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The diluted monoplast suspension was equally assigned into 3 samples, and separately used for simple modified differential adherence, simple serum-free conditioned medium, and modified differential adherence + serum-free conditioned medium.

①无菌条件取大鼠嗅球组织,消化、离心去除上清液后,用含体积分数为0.2胎牛血清的DMEM/F-12培养基重悬,稀释的单细胞混悬液均分成3份,分别用于单纯改良差速贴、单纯无血清条件培养液纯化和改良差速贴+无血清条件培养液纯化的实验。

METHODS: Rat bilateral olfactory bulbs and olfactory mucosa at 1/3 nasal septum were obtained, sliced, digested in trypsin, and made into monoplast suspension. At 1×109/L, cells were incubated in uncoated 25 cm2 culture flask. At 18-20 hours, cell suspension was moved into another uncoated 25 cm2 culture flask (the first differential adhesion). At 24 hours, cell suspension was moved into a poly-D-lysine-coated 25 cm2 culture flask or poly-D-lysine-coated 6-well culture plate (the second differential adhesion). At 48 hours, parenchyma cells were removed after a half of medium was changed.

完整取大鼠双侧嗅球及剪取鼻中隔后1/3嗅黏膜,剪碎后胰酶消化,制成单细胞悬液,按1×109 L-1密度接种于未包被的25 cm2玻璃培养瓶中,18~20 h后将细胞悬液转移至另一未包被的25 cm 2玻璃培养瓶中(第1次差速贴),24 h后再将细胞悬液转移至经多聚右旋赖氨酸包被的25 cm2塑料培养瓶或经多聚右旋赖氨酸包被的6孔培养板中(第2次差速贴),接种培养48 h后半量换液以去除杂细胞。

METHODS: Olfactory bulb was sterilely collected from rats, digested, and centrifuged. After removal of the supernatant, the olfactory bulb was resuspended with DMEM/F-12 medium containing 0.2 volume fraction of fetal bovine serum. The diluted monoplast suspension was equally assigned into 3 samples, and separately used for simple modified differential adherence, simple serum-free conditioned medium, and modified differential adherence + serum-free conditioned medium.

①无菌条件取大鼠嗅球组织,消化、离心去除上清液后,用含体积分数为0.2胎牛血清的DMEM/F-12培养基重悬,稀释的单细胞混悬液均分成3份,分别用于单纯改良差速贴、单纯无血清条件培养液纯化和改良差速贴+无血清条件培养液纯化的实验。

In present study, the model of the low shear stress in carotid or hypertension was established by the ligation of partial distal branches of the left common carotid or the coarctation of aorta in SD rats respectively. In some rats the low shear stress in carotid was accompanied by the hypertension. The media and intimal thickness, as well as the ratio of vessel thickness to inner diameter were determined by morphometrical approach.

本文通过腹主动脉缩窄、结扎左颈总动脉的部分分支建立了高血压、左颈总动脉低切应力、以及高血压伴有低切应力大鼠动物模型;几何形态学方法观测左颈总动脉的厚及厚/内径比的变化;金属蛋白酶谱法分析MMP-2活性;免疫印迹法检测信号通路分子p-Akt、p-38分子以及Rho GDIα的表达变化。

Results: The results showed that:(1) The three types of NOS immunoreactivity were not found in rat testis until 30 days after birth;(2) NOS1 immunoreactivity was found in a few spermatocytes in rat testes on postnatal 30 days, and in spermatozoa present at the lumen surface of seminiferous tubules and some Leydig cells on postnatal 60 days;(3) NOS2 immunoreactivity appeared in a few spermatocytes, Sertoli cells and peritubular myoid cells on postnatal 30 days. On postnatal 60 days, NOS2 immunoreactivity appeared in some Leydig cells, peritubular myoid cells, Sertoli cells, very few spermatocytes and the head of immature spermatozoa in some seminiferous tubules;(4) NOS3 immunoreactivity was detected in a few spermatocytes on postnatal 30 days, as well as in the smooth muscle cells of blood vessels on postnatal 30 days and 60 days.

结果:(1)生后4、7、14 d大鼠睾丸未见3种NOS免疫阳性反应;(2)生后30 d少数精母细胞及生后60 d生精小管腔面精子和间质细胞呈NOS1阳性;(3)生后30 d少数精母细胞、支持细胞和管周类肌细胞呈NOS2阳性,而生后60 d NOS2阳性反应见于睾丸间质细胞、管周类肌细胞、支持细胞、极少数精母细胞和不成熟精子头部;(4)生后30 d睾丸内少数精母细胞和血管呈NOS3阳性,生后60 d NOS3阳性反应仅见于血管

Although heart wall thickness was reduced overall, specific areas exhibited morphological changes consistent with myxomatous degeneration in the walls of knockout hearts.

尽管整体心室厚度下降,但是特定区域的形态学改变和基因敲除小鼠的心室粘液瘤的形成一致。

Methods Human umbilical cord blood mononuclear cells were isolated by Ficoll density-gradient centrifugation. EGM-2 culture fluid was added, and then cells were plated on dishes coated with human fibronectin. After 48 h, the nonadherent cells were collected and replated on fibronectin-coated dishes.

采用Ficoll密度梯度离心法从人脐血中分离单个核细胞,加入EGM-2培养液,接种于包被人纤连蛋白的培养皿中贴培养,收集48h后的悬浮细胞重新贴培养至第7天,利用免疫荧光和流式细胞术鉴定。

METHODS: mononuclear cells were isolated from axillary draining lymph node, and the adherent cells were cultured with rhGMCSF and rhIL4 to induce DC. The nonadherent cells were cultured with IL2 to induce tumor draining lymph node cells.

以多种细胞因子联合诱导腋下淋巴结单个核细胞中的贴细胞为DC,非贴细胞与IL2共培养后诱导成为肿瘤区域引流淋巴结细胞,用自体乳腺癌细胞冻融抗原刺激DC,并和TDLNC共培养,以诱导肿瘤抗原特异性CTL。

METHODS: The mononuclear cells were isolated from axillary draining lymph node, and the adherent cells were cultured with rhGMCSF and rhIL4 to induce DC. The nonadherent cells were cultured with IL2 to induce tumor draining lymph node cells.

以多种细胞因子联合诱导腋下淋巴结单个核细胞中的贴细胞为DC,非贴细胞与IL2共培养后诱导成为肿瘤区域引流淋巴结细胞,用自体乳腺癌细胞冻融抗原刺激DC,并和TDLNC共培养,以诱导肿瘤抗原特异性CTL。

METHODS: Bone marrow mononuclear cells were isolated from 5M4/M5 leukemia patients with high CD14 expression, and then divided into 3groups: adherent leukemia cells, nonadherent blasts, and total unfractioned blasts.

取5例初诊CD14高表达的M4或M5型白血病患者的骨髓标本,分离单个核细胞(bone marrow mononuclearcells,BMMNCs),将白血病细胞分为3组:贴白血病细胞组、非贴白血病细胞组及总白血病细胞组。

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