerative Microbiology” framework that emphasizes spatiotemporal coordination and precision stratification. By tailoring the use of live biotherapeutics or metabolically independent postbiotics to a patient's vascular and microbial profiles, this approach offers a strategic roadmap for transforming the management of infection‐driven tissue damage in chronic diseases.
ABSTRACT Post‐traumatic stress disorder (PTSD) is a debilitating psychiatric disorder affecting approximately 3.9% of individuals worldwide over their lifetimes. Since adolescence is characterized by ongoing brain maturation and increased responsiveness to environmental stressors, this developmental stage may provide an important window for PTSD intervention. Growing evidence supports an association between ω‐3 polyunsaturated fatty acids (PUFAs), especially docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), and the restoration of stress‐induced neural damage. However, how ω‐3 PUFA supplementation during adolescence affects adult susceptibility to PTSD remains poorly understood. Here, we observed significantly reduced DHA and EPA concentrations in the prefrontal cortex (PFC) and serum of PTSD‐susceptible rats. Providing 1.2% DHA and EPA in the adolescent diet significantly reduced PTSD‐like phenotypes in adulthood, including excessive fear memory retention, depressive‐like and anxiety‐like behaviors, while also reducing serum levels of proinflammatory cytokines. These phenotypic and inflammatory improvements were associated with changes in the gut microbiome, including its composition and diversity. DHA/EPA supplementation was also linked to alterations in multiple lipid species, with serum phosphatidylcholine levels showing a significant correlation with fear memory expression. Moreover, we observed dysregulation of the PI3K/AKT/mTOR pathway in the PFC of PTSD‐like rats, which was less pronounced in DHA/EPA‐supplemented animals. Collectively, these findings provide the first evidence relating adolescent ω‐3 PUFA status to PTSD‐like phenotypes in adulthood. It further identifies that gut microbiota composition, lipid metabolic profiles (notably phosphatidylcholine), and prefrontal PI3K/AKT/mTOR signaling are correlated with these behavioral outcomes, suggesting that these systems may collectively contribute to trauma resilience.
ABSTRACT Degenerative ocular diseases are characterized by a convergence of molecular perturbations, including oxidative stress, chronic inflammation, failure of proteostasis, mitochondrial dysfunction, abnormal angiogenic signaling, and biomechanical remodeling. The eye's unique architecture, sealed by the blood‐retina barrier, presents formidable anatomical and cellular topological challenges to achieving long‐term curative therapeutics. Nanotechnology offers an advantage in potential therapy: by utilizing carriers in the nanometer size range and leveraging on appropriate surface modifications, it can effectively overcome these physiological barriers for enhanced, controlled, and targeted treatment. Clinical and preclinical systems such as antioxidant nanoparticles, sustained release implants, anti‐angiogenic nanoplatforms, multifunctional nanocomposites, and regenerative scaffolds illustrate how nanotechnology can overcome limitations in bioavailability, tissue penetration, and pathway specificity in degenerative ocular diseases. Diagnostic advances that include plasmonic contrast agents, quantum dots, graphene biosensors, smart contact lenses, and photothermal neuromodulators further demonstrate continuous molecular surveillance and responsive intervention. Emerging materials such as DNA origami and magnetoelectric nanosystems expand this capability by enabling direct modulation of transcriptional, metabolic, and neuroelectrical pathways. Together, these developments position nanotechnology as a powerful strategy for addressing the molecular origins of ocular degeneration while also highlighting ongoing challenges in long term safety, device stability, manufacturing reproducibility, and regulatory standardization.
ABSTRACT Meta‐analysis is fundamental to evidence‐based medicine, yet traditional workflows remain labor‐intensive and susceptible to bias. Although LLM‐based research agents offer opportunities for workflow automation, they often lack the data fidelity and methodological traceability required for rigorous quantitative evidence synthesis, particularly when parsing multimodal scientific charts. To address this challenge, we introduce MacAma, a semi‐automated multi‐agent framework for protocol‐constrained and human‐verifiable meta‐analysis. MacAma operationalizes selected PRISMA 2020 reporting items, PICOS‐based eligibility logic, and SYRCLE risk‐of‐bias domains as structured prompts, decision rules, output fields, and audit records. Critically, MacAma adopts a risk‐aware automation strategy: Lower risk, repetitive, and protocol‐driven tasks, such as literature screening and drafting, are delegated to AI agents, whereas high‐impact steps that directly affect effect‐size estimation and statistical conclusions, such as quantitative chart‐data extraction, remain subject to expert verification. In a preclinical radiotherapy case study evaluating tumor‐related immune outcomes and metastatic potential mediated by circulating tumor cells, MacAma achieved competitive screening performance in the evaluated benchmark and reduced the manual screening burden by over 80% within the current workflow. The case study further demonstrates how structured agent outputs, predefined criteria, and audit records can support transparent screening, data extraction, statistical synthesis, and manuscript drafting. These results suggest that MacAma may provide a scalable and auditable framework for AI‐assisted meta‐analysis, although important limitations remain in full‐text access, quantitative chart data extraction, and expert interpretation of heterogeneity. MacAma is open‐source and available at https://github.com/YilinYuan/MacAma.
ABSTRACT The reprogramming of tumor‐associated macrophages (TAMs) from a pro‐tumoral M2 to an anti‐tumoral M1 phenotype is an attractive therapeutic strategy whose clinical translation is undermined by the systemic toxicity of currently available pharmacological approaches. Here, we demonstrate that non‐invasive and localizable pulsed electromagnetic fields (PEMFs) induce macrophage reprogramming downstream of transient receptor potential canonical 1 (TRPC1) channel activation. Brief (10 min) PEMF exposure polarized macrophages toward an M1 phenotype by activating Stimulator of Interferon Genes (STING)‐dependent NF‐κB inflammatory pathways that were abolished by TRPC1 knockdown or inhibition. PEMF exposure directly enhanced the immunogenicity of breast cancer cells and modified macrophage‐cancer crosstalk to promote M1 macrophage polarization and the attraction of STING‐activated macrophages to the cancer cells. In co‐cultures, PEMF exposure stimulated macrophage‐mediated phagocytosis of cancer cells in a STING‐ and TRPC1‐dependent manner. In spheroids, PEMFs induced the reprogramming of TAMs to an M1 status and selectively enhanced infiltration of M1 macrophages, resulting in STING‐mediated phagocytosis of cancer cells. In mice, 2 weeks of twice‐weekly PEMF exposure resorbed engrafted tumors and selectively eliminated cancer cells within tumors while promoting immune cell recruitment. PEMFs offer a non‐invasive manner to locally reprogram TAMs within the tumor microenvironment to preferentially eliminate cancer cells.
ABSTRACT Adipose‐derived stem cells (ADSCs) are central regulators of adipose tissue homeostasis and regenerative capacity. Accumulating evidence indicates that aging and obesity profoundly impair ADSC function, through progressive mitochondrial dysfunction and disrupted mitochondrial–nuclear communication. Emerging studies reveal that defects in nuclear–mitochondrial crosstalk constitute a key driver of ADSC senescence and adipose tissue aging. In this review, we synthesize recent advances in understanding the mitochondrial mechanisms underlying ADSC aging, with particular emphasis on how mitochondrial dysfunction reshapes stem cell fate decisions, metabolic plasticity, and inflammatory signaling within aged adipose niches. We further highlight mitochondria targeting therapeutic strategies that hold promise for reversing ADSC senescence. Collectively, this framework positions mitochondrial regulation as a unifying axis for ADSC rejuvenation, offering new opportunities to restore adipose tissue homeostasis and mitigate age‐related metabolic dysfunction.
Synergy therapy of Chinese herbs is an effective strategy for wound repair and its further development focuses on the chronological release of specific herbs with healing proceeding. Herein, we report a novel asiatic acid/baicalein nanocarrier integrated microcapsule with spatiotemporal release feature from microfluidic electrospray for wound healing. Benefiting from advantages of both nanocarrier formulation and integration capacity of microfluidic electrospray, synthesized baicalein-tannic acid nanoparticles are integrated into the shell of microcapsule while fabricated asiatic acid liposomes are located at the core region. This design enables chronological release: baicalein is first released to kill bacteria, followed by releasing asiatic acid liposomes to enhance cell migration and granulation tissue formation. In vitro tests confirm the excellent biocompatibility, antibacterial and pro-migration property of microcapsules. The results from in vivo wound healing studies showed the outcomes of reduced inflammation and accelerated wound closure in microcapsules-treated wounds. Therefore, it is believed that this herbal microcapsule with spatiotemporal and hierarchical release of baicalein and asiatic acid is an effective therapeutic platform for clinical wound treatment.
The molecular mechanisms linking premature and physiological aging remain incompletely understood. Here, we present a cross-species, multi-organ metabolic atlas of progeroid laminopathy (PL) generated through integrated lipidomic and metabolomic profiling of heart, lung, skin, and serum from Hutchinson-Gilford progeria syndrome (HGPS) mouse models and human PL cohorts. We identify organ-specific metabolic alterations, with the most pronounced changes observed in the skin at the omics level. Notably, we uncover disruption of a taurine-TCA-TUDCA cytoprotective axis, suggesting a coordinated metabolic vulnerability in PL. Serum profiling further reveals candidate circulating biomarkers, including increased LPC(18:1e) and LysoPA along with decreased D-glucose 1-phosphate and NAD. These alterations are conserved across species and, importantly, display concordant trends in independent cohorts of physiologically aged healthy individuals. This overlap indicates a partial convergence between PL-associated and normal aging-related metabolic changes. Together, our findings provide a metabolic framework for understanding PL pathophysiology and highlight clinically accessible biomarkers that warrant future longitudinal validation.
Liquid metal (LM)-derived electronic skin (e-skin) exhibits significant potential in the domains of human-machine interfaces, health monitoring and energy management. Herein, we introduce a cost-effective paradigm of LMs-derived e-skin featuring anti-freezing capability, efficient thermal management, and motion detection. The inexpensive and eco-friendly porous pomelo pith (PP) is employed as the substrate for the patterning of LM and as the physical scaffold for adhesive hydrogels. The unique porous architecture of the PP provides a natural template for the integration of LM and hydrogels, which facilitates the creation of electronic devices with improved flexibility and adhesion. The diverse patterning of nickel (Ni)-doped LM on the rough and porous pomelo pith surface is facilitated by the attractive force of a magnetic field applied below. The adhesive pre-gel is readily drawn into the pores of the pomelo pith due to capillary action. The resulting LM/PP hydrogel electronics are capable of rapid and stable Joule heating behavior and reliable motion detection. Additionally, the incorporation of glycerol endows this e-skin with superior freezing and desiccation resistance. Thus, this facile fabrication strategy successfully integrates excellent conductivity, adhesion, and environmental resilience, offering a promising paradigm for advanced wearable electronics.
ABSTRACT Tissue engineering holds immense promise to revolutionize regenerative medicine by enabling the fabrication of functional, patient‐specific tissues and organs for clinical translation, yet it continues to face persistent challenges in designing scaffolds that simultaneously recapitulate native tissue architecture, support cell viability, and enable efficient mass transport. Traditional fabrication has moved the field forward, yet routinely falls short of producing hierarchical, anisotropic, biomimetic structures under gentle conditions. Ice‐templating (or freeze‐casting), which is based on freeze‐induced microphase separation, reframes the problem as crystal‐growth engineering. This review summarizes current fabrication strategies and their underlying mechanism of ice‐templating technology from physical and chemical perspectives. We then highlight recent advances in ice‐templating for tissue engineering application fields such as 3D cell culture, wound healing, bone regeneration, nerve repair, and liver support, emphasizing the relationship between microstructure and biomedical functional performance. Finally, we discuss the key challenges in translating ice‐templated biomaterials from laboratory research to clinical practice and outline future directions to fully harness this versatile biomedical strategy.
Neuroinflammation is a core pathological mechanism in neurodegenerative diseases. Although natural many compounds, derived from traditional Chinese medicine have shown promise in modulating neuroinflammation, conventional evaluation methods remain inefficient and fail to meet modern drug development needs. This study aimed to develop a neuroinflammation-on-a-chip for efficient and accurate evaluation of the anti-neuroinflammatory activity of such compounds. By integrating gelatin methacryloyl (GelMA) hydrogel with a microchamber array structure into a multi-channel concentration-gradient microfluidic chip, we constructed a functional neuroinflammation-on-a-chip suitable for high-throughput drug screening. Preliminary results demonstrated that the chip can successfully model lipopolysaccharide (LPS)-induced neuroinflammation and test the anti-inflammatory effects of curcumin (Cur) and resveratrol (RSV). Relative to traditional approaches, the chip offers the advantages of low sample consumption, rapid detection, and high data reliability. This study provides a novel tool for the efficient evaluation of anti-neuroinflammatory activity of traditional Chinese medicine active compounds and offers an innovative platform for research on neuroinflammation-related diseases.
Although chemotherapy-induced bone loss is well-recognized during breast cancer treatment, the underlying mechanism remains to be further elucidated, especially in patients with obesity. In this study, the objective was to investigate the impact of genomic silencing and pharmacological inhibition of S1P synthesis on bone loss in doxorubicin-induced obese breast cancer mice. In vitro study, upon the treatment of doxorubicin combined with palmitic acid, the S1P generated by 4T1 cells was significantly increased, resulting in an increase in osteoclastogenesis by activating the S1PR1/p-STAT3/NFATc-1 pathway in bone marrow-derived macrophages. In vivo study, pharmacological intervention with Sphingosine kinases (SPHK) antagonist SKI II or biological inhibition with SPHK1 and SPHK2 short hairpin RNA significantly reduced S1P production and rescued the obese breast cancer-bearing mice from doxorubicin-induced bone loss, manifested by the decreased osteoclastogenesis and recovered bone microarchitecture. Similarly, the administration of the S1PR1 antagonist FTY720 also alleviated bone loss in the breast cancer-bearing mice fed a high-fat diet. These studies indicate that genetic silencing and pharmacological inhibition can suppress S1P-dependent bone loss in doxorubicin-induced obese breast cancer mice. S1P shows promise as a potential drug target for preventing chemotherapy-induced bone loss in patients.
Long non-coding RNAs (lncRNAs) constitute a critical class of regulatory molecules involved in cancer biology and play pivotal roles in tumor initiation and progression. Nevertheless, the biological functions of many newly identified lncRNAs in non-small cell lung cancer (NSCLC), as well as their potential therapeutic relevance, remain insufficiently characterized. In this study, high-throughput sequencing analysis of paired NSCLC tumor tissues and adjacent non-tumorous samples revealed that LINC00973 is significantly upregulated in tumor specimens. Moreover, elevated LINC00973 expression was found to be closely associated with poor clinical outcomes in patients with NSCLC. Functional assays showed that LINC00973 knockdown inhibits NSCLC cell proliferation, migration, and invasion while inducing apoptosis, whereas overexpression produces opposite effects. These observations were confirmed in vivo, where LINC00973 depletion markedly suppressed tumor growth and metastasis. Mechanistically, LINC00973 interacts with and stabilizes deltex E3 ubiquitin ligase 3L (DTX3L), preventing its ubiquitination-mediated degradation and activating the AKT signaling pathway. Therapeutically, RGD-modified exosome-mediated delivery of LINC00973 siRNA significantly inhibited NSCLC progression in mouse models. Moreover, a synthetic biology-based strategy enabling hepatic production of exosomes carrying LINC00973-targeting siRNA achieved robust anti-tumor effects. Together, these findings establish LINC00973 as an oncogenic lncRNA that promotes NSCLC progression via DTX3L stabilization and highlight LINC00973 as a promising therapeutic target.
Skeletal muscle is essential for voluntary movement and exhibits a remarkable capacity for regeneration following injury. NFIX, a member of the Nuclear Factor I (NFI) family of transcription factors, plays a critical role in both skeletal muscle development and regeneration. Despite its emerging importance, the molecular basis of NFIX-mediated DNA recognition and transcriptional regulation in skeletal muscle remains poorly defined. Here, we demonstrate that NFIX promotes key cellular processes in skeletal muscle cells, as siRNA-mediated knockdown of NFIX significantly reduces cell proliferation, increases apoptosis, and impairs differentiation. Transcriptomic analysis revealed that NFIX regulates a network of genes involved in muscle metabolism, stress responses, and immune inflammatory responses. Biophysical characterization showed that NFIX exists as a monomer in solution and binds palindromic DNA with a 1:1 stoichiometry. A high-resolution crystal structure of the NFIXDBD bound to palindromic DNA reveals a monomeric binding mode driven by base-specific recognition of the TGGCA motif. Mutations that disrupt key DNA-contacting residues abolished both DNA binding and transcriptional activation in luciferase reporter assays. Together, these findings define the molecular mechanism of NFIX-dependent gene regulation in skeletal muscle and establish a structural framework for its function, providing new insights into the potential therapeutic targeting of NFIX in muscle diseases.
Accurate assessment of sperm concentration and motility is critical for the diagnosis and management of male infertility. However, current methods, manual hemocytometer counting and commercial computer-aided sperm analysis (CASA) systems, are limited by labor intensity, human error, and variable performance under diverse sample conditions. Here, we present an artificial intelligence (AI)-driven computer vision tool for high-resolution, quantitative analysis of sperm motility and concentration. In a prospective study of 26 semen samples (22 patients, 4 donors), we benchmarked the AI model against manual tracking (using Fiji software) and a commercial CASA system (Hamilton Thorne IVOS II). Our method computed concentration and motility parameters, including straight-line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), linearity (LIN), amplitude of lateral head displacement (ALHmax), and beat cross frequency (BCF). Calibration using donor samples enabled accurate mapping of tracked sperm counts to concentrations. The AI tool presented a strong linear correlation with manual tracking (R 2 = 0.93-0.98; Root Mean Square Error (RMSE) = 3.3-7.3 μm/s for VSL, VCL, VAP), and outperformed CASA in both accuracy and consistency across all motility parameters. Post-calibration, ALHmax and BCF estimates improved substantially, with a 30%-50% reduction in RMSE. Grading of sperm motility by the AI model aligned closely with manual classification, avoiding the systematic misclassification typically observed with CASA. Furthermore, the AI system exhibited higher repeatability and robustness across duplicate samples and variable imaging conditions, with deviations below ± 2%. These findings demonstrate that our AI-based tool offers a quantitative and reliable alternative to current semen analysis platforms, supporting improved fertility diagnostics and potentially a more informative treatment process.
Human T-lymphotropic virus type 1 (HTLV-1)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a chronic neuroinflammatory disease. Given the established role of the gut-brain axis in other neurological diseases such as multiple sclerosis, the role of the gut microbiome in the pathogenesis of HAM/TSP remains a critical unexplored area. The aim of this study was to characterize alterations in the gut microbiome associated with HTLV-1 infection and its clinical stages. We performed a cross-sectional analysis of the gut microbiome from 112 Brazilian individuals, including 24 healthy controls and 88 HTLV-1-infected individuals at different disease stages: 38 HAM patients, 17 patients with intermediate syndromes, and 33 asymptomatic carriers. Fecal samples were collected and analyzed using Illumina MiSeq sequencing to assess bacterial composition and diversity. Functional analysis was performed to identify differentially enriched gene categories and Kyoto Encyclopedia of Genes and Genomes (KEGG) modules. Significant dysbiosis was observed in HTLV-1-infected individuals, characterized by reduced bacterial diversity, an inverted Firmicutes/Bacteroidetes ratio, and specific changes in bacterial genera. Notably, HAM patients exhibited decreased Faecalibacterium and increased Ruminococcus_g2 abundance. These associations should be interpreted with caution, as patient cohorts were significantly older and differed in sex distribution from healthy controls. Functional analysis revealed 13 differentially enriched gene categories and five KEGG modules that were more abundant in HAM patients, indicating alterations in metabolic processes. These findings provide the first comprehensive insight into gut microbiome changes associated with HTLV-1 infection and disease progression. This study provides the first comprehensive insight into gut microbiome changes associated with HTLV-1 infection and disease progression. The identified microbial signatures and functional alterations highlight potential diagnostic and therapeutic targets for HTLV-1-associated diseases, particularly HAM. These findings open new avenues for further research and clinical applications.
ABSTRACT Peripheral nerve injury (PNI) presents a significant clinical challenge due to the intrinsic limitations of nerve regeneration and poor functional recovery. Although nerve guidance conduits (NGCs) offer a promising alternative to autografts, their therapeutic efficacy is often constrained by insufficient bioactivity and electrical conductivity. To address these dual deficiencies, we engineered an electroactive living nerve conduit by integrating silk sericin (SS)‐modified carbon nanotubes (SCNTs) with adipose‐derived stem cells (ADSCs). The SCNTs serve as a conductive scaffold, whereas the ADSCs provide a sustained release of neurotrophic factors. This design creates a synergistic microenvironment to promote neuronal maturation and axonal regeneration. In an experimental rat model featuring a 10‐mm sciatic nerve gap, ADSC/SCNT/RAM NGCs demonstrated regenerative performance comparable to autografts, facilitating axon connection and recovery of motor functions. Histological assessment revealed that the implanted ADSC/SCNT/RAM NGCs promoted the most extensive nerve and axon regeneration among all groups, as evidenced by the significantly higher counts of S100 calcium‐binding protein B (S100‐β)‐positive cells (10,152 ± 986.00) and Neurofilament Protein 200 (NF200)‐positive cells (11,517 ± 795.70). Corroborating these histological findings, functional analysis demonstrated that the ADSC/SCNT/RAM group achieved the highest sciatic nerve function index (SFI) at 12 weeks post‐surgery (−58.06 ± 1.46), a value comparable to the Autograft group (−57.73 ± 1.80). This strategy proposes a promising tissue‐engineered alternative to autografts for nerve repair.
Immune checkpoint inhibitors (ICI) have demonstrated prolonged efficacy in certain melanoma patients, yet a significant portion of patients do not experience clinical improvement, with the mechanisms underlying this resistance still not fully understood. Using established cell markers, we partitioned the single-cell transcriptome into clusters, finding a notable link between NK cells and patient response to immunotherapy. We further identified four distinct subpopulations of NK cells, profiling marker gene sets and unique biological functions associated with each subpopulation. This analysis provides insights into the trajectory of NK cell development and differentiation, along with identifying the transcription factors driving these processes. The study pinpointed NK cluster 01 as pivotal in influencing patient sensitivity and prognosis during immunotherapy. Single-cell transcriptome and spatial transcriptomics (ST) analysis revealed the proximity of NK cluster 01 cells to melanoma cells, hinting at a potential regulation of cell-cell interaction via the IFN-II signaling pathway network. ST analysis revealed the spatial arrangement and interaction of NK cluster 01 cells with melanoma cells. This study explores the feasibility of targeting NK cluster 01 cells with small molecule drugs via molecular docking, offering a promising approach to bolster the clinical utility of NK cell therapy. We comprehensively analyze the heterogeneity of NK cells within melanoma, elucidate the potential regulatory interactions between NK cells and other microenvironmental components, and establish a basis for the future clinical utilization of distinct NK cell subsets as therapeutic targets.
