BACKGROUND:Cerebral ischemia-reperfusion injury (CIRI) is a critical pathological process following ischemic stroke, with ferroptosis being increasingly recognized as a key contributor to neuronal damage. However, the regulatory mechanisms, particularly the role of specific transcription factors like E26 transformation-specific variant 4 (ETV4), remain poorly understood. This study aimed to investigate the function and underlying mechanism of ETV4 in neuronal ferroptosis during CIRI. METHODS:In vitro, SH-SY5Y cells subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) were used to model CIRI. Cell viability and ferroptosis markers (Fe2 +, malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), reactive oxygen species (ROS)) were assessed. Molecular expressions were measured by Real-time Quantitative PCR (RT-qPCR) and/or western blot. In vivo, a rat model of middle cerebral artery occlusion/reperfusion (MCAO/R) was established, and brain injury was evaluated via 2% solution of 2,3,5-triphenyl tetrazolium chloride (TTC), hematoxylin-eosin (HE), and TdT-mediated dUTP nick-end labeling (TUNEL) staining. Chromatin immunoprecipitation (ChIP), dual-luciferase reporter, RNA pull-down, RNA immunoprecipitation (RIP), and actinomycin D assays were employed to validate the molecular interactions within the ETV4/Y-box binding protein 1 (YBX1)/glutathione peroxidase-4 (GPX4) axis. RESULTS:ETV4 was significantly down regulated in both the MCAO/R model and OGD/R-treated cells. Overexpression of ETV4 markedly attenuated OGD/R-induced oxidative stress, ferroptosis in vitro, and ameliorated brain injury in vivo. Mechanistically, ETV4 transcriptionally activated YBX1 by directly binding to its promoter. YBX1, in turn, stabilized GPX4 mRNA, which was modified by NSUN2-mediated methylation. Crucially, the protective effects of ETV4 in vitro were abolished upon YBX1 or GPX4 knockdown. CONCLUSION:Our findings demonstrate that ETV4 transcriptionally up regulates YBX1 to stabilize GPX4 mRNA in an NSUN2-m5Cmethylation dependent manner, thus suppressing neuronal ferroptosis. This reveals a novel ETV4/YBX1/GPX4 axis as a potential therapeutic target for CIRI.

BACKGROUND:Ischemic stroke (IS) is an acute cerebrovascular disease characterized by high morbidity and mortality, with limited current treatment options. Tumor protein p53-inducible nuclear protein 2 (Tp53inp2) is known to be a positive regulator of autophagy under physiological conditions, but the mechanism of Tp53inp2 in IS remains unclear. In this study, we aimed to explore the mechanism of Tp53inp2 in IS. METHODS:Primary neural stem cells (NSCs) were extracted and identified. An OGD/R cell model was constructed. Tp53inp2 was knocked down and rapamycin was added. A middle cerebral artery occlusion (MCAO) animal model was constructed, and then 5 μL of 5 × 105 NSCs, either untreated or transfected with sh-NC or sh-Tp53inp2, were injected. Additionally, at the cellular level, Ptgs2 or Tp53inp2 was overexpressed, and METTL14 was knocked down. RESULTS:Inhibition of Tp53inp2 mitigated OGD/R-induced mitophagy and ROS levels in vitro. Moreover, inhibition of Tp53inp2 mediated neuronal differentiation of OGD/R treated NSCs by suppressing mitophagy. At the animal level, the transplantation of NSCs with a knockdown of Tp53inp2 increased neuronal differentiation, thereby alleviating the effects of MCAO and mitigating cognitive impairments in MCAO model mice. Ptgs2 was further screened and validated as a downstream target mediating the effect of Tp53inp2 on NSCs. At the cellular level, Tp53inp2 alleviated OGD/R-induced mitophagy and ROS levels by regulating Ptgs2 expression. METTL14 could regulate Tp53inp2 expression by modulating the functional m6A modification sites on Tp53inp2 mRNA. Inhibition of METTL14 alleviated OGD/R-induced mitophagy and the rise of ROS levels in NSCs by inhibiting the Tp53inp2/Ptgs2 axis. CONCLUSIONS:Inhibition of METTL14 alleviated OGD/R-induced neuronal differentiation injury in NSCs by inhibiting the Tp53inp2/Ptgs2 axis. By elucidating the mechanisms involving Tp53inp2, Ptgs2, and METTL14, this research offers a foundation for developing new strategies to enhance neuronal differentiation and mitigate cognitive impairments in stroke patients.

BACKGROUND:Cerebral ischemia-reperfusion (CI/R) injury, a major complication of ischemic stroke, is characterized by mitochondrial dysfunction and neuronal apoptosis, and understanding its underlying molecular mechanisms is essential for the development of effective therapeutic strategies. This study aimed to investigate the role of ubiquitin-specific protease 7 (USP7) in CI/R injury and elucidate its regulatory mechanisms. METHODS:A rat model of middle cerebral artery occlusion/reperfusion (MCAO/R) and an in vitro neuronal model subjected to oxygen-glucose deprivation/reperfusion (OGD/R) were used to mimic CI/R injury. USP7 was overexpressed or knocked down, with or without co-treatment, using the autophagy inhibitor 3-methyladenine (3-MA). Neurological function was evaluated using standardized scoring systems, and cerebral infarct volume was quantified by TTC staining. Histopathological changes in the cortex and hippocampus were assessed using hematoxylin-eosin (HE) and Nissl staining. Neuronal viability and apoptosis were measured by CCK-8 assay, TUNEL staining, and flow cytometry. To assess cellular metabolism and oxidative stress, ATP and LDH levels, along with antioxidant markers (including SOD, GSH, and GSH-Px), were analyzed using commercial biochemical kits. Mitochondrial morphology and autophagosome formation were visualized using transmission electron microscopy. Gene and protein expression levels were quantified by qRT-PCR and Western blotting, respectively. Immunofluorescence microscopy was performed to evaluate the subcellular localization of target proteins and co-localization with mitochondrial membrane markers. Lastly, protein-protein interactions and ubiquitination modification were analyzed by co-immunoprecipitation assays. RESULTS:USP7 overexpression significantly alleviated neurological deficits, reduced infarct volume, attenuated histological damage, and decreased neuronal apoptosis in the MCAO/R model. Similarly, in the OGD/R model, USP7 overexpression markedly enhanced neuronal viability, suppressed apoptosis, restored ATP production, improved antioxidant capacity (as evidenced by increased levels of SOD, GSH, and GSH-Px), and reduced LDH release. Mechanistically, USP7 stabilized SIRT1 protein expression through deubiquitination, which in turn activated the PINK1/Parkin pathway and enhanced mitophagy. This activation was demonstrated by an increased LC3II/LC3I ratio, elevated ATG5 expression, enhanced co-localization of Tomm20 and Parkin, and increased autophagosome formation. Moreover, these protective effects were abolished when either 3-MA treatment was applied or SIRT1/PINK1 expression was knocked down. CONCLUSION:USP7 mitigates CI/R injury by promoting PINK1/Parkin-dependent mitophagy through SIRT1 deubiquitination and stabilization, suggesting USP7 as a potential therapeutic target for ischemic stroke.

Ischemic stroke (IS) is known for its high morbidity, disability and mortality rates, and studies designed to explore its pathophysiological mechanisms and identify novel therapeutic strategies are urgently needed. We aimed to probe the effects of the deubiquitinase OTUD3-IRP2-p53/PTGS2 pathway on cerebral ischemia‒reperfusion (I/R) injury and hippocampal neuron ferroptosis. A cerebral I/R mouse model was established. Furthermore, lentiviral vectors that overexpressed OTUD3 and knocked down IRP2 were constructed, and a series of assays were performed to probe the OTUD3/IRP2/p53/PTGS2 mechanism. An oxygen‒glucose deprivation and reoxygenation (OGD/R) model of mouse hippocampal neurons was constructed. Then, OTUD3 and IRP2 were knocked down and overexpressed, and p53 was overexpressed to explore the mechanism of the OTUD3/IRP2/p53/PTGS2 pathway. OTUD3 and IRP2 were expressed at low levels in cerebral I/R models. OTUD3 promoted IRP2 expression to protect damaged hippocampal neurons. Moreover, IRP2 affected ferroptosis in hippocampal neurons. In addition, IRP2 inhibited p53. After IRP2 and p53 were overexpressed, IRP2 regulated the p53/PTGS2 pathway and affected ferroptosis in hippocampal neurons. In vivo, after overexpressing OTUD3 and knocking down IRP2, we found that overexpression of OTUD3 promoted IRP2 expression to reduce ferroptosis in hippocampal neurons and improve cerebral I/R injury via the inhibition of the p53/PTGS2 pathway. The deubiquitinase OTUD3 stabilized IRP2 expression to reduce hippocampal neuron ferroptosis via the p53/PTGS2 pathway to subsequently ameliorate cerebral I/R injury.

OTU domain-containing protein 3 (OTUD3) is a crucial deubiquitinase that exhibits significant expression differences across various disease models. OTUD3 plays a role in regulating biological functions such as apoptosis, inflammatory responses, cell cycle, proliferation, and invasion in different cell types. By deubiquitinating key substrate proteins, OTUD3 is involved in essential physiological and pathological processes, including innate antiviral immunity, neural development, neurodegenerative diseases, and cancer. OTUD3 exhibits tumor-suppressive effects in breast cancer, esophageal cancer, colon cancer, and papillary thyroid cancer, but acts as an oncogenic in liver and lung cancers. OTUD3 serves as a biomarker in predicting diagnosing, and assessing prognosis for certain malignancies. Despite its potential, the molecular mechanisms of OTUD3 in many diseases are still not well-understood, and exploring OTUD3's regulatory mechanisms is essential for comprehending its roles in immunity and disease. Future research will focus on developing OTUD3-targeted therapies.

Introduction: The purpose of this study was to investigate the effects of bone marrow mesenchymal stem cells (BMSCs) exosomal miR-345-3p and tumor necrosis factor receptorassociated factor 6 (TRAF6) on cerebral ischemia reperfusion (CIR) injury. Exosomes (Exos) derived from BMSCs were isolated and identified. PC12 (rat pheochromocytoma) cells were used to establish an oxygen and glucose deprivation/reoxygenation (OGD/R) model. Methods: Cell counting kit-8, TUNEL staining, lactate dehydrogenase staining, RT-qPCR, and western blotting were utilized for analyzing the functions of miR-345-3p about PC12 cells. Dualluciferase reporter experiment was then to confirm the link between miR-345-3p and TRAF6. Finally, using male SD rats, the middle cerebral artery occlusion (MCAO) model was constructed. Regulation of I/R damage in MCAO rats of miR-345-3p and TRAF6 were further explored in the changes of modified neurological severity score, cerebral infarction pictures, relative infarct volume, and histopathological changes. After OGD/R treatment, neuronal apoptosis was dramatically increased. After treatment with exosomal miR-345-3p, OGD/R-induced neuroapoptosis was dramatically inhibited. Exosomal miR-345-3p inhibited OGD/R-induced neuroapoptosis by downregulating the expression of TRAF6. However, the miR-345-3p inhibitor aggravated the changes caused by OGD/R. Results: The corresponding regulations of miR-345-3p were reversed with TRAF6 overexpression. The animal experiments in vivo further verified that miR-345-3p ameliorated brain I/R injury in MCAO rats by targeting TRAF6. Conclusion: This study found that BMSCs-exosomal miR-345-3p protected against CIR injury by decreasing TRAF6.

Mesenchymal stem cells (MSCs)-derived exosomes are demonstrated to exert neuroprotective effects in stroke. We aimed to explore the role and mechanism of long non-coding RNA (lncRNA) KLF3 antisense RNA 1 (KLF3-AS1) in bone marrow mesenchymal stem cells-derived exosomes (BMSCs-Exos) in cerebral ischemia/reperfusion (I/R) injury. Exosomes were isolated from the culture medium of BMSCs. A mouse model of middle cerebral artery occlusion (MCAO) in vivo and a BV-2 cell model of oxygen and glucose deprivation/reoxygenation (OGD/RX) in vitro were established. Cell viability and apoptosis were detected using MTT assay, TUNEL staining and flow cytometry, respectively. Related proteins were determined with western blot and immunohistochemistry, while related RNAs were analyzed by RT-qPCR. Neurological deficit and cerebral infarct volume were evaluated by the modified neurological severity score (mNSS) and TTC staining, respectively. Our observations indicate that exosomes derived from BMSCs-preconditioned medium exerted neuroprotective effects, as indicated by the increased cell viability and the suppressed apoptosis in OGD/RX-suffered BV-2 cells. KLF3-AS1 expression was upregulated in BMSCs-Exos. Furthermore, KLF3-AS1 knockdown antagonized the protective effects of BMSCs-Exos. Mechanistically, BMSCs-Exos carrying KLF3-AS1 inhibited apoptosis via enhancing autophagy. KLF3-AS1 was found to recruit ETS variant transcription factor 4 (ETV4), which upregulated Sirt1 expression. Knockdown of KLF3-AS1 neutralized the protective effects of BMSCs-Exos on MCAO-induced brain injury, which was then reversed by the treatment with Sirt1 inhibitor EX527. We concluded that KLF3-AS1 derived from BMSCs-Exos promoted autophagy to alleviate I/R injury via ETV4/Sirt1 axis.

This report aimed to explore whether miR-188-5p regulated the pathological regulatory network of cerebral ischemia/reperfusion (I/R) injury. We simulated the cerebral I/R injury model with MACO/R and OGD/R treatments. Neuronal viability and apoptosis were assessed. The contents of miR-188-5p and Lin 28a were evaluated. The abundances of apoptosis-related proteins (Bax, Bcl-2, and cleaved caspase-3) and pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) were measured. The interaction of miR-188-5p and Lin28a was confirmed. Lin28a silencing was supplemented to determine the delicate regulation of miR-188-5p. We revealed that miR-188-5p was upregulated and Lin28a was downregulated in I/R rats and OGD/R-induced cells. miR-188-5p silencing remarkably reduced the cerebral infarction volume, neurobehavioral score, brain edema, and Evans blue leakage. miR-188-5p silencing enhanced neuronal viability and alleviated apoptosis. The abundance of Bax and cleaved caspase-3 was reduced by miR-188-5p silencing, while Bcl-2 was augmented. miR-188-5p silencing impeded the contents of TNF-α, IL-1β, and IL-6. miR-188-5p interacted with Lin28a and negatively regulated its expression. Interestingly, extra Lin28a silencing reversed apoptosis and the content of inflammatory cytokines. Our studies confirmed that miR-188-5p silencing alleviated neuronal apoptosis and inflammation by mediating the expression of Lin28a. The crosstalk of miR-188-5p and Lin28a offered a different direction for ischemic stroke therapy.

The objective of this research was to investigate the role of lncRNA MALAT1 and HSP90 in the regulation of neuronal necroptosis in mice with cerebral ischemia-reperfusion (CIR). We used male C57BL/6J mice to establish a middle cerebral artery occlusion (MCAO) model and conducted in vitro experiments using the HT-22 mouse hippocampal neuron cell line. The cellular localization of NeuN and MLKL, as well as the expression levels of neuronal necroptosis factors, MALAT1, and HSP90 were analyzed. Cell viability and necroptosis were assessed, and we also investigated the relationship between MALAT1 and HSP90. The results showed that MALAT1 expression increased after MCAO and oxygen-glucose deprivation/re-oxygenation (OGD/R) treatment in both cerebral tissues and cells compared with the control group. The levels of neuronal necroptosis factors and the co-localization of NeuN and MLKL were also increased in MCAO mice compared with the Sham group. MALAT1 was found to interact with HSP90, and inhibition of HSP90 expression led to decreased phosphorylation levels of neuronal necroptosis factors. Inhibition of MALAT1 expression resulted in decreased co-localization levels of NeuN and MLKL, decreased phosphorylation levels of neuronal necroptosis factors, and reduced necroptosis rate in cerebral tissues. Furthermore, inhibiting MALAT1 expression also led to a shorter half-life of HSP90, increased ubiquitination level, and decreased phosphorylation levels of neuronal necroptosis factors in cells. In conclusion, this study demonstrated that lncRNA MALAT1 promotes neuronal necroptosis in CIR mice by stabilizing HSP90.

Tumor necrosis factor alpha (TNF-a) and glial fibrillary acidic protein (GFAP) are candidate biomarkers for early diagnosis and evaluation of spinal cord injury (SCI). In fact, their concentrations in serum, plasma, or cerebrospinal fluid are highly correlated with the SCI severity and neurological prognosis. Therefore, development of a rapid, ultrasensitive, and accurate technique for dual-detection of these important biomarkers is critical for SCI diagnosis, monitoring, and recovery assessment. We synthesized gold nanorods (AuNRs) with stable aspect ratio and fabricated an AuNR array parallelly aligned on a substrate using evaporation-induced self-assembly. The uniform morphology and neat arrangement of the AuNRs produce many hotspots that increase the sensitivity and reproducibility of the array. For dual detection of TNF-alpha and GFAP, the self-assembled AuNR array substrate was modified with anti-TNF-alpha and anti-GFAP. Then, special Raman probe molecules, specific antibodies, and AuNRs were combined to fabricate surface-enhanced Raman scattering (SERS) tags of AuNR@4-MPBA and AuNR@NBA modified with anti-TNF-alpha and anti-GFAP, respectively. Sandwich immunoassay using standard solutions of TNF alpha and GFAP provided limits of detection of 0.023 and 0.018 pg/mL, respectively. The detector was evaluated on concentrations of TNF- a and GFAP in plasma of SCI rat models. Cross-validation against ELISA (enzyme-linked immunosorbent assay) indicated that the SERS sandwich immunoassay detector based on AuNR array substrate has a high dual-detection accuracy. Overall, the proposed detector provides high sensitivity, reproducibility, stability, and accuracy, and it correctly performs dual detection. The fabricated detector platform seems promising for detection of multiple molecules to achieve early and accurate disease diagnosis in clinical practice.

Small extracellular vesicles (sEVs) secreted by mesenchymal stem cells (MSCs) have shown great therapeutic potential in cerebral ischemia-reperfusion injury (CIRI). In this study, we firstly performed a systematic review to evaluate the efficacy of MSCs-derived sEV for experimental cerebral ischemia/reperfusion injury. 24 studies were identified by searching 8 databases from January 2012 to August 2022. The methodological quality was assessed by using the SYRCLE 's risk of bias tool for animal studies. All the data were analyzed using RevMan 5.3 software. As a result, the score of study quality ranged from 3 to 9 in a total of ten points. Meta-analyses showed that MSCs-derived sEVs could effectively alleviate neurological impairment scores, reduced the volume of cerebral infarction and brain water content, and attenuated neuronal apoptosis. Additionally, the possible mechanisms of MSCs-derived sEVs for attenuating neuronal apoptosis were inhibiting microglia-mediated neuroinflammation. Thus, MSCs-derived sEVs might be regarded as a novel insight for cerebral ischemic stroke. However, further mechanistic studies, therapeutic safety, and clinical trials are required. Systematic review registration. PROSPERO CRD42022312227.

OBJECTIVES:Inflammation especially the overexpression of inflammasome and inflammatory cytokines, is one of the important reasons that affect the occurrence and development of acute cerebral infarction, including the initiation of cerebral infarction, the progress and recovery of post-infarction injury. This study aims to explore expressions of absent in melanoma 2 (AIM2), interleukin-1β (IL-1β), and interleukin-18 (IL-18) in plasma of patients with acute cerebral infarction and its significance. METHODS:A total of 85 patients with acute cerebral infarction were enrolled in the cerebral infarction group. They were assigned into mild, moderate, and severe groups according to the severity of neurological deficits. They were assigned into small, middle, and large cerebral infarction groups according to the area of cerebral infarction. They were assigned into a good prognosis group and a poor prognosis group according to the Modified Rankin Scale (mRS) score on the 90th day after the onset. A total of 85 healthy controls were selected as a control group. The levels of AIM2, IL-1β, and IL-18 in plasma of the cerebral group and the control group were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS:The levels of plasma AIM2, IL-1β, and IL-18 in the cerebral infarction group were significantly higher than those in the control group (all P<0.001). In the cerebral infarction group, the expression levels of plasma AIM2, IL-1β, and IL-18 were as follows: The severe neurological deficitc group>the moderate group>the mild group, the large area of cerebral infarction group>the middle area group>the small area group, and the poor prognosis group> the good prognosis group (all P<0.05). The levels of plasma AIM2 were positively correlated with National Institute of Health Stroke Scale (NIHSS) score, the cerebral infarction area, and the mRS score (r=0.791, r=0.710, r=0.763, respectively, all P<0.001). The levels of plasma IL-1β were positively correlated with the NIHSS score, the cerebral infarction area, and the mRS score (r=0.716, r=0.690, r=0.688, respectively, all P<0.001). The levels of plasma IL-18 were positively correlated with the NIHSS score, the cerebral infarction area, and the mRS score (r=0.714, r=0.638, r=0.653, respectively, all P<0.001). The level of plasma AIM2 was positively correlated with that of IL-1β and IL-18 (r=0.828, r=0.751, both P<0.001). CONCLUSIONS:Expressions of AIM2, IL-1β, and IL-18 are up-regulated in the plasma of patients with acute cerebral infarction, and they are closely related to the severity of neurological deficit, cerebral infarction area, and prognosis in patients with acute cerebral infarction, suggesting that AIM2, IL-1β, and IL-18 may play an important role in the occurrence and development of acute cerebral infarction.

Objective: To investigate the bioinformatics of differentially expressed proteins in plasma in patients with acute cerebral infarction associated with H-type hypertension. Methods: Gene chip public database (gene expression omnibus, GEO) GDS4521 chip data, in the chip in 30 cases of H patients with acute cerebral infarction with high blood pressure and age, gender, matching the 20 H hypertension patients as the research object, collects the mononuclear cells (PBMCs) is used to detect the gene chip, using the GO (gene Ontology, GO), protein function analysis tools such as KEGG, screening and analysis of enrichment of differentially expressed genes function and related signaling pathway. Results: 31 genes in PBMCs were significantly changed in h-type hypertension and H-type hypertension with acute cerebral infarction, 32 of which were increased and 2 of which were decreased. GO analysis showed that in terms of biological processes, the genes related to inflammatory response and neutrophil chemotaxis were the most. In terms of molecular function, chemokine activity-related genes are the most. KEGG signaling pathway analysis showed that the most differentially expressed genes were located in the TNF signaling pathway. Conclusion: H-type hypertension with acute cerebral infarction has a variety of functional proteins and signaling pathways changes, suggesting that inflammatory response in H-type hypertension with acute cerebral infarction recovery period may still play a role in the prognosis and reactivation of the disease.

The pathogenesis of neuromyelitisoptica spectrum disorders (NMOSD) is influenced by a combination of genetic and environmental factors, including infectious agents. Several viral and bacterial infectious diseases are related to the onset and the relapse of NMOSD.The cases of NMOSD with bacterial meningitis have also been reported. Different infections play different role and outcomes in NMOSD.However,the major pathogenesis contains bystander activation, molecular mimicry, and systemic infection triggering independent central nervous system diseases. Viral and bacteria infections are closely related to NMOSD.We recommend that patients with NMOSD and altered mental status need to complete infectious examination, including a lumbar cone puncture.

H-type hypertension, defined as a combination of hypertension and hyperhomocysteinemia (Hhcy), is associated with atherosclerosis and, therefore, increased stroke risk. However, the role of hypertension and Hhcy in high-risk stroke populations has not been studied. The present study investigated the prevalence of H-type hypertension in a high-risk stroke population of Hainan Province, China and to assess possible joint effects between hypertension and Hhcy for increased carotid intima-media thickness (CIMT). In this community-based cross-sectional study, 959 high-risk stroke subjects (age, 65.8 +/- 10.8 years; 46.6% men) were recruited from Hainan Province, China. The demographic and clinical characteristics were collected, and blood samples were obtained. Analysis of variance or chi-square tests were performed to compare variates among groups based on both homocysteine levels and blood pressure status. The associations of hypertension and Hhcy with increased CIMT were evaluated through logistic regression. The prevalence of H-type hypertension was 34.8% in this population, with a higher ratio of H-type hypertension in men than in women. Compared with the normotension and normal homocysteine subgroup, the risk of increased CIMT was significantly higher in the subgroup with hypertension and Hhcy (odds ratio [OR] = 2.639; 95% confidence interval [CI], 1.690-4.091) after adjusting for age and sex. Increased CIMT was affected by an additive synergetic interaction between Hhcy and hypertension (synergy index = 1.105). It emphasized the clinical importance of anti-hypertension and lowering Hhcy in the high-risk stroke population.

目的 探讨血清碱性磷酸酶(ALP)、脂蛋白相关磷脂酶A2(Lp-PLA2)对缺血性脑卒中后认知功能障碍的预测价值及缺血性脑卒中后认知功能障碍发生的影响因素,为临床诊治提供依据.方法 选取2017年1月至2018年12月在中南大学湘雅医学院附属海口医院神经内科诊治的缺血性脑卒中患者203例,依据蒙特利尔认知评估量表(MoCA-B)评分将患者分为认知功能障碍组(88例)和非认知功能障碍组(115例),检测血清ALP、Lp-PLA2水平.采用SPSS 22.0软件进行独立样本t检验、Mann-Whitney U检验和x2检验.应用受试者工作特征(ROC)曲线评价血清ALP、Lp-PLA2对缺血性脑卒中后认知功能障碍的预测价值.采用多因素logistic回归分析缺血性脑卒中后认知功能障碍的影响因素.结果 认知功能障碍组血清ALP、Lp-PLA2水平高于非认知功能障碍组,差异均有统计学意义(P＜0.01).血清ALP、Lp-PLA2预测缺血性脑卒中后认知功能障碍的曲线下面积分别为0.823、0.858,临界值为75.52 U/L和253.31 ng/ml时,灵敏度分别为81.82％、72.73％,特异度分别为82.61％、84.35％.多因素logistic回归分析显示,年龄≥65岁(OR=1.032)、同型半胱氨酸(Hcy)≥ 15.26μmol/L(OR=1.112)、超敏C反应蛋白(hs-CRP)≥4.45 mg/L(OR=2.131)、ALP≥75.52 U/L(OR=1.573)和Lp-PLA2≥253.31 ng/ml(OR=1.021)与缺血性脑卒中后认知功能障碍发生高风险相关,教育年限≥6年(OR=0.820)与缺血性脑卒中后认知功能障碍发生低风险相关,均有统计学意义(P＜0.05,P＜0.01).结论 血清ALP和Lp-PLA2与缺血性脑卒中后认知功能障碍密切相关,可作为预测缺血性脑卒中后认知功能障碍的生物标志物.

Ischemic stroke is a common clinical cardiovascular disease and often accompanied by central nervous system injury. It often causes paralysis or loss of motor function after central nervous system injury and significantly reduces the patient's quality of life. At present, there is no effective treatment strategy for nerve damage caused by ischemic stroke. Therefore, it is urgently need to explore effective treatment targets. The protein expression of SOX5, VEGF and apoptosis related proteins were measured by western blot. The mRNA expression of SOX5 and VEGF were detected by RT-qPCR. The concentration of S100B and GFAP which are related to nerve damage were detected using ELISA assay. The transcriptional regulation of SOX5 on VEGF was detected using ChIP-PCR and dual luciferase reporter gene assays. The cell apoptosis was measured by TUNEL assay and cell viability was detected by CCK-8 assay. In our study, we found that the expression of SOX5 was significantly reduced when LPS induced apoptosis in PC-12 cells. Overexpression of SOX5 repaired LPS-induced apoptosis. SOX5 promotes VEGF expression as a transcription factor to activate the PI3K/AKT pathway. VEGF also repairs nerve injury and brain tissue injury caused by ischemic stroke. In conclusion, SOX5 transcription regulates the expression of VEGF to activate the PI3K/AKT pathway, which repaired nerve damage caused by ischemic stroke. Therefore, SOX5 could be a new targetto regulate VEGF which can repair nerve injury induced by ischemic stroke.

Proteasome subunit α type 6 (PSMA6) -8 C>G polymorphism has been reported to be associated with the susceptibility to ischemic stroke (IS). Given the inconsistent results, we conducted a meta-analysis to assess the association between this SNP and the risk of IS. Articles based on the association between this variant and IS were searched in PubMed, EMBASE, Chinese National Knowledge Infrastructure (CNKI), Wanfang, and Google Scholar database published before August 1, 2016. Pooled odds ratios (OR) and 95% confidence intervals (CI) were calculated using fixed-effect or random-effect models in five genetic models (allele, co-dominant, dominant, recessive, and additive model) according to heterogeneity among studies. In the combined analysis, this variant was found to be associated with a decreased risk of IS under heterozygous and dominant model, while was proven to be associated with a reduced risk of large vessel disease (LVD) under allele, heterozygous and dominant model. Based on ethnic subgroup analysis, in Caucasian population, the risk of ischemic stroke was reduced in each of the genetic models, while the risk of large vessel disease was decreased in the allele, heterozygous and dominant model. As for African-American, lower IS risk of IS was only observed under allele model. Null association existed in Asian population. Our meta-analysis show that PSMA6 -8 C>G polymorphism is more likely to be associated with a decreased risk of IS and LVD in combined analysis, which is also present in Caucasian and African-American population.

Objective? To evaluate the relationship of methy-lenetetrahydrofolate reductase (MTHFR) gene polymorphisms and gene- risk factors interactions with the risk of carotid intima-media thickniss(CIMT) in a Hainan population which without stroke. Method Participants were selected from populations with high risk of stroke ,but without sroke yet.The SNPstats platform was used to investigate the association between MTHFR gene polymorphisms and CMIT under 5 genetic models (dominant, recessive, codominant,over dominant and additive models). Gene-risk factors interactions analysis was then performed by multifactor di-mensionality reduction(MDR).Results Hypertension,diabetes and hyperhomocysteinemia (Hhcy)would accelerate CIMT. The allele frequency of 677C＞T and 1298A＞C has no significant difference(P＞0.05). The 5 genetic models of 677C＞T had no significant correlation with CIMT(P＞0.05).The codominant(A/C,C/C),dominant(A/C-C/C)and Log-additive of 1298A＞ C had significant correlation with CIMT(P ＜ 0.05). Haplotype C-C had significant correlation with CIMT (P=0.0046).No significant multiplicative and additive interactions were observed in terms of CIMT between 1298 A＞C polymorphism and age,gender,hypertension,hyperlipidemia,diabetes,atrial fibrillation(AF),smoking ,family histry of stroke, drinking, BMI,and homocysteine(Hcy). Conclusions MTHFR 1298A ＞ C gene polymorphisms had significant correlation with CIMT in unattact populations with high risk of stroke in Hainan, while 677C＞T gene polymorphisms had nothing. MTHFR gene polymorphisms ,gene- risk factors interactions and CIMT had no significant results.

OBJECTIVE:Inflammasome contributes to ischemic brain injury by inducing pyroptosis and inflammation. The aim of this study is to unravel the mechanism of long non-coding RNA (lncRNA) maternally expressed gene 3 (MEG3)-mediated regulation of absent in melanoma 2 (AIM2) inflammasome during cerebral ischemia/reperfusion (I/R). METHODS:In vivo middle cerebral artery occlusion (MCAO) rat model and in vitro oxygen-glucose deprivation/reperfusion (OGD/R)-treated neurocytes model were generated. TTC, H&E staining and TUNEL were performed to assess the cerebral ischemic injury. LDH and MTT assays were used to detect cell viability and cytotoxicity. qRT-PCR was used to detect the expression levels of MEG3, miR-485 and AIM2. Immunohistochemistry (IHC) and immunofluorescence were conducted to detect the AIM2 expression. ELISA and Western blotting were performed to determine the secretion and protein levels of inflammasome signaling proteins. Dual luciferase reporter assay and Ago2-RIP were used to validate the direct interaction among MEG3, miR-485 and AIM2. RESULTS:In both MCAO rats and OGD/R-treated neurocytes, MEG3 and AIM2 were significantly up-regulated, whereas miR-485 was down-regulated. MCAO induces pyroptosis and release of IL-1β and IL-18 in ischemia brain. MEG3 acted as a molecular sponge to suppress miR-485, and AIM2 was identified as a direct target of miR-485. Knockdown of MEG3 inhibited OGD/R-induced pyroptosis and inflammation, and lack of MEG3 inhibited caspase1 signaling and decreased the expression of AIM2, ASC, cleaved-caspase1 and GSDMD-N. While overexpression of MEG3 exerted opposite effects. CONCLUSION:MEG3/miR-485/AIM2 axis contributes to pyroptosis via activating caspase1 signaling during cerebral I/R, suggesting that this axis may be a potent therapeutic target in ischemic stroke.