To clarify the therapeutic targets and signaling pathways of Actinidia arguta total flavone (AATF) in gout treatment, we integrated network pharmacology and in vivo experiments. Network pharmacology was applied to screen AATF's anti-gout targets, construct protein-protein interaction (PPI) and drug-component-disease-target-pathway networks, and conduct gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. A dual gout rat model was established using potassium oxonate, adenine, and monosodium urate (MSU) crystals, followed by assessments including toe swelling measurement, hematoxylin-eosin (HE) staining of kidney and joint tissues, digital radiography (DR) imaging, detection of serum uric acid and xanthine oxidase (XOD) activity, enzyme-linked immunosorbent assay (ELISA) of inflammatory factors, and Western blot validation of key targets. Network pharmacology revealed that AATF modulates inflammatory responses and the tumor necrosis factor (TNF) pathway via core targets including albumin (ALB), TNF, interleukin-6 (IL-6), and tumor protein 53 (TP53). In vivo experiments showed that AATF significantly ameliorated renal and joint pathological damage, reduced serum uric acid/XOD activity, downregulated serum interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-6, cyclooxygenase-2 (COX-2) levels, and inhibited joint nuclear factor kappa B (NF-κB), extracellular signal-regulated kinase 1 (ERK1), matrix metalloproteinase 9 (MMP9) expression. Collectively, AATF exerts anti-gout effects through multi-target and multi-pathway mechanisms linked to the TNF signaling pathway, providing critical preliminary evidence for its preclinical development.

IDH mutant gliomas, driven by the oncometabolite 2-hydroxyglutarate (2-HG), are associated with profound neurological morbidity and premature mortality. To address the unmet therapeutic needs, we investigated the mechanistic interplay between MTHFD2-driven one-carbon metabolism and ferroptosis susceptibility in these tumors. Our findings revealed that MTHFD2 upregulation, mediated through loss of m5C modification in chromatin associated RNAs (caRNAs)—establishes a metabolic vulnerability to ferroptosis. Crucially, we identified TAF15 as a pivotal RNA-binding protein that orchestrates the spatial recruitment of TET2 by bridging NSUN5-mediated RNA m5C methylation. Therapeutically, combinatorial targeting of MTHFD2 with its selective inhibitor and the hypomethylating agent decitabine induced ferroptosis in patient-derived IDH-mut glioma organoids, demonstrating potent ferroptosis activation. This work delineates an RNA epitranscriptomic-metabolism axis in glioma pathogenesis and provides a translational roadmap for exploiting metabolic dependencies in IDH-driven malignancies. Not applicable.

Background:Systemic lupus erythematosus (SLE) is characterized by immune dysregulation driven in part by aberrant CD4+ T cell activation and defective autophagy. Although exosomes are increasingly recognized as mediators of immune communication, the contribution of exosomal long non-coding RNAs (lncRNAs) to T cell dysfunction in SLE remains poorly defined. This study investigated the pathogenic role and mechanism of plasma exosomal lncRNA XIST in regulating CD4+ T cell autophagy and activation. Methods:Quantitative reverse transcription PCR (qRT-PCR) was used to quantify lncRNA XIST levels in plasma exosomes, while Western blotting, flow cytometry, and enzyme-linked immunosorbent assay (ELISA) were employed to assess the effects of exosomes on CD4+ T cell function. Gain- and loss-of-function approaches were applied to elucidate the underlying molecular mechanisms. Results:LncRNA XIST was significantly overexpressed in SLE plasma exosomes and positively correlated with disease activity. These exosomes enhanced lncRNA XIST expression in CD4+ T cells from healthy controls (HCs), leading to suppressed autophagy and increased activation. Similar effects were observed with engineered HC plasma exosomes overexpressing lncRNA XIST. Conversely, silencing lncRNA XIST in SLE CD4+ T cells promoted autophagy and reduced activation, although these changes were reversed upon treatment with autologous plasma exosomes. Mechanistically, exosomal XIST functioned as a molecular sponge for miR-98, upregulating RICTOR and activating the Akt/mTOR signaling pathway, thereby modulating CD4+ T cell function. Conclusion:Plasma exosomal lncRNA XIST impairs CD4+ T cell autophagy and promotes activation through the miR-98/RICTOR/Akt-mTOR axis, representing a mechanistically defined candidate biomarker and potential therapeutic target in SLE.

Neutrophil extracellular traps (NETs) are increasingly recognized for their critical roles in tumor progression and tumor immune microenvironment (TIME) modulation, yet their functions in glioma remain incompletely understood. Leveraging 68 NET-associated regulators, we identified two distinct NET-based molecular subtypes. The G1 subtype exhibited an elevated NET expression profile, immunosuppressive TIME, and was associated with unfavorable patient outcomes. MMP9, TLR8, and LILRB2 were pinpointed as key regulators and a prognostic model was constructed with robust predictive performance. These regulators could promote NET formation, suppress T-cell activity and facilitate immune evasion in glioma. Mechanistically, the NEAT1/miR-149-5p/MMP9, TLR8 and LILRB2 axis demonstrated to promote glioma progression. Patients with high expression levels of these regulators were predicted to correlate with better chemotherapy response but poorer immunotherapy outcomes. In summary, our results identify MMP9, TLR8, and LILRB2 as pivotal regulators of glioma malignancy and TIME remodeling, with significant implications for prognosis and the development of targeted therapies.

Neutrophil extracellular traps (NETs) are increasingly recognized as key regulators of tumor progression, yet the molecular circuitry that governs their induction in cancer remains elusive. Here, we identify the RNA-binding protein RBFOX2 as a tumor suppressor that curtails glioma growth by coordinately restraining tumor cell proliferation and NETosis. RBFOX2 expression is markedly reduced in glioma and positively correlates with patient survival. Mechanistically, RBFOX2 binds to 5-hydroxymethylcytidine (5hmC)-modified sites within PDGFB mRNA and promotes its decay, thereby dampening AKT-SP1 signaling and repressing CSF3 transcription. This repression limits neutrophil-mediated NET formation in the tumor microenvironment, as confirmed in PAD4-/- mice and upon CSF3 neutralization. Collectively, our study uncovers a 5hmC-dependent post-transcriptional mechanism linking RBFOX2 to NETosis control and glioma suppression, revealing RBFOX2 as a potential biomarker and therapeutic lever and establishing a broader paradigm in which RNA-binding proteins couple post-transcriptional RNA modification and immune regulation in tumor evolution.

Metabolic signals critically shape innate immune responses. Through pharmacological screening of metabolic pathways, we identified aspartate metabolism as a key regulator of cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) signaling. Genetically or aminooxyacetic acid-mediated (AOA-mediated) pharmacologically reducing aspartate levels markedly potentiated the cGAS-STING pathway, leading to stronger upregulation of type I interferons and interferon-stimulated genes. Mechanistically, disruption of de novo pyrimidine synthesis, a major downstream pathway of aspartate, induced mtDNA replication stress and increased mtDNA double-strand breaks, promoting mtDNA release into the cytosol. Cytosolic mtDNA synergized with cGAS-STING agonists to upregulate Z-DNA binding protein 1 (ZBP1), which recruits RIPK1/3 to sustain IRF3 phosphorylation, forming a positive feedback loop that amplifies innate immune signaling. In immunocompetent mouse models, AOA enhanced the antitumor efficacy of STING agonists, chemotherapy, or radiotherapy, whereas aspartate supplementation abrogated these effects. Consistently, aspartate levels negatively correlated with antitumor immunity in colorectal cancer patient samples. Together, our study identifies aspartate-pyrimidine metabolism as a critical metabolic checkpoint that licenses STING signaling by enabling mtDNA stress to cooperate with agonist stimulation, driving type I interferon-dependent ZBP1 induction and feed-forward amplification of STING signaling, thus offering a promising strategy to enhance antitumor immunity.

IntroductionGlioblastoma (GBM) is a highly lethal malignancy driven by glioma-initiating cells (GICs). While GICs are known to profoundly remodel tumor microenvironment (TME) to promote progression and immune evasion within the vascular niche, the specific transcriptomic reprogramming and alternative splicing events driving their evolution from neural stem cells (NSCs), and how these intrinsic cellular state changes dictate multi-cellular immunosuppressive networks and checkpoints, remain poorly understood. Unraveling these complex tumor-vascular-immune interactions is critical for identifying novel vulnerabilities and developing effective immunotherapies.MethodsTo decode the GICs’ evolutionary trajectory, we integrated RNA-seq and alternative splicing analysis of NSCs and patient-derived GIC cohorts. The malignant progression was mapped using scRNA-seq pseudotime analysis, and key targets were validated across clinical TCGA cohorts. Furthermore, we employed the large-scale single-cell foundation model, Geneformer, to perform in silico genetic perturbations, integrating it with interactome inference to decipher TME communication. Finally, the proposed tumor-endothelial-T cell multi-cellular axis was functionally validated utilizing in vitro tumor-HUVEC co-culture systems, qPCR, and FACS-based T cell activation (NFAT-Jurkat) assays.ResultsOur multi-omics re-analysis identified extensive alternative splicing and transcriptional reprogramming during GICs evolution, pinpointing SLC1A3 as a core gene significantly upregulated along the malignant pseudotime trajectory and strongly correlated with poor clinical prognosis in GBM. AI-driven in silico virtual knockout utilizing Geneformer revealed that SLC1A3 acts as a master regulator of tumor network stability. Interactome analysis demonstrated that SLC1A3hi tumor cells exhibit intensive communication with endothelial cells via specific ligand-receptor axes (e.g., TNC-ITGB1, PTN-SDC3). In vitro assays confirmed that endothelial cells were educated by SLC1A3hi tumor cells that undergo malignant transition, drastically upregulating immune-suppressive factors, including CD274, TGFB1, IL10, and IDO1. Crucially, tumor-specific knockdown of SLC1A3 dismantled this vascular-immune suppressive niche, significantly restoring T cell activation in a multicellular co-culture model.DiscussionOur findings establish SLC1A3 not merely as an intrinsic driver of glioma development, but as a critical upstream node orchestrating a cascading tumor-endothelial-T cell immunosuppressive axis. By leveraging AI-based foundation models alongside robust biological validation, we uncovered a novel mechanism of vascular-mediated immune evasion, highlighting SLC1A3 as a highly promising therapeutic target to reprogram the glioblastoma microenvironment and restore anti-tumor immunity.

BackgroundSchizophrenia (SCZ) pharmacotherapy relies on Western medications with limited efficacy/significant adverse effects. Baicalin (BA), a purified botanical monomer, shows promise as a safer multitarget antipsychotic candidate.ObjectiveTo study the effect and mechanism of Baicalin on MK-801 induced schizophrenia model mice.MethodsBehavioral assessments (water maze, open field, dark avoidance, forced swimming tests) evaluated emotional/cognitive functions in MK-801 induced schizophrenia mice. Histological staining analyzed hippocampal, prefrontal cortical, and striatal morphology. Serum inflammatory markers (NF-κB, IL-6, IL-1β, TNF-α) and oxidative stress indicators (SOD, MDA) were quantified by ELISA, alongside hippocampal neurotransmitter levels (DA, 5-HT, GABA, AChE). This study employed a network pharmacology approach to screen the mechanisms of action of baicalin in the treatment of schizophrenia. Western blotting determined hippocampal PI3K/Akt/GSK3β pathway protein expression.ResultsNetwork pharmacology analysis revealed that baicalin may exert its therapeutic effects in the treatment of schizophrenia through modulation of the PI3K-Akt signaling pathway. Versus model group, BA doses significantly decreased: IL-1β, IL-6, GABA, AChE, MDA, TNF-α, NF-κB; open field total distance; forced swimming immobility; dark avoidance errors (p < 0.05). Increased: DA, 5-HT; water maze platform crossings; dark avoidance latency (p < 0.05). Staining confirmed BA reduced cerebral oxidative stress/neuroinflammation. Western blot showed dose-dependent elevation of p-Akt/Akt and p-GSK3β/GSK3β ratios.ConclusionBaicalin may improve cognitive impairments in MK801-induced schizophrenia model mice through the PI3K/Akt/GSK3β signaling pathway, exhibiting anti-neuroinflammatory and neuroprotective effects.

Acinetobacter baumannii (A. baumannii), a very common pathogen, poses a significant public health threat due to its antibiotic resistance and long survival in healthcare environments. Both A. baumannii and carbapenem-resistant A. baumannii (CRAB) can spread through the air, increasing infection risks. Therefore, monitoring their presence in the air is of great significance, especially in hospitals. Herein, we developed a Chelex-100-LAMP-CRISPR/Cas12a (CLC) platform including DNA release and nucleic acid test. Combined with a wet cyclone sampler, the platform can detect airborne A. baumannii and its most common carbapenem-resistant gene, blaOXA-23, within 70 min. This CLC platform has also been proven to have a detection limit of 6 × 102 CFU of CRAB per test through simulated air samples. Moreover, this platform was also used to test five actual air samples from a tertiary hospital, and the results achieved perfect concordance with sequencing data, validating the platform’s accuracy and reliability. Therefore, the CLC platform showed great potential for the rapid, on-site detection of airborne A. baumannii and its carbapenem-resistant gene blaOXA-23, offering a valuable tool for infection control in healthcare environments.

RNA sequencing (RNA-seq) is a widely used and powerful technique for studying gene expression. Among the various protocols, SHERRY (sequencing hetero RNA-DNA-hybrid) profiles polyadenylated RNAs by direct tagging of RNA/DNA hybrids and offers a robust and economical way for gene expression quantification. Here, we present a detailed protocol for standard SHERRY library preparation from 200 ng of total RNA. We describe steps of RNA purification, reverse transcription, hybrid tagmentation, and library generation. We then detail procedures for sequencing and data analysis. For complete details on the use and execution of this protocol, please refer to Di et al.1.

Hearing loss is one of the most prevalent sensory disorders, but no commercial biological treatments are currently available. Here, we identify an East Asia-specific founder mutation, the homozygous c.220 C > T mutation in MPZL2, that contributes to a significant proportion of hereditary deafness cases in our cohort study. We find that the disease-causing mutation can be targetable by adenine base editors (ABEs) that enable A·T-to-G·C base corrections without DNA double-strand breaks. To demonstrate this, we develop a humanized mouse model (hMPZL2Q74X/Q74X) that recapitulates human MPZL2 deafness and leads to progressive hearing loss. A PAM-flexible ABE variant with reduced bystander and off-target effects (ABE8eWQ-SpRY:sgRNA3) is packaged in dual adeno-associated viruses (AAVs) and injected into the inner ear of hMPZL2Q74X/Q74X mice and effectively corrects the mutation. This treatment significantly restores hearing function, improves inner ear structural integrity, and reverses altered gene expression. Base editing may hold therapeutic potential for hereditary deafness, including most cases of MPZL2 deafness.

Dual blockade of the PD-1/PD-L1 axis, enabling tumor immune evasion, and the VEGF pathway, driving immunosuppression, represents a promising cancer immunotherapy strategy. Combining immune checkpoint inhibitors (ICIs) with antiangiogenics faces toxicity and cost limitations. Bispecific antibodies (BsAbs) targeting both pathways offer a solution. Preclinical and clinical studies demonstrate that simultaneous inhibition enhances antitumor immunity by reversing T-cell exhaustion, normalizing vasculature, and countering immunosuppression. Ivonescimab, a first-in-class PD-1/VEGF BsAb, exemplifies this approach. Approved in China (NMPA, May 2024) for EGFR-mutant non-squamous NSCLC post-TKI failure and included in national insurance (November 2024), it is under global evaluation in solid tumors. PD-1(L1)/VEGF BsAbs like ivonescimab represent a novel therapeutic strategy with potential for improved efficacy and mitigated toxicity compared to combination therapies. Ongoing trials will define broader applications.

To investigate the clinical significance of results in electrodiagnostic evaluations for carpal tunnel syndrome(CTS) and analyse the correlation between electrodiagnostic evaluations and physical examinations, 236 CTS patients and 96 healthy volunteers were included to accept electrodiagnostic evaluations. There were significant differences between CTS and control groups, mild and moderate patients, moderate and severe patients in CTS groups respectively, in terms of compound muscle action potential(CMAP), sensory conduction velocity of middle finger(SCV), distal motor latency(DML) difference between median nerve and ulnar nerve on the ring finger(△DML) (p<0.05).△DML has larger area under curve of ROC curve (0.974) than CMAP, SCV and DML(0.748, 0.950, 0.958), then △DML could be included as valuable test for diagnosing and classification of carpal tunnel syndromes. There was low correlation between electrodiagnostic evaluations and physical examinations (rho <0.4),so electrodiagnostic evaluation and physical examination should be combined to diagnose CTS.

Mastitis is a significant concern in both human and animal medicine. The causative agent S. aureus is one of the most challenging pathogens responsible for mastitis, and the rise of its antibiotic resistance underscores the need for alternative therapies. The SESN2/Nrf2 pathway, owing to its pivotal role in regulating cellular antioxidant defenses, which are critically disrupted during ferroptosis, has recently received less attention. However, whether C3G targets the SESN2/Nrf2 pathway remains unclear, which provides a dual mechanism for treating S. aureus-induced mastitis by reducing inflammation and safeguarding mammary epithelial cells (MECs) from ferroptosis. Using a mouse mastitis and MECs model, we investigated the therapeutic potential of C3G in alleviating S. aureus-induced mastitis, focusing specifically on its role in inhibiting inflammation and modulating ferroptosis through the SESN2/Nrf2 pathway. The results demonstrated the potential antimicrobial effects of C3G against S. aureus and MRSA, suppressed inflammatory responses by downregulating pro-inflammatory markers (IL-1β, IL-6, and TNF-α), and inhibited STAT2/STAT3 signaling. Furthermore, C3G modulates ferroptosis by activating the SESN2/Nrf2 pathway, reducing oxidative stress, and protecting mammary epithelial cells from ferroptosis-induced damage. This comprehensive approach highlights C3G's potential as a novel therapeutic strategy for managing mastitis, offering an effective alternative to antibiotics in addressing both bacterial infection and inflammation.

Acute pulmonary embolism (APE) is the third most frequent life-threatening cardiovascular disease. Which clinical parameters correlate with APE severity are still unclear. This study aimed to investigate the association between clinical parameters and APE severity, and to detect the parameters that could predict severe (intermediate high- and high-risk) APE. Clinical data from 612 hospitalized APE patients from two centres in China were collected. Univariate or multivariate ordinal logistic regression analyses were used to detect the relationship between clinical parameters and APE severity. Receiver-operating characteristic (ROC) curves were used to calculate the predictive values of single or combined parameters in predicting severe APE or 30-day mortality in a discovery cohort (n = 368) and a validation cohort (n = 244). Age (odds ratio [OR], 1.462; 95

Traditional contralateral seventh cervical nerve root (C7) transfer sacrifices the recovery of ulnar nerve. This cadaveric study, done in 20 upper extremities from 10 cadavers, looks at the feasibility of using a modified contralateral C7 transfer using lateral antebrachial cutaneous nerve to allow recovery of ulnar nerve. The deep branch of ulnar nerve was preserved with ipsilateral lateral antebrachial cutaneous nerve, while the distal end of ulnar nerve with its dorsal and superficial branches was transferred to contralateral C7 nerve root and the proximal end of ulnar nerve was anastomosed to both median and musculocutaneous nerves. The ipsilateral lateral antebrachial cutaneous nerve was separated and coapted with motor branch of ulnar nerve. The distance from the midpoint of connection of medial and lateral epicondyles of humerus (interepicondylar line) to the starting point of deep branch of ulnar nerve was significantly shorter than that to the branching point of the lateral antebrachial cutaneous nerve (p < 0.05). The differences in diameters between deep branch of ulnar nerve and main trunk and branches of lateral antebrachial cutaneous nerve were not significant (p > 0.05). The ratio of ulnar nerve axon count to combined total axon count of musculocutaneous nerve, main trunk of lateral antebrachial cutaneous nerve and branches to deep branch of ulnar nerve were above 1:3. In conclusion, the deep branch of ulnar nerve could be restored with lateral antebrachial cutaneous nerve, while contralateral C7 was transferred to median and musculocutaneous nerves through grafts of dorsal and superficial branches of ulnar nerve.

Glioma represents a predominant intracranial malignancy, with fewer than 40% of patients surviving beyond five years. MiRNA-21 antisense oligonucleotides (anti-miR-21) hold considerable promise for managing aggressive intracranial gliomas. Yet, challenges such as in vivo degradation, suboptimal brain targeting, and inadequate tumor penetration compromise the therapeutic efficacy of unmodified miRNA-21 inhibitors. To overcome these obstacles, we designed an orally-administered, brain-targeted anti-miRNA-21 lipid polymer micelle system (BTMLPMS), dually modified with Ang-2 and TAT. This modification ensures the stable delivery of anti-miR-21, leveraging the protective capacities of lipid polymer micelles to effectively impede glioma growth in vivo. In vitro studies confirmed that these micelles were efficiently uptaken by glioma cells, resulting in induced apoptosis. Importantly, in an orthotopic glioma xenograft model using nude mice, oral administration of these micelles activated pro-apoptotic proteins p53 and Caspase-3, effectively inducing apoptosis in tumor tissues and curbing tumor progression. Hematoxylin-eosin staining further validated the biocompatibility of the micelles. Collectively, our findings underscore the potential of the anti-miR-21 lipid polymer micelle system as a significant advancement in glioma gene therapy, paving the way for groundbreaking transformations in miRNA clinical applications.