Acute kidney injury (AKI) is a common and serious complication in the intensive care unit (ICU), and continuous renal replacement therapy (CRRT) is an important treatment option. However, there is no clear standard for the optimal time to stop CRRT. The cardiorenal interaction effect suggests that there may be a potential link between cardiac function and CRRT. This study aimed to investigate the left ventricular diastolic function for predicting CRRT discontinuation. This is a prospective cohort study. The study included 154 AKI patients admitted to ICUs undergoing CRRT from February 2023 to August 2024, which were divided into the successful downtime group (87 patients) and the failure group (67 patients), and their baseline data, laboratory indices, and ultrasound parameters were analyzed. The results showed that serum creatinine, urea nitrogen, and E/eʹ measured at the time of CRRT discontinuation were significantly lower in the successful group than in the unsuccessful group (all P < 0.05). A higher E/eʹ measured at the time of CRRT discontinuation was associated with a lower likelihood of successful CRRT discontinuation (OR 0.71, 95

Background and ObjectivesInebilizumab and rituximab (RTX) are anti-CD19 and anti-CD20 B cell-depleting antibodies, respectively. They are both used in the treatment of neuromyelitis optica spectrum disorders (NMOSD). However, limited data on both drugs exist regarding real-world clinical applications. This study aimed to assess the efficacy and safety of inebilizumab vs low-dose RTX in NMOSD patients through a retrospective-prospective multicenter analysis.MethodsThis study collected data on aquaporin-4 immunoglobulin G seropositive NMOSD patients from 6 cities in China among those receiving either inebilizumab or low-dose RTX (500 mg), with a 1-year follow-up. Analyses assessed outcomes using inverse probability of treatment weighting and doubly robust models.ResultsWe included a total of 229 patients, comprising 119 treated with inebilizumab and 110 with low-dose RTX. The cohort was predominantly female (217/229, 94.76%). The median follow-up duration was 12.0 months (range: 4.0-12.0) in the inebilizumab group and 12.0 months (range: 7.0-12.0) in the low-dose RTX group. Relapses occurred in 8/119 (6.72%) inebilizumab recipients vs 24/110 (21.82%) low-dose RTX recipients (hazard ratio [HR] 3.77, 95% confidence interval [CI] 1.56-9.14; p = 0.003). Adjusted annualized relapse rates were significantly lower in the inebilizumab group (0.06) than in the low-dose RTX group (0.24), corresponding to an incidence rate ratio (IRR) of 3.65 (95% CI 1.59-8.39; p = 0.003). Adverse events (AEs) occurred in 35 patients (29.42%) with inebilizumab, including 2 serious adverse events (SAEs; 1.68%), vs 44 patients (40.00%) with low-dose RTX (4 SAEs [3.64%]). Both AE and SAE rates showed no statistical difference between groups. The rate of patients with at least 1 infection in inebilizumab group was lower than that in the low-dose RTX group (p = 0.003).DiscussionIn this short-term study, inebilizumab demonstrated greater efficacy in reducing relapse risk, along with significantly lower rates of key adverse events, compared with low-dose RTX in patients with NMOSD. These findings support the use of inebilizumab as an effective and well-tolerated therapeutic option in a broader NMOSD population.Classification of EvidenceThis short-term study with a 12-month observation period provides Class III evidence that in patients with NMOSD, inebilizumab is more effective than low-dose RTX in reducing relapses.

BACKGROUND:The treatment of non-small cell lung cancer (NSCLC) has challenges such as drug resistance and recurrence. Concurrently, the induction of apoptosis and ferroptosis is a promising therapeutic strategy. This study aimed to investigate whether the natural product, verbascoside, induces apoptosis and ferroptosis in NSCLC cells by targeting BCAT2. METHODS:Bioinformatic analysis was used to predict the potential targets of verbascosides. Stable cell lines with BCAT2 knockdown and overexpression were constructed. The effects of verbascoside on NSCLC were evaluated in vitro and using a mouse xenograft model. RESULTS:Bioinformatics screening and molecular docking identified BCAT2 as a potential target of verbascoside, with a significantly stronger binding energy (-7.8 kcal/mol) than another candidate, PARP1. In vitro and in vivo experiments confirmed that BCAT2 knockdown significantly inhibited NSCLC cell viability, induced apoptosis and ferroptosis, and induced mitochondrial damage. Conversely, BCAT2 overexpression produced opposite effects. Verbascoside treatment inhibited BCAT2 expression in a concentration-dependent manner, recapitulating the apoptotic, ferroptotic, and mitochondrial damage phenotypes induced by BCAT2 knockdown; however, BCAT2 overexpression significantly reversed these effects of verbascoside. In an animal model, treatment with verbascoside significantly suppressed tumor growth and activated apoptosis and ferroptosis in tumor tissues by downregulating BCAT2. CONCLUSIONS:Verbascoside can induce apoptosis and ferroptosis in NSCLC by directly targeting and inhibiting BCAT2, leading to mitochondrial dysfunction. This finding not only reveals BCAT2 as a novel target of verbascoside but also confirms its ability to induce apoptosis and ferroptosis in NSCLC cells.

Although CD20-directed B cell depletion has long been used in neuromyelitis optica spectrum disorder (NMOSD), high-quality, large-scale, randomized controlled trials remain limited. In this multicenter, randomized, double-blind phase 3 trial, we evaluated obinutuzumab β (MIL62), a novel glycoengineered type II anti-CD20 monoclonal antibody, in patients with NMOSD. Eligible participants aged 18-70 years with aquaporin-4-immunoglobulin G (AQP4-IgG)-seropositive NMOSD and an Expanded Disability Status Scale (EDSS) score of 7.0 or lower were randomly assigned (1:1) to receive intravenous obinutuzumab β 1,000 mg (n = 45) or placebo (n = 46). The primary outcome-time to first adjudicated relapse on or before week 52-was met: relapse occurred in two of 45 (4.4%) obinutuzumab β-treated participants and in 21 of 46 (45.7%) placebo-treated participants (hazard ratio = 0.069, 95% confidence interval: 0.016-0.296, P < 0.0001). The incidence of grade 3 or higher treatment-related adverse events was similar between the obinutuzumab β group (6.7%) and the placebo group (6.5%). These findings support obinutuzumab β as a glycoengineered type II anti-CD20 therapeutic option for patients with AQP4-IgG+ NMOSD. ClinicalTrials.gov identifier: NCT05314010 .

OBJECTIVE:Neuromyelitis optica (NMO) is a severe autoimmune disorder of the central nervous system (CNS) characterized by aquaporin-4 antibody (AQP4-IgG)-mediated astrocyte injury. IL-1β-mediated inflammatory signaling plays a critical role in amplifying astrocyte damage and propagating CNS inflammation in NMO. However, the astrocyte-intrinsic mechanisms linking IL-1β signaling to downstream pathways, such as STING activation, remain poorly understood. To address this knowledge gap, in this study, we aim to elucidate the astrocyte-intrinsic mechanisms, specifically the IL-1β-IL-1R STING signaling axis, that contribute to NMO pathogenesis, and to evaluate the therapeutic potential of IL-1β-targeting antisense oligonucleotides (ASOs). METHODS:Using a multi-level experimental system comprising in vitro primary astrocytes, ex vivo organotypic cerebellar slices, and in vivo NMO mouse models, we systematically investigated the critical role of the astrocytic IL-1β-IL-1R STING signaling axis in NMO pathogenesis. Utilizing diverse interventions-including an IL-1β-neutralizing antibody, astrocyte-specific IL-1β knockout, the IL-1R inhibitor Anakinra, STING genetic ablation, and IL-1β ASOs-in conjunction with behavioral, histopathological, and molecular analyses, we comprehensively delineated the impact of this signaling pathway on NMO pathology. These data support the translation of targeted therapeutic strategies. RESULTS:IL-1β signals through the IL-1 receptor (IL-1R) to induce STING-dependent proinflammatory cytokine production in astrocytes. This inflammatory cascade can be suppressed by the IL-1R antagonist anakinra or genetic ablation of STING. Therapeutic administration of lead IL-1β targeting ASO reduces IL-1β expression, preserves AQP4 levels and myelin integrity, and improves functional outcomes. INTERPRETATION:The IL-1β-IL-1R STING signaling axis is a central contributor to NMO pathogenesis and supports IL-1β ASO therapy as a promising potential disease-modifying approach. ANN NEUROL 2026;100:487-510.

To evaluate the feasibility and safety of a renal vein Doppler-guided central venous pressure (CVP) tritration protocol (RVD-CVP) in patients with sepsis, and to explore preliminary kidney-related and hemodynamic signals. This prospective, single-center, parallel-group pilot randomized controlled trial was conducted in an ICU between April and December 2023. Adults with sepsis and CVP monitoring were randomized 1:1 to RVD-CVP (renal vein Doppler assessments at 0, 6, 24, 48, and 72 h with prespecified CVP targets to maintain, lower, or defer) or usual care without renal vein Doppler guidance. The primary outcome was feasibility, defined as complete protocol delivery in the intervention group. Secondary outcomes were exploratory. Of 173 screened patients, 127 were randomized (63 intervention; 64 control). Complete protocol delivery was achieved in 63/63 intervention patients (100.0

OBJECTIVE:This study aimed to investigate the protective effect and mechanism of Astragalus polysaccharide (APS) on autoimmune encephalomyelitis. METHODS:C57BL/6 mice were randomly divided into the blank control group, EAE group, and APS intervention group (n=15/group). The Experimental Autoimmune Encephalomyelitis (EAE) mouse model was established by active immunization. The pathological changes in the spinal cord were evaluated by Hematoxylin-eosin (HE) and Luxol Fast Blue (LFB) staining. The number of CD11b+ Gr-1+ myeloid-derived suppressor cells (MDSCs) in the spleen tissues of mice in each group was determined by immunofluorescence staining. The expression of Arginase-1 in the spinal cord and spleen of each group was detected by immunofluorescence double staining. The TNF-α, IL-6, and Arginase-1 levels in the spleen were detected by ELISA assay. A western blot was used to detect the protein expression of the AMPK/JAK/STAT3/Arginase-1 signaling pathway. RESULTS:After the intervention of APS, the incidence of autoimmune encephalomyelitis in mice of the APS group was significantly lower than that in the EAE group, and the intervention of APS could significantly delay the onset time in the EAE mice, and the score of neurological function deficit in mice was significantly lower than that in EAE group (P < 0.05). APS intervention could reduce myelin loss and improve the inflammatory response of EAE mice. Moreover, it could induce the expression of CD11b+ GR-1 + bone MDSCs in the spleen and increase the expression of Arginase-1 in the spinal cord and spleen. This study further demonstrated that APS can protect EAE mice by activating the AMPK/JAK/STAT3/Arginase-1 signaling pathway. CONCLUSION:After the intervention of APS, myelin loss and inflammatory response of EAE mice were effectively controlled. APS promoted the secretion of Arginase-1 by activating MDSCs and inhibited CD4+T cells by activating AMPK/JAK/STAT3/Arginase-1 signaling pathway, thus improving the clinical symptoms and disease progression of EAE mice.

Aims: The aim of this study was to systematically analyze the role of Brucea javanica in the treatment of cervical cancer (CC) and its underlying mechanisms by means of network pharmacology and molecular docking. Background: Brucea javanica is a traditional Chinese herbal medicine used for the treatment of malaria and cancers, but its mechanism of action in CC is unknown. Objective: The objective of the study is screening of active chemical constituents of Brucea javanica by Traditional Chinese Medicine Systems Pharmacology (TCMSP) database and investigating their potential targets involved in CC therapy. Methods: The GeneCards database was used for the disease targets of CC, the drug-compound-disease target network was constructed by using the Cytoscape 3.8.0 software. Then, the key targets in the protein-protein interaction (PPI) network were identified, and the "clusterProfiler" was used for the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The qRT-PCR, CCK-8, and flow cytometry were used to assess the expression levels of specific target genes in CC cells, as well as their effects on cell proliferation, apoptosis, and reactive oxygen species (ROS) levels, respectively. Protein-compound complex analysis was performed using molecular dynamics simulation. Results: A total of 15 active compounds and their 86 treatment targets were obtained from the Brucea javanica analysis, in which 51 target genes were associated with the CC-related disease targets. Then, a PPI analysis identified 12 key genes (including EGFR, TP53, BCL2, AKT1, JUN, TNF, CASP3, IL6, MMP9, ERBB2, CCND1, and PTGS2) that were related to oxidative stress, PI3K-Akt, IL-17, p53, and JAK-STAT pathways, inflammatory response, and apoptosis pathways. In addition, AKT1 showed upregulation at the mRNA level in SiHa cells, and the knockdown of AKT1 significantly reduced the proliferation of CC cells and increased apoptosis and ROS levels. Molecular docking and dynamics simulations revealed a close binding between the active compounds and targets. Conclusions: The present research comprehensively examined the active compounds, potential targets, and pathways of Brucea javanica in CC treatment, providing a novel insight for CC treatment.

IntroductionPeripheral blood immune cell profiles of neuromyelitis optica spectrum disorder (NMOSD) are still unclear under different disease states and after B cell depletion therapy. Moreover, NMOSD is often confused with multiple sclerosis (MS) and myelin oligodendrocyte glycoprotein antibody disease (MOGAD). The study aims to illustrate peripheral blood immune cell profiles of NMOSD under different disease states, after B cell depletion therapy, and compared with MS and MOGAD.MethodsThis study included 76 NMOSD patients, 20 MS patients, and 12 MOGAD patients in the acute phase, along with 37 controls whose sex and age were matched with NMOSD patients. Forty-two of 76 patients with acute NMOSD were followed in remission, of whom 13, 15, and 11 patients received rituximab treatment, inebilizumab treatment, or conventional immunosuppressive therapies alone, respectively. The levels of diverse peripheral blood immune cells were measured by blood routine examination and flow cytometry. Distinctions among groups were analyzed using statistical methods.ResultsCompared with controls and NMOSD patients in remission, there was an elevation in the levels of neutrophils, platelets, platelet/lymphocyte ratio, neutrophil/lymphocyte ratio, and systemic inflammation index in acute NMOSD patients, while a decline was observed in the levels of lymphocytes, eosinophils, basophils, CD3+ T cells, CD3+CD4+ T cells, and CD4/CD8 ratio. NMOSD had increased levels of platelets and platelet/lymphocyte ratio, and decreased levels of eosinophils, basophils, CD4/CD8 ratio, and CD3+CD4+ T cells compared with MS. NMOSD had decreased levels of eosinophils, basophils, and CD19+ B cells, along with elevated platelet/lymphocyte ratio compared with MOGAD. After rituximab treatment, not only did CD19+ B cell level decrease, but eosinophil counts also increased. After inebilizumab treatment, not only did CD19+ B cell level decrease, but also the ratios of CD3+ T cells and CD3+CD8+ T cells increased. The quantity and ratios of eosinophils in rituximab group surpassed those in inebilizumab group.DiscussionPeripheral blood immune cell profiles of acute NMOSD patients showed widespread distinctions compared to those of remission NMOSD patients, acute MS patients, acute MOGAD patients, and controls, as well as after differential therapies. Our findings provide clues to comprehensively understand the abnormality of the dynamic and integrated immune network in NMOSD, distinguish NMOSD from MS and MOGAD, and search for more effective and safe therapeutic targets.

ObjectivesThis case report aims to highlight the atypical presentation and management of a patient diagnosed with CASPR2-associated autoimmune encephalitis and NF155 antibody-positive autoimmune nodopathies (AN), initially presenting with limb weakness and hyponatremia.MethodsThe patient was identified through clinical evaluation and diagnostic testing including serum and cerebrospinal fluid analysis, neuroimaging, and nerve conduction studies.ResultsThe patient exhibited limb weakness, hyponatremia, cognitive abnormalities, and peripheral nerve involvement. Diagnostic testing revealed CASPR2 and NF155 antibody are positive. Therapeutic interventions included corticosteroids, plasma exchange, and intravenous immunoglobulin therapy, followed by B-cell depletion therapy. Treatment led to improvement in walking function and normalization of antibodies.DiscussionThis case report contributes to the literature by documenting a rare co-occurrence of CASPR2-associated autoimmune encephalitis and NF155 antibody-positive AN, with a unique presentation of hyponatremia. The findings underscore the importance of considering autoimmune etiologies in patients presenting with hyponatremia and neurological symptoms. Moreover, the favorable response to B-cell depletion therapy suggests a potential treatment option for similar cases. The main take away is the need for heightened clinical suspicion and comprehensive diagnostic evaluation in patients with complex neurological presentations, to facilitate timely diagnosis and appropriate management.

The high glucose-induced endothelial-mesenchymal transition (EndMT) may be the initial and underlying mechanism of diabetic vascular complications. Although tauroursodeoxycholic acid (TUDCA) plays various protective roles in diabetes and its complications, it's unclear whether TUDCA inhibits the high glucose-induced EndMT. In this study, human umbilical vein endothelial cells (HUVECs) were treated with high glucose and intervened with TUDCA. The mRNA expression of fibroblast as well as endothelial markers, fibroblast specific protein 1 (FSP1), collagen I, CD31 and calcium adhesion protein 5, was detected. The protein content of FSP1 and CD31 was ascertained, with FSP1 distribution illustrated. The scratch assay was performed to evaluate the migratory ability of HUVECs. The protein content of TGF-β1 and Smad3, the distribution of Smad3 and the binding of Smad3 to the gene promoter of FSP1, were measured. The results firstly showed that TUDCA reversed the expression of EndMT-related genes in high glucose-treated HUVECs. Furthermore, TUDCA reduced FSP1 content with elevation in CD31, inhibited FSP1 distribution and attenuated morphological changes of high glucose-treated HUVECs. Meanwhile, TUDCA inhibited the high glucose-enhanced migratory ability of HUVECs. Mechanically, TUDCA prevented the binding of Smad3 to the gene promoter of FSP1 in high glucose-treated HUVECs, although it had little effect on the content of TGF-β1 and Smad3. In conclusion, TUDCA inhibited the high glucose-induced EndMT via preventing Smad3 from binding to the gene promoter of fibroblast markers, such as FSP1, in HUVECs.

This study explores the effect of total secondary ginsenosides(TSG)on apoptosis and energy metabolism in H9c2 cells under hypoxia and its potential mechanisms.H9c2 cell viability was observed and the apoptosis rate was calculated to determine suitable intervention concentrations of TSG,antimycin A complex(AMA),and coenzyme Q10(CoQ10),along with the duration of hypoxia.H9c2 cells at the logarithmic phase were divided into a normal group,a model group,a TSG group,an AMA group,a TSG+AMA group,and a CoQ10 group.All groups,except the normal group,were treated with their respective intervention drugs and cultured under hypoxic conditions.Adenosine triphosphate(ATP)content and creatine kinase(CK)activity were measured using an ATP chemiluminescence assay kit and a CK colorimetric assay kit.Flow cytometry was used to assess apoptosis rates,and Western blot evaluated the expression levels of apoptosis-related proteins,including B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),cysteinyl aspartate-specific protease(caspase)-3,caspase-8,and caspase-9,as well as mitochondrial biogenesis-related proteins peroxisome proliferator-activated receptor-γ coactivator 1α(PGC-1α),estrogen-related receptor-α(ERRα),nuclear respiratory factor(NRF)-1,NRF-2,peroxisome proliferator activated receptor-α(PPARα),and Na+-K+-ATPase.RT-PCR was employed to analyze the mRNA expression of mitochondrial biogenesis factors,including PGC-1α,ERRα,NRF-1,NRF-2,PPARα,mitochondrial transcription factor A(TFAM),mitochondrial cytochrome C oxidase 1(COX1),and mitochondrial NADH dehydrogenase subunit 1(ND1),ND2.The selected intervention concentrations were 7.5 μg·mL-1 for TSG,10 μmol·L-1 for AMA,and 1×10-4 mol·L-1 for CoQ10,with a hypoxia duration of 6 h.Compared with the normal group,the model group showed decreased ATP content and CK activity,increased apoptosis rates,decreased Bcl-2 expression,and increased Bax,caspase-3,caspase-8,and caspase-9 expression in H9c2 cells.Additionally,the protein and mRNA expression levels of mitochondrial biogenesis-related factors(PGC-1α,ERRα,NRF-1,NRF-2,PPARα),mRNA expression of TFAM,COX1,and ND1,ND2,and protein expression of Na+-K+-ATPase in mitochondrial DNA,were also reduced.In the TSG and CoQ10 groups,ATP content and CK activity increased,and apoptosis rates decreased compared with those in the model group.The TSG group showed decreased protein expression of apoptosis-related proteins Bax,caspase-3,caspase-8,and caspase-9,increased protein and mRNA expression of mitochondrial biogenesis factors PGC-1α,ERRα,NRF-1,and PPARα,and increased NRF-2 protein expression and TFAM mRNA expression in mitochondrial DNA.Conversely,in the AMA group,ATP content and CK activity decreased,the apoptosis rate increased,Bcl-2 expression decreased,and Bax,caspase-3,caspase-8,and caspase-9 expression increased,alongside reductions in PGC-1α,ERRα,NRF-1,NRF-2,PPARα protein and mRNA expression,as well as TFAM,COX1,ND1,ND2 mRNA expression and Na+-K+-ATPase protein expression.Compared with the TSG group,the TSG+AMA group exhibited decreased ATP content and CK activity,increased apoptosis rates,decreased Bcl-2 expression,and increased Bax,caspase-3,caspase-8,and caspase-9 expression,along with decreased PGC-1α,ERRα,NRF-1,NRF-2,and PPARα protein and mRNA expression and TFAM,COX1,and ND1,ND2 mRNA expression.Compared with the AMA group,the TSG+AMA group showed increased CK activity,decreased apoptosis rate,increased Bcl-2 expression,and decreased Bax,caspase-8,and caspase-9 expression.Additionally,the protein and mRNA expression of PGC-1α,ERRα,NRF-1,PPARα,mRNA expression of TFAM,COX1,ND1,ND2,and Na+-K+-ATPase protein expression increased.In conclusion,TSG enhance ATP content and CK activity and inhibit apoptosis in H9c2 cells under hypoxia,and the mechanisms may be related to the regulation of PGC-1α,ERRα,NRF-1,NRF-2,PPARα,and TFAM expression,thus promoting mitochondrial biogenesis.

This study investigated the role of Talin1 in regulating dendritic cell (DC) activation and the neuroprotective benefits of Talin1-knockdown DCs in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Bone marrow-derived DCs (BMDCs) were transduced with shTalin1 lentiviral vectors in vitro. Their morphological and biochemical profiles, surface molecules expression, cytokines production, capacity to induce T cell responses, as well as regulatory mechanisms, were comprehensively assessed. In vivo, Talin1-knockdown BMDCs loaded with the MOG35-55 peptide were administered preclinically and therapeutically to EAE mice, with subsequent evaluation of EAE development, inflammatory infiltration, demyelination, and Th/Treg responses. Results demonstrated that Talin1 knockdown significantly inhibited the activation of BMDCs, as evidenced by decreased expression of surface molecules (MHCII, CD80, CD86) and pro-inflammatory cytokines (IL-1β, IL-6, TNF-α), increased expression of anti-inflammatory cytokine (IL-10), differential morphology, ultrastructure, and biochemical characteristics, accompanied by limited ability to stimulate CD4+T cell proliferation and polarization toward Th1 and Th17 subsets. Moreover, RNA-sequencing revealed downregulation of immune/inflammation-related processes and pathways in Talin1-knockdown BMDCs. Mechanistically, inhibition of the TLR4/MyD88/NF-κB pathway in BMDCs contributed to these effects. In vivo, Talin1-knockdown BMDCs significantly delayed the pathogenesis & progression of EAE, alleviated their neurological deficits and pathology, decreased Th1 and Th17 lineage levels, and boosted the abundance of Treg cells. Collectively, these findings indicate that Talin1 orchestrates BMDCs activation, and Talin1-knockdown BMDCs protect against EAE by rebalancing Th1/Th17/Treg dynamics, suggesting a potential approach for the development of precision therapies for MS and other autoimmune disorders.

ABSTRACT Objective To investigate whether nicotinamide adenine dinucleotide (NAD+) supplementation can improve behavioral changes in a cuprizone‐intoxicated mouse model. Methods Six‐week‐old C57BL/6J mice were divided into three groups: two were fed 0.2% cuprizone chow (cuprizone and cuprizone + NAD+ groups), and the other group was fed normal rodent chow (control group) for 4 weeks. The mice in the cuprizone + NAD+ group received 250 mg/kg/day NAD+ intraperitoneally once a day, while the other mice were administered saline simultaneously. Behavioral tests for spatial memory (Morris water maze and Y maze), locomotor ability (grip test and rotarod test), depression‐like behavior (open field test and tail suspension test), and exploratory behavior (open field test) were conducted. Results In the probe test of the Morris water maze, the cuprizone group spent a significantly smaller proportion of time in the target quadrant than the control group did (16.32% vs. 31.66%, p = 0.006). However, supplementation with NAD+ increased the value (28.78% vs. 16.32%, p = 0.023). Similarly, in the Y maze test, the cuprizone group demonstrated a notably lower ratio of effective alterations compared to the control group (0.543 vs. 0.648, p < 0.001), and the cuprizone + NAD+ group presented an improved ratio compared with the cuprizone group (0.613 vs. 0.543, p = 0.021). Compared with the control group, cuprizone toxicity resulted in a decreased time to fall (169.10 vs. 247.60 s, p = 0.015) in the grip test, but NAD+ supplementation mitigated this effect (261.60 vs. 169.10 s, p = 0.003). There were no significant differences in the immobile time among groups in both the tail suspension test and the open field test, and there were also no significant differences in center distance in the open field test. Conclusions Direct NAD+ supplementation improves the locomotor ability and spatial memory of cuprizone‐intoxicated C57BL/6J mice. However, NAD+ supplementation does not show significant effects on depressive and exploratory behavior of experimental mice.

A significant proportion of children with Autism Spectrum Disorder (ASD) present with immune imbalances. In this study, we sought to alleviate the core symptoms of autism by addressing immune dysregulation, especially T-cell subpopulation imbalances, in BTBR mice through low-dose IL-2 (LdIL-2) treatment. LdIL-2 (30,000 IU) was administered subcutaneously, and changes in autistic behaviors were observed before and after treatment. Behavioral assessments included the three-chamber test, self-grooming test, sniffing test, marble burying test, open field test and Y-maze test. We also examined alterations in peripheral Th/Treg ratios, cytokine levels, and M1/M2 microglia ratios in the central nervous system via flow cytometry. Neuroinflammatory proteins in cerebrospinal fluid were assessed using proteomic analysis. Furthermore, CD25+ Treg cells were depleted using PC61, followed by LdIL-2 intervention, to determine the role of Treg cells in LdIL-2-treated BTBR mice. Our results demonstrated that LdIL-2 significantly ameliorated core symptoms of autism in BTBR mice. LdIL-2 treatment increased Treg cell levels, restored Th17/Treg and Tfh/Treg balance, and corrected immune dysregulation. Central nervous system inflammation was reduced in mice. However, the behavioral improvements were diminished when Treg cells were depleted by PC61. This study represents the first attempt to treat ASD using LdIL-2. The treatment proved safe and effective in improving both core symptoms and immune imbalances in autism. Symptom improvement was linked to increased Treg cell levels in peripheral blood. LdIL-2 shows potential as a novel therapy for addressing core symptoms of autism.

Lung adenocarcinoma (LA) ranks among the most common malignant tumors worldwide. M2 macrophage-derived exosomes have been implicated in regulating LA progression and drug resistance. However, whether proteins encapsulated in these exosomes contribute to gefitinib resistance in LA remains to be elucidated. This study investigates the role of tissue inhibitor of metalloproteinase 1 (TIMP1) in mediating LA resistance to gefitinib. We demonstrated that M2 macrophages-derived exosomes enhanced the sensitivity of gefitinib-resistant LA cell lines (P < 0.05). Bioinformatics analyses validated the correlation of TIMP1 expression with LA progression and patient survival. Depletion of TIMP1 in M2 macrophage-derived exosomes suppressed the proliferation of gefitinib-resistant LA cells. Additionally, we confirmed that TIMP1 interacts with cluster of differentiation 74 (CD74), a process linked to the proliferation and migration of gefitinib-resistant LA cells. Additionally, this study validated that TIMP1 mediates the interaction between CD74 and macrophage migration inhibitory factor (MIF), potentially activating the PI3K/AKT signaling pathway. Collectively, these findings demonstrate that TIMP1 in M2 macrophage-derived exosomes facilitate the binding of MIF to CD74 in LA cells, thereby attenuating gefitinib resistance.

ABSTRACT Objective To investigate the therapeutic effects of liraglutide, a glucagon‐like peptide‐1 receptor agonist, on clinical progression and splenic T‐cell subsets in experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis. Methods EAE was induced in C57BL/6 mice via myelin oligodendrocyte glycoprotein immunization. The mice were divided into three groups: healthy controls (n = 8), EAE (n = 10), and EAE + liraglutide (n = 10). The EAE + liraglutide treatment group received subcutaneous liraglutide (10 µg/kg every other day) starting at 8 days postimmunization. Body weight and disease score were monitored for 30 days, and part of the animals (n = 4 for each group) were sacrificed 20 days postimmunization for flow cytometry of splenocytes, and splenic T helper 1 (Th1) and regulatory T (Treg) cell proportions were analyzed. Results Liraglutide significantly reduced maximum clinical scores (EAE 2.50 ± 0.41 versus EAE + liraglutide 1.75 ± 0.59, p = 0.005), but no significant differences in body weight were observed between the EAE and EAE + liraglutide groups. EAE induction decreased the proportion of splenic Treg cells (6.38% ± 0.72% versus control 10.55% ± 0.87%, p = 0.013), with liraglutide treatment showing no significant effect on Treg proportion relative to EAE alone (7.53% ± 1.54% versus 6.38% ± 0.72%, p = 0.975). However, the proportion of Th1 cells significantly increased after EAE induction (11.95% ± 1.58% versus control 6.05% ± 3.23%, p = 0.025), which was reduced by liraglutide (5.94% ± 2.53% versus EAE, p = 0.025). Conclusions The present preliminary findings demonstrate that liraglutide alleviates EAE severity, probably through peripheral immunomodulatory effects of splenic T helper cells.