目的 探究急性后循环供血区缺血性脑梗死患者血管再通术后24 h内Alberta卒中操作早期CT评分(Alberta stroke program early CT score,ASPECTS)联合血清血红素氧合酶-1(Heme oxygenase-1,HO-1)、血管内皮生长因子(Vascular endothelial growth factor,VEGF)水平对患者短期不良预后的预测价值.方法 选取医院2019年1月-2020年1月收治的62例急性后循环供血区缺血性脑梗死患者,均行血管再通术;术后随访3个月,根据改良Rankin量表评分(Modified Rankin scale,mRS)进行神经功能评定,分为预后良好组、预后不良组,采用单因素分析比较2组术后24h内ASPECTS评分、血清HO-1,VEGF水平等可能影响患者短期不良预后的因素;采用Logistic回归分析急性后循环供血区缺血性脑梗死患者血管再通术后影响患者短期不良预后的危险因素,并绘制受试者工作特征(Receiver operating characteristic,ROC)曲线评估术后ASPECTS评分联合血清HO-1,VEGF水平对患者短期不良预后的预测价值.结果 根据随访3个月后mRS评分判定,62例患者中40例归为预后良好组,22例归为预后不良组.单因素分析中2组患者性别、年龄、体质量指数、是否吸烟、酗酒,高脂血症、糖尿病、冠心病、既往脑卒中、高血压病等既往病史、治疗前血压、术后24 h内血压、发病至入院时间、术后24 h内实验室检查指标(白细胞计数、血小板、血糖、血尿素、血肌酐、胱抑素C)水平均无明显差异(P＞0.05);预后良好组既往病史中心房颤动占比(12.50％)低于预后不良组(45.45％)(P＜0.05),术后24 h内NIHSS评分低于预后不良组(P＜0.05),术后24 h内ASPECTS评分高于预后不良组(P＜0.05).术后24 h内实验室检查指标中预后良好组血清HO-1水平高于预后不良组,VEGF水平低于预后不良组(P＜0.05).经Logistic回归分析显示,心房颤动史、术后24 h内NIHSS评分、术后24 h内ASPECTS评分、血清HO-1,VEGF水平是影响急性后循环供血区缺血性脑梗死患者血管再通术后短期不良预后的危险因素.ROC曲线分析显示,术后24 h内ASPECTS评分联合血清HO-1,VEGF水平预测急性后循环供血区缺血性脑梗死患者血管再通术后短期不良预后的敏感度、准确度、AUC分别为93.67％、84.36％、0.873,均高于血清HO-1水平、VEGF水平、ASPECTS评分单独预测.结论 术后24 h内ASPECTS评分、血清HO-1,VEGF水平与急性后循环供血区缺血性脑梗死患者血管再通术后短期不良预后密切相关,三者联合预测的准确性更好.

Objective:To investigate the efficacy of different preferred thrombectomy strategies for embolic acute vertebrobasilar artery occlusion (AVBAO).Methods:Forty-four patients with embolic AVBAO who underwent endovascular treatment in Department of Neurology, Nanyang Central Hospital from January 2019 to June 2021 were included in the study. Patients were divided into stent-retriever thrombectomy group ( n=27) and aspiration thrombectomy group ( n=17) according to different preferred thrombectomy strategies. Modified Rankin scale (mRS) was used to evaluate the prognoses of these patients 90 d after surgery; the differences of clinical data, surgery-related characteristics, prognoses and complications between the two groups were compared. Results:There was no significant difference between the 2 groups in terms of time from onset to puncture, sites of target vessel occlusion, proportion of patients accepted intraoperative remedial measures, and successful recirculation rate of target vessels ( P>0.05). Compared with the aspiration thrombectomy group, the stent-retriever thrombectomy group had significantly decreased utilization rate of middle catheters, significantly increased retrieval attempts in thrombectomy, statistically lower re-recanalization rate of first-time thrombectomy on the target vessels, significantly longer time from puncture to re-recanalization, and significantly higher incidence of new embolism ( P<0.05). There was no significant difference between the 2 groups in incidences of vascular rupture and postoperative spontaneous intracerebral hemorrhage (sICH), and good prognosis rate 90 d after surgery ( P>0.05). Conclusion:For embolic AVBAO patients, similar recanalization and short-term good prognosis can be obtained by aspiration thrombectomy to those by stent-retriever thrombectomy; besides that, aspiration thrombectomy has advantages as shorter recanalization time, less new embolic complications and higher re-recanalization rate of first-time thrombectomy.

目的 探究高压氧联合回神醒脑汤改善脑梗死患者神经、认知和肢体功能及生活能力的作用分析.方法 选取2018年3月-2019年10月在医院接受治疗的86例脑梗死患者为研究对象,采用随机数字表法分为对照组与观察组,每组43例.对照组采用常规治疗,观察组在此基础之上采用高压氧联合回神醒脑汤治疗,疗程4周.观察两组临床疗效、神经功能、认知功能、肢体运动功能、日常生活能力及血清学指标.结果 观察组总有效率高于对照组(P＜0.05).治疗后,两组C反应蛋白(CRP)、D-二聚体(D-Dimer)、血栓调节蛋白(TM)、白介素-6(IL-6)和脑源性神经营养因子(BDNF)较治疗前降低(P＜0.05),且观察组低于对照组(P＜0.05).治疗后,两组NIHSS和MMSE评分较治疗前降低(P＜0.05),且观察组低于对照组(P＜0.05);治疗后,两组FMA和ADL评分较治疗前升高(P＜0.05),且观察组高于对照组(P＜0.05).结论 高压氧联合回神醒脑汤能够有效改善患者的神经和肢体运动功能障碍,并能显著提高患者的认知功能和日常行为能力,疗效确切.

目的:探究脑苷肌肽辅助治疗脑梗死后认知功能障碍的临床效果.方法:选取2018年9月～2020年5月接受治疗的130例脑梗死后认知功能障碍患者为受试对象,根据随机数字表法分为观察组与对照组,各65例.对照组予以神经内科基础治疗,观察组在对照组基础上联合脑苷肌肽辅助治疗.比较两组治疗前后神经功能(神经功能缺损评分、精神状态评分)、促炎因子(血小板活化因子、亲环素A、单核细胞趋化蛋白-1)、血清神经生化标志物(中枢神经特异蛋白、人脑髓鞘碱性蛋白、神经元特异性烯醇化酶)变化情况,并记录治疗过程中的不良反应,评估用药安全.结果:治疗14d后,两组患者神经功能缺损评分、血小板活化因子、亲环素A、单核细胞趋化蛋白-1、中枢神经特异蛋白、人脑髓鞘碱性蛋白、神经元特异性烯醇化酶水平较治疗前降低,且观察组低于对照组(P＜0.05);两组患者精神状态评分较治疗前升高,且观察组高于对照组(P＜0.05);观察组患者畏寒、低热、头晕等不良反应发生率均高于对照组(P＜0.05),而两组皮疹、白细胞减少不良反应发生率比较,差异无统计学意义(P＞0.05).结论:脑苷肌肽辅助治疗可明显改善脑梗死患者认知功能障碍,有利于神经功能的恢复,且还可显著降低炎症介质水平,但其不良反应较多,需引起重视,临床可根据实际情况进行选择.

目的:基于神经调节蛋白(NRG)/表皮生长因子受体(ErbB)信号通路探讨三七总皂苷(TSPN)对偏头痛大鼠的保护机制.方法:将SD大鼠随机分为对照组、模型组、TSPN(10 mg/kg)组和TSPN(10 mg/kg)+NRG/ErbB信号通路抑制剂AG825(50μg/kg)组(TSPN+抑制剂组),每组12只.给药1周后,模型组、TSPN组和TSPN+抑制剂组大鼠用2 mL硝酸甘油溶液(剂量为10 mg/kg)皮下注射建立偏头痛模型,对照组相同部位注射等体积的生理盐水.各组进行行为学评分;苏木精-伊红(HE)染色检测脑组织神经元形态;酶联免疫吸附分析(ELISA)检测血清中一氧化氮(NO)、白细胞介素6(IL-6)和IL-1β水平,以及脑组织中5-羟色胺(5-HT)、β-内啡肽(β-EP)和降钙素基因相关肽(CGRP)含量;Western blot法检测脑组织中NRG1、ErbB2、ErbB3、ErbB4、caspase-3和cleaved caspase-3蛋白水平.结果:模型组神经元空泡状明显,细胞萎缩、坏死严重,出现局部坏死、血管阻塞现象;TSPN组神经元得到一定修复,仅存在部分空泡现象;TSPN+抑制剂组细胞空泡现象明显,出现部分血管阻塞现象.与对照组相比,模型组在0～30 min、60～90 min和120～150 min中行为学评分,血清中NO、IL-6和IL-1β水平,脑组织中CGRP含量,以及脑组织中cleaved caspase-3/caspase-3蛋白水平显著升高(P<0.05),脑组织中5-HT和β-EP含量及NRG1、ErbB2、ErbB3和ErbB4蛋白水平显著降低(P<0.05);与模型组相比,TSPN组上述指标有所缓解,耳红出现时间延长(P<0.05),耳红消失时间缩短(P<0.05);与TSPN组相比,NRG/ErbB信号通路抑制剂AG825逆转TSPN对上述指标的缓解.结论:TSPN可能通过激活NRG/ErbB信号通路实现对偏头痛的缓解.

目的 研究纳洛酮联合依达拉奉对急性脑出血患者认知功能的影响.方法 选取2019年4月至2020年4月南阳市中心医院收治的200例急性脑出血患者作为研究对象,根据治疗方式的不同分为对照组和观察组,每组100例.对照组采用手术治疗,观察组采用纳洛酮联合依达拉奉治疗.比较两组治疗后临床疗效、认知功能及血浆纤维蛋白原(FIB)、D-二聚体(D-D)水平.结果 观察组治疗后临床疗效高于对照组(P＜0.05).两组治疗前NIHSS评分、Fugl-Meyer评分比较,差异无统计学意义(P＞0.05).观察组治疗后NIHSS评分、Fugl-Meyer评分优于对照组(P＜0.05).两组治疗前血浆FIB、D-D水平比较,差异无统计学意义(P＞0.05).观察组治疗后FIB、D-D水平优于对照组(p＜0.05).结论 纳洛酮联合依迭拉奉可提高急性脑出血患者疗效,改善认知功能,降低炎症反应.

目的:分析动脉粥样硬化性血栓性脑梗死患者中,应用奥扎格雷钠联合阿托伐他汀钙的临床效果.方法:选取在我院接受治疗的150例动脉粥样硬化性血栓性脑梗死患者,收治的时间为2018-09～2020-06.按照不同的治疗方法,将所有的患者分为研究组和常规组,各75例.将两组治疗后的血液流变学指标以及脑血管生理功能变化情况进行对比.结果:治疗后,相比于常规组,研究组的全血黏度高切、全血黏度低切、血液黏度明显低(P<0.05);相比于常规组,研究组的A峰、EF、LVIDs、MVCF、SV水平明显高(P<0.05);相比于常规组,研究组的E峰值明显低(P<0.05);两组的不良反应发生率相比不具有差异性(P>0.05).结论:奥扎格雷钠联合阿托伐他汀钙片在动脉粥样硬化性血栓性脑梗死患者中的应用效果较好,可以快速改善患者的血液状态,加速血液的流动性,减少血小板的聚集,降低血液黏稠度,从而利于患者身体的恢复.

目的 分析前列地尔联合尼莫地平对动脉瘤性蛛网膜下腔出血(aSAH)后脑血管痉挛患者炎症因子水平的影响.方法 将143例患者分为对照组71例和观察组72例.对照组给予前列地尔进行治疗,观察组给予前列地尔加尼莫地平进行治疗.比较2组白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、超敏C反应蛋白(hs-CRP)和肿瘤坏死因子(TNF-ot)水平、治疗效果、不良反应、认知功能.结果 治疗后,观察组炎症因子水平、不良反应总发生率均低于对照组,总有效率、认知功能评定测验(LOTCA)评分均高于对照组,差异有统计学意义(P＜0.05).结论 前列地尔联合尼莫地平治疗aSAH后脑血管痉挛患者安全有效,可有效降低患者机体炎症因子水平和不良反应发生率,改善患者认知功能.

背景:课题组前期研究发现,骨髓间充质干细胞来源外泌体可减少脊髓损伤区域反应性星形胶质细胞的数量,有利于损伤后运动功能的改善.目的:探索骨髓间充质干细胞外泌体对脊髓损伤后A1型星形胶质细胞活化的影响及其在损伤后修复中的作用.方法:培养SD大鼠骨髓间充质干细胞,利用超速离心法提取细胞上清液中的外泌体并进行鉴定.将80只成年雄性SD大鼠随机分为假手术组、脊髓损伤模型组、骨髓间充质干细胞组、骨髓间充质干细胞外泌体组,每组20只.除假手术组外,其余3组均使用Allen法制备脊髓损伤模型,术后2 h通过尾静脉注射等体积的PBS、骨髓间充质干细胞悬液、骨髓间充质干细胞外泌体悬液,每隔2 d注射1次,共注射3次.脊髓损伤术前、术后1,3,7,14,28 d采用BBB评分法评估后肢运动功能;术后7 d通过苏木精-伊红染色、透射电镜观察脊髓组织形态,免疫荧光检测A1型星形胶质细胞数量、NeuN阳性神经元数量、caspase-3阳性凋亡细胞数量,ELISA法检测脊髓组织肿瘤坏死因子α、白细胞介素1α水平,Western blot检测脊髓组织C3、GFAP的表达水平.结果与结论:①骨髓间充质干细胞外泌体在透射电镜下呈茶托状,表达外泌体标志CD63、CD9;②骨髓间充质干细胞外泌体减少脊髓组织炎症细胞浸润、减轻髓鞘脱失、轴突退变,降低炎症因子肿瘤坏死因子 α、白细胞介素1α水平和C3的表达水平,降低A1型星形胶质细胞数量,减少细胞凋亡,提高残余神经元数量,改善肢体运动功能;③以上结果提示,骨髓间充质干细胞外泌体能减少脊髓A1型星形胶质细胞的活化,促进脊髓损伤后运动功能恢复.

AIM:To study the influence of bone marrow mesenchymol stem cell-drived exosomes(BMSC-exo-somes)on hindlimb activity,and the numbers of reactive astrocytes and residual neurons in spinal cord injury(SCI)rats. METHODS:BMSCs were cultured using the whole bone marrow adherent culture method and surface markers CD 90 and CD34 were verified by flow cytometry.Exosomes were isolated by ultracentrifugation and the morphology of exosomes was observed under transmission electron microscope.The protein markers CD63 and CD9 were verified by Western blot.After exosomes were applied to SCI rats,the Basso,Beattie and Bresnahan locomotor rating scale score,the Nissl staining of the lesion site,and the numbers of reactive astrocytes and residual neurons were assessed at various time points.RESULTS:Transmission electron microscopic observation revealed the presence of saucer -shaped vesicles.BMSC-exosomes were found to express high levels of CD63 and CD9.Compared with injury group,significant improvement of hindlimb activity scores from day 14 after injury in treatment group was observed(P<0.05),and less reactive astrocytes and more residual neu-rons from day 7 after injury were also observed(P<0.05).CONCLUSION:BMSC-exosomes inhibit reactive astrocytes and death of neurons,and improve hindlimb activity in the rats after SCI.

我们通过研究脊髓损伤(SCI)后损伤节段A1型星形胶质细胞(A1-Ast)数量随时间的变化规律,寻找特异性抑制A1-Ast活化的最适时间点。现报道如下。一、材料与方法1.材料:SD大鼠;小鼠抗大鼠胶质纤维酸性蛋白(GFAP)单克隆抗体;兔抗大鼠C3单克隆抗体,山羊抗小鼠二抗,山羊抗兔二抗;IH-0400脊髓打击器;2.SCI模型制备:10%水合氯醛麻醉,背正中切口,暴露、去

BACKGROUND/AIMS:Neurotoxic A1 astrocytes are induced by inflammation after spinal cord injury (SCI), and the inflammation-related Nuclear Factor Kappa B (NFκB) pathway may be related to A1-astrocyte activation. Mesenchymal stem cell (MSC) transplantation is a promising therapy for SCI, where transplanted MSCs exhibit anti-inflammatory effects by downregulating proinflammatory factors, such as Tumor Necrosis Factor (TNF)-α and NFκB. MSC-exosomes (MSC-exo) reportedly mimic the beneficial effects of MSCs. Therefore, in this study, we investigated whether MSCs and MSC-exo exert inhibitory effects on A1 astrocytes and are beneficial for recovery after SCI.METHODS:The effects of MSC and MSC-exo on SCIinduced A1 astrocytes, and the potential mechanisms were investigated in vitro and in vivo using immunofluorescence and western blot. In addition, we assessed the histopathology, levels of proinflammatory cytokines and locomotor function to verify the effects of MSC and MSC-exo on SCI rats.RESULTS:MSC or MSC-exo co-culture reduced the proportion of SCIinduced A1 astrocytes. Intravenously-injected MSC or MSC-exo after SCI significantly reduced the proportion of A1 astrocytes, the percentage of p65 positive nuclei in astrocytes, and the percentage of TUNEL-positive cells in the ventral horn. Additionally, we observed decreased lesion area and expression of TNFα, Interleukin (IL)-1α and IL-1β, elevated expression of Myelin Basic Protein (MBP), Synaptophysin (Syn) and Neuronal Nuclei (NeuN), and improved Basso, Beattie & Bresnahan (BBB) scores and inclined-plane-test angle. In vitro assay showed that MSC and MSC-exo reduced SCI-induced A1 astrocytes, probably via inhibiting the nuclear translocation of the NFκB p65.CONCLUSION:MSC and MSC-exo reduce SCI-induced A1 astrocytes, probably via inhibiting nuclear translocation of NFκB p65, and exert antiinflammatory and neuroprotective effects following SCI, with the therapeutic effect of MSCexo comparable with that of MSCs when applied intravenously.

Objectives:To investigate whether the exosomes derived from bone marrow stem cells(BMSCs) by intravenous infusion can promote the regeneration of injured spinal cord.Methods:BMSCs from adult rats were enriched and expanded by whole bone marrow culture in vitro,collecting the cell supernatant of P2 generation,by using the ExoQuick Precipitation for separation and purification of exosomes,the exosome markers,CD9,CD63 were analyzed by using Western blot.The MSCs-Exosomes morphology was assessed by transmission electron microscopy(TEM).The rats were induced by spinal cord strike machine to establish the spinal cord models.Exosomes of 500μl(final concentrate of 2001μg/ml) were infused into tail vein.The rats were randomly divided into the following three groups(n=10/group):sham group,control group(SCI+PBS) and exosomes group (SCI+exosomes).Neurological functions were evaluated by the Basso-Beattie-Bresnahan (BBB) score and the slope plate experiment score at 1,3,7,14,21,28 days after modeling.Histomorphological changes of spinal cord were observed at 28 days after modeling by hematoxylin-eosin(HE) staining and luxol fast blue (LFB) staining,Survivor number of nerve cells was observed at 28 days after modeling by Nissl staining.Results:TEM observation of MSCs-Exosomes revealed the presence of spherical vesicles with a typical cup-shape.The size distribution profile displayed a homogeneous population from 40-100nm,which was consistent with the literature description,MSCs-Exosomes were found to express high levels of CD9,CD63 by using Western blot.After modeling,BBB score and sloping plate experiment score in each time point were normal in sham group,but in control group and exosomes group were significantly lower than those in sham group(P＜0.05),the scores of BBB in exosomes group were 6.30±0.95,12.70±1.57,16.60±1.08,17.00±0.67 at 7,14,21,28 days respectively after modeling,which were higher than those in control group of 2.50±1.08,6.90±0.99,10.50±0.85,12.50±1.08(P＜0.05) respectively,the scores of slope plate experiment in exosomes group were 43.00±3.50,55.50±4.38,62.50±2.64,65.00±3.33 respectively at 7,14,21,28 days after modeling,which were higher than those in control group of 34.00±3.16,43.00±4.22,49.00±4.59,52.50±4.25 (P＜0.05) respectively.HE,LFB and Nissl staining showed nerve cells with normal morphology in sham group at 28 days after modeling,in control group,spinal cord porosis,disorder of myelin sheath and necrosis of nerve cells were found.The injured spinal cord was restored better and the number of neuronal survival was better in exosomes group than that in control group.TEM observation of MSCs-Exosomes revealed the presence of spherical vesicles,with a typical three-dimensional circular or cup-shape.The size distribution profile displayed a homogeneous population from 40-100nm with a complete lipid membrane structure in the peripheral region with low density content.Conclusions:Intravenous administration of MSCs-Exosomes promotes the restoration of injured spinal cord and improves motor functions,which plays a protective role in the repair of spinal cord injury.

Par6α (partitioning defective 6 homologue alpha), a component of the Par3/Par6/aPKC complex, was recently shown to be essential for axon specification during neuronal development. However, the biological functions and regulatory mechanisms of Par6α in the mesenchymal stem cell (MSC) differentiation process have not been investigated. In this study, we found that the expression of let-7f-5p was downregulated during differentiation of bone marrow-derived MSCs to neuron-like cells. Interestingly, Par6α was predicted to be a target gene of let-7f-5p by computerized analysis and the luciferase reporter assay. Using gain- and loss-of-function approaches, we found that expression of Par6α was inversely correlated with let-7f-5p levels during differentiation (p < 0.05). By silencing Par6α using siRNAs, we demonstrated that Par6α was necessary for MSC neuronal differentiation. Altogether, our studies proved that inhibition of let-7f-5p facilitates induction of MSCs into neuron-like cells by directly targeting Par6α.