Cpne5蛋白在新生及成年小鼠各组织及其在不同胎龄胎鼠脑组织中的表达

目的:检测小鼠Cpne5蛋白在脑发育过程及不同组织中的表达水平,为Cpne5基因的功能研究提供线索。方法:提取新生、成年小鼠多种组织蛋白及11.5,12.5,13.5,14.5,15.5,17.5 d胎鼠脑组织蛋白,用免疫印迹法检测Cpne5蛋白表达量。结果:Cpne5蛋白表达于新生4 d小鼠的全部受检组织,大、小脑表达量最少,眼和肺的表达量高。成年小鼠检测结果相似,Cpne5蛋白的组织分布广泛,但脑组织含量低,表达量以卵巢、睾丸为高,其次为肺和心脏。胎脑的Cpne5蛋白表达量在胎鼠11.5、12.5 d较高,13.5 d降低,14.5至17.5 d再次升高,出生后又降低。结论:Cpne5蛋白在新生和成年小鼠广泛表达于各种受检组织,脑组织表达量最低;小鼠胎脑发育过程中Cpne5蛋白表达呈现先高后低,再升高再降低的变化趋势。     

面神经横断后生长抑素mRNA表达与c-jun mRNA表达的比较

正常脑组织中生长抑素(GIF)含量丰富,但在神经损伤后修复过程中GIF的作用还不清楚。以鼠面神经损伤后神经核内神经元为模型观察原位杂交GIF mRNA的变化。正常鼠脑GIF mRNA探针原位杂交阴性,GIF反义mRNA探针原位杂交可观察到阳性信号。实验中在假手术组面神经核内一直可测到GIF mRNA的表达。面神经横断组鼠。在手术侧面神经核内GIF mRNA的表达:第1天轻度下     

ADAM23在小鼠脑组织的表达

目的:观察整合素-金属蛋白水解酶23(ADAM23)在成年小鼠脑组织的表达及分布。方法:RT-PCR克隆ADAM23全长序列,体外逆转录合成地高辛标记的RNA探针;原位杂交法检测ADAM23在脑各组织的表达。结果:ADAM23 mRNA在小鼠大脑皮层区、海马CA3区、小脑的浦肯野细胞层(Purkinje cell)区、舌下神经核区高表达;而小脑、下丘的中央核、小脑核侧部区等弱表达。结论:ADAM23在成年小鼠脑组织的广泛表达尤其中枢神经系统的高表达,预示着ADAM23在中枢神经系统内有重要作用。     

小鼠诱导性多能干细胞移植治疗宫内缺氧新生鼠脑损伤

目的:研究小鼠诱导性多能干细胞(iPSCs)移植对宫内缺氧新生鼠脑损伤的治疗作用及机制。方法:于孕14 d开始孕鼠置于动物缺氧培养箱中,制作胎鼠宫内缺氧模型。孕鼠分娩后新生鼠经侧脑室注入BrdU标记的iPSCs悬液移植组和PBS缺氧组,对照组不治疗,H-E染色观察脑组织形态学变化、八臂迷宫测试动物记忆功能、免疫组织化学检测脑BrdU阳性细胞和神经颗粒素(Ng)和神经元烯醇化酶(NSE)的表达、原位杂交检测血管内皮生长因子mRNA(VEGF mRNA)的表达。结果:移植组小鼠的学习记忆能力和脑皮质区细胞结构较缺氧组均明显改善;脑切片检出BrdU阳性细胞存在,缺氧组Ng和NSE阳性细胞表达较对照组显著减少,而VEGF mRNA的表达较对照组显著增加;移植组Ng、NSE和VEGF mRNA表达均较缺氧组显著增加。结论:小鼠iPSCs移植于宫内缺氧的新生小鼠脑中,可减轻缺氧神经元的损伤和提高学习能力,其机制可能通过上调VEGF mRNA和Ng的表达有关。     

地高辛标记的大鼠nNOS mRNA探针的制备和应用

目的　制备大鼠神经元型一氧化氮合酶 (neuronalnitricoxidesynthase,nNOS)地高辛 (digoxigenin)标记的RNA探针 ,探讨nNOS在脊髓中的表达定位。方法　采用RT PCR方法 ,从大鼠脑组织中扩增nNOS基因mRNA部分片段 ,并经序列测定。以dig nNOSmRNA为探针 ,采用原位杂交观察成年大鼠脊髓组织中nNOSmRNA表达。结果　RT PCR法扩增出一特异产物与预期长度 2 4 0bp相符 ,T载体克隆测序与nNOS基因 10 0 %同源。原位杂交结果显示阳性信号出现在成年大鼠脊髓组织中。结论　采用RT PCR和T载体技术获得了大鼠脑组织nNOS基因克隆 ,dig nNOSmRNA探针原位杂交显示正常SD大鼠腰段脊髓组织中表达nNOSmRNA。     

脂多糖对不同发育期小鼠肝脏β-防御素3表达的影响

目的:观察在脂多糖(Lipopolysaccharide,LPS)诱导条件下,胎鼠(E14.5,E17.5),新生鼠(P0,P4)以及成年鼠肝脏中鼠β-防御素3(mouseβ-defensin 3,mBD-3)基因表达。方法:100只孕鼠随机平均分成2组:LPS感染组(E14.5、E17.5、P0、P4、a-dult),正常对照组(E14.5、E17.5、P0、P4、adult)(n=50)。LPS感染各组分别于E13.5,E16.5,E17.5(Embroynic day)天,孕鼠腹腔注射550μg.kg-1LPS(100μl),同时正常对照组也注射等量生理盐水,24 h后取出肝脏。提取RNA及蛋白,采用RT-PCR及Western blotting检测mBD-3的RNA及蛋白表达;同时取出左半肝经4%多聚甲醛固定后,石蜡包埋切片,免疫组化法检测mBD-3的阳性表达,图像分析系统进行定量分析,并进行统计学处理。结果:正常对照组小鼠肝脏在胎鼠以及新生鼠期均未检测到mBD-3的表达,而LPS感染组小鼠肝脏在胎鼠以及新生鼠时期均检测到大量mBD-3的阳性表达,且随着时间的增加呈上升的趋势(P0.05)。结论:mBD-3在正常的胎肝以及新生小鼠肝脏中未检测到表达,在LPS的诱导下,其在不同发育期小鼠肝脏组织中表达均明显增高。这提示不同发育阶段小鼠肝组织在受到外界感染侵袭后,mBD-3可能是肝组织抵抗微生物感染的重要抗菌分子。     

博尔纳病病毒感染对新生大鼠脑内单胺类受体基因转录的影响

目的分析博尔纳病病毒(BDV)感染对新生大鼠脑内单胺类受体基因转录的影响。方法选择病毒滴度为2·0×106FFU/ml的BDV病毒液对新生大鼠进行颅内接种,接种量为10μl/只新生大鼠。30d后用RT-PCR和间接免疫荧光方法确定BDV感染情况;并采用半定量RT-PCR方法检测BDV感染大鼠脑组织中多巴胺2(D2)受体和5-羟色胺2α(5-HT2α)受体基因的mRNA转录情况。结果病毒接种后新生大鼠的BDV感染阳性率为88·89%。病毒感染大鼠脑组织中5-HT2α受体和D2受体基因的mRNA转录水平均明显低于对照组,差异有统计学意义。结论BDV感染可以明显抑制新生大鼠脑组织中D2受体和5-HT2α受体基因的mRNA转录,提示单胺类受体基因的转录与表达参与BDV感染的致病过程。     

NDRG2在小鼠胚胎脑组织的表达特点

目的:观察NDRG2 mRNA在小鼠胚胎脑组织的表达与分布。方法:采用胚胎期小鼠E10.5 d、E12.5 d、E16.5d和E18.5 d进行脑切片的原位杂交。结果:NDRG2 m RNA在小鼠胚胎发育各期均有持续性表达,主要定位于端脑、中脑、菱脑、间脑、眼原基、脊髓、小脑原基、海马原基、丘脑、下丘脑和许多神经核团,神经细胞定位主要集中在细胞质中。结论:脑内NDRG2作用贯穿了整个脑的发育阶段,提示该基因在小鼠胚胎组织发育过程中有重要作用。     

大鼠脑组织中p75基因克隆及其探针制备

为了克隆大鼠p75基因并制备地高辛标记的p75 cDNA探针(dig-p75 cDNA),本研究采用RT-PCR法,从大鼠脑组织mRNA中扩增p75基因mRNA部分片段,克隆入T载体,并经序列测定。以dig-p75 cDNA为探针,采用原位杂交方法观察成年SD大鼠海马组织中p75 mRNA的表达。结果:RT-PCR法扩增出一种特异产物与预期长度386 bp相符,T载体克隆测序与p75基因100％同源。原位杂交结果显示,阳性信号出现在成年大鼠海马组织中。结论:采用RT-PCR和T载体技术获得了大鼠脑组织p75基因克隆,dig-p75 cDNA探针原位杂交显示正常SD大鼠海马组织中表达p75 mRNA。     

先天性HCMV感染胎鼠大脑皮层ET-1 mRNA的研究

目的 对先天性人巨细胞病毒（HCMV）感染的胎鼠大脑皮层内皮素－1（ET－1）mRNA进行测定,以探讨先天性HCVMV感染致脑损害的机制。方法 在建立先天性HCMV中枢神经系统（CNS）感染胎鼠模型的基础上,用逆转录－聚合酶链式反应（RT－PCR）测定受不同病毒剂量感染的胎鼠大脑皮层ET－1 mRNA,并用地高辛标记的ET－1寡核苷酸探针对大脑皮层细胞印片进行原位杂交以检测相应mRNA转录量及胞内定位。结果 在大脑皮层组织的上清液中HCMV分离阳性;病理学研究证实受染胎鼠大脑皮层表现为侵袭性脑膜脑炎性改变,并在神经细胞内发现特异性核内嗜碱性包涵体。RT－PCR和原位杂交研究发现,受染胎鼠大脑皮层内ET－1 mRNA转录量增加,以1．0ml和0．5ml组为显著,而0.25ml组与正常对照组比较无明显差别。结论 HCM可经胎盘垂直传播至胎鼠脑组织。先天性HCMV感染可刺激受染胎鼠CNS ET－1 mRNA的转录,且与母鼠所接种的病毒量存在一定的量效关系。这些结果提示,ET－1在先天性HCMV感染脑损害过程中,早期可导致组织缺血性改变,而晚期则与受损大脑皮层的功能恢复有关。这对了解先天性HCMV感染致NS损伤的机理将提供有价值的参考依据,同时也为临床防治和优生优育提供一种有价值的手段。     

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.