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After placement in a stereotaxic apparatus, A microdialysis probe was implanted in the left hippocampal CA1 region (2 mm to the left and 3.6 mm posterior to the bregma and 2 mm below the cortical surface).

切开头皮,用微型颅钻分别在左右海马CA1区(前囟3.6mm和中线左、右旁开3mm)打开一直径约2mm的圆孔,将微透析探针刺入左侧海马CA1区2mm,用乳酸钠林格氏液以2.5μl/min的速度持续灌注,稳定60min待脑电图波形稳定后收集脑组织透析液,间隔时间为10min,直至再灌注后2h。

RESULTSdiabetic rats showed typical symptoms involving elevated serum glucose and weight loss (2)The rats in DM+VaD group reacted slower and made more error numbers in Y-maze than the rats in VaD group did, with a prolonged total reacting time in a whole day.(3)More neurons with serious damages, such as edema, ischemia or pyknosis could be found in DM+VaD group than in VaD group. More disarrangement of cone neurons, obvious glia proliferation, and more neuron apoptosis were found in CA area of hippocampus. CONCLUSIONS (1)2-VO in STZ-induced diabetic rats succeeded in establishing VaD animal models.(2) The cognitive dysfunction induced by VaD rats had been deteriorated by diabetes mellitus.(3)Morphology evidences proved that diabetes mellitus aggravated brain damages induced by VaD.PART II Effect of diabetes mellitus on the cholinergic nervous systemin CA1 area of hippocampus of VaDOBJECTIVE To estimate the role of cholinergic nervous system in hippocampus during the process of cognitive dysfunction aggravated by diabetes mellitus in DM+VaD group. METHODS Observe the changes of ChAT protein by immunohistochemistry stain. Chose three rats randomly from each group at each pointat two week, four week, and eight week, then sepa

经腹腔注射链脲佐菌素诱导慢性实验性糖尿病,1周后永久性结扎双侧颈总动脉(2-VO)制作VaD模型记录术后2周、4周和8周各组大鼠的体重及血糖;应用Y-迷宫检测空间定向学第二军医大学博士论文中…丈摘…要1ia;一色恤~﹁染澎糕行HE染色观察组织病理学孪叱;胶质细胞原纤维酸性蛋白protellZ,Cf冰尸标记观察星形胶质细舱的激活情况、橄声软公点娜育票票众糯黑橄居端粗默孺篇筑价井袱第二部分糖尿病对血管性痴呆大鼠海马c心区胆碱能神经系统的影响探讨海马CAI区胆碱能神经系统在糖尿病加重确D一大鼠认知障碍中所实验动物及分组同第一部分,应用免疫组织化学染色方法现察海马CAI{介狱牛第晕军l袭拔常博粗{论义

RT-PCR/Southern blot was used to detect the change of Brn-4 mRNA expression in hippocampus after fimbria fornix transection.

取各组大鼠的海马组织,提取总RNA,采用RT-PCR/Southern 杂交方法分析穹窿海马伞切割后海马内Brn-4 mRNA的表达变化。

METHODS: The normal hippocampi and hippocampi on the 14th day after the hippocampal fimbria transection were prepared into homogenate used for 10% native- polyacrylamide gel electrophoresis, and the differential proteins of 83 ku were electroeluted. The protein concentration was adjusted to 300 mg/L.

取6只正常大鼠及6只切割海马伞后14 d大鼠的海马组织制成匀浆,进行非变性聚丙烯酰胺凝胶电泳,根据染色结果切取含有83 ku差异蛋白条带进行电洗脱,定量后调整蛋白浓度为300 mg/L。

Then hippocampi were isolated and total RNA was extracted. RT-PCR/Southern blot was used to detect the change of Brn-4 mRNA expression in hippocampus after fimbria fornix transection.

取各组大鼠的海马组织,提取总RNA,采用RT-PCR/Southern杂交方法分析穹窿海马伞切割后海马内Brn-4 mRNA的表达变化。

IL-1βmRNA expression in the developmental brain in Wistar rat: IL-1β mRNA in hippocampus, cerebral cortex and subcortex was detected by postnatal day 1 and gradually increased with development, reached a peak at postnatal day 21, and then gradually decreased to lower level at adult; In P77PMC rat, IL-1β mRNA expression was similar to Wistar rat before postnatal day 14, then gradually increased and surpassed Wistar rat at postnatal day 28, reached a peak in adult.

原位杂交显示谷氨酸诱导Wistar大鼠惊厥和P77PMC大鼠听源性惊厥后脑内IL-1βmRNA表达情况。结果发现,两种不同惊厥模型大鼠惊厥后脑内IL-1βmRNA表达存在明显的差异。谷氨酸引起惊厥诱导脑内皮层、海马和丘脑IL-1βmRNA表达增高的程度远较P77PMC大鼠听源性惊厥为低,而且惊厥后IL-1βmRNA表达增加较慢,于4-8h达高峰,而听源性惊厥可于惊厥后2h迅速达高峰。但两种形式惊厥诱导脑内IL-1βmRNA表达最高的部位均是海马。

Results: Remarkable pathological changes were found in h ippocampus: Nissl bodies decreased or disappeared, cyton swelled. The water cont ents and the level of LDH and NO were increased in scalded rats. CNTF could rema rkably improve the pathological changes and reduce the increase of LDH, NO a nd the water contents.

结果:大鼠严重烫伤后,脑组织含水量增加,海马出现明显的病理变化,尼氏小体减少或消失,胞体肿胀,LDH和NO的含量增加;给予CNTF后,脑水肿、海马的病理变化有明显改善,并能降低 LDH和NO的含量。

The steps are as the following:1、Preparing the cerebrospinal fluid one hour before the experiment,take some of them for cryopreservation;2、Without paralyse,cuting down the rat head quickly and dislodge the hippocamp part which is used in the experment in freezing liquid,and trim the hippocamp distrct.3、Fixing the hippocamp tissue on the vibratome,to cut down a brain slice with 300μm,use the haustorial tube to move the slice to the preincubate dish,cultivanting it for one hour.Then move it to the record incubation chamfer.4、Preparing the glass micloelectrode and fill it with NaCl,the corcentation is 3mol/L.5、adjusting the recorder,after cultivate the brain slice in the incubation chamfer for 2 hours,the expermentize can be gone on.6、In order to ensure the accurate of the experimental result,use only one medicine concentration for one brain slice in every experiment.

本实验研究方法是:1、实验前1小时配制好所需脑脊液并充以95% O2和5% CO2的混合气,取小部分冷冻保存;2、在乙醚麻醉下,快速断头并在冷冻液里取出包含海马的大脑部位,修整出所需海马区;3、将海马组织固定于振动切片机上切出300μm厚的海马脑片,用广口吸管转移至预孵育皿培养一小时,后再转移到记录用孵育槽内;4、拉制好玻璃微电极,并充以3mol/L的NaCl;5、调试好记录仪器,将脑片在孵育槽内培养2小时后进行实验;6、为了保证实验结果的准确性,每一块脑薄片只进行一个浓度的药品实验。

In 1-7d group, the difference between experimental group and control group was not significant. In 14-21d group, Bcl-2mRNA and P53mRNA were expressed lightly. In 28d group, the expression were comparatively strong. The results indicated that, after exposure to infrasound with some parameters, with the days elapsing, Bcl-2mRNA expression in neurons of mice hippocampus displayed certain regularity. The progressive increase suggested that infrasound could injure hippocampus, and resultantly, injure and repair neurons DNA.

据此,我们认为:当一定参数的次声作用小鼠后,随着作用天数的延长,小鼠海马内神经细胞Bcl-2mRNA表达有一定规律性,总体趋势表达增强,随次声作用次数进一步增多,21d组和28d组有产生适应的倾向,其意义在于次声作用后引起海马损伤,使神经元DNA损伤并产生修复。

ResultsThere was no expression of Caspase-3 in group A. The expression of Caspase-3 in group C was decreased compared with that in group B(P<0.01. In groups B and C, expansion and vacuolization of mitochondria, swollen neurons and apoptotic bodies appeared, which suggested that WBH could induce neuron injury, and the neuron injury in group C was more severe than that in group B.

结果A组大鼠海马区神经元无Caspase-3表达,B、C两组大鼠海马区神经元有Caspase-3表达,B组较C组Caspase-3表达平均灰度值减小。B、C两组大鼠海马神经元超微结构发生改变、细胞器水肿,部分线粒体空化,内质网稀疏,突触前膜、后膜结构不完整,可见自噬体结构;B组大鼠的神经元超微结构损害较C组轻。

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