Accurate prediction of pathological complete response (pCR) following neoadjuvant therapy is crucial for personalized treatment planning in patients with locally advanced rectal cancer (LARC). This study aimed to explore the application of 3D transrectal ultrasound (TRUS) for this purpose. In this study, 538 LARC patients from five hospitals were enrolled and divided into training (n = 348), internal validation (n = 87), and external validation (n = 103) cohorts. Using pretreatment 3D TRUS data of the rectal tumors, a deep learning framework comprising an automated segmentation model and a pCR prediction model was constructed and validated. The segmentation model achieved an excellent Dice score of 0.89. The predictive model, when trained on data sampled at 9° or 18° intervals, yielded area under the curve (AUC) values of 0.92 (internal validation) and 0.85-0.86 (external validation), with accuracies of 86.2%-90.8% and 81.6%-83.5%, respectively; decision-curve analysis also demonstrated clinically meaningful net benefits. This study presents the first deep learning framework based on 3D TRUS for pCR prediction in LARC patients, offering an automated and reproducible tool that may support clinical decision-making in rectal cancer management. However, further validation in larger, multi-device cohorts is essential before clinical application.
In this study, we unexpectedly found that the immunoglobulin kappa light chain (Igκ) was expressed in normal cardiomyocytes of mice and humans, especially in intercalated discs (ICDs). Cardiomyocyte-specific knockout of Igκ in mice results in reduced myocardial contraction, atrioventricular block (AV block), and even sudden death. Histological analysis revealed that Igκ knockout in cardiomyocytes leads to structural disorder of ICDs, dissemination of the cytoskeleton proteins desmin and F-actin, as well as the loss of desmoplakin (DSP), N-cadherin, and connexin 43 (Cx43) on ICDs. Mechanistically, Igκ can bind to and stabilize plectin, a cytoskeleton-cross-linking protein that facilitates the assembly of desmin‒actin networks that maintain normal cytoskeletal architecture. Igκ can also anchor desmin to the DSP through plectin, thereby stabilizing the integrity of the ICDs. This molecular interplay critically reinforces cardiomyocyte cohesion and maintains structural and functional homeostasises. Our findings are the first to identify cardiomyocyte-expressed Igκ as a novel ICD-related molecule that participates in cardiomyocyte contraction and conduction by stabilizing plectins. Importantly, this work extends current arrhythmogenic cardiomyopathy (ACM) pathogenic models by revealing that ablation of the non-desmosomal gene Igκ disrupts ICD integrity, uncovering a new mechanism for non-desmosomal gene-related ACM and highlighting Igκ as a potential target for therapeutic investigations.
Mitochondrial dysfunction is one of the earliest pathological features of Alzheimer's disease (AD), preceding overt neurodegeneration and cognitive decline. Amyloid-β (Aβ) accumulation has long been considered a central pathogenic event in AD, yet how Aβ toxicity is mechanistically linked to mitochondrial impairment during early disease stages remains incompletely understood. To address this, we combined multi-omics with in vivo and in vitro genetic interventions. Here, we show that malic enzyme 3 (Me3) links Aβ aggregation to mitochondrial dysfunction in APP/PS1 mice and neuronal cells. At ultra-early and early AD stages (3 and 6 months), Me3 was upregulated and accumulated within mitochondria, where it colocalized with Aβ42 and physically interacted with it, an association linked to oxidative stress and impaired mitophagy. Knockdown of Me3 reduced mitochondrial reactive oxygen species, improved mitochondrial morphology, and alleviated mitophagy defects in both cellular and mouse models, with statistical significance across these functional measurements (p < 0.05). These results suggest that Me3 functions not only as a metabolic responder to Aβ-associated stress but also as a contributor to early mitochondrial pathology. By identifying the Aβ-Me3 axis, this study provides mechanistic insight into early mitochondrial dysfunction in AD, while further validation in human AD samples remains necessary.
Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 screening has become a central technology in functional genomics, enabling genome-scale interrogation via pooled perturbations. Early CRISPR screens employed survival or simple phenotypic readouts to identify essential genes and drug resistance mechanisms. However, as biological questions have shifted toward understanding regulatory networks, cellular heterogeneity, and context-dependent gene functions, there has been increasing demand for screening strategies capable of capturing complex cellular phenotypes beyond cell fitness. Recent advances in single-cell sequencing, high-content imaging, and spatial transcriptomics have expanded the resolution of CRISPR screening by enabling multidimensional phenotypic characterization following genetic perturbation. By integrating pooled perturbations with diverse readouts, these approaches systematically map targeted gene edits to transcriptional states, cellular phenotypes, and microenvironmental contexts. Meanwhile, innovations in library design, delivery, and computational pipelines have further improved the robustness and interpretability of high-content screening platforms. This review synthesizes the methodological evolution of CRISPR screening, emphasizing advances in perturbation strategies, delivery systems, and multimodal readouts. Representative applications spanning oncology, immunotherapy, developmental biology, neurobiology, and infectious diseases are delineated to demonstrate refined gene network annotations. Additionally, existing technical bottlenecks, such as scalability, cost constraints, and in vivo limitations, are critically assessed. Finally, future directions are proposed to facilitate the development of precise medicine.
Mesenchymal stromal cells (MSCs) are multipotent stromal cells with broad applications in regenerative medicine. The therapeutic effects of MSCs are increasingly attributed to immunomodulation, tissue repair and regeneration, paracrine effects, and mitochondrial transfer, rather than multilineage differentiation alone. Accumulating preclinical and clinical studies have demonstrated the therapeutic potential of MSCs across diverse disease settings, but clinical efficacy remains variable. However, differences in tissue source, donor age, expansion history, potency assays, and the disease microenvironment continue to limit reproducibility and clinical translation. Despite these advances, a comprehensive synthesis linking MSCs biology, mechanisms of action, disease-specific evidence, and clinical translation is still lacking. This review summarizes the biological characteristics and heterogeneity of MSCs, and discusses the specific evidence and therapeutic mechanisms of MSCs in treating diseases such as musculoskeletal, cardiovascular, immune-mediated, neurological, respiratory, and other diseases. We also summarize the major clinical transformation challenges of MSCs in treating various diseases. Additionally, we further explore emerging strategies to enhance the activity of MSCs, including pretreatment and activation, lifestyle intervention, and genetic engineering techniques. This review provides a comprehensive framework for developing safer, standardized, and clinically transformative therapies based on MSCs.
Full-thickness wounds remain a major clinical challenge, and tissue-engineered skin using epidermal stem cells (EpiSCs) offers a promising strategy for bio-inspired reconstruction. In this randomized controlled trial, 232 patients with full-thickness defects (≥ 9 cm2) received either standard treatment (acellular dermal matrix [ADM] + split-thickness skin graft [STSG], n = 112) or cell therapy (ADM + STSG + EpiSC suspension, n = 120). Autologous EpiSCs were prepared intraoperatively in a single step, avoiding prolonged in vitro expansion. The primary outcome was 6-month scar quality assessed by the Vancouver Scar Scale (VSS), while secondary outcomes included 2-week healing and 3-month recurrence. Compared with standard treatment, cell therapy produced significantly lower total VSS scores (median 3.0 vs. 6.0, p < 0.001) and improved complete wound healing at 2 weeks (80/120 [66.7%] vs. 61/112 [54.5%]; adjusted odds ratios [OR] = 1.87, 95% confidence interval [CI] 1.06-3.28, p = 0.029). Complete healing at 2 weeks was associated with reduced 3-month recurrence (adjusted OR = 0.26, 95% CI 0.09-0.75, p = 0.012). These findings support intraoperative EpiSC-enhanced composite grafting as an effective and clinically translatable approach for rapid closure and improved long-term scar outcomes.
Patent foramen ovale (PFO) is a highly prevalent cardiac abnormality, affecting approximately 25% of the population worldwide. Although a PFO may be discovered incidentally and remain asymptomatic throughout life, it can also be a conduit for paradoxical emboli or vasoactive substances and has been associated with neurological disorders, including cryptogenic stroke, transient ischemic attack, migraine, epilepsy, as well as systemic embolism, hypoxemic disorders, and decompression sickness, particularly in individuals with high-risk anatomy or traditional cardiovascular risk factors. However, accurate identification of pathogenic PFO remains a challenge in current clinical practice, and the indications, timing, and long-term safety of PFO closure remain controversial. In this review, we summarize the epidemiology, risk factors, pathophysiological mechanisms, clinical manifestations, diagnostic strategies, and management of PFO, with particular emphasis on risk stratification, emerging imaging tools, novel closure devices, procedure-related complications, and the application of closure in special populations. In addition, we focused on the current evidence regarding when and who may benefit from PFO intervention and the surveillance of postprocedural complications. Overall, this review provides a comprehensive overview of the current findings and future perspectives for the management of PFO.
Postoperative delirium (POD) is a common complication in older surgical patients and substantially worsens clinical outcomes, yet existing intraoperative electroencephalography (EEG) monitoring tools lack spatial and temporal specificity, creating a need for interpretable biomarkers. We prospectively analyzed 32-channel intraoperative EEG from 71 patients aged ≥ 60 undergoing noncardiac surgery, trained an interpretable spatiotemporal convolutional network (ST-CN), derived a best temporal filter (BTF), and evaluated model performance with region-specific tests and independent external validation. The ST-CN classified POD with 97.52% accuracy and an ROC of 0.996. The BTF alone discriminated POD with an AUC of 0.911 and achieved 85.12% accuracy using frontal EEG alone. It captured a distinct 2-12 Hz (δ-θ-α) oscillation in a spindle-like envelope, which occurred at a significantly higher rate in POD patients (4.62 ± 0.15 vs. 3.88 ± 0.13 waves/min in the frontal region) and differed in central frequency and spectral power. Independent external validation further confirmed robust generalizability, with frontal EEG achieving 89.01% accuracy and an AUC of 0.950. This framework enables accurate, interpretable POD risk stratification and identifies a reproducible frontal EEG biomarker, supporting objective intraoperative early warning and individualized perioperative care.
Ergothioneine (EGT) is a diet-derived sulfur-containing histidine derivative with antioxidant, anti-inflammatory, and cytoprotective properties. In humans, its absorption, tissue distribution, and long-term retention are mainly mediated by the organic cation transporter OCTN1, which may help explain its accumulation in metabolically active and stress-prone tissues. Increasing experimental and clinical evidence links EGT to neurodegenerative, cardiovascular, metabolic, inflammatory, and age-related disorders, although its therapeutic value still requires confirmation in larger and longer-term clinical studies. This review summarizes the physiological functions of EGT, its OCTN1-dependent transport and tissue distribution, and the molecular mechanisms by which it may modulate oxidative stress, inflammation, mitochondrial function, and cellular senescence. Particular attention is given to the dual role of the OCTN1/EGT axis in aging and tumor biology. In addition, we briefly discuss current biosynthetic and biomanufacturing strategies as practical tools for obtaining high-purity EGT for mechanistic, preclinical, and translational studies. By placing production strategies in the context of biomedical research needs, this review aims to provide a medically oriented overview of EGT biology and its potential translational applications.
Autoimmune diseases (ADs) arise from the breakdown of immune tolerance, yet their clinical course, organ involvement, and treatment responses vary markedly across systemic and organ-specific autoimmune disorders. This diversity cannot be explained by a single pathogenic mechanism or by conventional serological categories alone. Here, we synthesize evidence that autoimmunity is shaped by interacting molecular and cellular regulatory layers, including immunometabolic rewiring, epigenetic remodeling, posttranscriptional regulation, and functional reprogramming of immune and tissue-resident cells. These processes influence pathogenic T-cell and B-cell differentiation, myeloid activation, stromal and epithelial dysfunction, tissue-specific inflammation, and intercellular communication. We further discuss how multidimensional readouts, including metabolites, methylation and chromatin signatures, RNA-based markers, and reprogrammed immune-cell phenotypes, may support diagnosis, disease-activity assessment, prognosis, and therapeutic stratification. From a therapeutic perspective, this framework supports a shift from broad immunosuppression toward mechanism-matched interventions targeting metabolic vulnerabilities, epigenetic regulators, RNA-control pathways, and pathogenic cellular states, including emerging cell-based approaches. We propose that ADs can be viewed as dynamic network disorders in which molecular mechanisms, biomarkers, and therapeutic opportunities are closely connected. Integrating longitudinal multiomics, single-cell and spatial profiling, and standardized biomarker validation may help translate autoimmune heterogeneity into actionable precision-medicine strategies for more individualized care.
Neoadjuvant immunochemotherapy has transformed treatment paradigms in non-small cell lung cancer, yet its applicability in limited-disease small cell lung cancer (LD-SCLC) remains unknown. Here, we report findings from LungMate-006, the first prospective Phase II trial evaluating PD-1 blockade with tislelizumab combined with platinum-etoposide in LD-SCLC. Among 15 enrolled patients, the regimen achieved an objective response rate of 66.7% and enabled curative-intent surgery in 60% of patients, all with R0 resection. Major pathological response and pathological complete response occurred in 44.4% and 33.3% of surgical cases, respectively. Median event-free and overall survival reached 21.7 and 32.7 months. Metabolic response by PET-CT more accurately predicted pathological remission than radiographic shrinkage alone. Bulk RNA sequencing revealed a treatment-induced transition from a metabolically active baseline state toward an extracellular matrix-remodeled, CAF- and macrophage-enriched microenvironment, alongside upregulation of genes associated with tumor stemness and immune evasion. These transcriptomic changes suggest stromal-myeloid-mediated adaptive resistance in non-MPR tumors and highlight potential avenues for refining perioperative immunochemotherapy strategies in SCLC.
Cardiovascular diseases (CVD) remain a leading cause of death globally, yet assessing the combined impact of multiple metabolic disorders on CVD risk is challenging due to a lack of comprehensive tools. We aimed to develop and validate a novel CVD screening model based on metabolic clustering networks. Using data from 4066 adults enrolled in the STONE study (China) through stratified sampling, we constructed a holistic network map of metabolic health, incorporating obesity, dyslipidemia, glucose disorders, and hepatic, renal, thyroid, and bone conditions. Through network analysis, 12 central and clinically accessible indicators-including abdominal obesity, fatty liver, LDL-C, HbA1c, eGFR, TSH, and bone density-were selected to establish the CardioMet12 scoring system, with an AUC of 0.771 (95% CI: 0.733-0.808), sensitivity of 0.756, and specificity of 0.732. A score above 56 corresponded to an observed CVD prevalence exceeding 50% in the STONE cohort. External validation in an independent US population-based epidemiological sample demonstrated satisfactory performance. Higher scores were significantly associated with metabolic syndrome diagnosis and advanced cardiovascular-kidney-metabolic staging in both cohorts. In conclusion, CardioMet12 is a robust metric that captures the complex interplay among multiple metabolic disorders, offering a comprehensive and proactive tool for enhanced CVD screening and risk evaluation.
Exercise is a low-cost lifestyle intervention that can prevent and alleviate various diseases. It is a potent physiological stimulus that activates conserved molecular signaling pathways. Through the coordinated integration of multiple molecules, pathways, and systems, it leads to systemic health benefits. However, most studies focus on individual systems or molecular mechanisms, lacking systematic integration of the cross-system regulation induced by exercise. We summarize the molecular mechanisms of exercise in the musculoskeletal, cardiovascular, nervous systems, among others. Exercise induces the release of exerkines (e.g., irisin, interleukin-6, and brain-derived neurotrophic factor) and extracellular vesicles, which activate key signaling pathways to enhance mitochondrial function, metabolism and physiological adaptation, while suppressing inflammation and oxidative stress, thereby alleviating diseases and delaying aging through cross-system coordination. We further explore exercise-induced adaptive regulation in extreme environments, including microgravity, hyperbaria, and hypoxia, offering a multifaceted perspective on organismal health regulation. Finally, we outline the prospects and challenges of multiomics, artificial intelligence-driven precision medicine, personalized exercise prescriptions, and exercise mimetics. Overall, this review provides a more integrated perspective on the molecular basis of exercise and offers directions for future mechanistic and translational studies.
The heart and brain maintain systemic homeostasis through continuous bidirectional communication mediated by neural regulation, biochemical signaling, and mechanical transduction. Disruption of this communication is increasingly recognized as a contributor to cardiovascular, neurological, cognitive, and psychological disorders. Growing evidence indicates that autonomic imbalance, neuroimmune activation, hemodynamic disturbances, inflammation, altered interoception, and brain network remodeling can mediate cross-organ injury between the heart and brain. These mechanisms underlie a spectrum of heart-brain comorbidities, but their shared pathways, disease-specific manifestations, and integrated management strategies remain insufficiently understood. In this review, we summarize the physiological basis of heart-brain interactions, focusing on neural, biochemical, and mechanical pathways. We then discuss six representative disorders: Takotsubo syndrome, epileptic heart, stroke-heart syndrome, heart failure, cardiac dementia, and psychocardiology. For each condition, we highlight key epidemiological features, pathophysiological mechanisms, and integrated heart-brain management strategies. We also review clinical trials targeting heart-brain interactions, including autonomic modulation, biomarker monitoring, neuroimaging assessment, exercise therapy, psychological intervention, and pharmacological therapy. By integrating basic mechanisms, clinical phenotypes, and translational advances, this review provides a systematic framework for understanding heart-brain comorbidities and advancing precision heart-brain medicine.
Solitary distant metastasis (SDM) is a distinct oligometastatic subtype associated with a relatively favorable prognosis. This multicenter retrospective study including 139 non-small cell lung cancer (NSCLC) patients with SDM (2019-2024) was conducted to investigate prognostic factors for multiple metastases and identify the critical time window for local treatment. Statistical analyses included Cox regression, Kaplan-Meier survival analysis, and restricted mean survival time (RMST). Results showed that driver gene status, tumor staging, systemic therapy, and local treatment were significantly associated with progression-free survival (PFS). Notably, only early local treatment significantly delayed the onset of multiple metastases (hazard ratio [HR] = 0.55, 95% confidence interval [CI]: 0.30-0.99, p = 0.05), prolonging the median time to multiple metastases by nearly 14 months (36.4 vs. 22.6 months). Kaplan-Meier and RMST analyses revealed no distinct plateau phase, indicating the absence of an absolute temporal cutoff between "true oligo-metastasis" and "transitional metastasis," which suggests that SDM in advanced NSCLC is an inherently indolent disease state. Using the maximum selected rank statistics method, an optimal cutoff of 138 days was identified to distinguish early from delayed local treatment, which requires further prospective validation.
As a critical hub in the thalamo-cortical circuit, the human thalamus engages in a spectrum of fundamental and advanced brain functions through widely distributed circuits. Clinically, dysfunction of thalamo-cortical circuits is shown to be profoundly implicated in a wide range of neurological and psychiatric diseases. Nevertheless, the neuroanatomical substrates governing these functions of the thalamus have rarely been directly mapped in humans. Here, we overviewed the acute responses of direct electrical stimulation (DES) delivered to the distributed thalamus sites in 52 epilepsy patients admitted for presurgical stereoelectroencephalography. Specifically, DES of the thalamus evoked a broad spectrum of in-situ responses spanning fundamental functions such as sensory and motor processing, extending to complex neural operations encompassing neurovegetative regulation, cognitive processing, emotional modulation, and multimodal responsiveness. Moreover, through the integration of DES with functional human connectome (n = 1000), we found an intra-thalamic intrinsic functional network associated with each specific clinical response. Our data provide direct substantiation for the complex functional architecture of the human thalamus, advancing current understanding of its role and potentially culminating in targeted therapeutic strategies tailored to ameliorate symptom-specific neural circuit disorders.
Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening disease characterized by hyperinflammation. Primary HLH (primary HLH), resulting from genetic mutations, is a subtype of HLH. The mutation spectrum of primary HLH-associated genes has not been well determined in China. This study aimed to explore the mutation spectrum of 12 primary HLH-associated genes in a multicenter database. Medical records and gene sequencing data of 1224 HLH patients were retrieved from January 2014 to September 2024, and gene mutations were analyzed. Genetic variants were observed in 350 (28.59%) patients. Among them, 108 patients had a definitive genetic diagnosis, 227 patients carried single heterozygous variants, and 15 patients carried digenic/polygenic heterozygous variants. A total of 275 different variants were identified, and missense variants were most common. Variants in UNC13D were most common, followed by LYST and PRF1, while variants in MAGT1, ITK, and CD27 were rare. For UNC13D, LYST, and PRF1, variants c.2588G>A (p.Gly863Asp), c.368A>G (p.His123Arg), and c.1349C>T (p.Thr450Met) were most common, respectively. We described the mutation spectrum of primary HLH-associated genes in a largest Chinese cohort and found some mutation specificity for Chinese HLH patients. Our data might help design sequencing panels, interpret sequencing results, and understand genetic background of Chinese HLH patients.
Locally advanced head and neck carcinoma remains associated with high morbidity and long-term survival below 50%. Treatment commonly relies on cisplatin-based chemoradiotherapy, which is effective but frequently associated with significant acute and chronic systemic toxicities. Therefore, safer and more effective therapeutic strategies are urgently needed. FLASH radiotherapy (RT) has emerged as a promising irradiation modality because of its potential to reduce damage to healthy tissues while preserving antitumor efficacy. Here, we investigated the anticancer activity of the Topoisomerase (Topo)-II inhibitor ARN-24139, alone and combined with FLASHRT, in human papillomavirus-negative SCC-25 head and neck carcinoma biomodels. Antitumor activity was assessed in 2D cell cultures using viability, apoptosis, clonogenic, wound-healing, and γH2AX assays, as well as in SCC-25 3D spheroids and in chorioallantoic membrane (CAM) tumor models. ARN-24139 induced dose-dependent cytotoxicity in SCC-25 cells, with IC50 values of 7.3 ± 0.8 µM at 48 h and 7.2 ± 0.5 µM at 72 h, while showing limited toxicity in healthy HBEpC cells. Sequential low-dose FLASH-RT followed by ARN-24139 enhanced antitumor activity, reducing cell viability at 4 Gy after 8 days and decreasing tumor growth and Ki67 expression in CAM models. These proof-of-concept findings support further investigation in more clinically representative and mechanistically informative HNSCC models.
