脑卒中后抑郁大鼠海马BDNF及其受体TrkB蛋白的表达变化

目的:探讨脑卒中后抑郁(PSD)大鼠海马脑源性神经营养因子(BDNF)及其高亲和力受体酪氨酸激酶B(TrkB)蛋白的表达变化。方法:24只雌性SD大鼠随机分为正常组、抑郁组、脑卒中组及PSD组,每组6只。采用线栓法建立局灶性脑缺血(卒中)模型;采用孤养方法建立抑郁模型;局灶性脑缺血模型加以慢性不可预见的中等应激刺激和孤养法建立PSD模型;并设正常组做为正常对照。应用免疫组化检测各组大鼠造模后第29天大鼠海马BDNF和TrkB免疫阳性细胞数的表达变化。结果:PSD组海马BDNF免疫阳性细胞数最少[(8.56±1.67)个/视野]。PSD组、抑郁组及脑卒中组与正常对照组相比BDNF阳性细胞数均明显减少(P0.05);PSD组海马TrkB免疫阳性细胞数最少[(6.44±1.13)个/视野],与各对照组相比,差异均有统计学意义(P0.05)。结论:PSD大鼠海马BDNF及其高亲和力受体TrkB蛋白表达减少可能在PSD发病过程中发挥了一定的作用。     

shRNA干扰蓝斑核去甲肾上腺素转运体(NET)表达对大鼠抑郁样行为的改善作用

目的:探讨利用慢病毒在蓝斑核(locus coeruleus,LC)敲除去甲肾上腺素转运体(norepinephrine transporter,NET)对大鼠抑郁样行为的影响。方法:构建NET-shRNA慢病毒载体及阴性对照序列,感染PC12细胞后通过Q-PCR和Western Blot方法测定NET mRNA和蛋白水平;建立慢性不可预见性温和应激大鼠抑郁症模型,双侧蓝斑核注射NET-shRNA;3周后采用强迫游泳实验、糖水偏好实验、旷场实验检测大鼠抑郁样行为;Western Blot检测大鼠NET水平;高效液相色谱法(HPLC)检测大鼠去甲肾上腺素(norepinephrine,NE)水平。结果:Q-PCR和Western Blot检测结果显示:shRNA-1146沉默效率最高(P0.01);与抑郁症模型组相比,蓝斑核注射NET-shRNA组大鼠强迫游泳不动时间减少、糖水偏好率增加、旷场实验得分增加(P0.01),蓝斑核NET表达下降(P0.01)、NE水平升高(P0.05)。结论:NET-shRNA慢病毒注射蓝斑核能下调NET的表达,改善大鼠抑郁样行为,为抑郁症的基因治疗提供了实验依据。     

大鼠海马脑区脑源性神经营养因子慢病毒注射与表达的检测

目的:建立成年大鼠海马脑区注射脑源性神经营养因子(BDNF)慢病毒表达载体的方法,并检测BDNF慢病毒载体体内表达目的基因的能力.方法:健康雄性SD大鼠随机分为假手术组(SH)、生理盐水组(NS)、GFP慢病毒注射组(GFP)和BDNF慢病毒注射组(BDNF),每组15只.所有实验大鼠在脑立体定位仪下定位海马组织.NS、GFP和BDNF组分别给予盐水(2μL)、GFP慢病毒(2μL)和BDNF慢病毒(2μL),SH组只实行手术操作.注射手术7d、14d和30d后,分别处死大鼠并进行在冰上取各组大鼠海马组织,分别提取海马总RNA和总蛋白质通过RT-PCR和Westernblot检测各组大鼠海马组织BDNFmRNA和蛋白表达水平的变化.同时通过原位杂交方法检测BDNF的表达水平和脑区分布.结果: BDNF慢病毒注射成年大鼠7d、14d和30d后,均可以成功检测到BDNF在mRNA和蛋白水平在海马脑区都有明显的表达,同假手术组相比具有显著性差异;原位杂交检测显示BDNF慢病毒注射成年大鼠海马脑区7d、14d和30d的BDNF表达水平和脑区分布均保持稳定.结论:成功地建立了BDNF慢病毒注射成年大鼠海马脑区的方法,并可在注射后不同时间段检测到BDNF高水平的表达和均匀的海马脑区分布,为进一步将其应用于神经系统损伤性疾病的治疗奠定了基础.     

解郁丸对WKY大鼠的抑郁样行为及海马和前额叶皮层BDNF表达的影响

目的:观察解郁丸对Wistar-Kyoto(WKY)大鼠抑郁样行为及脑源性神经营养因子(BDNF)在海马和前额叶皮层表达的影响,探讨其抗抑郁作用及相关机制。方法:成年雄性WKY大鼠为内源性抑郁动物模型,选取同品系Wistar大鼠作为空白对照组,WKY大鼠随机分为模型组、西酞普兰组和解郁丸组,分别灌胃给药21 d后,用糖水偏好实验及强迫游泳实验观察各组大鼠抑郁行为变化;采用免疫荧光法和Western blot法检测海马及前额叶皮层BDNF表达水平的变化。结果:WKY大鼠表现出明显的抑郁样行为,海马及前额叶皮层的BDNF表达量显著下降,且海马区神经元轴突减少(P0.01);在药物治疗后,WKY大鼠的抑郁样行为明显减少,BDNF在海马及前额叶皮层中的表达增加,且轴突数目也增加(P0.01)。结论:解郁丸能有效减少WKY大鼠的抑郁样行为;BDNF是其抗抑郁作用发挥的关键因子。本研究也进一步验证BDNF参与抑郁的发生发展过程。     

脑室注射BDNF对抑郁症大鼠海马神经元前体细胞微管相关蛋白Doublecortin表达的影响

目的 探讨脑源性神经营养因子(brain-drived neurotrophic factor,BDNF)对抑郁症大鼠海马神经元前体细胞微管相关蛋白Doublecortin(DCX)表达的影响,为BDNF如何影响神经元前体细胞的增殖、迁移提供实验依据.方法 成功建立抑郁症大鼠模型后,向脑室注射0.5 μg BDNF,取脑组织行冰冻切片.采用免疫组织化学和免疫荧光技术观察DCX阳性细胞表达情况及表达部位.结果 相对于未注射BDNF的抑郁症大鼠,注射组大鼠海马区域DCX阳性细胞数明显增多,且与未注射组存在明显差异(P＜0.05).结论 BDNF可能通过调控抑郁症大鼠海马神经元前体细胞微管相关蛋白DCX的表达,从而影响抑郁症时神经元前体细胞的分化和迁移.     

环磷酸腺苷反应元件结合蛋白调控耐药性癫痫大鼠海马内脑源性神经营养因子的表达

背景：耐药性癫痫的形成机制尚不清楚,其中环磷酸腺苷反应元件结合蛋白(cAMP response element-binding protein,CREB)及脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)在耐药性癫痫形成过程中的作用已被广泛研究。目的：使用慢病毒转染法调控正常大鼠海马CREB表达水平,探究其对耐药性癫痫形成过程及海马内BDNF表达的影响。方法：使用过表达慢病毒或干扰慢病毒调控大鼠海马中CREB表达水平,检测病毒转染结果;大鼠注射病毒7 d后构建氯化锂-匹罗卡品癫痫模型,随后使用苯妥英钠和苯巴比妥进行耐药性筛选,筛选出耐药性癫痫大鼠,分为过表达组、过表达对照组、干扰组、干扰对照组,观察筛药过程中大鼠癫痫发作次数变化,使用蛋白质免疫印迹法及实时荧光定量PCR法检测大鼠海马中CREB及BDNF表达水平。结果与结论：(1)病毒转染后14 d,大鼠海马区满布绿色荧光,病毒转染水平较好;(2)药筛前过表达组大鼠癫痫发作次数多于过表达对照组,干扰组大鼠癫痫发作次数少于干扰对照组;药筛后过表达组大鼠癫痫发作次数多于过表达对照组,干扰组大...     

槲皮素治疗联合运动训练对脑卒中后大鼠抑郁样行为影响的研究

目的:探讨槲皮素治疗联合运动训练对脑卒中模型大鼠抑郁样行为的治疗作用。方法:将50只大鼠随机分为假手术组(sham)、卒中抑郁组(PSD)、槲皮素组(QUE)、运动组(EXE)和槲皮素+运动组(QUE+EXE)。通过大脑中动脉闭塞(MCAO)技术制备大鼠卒中后抑郁模型。通过悬尾实验(TST)和糖水偏爱实验(SPT)来评估各组大鼠抑郁样行为;利用酶联免疫吸附试验(ELISA)检测各组大鼠血清和前额叶皮质中肿瘤坏死因子α(TNF-a)和白细胞介素1-β(IL-1β)的含量;利用Western Blot检测各组大鼠前额叶皮质中脑源性神经生长因子(BDNF)、原肌球蛋白受体激酶B (Trk-B)、糖原合成酶激酶-3β(GSK-3β)和β-连环蛋白(β-catenin)的表达。结果:行为学检测结果显示PSD组大鼠不动的时间显著增加(P<0.05),同时糖水摄入量及糖水偏爱百分比均显著下降(P<0.05),而槲皮素和运动训练均逆转了这些行为(P<0.05)。ELISA检测显示PSD组大鼠血清及前额叶皮质中TNF-α和IL-1β表达升高(P<0.05),而槲皮素和运动训练均不...     

抑郁大鼠海马微管相关蛋白pMAP-2和p-Tau表达的研究

目的:观察微管蛋白p MAP-2和p-Tau在抑郁大鼠海马的表达变化情况,探讨海马微管蛋白与抑郁症的关系和意义。方法:通过利用慢性不可预见性温和应激(chronic unpredicted mild stress,CUMS)建立大鼠抑郁症模型,对照组和模型组分别采用糖水偏好实验、旷场实验以及Morris水迷宫对其行为进行学检测,采用尼氏染色观察海马神经元的形态学变化,对照组和模型组造模后1、7、14、28 d分批处死大鼠,用Western Blot方法检测海马组织中p-MAP-2和p-Tau的蛋白表达水平,免疫荧光双标观察海马组织中p-MAP-2和p-Tau定位分布。结果:糖水偏好实验:模型组糖水消耗量和糖水偏好百分比分别低于对照组(P0.01);旷场实验:模型组行走总路程、中央活动时间、直立次数和修饰行为分别低于对照组(P0.01);Morris水迷宫模型组平均逃避潜伏期高于对照组(P0.01)。CUMS后1 d大鼠海马p-MAP-2和p-Tau蛋白的表达低于对照组,7 d后开始升高,14 d明显高于对照组,28 d仍略高于对照组。p-MAP-2和p-Tau免疫荧光双标结果显示:部分神经元的胞浆内p-MAP-2和pTau同时表达,表明p-MAP-2和p-Tau有部分共定位。结论:p-MAP-2和p-Tau在抑郁模型大鼠海马出现先降低后增高的现象,因此我们推测在应激抑郁症发病过程中,p-MAP-2和p-Tau蛋白表达的变化可能影响微管的稳定性,破坏细胞骨架稳态,从而影响海马神经元的结构改变。     

芬戈莫德对卒中后抑郁大鼠的治疗作用

目的:观察芬戈莫德(FTY720)对卒中后抑郁(PSD)大鼠的治疗效果并初步探索其作用机制。方法:50只SD大鼠依照随机对照方法分为5组,分别为假手术组(sham)、卒中组(MCAO)、卒中后抑郁组(PSD)、氟西汀治疗组(PSD+FLU)以及芬戈莫德治疗组(PSD+FTY)。采取大脑中动脉线栓法建立局灶性脑缺血模型,联合慢性不可预见的温和性应激(CUMS)及孤养法建立PSD模型。记录大鼠每周体重,运用敞箱实验和糖水偏好实验评估大鼠的行为学表现; HE染色观察大鼠海马组织形态; ELISA检测大鼠血清中IL-1β、IL-6和TNF-α的含量; Western Blot检测大鼠海马组织中IL-1β、IL-6和TNF-α的表达水平;流式细胞术检测大鼠外周血T细胞亚群水平。结果:与sham组和MCAO组相比,PSD组大鼠体重、敞箱实验得分及糖水偏好比例显著减低(P 0.05),HE染色显示海马组织CA1和CA3区呈现细胞萎缩和数量减少等现象,大鼠血清及海马组织中IL-1β、IL-6和TNF-α的表达水平均显著增高(P 0.05),大鼠外周血CD3~+T细胞和CD3~+CD4~+T细胞的数量升高(P0.05);与PSD组相比,PSD+FTY组大鼠体重、敞箱实验得分及糖水偏好比例显著升高(P 0.05); HE染色显示海马组织CA1和CA3区细胞损伤现象减少;大鼠血清及海马组织中IL-1β、IL-6和TNF-α的表达水平均显著降低(P 0.05);大鼠外周血CD3~+、CD3~+CD4~+和CD3~+CD8~+T细胞的数量均显著减低(P 0.05)。结论:FTY720能够有效改善海马组织损伤进而缓解PSD大鼠的抑郁行为,其机制可能是降低外周血T细胞数量从而进一步降低了血清及海马组织中IL-1β、IL-6与TNF-α水平。     

慢性轻度不可预见性应激对大鼠海马内甘丙肽表达的影响

目的:观察慢性轻度不可预见性应激(chronic unpredictable mild stress,CUMS)对大鼠海马甘丙肽(ga-lanin,Gal)蛋白及mRNA表达的影响,探讨Gal在大鼠实验性抑郁症发病过程中的作用机制。方法:将20只健康雄性SD大鼠,随机分为正常组和模型组,每组10只。孤养结合CUMS 21 d建造抑郁症模型,应用旷场试验和糖水偏好试验检测大鼠行为学变化,使用免疫荧光组织化学方法和逆转录多聚酶链反应(RT-PCR)法测定Gal及Gal mRNA的表达。结果:造模后模型组大鼠运动总路程、中央路程、周边路程及糖水摄入量均较正常组偏低(P<0.05);模型组与正常组相比,海马内Gal及Gal mRNA的表达同样降低(P<0.05)。结论:慢性轻度不可预见性应激可使大鼠海马内Gal及Gal mRNA的表达量下降,推测Gal在抑郁发病过程中可能起到神经保护作用。     

Secular change in menarche in women in Madrid

The age at menarche was estimated by recollection in 1617 women between the ages of 18 and 60 in Madrid and a nearby suburb, Pinto. The population of Pinto is working-class and the Madrid group, taken from residential neighbourhoods , belongs to the upper middle class. In both groups we found a diminution in average age at menarche, from 14.04 to 13.02 years in Madrid and from 14.55 to 13.16 years from about 1935 to about 1965 in Pinto. These changes have been more intense in the group which is less well-off economically, where living conditions have varied much more drastically.     

Intestinal helminthiasis in school children in Haiti in 2002

A survey on intestinal helminths in school children was conducted in Haiti in 2002. This first nationwide study involving the entire country was stratified by department according to urban and rural zones using the cluster method. Focusing on elementary school children (n=5792; age range 3 to 20 years), it involved 26 urban and 49 rural schools randomly selected. Stools were preserved in formalin and examined by the Ritchie technique. Thirty-four per cent of stools (1981/5792) tested positive for intestinal helminths with the following parasites identified: Ascaris lumbricoides (27.3%), Trichuris trichiura (7.3%), Necator americanus (3.8%), Hymenolepsis nana (2%), Taenia sp. (0.3%) and Strongyloides stercoralis (0.2%). The helminth prevalence was higher in rural (38.4%) compared to urban areas (30%). There was no significant difference in prevalence by sex and age. The importance of geohelminths changed from one department to another with the highest prevalence found in the Southern department of Grande Anse (73.7%) and the lowest prevalence in the Center department (20.6%). Five out of the country's nine departments had a similar prevalence varying from 25.5% to 28.2%. Intestinal helminthic polyparasitism was observed in a percentage of infested school children comprise between 3.4% and 28.6% according in relation to the geographical area. A program to fight against geohelminths in school children should be initiated as a public health priority. Albendazole is the drug of choice. Frequency of drug distribution should be based on the prevalence of geohelminths in each department.     

Experience in adult population in dengue outbreak in Delhi

A dengue outbreak has recently hit the Indian capital. We studied the clinical profile of adult patients. Five hundred and sixty patients of dengue infection were admitted in a specially created ward according to the criteria laid down by WHO. Haematemesis (28.28%), epistaxis (26.78%) and malena (14.28%) were some of the common presentations. Similarly lymphadenopathy, especially cervical (30.89%), palatal rashes (26.96%) and hepatomegaly (23.75%) were the most commonly encountered findings on physical examination. Most of the cases were of dengue fever with haemorrhage and only 2.5% cases were classified under dengue haemorrhagic fever or dengue shock syndrome. The average hospital stay was 3.4 days but only 9.8 hours in the eleven patients who died, suggesting their late arrival in preterminal situation giving little time for resuscitation. Thrombocytopenia was not a feature and only 12.85% patients had platelet count less than 70,000/cmm. Most of the patients who were admitted with thrombocytopenia, showed normalization in their platelet counts in next few days. Serological examination demonstrated evidence of recent dengue infection in 41.17% patients. Few patients required blood or platelet concentrate transfusion. Eleven patients died, three due to DIC, one of intracranial haemorrhage and seven due to massive gastric haemorrhage. Rest of the patients recovered completely. Thus we can conclude that recent outbreak in Delhi was of dengue fever with haemorrhage and mortality was very low in patients who came early to the hospital.     

Changes in the hypothalamic nuclei in in experimental allergy

Summary In rabbits subjected to prolonged sensitization and in which the Arthus phenomenon was induced there was a marked reaction of the hypothalamic nuclei. Staining by Gomori's method indicated a cellular swelling, loss of granules, and protoplasmic vacuolization in the supraoptic nucleus. There was a considerable increase in the size of the cross-sectional area of the cells. The same effects were much less well shown in the paraventricular nucleus. These results show that marked signs of increased neurosecretion developed in the animals at the height of the Arthus phenomenon.(Presented by Active Member AMN SSSR V. V. Parin) Translated from Byulleten Éksperimental'noi biologii i Meditsiny, Vol. 55, No. 4, pp. 110–113, April, 1963.     

Development in in vitro toxicology

There are three principal pressures driving the development of in vitro toxicology: (1) the need for more efficient testing systems to cope with the large number of xenobiotics currently being developed; (2) public pressure to reduce animal experimentation; and (3) a need for a better understanding of the mechanisms of toxicity. Within this, in vitro toxicology is focused on local, systemic, and target-organ toxicity. It is becoming increasingly apparent that a step or decision-tree approach using input of a variety of experimental data (physicochemical properties, biokinetics, cytotoxicity) provides the most efficient system for predicting toxicity. Examples of the use of in vitro toxicity systems for prediction of systemic toxicity and target-organ (liver) toxicity are presented.Originally presented at ECCP 93.