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Based on the detailed analysis of a variety of factors affecting stability of taxol production in Taxus cells, it was discovered that initial cell density and the relevant concentrations of substrates, elicitors and precursors significantly affected stability of taxol production; farther study showed that addition of elicitors at appropriate cellular state was the key to steadily improve taxol yield, especially, the proper cellular state was at the end of lag phase and at the beginning of the stationary phase, which was also proved by the cellular ultrastructure.

对影响红豆杉细胞紫杉醇产量稳定性的各类因素进行详细分析,发现细胞的接种密度以及与之相适应的培养基的基质浓度、诱导子浓度和前体浓度是影响紫杉醇产量稳定的重要因素;进一步研究发现,选择细胞处于合适的状态时加入诱导子是稳定提高紫杉醇产量的关键,细胞合适状态特别是延迟期末和稳定期初最合适,细胞状态的超微结构观察结果也证明了这一点;同时研究发现采用合适的同步化处理获得的细胞初始周期时相加入诱导子与紫杉醇产量稳定密切相关,且以低温同步化处理得到的细胞周期的分裂中期相的细胞加入诱导子不仅大大提高了紫杉醇的产量,而且诱导子加入时间大大缩短,有利于大大提高紫杉醇生产率。

Methods: The health adult human primary hepatocyte microsome was randomly divided into 12 groups for control and itraconazole with different concentrations in the range of clinical drug blood concentration, each group have 10 samples. Control groups were added culture fluid, itraconazole groups were added itraconazole with different concentration respectively, after cultured 30 minutes, substrates (phenacetin for CYP1A2, testosterone for CYP3A4) were added and cultured for another 40 minutes.

采用健康成人肝细胞微粒体,分为对照组和不同浓度的伊曲康唑组,共12组,每组10个样本,伊曲康唑组分别加入血药浓度范围内不同对应浓度的伊曲康唑,对照组仅加入培养液,孵育30 min后,再加入CYP450同工酶1A2和3A4的相应底物再孵育40 min。

Results indicated that the addition of these two fractions in atmospheric residue exhibited negative effect on the colloidal stability of residue system, and the negative effect of adding FCC slurry oil was stronger.

结果表明,加入两种油品后,体系的稳定性均呈下降趋势;不同油品对渣油胶体稳定性的影响程度不同,加入减四线抽出油后体系的稳定性比加入催化裂化油浆时高。

Result] The color and elasticity of steamed breads made from 250 g flour added with 50 g cactus juice and 30 g amomum juice were bad and their surface was not smooth;when the addition amount of cactus juice was 70 g,the elastic extensibility of steamed breads was bad;when the addition amount of cactus juice was 90 g,the elasticity of steamed breads was good and their surfaces were smooth.

结果] 在250 g面粉中加入50 g仙人掌汁、30 g砂仁汁蒸制的馒头色泽、弹性不佳,表面不光滑;加入70 g仙人掌汁时馒头的弹性延伸性不好;加入90 g仙人掌汁时,馒头的弹性佳,表面光滑。

When reached logarithm growth phase, it was poured or the medium was removed. 5 mL medium containing 10 mg/L mitocin-C was added , kept at 37 ℃, and then cultured in saturated humidity warm box with the CO2 of 0.05 fraction volume for 2 or 3 hours. It was coated with 0.1% gelatin in 6-hole plate, laid for over 30 minutes, and then threw away or the medium with mitocin-C was removed, washed with phosptat buffer for 5 times so as to remove the mitocin-C. 2 mL 0.05% trypsin digestive cells were added, and it was observed under microscope. When crevice appeared, cells became round (about 2-4 minutes), digestion was stopped by adding medium of the same volume, and blew up with straws repetitively to make it into monoplast suspension. Special-used cover glass was put in the center of cell counting chamber. Cells were sucked in by glass siphon, and cell suspension flew out at bucket of up or down-sides of counting chamber, until the cover glass was filled with fluid. Living cell were inoculated at concentrations of 3×108, 5×108, 1×109 L-1 after their number counted.

选取2~5代的小鼠胚胎成纤维细胞,待其至对数生长期倒掉或吸掉培养基,加入含10 mg/L丝裂霉素C的细胞全培养液5 mL,置37 ℃,体积分数0.05的CO2饱和湿度温箱中培养二三小时,在6孔板中加入0.1%明胶包被,放置30 min以上,倒掉或吸掉含丝裂霉素C的培养基,磷酸盐缓冲液清洗5遍,尽量除去丝裂霉素C,加入2 mL 0.05%的胰蛋白酶消化细胞,镜下观察,当细胞间出现裂隙,细胞变圆时(约2~4 min),立即加入等量的全培养液终止消化,并用吸管反复吹打,使之成为单细胞悬液,在细胞计数板中央放置专用的盖玻片,用玻璃虹吸管吸取细胞,让虹吸管在盖玻片上或下侧的计数板凹槽处流出悬液,至盖玻片下被液体充满为止。

Monocytes generated from bone marrow of Balb/cj mouse were cultured for 6 days in complete RPMI 1640 medium containing 10% FBS, rmGM-CSF and rmIL-4.50 mg·L-1 acteoside or isoacteoside was added to cells on day 6 of culture for 24 h.The surface molecules expression level of DCs and their phagocytose ability were analysis by flow cytometry.

采用细胞因子诱导法,从Balb/cj小鼠骨髓细胞贴壁分离获得单核细胞,加入含10%胎牛血清、10 μg·L-1重组小鼠粒细胞巨噬细胞集落刺激因子及重组小鼠白细胞介素-4(rmIL-4)的RPMI 1640完全培养基培养,在培养第6天,实验组加入车前子苯乙醇苷类化合物(10,50,100 mg·L-1),对照组加入RPMI 1640或LPS(1 mg·L-1),流式细胞仪检测DCs表面分子CD11c,CD86,MHC II和CD80的表达以及各组DCs吞噬功能的变化。

The survival rates of CEM cells cultured with cis-diamminedichicloroplatinum added 24 h later were higher than that cultured with hTERT ASODN and DDP added 24 h later. The survival rates of CEM cells cultured with DDP were similar with that cultured with hTERT SOND and DDP. In morphological observation of apoptotic cells using Giemsa staining, cells treated with DDP or DDP combined with hTERT ASODN ro SODN at 48 h, displayed classic apoptotic changes. Apoptosis rates of CEM cells treated with DDP for 48 h after 24 h of exposure to ASODN significantly increased. There was significant difference in the percentage of apoptotic cells of CEM cells between hTERT ASODN plus DDP and SODN plus DDP or DDP alone, respectively.

结果: hTERT ASODN作用于CEM细胞24 h再加入柔红霉素、长春新碱、足叶乙甙,对细胞生长的抑制分别与单用柔红霉素、长春新碱、足叶乙甙及hTERT正义寡核苷酸联合柔红霉素、长春新碱、足叶乙甙组相比,统计学上无显著差异(P>0.05)。hTERT反义核酸作用于CEM细胞24 h加入顺铂,再共同作用48 h,CEM活细胞均数为2.318×108 cells/L,与单用顺铂组(3.250×108 cells/L)及hTERT正义核酸联用顺铂组(3.175×108 cells/L)相比,对细胞抑制明显增强(P<0.05)。hTERT ASODN作用于CEM细胞24 h再加入顺铂作用48 h,细胞出现典型的凋亡形态学改变。hTERT ASODN与2.5 μmol/L顺铂联合作用于CEM细胞48 h的凋亡细胞百分率(19.47%)分别同SODN与顺铂联合作用组(6.97%)、单用顺铂作用组(6.02%)进行比较有显著差异(P<0.01)。

The processing technology of Huanghua pear fermented wine are : Huanghua pear →selection, washing →cleavage breakage→pretreatment→squeezing juice →pectinase clarification(pH3.5,1%pectinase solution adding 0.15%,45℃,50minutes)→dominant fermentation (1596 yeast, temperature 20℃,inoculation volume 7%,optimum initial pH3.1~3.4,volume loading 80%,sulfur dioxide 100mg/l,circle 8 days )→seperation and taking wine →after fermentation(10~15℃,14days)→seperation and disgorging→maturation→clarification (bentonite adding 0.16%)→filtration→adjustment→bottle

9黄花梨酒的加工工艺为:黄花梨→挑选、清洗→破碎→预处理→榨汁→果胶酶澄清(pH3.5,1%酶液的加入量为0.15%,温度45℃,时间50min)→主发酵(1596菌种,发酵温度20℃,接种量7%,最适初始pH3.1~3.4,装液量80%,二氧化硫加入量100mg/l,周期8天)→分离取酒→后发酵(10~15℃,时间14天)→分离除渣→陈酿→澄清(皂土加入量为0.16%)→过滤→调配→装瓶

METHODS: Normal human BMSCs were isolated and cultured by the whole bone marrow method. Cells of the third passage at 1×108/L were incubated on coverslip in a 6-well plate, and randomly divided into 3 groups: osteogenic induction group, which was added with primary medium and osteogenic inducer, supplemented with 10-8 mol/L dexamethasone, 10 mmol/L β-phosphoglycerol and 50 mg/L vitamin C; leptin osteogenic induction group was added with 50 nmol/l leptin in addition to osteogenic inducer. Cells in the blank control group were only treated with LG-DMEM containing 10% fetal bovine serum.

采用全骨髓法体外分离培养人骨髓间充质干细胞,传至第3代以1×108 L-1密度接种至预置盖玻片的6孔培养板中,设立3组:成骨诱导组添加原代培养液后,再加入含10-8 mol/L地塞米松、10 mmol/L β-磷酸甘油、50 mg/L维生素C的成骨诱导培养基;成骨+瘦素诱导组添加成骨诱导培养基后,再加入50 nmol/L瘦素;空白对照组仅加入含体积分数为10%胎牛血清的LG-DMEM培养液。

If you train yourself as you easily can, there are wonderful chances of usefulness before you: you can join the ranks of 15,000 Negro women teachers, of hundreds of nurses and physicians, of the growing number of clerks and stenographers , and above all of the host of homemakers.

假如你能培养自己,就像你现在可以轻而易举地办到一样,那么,在你的前头将出现奇妙的用武之地:你可以加入现在拥有一万五千名女性的教师队伍,加入成百上千的护士和医生队伍,加入人数愈来愈多的职员和速记员的队伍,而且最紧要的是,你还可以加入家庭主妇的行列。

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