脑创伤后bcl—2蛋白的神经保护作用

目的 探讨液压脑损伤后凋亡抑制基因bcl—2的变化规律及bcl—2基因在创伤性脑损伤后细胞凋亡中的作用。方法 应用免疫组化观察大鼠中型液压脑损伤伤前及伤后6h、12h、1d、3d、7dbcl—2蛋白表达情况,应用TUNEIL和电镜观察伤后细胞死亡的形态。结果 免疫反应阳性细胞主要位于伤侧大脑半球皮质、皮层下白质、海马CAl、CA3及齿状回的神经元和神经胶质细胞,以海马CA3区最为显。在高倍镜下,表达Bcl—2蛋白的神经细胞胞核形态正常,很少见到凋亡或坏死的形态特征。伤后早期(6h),打击侧海马CA3区Bcl—2蛋白表达显下降;Bcl—2早期改变出现在伤后6h,比细胞凋亡提前表现;伤后l—3h,Bcl—2的表达下降相对缓慢。结论 bcl—2蛋白在抑制脑创伤后细胞凋亡中起重要作用,bcl—2可能是一种可诱导的神经保护因子。     

大鼠SAH后的蛛网膜颗粒细胞凋亡及血清TNF-α表达

目的通过动物实验分析大鼠蛛网膜下腔出血(SAH)后的蛛网膜颗粒细胞凋亡及血清TNFα表达,为研究SAH后脑积水的病因及预防提供理论依据。方法采用小脑延髓池注血法建立实验组大鼠SAH动物模型,并分为B、C、D、E组,每组8只。对照组小脑延髓池内注入生理盐水。分别于1d(B组)、4d(C组)、7d(D组)、10d(E组)处死实验组动物,于10d处死对照组(A组)动物。取血清测定肿瘤坏死因子α(TNFα)含量;采用TUNEL法检测蛛网膜颗粒的细胞凋亡。结果A组蛛网膜颗粒中偶见细胞凋亡,B、C、D、E4组蛛网膜颗粒中凋亡细胞逐渐增多。蛛网膜细胞、内皮细胞及纤维细胞3类细胞均有凋亡现象。A组大鼠血清中含有低水平TNFα,蛛网膜下腔注血后大鼠血清中的TNFα升高,呈双峰样改变,C、E组即第4、10d为顶点。细胞凋亡指数(AI)与血清TNFα含量呈正相关(r=0.315,P<0.05)。结论SAH后过度的炎症反应可能导致蛛网膜颗粒出现过度的细胞凋亡以及蛛网膜颗粒功能减退;蛛网膜颗粒的细胞凋亡可能与高水平TNFα有关。     

Gap19介导Cx43半通道抑制JAK2/STAT3通路活化对急性脑梗死大鼠的保护作用

目的探究Gap19通过介导Cx43半通道活性和JAK2/STAT3通路对急性脑梗死(ACI)大鼠的影响。方法 24只SD大鼠随机分为3组:对照组、MCAO组和MCAO+Gap19组,每组8只。建立ACI大鼠模型(MCAO)和氧葡萄糖剥夺(OGD)体外模型。TTC染色检测大鼠脑梗死面积;mNSS法检测大鼠神经功能缺损;CCK-8检测星形胶质细胞活力;Western blot检测NVU模型Cx43、Cleaved caspase-3、Bcl、Bax和JAK2/STAT3通路相关基因的蛋白表达;溴化乙锭摄取实验检测星形胶质细胞中半通道活性;免疫荧光法检测星形胶质细胞中pSTAT3和Cx43的共定位。结果 MCAO处理导致大鼠脑梗死面积和神经功能缺损均显著上调(P 0.05),Gap19处理显著下调大鼠脑梗死面积和神经功能缺损(P 0.05)。OGD处理导致NVU模型Cx43的蛋白表达和半通道活性显著上调(P 0.05),细胞活力显著下调(P 0.05),促进细胞凋亡;Gap19处理显著上调OGD处理后NVU模型细胞活力(P 0.05),显著下调Cx43的蛋白表达和星形胶质细胞的半通道活性(P 0.05),抑制细胞凋亡,促进JAK2/STAT3通路活化,其中Cx43与pSTAT3存在共定位关系。结论 Gap19通过抑制Cx43半通道和活化JAK2/STAT3通路发挥对ACI大鼠的保护作用。     

大鼠脑缺血-再灌注后小脑AQP4蛋白的表达变化

目的观察大鼠大脑中动脉闭塞后远隔部位小脑组织形态学改变以及水通道蛋白—4(AQP4)的表达变化。方法健康雄性SD大鼠40只,体质量260~300g,随机分成4组,分别为脑缺血—再灌注12h、1d、3d组和假手术组,采用线栓法制作大鼠右侧大脑中动脉脑缺血再灌注模型,Longa评分法对大鼠进行神经功能评分,苏木精—伊红染色观察大鼠小脑的组织形态学变化;免疫组化染色测定大鼠小脑AQP4蛋白表达,测量平均吸光度值(mean optical density,MOD)评价染色程度。结果假手术组动物麻醉苏醒后未见神经功能缺损。脑缺血—再灌注12h神经功能评分升高,12h~1d之间无明显差异,3d神经功能评分减低(P0.05)。苏木精—伊红染色显示假手术组小脑结构未见明显异常,脑缺血—再灌注后12h小脑区出现缺血性损害,细胞肿胀,胞核深染,但细胞排列尚好;1~3d时胞核浓染加深,出现核固缩,细胞排列紊乱,间隙增宽。假手术组小脑区AQP4表达极少,脑缺血—再灌注12h、1d和3d大鼠缺血侧小脑区均可见AQP4阳性着色细胞,12h~1d AQP4 MOD值随时间延长逐渐增高,3d时较1d时降低,3组AQP4表达水平均明显高于假手术组(P0.05)。结论大脑中动脉脑梗死后小脑区存在损伤,并且有AQP4表达增加,小脑区AQP4表达同神经功能损伤程度密切相关。     

艾司西酞普兰对抑郁大鼠海马BDNF表达及细胞凋亡的影响

目的观察艾司西酞普兰对抑郁大鼠海马BDNF表达及细胞凋亡的影响,探讨其作用机制。方法40只雄性SD大鼠随机分为对照（A）、对照＋药（B）、抑郁模型（C）和抑郁模型＋药（D）组共4组,采用慢性不可预见性的温和刺激结合孤养法建立抑郁大鼠模型,TUNEL法检测海马细胞凋亡情况,实时荧光定量PCR方法检测海马BDNF、Bax、Bcl2、Caspase3mRNA的表达。结果（1）C组海马细胞凋亡数显著增加,与A组比较差异有统计学意义（P〈0．01）,而B组细胞凋亡数与A组差异无统计学意义（P〉0．05）;D组细胞凋亡数明显减少,与c组比较差异有统计学意义（P〈0．01）。（2）C组海马BDNF、Bel—2mRNA表达降低,Bax、Caspase3mRNA表达升高,与A组比较差异均有统计学意义（P〈0．01）;D组BDNF、Bcl2mRNA表达增加,Bax、Caspase3mRNA表达降低,与C组比较差异均有统计学意义（P〈0．01）。结论艾司西酞普兰可改善抑郁大鼠的抑郁行为及海马细胞凋亡,其机制可能是通过增加海马BDNFmRNA的表达,上调Bcl一2及下调Bax、Caspase3mRNA的表达,从而预防海马细胞凋亡而发挥脑保护的作用。     

17-β雌二醇对大鼠脑缺血再灌注损伤细胞凋亡影响的实验研究

目的观察雌二醇对大鼠脑缺血再灌注损伤脑细胞凋亡的影响。方法72只大鼠随机分为假手术组（n=8）、实验对照组及雌二醇治疗组，后两组又进一步分为3h、6h、12h、24h4个时间点，每时间点8只。线栓法建立大鼠局灶性脑缺血再灌注损伤模型，石蜡切片HE染色及免疫组化染色检测脑组织的细胞凋亡情况及凋亡调控基因Bcl-2、Caspase-3的表达。结果雌二醇治疗组的脑组织缺血半暗带区细胞凋亡较实验对照组明显减少；随着再灌注时间的延长，治疗组半暗带区Bcl一2的表达上调而Caspase-3的表达上调减弱。结论17-β雌二醇具有减少脑缺血再灌注损伤细胞凋亡的作用，而凋亡抑制基因Bcl-2的表达升高及凋亡执行蛋白Caspase-3的表达减弱可能其是重要机制之一。     

蛛网膜下腔移植骨髓基质干细胞治疗大鼠脊髓损伤模型Fas、Fas—L表达变化

目的 通过蛛网膜下腔移植绿色荧光蛋白转基因小鼠骨髓基质干细胞治疗大鼠脊髓挫伤模型,观察凋亡因子Fas和Fas-L在移植后不同时点的变化情况，为骨髓基质干细胞治疗脊髓损伤及机制提供实验室依据。方法 （1）成年健康SD雌鼠66只（体重200～250g）随机分成11组，每组6只，即空白对照组、损伤对照组、细胞移植治疗组（后2组再分为术后1、3、7、14、21d组）。（2）在手术前、后对大鼠进行BBB评分和爬网格试验。术后第2d移植MSCs。之后，制作15～20μm厚的连续冰冻切片，间隔取片进行荧光细胞观察及Fas和Fas—L抗体免疫组织化学ABC染色。并在光学显微镜下对脊髓灰质前角进行阳性细胞计数，用图文分析系统检测Fas和Fas—L免疫阳性反应物的平均灰度值。应用SPSS11．0软件包进行组间多个样本均数比较的单因素方差分析、q检验。结果 3组脊髓组织中均可见Fas和Fas—L阳性细胞。损伤对照组和细胞移植治疗组Fas和Fas—L阳性细胞数出现先升高后降低的趋势，1d组就出现表达，持续到14d组，21d组表达接近正常对照组，高峰出现在7d。21d接近正常对照组。损伤对照组与细胞移植治疗组相比较，阳性细胞数在7d、14d组出现差异（P〈0．05）细胞移植治疗组Fas和Fas—L1、3、7d组灰度值低于正常对照组（P〈0．05）。结论 与损伤对照组相比细胞移植治疗组7、14d组Fas、Fas-L阳性细胞数下降，灰度值升高，2组间有显著性差异，说明移植骨髓基质干细胞下凋7d及14d组Fas、Fas-L的表达。     

实验性癫痫不同脑区细胞凋亡的观察及意义

目的 观察大鼠杏仁核点燃癫痫后细胞凋亡情况。探讨其在点燃癫痫后不同脑区不同点燃时程时变化的意义。方法 建立大鼠杏仁核点燃癫痫模型。应用流式细胞术（FCM）检测额叶，颞叶及海马神经细胞中DNA状态，同时应用TUNEL染色方法观察以上部位凋亡细胞变化。结果 大鼠燃后1d即有凋亡细胞出现，点燃3d后凋亡细胞明显增多，并在1周时达高峰，点燃后21d及49d后，凋亡细胞有所减少，但仍高于正常水平，与对照组相比有显著意义。结论 凋亡现象贯穿于癫痫鼠病程的始终，与癫痫发作密切相关。     

小鼠小脑皮质的生长发育

目的：探讨小鼠小脑皮质的组织发生过程。方法：应用光镜和电镜技术对胚胎和生后小脑皮质进行形态学观察,对各层厚度和细胞密度进行测量。结果：胚胎12d（E12）小脑原基有室管膜层、套层和边缘层构成,约出生当日（P0）出现外颗粒层、分子层、Purkinje细胞层和内颗粒层。外颗粒层P6／7达最厚,至P20消失。P0至P30内颗粒层细胞逐步分化发育成熟,Purkinje细胞树突树逐渐形成,约P7时Purkinje细胞排列成单层。结论：E12至P0片层化小脑主要经历了细胞增殖、分化与迁移;P0至P30片层化结构逐渐发育成熟,外颗粒层消亡以细胞迁移和凋亡为主,其他各层细胞主要经历了分化发育与凋亡。     

CNTF多克隆抗体的制备与初步应用

以重组表达并经纯化的人睫状神经营养因子（hCNTF）免疫家兔，得到抗ACNTF抗血清。经硫酸铵沉铁、proteinA亲和层析后得到纯化的抗hCNTF多克隆抗体。应用该抗体对大鼠脑组织进行免疫组化研究，结果发现：CNTF抗体染色阳性物不仅存在于各脑区胶质细胞中，而且也存在于皮质的各层神经元，小脑分子层神经元、蒲肯野细胞、小脑颗粒细胞，海马锥体细胞、齿状回勤粒细胞及面神经核细胞中。CNTF祥物质不仅存在于神经元胞浆中，也存在于胞核中。     

Cultural Identity and Experiences of Prejudice and Discrimination of Afghan and Iranian Immigrant Youth

In culturally diverse and immigrant receiving societies, immigrant youth can be subject to prejudice and discrimination. Such experiences can impact on immigrant youth’s cultural identity and influence their psychosocial outcomes. This paper presents findings of a study that examined cultural identity and experiences of prejudice and discrimination among Afghan (N = 9) and Iranian (N = 17) immigrant youth in Canada. The study had a prospective, comparative, longitudinal qualitative design. Data was gathered through focus groups, interviews, journals and field logs. Four main themes emerged on participants’ experiences of prejudice and discrimination: (a) societal factors influencing prejudice; (b) personal experiences of discrimination; (c) fear of disclosure and silenced cultural identity; and (d) resiliency and strength of cultural identity. Drawing from Rosenberg’s (Conceiving the self, Basic Books, New York, 1979) self-concept framework and Romero and Roberts (J. Adolesc., 21:641–656, 1998) distinction between prejudice and discrimination, results indicated that youth’s extant and presenting cultural identity were affected. Inclusive policies and practices are needed to promote youth integration in multicultural and immigrant receiving settings.

Efficacy and Effectiveness of Child and Adolescent Psychotherapy and Pharmacotherapy

Decades of intervention research have produced a rich body of evidence on the effects of psychotherapies and pharmacotherapies with children and adolescents. Here we summarize and critique that evidence. We review findings bearing on the efficacy of psychosocial treatments and medications under controlled experimental conditions. We also report evidence, where available, on the effectiveness of both classes of treatment with clinically referred youth treated in real-world clinical contexts. In general, the large body of evidence on efficacy contrasts sharply with the small base of evidence on effectiveness. Addressing this gap through an enriched research agenda could contribute importantly to linking scientific inquiry and clinical practice—to the benefit of both ventures. This is one element of a multifaceted agenda for future research and for synthesis of research, which will require the interplay of multiple disciplines related to child and adolescent mental health.     

癫痫共病抑郁的机制及临床诊疗

本文目的是探讨癫痫共病抑郁的可能机制及临床诊疗。癫痫是一种常见的、慢性的、致残性的神经疾病,癫痫患者生活质量下降,存在明显的负性情绪,常伴发各种精神疾病。癫痫与抑郁具有共同的神经生物学基础,可能存在共同的发病机制。本文从癫痫共病抑郁的发病机制、临床诊断及治疗方面予以总结归纳。     

沙盘游戏疗法在地中海贫血患儿心理干预中的应用

本文目的是对沙盘游戏疗法在地中海贫血患儿心理干预中的应用进行综述,以期为地中海贫血患儿的心理康复提供参考。地中海贫血是以珠蛋白生成障碍为主要特征的遗传性疾病,由于长期输血治疗,患儿存在较多的心理和行为问题。沙盘游戏疗法作为一种有效、实用的儿童心理治疗方法,对提高地中海贫血患儿的康复效果、改善生存质量有重要的临床意义。     

小胶质细胞和少突胶质细胞前体的培养和鉴定

目的 探讨新生大鼠脑组织小胶质细胞(MG)和少突胶质细胞(OL)前体的分离和体外培养方法 . 方法 取新生2 d SD大鼠脑组织,体外原代培养混合胶质细胞7 d后,分别采用"改良振荡伴差速贴壁"法和"营养缺失伴振荡"法纯化培养MG和OL前体,并分别应用免疫荧光染色异凝集素-B4(IB4)和OL前体标记物(O4)进行鉴定.结果 混合胶质细胞培养7 d后呈明显三层增长,其中MG位于上层,星型胶质细胞位于底层,两者之间为2型少突星型(O2A)祖细胞.纯化培养后OL前体胞体呈小圆形,有双极或三极突起,MG则以阿米巴形、圆形居多,或边缘呈毛刺状.免疫荧光染色IB4显示绿色荧光,MG纯度达到90%以上.免疫荧光染色O4显示棕黄色荧光,OL前体纯度达到95%以上. 结论 采用"改良振荡伴差速贴壁"法以及"营养缺失伴振荡"法分别成功获取大量纯度高、活力好的MG和OL前体.     

发作性睡病与异态睡眠的诊治

本文目的是探讨发作性睡病与异态睡眠的诊断与治疗。发作性睡病被漏诊和误诊的几率较高,危害较大,共患异态睡眠比例高。文章从发作性睡病临床特征、REM睡眠的作用、发作性睡病与异态睡眠(睡眠瘫痪、睡眠幻觉、快眼动睡眠期行为障碍)共病特征及治疗这四个方面进行了讨论。     

Neurological and neuropsychological consequences of electrical and lightning shock:review and theories of causation

Injuries from lightning and electrical injuries involve multiple systems of the body, however neurological symptoms are very widely reported. A disabling neuropsychological syndrome is also noted.This paper presents a comprehensive review of neurological and neuropsychological symptoms. Partial theories of causation for these injuries have been advanced, however, there is no convincing explanation for both delay in onset of symptoms and also the genesis of the neuropsychological syndrome. A theory of causation is proposed which satisfies both these constraints.This theory suggests circulating hormones such as cortisol, together with nitric oxide and oxidant free radicals from glutamatergic hyper-stimulation, act on tissues remote from the injury path including the hippocampus.This theory opens a research path to explore treat-ment options.     

Review and meta-analysis of the phenomenology and clinical characteristics of mania in children and adolescents

OBJECTIVE: Using predetermined criteria for study quality and methods, a literature review and meta-analysis of seven reports about pediatric bipolar disorder (BPD) was conducted to determine if there is a consistent picture of the phenomenology and clinical characteristics of BPD in children and adolescents. METHODS: Searches were conducted in MedLine and PsycINFO using the terms mania, BPD, children and adolescents, and was limited to published articles in peer-reviewed journals. Seven reports were selected that met the following criteria: a systematic method for the elicitation and reporting of symptoms and clinical characteristics of subjects; subjects were interviewed by a trained researcher or clinician; ages 5-18 years; use of a diagnostic system, either DSM or RDC for categorization; a consensus method for the establishment of the diagnosis of BPD. RESULTS: Most DSM-IV symptoms of mania were common in the children and adolescents with BPD with the most common symptoms being increased energy, distractibility, and pressured speech. On average, four of five bipolar cases also showed threshold levels of irritable mood and grandiosity, and more than 70% of all cases showed elated/euphoric mood, decreased need for sleep, or racing thoughts. Roughly 69% of cases also showed poor judgment, whereas only half of bipolar cases demonstrated flight of ideas, and slightly more than one-third showed hypersexuality or psychotic features. CONCLUSIONS: The clinical picture that emerges is that of children or adolescents with periods of increased energy (mania or hypomania), accompanied by distractibility, pressured speech, irritability, grandiosity, racing thoughts, decreased need for sleep and euphoria/elation.     

Design and pharmacological activity of glycinamide and N-methoxy amide derivatives of analogs and constitutional isomers of valproic acid

A series of glycinamide conjugates and N-methoxy amide derivatives of valproic acid (VPA) analogs and constitutional isomers were synthesized and evaluated for anticonvulsant activity. Of all compounds synthesized and tested, only N-methoxy-valnoctamide (N-methoxy-VCD) possessed better activity than VPA in the following anticonvulsant tests: maximal electroshock, subcutaneous metrazol, and 6-Hz (32-mA) seizure tests. In mice, the ED₅₀ values of N-methoxy-VCD were 142 mg/kg (maximal electroshock test), 70 mg/kg (subcutaneous metrazol test), and 35 mg/kg (6-Hz test), and its neurotoxicity TD₅₀ was 118 mg/kg. In rats, the ED₅₀ of N-methoxy-VCD in the subcutaneous metrazol test was 36 mg/kg and its protective index (PI = TD₅₀/ED₅₀) was > 5.5. In the rat pilocarpine-induced status epilepticus model, N-methoxy-VCD demonstrated full protection at 200 mg/kg, without any neurotoxicity. N-Methoxy-VCD was tested for its ability to induce teratogenicity in a mouse strain susceptible to VPA-induced teratogenicity and was found to be nonteratogenic, although it caused some resorptions. Nevertheless, a safety margin was still maintained between the ED₅₀ values of N-methoxy-VCD in the mouse subcutaneous metrazol test and the doses that caused the resorptions. On the basis of these results, N-methoxy-VCD is a good candidate for further evaluation as a new anticonvulsant and central nervous system drug.     

Polymorphisms of DRD4 and DRD3 and risk of avoidant and obsessive personality traits and disorders

We investigated whether polymorphisms of the dopamine D4 receptor (DRD4) and polymorphisms of the dopamine D3 receptor (DRD3) were associated with personality disorder symptomatology rather than with personality traits such as novelty seeking. DNA was obtained from 145 depressed patients in a clinical trial. These patients were assessed for the presence of personality disorder symptoms and disorders. The 2-repeat allele of the DRD4 exon III polymorphism was associated with increased rates of avoidant and obsessive personality disorder symptomatology. The T,T genotype of the DRD4 -521 C>T polymorphism was also associated with increased rates of avoidant and obsessive personality disorder symptomatology. The Gly9,Gly9 genotype of the DRD3 Ser9Gly polymorphism was associated with increased rates of obsessive personality disorder symptomatology. None of these three polymorphisms were associated with novelty seeking or other temperament traits on the Temperament and Character Inventory. Our results suggest that genetic polymorphisms of DRD4 and DRD3 may well be associated with personality traits, and that conflicting findings to date may arise from the problem of phenotype definition.