锂剂抑制新生鼠缺氧缺血后的细胞凋亡及炎症反应研究

目的探讨锂对新生鼠缺氧缺血后的神经保护作用及相关机制。方法 40只9日龄Wistar雄性大鼠建立缺氧缺血动物模型,术后给予相同剂量的氯化锂或生理盐水干预,缺氧缺血后3d灌注取脑,MAP-2免疫组化染色评估脑损伤体积,Caspase-3检测细胞凋亡,Iba-1,Galectin-3检测神经炎症反应。结果缺氧缺血后3d,锂盐治疗组脑组织脑损伤体积较对照组明显减少(P<0.01);生理盐水组动物大脑皮质Caspases-3阳性细胞数目为(32.5±5.37)个,而锂盐治疗组则为(17.3±4.46)个(P<0.01),锂盐治疗后海马DG区Caspase-3阳性细胞数目仅为对照组的约1/4(P<0.001);Iba1染色,生理盐水组动物皮层及海马DG区Iba1阳性细胞数目分别为112.6±10.72和342.5±9.67,均显著高于锂盐治疗组(皮层72.4±7.33,海马DG区224.6±9.34)(P<0.01,P<0.01);Galectin-3染色,锂盐治疗后,实验动物大脑皮质及海马DG区Galectin-3阳性细胞数目均明显少于生理盐水组(P<0.05,P<0.01)。结论锂对新生鼠缺氧缺血具有显著的神经保护作用,其机制可能为减少细胞凋亡或抑制缺氧缺血后的神经炎症。     

丹参注射液对缺氧神经干细胞凋亡和Caspase-3活性的影响

目的 探讨丹参注射液对缺氧培养大鼠神经干细胞的凋亡及Caspase-3活性的影响,以进一步明确丹参注射液神经保护作用的分子机制.方法 体外培养新生大鼠海马神经干细胞,将其分为正常对照组,缺氧培养组及丹参注射液处理组.Hoechst染色后荧光显微镜下观察并计算细胞凋亡率:比色法检测各组细胞Caspase-3的相对活性.结果 缺氧培养大鼠神经干细胞的细胞凋亡率(30.12%±2.09%)及Caspase-3活性(3.85±0.41)均较正常对照组(2.75%±0.28%,1.16±0.07)明显升高,差异有统计学意义(P＜0.05);施加丹参注射液后,大鼠神经干细胞的细胞凋亡率(9.16%±1.34%)和Caspase-3活性(1.50±0.09)均较缺氧培养组明显下降,差异有统计学意义(P＜0.05).结论丹参注射液可对抗缺氧损伤所致的神经干细胞凋亡,从而起到神经保护作用.     

TNP-470联合化疗抑制胶质母细胞瘤的实验研究

目的 探讨TNP-470联合化疗药物卡氮芥(BCNU)对人U-251胶质母细胞瘤细胞裸鼠皮下移植瘤生长的作用.方法 将人U-251胶质母细胞瘤细胞株注射至裸鼠皮下,第7天荷瘤裸鼠随机分为4组:TNP-470治疗组、BCNU治疗组、TNP-470和BCNU联合治疗组、对照组.测体质量及肿瘤大小,以山羊抗小鼠CD105多克隆抗体免疫组化染色计数肿瘤微血管密度(MVD).结果 治疗后第21天联合治疗组移植瘤体积[(108.93±17.63)mm3]明显小于TNP-470治疗组[(576.10±114.29)mm3]及BCNU治疗组[(473.01±48.04)mm3](P均＜0.01);各治疗组移植瘤体积均小于对照组[(1512.61±470.25)mm3](P均＜0.01);TNP-470治疗组与BCNU治疗组间移植瘤体积差异无统计学意义(P＞0.05).治疗后第21天联合治疗组的抑瘤率(92.80%±11.37%)显著高于TNP-470治疗组(61.91%±6.29%)和BCNU治疗组(68.73%±9.65%)(P均＜0.01),TNP-470治疗组与BCNU治疗组间抑瘤率无统计学意义(P＞0.05).联合治疗组移植瘤MVD[(4.23 4±0.83)个/视野]明显低于TNP470治疗组[(5.70±0.85)个/视野]和BCNU治疗组[(8.60±0.87)个/视野](P均＜0.05);TNP-470治疗组移植瘤MVD显著低于BCNU治疗组(P＜0.05);各治疗组移植瘤MVD均较对照组[(11.32±1.50)个/视野]显著降低(P均＜0.05).结论 TNP-470联合化疗药物BCNU对人U-251胶质母细胞瘤细胞裸鼠皮下移植瘤的生长有显著抑制作用.     

利拉鲁肽对脑梗死小鼠保护作用的研究

目的探讨利拉鲁肽对实验性脑梗死小鼠的保护作用及分子机制。方法 C57BL/6小鼠随机分为假手术组、对照组和利拉鲁肽组。烧灼法建立永久、局灶性右侧皮质梗死模型(dMCAO),假手术组仅分离血管。利拉鲁肽组于dMCAO术后连续3d分别给予腹腔注射利拉鲁肽(200μg·kg-1·d-1),对照组腹腔注射等体积生理盐水。TTC染色方法计算脑梗死体积,Rota-rod方法评价肢体功能。术后72h时采用Western blot测定三个实验组p-AMPK/AMPK和NF-κB蛋白水平。结果利拉鲁肽明显延长dMCAO小鼠的Rota-rod时间(52.22±4.588 s),并缩小梗死体积(0.0791±0.0025),上调p-AMPK蛋白水平和p-AMPK/AMPK比值,抑制NF-κB表达。结论利拉鲁肽减轻dMCAO小鼠炎症反应和缺血性脑损伤,激活AMPK、抑制NF-κB是利拉鲁肽保护缺血性脑损伤的分子机制。     

百日咳毒素减轻脑缺血后神经元钙内流的实验研究

目的探讨百日咳毒素(PTx)对缺血性脑卒中后神经元的保护作用及其可能的机制。方法将C57BL6小鼠用大脑中动脉闭塞(MCAO)法建立卒中模型,随机分为两组,每组12只鼠。实验组卒中后予PTx 1000 ng溶于1 ml生理盐水腹腔注射干预;对照组予1 ml生理盐水腹腔注射。24 h后行TTC染色检测梗死面积,并行免疫组化检测细胞凋亡。在体外培养原代神经元,用谷氨酸兴奋刺激模拟卒中后神经元损伤模型。用MTT及乳酸脱氢酶(LDH)释放实验检测PTx对谷氨酸兴奋刺激后神经元的存活和损伤情况。然后,检测PTx对谷氨酸诱导神经元钙离子内流的影响。结果 PTx治疗可减小卒中后小鼠脑梗死面积,使之由(51±11)%降至(34±8)%(P0.05)。免疫组化发现PTx可使脑内Caspase-3阳性细胞数由(677.7±117.8)个/mm2减少至(297.5±83.6)个/mm2(P0.05),且较少细胞凋亡。体外结果提示,PTx可增加谷氨酸刺激后神经元的存活率,使MTT吸光值由(0.618±0.06)提升至(1.1±0.12)(P0.05);同时,PTx还可减少LDH的释放,使吸光值由(1.31±0.11)降低至(0.76±0.08)(P0.05)。PTx可减缓和减少钙离子进入神经元,经PTx治疗后钙内流可由基础值的5倍降低至基础值的2~3倍,且钙离子内流达到半峰值浓度的时间也由(18.5±2.5)s延长至(85.4±10.2)s。结论百日咳毒素可减少卒中后钙离子内流到神经元,继而减少神经元损伤,减小梗死面积。     

过氧化小体增殖剂激活型受体γ激活剂对缺血再灌注脑组织的保护作用及炎性机制分析

目的 探讨过氧化小体增殖剂激活型受体γ(PPARγ)激活剂对缺血再灌注脑组织的保护作用及其炎性机制.方法 健康雄性SD大鼠分为假手术组、生理盐水干预组、小剂量吡格列酮(PPARγ激活剂)干预组、大剂量吡格列酮干预组.吡格列酮干预组在中脑动脉闭塞(MCAO)前3 d分别给予吡咯列酮,每日一次灌胃给药.剂量分别是:小剂量组为10 mg/kg,大剂量组为15 mg/kg.生理盐水干预组仅给予等量生理盐水.假手术组亦给予等量生理盐水.以缺血后24h作为观察时间点,对各指标进行比较分析.氯化三苯基四氮唑(TTC)染色测定脑梗死体积,生化法测定髓过氧化物酶(MPO)活性.结果 小剂量吡格列酮干预组的脑梗死体积[(147±14)mm3]及大剂量吡格列酮干预组脑梗死体积[(121±16)mm3]均较生理盐水干预组[(183±17)mm3]小;小剂量吡格列酮干预组的MPO[(0.148±0.027)U/g]及大剂量吡格列酮干预组MPO[(0.096±0.021)U/g]均比生理盐水干预组[(0.203±0.022)U/g]降低,并且上述指标均呈现出随吡格列酮剂量的增加而下调幅度增强的趋势(P＜0.05).结论 PPARγ激活剂应用后,可以减少缺血再灌注脑组织梗死体积及中性粒细胞的浸润.本研究提示调控炎性损伤路径可能是利用PPARγ激活剂对PPARγ这一靶点进行干预从而发挥抗脑缺血损伤的机制之一.     

胶质细胞源性神经营养因子对大鼠局灶脑缺血损伤的保护作用

目的 从胶质细胞源性神经营养因子(GDNF)对大鼠局灶脑缺血梗死灶、半胱氨酸蛋白酶(Caspase-3)表达、细胞凋亡等方面的影响,研究其对大鼠局灶脑缺血的作用及其机制。方法 健康雄性Wistar大鼠120只,随机分为GDNF组和生理盐水组,每组又分为假手术组、缺血oh、3h、6h、24h组,采用大脑中动脉线栓模型,于栓塞同时大鼠脑室内分别给予GDNF和生理盐水5μL。检测脑梗死体积百分比、Caspase-3的表达、细胞凋亡等改变。结果 GDNF组脑梗死体积比明显小于生理盐水组;神经元损伤明显轻于生理盐水组,特别是海马区神经元在GDNF组无明显损伤;GDNF组(Caspase-3表达和TUNEL染色阳性细胞数明显少于生理盐水组。结论 GDNF对大鼠局灶脑缺血有保护作用,抑制Caspase-3的表达和细胞凋亡是其保护机制之一。     

急性脑梗死患者血清可溶性E-选择素、可溶性L-选择素的表达及其与脑梗死体积的关系

目的 探讨急性脑梗死(ACI)患者外周血可溶性E(sE)-选择素、可溶性L(sL)-选择素表达的变化及其与脑梗死体积的关系.方法 采用酶联免疫吸附法(ELISA)测定30例ACI患者(发病＜72 h)血清sE-选择素、sL-选择素的表达水平,并与20名健康者作对照.应用MRI弥散加权成像(DWI)技术测量ACI患者急性期脑梗死的体积,分析其与血清sE-选择素及sL-选择素水平的关系.结果 ACI患者血清sE-选择素的表达水平[(10.49±8.70)ng/ml]显著高于正常对照组[(2.93±2.07)ng/ml](P＜0.01),而sL-选择素的表达水平[(4.29±2.22)ng/ml]显著低于正常对照组[(6.41±3.28)ng/ml](P＜0.01);血清sE-选择素、sL-选择素水平与脑梗死体积不相关.结论 ACI患者血清sE-选择素表达上调,sL-选择素表达下凋,但都与脑梗死体积不相关,两者可能参与了缺血性脑损伤的病理过程.     

核因子-κB活性抑制剂对癫(癎)大鼠的脑保护作用

目的 研究核因子-κB(NF-κB)活性抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)对癫(癎)大鼠的脑保护作用.方法 将36只雄性SD大鼠随机分为癫(癎)组(14只)、PDTC干预组(PDTC组,14只)和假手术组(8只).采用海马注射海人酸(KA)方法制作癫(癎)大鼠模型,PDTC组大鼠造模前30 min给于腹腔注射PDTC150 mg,/kg;观察各组大鼠癫(癎)发作的潜伏期和初次至第6次≥Ⅳ级发作的时间(发作严重程度).应用HE染色和免疫组织化学染色,观察各组大鼠海马CA3区残存神经元数和NF-κB的表达.结果 PDTC组大鼠癫(癎)发作潜伏期[(89.6±39.3)min]长于癫(癎)组[(67.5±22.9)min],但差异无统计学意义;PDTC组初次至第6次≥Ⅳ级发作的时间[(29.2±20.4)min]较癫(癎)组[(12.1±4.0)min]显著延长(P＜0.05);与癫(癎)组相比,PDTC组大鼠海马CA3区残存神经元数显著增多(P＜0.05),NF-κB表达水平显著降低(P＜0.01),二者间呈负相关(r=-0.562,P=0.001).结论 NF-κB活性抑制剂能降低癫(癎)发作严重程度,减少海马神经元的变性死亡,具有脑保护作用.提示癫(癎)发作所致脑组织损伤可能与NF-κB活化有关.     

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。     

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。     

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.     

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.

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.     

Effects of Agonists and Antagonists of Cholecystokinin on Contractile and Myoelectric Activity of Isolated Muscle of Colon and Ileum in the Dog and Guinea Pig

This study evaluates the effects of agonists and antagonists of cholecystokinin (CCK) on contractile and myoelectrical activity in isolated longitudinal muscle strips from colon or ileum of guinea pigs or beagle dogs. Caerulein and CCK-8 caused a dose-dependent increase of contractile and myoelectrical spike activity in both species with maximal effects seen between 10⁻⁸ and 3 × 10⁻⁸ M. The dose responses were identical for both CCK agonists and species. The dose-related effects of CCK compounds on colonic muscle were slightly shifted to the right when compared to ileum in both species. All antagonists, the proglumide-derivatives CR1409, CR1392, and CR1505, as well as the nonpeptide substances asperlicin and L-364,718, caused a parallel rightward shift of CCK's dose-dependent motor activity response, indicating the competitive nature of inhibition. The antagonists displayed a rank order of potency in antagonizing CCK's action on intestinal motility similar to their ability to antagonize CCK's action on pancreas and gallbladder. L-364,718 was the most potent antagonist, followed by CR1409, CR1505, CR1392, asperlicin, and proglumide. The antagonists did not affect contractile or myoelectrical responses to acetylcholine, histamine, motilin, or substance P. Thus compounds that have been described as CCK antagonists for pancreas and gallbladder also act as specific and competitive antagonists of CCK's action on contractile and myoelectrical activity of Heal and colonie muscle.     

The neuroendocrinology of stress and the pathophysiology and therapy of depression and anxiety

Clinical and preclinical studies have gathered substantial evidence that stress response alterations play a major role in the development of major depression, panic disorder, and post-traumatic stress disorder. The stress response, the hypothalamic pituitary adrenocortical (HPA) system and its modulation by corticotropin-releasing hormones (CRH),corticosteroids,and their receptors, and the roles of natriuretic peptides and neuroactive steroids are described. We review the role of the HPA system in major depression, panic disorder, and post-traumatic stress disorder and its possible relevance for treatment. Impaired glucocorticoid receptor function in major depression is associated with an excessive release of neurohormones such as CRH, to which a number of signs and symptoms characteristic of depression can be ascribed. In panic disorder, a role of central CRH in panic attacks has been suggested. Atrial natriuretic peptide (ANP) is causally involved in sodium lactate-induced panic attacks. Furthermore, preclinical and clinical data on its anxiolytic activity suggest that nonpeptidergic ANP receptor ligands may be potentially useful in the treatment of anxiety disorders. Post-traumatic stress disorder is characterized by a peripheral hyporesponsive HPA system and elevated CRH concentrations in the CSF. This dissociation is probably related to an increased risk of this disorder. We further review recent data that describe an important role of GABA(A)-receptor modulatory,3 alpha-reduced neuroactive steroids in major depression, anxiety, and its treatment. Antidepressants are effective in both depression and anxiety disorders and have major effects on the HPA system,especially on glucocorticoid and mineralocorticoid receptors. Normalization of HPA system abnormalities is a strong predictor of the clinical course, at least in major depression and panic disorder. Currently,CRH-R1 or glucocorticoid receptor antagonists and ANP receptor agonists are being studied and may provide future treatment options more closely related to the pathophysiology of these disorders.     

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.     

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.