Post-stroke cognitive impairment (PSCI) is a frequent and disabling consequence of stroke that limits long-term recovery and quality of life. Disruption of the hippocampal neurogenic niche may impair adult hippocampal neurogenesis (AHN), while exercise is increasingly recognized as a promising non-pharmacological strategy for cognitive rehabilitation. This review synthesizes evidence that exercise may shift the post-stroke hippocampal niche from a hostile to a permissive state through coordinated regulation of neuroinflammation, neurovascular integrity, and mitochondrial homeostasis. We distinguish increased progenitor proliferation from productive neurogenesis, defined by the survival, maturation, appropriate positioning, and functional integration of newborn neurons. Exercise may attenuate inflammatory signaling, support angiogenesis and blood–brain barrier repair, and improve mitochondrial biogenesis and quality control; however, much of the mechanistic evidence remains preclinical, and increases in early neurogenic markers do not by themselves establish functional neuronal integration or causality. Current clinical evidence supports cognitive benefits of exercise more strongly than it supports AHN as the indispensable mediator of those benefits. A productive-neurogenesis framework therefore provides a more rigorous basis for interpreting existing studies and designing future experiments that combine lineage tracing, temporally controlled neurogenesis ablation, circuit-level analysis, and domain-specific cognitive outcomes.

Streptococcus pneumoniae is a major cause of pneumonia and other diseases. First-line antibiotics work for infections; however, drug-resistant strains complicate their control. Traditional Chinese medicine (TCM) may help treat drug-resistant infections, although the underlying mechanism is unclear. This study explored the efficacy and mechanism of action of Yinhuapinggan Granules (YHPG) in infected mice, focusing on the PI3K/AKT and RAS/MAPK pathways. A mouse model of Streptococcus pneumoniae infection was established using an intranasal instillation approach. Different doses of Yinhuapinggan granules were then continuously administered for six days after infection. Hematoxylin–eosin (HE) staining, immunohistochemical staining, quantitative polymerase chain reaction (qPCR) analysis and western blotting were used to assess the effects of Yinhuapinggan Granules on the histopathological changes in the lungs, the levels of inflammatory factors and the expression of relevant signaling pathway proteins in mice. YHPG reduces lung damage, inflammation and neutrophil infiltration in mice. These findings suggest that the anti-inflammatory and immunomodulatory effects of YHPG are associated with the observed phenomena. YHPG inhibited the upregulation of streptococcus pneumoniae-induced pathway-associated signaling proteins (IκBα, JNK, P38 and ERK), which may be related to the regulation of the PI3K/AKT and RAS/MAPK signaling pathways. The findings of this study provide experimental evidence for the efficacy and anti-inflammatory mechanism of YHPG against pneumonia caused by Streptococcus pneumoniae infection. The present study confirms that YHPG particles can exhibit therapeutic effects in Streptococcus pneumoniae-infected mice by modulating the PI3K/AKT and Ras/MAPK pathways.

BACKGROUND:Yangyin Tongnao (YYTN) granule is a classic traditional Chinese medicine (TCM) preparation known for its therapeutic efficacy in promoting blood circulation, resolving stasis, nourishing yin, and reinforcing qi. Its neuroprotective effects have been validated through both clinical and preclinical studies. METHODS:A middle cerebral artery occlusion (MCAO) rat model was established, and rats were randomly assigned to five groups: Sham, MCAO, YYTN, Ag490 (JAK2 inhibitor), and Ag490 + YYTN. YYTN was administered intragastrically at 1.73 g/kg/day, while Ag490 was administered via lateral ventricle injection at a volume of 10 μL per rat. Neurological function was assessed using Garcia scores, and infarct volume was determined by TTC staining. Neuronal integrity and damage were evaluated through HE and Nissl staining. Plasma levels of IL-18 and TNF-α were measured via ELISA, and the mRNA expression of IL-6 and IL-1β was quantified by RT-qPCR. Inflammatory markers, including p-JAK2, p-STAT3, NLRP3, ASC, cl-Caspase-1, and cl-IL-1β, were assessed by Western blotting. The distribution of p-JAK2 and p-STAT3 in brain sections was visualized via immunofluorescence staining. RESULTS:Treatment with YYTN significantly improved neurological function, reduced infarct volumes, and mitigated histopathological damage in the brain. Furthermore, YYTN significantly decreased plasma levels of IL-18 and TNF-α. In brain tissue, YYTN downregulated the mRNA expression of IL-6 and IL-1β and reduced key inflammatory proteins, including p-JAK2, p-STAT3, NLRP3, ASC, cl-Caspase-1, and cl-IL-1β. CONCLUSION:These findings suggest that YYTN mitigates cerebral ischemia-reperfusion injury (CIRI)-induced neuroinflammation by inhibiting the JAK2/STAT3 signaling pathway.

Myocardial fibrosis is characterized by excessive cardiac fibroblasts (CF) proliferation and extracellular matrix deposition. The p53 tumor suppressor is known to inhibit CF proliferation, while the polycomb protein Bmi1 negatively regulates p53. Ellagic acid (EA), a natural polyphenol, has demonstrated anti-fibrotic potential, but its mechanism of action in the heart remains unclear. This study investigated whether EA reduces myocardial fibrosis by inhibiting Bmi1 expression. A rat model of myocardial fibrosis was established using isoproterenol (ISO), and rat cardiac fibroblasts were treated with TGF-β to induce a pro-fibrotic phenotype in vitro. Echocardiography and histology confirmed that EA treatment improved cardiac function and reduced fibrosis in ISO-induced rats. In vitro, EA significantly inhibited TGF-β-induced CF proliferation, migration, and differentiation into myofibroblasts. Mechanistically, EA upregulated p53 and its downstream target p21, leading to G1/S cell cycle arrest. Molecular docking and dynamics simulations predicted that EA binds to Bmi1, and subsequent experiments showed EA treatment reduced Bmi1 protein levels. Using a Bmi1 inhibitor (PTC-209) and overexpression plasmids, we demonstrated that the anti-fibrotic effects of EA are mediated in part through inhibition of Bmi1, which in turn activates the p53 pathway. Ellagic acid reduces myocardial fibrosis, at least in part, through suppression of Bmi1 expression. This suppression is associated with relief of the inhibitory effect of Bmi1 on p53, leading to p53/p21 pathway activation, cell cycle arrest, and subsequent inhibition of cardiac fibroblasts proliferation, migration, and differentiation. These findings suggest that Bmi1 may represent a novel therapeutic target for EA treatment of myocardial fibrosis.

Background/Objective: Myocardial fibrosis (MF) is a prevalent pathological endpoint in various heart diseases, characterized by extracellular matrix (ECM) dysregulation and oxidative stress. Hyperoside (Hyp) plays a role in regulating cardiac oxidative stress and fibrosis. This study aimed to elucidate whether Hyp regulates isoproterenol (ISO)-induced MF in mice by modulating the GATA4/HIF-1α signaling pathway and reducing oxidative stress. Methods: The binding affinity of Hyp to GATA4 and HIF-1α was assessed through molecular docking and dynamics simulation. The MF model of mice was established by subcutaneous injection of ISO. Cardiac function was measured by echocardiography. Myocardial injury and collagen deposition were examined using H&E and Sirius red staining. Levels of fibrosis markers, oxidative stress indicators, and GATA4/HIF-1α pathway indicators in serum and heart tissue were quantified by ELISA, Western blot, RT-qPCR and flow cytometry. The distribution of myocardial marker proteins was visualized by immunofluorescence and immunohistochemistry. Results: Molecular docking revealed high binding affinity of Hyp to GATA4 and HIF-1α (binding energies < -5.0 kcal·mol-1), and dynamics simulation showed that the complex's structure remained stable over 100 nanoseconds (RMSD < 0.1 nm). High-dose Hyp (36 mg/kg) significantly improved cardiac function, myocardial injury, collagen deposition, and inflammatory infiltration in MF mice. Molecularly, Hyp effectively reduces oxidative stress and fibrosis through upregulating GATA4 and downregulating HIF-1α. Conclusions: Hyp suppresses oxidative stress by activating the GATA4/HIF-1α pathway, presenting a promising therapeutic target for the treatment of MF.

BACKGROUND:Coronary heart disease (CHD) and ischemic stroke (IS) frequently co-occur; however, their shared molecular drivers remain poorly understood, limiting the development of dual-indication therapies. This study aims to characterize the bidirectional genetic relationship between CHD and IS and to prioritize shared candidate proteins as potential therapeutic targets or biomarkers. METHODS:Bidirectional two-sample Mendelian randomization (MR) was performed using GWAS summary statistics, with inverse-variance weighting as the primary analysis method. To resolve conceptual ambiguity regarding protein roles, we integrated MR and colocalization analyses across CHD, IS, and plasma proteomic QTLs to identify proteins driven by shared genetic signals. Candidate proteins were further contextualized using single-cell transcriptomics, followed by drug-repurposing predictions and exploratory molecular dynamics (MD) simulations. In vivo experiments were conducted to provide expression-level validation of prioritized candidates in disease models. RESULTS:IVW MR supported a bidirectional association between IS and CHD (OR = 1.307, 95% CI 1.096-1.560, P = 0.003). Integrative analyses prioritized CYR61, IL2RB, and ST3GAL2 as shared candidate proteins. While MD simulations generated structural hypotheses regarding drug-protein interactions, in vivo experiments confirmed significant differential expression of these candidates, supporting their clinical relevance at the expression level. CONCLUSION:This integrative genomics framework clarifies the shared genetic architecture of IS and CHD and identifies high-priority protein candidates for future functional and pharmacological validation.

Endothelial-mesenchymal transition (EndMT) is increasingly recognized as a critical factor contributing to myocardial fibrosis, which ultimately leads to cardiac dysfunction. Naringenin (NAR), a major flavonoid derived from citrus fruits, exhibits various pharmacological properties, including anti-inflammatory, antioxidant, and cardioprotective effects. However, the pharmacological mechanism by which NAR counteracts EndMT remains unclear. In this study, a heart failure model was established in rats by subcutaneous injection of isoproterenol hydrochloride (ISO, 5 mg/kg for 7 days), and human umbilical vein endothelial cells (HUVECs) were stimulated with angiotensin II (Ang II, 1 µM) in vitro. NAR was administered orally (50 and 100 mg/kg/day) for 28 days after ISO induction. Echocardiography revealed that NAR significantly improved ejection fraction (EF) and fractional shortening (FS), and reduced systolic/diastolic diameters and volumes. Histological analyses (H E and Masson’s trichrome staining) showed that NAR attenuated ISO‑induced myocardial disarray, hypertrophy, and collagen deposition. Immunohistochemistry and Western blot demonstrated that NAR downregulated fibrosis markers (collagen I, collagen III) and the mesenchymal marker α‑SMA, while upregulating the endothelial marker CD31. Mechanistically, NAR increased the phosphorylation of AKT and GSK3β, as well as the expression of β‑catenin, indicating activation of the AKT/GSK3β/β‑catenin pathway. Consistently, the GSK3β inhibitor AR‑A014418 reversed the protective effects of NAR in HUVECs. In summary, our findings demonstrate that naringenin attenuates myocardial fibrosis by inhibiting EndMT via modulation of the AKT/GSK3β/β‑catenin signaling pathway. These results highlight the promising therapeutic potential of naringenin in managing myocardial fibrosis.

Weber syndrome, characterized by ipsilateral oculomotor palsy and contralateral hemiplegia, frequently occurs in ischemic stroke cases and is difficult to recover from. Hydroxysafflor yellow A (HSYA), a bioactive component present in Carthamus tinctorius L. and the standardized preparation Danhong injection, has shown protective effects in animal models of various neurological diseases. This research was conducted to evaluate the therapeutic potential of HSYA in ischemic stroke-associated Weber syndrome, while elucidating its mechanistic basis. A rat MCAO model was induced to detect the effects of HSYA on motor dysfunction, ipsilateral ptosis and neuronal death in the right cerebral peduncle. By western blot, immunohistochemistry staining and immunofluorescence staining, we explored the involvement of oxidative stress and DNA damage in HSYA's neuroprotective action. HSYA treatment for consecutive 7 days significantly improved neurological function, grip strength, and asymmetry of bilateral eye clefts in CI/R-injured rats. HSYA also reduced cerebral infarction, preserved neuronal survival, and mitigated histopathological damage in the cerebral peduncle. Mechanistically, HSYA alleviated oxidative stress by preserving CAT, GSH, and SODM levels while inhibiting iNOS overexpression. Furthermore, CI/R injury triggered substantial DNA damage in the cerebral peduncle, as indicated by upregulated levels of 53BP1 and γ-H2A.X. Contents of PARP1, AIF, and MIF were also significantly elevated, accompanied by obvious upregulation of apoptotic cell death, while HSYA treatment effectively attenuated these deleterious effects. HSYA protects against CI/R injury and associated Weber syndrome, and the mechanism involves suppressing oxidative stress and limiting DNA injury.

OBJECTIVE:This study aimed to investigate the antimicrobial and anti-inflammatory effects of Yinhuapinggan Granules (YHPG) in combination with cefotaxime (CTX) against extended-spectrum beta-lactamase-producing Escherichia fergusonii (ESBL-EF), focusing on how YHPG enhances antibiotic efficacy and modulates inflammatory pathways associated with acute lung injury (ALI). METHODS:The antimicrobial activity of YHPG and CTX was assessed by measuring the minimum inhibitory concentration (MIC) of CTX in the presence of YHPG. qRT-PCR was used to detect the expression of bacterial resistance genes, including acrA, acrB, tolC, OXA-2, TEM-1, and outer membrane porins ompC and ompF. Outer membrane permeability was evaluated using AKP and Bradford assays, while biofilm disruption was analyzed by laser confocal microscopy. In vivo, the effects of YHPG were evaluated in a mouse pneumonia model induced by ESBL-EF infection, including lung index, histopathological changes, and inflammatory cytokine levels. Inflammatory cytokine levels were measured in serum, and the expression of key genes in the NLRP3 inflammasome pathway was analyzed by qRT-PCR. RESULTS:The combination of YHPG and CTX significantly inhibited the growth of ESBL-EF and enhanced its outer membrane permeability. Additionally, YHPG modulated the expression of key bacterial resistance genes, including acrA, acrB, tolC, OXA-2, and TEM-1, as well as the outer membrane porins ompC and ompF. YHPG also effectively disrupted the biofilm structure of ESBL-EF, reducing its protective barrier. In vivo, YHPG treatment alleviated pulmonary inflammation and tissue damage in mice. Serum levels of pro-inflammatory cytokines, including TNF-α, IL-6, and IL-1β, were significantly reduced, indicating a potent anti-inflammatory effect. Furthermore, YHPG inhibited the activation of the NLRP3-ASC-CASP1-GSDMD signaling pathway, further mitigating inflammation and tissue damage associated with ESBL-EF infection. CONCLUSION:YHPG enhances the antibacterial activity of CTX against ESBL-EF by increasing membrane permeability, disrupting biofilms, and modulating the inflammatory response. This study suggests that YHPG could serve as an adjunctive treatment for antibiotic-resistant infections, offering a promising approach to combat bacterial resistance and inflammation associated with ESBL-EF infections.

Astragalus mongholicus Bunge [Fabaceae; Astragali radix] is an herb widely used in traditional Chinese medicine. It has diuretic, anti-aging, antihypertensive, immune-boosting, liver-protective, anti-stress and other extensive pharmacological effects. In recent years, Astragalus and its extract have been used to treat lung and stomach qi deficiency as well as general qi deficiency. This paper summarizes the mode of action and mechanisms of Astragalus in treating various diseases, and provides valuable insights for the future application, development, and improvement of Astragalus. In this paper, literature on the use of Astragalus in treating related diseases over the past five years was collected from PubMed and CNKI databases, and the pathogenic mechanisms of Astragalus and its extracts were reviewed. Its mechanism of action is primarily involved in antioxidant protection, anti-inflammatory effects, and anti-apoptotic properties. This provides a new research direction for future studies and clinical treatments using Astragalus.

BACKGROUND:The relationship between hematocrit (HCT) levels and the occurrence of major adverse cardiovascular events (MACEs) in patients with acute myocardial infarction (AMI) remains unexplored. A better understanding of this interplay may enhance the prognosis and management of AMI patients. METHODS:Between January 2021 and August 2022, clinical data were collected from patients diagnosed with AMI at 10 tertiary healthcare institutions in China. A total of 1946 eligible participants were included and divided into three groups based on sex-specific tertiles of HCT levels upon admission: 648 patients with low HCT levels, 649 patients with intermediate HCT levels, and 649 patients with high HCT levels. Follow-up approaches included hospital outpatient visits, inpatient stays, and telephone calls for 180 days. The primary endpoint was the occurrence of MACEs. Influential factors, including general information, admission status, and supplementary examination results that differed across the cohorts, were analyzed. Cox regression analysis was employed to evaluate the 180-day MACE rates and HCT levels in patients with AMI. To assess the reliability of the findings, three sensitivity analyses and subgroup analyses were performed. RESULTS:During this time, 136 individuals in the low HCT group, 77 in the intermediate HCT group, and 73 in the high HCT group experienced endpoint events. With all covariates controlled, the Cox regression analysis indicated that the low HCT group had a higher risk of MACEs compared to the intermediate HCT group [hazard ratio (HR) = 1.44, 95% confidence interval (CI) = 1.07-1.95, p = 0.017]. The low HCT group also presented a higher risk of acute coronary syndrome (HR = 1.57, 95% CI = 1.06-2.32, p = 0.024). However, the high and intermediate HCT groups exhibited comparable prognoses for AMI. The limited cubic spline plot revealed that HCT values between 41.58% and 45.36% implied a protective effect against MACEs. These results were further verified by sensitivity analysis, and the subgroup analysis showed no variable interaction. CONCLUSIONS:Our findings indicate that low HCT levels in patients with AMI increase the incidence of MACEs within 180 days, offering new insights into the prognosis and management of AMI patients. CLINICAL TRIAL REGISTRATION:ChiCTR2200066456.

Ischemic stroke (IS) initiates complex systemic responses that extend beyond focal brain injury. To capture these multifaceted changes, proteomics, untargeted metabolomics, and lipidomics are integrated from IS patients spanning a range of clinical severities. This multi-omics approach reveals distinct molecular subtypes characterized by immune activation, oxidative stress, and metabolic dysregulation. Notably, elevated levels of migrasomes are identified in patient plasma and mouse brain tissue. Proteomic profiling of these migrasomes shows enrichment in complement, coagulation, and cholesterol-associated pathways. Functional assays further demonstrate that migrasomes derived from peripheral immune cells exacerbate ischemic injury and intensify post-stroke inflammation. Together, these findings position migrasomes as critical drivers of IS pathophysiology and highlight them as promising targets for therapeutic intervention.

Chronic inflammation linked to atherosclerosis is closely related to a trained immunoregulatory network. Traditional studies primarily focus on the pro-inflammatory memory of monocytes, they frequently neglect important aspects such as the cell’s plasticity, interactions between different organs, and the dynamic regulation of the metabolism-vascular axis. This review presents four novel frameworks, including the trained immunity plasticity spectrum model. It demonstrates how monocytes maintain a dynamic balance between pro-inflammatory, tolerogenic, and anti-inflammatory phenotypes, regulated by mTOR/AMPK signaling and competitive histone modifications. The trained immunity–metabolism–vascular axis shows that metabolic disorders can change the way immune memory is formed. They achieve this by modifying the vascular microenvironment through epigenetic changes, exosomes, and products of mitochondrial stress. The cross-organ trained immunity framework reveals how remote epigenetic communication between the bone marrow, gut, and liver influences the development of monocytes. Finally, dynamic immune reprogramming integrates CRISPR-based epigenetic editing, metabolism-focused interventions, and AI-driven multi-omics predictions. This approach signifies a major transition from simply alleviating symptoms to accurately reshaping immune memory. This review reinterprets the immunometabolic mechanisms of atherosclerosis. It also lays the foundation for personalized therapies enhanced by AI and explores new interdisciplinary research avenues.

Helper T cells 17 (Th17) and their effector cytokine, interleukin-17A (IL-17A), play a dual role in immune homeostasis. On one hand, they are essential in defense against extracellular pathogens, such as bacteria and fungi, by inducing chemokine production and recruiting neutrophils. On the other hand, their dysregulated activity is strongly linked to autoimmune and inflammatory disorders, including multiple sclerosis, Alzheimer’s disease, Parkinson’s disease, and others. This article reviews the molecular mechanisms regulating Th17 differentiation and function, emphasizing the role of transcription factors like RORγt and RORα, as well as the influence of cytokines such as IL-6, IL-23, and TGF-β. Additionally, it explores the imbalance between pro-inflammatory Th17 cells and regulatory T cells (Tregs), a critical axis in the pathogenesis of autoimmune and neuroinflammatory diseases. In the context of neurological disorders, Th17 cells can infiltrate the central nervous system (CNS), where they contribute to neuroinflammation by activating microglia and astrocytes, exacerbating damage in conditions such as multiple sclerosis, traumatic brain injury, and neurodegenerative diseases. Emerging therapies, including anti-IL-17 monoclonal antibodies and natural modulators, are discussed as potential strategies to restore the Th17/Treg balance without compromising protective immunity. Finally, the need for further research is highlighted to elucidate the specific mechanisms of Th17 infiltration into the CNS, their interaction with the gut microbiota, and the development of personalized therapies. The integration of immunological, metabolic, and environmental approaches offers promising perspectives for the treatment of Th17/IL-17-mediated diseases.

ICH, a severe stroke, causes neuronal death, neuroinflammation, and cerebral edema due to mitochondrial and immune dysfunction. The molecular mechanisms of secondary brain injuries are unclear, limiting effective therapies. This study used bioinformatics and experiments to explore miR-940’s role in ICH. MitoDEGs were identified via MitoCarta 3.0, and key miRNAs predicted using TargetScan and miRDB. A collagenase-induced ICH rat model with antagomir knockdown was used for validation. Neuronal damage, mitochondrial proteins, and immune cell dynamics were assessed using histopathology, qRT-PCR, Western blotting, and flow cytometry. Bioinformatics identified seven MitoDEGs and miR-940. Functional analysis linked them to mitochondrial metabolism and neuroinflammation. Inhibiting miR-940 in vivo reduced neuronal apoptosis and cerebral edema, and reversed RHOBTB1 and BCL2A1 dysregulation. Immune profiling showed increased monocyte/NK cell infiltration and decreased T/B lymphocytes in ICH, correlating with MitoDEGs. Flow cytometry confirmed miR-940’s role in restoring T/B cell homeostasis, and Western blotting validated key MitoDEG expression changes. This study establishes miR-940 as a key regulator of mitochondrial-immune crosstalk in ICH, modulating neuronal survival and immune microenvironment remodeling. The identified MitoDEGs and miR-940 axis may serve as potential diagnostic and therapeutic targets for ICH.

Penicillin-resistant Streptococcus pneumoniae (PRSP), for which novel treatment medicines are required, has expanded extensively due to the overuse of antibiotics. This study aimed to detect the optimal ratio of the combination of the main components based on Yinhuapinggan granules (YHPG) to generate novel treatment concepts for PRSP-induced lung injury. Three representative main components: chlorogenic acid (C), amygdalin (A), and puerarin (P) were selected, and the optimal combination of these three components was determined by an orthogonal experiment. Investigations were conducted on the potential mechanisms underlying the protective effect of this optimized combination against PRSP-induced lung epithelial cell damage. Meanwhile, the bacteriostatic effect was further explored through the optimized combination of these natural products combined with penicillin G (PG). The optimized combination CAP (C: 16 µg/mL, A: 24 µg/mL, P: 24 µg/mL) screened by the orthogonal experimental design reduced cell damage in a model of human lung epithelial cells infected by PRSP, and the combination of CAP and PG had a synergistic effect. At the cellular level, CAP attenuated lung epithelial cell injury by modulating the TLRs/MyD88 inflammatory pathway. At the bacterial level, CAP modulated the virulence and drug resistance of PRSP, resulting in enhanced bacterial inhibition by the combination of CAP and PG. Taken together, our results suggest that CAP can modulate or synergize with PG to modulate the TLRs/MyD88 pathway and attenuate PRSP-induced lung injury, and can be used as a potential drug for treating PRSP infection.

Ischemic stroke (IS) is a highly complex and heterogeneous disease involving multiple pathophysiological events. A better understanding of the pathophysiology of IS will enhance preventive, diagnostic and therapeutic strategies. Despite significant advances in modern medicine, the molecular mechanisms of IS are still largely unknown. The high-throughput omics approach opens new avenues for identifying IS biomarkers and elucidating disease pathogenesis mechanisms. Single-cell omics enables a more thorough and in-depth analysis of the cellular interactions and properties in IS. This will lead to a better understanding of the onset, treatment and prognosis of IS. In this paper, we first reviewed the disease signatures and mechanisms research of IS. Subsequently, the use of single-cell omics to comprehend the mechanisms of IS was discussed, along with some recent developments in the field. To further delineate the upstream pathogenic alterations and downstream molecular impacts of IS, we also discussed the current use of machine learning approaches to single-cell omics data analysis. Particularly, single-cell omics is being used to inform risk assessment, early patient diagnosis and treatment strategies, and their potential impact on precision medicine. Thus, we summarized the role of single-cell omics in precision medicine. Despite the relative youth of the field, the development of single-cell omics promises to provide a powerful tool for elucidating the pathogenesis of IS.

BACKGROUND:Intracerebral hemorrhage (ICH) is a devastating neurological disorder characterized by oxidative stress, inflammatory cascades, and metabolic dysregulation. Ginsenoside Rd (G-Rd) and chrysophanol (Chr), two natural compounds with antioxidative and anti-inflammatory properties, have demonstrated neuroprotective potential, however, their mechanisms in ICH remain unclear. OBJECTIVE:This study aimed to investigate the protective effects of G-Rd and Chr against heme-induced injury in HT22 cells and in a rat model of ICH, with a focus on oxidative stress, inflammation, apoptosis, and metabolic regulation. METHODS:In vitro, HT22 cells were exposed with heme (10 μmol/L, 12 h) to simulate ICH injury, followed by treatment with G-Rd and Chr (80 μmol/L). Reactive oxygen species (ROS), apoptosis, mitochondrial membrane potential, and inflammatory cytokines (TNF-α, IL-1β, IL-6, IL-10) were assessed using flow cytometry, fluorescence microscopy, and ELISA. In vivo, ICH was induced in rats via collagenase injection. Neurological function, hematoma volume, histopathology, and metabolic enzymes (SOD, MDA, NAD+, Ca2+-ATPase) were evaluated. Western blotting was used to analyze key enzymes in the serine-glycine-one‑carbon (SGOC) pathway. RESULTS:G-Rd and Chr significantly suppressed ROS production (P < 0.05), downregulated pro-inflammatory cytokines (TNF-α, IL-1β, IL-6; P < 0.01), and inhibited apoptosis (P < 0.01) in HT22 cells. Both compounds restored mitochondrial membrane potential and alleviated cellular damage. In the ICH rat model, combined treatment improved neurological scores by 45 % (P < 0.01), decreased hematoma volume by 38 % (P < 0.01), and restored metabolic homeostasis through modulation of SGOC pathway enzymes (PHGDH, PSAT1, PSPH, SHMT2; P < 0.05). Synergistic effects were observed in both hematoma resolution and neuroprotection. CONCLUSION:G-Rd and Chr confer neuroprotection in ICH through antioxidative, anti-inflammatory, anti-apoptotic, and metabolic regulatory mechanisms. Their synergistic efficacy underscores their promise as therapeutic candidates, meriting further investigation of their molecular targets and translational potential.

Background and aimThe NLRP3 inflammasome can be activated after intracerebral hemorrhage (ICH), triggering an inflammatory response in the brain. Chinese herbal medicine Zhongfeng Xingnao Prescription (ZFXN) is commonly used in China for intracerebral hemorrhage treatment. However, the underlying treatment mechanism of it is unclear. The purpose of our study is to investigate whether ZFXN alleviates injury after intracerebral hemorrhage by blocking the activation of CaMKII/NF-κB p65/NLRP3/GSDMD signaling axis.Experimental procedureWe evaluated the protective effect of ZFXN in a rat model of collagenase Ⅶ‐induced ICH. The neurological deficit score, cerebral hematoma-injury ratio, pathology, and ultrastructure of tissue surrounding the hematoma were evaluated after 5 days of ZFXN treatment, CaMKII/NF-κB p65/NLRP3/GSDMD signaling axis-related protein expression around the hematoma was assessed by Western blot and immunohistochemistry. Meanwhile, ELISA measured the levels of IL-1β, IL-18, IL-6 and TNF-α in serum.Results and conclusionAfter 5 days of ZFXN treatment, the score of neurological deficit and hematoma damage ratio decreased, and the cell destruction such as edema and vacuole conditions around the hematoma improved. The mechanism investigation results showed that ZFXN down-regulated expressions of CaMKII/NF-κB p65/NLRP3/GSDMD signaling axis-related protein around the hematoma area. In addition, ZFXN could attenuate the inflammatory response by regulating the activation of NLRP3 inflammasome after ICH. For the first time, we found that the efficacy of ZFXN on ICH might be related to the regulation of NLRP3 inflammasome.

PANoptosis is a newly identified form of cell death that encompasses pyroptosis, apoptosis, and necroptosis. Numerous studies have highlighted the significance of PANoptosis in brain ischemia-reperfusion (I/R) injury. Calycosin, a natural product with diverse biological activities, has demonstrated a significant reduction in neuronal death caused by ischemic brain injury by modulating multiple cell death pathways. In order to investigate the potential mechanisms underlying the neuroprotective role of calycosin in alleviating PANoptosis-induced damage in ischemic stroke therapy, we used mouse hippocampal neuronal cell line HT22 to stimulate ischemia in vitro through Oxygen and Glucose Deprivation/Reperfusion (OGD/R) and established molecular docking to assess the binding affinity of Calycosin with key targets and molecular dynamics simulations (MDS) to study the stability of the ligand-protein complex. The results demonstrate that Calycosin could improve the cell growth of HT22, leading to enhanced cell viability, reduced lactate dehydrogenase leakage, and decreased cell apoptosis after OGD/R. It also regulated the expression of PANoptosis-related genes such as NLRP3, GSDMD, MLKL, and RIPK1 and increased the Bcl-2/Bax ratio, effectively reducing cellular damage and providing protection. Molecular docking and MDS simulations demonstrated strong binding activity and stability between Calycosin and PANoptosis-related targets. Furthermore, Calycosin successfully passed the drug similarity (DS) evaluation and exhibited favorable absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties and biological activity. In conclusion, Calycosin could alleviate ischemic stroke by inhibiting PANoptosis, reducing neuronal inflammation and apoptosis, and improving damage caused by the OGD/R. Thus, it could serve as a potential therapy for ischemic stroke.