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MATERIALS: Three Japanese big-eared rabbits with 3-months old were provided by Vital River Laboratory Animal Technology Co., Ltd., and the medical chitosan was purchased from Golden-shell Biochemical Co., Ltd.

材料:清洁级3月龄日本大耳兔3只,由北京实验动物技术有限公司提供。医用级壳聚糖为浙江金壳生物化学公司产品。

METHODS: BMSCs were obtained from Japanese big-eared rabbits, and in vitro cultured. Then the subcultured BMSCs were transfected by pCDNA3.1 plasmid, followed by incubation on swine SIS to construct the tissue-engineered skin. The growth of cells and phenotype of BMSCs were detected by flow cytometry.

日本大耳兔骨髓间充质干细胞进行体外培养扩增后,通过pCDNA3.1质粒将碱性成纤维细胞生长因子基因转染至生长状态良好的骨髓间充质干细胞,并将转染后的细胞接种于制备好的猪小肠黏膜下层上,进行体外联合培养,构建组织工程皮肤。

BMSCs were isolated, depurated, cultivanted, and identified,then incubated with the concentration of 25μg Fe per milliliter at 37℃in 5% CO2. The labeled cells were stained by Prussian blue/trypan blue,and observed under fluorescent microscope.2. The labeled cells of different density (1×104/ml,5×104/ml,1×105/ml,5×105/ml,1×106 /ml,5×106/ml)were imaged by MRI with T1WI, T2WI and T2*WI sequences;and the same density (5×104/ml,1×105/ml)labeled cells were imaged by T2*WI sequences at different time.Then the signal intensities were measured and statistically analyzed.3. The model of rabbit renal ischemia-reperfusion injury was set up and treated. Then BMSCs(5×105)were injected into 16 recipient rabbits(1abeled cells in 10,unlabeled cells in 6)from ear vein.MR images of kidneys were obtained respectively at the time points of 0,1,3,5, 8 days after transplantation and before transplantation. MR imaging findings were analyzed,which were correlated with histological findings.

实验方法1分离、纯化、培养、鉴定兔BMSCs并以SPIO以25μg Fe/ml培养液浓度标记,对标记后不同时间的细胞行普鲁士蓝染色和台盼蓝拒染后显微镜观察。2将不同细胞浓度标记细胞管(1×104/ml、5×104/ml、1×105/ml、5×105/ml、1×106/ml、5×106/ml),以不同扫描序列T1WI,T2WI,T2*WI(GRE进行MR成像,再选择相同细胞浓度组(5×104/ml、1×105/ml)进行不同时相MR成像,并测量信号强度,进行统计学分析。3缺血再灌注肾损伤模型建立和处理,然后将标记和未标记细胞(5×105个)经耳缘静脉移植入家兔体内(共16只:注入标记细胞者10只,注入未标记细胞者6只),两组均于注射前、注射后第0、1、3、5、8天应用MRI对移植细胞进行活体示踪并与肾脏组织切片对照,然后对收集的信号强度进行统计学分析。

Methods: Forty Japanese flap-eared rabbits were randomly divided into control group,model group,Chinese herb group,gene group and combined group.

日本大耳兔40只,随机分为对照组、模型组、中药组、基因组和综合组。

Totally 63 Japanese flap-eared rabbits, with birth age of 10-12 month, 33 females and 30 males, with body mass of 1.7-3.3 kg, were randomly selected.

随机选取10~12月龄日本大耳兔63只,雌33只,雄30只,体质量1.7~3.3 kg。

Methods: The Yifuqing granula was given to rabbit with febrilis caused by the TAB vaccine. The effects of the Yifuqing granula on the ache threshold value of Jimpy mice caused by Hot plate, on auricle inflammatory swelling of Jimpy mice caused by dimethyl benzene, on the bacterium of Jimpy mice in vivo, and on CPE caused by virus were observed.

观察一服清颗粒对三联菌苗所致兔发热的影响;采用小鼠热板法观察一服清颗粒对小鼠痛阈的影响;观察一服清颗粒对二甲苯所致小鼠耳廓炎性肿胀的影响;采用小鼠体内抑菌实验观察一服清颗粒对金葡菌感染小鼠死亡率的影响;采用细胞病变法观察一服清颗粒对病毒致细胞病变作用的影响。

Methods MSCs were cultured with inbred line small-ear cancellous bone of Ban Na pigs . A 1.5 centimeter segmental defect was created in the mid-upper part of the radial shaft of adult rabbit. The defects were implanted into composited xenogeneic bone in experimental group and xenogeneic bone alone in control group , the defects were not filled with anything in blank group . The repair capability of the defects was assessed by scan electron microscope before operation and physical , histology, X-ray , transmission electron microscope, SPECT and bone mineral density examinations 4 , 8 and 12 weeks after operation .

将MSCs与版纳近交系小耳猪松质骨在体外联合培养,兔桡骨中上段制成1.5cm的骨—骨膜缺损模型,实验组植入复合异种骨,对照组植入单纯异种骨,空白对照组不植入任何材料,分别于术前行标本的扫描电镜观察和术后4周,8周,12周各时间点行标本的大体观察,组织学观察,X线片观察,透射电镜观察,SPECT扫描及骨密度测试,比较其骨缺损区骨修复愈合情况。

In present study, we used the adult rabbits articular chondrocytes as the object of study, we researched the effect of TGF-β1 to promote chondrocytes proliferation and preserve the synthetise of special stroma in order to provide theortically and clinically basis of culture and proliferation in vitro of a-dult people articular chondrocytes.

材料与方法 1。成年兔关节软骨细胞分离、培养:取18月龄的成年日本大耳白兔1只,处死,片状切取关节软骨,解剖刀将软骨切成1.0mm~3大小碎块,消化,过滤,离心,收集关节软骨细胞,加入DMEM培养液(含20%胎牛血清、青霉素100U/ml,链霉素100U/ml),调整细胞密度为2.5×10~5/ml,置于37℃,5%CO_2孵育箱内培养。

METHODS: After anaesthetized, rabbits were cut at the skin for 2 cm long at the crossing of skull center and posterior canthus, skull was exposed and periosteum was separated, then a round skull window with diameter of 0.5 cm was drilled at 0.5 cm left or to sagittal suture and 0.5 cm posterior to coronal suture, after that, 35 g/L rose Bengal was slowly injected from ear-edge vein in dosage of 1 mL/kg by once.

兔麻醉后,沿颅骨的中央与眼后角垂直交叉处纵行切开皮肤约2 cm,暴露颅骨,刮离骨膜,在矢状缝左侧外0.5 cm与冠状缝后0.5 cm处,用钻头直径0.5 cm的颅钻钻透颅骨,造成颅骨圆形洞窗,随即由耳缘静脉按1 mL/kg一次性缓慢注入35g/L四氯四碘荧光素钠盐。约3 min后用冷光源(波长540 nm,功率140 lx)对准颅骨洞窗,连续照射8 min后,缝合皮肤切口。

Second, timid fear of panic, like the quiet courage of the Dutch rabbit small spaniel, suddenly heard the sound on the "shock market" phenomenon, loss of appetite would diminish.

二、胆小怕惊,喜欢安静荷兰垂耳兔的胆子很小,突然听到声响,就出现「惊场」现象,食欲会减退。

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