Glioblastoma is an immunologically cold, treatment-refractory brain tumor in which a profoundly immunosuppressive microenvironment drives resistance to immunotherapy. Retrospective clinical series reporting unexpectedly prolonged survival after postoperative intracranial infections raise the possibility that a brief, spatially confined inflammatory burst within the central nervous system can mitigate this resistance. Tumor necrosis factor alpha (TNFα) and interferon gamma (IFNγ) are central inflammatory cytokines, yet how to harness such cytokine-rich inflammatory signaling in glioblastoma remains unclear. Here, integrating multi-omics, high-dimensional immune profiling, and orthotopic models, we show that TNFα plus IFNγ triggers immunogenic PANoptosis and reprograms the tumor microenvironment. Co-stimulation induces a fused pyroptosis-apoptosis-necroptosis program, increases calreticulin exposure and extracellular release of adenosine triphosphate and high-mobility group box 1, and shifts tumor-associated microglia and macrophages toward a pro-inflammatory, antigen-presenting state with enhanced effector and memory T-cell infiltration and activation. Mechanistically, IFNγ-STAT1-IRF1 signaling primes Z-DNA binding protein 1 (ZBP1), whereas TNFα induces SLC39A1-dependent zinc accumulation, oxidative stress, and Z-form nucleic acids that engage ZBP1 to assemble the PANoptosome and execute PANoptosis. In vivo, local intratumoral TNFα+IFNγ delivery suppresses tumor growth, prolongs survival, and restores responsiveness to PD-1 blockade. A liposomal formulation co-encapsulating zinc and IFNγ recapitulates this circuitry while avoiding systemic TNFα exposure and further improves checkpoint efficacy. These findings position ZBP1-centered PANoptosis as a druggable signaling node linking inflammatory and metal-ion cues to immune reprogramming and immunotherapy sensitization in Glioblastoma.

Glioblastoma (GBM) is the highly lethal intracranial tumor characterized by low survival rates and high recurrence, partly attributable to the challenges posed by the blood-brain barrier (BBB). To enhance therapeutic efficacy, the Exo-U2-Dox complex was engineered by functionalizing mesenchymal stem cell (MSC)-derived exosomes with the GBM-targeting aptamer U2 and integrating them with doxorubicin (DOX). This complex is designed to augment the sensitivity of GBM to chemo-radiotherapy. Here, it is found that Exo-U2 effectively accumulates in GBM-bearing mice, thereby inhibiting tumor progression. When administered in conjunction with DOX and radiation, Exo-U2-Dox increases DNA damage in GBM cells, and diminishes invasiveness. Mechanistically, Exo-U2 targets and inhibits the autophosphorylation of Epidermal growth factor receptor variant Ⅲ (EGFRvⅢ) in GBM cells, thereby activating the NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome-mediated pyroptosis pathway, which leads to increased expression of Gasdermin D (GSDMD) and Cysteine-aspartic acid protease-1 (caspase-1), ultimately suppressing GBM cell proliferation, migration, and invasion. Furthermore, the combination of Exo-U2 with X-ray treatment inhibits the expression of p53-binding protein 1 (53BP1), reduces phosphorylation of the Ataxia-Telangiectasia Mutated/Checkpoint kinase 2 (ATM/Chk2) pathway, resulting in the accumulation of DNA damage. Collectively, these findings underscore the potential of aptamer-functionalized exosomes in conjunction with DOX as a promising strategy for GBM treatment. This approach not only broadens the therapeutic applications of DOX but also provides a novel direction for targeted GBM therapies.

Glioblastoma (GBM) remains incurable due to the blood-brain barrier (BBB) limiting drug delivery and intrinsic/acquired resistance to temozolomide (TMZ), the first-line chemotherapy. Here, we developed Angiopep-2 (Ang)-modified natural killer cell-derived extracellular vesicles (Ang-NK-EV) for targeted TMZ delivery (Ang-NK-EV@TMZ) to address these bottlenecks. NK-EV were prepared via freeze-thaw extrusion of NK-92 cells, loaded with TMZ, and surface-functionalized with Ang to target LRP1 (highly expressed at the BBB and on GBM cells). Characterization confirmed Ang-NK-EV@TMZ exhibited spherical morphology, preserved EV markers (TSG101, CD9/63/81), and retained NK cell-derived immune factors (IFN-γ, GZMB). In vitro, Ang modification enhanced GBM cell uptake (2.5–3.2-fold vs. NK-EV) and BBB transcytosis (2.8–3.5-fold vs. free TMZ). Ang-NK-EV@TMZ reversed TMZ resistance by modulating STING/ mTOR/ MGMT signaling (via IFN-γ) and inducing apoptosis (elevated cleaved caspase-3, γ-H2AX). It also triggered immunogenic cell death (increased ATP, HMGB1) and polarized macrophages to M1-like phenotypes. In orthotropic GBM models, Ang-NK-EV@TMZ accumulated in brain tumors, inhibited growth (7.2-fold lower bioluminescence vs. PBS), and extended median survival (42 days vs. 18 days for PBS). No significant organ toxicity or hemolysis was observed. This platform integrates targeted chemotherapy and immune modulation, highlighting NK-EV’ potential for GBM therapy.

Introduction Gestational diabetes mellitus (GDM) occurs in pregnant women of advanced maternal age and is characterized by dysregulated elevation of blood glucose levels. Ginkgolide B (GB) is extracted from Ginkgo biloba leaves and possesses anti-inflammatory as well as antioxidant effects. This study aimed to assess the function of GB in gestational diabetes mellitus (GDM) and explore the underlying mechanisms. Methods Pregnant C57BL/KsJ-db/+ mice were assigned to the untreated GDM group or treated orally with GB at 100 or 200 mg/kg. Glucose tolerance, insulin sensitivity, fasting blood glucose, plasma insulin levels, and pregnancy outcomes were evaluated. Placental inflammation, oxidative stress, ferroptosis-related proteins, NLRP3 inflammasome components, and proteins involved in the Nrf2/HO-1 signaling pathway were assessed using ELISA, histological staining, immunofluorescence, biochemical assays, and Western blotting. Results Compared with the Control mice, GDM mice exhibited impaired glucose tolerance and insulin sensitivity, elevated fasting blood glucose and plasma insulin levels, increased placental inflammation and oxidative stress, enhanced ferroptosis, and reduced litter size. GB treatment significantly improved these metabolic and pregnancy-related abnormalities, attenuated inflammatory cytokine production and placental injury, reduced reactive oxygen species accumulation and lipid peroxidation, and restored GPX4 and SLC7A11 expressions. In addition, GB inhibited NLRP3 inflammasome activation, while restoring the expression of Nrf2, HO-1, and NQO1 in placental tissues. Discussion This study showed the beneficial effects of GB in GDM, suggesting potential therapeutic approaches for preventing the progression of GDM management.

Medulloblastoma, the most common malignant pediatric posterior fossa tumor, exhibits metabolic reprogramming and tumor immune microenvironment heterogeneity in non-WNT/non-SHH subgroups, yet the interplay between these features remains poorly defined. Revealing potential molecular features and hidden therapeutic target, we integrated the transcriptomic data of non-WNT/non-SHH medulloblastoma from GEO and EMBL-EBI databases. Through examination of the Grp3/4 transcriptional continuum, we identified PCK2 (a key gene in the TCA cycle with the highest correlation with continuum score). We validated PCK2 as a driver of proliferation, migration, invasion, glycolysis, and M2 macrophage polarization in MYC-amplified MB cells through short hairpin RNA knockdown experiments. Together, our findings establish metabolic-TIME crosstalk as a prognostic determinant and proffer PCK2 as a therapeutic target for aggressive MB subtypes, offering insights into metabolic subtyping and precision therapy strategies to improve clinical outcomes.

This study aimed to develop a predictive model for assessing the risk of hemorrhagic transformation (HT) following mechanical thrombectomy (MT) in patients with acute ischemic stroke (AIS). This retrospective study included 143 patients who underwent MT for AIS between March 2021 and December 2023. Participants were stratified into two groups based on the presence of post-procedural HT: the HT group (n = 90) and the non-HT group (n = 53). Risk factors associated with the development of HT were examined using univariate and multivariate logistic regression analyses. A predictive model was subsequently constructed based on the independent risk factors identified. The discrimination, calibration, and clinical applicability of the nomogram model were evaluated using the receiver operating characteristic (ROC) curve with its area under the curve (AUC), calibration curve, and decision curve. Bootstrap resampling was performed 1,000 times to validate the model. The incidence of HT following MT was 62.9

ObjectiveTo obtain perioperative cerebral blood flow measurements and assess the role of intraoperative blood flow evaluation in predicting postoperative complications.MethodsWe conducted a retrospective analysis of 102 cases of extracranial-intracranial (EC-IC) bypass surgery performed for flow augmentation in patients with moyamoya disease. The study monitored blood flow in the donor artery, graft vessels, and recipient artery at various stages. We measured superficial temporal artery (STA) flow during the perioperative period using a transit time ultrasonic flowmeter and Doppler ultrasound for real-time monitoring. A modified MBC scale was developed to evaluate the vascular network of the middle cerebral artery (MCA). STA flow measurements and the modified MBC scale were analyzed for their correlation with the incidence of cerebral hyperperfusion syndrome (CHS).ResultsA total of 102 hemispheres underwent revascularization through EC-IC bypass, comprising 69 direct bypass cases and 33 combined bypass cases. We observed fluctuations in donor vascular flow during the perioperative period. The STA flow increased post-anastomosis, stabilizing at an elevated level thereafter. Specifically, the STA in situ flow measured 7.06 ± 3.30 mL/min, while the STA-Cut flow was 59.75 ± 37.49 mL/min. The flow following STA anastomosis was 36.07 ± 22.59 mL/min. Prior to the anastomosis of the bonnet aponeurosis and skin, STA flow was recorded at 35.14 ± 22.93 mL/min. Postoperative STA flow on day one was 111.91 ± 62.06 mL/min, and on day seven it was 104.47 ± 64.93 mL/min. Notably, 12.74% of patients developed CHS. Logistic regression analysis indicated that the occurrence of CHS was significantly correlated with STA-Cut flow, suggesting that lower flow rates are associated with a higher likelihood of CHS. Additionally, surgical approach and the MBC scale were also related to the incidence of CHS.ConclusionThe transit time flow (TTF) measurements indicate that the superficial temporal artery (STA) transitions from a low-flow vessel in situ to a medium- to high-flow graft vessel immediately after anastomosis. The variability in flow within the graft vessel plays a significant role in the occurrence of postoperative hyperperfusion syndrome.

Sepsis-associated encephalopathy (SAE) is a severe refractory neurological complication occurring in up to 70

Germinal matrix hemorrhage-intraventricular hemorrhage (GMH-IVH) is a severe complication frequently occurring in preterm infants, often resulting in permanent neurological impairment and persistent functional deficits. Human umbilical cord-derived mesenchymal stem cells have shown significant promise as a therapeutic approach in neonatal brain injury due to their robust paracrine effects, multipotent differentiation potential, and minimal immunogenic properties. The protective roles of human umbilical cord mesenchymal stem cells (hUC-MSCs) involve multiple synergistic pathways, such as suppressing neuroinflammatory responses, inducing apoptotic processes, stimulating neurogenesis and angiogenesis, and enhancing blood-brain barrier integrity. Additionally, hUC-MSCs and their extracellular vesicles (EVs) confer protective benefits through the secretion and delivery of bioactive substances, including cytokines and microRNAs (miRNAs), which can alleviate brain damage and subsequently enhance motor and cognitive outcomes. Although further large-scale clinical investigations are required to validate their effectiveness, current preclinical and animal studies provide preliminary evidence affirming the safety profile and therapeutic efficacy of hUC-MSCs treatment as an innovative strategy for managing GMH-IVH. LITERATURE SEARCH STRATEGY: We conducted a systematic literature search to identify relevant publications. The primary databases were PubMed and Web of Science, with the search period extending through October 2025. The search combined the following keywords and, where applicable, MeSH terms: "Germinal matrix hemorrhage," "Intraventricular hemorrhage," "GMH-IVH," "White matter injury," "Hydrocephalus," "mesenchymal stem cells," "MSC," "umbilical cord," "mechanism," "therapy," "neuroprotection," and "clinical trials." We also searched ClinicalTrials.gov for ongoing or completed trials of MSCs for neonatal brain injury. The selection prioritized original research, high-quality reviews, and meta-analyses. After an initial title-and-abstract screening, full texts of potentially relevant articles were examined. Studies offering the most significant insight into the mechanisms of hUC-MSC therapy in GMH-IVH were chosen for in-depth discussion in this review.

BackgroundCystic brain metastases (CBM) present significant clinical challenges due to their heterogeneity and the limitations of current diagnostic methods in guiding treatment. Traditional tissue biopsies are invasive and may not capture tumour heterogeneity, while plasma circulating tumour DNA (ctDNA) analysis is impeded by the blood-brain barrier, leading to low sensitivity for detecting intracranial lesions. These limitations create a critical gap in the personalised management of patients with CBM.MethodsWe evaluated the utility of cyst fluid ctDNA as a minimally invasive biomarker for genetic profiling and treatment monitoring in CBM patients. ctDNA was extracted from cyst fluid, tumour tissue, plasma, and cerebrospinal fluid (CSF) samples collected from 18 patients. NGS was performed to analyse genetic mutations. Mutation detection rates and genetic heterogeneity were compared across different sample types. Dynamic changes in ctDNA mutation abundance in cyst fluid were assessed in relation to treatment responses.ResultsCyst fluid ctDNA demonstrated a higher mutation detection rate and captured more significant genetic heterogeneity than plasma ctDNA and, in some cases, even matched tissue samples. Clinically significant mutations, including actionable driver genes such as EGFR and TP53, were identified in cyst fluid ctDNA but were undetectable in plasma. Moreover, dynamic changes in the abundance of ctDNA mutations in cyst fluid correlated with treatment responses, indicating its potential for real-time therapeutic efficacy monitoring.ConclusionsCyst fluid ctDNA provides a sensitive and comprehensive method for capturing the genetic landscape of CBM, effectively overcoming the limitations of tissue biopsies and plasma ctDNA analysis. By establishing a real-time molecular surveillance network, cyst fluid ctDNA analysis redefines precision neuro-oncology paradigms, transitioning CBM management from static histomolecular snapshots to adaptive therapeutic ecosystems.

WD Repeat Domain 4 (WDR4) is integral to the development and progression of various cancers; however, its specific role and underlying molecular mechanisms in glioma remain inadequately elucidated. This study undertook an analysis of WDR4 expression levels in glioma and normal brain tissues utilizing publicly accessible datasets from TCGA and GTEx project, with further validation conducted through the GEPIA and the HPA databases. Prognostic significance was assessed using Kaplan-Meier survival analysis and multivariate Cox regression models. Cellular functions were investigated through CCK-8 viability assays, colony formation assays, and cell cycle analysis, while the tumorigenic potential in vivo was corroborated using a nude mouse xenograft model. The findings revealed a significant upregulation of WDR4 in both glioma tissues and cell lines. Elevated WDR4 expression correlated with reduced overall survival and emerged as an independent prognostic factor. Functional assays indicated that WDR4 silencing markedly inhibited glioma cell proliferation, induced G1 phase cell cycle arrest, and resulted in the downregulation of CDK1 and CDK2 protein expression. Further co-expression analysis, GSEA, KEGG pathway enrichment, and western blotting suggested that WDR4 may exert its oncogenic effects through activation of the PI3K/Akt signaling pathway. In conclusion, WDR4 is highly expressed in glioma and promotes tumor progression via the PI3K/Akt-CDK1/2 signaling axis. These findings indicate that WDR4 may serve as a potential prognostic biomarker and therapeutic target in glioma.

Oxidative stress and neuronal apoptosis could be an important factor leading to post-hemorrhagic consequences after germinal matrix hemorrhage (GMH). Previously study have indicated that relaxin 2 receptor activation initiates anti-oxidative stress and anti-apoptosis in ischemia-reperfusion injury. However, whether relaxin 2 activation can attenuate oxidative stress and neuronal apoptosis after GMH remains unknown. To investigate the beneficial effect of relaxin 2 on oxidative stress injury and neuronal apoptosis by GMH, a total of 150 rat pups were subjected to GMH by an intraparenchymal injection of bacterial collagenase. Recombinant human relaxin-2 (rh-relaxin-2) was administered intraperitoneally injections at 1 h and 13 h after GMH. Lenti-virus with sgRXFP1 and sgCtrl was administered intracerebroventricular (i.c.v.) on the left side of the brain to inhibit the RXFP1 at 2d prior to GMH induction, and LY321499, ERK inhibitor, was administered by i.c.v. injection at 1 h on the left side of the brain prior to GMH induction, respectively. Co-immunoprecipitation, immunofluorescence, TUNEL, Fluoro-Jade C, DHE staining, western blot, Nitrix Oxide (NO) quantification and side effect experiments were performed to evaluate post-GMH. We found endogenous relaxin-2 interacts with RXFP1 and both protein colocalized in neurons on the first day after GMH. Additionally, RXFP1 activation with rh-relaxin-2 significantly inhibited oxidative stress and neuronal apoptosis in GMH + rh-relaxin-2 group compared with GMH + vehicle group. Moreover, rh-relaxin-2 treatment significantly inhibited the phosphorylation of ERK and nNOS, as well as upregulated expression of Bcl2 and NO and downregulated expression of Bax and Romo 1. The beneficial effects of rh-relaxin-2 were reversed by i.c.v. injection of lenti-virus with sgRXFP1 and LY321499, respectively. Furthermore, the side effect experiment showed rh-relaxin-2 did not affect neurological behavior and the function of liver and kidney. In conclusion, our finding showed that rh-relaxin-2 attenuated oxidative stress and neuronal apoptosis after GMH through RXFP1-ERK-nNOS-NO signaling pathway.

G protein-coupled receptors (GPCRs) play a pivotal role in maintaining the stemness of both normal and cancer stem cells. However, the function of the regulator of G protein signaling (RGS) family, particularly in tumor stem cells, remains poorly under-stood. Through bioinformatics analysis of clinical data, we identified RGS20 as a potential regulator of glioma stemness and temozolomide (TMZ) resistance, which may significantly influence patient prognosis. Subsequent in vitro and in vivo experiments demonstrated that RGS20 inhibition markedly enhanced tumor sphere formation and upregulated stem cell markers by intrinsically activating the WNT/β-catenin signaling pathway, thereby promoting tumorigenesis and ultimately leading to TMZ resistance. Furthermore, in human glioblastoma specimens, β-catenin signaling associated with low RGS20 expression was significantly enriched in hypoxic regions, suggesting that this mechanism may support the maintenance of glioma stem cells (GSCs) and drive TMZ resistance within the hypoxic niche. Our findings reveal that low RGS20 expression sustains WNT/β-catenin signaling in a ligand-reduced manner within hypoxic niches, unveiling a novel intracellular mechanism that drives glioma progression. Targeting this mechanism could provide new therapeutic strategies for glioma treatment.

Idiopathic pulmonary fibrosis (IPF) is a chronic progressive disease with poor survival, which is characterized by abnormal accumulation of fibrotic tissue in the lung parenchyma. Transforming growth factor-β1 (TGF-β) is a central profibrotic mediator, but the related mechanism of the activation of latent TGF-β has not been conclusively elucidated. A comprehensive study of mRNAs in human IPF was conducted using GSE10667 microarray data from GEO database, and the expression of cartilage intermediate layer protein (CILP) was upregulated among end-stage pulmonary fibrosis (EPF) and acute pulmonary fibrosis (APF) as compared to non-fibrosis tissues. Furthermore, CILP has protein-protein interactions with TGF-β1 through PPI analysis. Therefore, we investigated the potential effects and mechanisms of CILP in pulmonary fibrosis in pulmonary fibroblasts and BLM-induced mouse model (Eight-week-old male C57BL/6 mice, 20-22 g, purchased from the Experimental Animal Center of Guangzhou Medical University). In vitro, treatment with recombinant CILP (100 ng/mL) significantly attenuated TGF-β1-induced upregulation of collagen type I (Col1a1, p < 0.01) and α-smooth muscle actin (α-SMA, p < 0.01) in primary mouse pulmonary fibroblasts. Mechanistically, CILP suppressed TGF-β1-mediated SMAD3 phosphorylation (p-SMAD3, p < 0.001) and nuclear translocation, as confirmed by Western blotting and immunofluorescence. In the bleomycin (BLM)-induced mouse model of pulmonary fibrosis, intravenous administration of CILP (1 μg/g body weight, administered every 2 days for 4 weeks) reduced lung collagen deposition (Masson staining) by 38% (p < 0.01), lowered Ashcroft scores (from 5.8 ± 0.7 to 2.3 ± 0.4, p < 0.001), and decreased lung hydroxyproline content (a marker of collagen accumulation) by 42% (p < 0.01) compared to BLM-only controls. Clinically, serum CILP levels showed no significant difference between 17 idiopathic pulmonary fibrosis (IPF) patients and 17 non-fibrotic controls (3.2 ± 0.8 ng/mL vs. 3.5 ± 0.9 ng/mL, p > 0.05), suggesting potential lung tissue-specific action of CILP with minimal systemic off-target risk. In conclusion, CILP inhibited TGF-β1-induced fibrosis via its negative feedback loop, and may act as a promising candidate for the precaution and treatment of IPF.

OBJECTIVE:In adult patients with moyamoya disease (MMD), insufficient postoperative collateral formation is usually seen after combined bypass surgery. The arachnoid membrane, a critical anatomical barrier between extracranial and intracranial vascular systems, has an unclear role in postoperative angiogenesis. The aim of this study was to investigate whether intraoperative arachnoid membrane opening enhances angiogenesis in adult patients undergoing combined bypass surgery for MMD. METHODS:This multicenter, prospective, randomized controlled trial (Membrane Opening to Promote Angiogenesis in Adult MMD [MOPOAM]) was conducted across 8 institutions in China. Between June 2022 and August 2023, 104 patients diagnosed with MMD through digital subtraction angiography were randomly assigned to two groups: one group underwent arachnoid opening (n = 51), while the other group had arachnoid preservation (n = 53). The primary endpoint was the rate of good angiogenesis (Matsushima grade A and B) assessed 6-12 months postoperatively, and the secondary endpoint was the incidence of surgical complications within 1 month postsurgery. RESULTS:All 104 patients completed the study. The rate of good angiogenesis was 54.9% in the arachnoid opening group and 64.2% in the arachnoid preservation group, with no significant difference between groups (OR 1.470, 95% CI 0.669-3.229; p = 0.337). Postoperative complications were observed in 2 patients (3.9%) in the arachnoid opening group and in 5 patients (9.4%) in the arachnoid preservation group, showing no significant difference (OR 2.552, 95% CI, 0.472-13.796; p = 0.276). No significant association between arachnoid opening procedures and functional recovery (assessed by modified Rankin Scale scores) was found during the 6- to 12-month follow-up period (p = 0.831). CONCLUSIONS:In adults with MMD, arachnoid membrane opening during combined bypass surgery did not significantly improve postoperative angiogenesis rates or increase perioperative complications.

BACKGROUND:Colorectal cancer (CRC) ranks as the third most common cancer globally, with significant postoperative recurrence and metastasis rates. The heterogeneity of CRC presents challenges in the selection of adjuvant chemotherapy regimens, highlighting the need for personalized treatment strategies. The development of in vitro models for drug sensitivity testing, including a novel patient-derived tumor-like cell cluster (PTC) model, offers a potential solution for predicting drug efficacy and guiding treatment. METHODS AND DESIGN:This multicenter randomized controlled trial (RCT) aims to evaluate the consistency between the in vitro PTC drug sensitivity test results with whole exome sequencing and the clinical prognosis of CRC patients. The study will involve 200 patients who will be randomly assigned to receive either PTC-guided adjuvant chemotherapy or traditional chemotherapy. The primary endpoint is the 3-year disease-free survival rate (3yDFS), with secondary endpoints including the consistency between test results and clinical outcomes and the prognostic value of gene mutations and other biomarkers. DISCUSSION:This study represents a significant step toward precision medicine in CRC treatment by integrating PTC technology with whole-exome sequencing. These findings could provide valuable insights into personalized treatment approaches, potentially improving the clinical outcomes of patients with CRC. TRIAL REGISTRATION:ClinicalTrials.gov: NCT05424692. Registered on June 21, 2022.

Ferroptosis plays a crucial role in secondary brain injury following traumatic brain injury (TBI). Neutrophil extracellular traps (NETs) formation has also been implicated in secondary injury after TBI. However, the relationship between NETs, blood-brain barrier (BBB) disruption, and ferroptosis remains unclear. In this study, we found that levels of NET-related biomarkers in the plasma of TBI patients were significantly elevated and positively correlated with ICAM-1 and vWF. Moreover, inhibiting NET formation via peptidylarginine deiminase 4 (PAD4) deficiency reduced ferroptosis and BBB damage. In vitro experiments demonstrated that NETs exacerbated ferroptosis in hCMEC/D3 cells, leading to BBB disruption. Knockdown of ZBP1 and overexpression of FSP1 both reversed NETs-induced ferroptosis in endothelial cells and alleviated BBB damage. Additionally, we observed a close relationship between ZBP1 and FSP1. Collectively, our findings suggest that inhibiting NETs formation can mitigate ferroptosis and BBB damage, while targeting ZBP1 and FSP1 may offer potential therapeutic strategies for improving outcomes after TBI.

Abnormalities in the structure and function of neural circuits can disrupt normal physiological processes, leading to various disorders, such as Alzheimer's disease (AD) and Parkinson's disease (PD), which are closely associated with dysfunctions within brain neural circuits. Elucidating the architecture of these circuits is fundamental for understanding the mechanisms underlying information processing in the brain, facilitating investigations into the pathogenesis of diverse neurological diseases, and providing a theoretical foundation for their prevention, diagnosis, and treatment. Viral tracers have been instrumental in identifying novel neural circuits or revealing new properties of established circuits. Compared with small-molecule compounds and peptide tracers, neurotropic viruses offer several advantages as neural circuit tracers: first, they possess self-replicative capabilities that enable signal amplification; second, they can label both upstream and downstream neurons across synapses; and third, they are capable of delivering foreign genes into neuronal cells. Herpes simplex virus type 1 (HSV-1) stands out as a significant member among neurotropic viruses because of its unique neurotropic characteristics and transneuronal transmission properties, making it an ideal tool for tracing neural circuits.