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Cumulus-enclosed oocytes from follicles in prepubertal gilt ovaries from alocal abattoir were cultured in modified TCM199 with and without 10 IU/ml PMSGand hCG for 22 hours respectively.After fixation and dying in 1% acid orcein,itshowed that 77.6% oocytes reached MII stage.Maturated oocytes were denudedwith 1mg/ml hyaluronidase,and fertilized with fresh or thawed sperms in modifiedTBM for 6 hours,then cultured in NCSU23 for 14 hours.The fertilization rates were87.5% for fresh sperms,64.1% for thawed sperms.

抽取从屠宰场获取的猪卵巢中的卵母细胞，在加入和不加hCG和PMSG的mTCMl99培养液中分别培养22小时，地衣红染色发现77.6％的卵母细胞达到MII期；去卵丘细胞后在mTBM中用鲜精或冻精解冻后受精6小时，在NCSU23中培养14小时后固定染色发现受精率分别为87.5％和64.1％；电激活后在NCSU23中培养20小时后固定染色发现原核形成率为94.1％。

The immunocytochemistry was used to identify the RASMC.RESULTS: The RASMC were successfully cultured by the adherent method of tissue explants and grown in the "peak- valley" mode. The cultured RASMC were fusiform shape with a big oviform or round nuclei rich in cytoplasm. Immunohistochemical method ( S-P method) showed strong expression of a- smooth muscle actin.

结果：组织块贴壁法成功培养出SD大鼠胸主动脉平滑肌细胞，培养的细胞呈典型&峰-谷&样生长，HE染色细胞呈梭形，胞浆丰富，核大而圆或椭圆，免疫组化S-P法检测a-平滑肌肌动蛋白单克隆抗体呈强阳性表达。

Methods: Between October 1999 and March 2001, 20 patients with severe acute pancreatitis following peripancreatic or peritoneal fluid collection were observed. Blood and fluid collection were dynamically cultured, at the same time, microorganisms cultured were identified, and then drug sensitivity was performed.

对1999年10月至2001年3月间收治的重症胰腺炎伴胰周或腹腔积液的20例患者进行动态血液、腹腔积液细菌培养、鉴定及药物敏感试验。

Cultured for 48 hours, the floating cells were removed and the adherent cells were continued to culture with cytokines including IL-4 and GM-CSF. After 14 days culture, the induced DCs were observed by photics microscope and electron microscope, also co-cultured with native T cells derived from spleen, its stimulating function was detected by MTT assay.

经过光镜，透射电镜观察培养DCs的形态学特征；通过同种T细胞混合培养，采用MTT比色分析法测定不同浓度的DCs发同种T细胞增殖的能力。

Results The strongest photoinactivation effect occured in the HEP-2 cells co-cultured with 1 mmol/L ALA for 8 h. The killing activity of ALA-PDT to the HEP-2 cells was positively correlated with the power density of the irradiation (r=0.88, P＜0.05).Conclusions ALA-PDT can effectively kill the cultured HEP-2 cells in vitro. The optimal killing activity was related to the time and the power density of the laser irradiation.

结果 (1)光照时机选择在ALA与HEP-2细胞培养8 h时杀伤效应最强；(2)ALA-PDT对HEP-2细胞的杀伤效应与照光能量呈正相关r=0.88，P 结论 ALA-PDT能有效地杀伤体外培养的肿瘤细胞，其杀伤最佳效果与照光时机和照光能量有关。

Methods Acanthamoeba polyphaga and Legionella pneumophila were co-cultured under laboratory condition. At consecutive time points during the culture, smears of the cultured products were made on glass slides for staining purposes. Different types of stainings including Gram′s staining, Gimenez staining, Giemsa staining and immunofluorescence were used to determine the best method for the identification of amoebal pathogens.

方法嗜肺军团菌与多噬棘阿米巴共培养，取不同时点的共培养物制作涂片，采用革兰氏染色、吉曼尼兹染色、姬姆萨染色、免疫荧光染色等多种方法，用光学显微镜及荧光显微镜观察鉴别阿米巴滋养体与其胞内嗜肺军团菌，并比较这些染色方法的效果。

The results showed that, using transient GUS expression, the frequency of resistant calli and shoot differentiation as a criteria, wheat mature embryos were cultured on the preculture medium of CM4C1P for 14 days, and immersed in inoculation suspension Ⅰ for 3 hours, then the explants were co-cultured under desiccation conditions for 3 days, which was optimal conditions for Agrobacterium-mediated transformation of wheat mature embryos.

为了进一步完善根癌农杆菌介导的小麦遗传转化体系，以小麦成熟胚为转化受体，研究了根癌农杆菌介导法转化小麦的主要影响因素，结果表明，以gus基因瞬时表达率、抗性愈伤组织获得率和分化率为转化指标，小麦成熟胚在预培养基CM4C1P上预培养14d后，用侵染培养液Ⅰ侵染3h，再经干燥共培养方式处理3d，是根癌农杆菌介导成熟胚转化的合适条件。

AIM: To observe the effect of diterpenoid compound 5F of Pteris semipinnate L on apoptosis in cultured human pterygium body fibroblasts. METHODS: Fibroblasts collected from human pterygium body were cultured in vitro with 5F at different time point.

目的：观察中药半边旗二萜类化合提取物5F对体外培养的人翼状胬肉成纤维细胞周期的影响及5F诱导细胞凋亡过程超微结构的变化，初步探讨5F对HPFs作用的机制。

Methods Pterygial samples were extracted and collected and the pterygial endothelial cells and pterygial fibroblasts were cultured alone, conditional and co-cultured to form different culture systems. The methods included that to select the suitable intensity of ultraviolet by MIT, to detect the changes of curves of growth about two kinds of cells by MIT and to explore the developments of protein and RNA of vascular endothelial growth factor and fibroblast growth factor-basic in three culture systems under ultraviolet whose intensity is 20 mJ/cm^2 by ELISA and RT-PCR.

收集翼状胬肉标本，采用血管内皮细胞和成纤维细胞单独培养、条件培养和共同培养的方法构建体系，用MTT法选择紫外线照射细胞的适宜强度；采用MTT法绘制强度20mJ/平方公分紫外线照射下细胞生长曲线；采用ELISA和RT-PCR检测紫外线照射下3种体系中细胞上清液和细胞中血管内皮细胞生长因子和成纤维细胞生长因子的蛋白和RNA含量变化。

①Rat MSC and VSMC were cultured and identified, respectively. MSC were labeled with DAPI firstly, and then co-cultivated with VSMC. The changes of morphology and ultrastructure of co-cultured cells were observed. Immunfluorescence analysis was performed by using monoclonal antibodies against specific antigen.②We established the regulatable system in two steps: a stable MSC line expressing rtTA has been constructed and characterized firstly by transfected with pUHD 17-1hyg and then selected by hygromycin B; in a second step, this line was used for trandfer the AT2R gene to MSC to get the well establishing double stable MSC lines;③The expression of AT2R regulated by doxycycline was evaluated by western blot;④The MSCs were transduced into rat carotid arteries with regulatable AT2R gene after the establishment of rat carotid balloon injury restenosis model. The intimal/medial area ratio were measured by digital analysis system.

研究方法：(1)密度梯度离心法及胶原酶消化法分别培养原代大鼠MSC及VSMC，细胞共培养并行免疫荧光化学染色和透射电镜观察超微结构；(2)组成受Dox调控的哺乳动物表达系统的四种成分的转化、扩增及提纯并酶切鉴定；(3)采用常规分子生物学方法连续两个回合转染体外培养的MSC，并分别采用发光计检测不同细胞克隆萤光素酶活性改变以及RT-PCR方法检测AT2R目的基因mRNA表达情况，根据各个细胞克隆受Dox调控表达的程度，选择低背景、高诱导表达AT2R的细胞系，作为双重稳定MSC细胞系；蛋白免疫印迹法观察该细胞系在Dox调控下AT2R表达的时相性、持续性及在不同浓度Dox调控下的表达情况；(4)建立大鼠颈动脉球囊损伤动物模型，将双重稳定MSC在术中导入血管，分别于14 d、28 d进行病理切片，检测可调控AT2R对新生内膜增生的影响；采用RT-PCR免疫组织化学免疫荧光等技术观察AT2R基因在新生内膜中的表达以及细胞外基质成分表达的改变，TUNEL法检测血管组织中细胞凋亡的变化情况。

Means the parent directory, so this command means to execute "toolchain.sh," which is in the current directory.

代表父目录，所以这个命令就是执行当前目录下的"toolchain.sh"。

Yes,In fact,I'm on our city ream.

是的。事实上我是我们市队的。