细胞计数

After cultured for 4 days,the cells were stained with TRAP. Multinucleated(≥ 3 nuclei) TRAP-positive cells were counted as osteoclasts, and the expression of TRAP mRNA were determined by RT-PCR.

2.0？10.0？50.0 ？滋g/ml，对获得的各组破骨细胞行抗酒石酸酸性磷酸酶染色计数阳性破骨细胞(核≥ 3)的数目；提取总RNA用逆转录-聚合酶链反应检测TRAP mRNA的表达？

Methods The neuroepithelial stem cells were isolated and cultured by microdissection, mechanical blowing and serum-free suspension culture.

方法采用显微解剖、机械吹打、无血清悬浮培养方法分离培养神经上皮干细胞，采用巢蛋白免疫细胞化学染色技术检测神经上皮干细胞，用NSE和GFAP免疫组化染色检测并计数神经细胞和神经胶质细胞。

Methods: The human KDR promoter (-225 bp~+127 bp) was cloned by PCR. Subsequently, a recombinant adenoviral plasmid carrying KDR promoter for the HSV-tk gene was constructed with the AdEasy system. As a control, the plasmid pAdCMV-tk carrying CMV promoter for the HSV-tk gene was also constructed. 293 packaging cells were transfected by both newly-constructed plasmids and the infectious viruses AdKDR-tk and AdCMV-tk were generated. Then the KDR-producing cells and the KDR nonproducing cells (HepG2) were infected with AdKDR-tkand AdCMV-tk, followed by ganciclovir administration.

应用PCR克隆出人KDR基因启动子序列－225bp~+127bp，以AdEasy system为载体，构建携带受KDR启动子或CMV启动子调控tk基因表达的2种重组腺病毒质粒pAdKDR-tk和pAdCMV-tk，在293细胞中包装、扩增后，体外感染表达KDR的脐静脉血管内皮细胞系HUVEC和不表达KDR的肝癌细胞系HepG2，并给予不同浓度的丙氧鸟苷处理，5d后收集存活细胞并计数。

Nuclear translocation of protein PDX-1 in different groups: Indirect immunofluorescence showed PDX-1 was mainly located in cytosolic slightly in the basical station, while translocated to nucleoplasmic greatly followed by stimulated with 16.7mM glucose.

为量化这种核转位程度，每组计数100个细胞并分别记数有荧光信号在核内表达的细胞数N及荧光信号在核周围表达的细胞核数C，比较各组的N／C值。

One has to search and count mitotic figures carefully in better preserved areas, and look for the other atypical features - hypercellularity, small cells with high nucleocytoplasmic ratios, patternless growth, and brain parenchymal invasion.

应该在保存好的区域仔细寻找计数核分裂，寻找其它非典型特征：细胞密集、核浆比高的小细胞、无序生长方式、脑实质浸润。

Methods: A brain concussion was produced in rats using a simple pendulum device model. One control (n=12) and six experimental groups were used(n=12 in each group). The cerebral concussion groups were assigned to sacrifice times ranging from 1 day to 24days after injury.A Morris Water Maze memory paradigm was used to assess learning and memory function. Pycnosis degeneration or necrosis neurons in the locus cerebral cortex, dorsal hippocampus, dentate fornix and brainstem reticular formation were observed and counted after brain concussion. Cholinergic neurons in the basal forebrain and brainstem reticular formation were identified and quantitated using choline acetyltransferase.

应用自制单摆式闭合性机械打击装置建立脑震荡大鼠模型；成年SD大鼠随机分为1个对照组(n=12)和6个脑震荡组(即伤后1、2、4、8、16、24d，n=12)；应用Morris水迷宫(Morris Water Maze，MWM)实验评价大鼠的学习记忆功能；用光镜对大鼠的上述脑区的固缩变性坏死细胞进行观察和计数；用免疫组化方法检测基底前脑和脑干网状结构的胆碱乙酰转移酶(cholin acetyltransferase，ChAT)阳性细胞的表达。

Get the neonate Sprague-Dawley rats medulla spinalis, through the conventional primary cell culture procedure, we use the method of difference adherence time and get the motor neurons of anterior spinal cord. When the neuron is in maturity, we make the mechanical injury model in vitro. All the models were divided into four groups: group A is control group; group B is 100μM ATP group; group C is 100μM ATP+20μg/ml suramin group and group D is 100μM ATP+10μM ouabain+10μg/ml Thapsigargin group. Culture the four groups neurons, after one day, we count the motor neuron, observe the survival and activity of neurons through MTT shade selection experiment, use flow cytometry to analyze the percentage of apoptosis of motor neurons of anterior spinal cord and detect the expression of protein p-GSK-3β(ser9) through Western-Blot technology.

方法取新生大鼠脊髓，通过常规的原代细胞培养程序，采用差速贴壁法分离出大鼠脊髓前角运动神经元，培养成熟后制作神经元机械损伤体外模型，分为A组：对照组、B组：100μMATP组、C组：100gMATP+20μg/ml Suramin组和D组：100μM ATP+IOμM Ouabain+10μg/ml Thapsigargin组，对各组机械损伤的运动神经元进行培养，1天后分别进行运动神经元计数、MTT比色实验观察运动神经元的存活及活性、流式细胞仪分析机械损伤的脊髓前角运动神经元凋亡百分率和Werstern Blot技术检测p-GSK-3β(ser9)的表达。

5Ug/ml without toxical effect on both cells activity was selected as the high dose to observed the effect on HUVEC and human peripheral polymorphonuclear cell activity by MTT and trypan-blue assay repectively.

2.2 疡愈涂剂对白细胞与血管内皮细胞粘附影响的分子机制用MTT法检测YYTJ对HUVEC活力的影响，用台盼蓝计数法检测YYTJ对人外周血中性粒细胞活力的影响。

Trypan blue exclusion was used to count the alive cells; morphological changes were...

通过台盼蓝拒染法计数活细胞；观察ATRA干预前后细胞形态学变化；RT PCR分析TrkA表达情况。

Results The reproductive activity of the bone marrow ceils irradiated by 6 and 8 cGy could be improved significantly in vitro.

用低剂量照射的骨髓细胞进行骨髓移植后，受体小鼠骨髓单个核细胞数和外周血细胞计数普遍高于相应的对照组。

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务，乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐，因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。