BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) exert potent paracrine effects that can reshape the immune microenvironment. Nebulized inhalation enables non-invasive, targeted delivery to the lung, a pivotal immune interface capable of modulating systemic immunity. This study introduces a novel therapeutic strategy using nebulized BMSC supernatant to activate the FOXO3 signaling pathway in pulmonary dendritic cells (DCs), reprogramming them toward a tolerogenic phenotype. This approach suppresses autoreactive T cell infiltration and alleviates central nervous system (CNS) inflammation in the experimental autoimmune encephalomyelitis (EAE) model, offering a potential acellular therapy for multiple sclerosis (MS) and other autoimmune diseases. METHODS: EAE mice were treated with nebulized BMSC supernatant or adoptive transfer of pretreated DCs. Disease progression was assessed by body weight and clinical scores. Hematoxylin and eosin (HE) staining and myelin immunofluorescence staining were used to evaluate CNS inflammation and demyelination. Flow cytometry measured T cell differentiation in spleen and lymph nodes, as well as DC antigen presentation and cytokine secretion in lung tissue. Quantitative Polymerase Chain Reaction (QPCR) assessed inflammatory cytokines, and immunofluorescence determined FOXO3 expression in DCs. FOXO3 involvement was validated using inhibitor-pretreated DCs in adoptive transfer experiments. RESULTS: Nebulized BMSC supernatant reduced clinical severity and weight loss in EAE mice, and decreased CNS inflammatory infiltration and demyelination. Treatment suppressed peripheral Th1 and Th17 differentiation while increasing Treg frequency. Lung DCs exhibited reduced antigen presentation and pro-inflammatory cytokine expression, increased IL-10 secretion, and elevated FOXO3 expression. Blocking FOXO3 in DCs reversed these effects, aggravating EAE symptoms and promoting Th1/Th17 differentiation. CONCLUSIONS: Nebulized BMSC supernatant effectively treats EAE by inducing pulmonary tolerogenic dendritic cells (tolDC) via FOXO3 activation, reshaping peripheral T cell responses, and reducing CNS inflammation. This strategy highlights the therapeutic potential of targeting the pulmonary immune interface as a novel acellular approach for MS and related autoimmune disorders.

B-cell-depleting therapies have revolutionized multiple sclerosis (MS) treatment, yet relapses persist in some patients-suggesting additional pathogenic drivers beyond peripheral B cells. Tertiary lymphoid structures (TLS) are extensively documented in progressive MS at autopsy, but whether their formation begins during the relapsing-remitting phase and how they evolve during the transition to progression remain undefined. Here, using the relapsing-remitting PLP139-151-induced EAE model, we uncover that TLS-like structures form in the subventricular zone during relapse, once established, persist through remission as niches containing both B cells and persistently activated microglia. Neither B-cell depletion alone nor BTK inhibition alone fully prevents relapse. Strikingly, early combined B-cell depletion and BTK inhibition virtually abolishes TLS-like structure formation and may effectively prevent complete disease relapse in this model. By contrast, late initiation of the same combination fails to resolve existing TLS-like structures or prevent relapse, although it attenuates disease severity. These data indicate that established TLS-like structures may represent treatment-resistant compartments, and that both B cells and microglia may be crucial during early formation for sustaining their disease relapse-driving activity. Our study confirms that TLS-like structures may be a key factor driving the compartmentalization of central nervous system inflammation, points to a potentially narrow therapeutic window for intervention, and proposes that early combined B-cell depletion and BTK inhibition may represent a promising strategy worthy of further investigation.

Aging reshapes hematopoiesis toward myelopoiesis, particularly increased output of classical monocytes, contributing to immunosenescence. However, the underlying mechanisms remain unclear. Here, we identify that aging drives hematopoietic stem cells (HSCs) to progressively commit to a CCR2⁺ monocyte fate along a defined lineage trajectory, which is executed through a redox‐sensitive transcriptional cascade. We find that the age‐related decline in bone marrow IGF‐1 is the upstream niche signal that releases suppression of the oxidative stress/NF‐κB/IRF8 axis, allowing this cascade to redirect early progenitors toward a CCR2⁺ myeloid‐primed fate. Functional assays confirm that cells following this CCR2⁺ trajectory drive myeloid‐biased hematopoiesis and directly contribute to age‐associated immunosuppression. Our work delineates a hierarchical pathway from niche decay to lineage redirection, establishing the CCR2⁺ fate trajectory as a key driver of immunosenescence and a potential therapeutic target.

To examine whether pidotimod affects the progression and severity of experimental autoimmune encephalomyelitis (EAE), a classic animal model of multiple sclerosis (MS), the balance of splenic lymphocytes in pidotimod-treated and untreated EAE mice was examined. C57BL/6J mice were immunized by subcutaneous injection of an emulsion containing MOG35-55, with subsequent monitoring of their general condition and clinical scores following treatment with pidotimod or saline solution (vehicle control). Hematoxylin and eosin (H E) staining, along with flow cytometry (FCM), was employed to evaluate leukocyte infiltration, while FluoroMyelin™ Green staining was utilized to assess axonal demyelination in the central nervous system (CNS). Additionally, FCM was conducted to investigate the effects of pidotimod on splenic lymphocytes both in vitro and in vivo during the peak stage of EAE. Compared to the vehicle control, pidotimod treatment significantly reduced the clinical scores, decreased leukocyte infiltration in the spinal cord and brain, and suppressed demyelination in the spinal cord. Furthermore, pidotimod treatment markedly increased the populations of CD4+ CD25+ Foxp3+ regulatory T cells (Tregs) and CD8+ Foxp3+ Tregs, while decreasing the numbers of CD4+ IFN-γ+ helper T cells (Th1), CD4+ IL-17+ helper T cells (Th17), and CD8+ IL-17+ cytotoxic T cells (Tc17) in the spleen during the peak stage of EAE both in vitro and in vivo. Additionally, pidotimod treatment significantly diminished the population of B220+ TNF-α+ B cells in the spleen at the peak stage of EAE both in vitro and in vivo. The present study preliminarily explored the effects and potential immunomodulator mechanisms of pidotimod in treating EAE mice. Results indicated that pidotimod treatment decreased the percentages of CD4+ IFN-γ+ Th1 cells, CD4+ IL-17+ Th17 cells, CD8+ IL-17+ Tc17 cells and B220+ TNF-α+ B cells, while increasing the percentages of CD4+ CD25+ Foxp3+ Tregs and CD8+ Foxp3+ Tregs in the spleen at the peak stage of EAE. Additionally, pidotimod reduced leukocyte infiltration into the spinal cord and brain, as well as demyelination in the spinal cord. These findings suggest that the neuroprotective effects of pidotimod in EAE mice may be its ability to regulate the balance of splenic lymphocytes.

AIMS:Effective remyelination in multiple sclerosis (MS) requires both the proliferation of endogenous neural stem cells (NSCs) and their lineage-specific differentiation into oligodendrocyte progenitor cells (OPCs). This study aimed to investigate whether electroacupuncture (EA) promoted NSC proliferation and OPC differentiation via β-endorphin (β-EP)-mediated opioid signaling in a murine model of MS. METHODS:Experimental autoimmune encephalomyelitis (EAE) was induced in C57BL/6 mice to model MS. EA stimulation was applied daily at the Zusanli (ST36) acupoint. NSC proliferation and OPC differentiation were assessed via immunofluorescence, flow cytometry (FCM), and RT-qPCR. β-EP expression and opioid receptor involvement were evaluated in the hypothalamus and subventricular zone (SVZ). Naloxone, a nonselective opioid receptor antagonist, was used to determine the role of opioid signaling in EA-induced effects. RESULTS:EA significantly enhanced NSC proliferation and increased the proportion of NSC-derived OPCs in the SVZ of EAE mice. EA treatment improved clinical score, reduced demyelination, and attenuated leukocyte infiltration of the central nervous system (CNS). Mechanistically, EA upregulated β-EP and its precursor pro-opiomelanocortin (POMC), along with opioid receptors μ-opioid receptor (MOR) and κ-opioid receptor (KOR) (encoded by Oprm1 and Oprk1, respectively). Naloxone administration abolished the beneficial effects of EA on NSC behavior and remyelination, confirming the involvement of opioid receptor-dependent β-EP signaling. CONCLUSION:EA promotes remyelination in EAE mice by stimulating β-EP-mediated NSC proliferation and OPC differentiation. These findings reveal a novel neuroregenerative mechanism and support EA as a promising adjunctive strategy for demyelinating diseases such as MS.

It has been reported that the nervous system can regulate immune reactions through various mechanisms. However, the role of splenic sympathetic nerve activity in the autoimmune reactions during the pathogenesis of experimental autoimmune encephalomyelitis (EAE) remained unclear. Here, we blocked the activity of the splenic sympathetic nerve and found that the number of adaptive immune cells, such as CD4+ T cells, CD8+ T cells and B cells, were upregulated. Additionally, there was an increase in the secretion of inflammatory cytokines in the spleen, and the neurological symptoms of EAE were exacerbated. In vitro experiments, we found that norepinephrine (NE), the neurotransmitter of the splenic sympathetic nerve, indirectly drove the death of effector CD8+ T cells. Furthermore, activated B cells, under the influence of NE, specifically recognized effector CD8+ T cells by upregulating MHC-I molecules and killed these cells via the FasL/Fas pathway. Our findings provide a new perspective on B cells killing effect in vitro, which was boosted by NE and demonstrate that the splenic sympathetic nerve controls the degree of autoimmune responses in EAE This adds a new dimension to the diversity of NE’s regulatory effects on adaptive immune cells and suggests a potential new therapeutic approach for autoimmune diseases.

C-terminal binding protein-2 (Ctbp2) is an evolutionarily conserved transcriptional repressor that regulates fundamental processes such as cell proliferation and apoptosis. However, the potential role of Ctbp2 in cardiomyocyte proliferation and heart regeneration remains unclear. In this study, we aim to explore the important role of Ctbp2 in cardiomyocyte proliferation and the regeneration of injured adult hearts. In this study, we found that the expression of Ctbp2 in cardiomyocytes is downregulated after adulthood. Silencing Ctbp2 in cardiomyocytes on the post-natal day 1 (P1) reduced the proliferation ability of cardiomyocytes, whereas overexpressing Ctbp2 enhanced the proliferation ability of cardiomyocytes. Additionally, overexpressing Ctbp2 via adeno-associated virus-9 (AAV9) had no effect on the hearts of normal adult mice, but in the case of heart injury, overexpression of Ctbp2 in adult mice cardiomyocytes promoted cardiomyocyte proliferation. Mechanistically, the transcriptional repressor Ctbp2 acts as a metabolite sensor, and its regulation of cardiomyocyte proliferation is influenced by the metabolites NADH/NAD+ and fatty acyl-CoAs. Ctbp2 is activated by the intracellular accumulation of NADH during cardiomyocyte ischemia and hypoxia, inhibiting the transcriptional activity of the transcription factor FoxO1, thereby repressing the expression of the target genes and cell cycle negative regulators p21 and p27, allowing cardiomyocytes to re-enter the cell cycle. In contrast, normal adult cardiomyocytes mainly use fatty acid oxidation metabolism as their primary energy source, and the intracellular production of fatty acyl-CoAs inactivates Ctbp2, thus preventing it from inhibiting FoxO1 mediated cell cycle arrest. In conclusion, this study demonstrates that the Ctbp2-FoxO1-p21/p27 axis can promote cardiomyocyte proliferation and heart regeneration. As a metabolite sensor, Ctbp2 is activated during cardiomyocyte ischemia and hypoxia, while it is inactivated under normal conditions. This controllable and transient regulation of cardiomyocyte proliferation can avoid the detrimental effects on cardiac function caused by long-term regulation of cardiomyocyte proliferation, such as hypertrophic cardiomyopathy or heart failure. This provides new targets and new ideas for addressing the issues of cardiomyocyte proliferation and heart regeneration.

This study identified 13 endoplasmic reticulum stress (ERS)-related biomarkers associated with multiple sclerosis (MS) through integrated bioinformatics analysis (including weighted gene co-expression network analysis and machine learning algorithms) and single-cell sequencing, combined with validation in an experimental autoimmune encephalomyelitis (EAE) mouse model. Among them, GPX1, RCN1, and UBE2D3 exhibited high diagnostic value (AUC > 0.7, p < 0.05), and the diagnostic potential of GPX1 and RCN1 was confirmed in the animal model. The study found that memory B cells, plasma cells, neutrophils, and M1 macrophages were significantly increased in MS patients, while naive B cells and activated NK cells decreased. Consensus clustering based on key ERS-related genes divided MS patients into two subtypes. Single-cell sequencing showed that microglia and pericytes were the cell types with the highest expression of key ERS-related genes, and the APP-CD74 pathway was enhanced in the brain tissue of MS patients. Mendelian randomization analysis suggested that GPX1 plays a protective role in MS. These findings reveal the mechanisms of ERS-related biomarkers in MS and provide potential targets for diagnosis and treatment.

Evobrutinib, a third-generation Bruton's tyrosine kinase (BTK) inhibitor, shows great promise for treating neuroinflammatory diseases due to its small molecular size, ease of absorption, and ability to cross the blood–brain barrier. Although previous studies have confirmed significant BTK expression in microglia, the potential of Evobrutinib to treat ischemic stroke by modulating microglial function and its underlying mechanisms remain to be elucidated. Male C57BL/6 mice with cerebral ischemia was established to evaluate the effects of oral Evobrutinib treatment. Assessments included TTC staining, behavioral experiments, and pathological examinations were used to evaluate cerebral ischemic injury. Western Blot, flow cytometry, and qPCR were employed to monitor changes in BTK and pBTK expression in microglia and the impact of Evobrutinib on neuroinflammation following the stroke. In vitro, primary microglia were generated to determine the effects of Evobrutinib on the TLR4/ Myd88/NF-κB pathway and on the polarization of microglial subtypes. The expression of BTK and pBTK is upregulated in microglia under conditions of cerebral ischemia and oxygen–glucose deprivation (OGD). Evobrutinib treatment not only reduced infarct volume in mice but also ameliorated pathological damage and facilitated neurological function recovery. Flow cytometry revealed that Evobrutinib decreased inflammatory cell infiltration and promoted M2 microglia polarization post-stroke. In vitro studies demonstrated that Evobrutinib downregulated the proportion of pro-inflammatory microglia and curtailed the secretion of inflammatory factors under OGD conditions. Mechanistically, Evobrutinib attenuated the OGD-induced upregulation of TLR4/Myd88/NF-κB expression, an effect that was further enhanced by the addition of the TLR4 pathway inhibitor TAK242. Evobrutinib inhibits the expression and activation of BTK in microglia, reducing M1 microglia-mediated neuroinflammation and alleviating ischemic injury following stroke. This effect is mechanistically linked to the inhibition of TLR4/Myd88/NF-κB-mediated M1 polarization of microglia. Evobrutinib treatment improves neurological function of mice with cerebral ischemia, and alleviates neuroinflammation by inhibiting M1 microglia polarization through TLR4/Myd88/NF-κB pathway.

Background:Imaging, particularly multimodal magnetic resonance imaging (MRI), serves as an essential auxiliary examination for diagnosing autoimmune encephalitis (AE). The diversity of autoantibodies complicates the imaging presentation of AE, exhibiting both common and individual features across different subtypes of AE. Currently, there is a lack of comprehensive studies on the imaging features of different subtypes of AE. The study aimed to explore imaging biomarkers for AE mediated by various subtypes of antibodies and clarify their significance in disease severity, treatment response, and prognosis. Methods:The clinical and imaging data of 45 patients with AE at The First Affiliated Hospital of Dalian Medical University, collected from January 2013 to August 2022, were analyzed. Patients underwent multi-modal brain MRI. Lesion probability maps were generated, and regions of interest (ROIs) were selected based on lesion location and clinical-electroencephalographic features, for measurement of three-dimensional T1-weighted imaging (3D-T1WI), T2-weighted imaging (T2WI), T2 fluid-attenuated inversion recovery (T2 FLAIR), and apparent diffusion coefficient (ADC) sequences. These values were used for correlating with disease severity, antibody titers, response to treatment, and prognosis. Results:The study included 45 AE patients: 18 with anti-leucine-rich glioma inactivated protein 1 (anti-LGI1), 11 with anti-N-methyl-D-aspartate receptor (anti-NMDAR), 5 with anti-gamma-aminobutyric acid receptor B (anti-GABABR), 4 with anti-myelin oligodendrocyte glycoprotein (MOG), 4 with anti-glutamate decarboxylase 65 (anti-GAD65), and 3 with anti-contactin-associated protein-like 2 (anti-Caspr2) encephalitis. MRI abnormalities were present in 62.2% of patients, lower than that of electroencephalography (EEG) (95.6%, P<0.05). Imaging typically showed common features across different AE subtypes, predominantly involving the limbic system or regions outside of it, manifesting as T1 hypointensity, T2 FLAIR hyperintensity or mild hyperintensity, and normal or mild hyperintensity on diffusion-weighted imaging (DWI). Different AE subtypes displayed specific imaging features: anti-LGI1 encephalitis often involved 2 locations: unilateral or bilateral hippocampus or basal ganglia; anti-NMDAR encephalitis showed a low rate of imaging abnormalities, with diffuse and unfixed cortical or subcortical T2 FLAIR hyperintensity. Anti-GABABR encephalitis primarily affected the temporal lobe or hippocampus. MOG antibody cortical encephalitis exhibited cortical swelling with T2 FLAIR hyperintensity in unilateral or bilateral hemispheres, particularly in the frontal lobe. Anti-GAD65 encephalitis involved the temporal lobe/hippocampus or pontocerebellar regions. The ADC value within the ROI positively correlated with both disease severity (r=0.6891, P<0.0001) and prognosis score (r=0.8102, P<0.0001). Further analysis using receiver operating characteristic (ROC) curve and binary logistic regression indicated that the ADC value was a risk factor for poor prognosis. Conclusions:Imaging abnormalities are less frequent than those detected by EEG but exhibit distinct features by subtype. Functional imaging enhances diagnostic accuracy. ADC values can serve as a crucial prognostic indicator.

RATIONALE AND OBJECTIVES:Glioblastoma (GBM) and solitary brain metastasis (SBM) exhibit similar radiomics features on magnetic resonance imaging (MRI), yet their treatment strategies and prognoses significantly differ. Therefore, accurate differentiation between these two types of tumors is crucial for clinical decision-making. This study aims to establish and validate an efficient diagnostic model based on the radiomic features of the T1-weighted contrast-enhanced (T1CE) sequence in the 10 mm brain-tumor interface region to achieve precise differentiation between GBM and SBM. METHODS:This study retrospectively collected contrast-enhanced T1-weighted imaging data from 226 GBM patients and 206 SBM patients at three centers between January 2010 and October 2024. Samples from centers 1 and 2 were used as the training set, while samples from center 3 were used as the test set. Two observers manually delineated the tumor edges on the T1CE images layer by layer to obtain the Region of Interest (ROI) covering the entire tumor volume. A 10 mm brain-to-tumor interface (BTI) was extracted using Python code. Radiomic features were extracted from the 10 mm BTI region, followed by feature selection and model construction. Finally, SHAP (SHapley Additive exPlanations) was used to visualize the model. Three radiologists with 2, 6, and 18 years of diagnostic experience independently evaluated the test set samples without knowing the patient information or pathology results, establishing three diagnostic models. The DeLong test was used to compare these models with the radiomic model. RESULTS:Ultimately, ten radiomic features were used for modeling. The model established using the logistic regression (LR) algorithm had an AUC of 0.893 on the training set and 0.808 on the test set. The AUCs of the three radiologists with different diagnostic experiences on the test set were 0.699, 0.740, and 0.789, respectively, all lower than that of the radiomic model. The DeLong test showed that ModelBTI performed significantly better than Doctor 1 (p<0.05) in the test set, but there was no statistically significant difference in performance between ModelBTI and Doctors 2 and 3. CONCLUSION:The radiomic model constructed based on the 10 mm brain-tumor interface can effectively differentiate between GBM and SBM, capturing tumor heterogeneity from a new perspective, thereby significantly improving diagnostic performance and providing assistance for clinical diagnosis. DATA AVAILABILITY STATEMENT:The original contributions presented in the study are included in the article/Supplemental material, further inquiries can be directed to the corresponding authors.

BACKGROUND:This study aims to explore the value of habitat-based magnetic resonance imaging (MRI) radiomics for predicting the origin of brain metastasis (BM). PURPOSE:To investigate whether habitat-based radiomics can identify the metastatic tumor type of BM and whether an imaging-based model that integrates the volume of peritumoral edema (VPE) can enhance predictive performance. METHODS:A primary cohort was developed with 384 patients from two centers, which comprises 734 BM lesions. An independent cohort was developed with 28 patients from a third center, which comprises 70 BM lesions. All patients underwent T1-weighted contrast-enhanced (T1CE) and T2-weighted (T2W) MRI scans before treatment. Radiomics features were extracted from tumor active area (TAA) and peritumoral edema area (PEA) selected using the least absolute shrinkage and selection operator (LASSO) to construct radiomics signatures (Rads). The Rads were further integrated with VPE to build combined models for predicting the metastatic type of BM. Performance of the models were assessed through receiver operating characteristic (ROC) curve analysis. RESULTS:Rads derived from TAA and PEA both showed predictive power for identifying the origin of BM. The developed combined models generated the best performance in the training (AUCs, lung cancer [LC]/non-lung cancer [NLC] vs. small cell lung cancer [SCLC]/non-small cell lung cancer [NSCLC] vs. breast cancer [BC]/gastrointestinal cancer [GIC], 0.870 vs. 0.946 vs. 0.886), internal validation (area under the receiver operating characteristic curves [AUCs], LC/NLC vs. SCLC/NSCLC vs. BC/GIC, 0.786 vs. 0.863 vs. 0.836) and external validation (AUCs, LC /NLC vs. SCLC/NSCLC vs. BC/GIC, 0.805 vs. 0.877 vs. 0.774) cohort. CONCLUSIONS:The developed habitat-based radiomics models can effectively identify the metastatic tumor type of BM and may be considered as a potential preoperative basis for timely treatment planning.

Astrocytes are key regulators of neuroinflammation in multiple sclerosis (MS). Electroacupuncture (EA), a safe and cost-effective adjuvant therapy, has shown benefits in neurodegenerative diseases, but its astrocyte-related mechanisms remain unclear. Here, we demonstrated that EA at ST36 alleviated blood-brain barrier (BBB) disruption and neuroinflammation during the peak period of experimental autoimmune encephalomyelitis (EAE). Additionally, EA at ST36 upregulated the expression of α-melanocyte-stimulating hormone (α-MSH) and its receptor melanocortin-4 receptor (MC4R) in spinal astrocytes. Pharmacological studies showed that MC4R agonist RO27-3225 mimicked the therapeutic effects of EA, whereas MC4R antagonist TCMCB07 weakened EA-mediated BBB protection and neuroinflammation suppression. Moreover, astrocyte-specific silencing of MC4R via adeno-associated virus (AAV) weakened EA-mediated BBB protection and neuroinflammation suppression. RNA-sequencing (RNA-seq) and western blot (WB) revealed that EA exerts neuroprotective effects by activating MC4R to inhibit MAPK and NF-κB signaling pathways. Moreover, in MC4R-overexpressing astrocytes, α-MSH and RO27-3225 reduced inflammation responses, while TCMCB07 reversed the effects by MAPK/NF-κB signaling pathways. Collectively, our findings identify astrocytic MC4R as a critical mediator of EA-driven neuroprotection by suppressing MAPK/NF-κB signaling, providing mechanistic insight and a promising therapeutic target for EAE and other neuroinflammatory disorders.

T cell dysfunction is a pivotal driving factor in autoimmune diseases, yet its underlying regulatory mechanisms remain incompletely understood. The role of long non-coding RNAs (lncRNAs) in immune regulation has gradually been recognized, although their functional mechanisms in T cells remain elusive. This study focuses on lncBADR (LncRNA Branched-chain Amino acids Degradation Regulator), elucidating its mechanism by which it regulates branched-chain amino acids (BCAAs) metabolism to influence T cell effector functions. Mice with specific knockout of lncBADR (T celllncBADR−/−) exhibited markedly ameliorated experimental autoimmune encephalomyelitis (EAE) symptoms. Mechanistic investigations revealed that lncBADR inhibits BCAAs degradation by binding to the enzymes Mccc1 and Pcca, leading to the accumulation of BCAAs within T-cells. This, in turn, activates the mTOR-Stat1 signaling pathway, promoting IFN-γ secretion and exacerbating EAE pathology. In contrast, knockout of lncBADR restored BCAAs degradation, significantly reducing IFN-γ secretion in T cells and suppressing their pathogenic functions. Further studies demonstrated that high-BCAAs feeding partially reversed the protective effects of lncBADR knockout, indicating that lncBADR plays a crucial role in autoimmune inflammation by regulating BCAAs metabolism. This study offers new insights into targeting lncBADR or modulating BCAAs metabolism as potential therapeutic strategies for autoimmune diseases.

Objective: The aim of this study is to elucidate the role of M2 macrophages in the pathogenesis of benign airway stenosis using a Sprague Dawley (SD) rat model and in vitro macrophage-fibroblast co-culture systems. Methods: Ligand-receptor interactions mediating cellular crosstalk between macrophages and fibroblasts were identified through single-cell RNA sequencing-based bioinformatics analysis. An airway stenosis model was established in SD rats, which were assigned to five experimental groups: normal control and post-modeling days 1 (D1), 4 (D4), 7 (D7), and 14 (D14). Temporal changes in M2 macrophage infiltration and their involvement in airway remodeling were assessed. Fibroblasts isolated from human granulation tissue and normal airway tissue were evaluated for differential activation of intracellular signaling pathways. In vitro macrophage-fibroblast co-culture systems involving M2 macrophages and fibroblasts were conducted to assess molecular signaling interactions. Results: A progressive increase in M2 macrophage infiltration was observed during the development of airway stenosis, accompanied by upregulation of secreted phosphoprotein-1 (SPP1) and activation of the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) signaling pathway. Fibroblasts derived from granulation tissue exhibited higher levels of pathway activation compared to normal fibroblasts.In co-culture, M2 macrophages induced fibroblast activation and fibrogenesis via SPP1-mediated signaling. Administration of rapamycin, an mTOR pathway inhibitor, significantly reduced granulation tissue formation and improved airway patency in the rat model. Conclusion: M2 macrophages contribute to fibrotic airway remodeling in benign airway stenosis through SPP1-mediated activation of the PI3K/AKT/mTOR signaling pathway in fibroblasts. Pharmacological targeting of this axis with rapamycin may represent a potential therapeutic strategy for mitigating fibrosis in benign airway stenosis.

OBJECTIVE:To clarify therapeutic potential and underlying mechanism of L-arginine (L-arg) in intervertebral disc degeneration (IDD). DESIGN:Human nucleus pulposus (NP) samples (n=36) underwent metabolomics, L-arg quantification, and immunohistochemistry to characterize the relationship between L-arg metabolism and IDD severity. In vitro NP cell experiments integrated RNA-sequencing, Western blot, co-immunoprecipitation, and immunofluorescence colocalization to clarify the therapeutic effects and mechanism of L-arg. In vivo validation using rat models (n=30) was performed with magnetic resonance imaging (MRI) and immunohistochemistry to assess the efficacy of L-arg in alleviating IDD. RESULTS:Metabolomics revealed a depletion of L-arg in severely degenerated NP tissues. L-arg supplementation mitigated TNFα-induced cellular senescence (β-galactosidase positive rate reduced by 37.4%, 95% CI: 29.5-45.3), extracellular matrix dysfunction (ACAN increased 1.59-fold, 95% CI: 1.19-2.11), and mitochondrial dysfunction (ATP content increased 1.43-fold, 95% CI: 1.29-1.57) in NP cells by inhibiting mitochondrial fragmentation. In rat IDD models, L-arg attenuated disc degeneration (MRI intensity: 3.245-fold vs. degeneration, 95% CI: 2.05-5.13). Mechanistically, L-arg suppressed inflammation-driven Tribbles homolog 3 (TRIB3) expression (0.49-fold vs. TNFα, 95% CI: 0.29-0.84). Elevated TRIB3 disrupted the interaction between A-kinase anchoring protein 1 (AKAP1) and protein kinase A regulatory subunit IIα (PKA RIIα)-an interaction critical for Drp1-S656 phosphorylation that inhibits mitochondrial fission. By preserving the AKAP1-PKA RIIα interaction, L-arg sustained Drp1 phosphorylation at S656 and blocked pathological mitochondrial fission. CONCLUSION:L-arg may alleviate IDD in vitro and vivo by modulating TRIB3-AKAP1-PKA/Drp1(S656) axis, highlighting its promising therapeutic potential in IDD.

Multiple sclerosis (MS) is a progressive autoimmune disease characterized by massive inflammatory infiltration, demyelination, and subsequent axonal injury and neuronal damage in the central nervous system (CNS). The etiology of MS remains unclear and there is not yet a definitive therapeutic schedule for the disease. Bone marrow mesenchymal stem cells (BMSCs), exhibiting neuroimmune-modulatory functions to alleviate various autoimmune diseases, show great potential in the treatment of MS. However, the instability of BMSCs-mediated immunosuppression in vivo has limited their application. MiR181-a, a positive regulator of immune balance, which has a preference for T cells and B cells differentiation, but degrade rapidly upon entering systemic circulation due to their unstable molecular structure. We propose a synergistic therapy approach that combines the penetrative targeting capability of BMSCs with the immuno-modulatory effects of miR181-a by overexpressing miR181-a to BMSCs through lentivirus packaging system. With this strategy, on the basis of the establishment of the experimental autoimmune encephalomyelitis (EAE) model, miR181-a overexpressing BMSCs (miR181a-BMSCs) would have a stronger immuno-modulatory treatment benefit, in terms of attenuating MS development. Indicate that this method prolongs the modulatory effects of BMSCs and resulted in significantly enhancements of the proliferation of regulatory B cells (Bregs), regulatory T cells (Tregs) and the inhibition of Th17 cells compared to the traditional BMSCs group. Moreover, 10-fold miRNA’s concentration in the exosome of miR181a-BMSCs, leading to an increased duration of miRNAs to exert their biological effects. By immunotherapy and synergistic treatment, the effectiveness of the treatment is significantly enhanced, showing consistent results in different groups of the animal model. This strategy takes advantage of BMSCs and miRNA and thus presents an effective synergistic strategy for the treatment of autoimmune diseases.

ObjectiveThe purpose of this study is to explore whether machine learning can be used to establish an effective model for the diagnosis of Parkinson's disease (PD) by using texture features extracted from cerebellar gray matter and white matter, so as to identify subtle changes that cannot be observed by the naked eye.MethodThis study involved a data collection period from June 2010 to March 2023, including 374 subjects from two cohorts. The Parkinson's Progression Markers Initiative (PPMI) served as the training set, with control group and PD patients (HC: 102 and PD: 102) from 24 global sites. Our institution's data was utilized as the test set (HC: 91 and PD: 79). Machine learning was employed to establish multiple models for PD diagnosis based on texture features of the cerebellum's gray and white matter. Results underwent evaluation through 5-fold cross-validation analysis, calculating the area under the receiver operating characteristic curve (AUC) for each model. The performance of each model was compared using the Delong test, and the interpretability of the optimized model was further augmented by employing Shapley additive explanations (SHAP).ResultsThe AUCs for all pipelines in the validation dataset were compared using FeAture Explorer (FAE) software. Among the models established by Kruskal-Wallis (KW) and logistic regression via Lasso (LRLasso), the AUC was highest using the “one-standard error” rule. 'WM_original_glrlm_GrayLevelNonUniformity' was considered the most stable and predictive feature.ConclusionThe texture features of cerebellar gray matter and white matter combined with machine learning may have potential value in the diagnosis of Parkinson's disease, in which the heterogeneity of white matter may be a more valuable imaging marker.

Ischemic heart disease invariably leads to devastating damage to human health. Nicotinamide ribose (NR), as one of the precursors of NAD+ synthesis, has been discovered to exert a protective role in various neurological and cardiovascular disorders. Our findings demonstrated that pretreatment with 200 mg/kg NR for 3 h significantly reduced myocardial infarct area, decreased levels of CK-MB and LDH in serum, and improved cardiac function in the rats during myocardial ischemia-reperfusion (I/R) injury. Meanwhile, 0.5 mM NR also effectively increased the viability and decreased the LDH release of H9c2 cells during OGD/R. We had provided evidence that NR pretreatment could decrease mitochondrial reactive oxygen species (mtROS) production and MDA content, and enhance SOD activity, thereby mitigating mitochondrial damage and inhibiting apoptosis during myocardial I/R injury. Further investigations revealed that NR increased NAD+ content and upregulated SIRT3 protein expression in myocardium. Through using of SIRT3 small interfering RNA and the SIRT3 deacetylase activity inhibitor 3-TYP, we had confirmed that the cardioprotective effect of NR on cardiomyocytes was largely dependent on the inhibition of mitochondrial oxidative stress via SIRT3-SOD2 axis. Overall, our study suggested that exogenous supplementation with NR mitigated mitochondrial damage and inhibited apoptosis during myocardial I/R injury by reducing mitochondrial oxidative stress via SIRT3-SOD2-mtROS pathway.

BackgroundMultiple sclerosis (MS) is a chronic, progressive autoimmune disease, with increasing attention on the role of B cells in its pathogenesis. Despite this growing interest, a comprehensive analysis of research trends and emerging foci on B cells in MS is currently lacking. In this research, we utilize a bibliometric approach to visualize and analyze research trends and focal points in this field, offering a valuable reference for future mechanistic studies in MS.MethodsWe retrieved bibliometric data from the Web of Science Core Collection (WOSCC) for articles published between 2014 and 2023. VOSviewer 1.6.18 and CiteSpace 5.7R3 were used for co-authorship, co-occurrence, and citation analyses to identify key researchers, institutions, countries, and emerging themes in B cell research related to MS.ResultsThe analysis examined 5,578 articles published in 1,041 journals by 5,337 institutions globally. The United States leads in publication output, with Amit Bar-Or identified as the most influential author, and Frontiers in Immunology as the top journal in the field. Research has increasingly focused on the complex role of B cells in MS, particularly their involvement in the central nervous system (CNS) and mechanisms of anti-B cell therapy. Recent trends point to a growing focus on meningeal inflammation, kinase inhibitors, and Epstein-Barr virus, signaling a shift in research priorities.ConclusionThis bibliometric analysis highlights pivotal research trends, key contributors, and emerging areas of interest in B cell research in MS from 2013 to 2024. The findings underscore the growing recognition of the multifaceted role of B cells in MS pathogenesis, particularly their involvement in the CNS compartment and the potential of targeted therapies. The study identifies meningeal inflammation, Epstein-Barr virus infection, and kinase inhibitors as promising avenues for future research. The analyses driving the in-depth exploration of B cell mechanisms in MS and the development of novel diagnostic and therapeutic strategies provide researchers in the MS field with a comprehensive and objective perspective, serving as a valuable reference for accelerating the translation of basic research findings into clinical applications.