染色法

The expression of proliferating cell nuclear antigen in each group was detected by immuno-histochemistry method. The cell cycle distribution was measured by flow cyto- metic analysis.

MTT法及免疫组织化学染色增生细胞核抗原法检测细胞生长活性，流式细胞仪测定细胞周期时相变化。

TTC staining,RT-PCR and Western blot analysis were used to study the effects of muscone on infracted volume,EAAClmRNA and protein levels.

运用Western blot法和TTC染色观察缺血区EAAC1表达和梗塞体积；采用RT-PCR和Western blot法，测定海马EAAC1 mRNA和蛋白在缺血1h、6h、24h的变化。

MAIN OUTCOME MEASURES: The seeded cells were observed under inverted microscope; at 1, 2, 3 weeks after seeding, the BMSCs were treated with 4% paraformaldehyde and stained with hematoxylin-eosin; The protein content in seeded cells was determined by bicinchoninic acid assay, and the content of DNA was quantified using Hoechst33258 assay at 5, 10, 14 days after culture.

主要观察指标：倒置显微镜下观察细胞形态；于培养1，2，3周采用40 g/L多聚甲醛固定，常规组织切片，苏木精-伊红染色；在培养5，10，14 d用Hoechs33258荧光法定量测定细胞内DNA含量，BCA法测定蛋白质含量。

The surface antigen expressions of MNC were detected by flow cytometry. MNC were induced with DMEM medium, containing HGF alone, FGF4 alone, HGF+FGF4 or no growth factor, respectively. The markers of hepatic linear cells were examined before induction, and 7, 14, 21 and 28 d after induction by RT-PCR, immunocytochemistry and periodic acid-schiff in every group.

分别用肝细胞生长因子、成纤维细胞生长因子-4(FGF4)、HGF+FGF4、无生长因子4种处理因素进行诱导培养，于诱导前及诱导后的第7、14、21、28天采用RT-PCR检测AFP、白蛋白及C-met、FGFR2 mRNA的表达，免疫细胞化学法检测肝细胞抗原和CK18的表达，PAS 法进行糖原染色。

To produce the scientific evidence for developing and manufacturing new antitumor drugs.Methods: 1 The inhibitory effect on cell growth of Hela was measured by MTT assay in treated or untreated groups (3.125, 6.25, 12.5, 25, 50μg/ml TAM and control) for three different treatment times (24h, 48h and 72h).2 Apoptosis and cell cycle were measured by FCM in four experimental groups (0, 4, 16, 40μg/ml TAM) for 48h.3 Adopting Wright and Giemse's staining to observe the morphology of Hela cells which treated with 40μg/ml TAM.4 Using invasion experiment to detect the Hela cells'invasive abilities which treated with 40μg/ml TAM.5 The protein expressional levels of P-ERK, ERK, C-myc and Cyclin D1 in Hela cells untreated or treated with 4, 16, 40μg/ml TAM for 24h were measured by Western blot.6 Expression of anti-apoptotic gene bcl-2, apoptotic gene bax and MMP-9 in Hela cells of four experimental groups (0, 4, 16, 40μg/ml TAM for 24h), were observed by revers transcription PCR.7 The protein expression of P-ERK, ERK, Bcl-2 and Bax in Hela cells treated with 40μg/ml TAM for 24h observed by laser scanning microscopes.

1采用四甲基偶氮唑蓝法检测不同浓度北豆根总碱(3.125、6.25、12.5、25、50μg/ml)处理不同时间(24、48和72小时)对Hela细胞增殖反应的抑制作用。2采用流式细胞技术(flow cytometry，FCM)检测不同浓度北豆根总碱(0、4、16、40μg/ml)作用48小时，对Hela细胞凋亡及周期变化的影响。3瑞氏-姬姆萨染色后显微镜观察北豆根总碱(0、40μg/ml)作用24小时后Hela细胞形态学变化。4采用Transwell小室法检测北豆根总碱(0、40μg/ml)作用24小时后对Hela细胞侵袭性的影响。5采用免疫印迹方法检测不同浓度北豆根总碱(0、4、16、40μg/ml)作用24小时后，Hela细胞中磷酸化ERK、ERK、C-myc、CyclinD1的表达变化。6采用逆转录-聚合酶链反应(revers transcription PCR，RT-PCR)半定量检测北豆根总碱(0、4、16、40μg/ml)作用24小时，Hela细胞抗凋亡基因bcl-2、促凋亡基因bax、基质金属蛋白酶-9(MMP-9)的表达变化。7应用激光共聚焦显微镜(laser scanning microscope，LSM)观察北豆根总碱(0、40μg/ml)作用24小时后，Hela细胞内P-ERK、ERK、Bcl-2、Bax蛋白的表达变化。

Methods: Mice bearing in vivo transplanted tumor-H22 hepatic carcinoma were used to evaluate the drug"s activity of anti-tumor and GAPSGF"s efficacy enhancing effect by calculating the ratio of antitumor and q. HE dyeing was applied to observe the tumor"s morphological changes after the anti - tumor drug. It is observed the change of theperipheral white blood cells in serum and the body weight to analyze the GAPSGF"s toxicity reducing effect on the anti - tumor drug-Cytoxan. Applying ELISA method to observe the change of IL-2 in serum ,Applying carbon clearance method to test the function of uninuclear phagocyte,took the weight of the spleen and thoracic gland, analyzed the mechanism — efficacy enhancing and toxicity reducing of GAPSGF.

构建体内动物移植性H_(22)肝癌模型，通过计算抑瘤率与q值来分析药物的体内抗肿瘤活性，判断树舌多糖GF注射液的增效作用；采用瘤组织常规HE染色观察药物作用下的肿瘤细胞形态变化；观察外周血WBC数和体重改变来分析树舌多糖GF注射液对环磷酰胺抗肿瘤的减毒作用；采用ELISA法测定血清IL-2、小鼠碳粒廓清法测定单核吞噬细胞功能、取胸腺与脾脏称重测定脏器指数等以分析树舌多糖GF注射液增效减毒的作用机制。

Objective To explore the clinical value of the titer of serum HP antibody in patients with gastric diseases.Methods Serous HP antibody was detected with ELISA and HP was cultured,tested with urase,and stained with Gram method on smears which was observed microscopically and compared with proteus protein-enveloped test.Results ELISA showed high sensitivity and specificity.Conclusion Serous ELISA detection of HP antibody can be used widely in clinical diagnosis and epidemical screening without endoscopy and biopsy.

目的 探索胃部疾病患者血清中的幽门螺旋菌HP抗体滴度的临床价值方法应用酶联免疫吸附试验ELISA法检测HP抗体与HP培养，尿素酶试验和涂片革兰氏染色作镜下形态检查相比较，并同时与变形杆菌蛋白包被测定法作比较结果显示该法具有敏感特异性强的特点结论该法无需常规内镜检查，活检查HP而单纯用血清学检查，可广泛应用于临床诊断和流行病学人群普查

Methods The methods of Amsel, one step sialidase, amines and Gram-stain were performed simultaneously to diagnose bacterial vaginosis in 100 women.

随机选择就诊病人100例分别用Amsel法，组织多胺试验，一步法唾液酸酶活性的检测试验，革兰染色细菌评分法检测细菌性阴道病。

Leukocyte proteins concentrations were determined with Bradford Method. Phosphoproteins in the collected leukocyte proteins were enriched by BDTM Phosphoprotein Enrichment Kit. Finally the enriched phosphoproteins were separated by SDS-PAGE and dyed by argentation, and the electrophoresis map of SDS-PAGE was analyzed by Scion Image Tool.

刺激组加入表皮生长因子(40 μg/L)；抑制剂组加入细胞外信号调节激酶特异性抑止剂UO126 （10 μg/L）和表皮生长因子（ 40 μg/L）；对照组加入等量的培养基，用Bradford法测定细胞蛋白质含量，BDTM Phophoprotein Enrichment Kit分离富集白细胞磷酸化蛋白质，SDS-PAGE分离磷酸化蛋白质，银染法染色，采用Scion Image软件分析电泳结果。

MethodsThe cells of human gastric cancer cell line SGC7901 was treated respectively with Chrysin at different concentration 10、20、40、80μM for 48h. The proliferation inhibitory rate was measured by MTT assay. Acridine orangestaining and flow cytometry were used to detect apoptosis. Western blot assay was used to detect the expression of apoptosis related genes NF拨B,Bcl2,Bax.

方法分别用10、20、40、80μM的ChR处理人胃癌细胞株SGC7901 48h，采用MTT比色法检测ChR对SGC7901细胞的增殖抑制效应；采用丫啶橙染色、流式细胞术检测ChR诱导SGC7901细胞凋亡的发生；应用Westernblot法检测凋亡相关基因NF拨B、Bcl2、Bax的蛋白表达，并用计算机图像分析软件进行半定量分析。

I am accused of being overreligious," she said in her quiet, frank manner,"but that does not prevent me thinking the children very cruel who obstinately commit such suicide.""

客人们在卡罗利娜·埃凯家里，举止就文雅一些，因为卡罗利娜的母亲治家很严厉。

Designed by French fashion house Herm è s, this elegant uniform was manufactured in our home, Hong Kong, and was the first without a hat.

由著名品牌 Herm è s 设计，这件高贵的制服是香港本土制造，是我们第一套不配帽子的制服。