Aging significantly impairs vaccine efficacy in older adults, driven by immunosenescence, inflammaging, and disruptions in the gut microbiota-mTOR-immune axis. This review synthesizes current evidence on how aging alters vaccine-induced immune responses through the interplay of gut microbiota dysbiosis and dysregulated mTOR signaling. Age-related microbial diversity declines and reduced short-chain fatty acid (SCFA) production exacerbate inflammation, while heightened mTOR activity suppresses autophagy, promotes pro-inflammatory states, and impairs T/B cell function, collectively diminishing antibody production and immune memory formation. We highlight the bidirectional interaction between SCFAs and mTOR, where SCFAs mitigate mTOR overactivation to enhance immune regulation, and mTOR dysregulation further aggravates microbial dysbiosis, forming a vicious cycle. Critically, this review systematically stratifies the evidence, distinguishing preclinical mechanistic insights from correlative human data. Animal and human studies suggest that targeting this axis-via mTOR inhibitors, probiotics, or dietary interventions-holds promise for improving vaccine responses in the elderly. We propose future research directions, including personalized vaccine strategies leveraging microbiota profiling and mTOR modulation, to address the challenges of infection in aging populations and advance precision medicine for healthy aging.

In the pathological cascade of cerebral ischemia, the pyroptosis axis mediated by the NLRP3 inflammasome in activated microglia is a core link driving neuroinflammation and secondary brain injury. Quercetin has been proven to possess multi-target neuroprotective activity, and its anti-inflammatory effect has attracted particular attention. However, direct molecular evidence is lacking regarding how quercetin precisely regulates the NLRP3/Caspase-1/GSDMD core pyroptosis axis in microglia in cerebral ischemia models and whether it can directly target NLRP3 to inhibit this axis, thereby alleviating cerebral ischemic injury. This study aimed to investigate the molecular mechanism by which quercetin alleviates cerebral ischemic injury through inhibiting the pyroptosis axis, combining cellular and animal models with molecular docking and molecular dynamics simulations. The oxygen-glucose deprivation (OGD) model of BV2 microglia and the photothrombotic (PT) model of focal cortical ischemia in male C57BL/6 mice were used to detect the ameliorative effect of quercetin on cerebral ischemia-related injury through cellular and animal experiments. AutoDock Vina 1.5.7 and GROMACS 2025.3 software were employed for molecular docking and molecular dynamics simulations, respectively, to analyze the binding mode and complex stability between quercetin and the NLRP3 protein. The results showed that quercetin could significantly ameliorate OGD-induced injury in BV2 cells and downregulate the expression of pyroptosis and inflammation-related proteins and factors. Meanwhile, it relieved motor dysfunction in PT mice, attenuated cortical neuronal injury, and inhibited the activation of the cerebral pyroptosis axis. At the molecular level, molecular simulation predictions indicated that quercetin might specifically bind to the NACHT domain of the NLRP3 protein, forming a complex with a stable conformation, and van der Waals interactions served as the main driving force for binding. This study confirmed that quercetin can directly bind to the NLRP3 protein and alleviate cerebral ischemia-induced inflammatory injury by inhibiting the activation of the NLRP3/Caspase-1/GSDMD pyroptosis axis and the release of downstream inflammatory factors. Combined with the molecular simulation results, a predictive hypothesis is proposed: direct binding of quercetin to the NLRP3 protein is one of its core mechanisms of action. These findings provide direct experimental evidence for the development of NLRP3-based drugs against ischemic brain injury.

Background: Mitophagy is a critical mitochondrial quality control mechanism that limits neuronal injury following cerebral ischemia/reperfusion injury (CI/RI). Tetramethylpyrazine (TMP), a bioactive alkaloid from Ligusticum chuanxiong Hort., exhibits neuroprotective effects in cerebrovascular disorders. However, whether these effects involve mitophagy regulation remains unclear. Methods: CI/RI was induced using a middle cerebral artery occlusion/reperfusion (MCAO/R) model in mice and an oxygen-glucose deprivation/reoxygenation (OGD/R) model in HT22 cells. Neurological function, infarct volume, mitochondrial function, and mitophagy-related markers were assessed. Pharmacological inhibitors and genetic manipulation of YAP and Parkin were used to investigate underlying mechanisms. Results: TMP treatment significantly reduced infarct volume and improved neurological deficits in MCAO/R mice, accompanied by enhanced mitophagy, as indicated by increased mitochondrial LC3 recruitment and Parkin expression. In OGD/R-injured HT22 cells, TMP promoted mitophagosome and mitolysosome formation, reduced mitochondrial reactive oxygen species, and restored mitochondrial membrane potential. Inhibition of mitophagy with Mdivi-1 attenuated TMP-mediated neuroprotection. Mechanistically, TMP promoted YAP nuclear localization, and inhibition of YAP or silencing of Parkin abolished TMP-induced mitophagy, while Parkin overexpression restored mitophagy under YAP inhibition. Conclusions: TMP alleviates CI/RI by promoting mitophagy through the YAP/Parkin signaling pathway, suggesting mitophagy modulation as a potential therapeutic strategy for ischemic brain injury.

Background/Objectives: Insomnia is a prevalent clinical sleep disorder, with existing hypnotic therapies limited by safety concerns. There is an urgent clinical need for new safe, effective sleep-promoting candidates derived from natural products. Spinosin is one of the main active components of Semen Ziziphi Spinosae that exerts sedative and hypnotic effects. The adenosine receptor (AR) has been reported as a potential therapeutic target for insomnia; however, the hypnotic effect of spinosin through the A2AR remains to be elucidated. Methods: In the study, the involvement of A2ARs in spinosin’s hypnotic effect was investigated using caffeine and further elucidated in A2AR-knockout (KO) mice. Diazepam was used as a positive control drug to validate the experimental model and evaluate the hypnotic effect of spinosin. Molecular docking and molecular dynamics (MDs) simulations were performed to validate the interaction of spinosin with the A2AR. Results: The hypnotic effects of spinosin were effectively antagonized by caffeine. Compared with A2AR-wild-type (WT) mice, spinosin-induced non-rapid eye movement (NREM) sleep and locomotor activity diminution were significantly reduced in A2AR-KO mice. Spinosin significantly increased the activity of γ-aminobutyric acid (GABA)ergic medium spiny neurons (MSNs) in the nucleus accumbens (NAc) and significantly decreased the activity of orexin neurons in the lateral hypothalamus (LH), as revealed by c-Fos immunostaining. These effects were significantly reversed by caffeine pretreatment or in A2AR-KO mice. Finally, the results of molecular docking showed that spinosin had a good binding potential with the A2AR. MD simulations further demonstrated that spinosin had strong binding stability with the A2AR. Conclusions: Our findings strongly suggest that spinosin exerts the hypnotic effects through the A2AR, and thus may have therapeutic potential for insomnia. Our identification of spinosin’s direct molecular target supports its translational potential as a novel natural-origin candidate for clinical insomnia drug development.

The neurovascular unit (NVU) is composed of a diverse array of cells and an extracellular matrix (ECM). Neural cells and blood vessels are intricately interconnected, forming a cohesive whole. Specific cellular components and structures within the NVU play an indispensable role in maintaining homeostasis of the central nervous system (CNS). With the advancement and maturation of cell co-culture technology, various three-dimensional (3D) NVU models continue to emerge, offering a more objective and comprehensive perspective for in vitro studies of CNS diseases. Specifically, these 3D NVU models include Transwell Chamber models, gel-polydimethylsiloxane (PDMS)-based 3D models, self-assembled NVU models and microfluidic NVU models, which reconstruct the complex NVU architecture to varying degrees. This review systematically summarizes multiple 3D construction strategies for in vitro NVU to overcome the limitations of conventional cellular tests or animal experiments, highlights the critical roles of biomimetic gel in recapitulating native cell-gel crosstalk, comparatively analyzes four major 3D NVU technical routes in terms of cellular composition, vascular morphology, barrier performance, and reproducibility, categorizes application scenarios of 3D NVU platforms oriented to practical research demands, including oxygen-glucose deprivation/reoxygenation (OGD/R) injury modeling, blood-brain barrier (BBB) permeability assay, CNS drug penetration screening, neuroinflammation and neurotoxicity evaluation, proposes practical principles for model selection under different experimental purposes, and concludes with current bottlenecks, including imperfect vascular network maturation and lack of unified evaluation criteria, together with future perspectives for standardized 3D NVU in vitro. By comparing the advantages and limitations of these approaches, we aim to clarify their optimal applicability for investigating specific pathological mechanisms and screening potential therapeutics.

Bone marrow mesenchymal stem cell-derived extracellular vesicles (BMSC-EVs) show therapeutic promise for ischemic stroke (IS). Preconditioning MSCs with drugs can modulate the cargo composition and function of their derived EVs. This study investigated the therapeutic effects and underlying mechanisms of EVs derived from tetramethylpyrazine (TMP)-preconditioned BMSCs (TMP-BMSC-EVs) in IS. EVs were isolated from BMSCs pretreated with or without TMP by differential centrifugation. The therapeutic efficacy of EVs was evaluated in a rat model of middle cerebral artery occlusion (MCAO) through neurological function assessments and infarct volume quantification. The expression of miR-486 and its roles in regulating microglia/macrophage polarization and neurogenesis, as well as the mechanistic targets, were examined by real-time quantitative polymerase chain reaction (RT-qPCR), immunofluorescence staining, and Western blotting. TMP-BMSC-EVs exerted superior therapeutic efficacy compared to BMSC-EVs. Mechanistically, TMP-BMSC-EVs were enriched with miR-486, which promoted microglia/macrophage M2 polarization and neurogenesis, while downregulating phosphatase and tensin homolog (PTEN) and phosphorylated NF-κB (p-NF-κB) protein levels, and upregulating phosphorylated Akt (p-Akt) expression. Transfection with a miR-486 inhibitor abolished the beneficial effects of TMP-BMSC-EVs, which could be counteracted by the PTEN inhibitor bisperoxovanadium (bpV). TMP-BMSC-EVs could significantly promote neural repair by driving microglia/macrophage M2 polarization and enhancing neurogenesis through miR-486-mediated PTEN inhibition, thereby offering a promising treatment strategy for IS. The mechanism of TMP-BMSC-EVs promote function recovery in IS

5-Lipoxygenase (5-LOX) is implicated the pathogenesis of depression, yet the underlying molecular mechanism remains unclear. In our study, we investigated the dynamic changes of 5-LOX expression after chronic unpredictable mild stress (CUMS) stimulation. The results demonstrated that 5-LOX expression was significantly upregulated in the prefrontal cortex (PFC) after CUMS stimulation for 2 weeks, 4 weeks and 8 weeks. CUMS stimulation activated 5-LOX on neurons, and induced neuronal loss in the PFC. Moreover, the expression levels of pyroptosis-related proteins showed a gradual increase with prolongation of CUMS stimulation. Notably, inhibiting 5-LOX with zileuton effectively alleviated depressive-like behaviors, increased the expression of monoamine neurotransmitters and exerted neuroprotective effects in CUMS-exposed mice. 5-LOX inhibition also markedly reduced the expression levels of pyroptosis-related proteins. We further explored the role of 5-LOX in NLRP3-mediated cell pyroptosis in vitro. 5-LOX knockdown significantly suppressed LPS/ATP-induced pyroptosis in PC12 cells. 5-LOX overexpression enhanced the pyroptosis induced by LPS/ATP, while NLRP3 inhibitors reversed the aggravation. These findings indicate that 5-LOX contributes to the pathogenesis of depression via promoting NLRP3-mediated cell pyroptosis. Thus, 5-LOX inhibition could be applied as a clinic intervention for the treatment of depression.

Background:The biological process of neurogenesis involves the production of new and completely functional neurons in two specific regions of the brain: the ventricular-subventricular zone (V-SVZ) and the subgranular zone (SGZ) of the hippocampal dentate gyrus by neural stem cells (NSCs). Interfering with this mechanism harms the brain and may result in neurological disorders. Cell-based therapies are becoming recognized as optimal possibilities for facilitating neurogenesis. To comprehend the many processes and mechanisms of neurogenesis and the role of mesenchymal stem cells (MSCs) as active contributors to pathologic events influencing neurogenesis. We utilized the Web of Science (core collection) as the data source. Methods:The search was performed in the Web of Science core collection database until April 30, 2024, with the terms "Mesenchymal stem cells" as the title and "neurogenesis" as the topic. Results:A total of 407 papers about mesenchymal stem cells in neurogenesis published from 2004 to 2024 were retrieved. Further, we performed a bibliometric analysis of these publications, such as generating cooperation maps, co-citation analysis of journals and references, and cluster analysis of keywords. Next, we discussed the mechanism by which MSCs promote neurogenesis during the onset of Alzheimer's disease (AD) and stroke diseases. Conclusion:Overall, three aspects primarily reflect the treatment of stroke with MSCs: neural circuit reconstruction, mitochondrial transfer, and extracellular vesicle transfer. The treatment of AD with MSCs is mainly reflected in the five aspects of inhibiting neuroinflammation, microglia changes, amyloid-β removal, functional recovery of autophagy, and blood-brain barrier (BBB) function recovery. Finally, we also made prospects for future research of MSCs.

Spinosin, a key flavonoids component found in Semen Zizhiphi spinosae, is known to enhance pentobarbital-induced sleep, which is primarily assessed with the loss-of-righting reflex (LORR). This research focused on investigating the impact of spinosin on sleep regulation in typical murine models. We used electroencephalogram (EEG) and electromyogram (EMG) recordings to evaluate the effects of spinosin (10, 20, 40 mg/kg, i.p.) on sleep–wake state. Immunohistochemical techniques were employed to investigate the c-Fos expression in various sleep–wake brain regions following the injection of spinosin. In the initial three-hour period following administration, spinosin administered at a dose of 40 mg/kg exhibited a notable augmentation in the duration of non-rapid eye movement (NREM) sleep, with a 2.04-fold increase (P < 0.0001), accompanied by a reduction in wakefulness by approximately 42.84

Ganoderma lucidum polysaccharides (GLPs) exert antiobesity effects that are linked to gut-microbiota modulation, yet the underlying mechanisms remain elusive. Here, we show that GLPs raise the fecal butyrate level, which in turn inhibits high-fat diet (HFD)-induced weight gain, fat accumulation, adipocyte hypertrophy, and elevated serum triglyceride by promoting adipose triglyceride lipase (ATGL) expression to activate lipolysis. GLPs and butyrate also strengthen the intestinal barrier, reflected by elevated tight junction proteins and goblet cells, leading to reduced serum lipopolysaccharide-binding protein (LBP) and attenuated white-adipose-tissue (WAT) inflammation. The intestinal barrier enhancement involves FABP4-PPARγ signaling. In addition, the butyrate-enhancement and antiobesity effects of GLPs are abolished after antibiotic-mediated microbiota depletion. Notably, GLPs selectively enrich the abundance of Lactobacillus, especially Lactobacillus johnsonii, whose supplementation alone increases fecal butyrate and recapitulates GLPs-induced benefits by reinforcing gut barrier integrity and lipolysis. Collectively, our findings identify the L. johnsonii-butyrate axis as a central target through which GLPs attenuate obesity and associated inflammation.

Neuronal loss following ischemic stroke (IS) is a major cause of long-term neurological dysfunction. Neural stem cell (NSC) transplantation holds great potential for neural tissue repair, but exogenous NSCs often over-differentiate into astrocytes, resulting in glial scar formation and poor neuronal maturation, which limits clinical application. Astragaloside IV (AS-IV), a plant-derived monomer, has been shown to regulate the proliferation, migration, and differentiation of endogenous NSCs. This study investigated whether AS-IV preconditioning could promote the neurogenesis of transplanted NSCs and improve functional recovery after IS. Primary NSCs were cultured and exposed to various concentrations of AS-IV, and 100 μM was selected for further use. A rat model of middle cerebral artery occlusion/reperfusion (MCAO/R) was established, and AS-IV-preconditioned or untreated EGFP-labeled NSCs were transplanted into the striatum. Brain injury was assessed histologically. Neurological recovery was evaluated using the modified Neurological Severity Score, grip strength, adhesive removal, and Morris water maze tests. Immunofluorescence staining was used to assess graft survival, differentiation fate, synapse formation, and glial scar area. In vitro experiments explored the involvement of the miR-199a-5p/Caveolin-1 signaling pathway. AS-IV preconditioning significantly improved the survival and neuronal differentiation of grafted NSCs, reduced astrocytic lineage commitment and glial scar formation, and enhanced synaptic reconstruction. These effects were associated with improved sensorimotor and cognitive function. In conclusion, AS-IV pretreatment of NSCs promoted neuronal lineage commitment, inhibited glial scarring, and facilitated functional recovery after ischemic stroke. Targeting the miR-199a-5p/Caveolin-1 axis may offer a novel strategy for optimizing NSC-based therapy in stroke.

BACKGROUND:Ischemic stroke (IS) remains a challenge in clinical treatment due to limited therapeutic options. While artemisinin (ART), an antimalarial drug, shields against acute IS via anti-inflammatory, antioxidant, and anti-apoptotic properties, the long-term benefits and specific underlying mechanisms have not been fully elucidated. Here, we investigate whether ART ameliorates IS injury and promotes neurogenesis by activating the peroxisome proliferator-activated receptor γ (PPARγ)-dependent M2 microglial polarization. METHODS:The experimental models included transient middle cerebral artery occlusion/reperfusion (MCAO/R) in rats and oxygen-glucose deprivation/reoxygenation (OGD/R) in primary microglial cultures to simulate IS. The therapeutic effects of ART were evaluated by neurological functions and infarct volume. PPARγ inhibitor T0070907 (T007) was intraperitoneally injected 24 h following MCAO/R at a dose of 2 mg/kg in vivo and a concentration of 10 μM for 30 min before OGD in vitro. We utilized real-time quantitative polymerase chain reaction (RT-qPCR) along with Western blot analyses to detect the microglia markers and PPARγ. The proliferation and differentiation of neural stem cells (NSCs) both in vivo and in vitro were assessed via immunofluorescence labeling. The neurogenic potential of ART-treated microglia was investigated by conditioned medium. The levels of brain-derived growth factor (BDNF) and insulin-like growth factor-1 (IGF-1) in microglia were measured by immunofluorescence staining and enzyme-linked immunosorbent assay (ELISA). RESULTS:ART treatment significantly alleviated short- and long-term neurological deficits and reduced cerebral infarct volume in rats with IS. Experiments conducted both in vivo and in vitro experiments illustrated that ART directed microglia away from the pro-inflammatory M1 state towards the anti-inflammatory M2 state, enhanced neurogenesis, and upregulated the expression of PPARγ, BDNF, and IGF-1. In addition, the conditioned medium from ART-exposed microglia stimulated the proliferation and neuronal differentiation of primary NSCs. However, these positive effects were effectively counteracted by the use of PPARγ inhibitor T0070907 (T007). CONCLUSION:Our findings demonstrate that ART ameliorates IS injury and promotes neurogenesis mainly through PPARγ-mediated microglia M2 polarization. Therefore, ART can be considered a potential therapeutic drug for IS.

In recent years, TLR4, as a key member of the TLRs family, has attracted increasing attention for its role in neuroimmune diseases. As a critical immune surveillance molecule, TLR4 is considered a promising target for the modulation of neuroimmune disorders. A growing body of research has confirmed that artemisinin and its derivatives, as natural immunomodulators, participate in the regulation of TLR signaling pathways-particularly showing a close relationship with TLR4 among the TLR family.Recent studies have revealed a novel dual regulatory effect of artemisinin and its derivatives on the TLR4 signaling pathway. These compounds can not only selectively modulate TLR4-mediated inflammatory signaling cascades, such as the NF-κB pathway, effectively suppressing the excessive activation of microglia, but also inhibit TLR4 dimerization and endocytosis, thereby blocking downstream signal transduction and alleviating neuroinflammatory responses. This novel finding provides a molecular basis for targeting TLR4 in the treatment of neuroimmune diseases such as multiple sclerosis (MS) and Alzheimer's disease (AD).Moreover, as naturally derived TLR4 modulators, artemisinin and its derivatives show great potential as new lead compounds for targeted therapies in neuroimmune diseases, offering new avenues and strategies for treatment. Therefore, this review focuses on the molecular structural characteristics of artemisinin and its derivatives, and the TLR4-mediated signaling pathways. It explores their potential mechanisms of action in modulating neuroimmune disorders from multiple perspectives-including drug structural properties, molecular docking simulations, and target regulation. The objective is to identify safe and effective neuroimmune modulators from natural products and provide new research perspectives and strategies for the TLR4-targeted treatment of neuroimmune diseases.

There is a significant correlation between regulatory functions of orexin system and depressive symptoms. Parvalbumin (PV) interneurons, the main subtype of inhibitory g-aminobutyric acid (GABA) interneurons, are involved in the regulation of depression-like behaviors too. To provide several lines of evidence regarding neural connection between two types of inneurons the depression associated nuclei, a double-label immunohistochemical staining technique was employed to reveal overlap betweennd PV a orexin immunoreactivity in the mouse brain. Medial prefrontal cortex (mPFC) mainly consists of the cingulate cortex, area1), 1 (Cg prelimbic cortex (PrL) and infralimbic cortex (IL), in which regions contained similar obvious overlap between orexin d fibers PV an somas with axons. In the basal forebrain (BF), overlap was observed in the lateral septal nucleus, ventral part (LSV), theeus nucl accumbens shell (AcbSh), bed nucleus of the stria terminalis, ventral division (BSTV), ventral pallidum (VP), central d amygdaloi nucleus (Ce). In the diencephalon, besides the lateral hypothalamic area (LH) and dorsomedial hypothalamic nucleus (DM), chin whi two regions contained overlap between orexin positive perikarya with fibers and PV immunoreactivity, different densitiesxin of andore PV fibers were closely intermingled in the paraventricular hypothalamic nucleus (PVN), paraventricular thalamic nucleus arcuate(PVT), hypothalamic nucleus (Arc). In the brainstem, densities of intermixed orexin and PV fibers were from high to low in theraphe dorsal nucleus (MnR), and orexin buttons were embedded within PV fibers, as for the ventral tegmental area (VTA), overlap was observed between PV somas with axons and orexin fibers. Hence, PV and orexin neurons and its projections jointly target several braingions re involved in regulating depression, which provides anatomic evidence supporting coordinated PV/orexin actions within these brain regions.

Stroke is a prominent contributor to mortality and impairment on a global scale. Ischemic stroke accounts for approximately 80% of stroke cases and is caused by occlusion of cerebral blood vessels. Enhancing neurogenesis through the modulation of the neural stem cell niche in the adult brain is a promising therapeutic strategy for individuals afflicted with ischemic stroke. Neurogenesis results in the generation of newborn neurons that serve as replacements for deceased neural cells within the ischemic core, thereby playing a significant role in the process of neural restoration subsequent to cerebral ischemia. Research has shown that activation of the Wnt/β-catenin pathway can augment neurogenesis following cerebral ischemia, suggesting that this pathway is a potentially beneficial therapeutic target for managing ischemic stroke. This review provides an extensive analysis of the current knowledge regarding the involvement of the Wnt/β-catenin pathway in promoting neurogenesis, thereby offering a promising avenue for therapeutic intervention in the context of ischemic stroke or other neurological impairments.

Bone marrow mesenchymal stem cell-derived exosomes (BMSC-Exos) have been shown to promote angiogenesis after ischemic stroke, in which microRNAs (miRs) are believed to play an important role in exosome-mediated therapeutic effects, though the mechanism is still not clear. In this study, a series of molecular biological and cellular assays, both in vitro and in vivo, were performed to elucidate the role of exosomal miR-486 in angiogenesis following cerebral ischemic and its molecular mechanisms. Our results revealed that BMSC-Exos significantly improved neurological function and increased microvessel density in ischemic stroke rats. In vitro assays showed that BMSC-Exos promoted the proliferation, migration, and tube formation ability of oxygen-glucose deprivation/reoxygenation (OGD/R) injured rat brain microvascular endothelial cells (RBMECs). Importantly, BMSC-Exos increased the expression of miR-486 and phosphorylated protein kinase B (p-Akt) and down-regulated the protein level of phosphatase and tensin homolog (PTEN) in vivo and in vitro. Mechanistic studies demonstrated that transfection with miR-486 mimic enhanced RBMECs angiogenesis and increased p-Akt expression, while inhibited PTEN expression. On the other hand, the miR-486 inhibitor induced an opposite effect, which could be blocked by PTEN siRNA. It was thus concluded that exosomal miR-486 from BMSCs may enhance the functional recovery by promoting angiogenesis following cerebral ischemic injury, which might be related to its regulation of the PTEN/Akt pathway.

BACKGROUND:Brain tumors are one of the leading causes of epilepsy, and brain tumor-related epilepsy (BTRE) is recognized as the major cause of intractable epilepsy, resulting in huge treatment cost and burden to patients, their families, and society. Although optimal treatment regimens are available, the majority of patients with BTRE show poor resolution of symptoms. BTRE has a very complex and multifactorial etiology, which includes several influencing factors such as genetic and molecular biomarkers. Advances in multi-omics technologies have enabled to elucidate the pathophysiological mechanisms and related biomarkers of BTRE. Here, we reviewed multi-omics technology-based research studies on BTRE published in the last few decades and discussed the present status, development, opportunities, challenges, and prospects in treating BTRE.METHODS:First, we provided a general review of epilepsy, BTRE, and multi-omics techniques. Next, we described the specific multi-omics (including genomics, transcriptomics, epigenomics, proteomics, and metabolomics) techniques and related molecular biomarkers for BTRE. We then presented the associated pathogenetic mechanisms of BTRE. Finally, we discussed the development and application of novel omics techniques for diagnosing and treating BTRE.RESULTS:Genomics studies have shown that the BRAF gene plays a role in BTRE development. Furthermore, the BRAF V600E variant was found to induce epileptogenesis in the neuronal cell lineage and tumorigenesis in the glial cell lineage. Several genomics studies have linked IDH variants with glioma-related epilepsy, and the overproduction of D2HG is considered to play a role in neuronal excitation that leads to seizure occurrence. The high expression level of Forkhead Box O4 (FOXO4) was associated with a reduced risk of epilepsy occurrence. In transcriptomics studies, VLGR1 was noted as a biomarker of epileptic onset in patients. Several miRNAs such as miR-128 and miRNA-196b participate in BTRE development. miR-128 might be negatively associated with the possibility of tumor-related epilepsy development. The lncRNA UBE2R2-AS1 inhibits the growth and invasion of glioma cells and promotes apoptosis. Quantitative proteomics has been used to determine dynamic changes of protein acetylation in epileptic and non-epileptic gliomas. In another proteomics study, a high expression of AQP-4 was detected in the brain of GBM patients with seizures. By using quantitative RT-PCR and immunohistochemistry assay, a study revealed that patients with astrocytomas and oligoastrocytomas showed high BCL2A1 expression and poor seizure control. By performing immunohistochemistry, several studies have reported the relationship between D2HG overproduction and seizure occurrence. Ki-67 overexpression in WHO grade II gliomas was found to be associated with poor postoperative seizure control. According to metabolomics research, the PI3K/AKT/mTOR pathway is associated with the development of glioma-related epileptogenesis. Another metabolomics study found that SV2A, P-gb, and CAD65/67 have the potential to function as biomarkers for BTRE.CONCLUSIONS:Based on the synthesized information, this review provided new research perspectives and insights into the early diagnosis, etiological factors, and personalized treatment of BTRE.

Abstract Background Conventional observational designs face challenges in studying this relationship, as confounding factors, reverse causality, minor exposure factors and multiple tests cannot be completely eliminated. There is currently a lack of MR studies concerning immune cells and the risk of ischemic stroke. This particular study offers a novel perspective on risk prediction for ischemic stroke. Objective To investigate the causal relationship between immune cells and ischemic stroke through Mendelian randomization analysis. Methods A complete two-sample Mendelian randomization (MR) analysis was utilized to ascertain the causative relationship between immune cells and ischemic stroke. Using publicly available genetic data, we investigated the causal association between 731 immune cells and the risk of ischemic stroke. Four immune characteristics were included: relative cells (RC), absolute cells (AC), median fluorescence intensity (MFI), and morphological parameters (MP). MR-Egger, Weighted median, Inverse variance weighted (IVW), Weighted mode, Simple mode, and MRPRESS were utilized for analysis. Heterogeneity and horizontal pleiotropy tests were also conducted. Results Mendelian randomization analysis showed that 32 of the 731 immune cells had a robust causal relationship with ischemic stroke, among which 15 immune cells such as IgD−CD27− %B cell (β = 0.033, 95%CI = 1.002 ~ 1.065, p = 0.037), IgD+ CD24 + AC (β = 0.045, 1.010 ~ 1.082, p = 0.012), CD25hi CD45RA−CD4 not Treg %T cell (β = 0.022, 95%CI = 1.002 ~ 1.042, p = 0.028) and soon. CD62L−HLADR++ monocyte AC (β =-0.053, 95% CI = 0.914 ~ 0.985, p = 0.005), CD33br HLA DR+ CD14− AC (β =-0.017, 95% CI = 0.972 ~ 0.995, p = 0.004), EM DN (CD4−CD8−) %DN (β =-0.014, 95% CI = 0.975 ~ 0.997, p = 0.014), etc. There exists a strong inverse causal link for ischemic stroke. Conclusion Our study has demonstrated a close genetic link between immune cells and ischemic stroke. Fifteen immune cells such as IgD−CD27− %B cell, IgD+ CD24+ AC, CD25hi CD45RA−CD4 not Treg %T cell have robust positive causal associations with ischemic stroke, and seventeen immune cells such asCD62L− HLA DR++ monocyte AC, CD33br HLA DR+ CD14− AC, EM DN (CD4−CD8−) %DN have robust positive causal associations with ischemic stroke. A strong inverse causal relationship with ischemic stroke offers direction for forthcoming clinical studies.

Background In the pathological process of cerebral ischemia, the neuroinflammation triggered by NLRP3 inflammasome-mediated neuronal pyroptosis is considered a crucial factor contributing to brain damage. Although, previous research has revealed the anti-inflammatory and neuroprotective effects of quercetin (Que), its precise mechanisms in intervening and alleviating neuroinflammation and the pyroptosis axis triggered by cerebral ischemia remain to be fully elucidated. Objective To investigate the role and mechanism of Que in alleviating cerebral ischemia injury by modulating the NLRP3 inflammasome-mediated pyroptosis axis. Methods The oxygen-glucose deprivation and reperfusion (OGD/R) model with BV2 microglial cells and the photothrombosis (PT) model with C57/BL male mice were utilized. The study explored the neuroprotective effect of Que against cerebral ischemia injury through the regulation of the pyroptosis axis via the NLRP3 inflammasome, assessed through experiments including reactive oxygen species measurement, cell pyroptosis rate detection, behavioral tests, HE staining, Nissl staining, and examination of pyroptosis-related proteins and inflammatory factors. Results Que alleviated cell damage and pyroptosis induced by OGD/R, reduced intracellular reactive oxygen species levels, improved behavioral impairment in mice after modeling, decreased brain injury area, maintained neuronal morphology, and lowered the expression of pyroptosis axis-related proteins such as NLRP3, Caspase-1, ASC, GSDMD, as well as inflammatory factors IL-18, IL-1β, TNF-α. Conclusion Que alleviated cerebral ischemia injury by regulating the pyroptosis axis through inhibiting the activation of NLRP3 inflammasome. This provides robust experimental evidence for the anti-neuroinflammatory effects of Que and the development of neuroprotective drugs.