一氧化氮合酶与脑缺血早期神经损伤关系研究

目的:探讨大鼠脑缺血早期脑内3种亚型一氧化氮合酶(nitric oxide synthase,NOS)的表达及作用。方法:采用免疫组织化学方法,用三型NOS的多克隆抗体检测大鼠局灶性脑缺血15、30min及1、2、3、6 h NOS在脑内的表达情况。结果:局灶性脑缺血1 h内缺血侧基底节区内神经元型NOS(nNOS)表达上调;脑缺血1 h内在缺血区小血管壁及3 h时在缺血边缘区小血管壁出现内皮型NOS(eNOS)两次上调。局灶性脑缺血6 h内神经组织中未见诱生型NOS(iNOS)表达。结论:在脑缺血的超早期出现结构型NOS(nNOS及eNOS)的上调表达,此时可能同时存在nNOS的神经损伤作用及eNOS的神经保护作用。     

一氧化氮合酶在脑缺血再灌注中的双重作用

目的 探讨短暂脑缺血再灌注后大鼠脑内3型一氧化氮合酶(nitric oxide synthase，NOS)的表达及作用，为脑缺血治疗提供理论依据。方法 采用免疫组织化学方法，用3型NOS的多克隆抗体检测大鼠局灶性脑缺血2h再灌注15min及22h NOS在脑内的表达情况。结果 大鼠脑缺血2h再灌注15min，在脑缺血边缘区的血管壁及神经细胞出现内皮型一氧化氮合酶(endothelial nitric oxide synthase，eNOS)上调表达；脑缺血2h再灌注22h，在脑梗死区内表达神经元型一氧化氮合酶(neuronal mitric oxide synthase，nNOS)的神经细胞减少，并出现表达诱导型一氧化氮合酶(inducible nitric oxide synthase，iNOS)的胶质细胞，同时梗死边缘区血管及神经细胞出现eNOS及iNOS的上调表达。结论 在短暂脑缺血再灌注早期，缺血区周围可能有eNOS相关的保护机制；亚急性期eNOS及iNOS的保护及损伤机制并存；因此，在短暂脑缺血早期恢复灌注后予选择性iNOS抑制剂及促进eNOS活性有可能减少迟发性神经损伤。     

重症肌无力患者IgG对大鼠脑内NOS不同时间表达的影响

目的观察重症肌无力(myasthenia gravis,MG)患者IgG(AchRab)对大鼠脑内一氧化氮合酶(NOS)表达的影响,探讨NOS在MG中造成中枢神经系统损害的机制.方法将AchRab IgG或健康人的IgG注入大鼠侧脑室,1次/d,连续4次.免疫组化方法观察不同时间点大鼠脑皮质、海马及杏仁核神经元型一氧化氮合酶(nNOS)和诱导型一氧化氮合酶(iNOS)的表达变化.结果侧脑室注射后1周实验组大鼠皮质、海马神经元nNOS表达量明显减少,后2周实验组皮质、海马神经元nNOS表达下降更为明显,同时杏仁核神经元nNOS表达量也减少;实验组及对照组脑内细胞均未见iNOS表达.结论AchRab侧脑室内注射可引起大鼠皮质、海马及杏仁核神经元nNOS表达量减少,且2周内这种减少效应随时间延长而增强,但未能诱导脑内细胞iNOS表达,提示AchRab尚可通过抑制大鼠中枢神经系统nNOS表达,降低脑内正常的一氧化氮浓度,减弱一氧化氮对脑组织的保护作用,增加神经元的易损性.     

颅脑损伤后局部组织一氧化氮合酶表达的实验研究

目的研究一氧化氮合酶(nitricoxidesynthase,NOS)基因在实验性大鼠颅脑创伤后局部组织中表达.方法应用免疫组化技术和高清晰度彩色病理图像分析系统,对大鼠脑组织神经细胞中诱导型一氧化氮合酶(iNOS)的表达进行了检测.结果颅脑创伤后大鼠局部及周围神经细胞中有NOS阳性产物表达并具时程特点,伤后2h平均积分光密度(ODI)较0.5h升高不显著(P0.05),伤后6h、12h、24hODI较0.5h升高非常显著(P<0.01).结论在急性脑损伤后的病理过程中,脑组织中诱导型一氧化氮合酶被大量合成,可能是造成机体一氧化氮(Nitricoxide,NO)升高的直接原因.     

慢性间断性缺氧诱导一氧化氮合酶表达的研究

目的:建立大鼠缺氧模型,检测神经元型一氧化氮合酶(nNOS)及诱导型一氧化氮合酶(iNOS)的表达情况。方法:1.建立缺氧模型:将SD大鼠置于常压低氧舱中,充入氮气调节氧浓度至所需氧浓度。2.动物分组:(1)急性缺氧组:在低氧舱中缺氧1.5小时。(2)慢性间断性缺氧组:每日在低氧舱中6小时。每周缺氧6天,共缺氧28天。3.采用免疫组化法检测nNOS和iNOS的表达。4.统计学分析检验。结果:急性缺氧后,iNOS、nNOS阳性神经元增加;慢性缺氧后,iNOS、nNOS阳性神经元仍持续增多,慢性缺氧时增加iNOS-IR细胞远远多于nNOS-IR细胞。结论:我们的研究表明缺氧可引起iNOS、nNOS阳性神经元增加,NOS亚型表达时间的不同说明其脑损伤具有阶段性。     

NOS、p38MAPK、Caspase-3介导大鼠脑缺血神经细胞凋亡可能通路的实验研究

目的探讨脑缺血后细胞凋亡发生的可能机制以及神经元型一氧化氮合酶(neuronal nitric oxide synthase,nNOS)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、p38丝裂原活化蛋白激酶(mitogen activated proteinkinasep38,p38MAPK)和半光氨酸蛋白酶-3(caspase-3)在脑缺血后神经细胞凋亡中的共同作用机制。方法采用线栓法闭塞大鼠大脑中动脉(middle cerebral artery occlusion,MACO)建立脑缺血SD大鼠模型,应用透射电镜观察脑缺血对脑组织超微结构的影响,流式细胞仪方法(FCM)分别定量检测细胞凋亡率,半定量RT-PCR检测nNOS、iNOS,p38MAPK和Caspase-3mRNA表达水平。结果透视电镜下脑缺血6h出现核固缩,缺血12h出现细胞核分裂,缺血24h出现凋亡小体;FCM检测细胞凋亡百分率随着缺血时间延长而增加,缺血72h达到高峰,约70.37%;RT-PCR产物的琼脂糖凝胶电泳显示nNOS、iNOS、p38MAPK和Caspase-3mRNA的特异性片段大小分别为501、342、250和342bp,但mRNA表达量不一致,nNOS mRNA主要在缺血早期表达,iNOS、p38MAPK和Caspase-3mRNA在缺血中晚期表达,并在缺血3~5d,后三种基因的表达量达到高峰。结论脑缺血区域发生典型的神经细胞凋亡现象,nNOS来源的NOS在缺血早期发挥神经毒性作用,iNOS来源的NOS在缺血晚期发挥神经毒性作用;NOS,p38MAPK和Caspase-3三种基因的相互关系可能构成介导缺血神经细胞凋亡的通路之一。     

脑缺血后脑组织NOS基因表达的变化与巴曲酶的影响

近年来发现脑缺血后脑组织一氧化氮(NO)含量增高,NO能显著影响缺血后脑水肿及脑梗塞灶的大小,但一氧化碳合成酶(NOS)拮抗剂对脑缺血的影响尚有争论。巴曲酶(batroxobin)为蛇毒酶制剂,我们用巴曲酶治疗急性缺血性脑血管病取得了较好疗效,动物实验也表明巴曲酶可减轻沙土鼠脑缺血后脑水肿程度和卒中症候,但巴曲酶治疗脑缺血是否能直接或间接影响缺血脑组织的NOS基因表达尚未见报道。     

人工合成E-选择素对大鼠局灶性脑缺血再灌注损伤后脑组织一氧化氮合酶表达的影响

目的 探讨人工合成E-选择素对大鼠局灶性脑缺血/再灌注（I/R）损伤后脑组织一氧化氮合酶（NOS）及血清一氧化氮（NO）含量的影响.方法 采用改良的Zea Longa法建立脑I/R损伤模型.66只雄性SD大鼠随机分为对照组、模型组和人工合成E-选择素治疗组（治疗组）.治疗组大鼠采用股静脉注射人工合成E-选择素10 mg·kg-1.应用硝酸盐还原法测定血清中NO含量和免疫组化法检测缺血区脑组织神经型一氧化氮合酶（nNOS）、诱导型一氧化氮合酶（iNOS）阳性细胞数.结果 ①NO：以对照组NO含量为正常生理数据,模型组脑缺血2h/再灌注2～24h NO含量呈上升趋势,24 h时达高峰,72 h有所降低但仍高于对照组,各时间点与对照组比较明显增高（P＜0.01）;治疗组NO变化趋势同模型组,NO含量较模型组减少（P＜0.05）,较对照组增多（P＜0.01）.②NOS：以对照组nNOS、iNOS阳性细胞数为正常生理数据,模型组nNOS阳性细胞在脑缺血2h/再灌注2h后开始表达,12h达高峰,至24h开始降低,各时间点与对照组比较明显增高（P＜0.01）;模型组iNOS阳性细胞在脑缺血2h/再灌注2h开始出现,并持续增多,随时间延长呈上升趋势,24h达高峰,至72 h出现下降,各时间点与对照组比较明显增多（P＜0.01）;治疗组各时间点nNOS、iNOS阳性细胞变化趋势同模型组,但较模型组减少（P＜0.05）,较对照组增多（P＜0.01）.结论 大鼠脑I/R损伤后脑组织NOS活性表达增多,NO浓度升高导致脑组织损伤;人工合成E-选择素通过降低NOS表达,减少NO释放、减轻炎症反应和脑I/R损伤,起脑保护作用.     

神经元型一氧化氮合酶的神经生物学研究进展

神经元型一氧化氮合酶(neuronal nitric oxide synthase,nNOS)主要表达于神经元,在星形胶质细胞和神经干细胞中也有一定水平的表达.不同的mRNA拼接形式产生了nNOS蛋白的5种亚型,包括nNOS-α、nNOS-β、nNOS-μ、nNOS-γ和nNOS-2.nNOS单体不具催化活性,二聚体是...     

依达拉奉对脑缺血-再灌注大鼠的神经保护作用及其机制研究

目的探讨依达拉奉对局灶性脑缺血-再灌注损伤的影响及其可能的神经保护机制。方法用线栓法制备大鼠局灶性脑缺血-再灌注损伤模型,用试剂盒检测脑组织丙二醛(MDA)含量和一氧化氮合酶(NOS)活性;分别测定脑组织含水量;免疫组织化学染色检测水通道蛋白4(AQP-4)的表达水平。结果缺血再灌注损伤后,大鼠脑组织脂质过氧化产物丙二醛含量和一氧化氮合酶活性增高,MDA含量和NOS合酶活性于脑缺血-再灌注后30min开始增高,于3d达到高峰,于7d基本恢复正常;脑组织含水量于脑缺血再灌注后1d开始增高,于3d达到高峰,于7d时下降至大致正常水平;AQP-4的表达于脑缺血-再灌注后6h开始增高,于3d达到高峰,于7d时下降至大致正常水平。依达拉奉干预能显著降低MDA含量和NOS活性,降低脑组织的含水量,并使AQP-4的表达明显下降。结论在脑缺血-再灌注损伤后,依达拉奉能通过清除自由基而减轻脂质过氧化损伤,并通过抑制AQP-4的表达减轻脑水肿,从而起到神经保护作用。     

Factors associated with satisfaction with care among patients with epilepsy

OBJECTIVE: The purpose of the work described here was to determine those variables associated with satisfaction with care among patients with epilepsy. METHODS: We interviewed patients followed at a tertiary epilepsy center. Predictor variables included age, gender, race, education, income, insurance, seizure frequency, and Quality of Life in Epilepsy-10 inventory (QOLIE-10) results. Target variables were the subscales of the Short Form Patient Satisfaction Questionnaire (PSQ-18). We used univariate analysis to identify those variables significantly associated with the subscales and multiple linear regression to determine those independently significant. RESULTS: The study population comprised 193 patients. Lower education and better QOLIE-10 scores were independently associated with general satisfaction with care. The mental health scale was associated with general satisfaction with care. Lower educational level was the only variable independently associated with patient satisfaction with communication, the financial aspect of care, and time spent with physician. CONCLUSION: Lower educational level and better quality of life are the main variables associated with higher general satisfaction with care among patients with epilepsy.     

Treatment with rituximab in patients with polyneuropathy with anti-MAG antibodies

Journal of Neurology -     

Surgical Outcome in Patients with Epilepsy with Occult Vascular Malformations Treated with Lesionectomy

Summary: Purpose: This retrospective study reports the long-term surgical outcome of patients with medically refractory epilepsy and vascular malformations who were treated with lesionectomy. A detailed analysis of surgical failures had been performed in an attempt to define predictors of surgical success and failure.
Methods: Fifteen patients with medically intractable epilepsy and angiographically occult vascular malformations (AOVMs) were treated surgically with lesionectomy at Duke University Medical Center. Lesionectomy consisted of removal of the AOVM and surrounding hemosiderin-stained brain only, without the use of electrocorticography (ECoG) to guide resection.
Results: Eleven (73%) patients are seizure free after lesionectomy. Three showed no significant improvement, and one patient died, presumably after a seizure. Age of onset, duration of seizures, age at resection, and gender did not affect outcome. All patients with neocortical AOVMs in whom EEG findings correlated with the site of the lesion were seizure free after lesional resection. Treatment failures were associated with the presence of multiple intracranial lesions, poorly localized or diffuse EEG findings, discordant positron emission tomography (PET) imaging, or with a lesion in close proximity to the limbic system.
Conclusions: Lesionectomy, with removal of surrounding hemosiderin-stained brain, can be considered the procedure of choice in carefully selected patients with epilepsy with occult vascular malformations.     

Children with ADHD and motor dysfunction compared with children with ADHD only

The purpose of this study was to identify group differences in children with attention-deficit-hyperactivity disorder and motor dysfunction (ADHD-MD) and ADHD only, and to evaluate the medication responsiveness of ADHD-MD. Sixty-three children (49 males and 14 females; mean age 9 years 10 months, SD 2 years 10 months) underwent a triple blind, placebo-controlled crossover study evaluating two dose levels of methylphenidate (0.3 mg/kg and 0.5 mg/kg [corrected], twice daily) and placebo. Forty-nine trials were completed. Nineteen were children with ADHD-MD, 44 had ADHD only. Behavior and functioning were assessed at home and at school. Treatment effects were assessed using the Abbreviated Symptom Questionnaire for Parents and Teachers. Children with ADHD-MD were more likely to have severe ADHD-combined type and other neurodevelopmental and behavioral problems. Both groups of children had a linear dose response to medication (placebo, low, high) and there was no evidence of a group by dose interaction or an overall group effect at home or school. The lack of group effect suggests that these children responded to medication like the other subgroups.     

Valproate is associated with new-onset oligoamenorrhea with hyperandrogenism in women with bipolar disorder.

BACKGROUND: Preliminary evidence suggests that valproate is associated with isolated features of polycystic ovarian syndrome (PCOS), while contradictory data support an association between epilepsy and PCOS. The development of PCOS features after initiation of valproate was therefore examined in women with bipolar disorder using a standardized definition of PCOS. METHODS: Three hundred women 18 to 45 years old with bipolar disorder were evaluated for PCOS at 16 Systematic Treatment Enhancement for Bipolar Disorder sites. A comparison was made between the incidence of hyperandrogenism (hirsutism, acne, male-pattern alopecia, elevated androgens) with oligoamenorrhea that developed while taking valproate versus other anticonvulsants (lamotrigine, topiramate, gabapentin, carbamazepine, oxcarbazepine) and lithium. Medication and menstrual cycle histories were obtained, and hyperandrogenism was assessed. RESULTS: Among 230 women who could be evaluated, oligoamenorrhea with hyperandrogenism developed in 9 (10.5%) of 86 women on valproate and in 2 (1.4%) of 144 women on a nonvalproate anticonvulsant or lithium (relative risk 7.5, 95% confidence interval [CI] 1.7-34.1, p = .002). Oligoamenorrhea always began within 12 months of valproate use. CONCLUSIONS: Valproate is associated with new-onset oligoamenorrhea with hyperandrogenism. Monitoring for reproductive-endocrine abnormalities is important when starting and using valproate in reproductive-aged women. Prospective studies are needed to elucidate risk factors for development of PCOS on valproate.     

血管内栓塞治疗未破裂脑动静脉畸形并发癫痫患者的预后分析

目的分析血管内栓塞治疗未破裂脑动静脉畸形(CAVM)并发癫痫患者的预后情况。方法选择2013年3月至2017年6月收治的符合诊断标准的CAVM并发癫痫发作患者49例为研究对象,分析血管内栓塞治疗后患者的临床症状、生活质量(QOLIE-31)改善情况。结果患者经血管内栓塞治疗后,QOLIE-31各项指标(除了药物影响)评分均明显提高,高于治疗前(P0.05);Spetzler-Martin分级与Engel分级的I~II级例数多于治疗前(P0.05),同时Spetzler-Martin分级I~II级生活质量评分(76.04±18.33)分明显高于III~V级的(65.65±16.76)分(P0.05);Engel分级I~II级的生活质量评分(75.25±17.78)分明显高于III~V级的(66.23±13.22)分(P0.05);血管内栓塞比例80%的生活质量总评分(78.37±18.87)分明显高于栓塞比例80%的(64.16±16.92)分(P0.05);术后患者的头疼症状中重度例数明显低于治疗前(P0.01);患者的NIHSS评分和MRS评分均明显低于治疗前,头疼症状的生活质量评分高于治疗前(均P0.05)。结论血管内栓塞能明显改善未破裂脑动静脉畸形并发癫痫患者的头疼症状、癫痫发作情况、神经功能缺损,提高血管内栓塞比例能够提高患者生活质量。