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1 Day after injury ruptured capillary could be seen in ganglion cell layer, 4 weeks after injury cells in each layer arranged sparsely and disorderedly, in some RGCs chromatin became dense, 8 weeks after injury the cells in each layer became fewer and large amount of RGCs without nucleus could be seen.

3光镜下伤后1 d视网膜神经节细胞层出血,伤后4周视网膜各层细胞稀疏、排列欠整齐,GCL散在核染色质浓集、边聚的节细胞,伤后8周视网膜各层细胞明显减少,GCL内大量空化节细胞。

BALB/c mice were co-inoculated muscularly by this plasmid and the nucleic acid vaccine plasmid pVAXGE expressing the HIV-1 (Human immunodeficiency virus type I) gag-gp120 protein. The level of serum antibodies after immunization showed that the specific antibodies against HIV-1 appeared at the second week and raised to the peak level at the sixth week for the co-immunization group.

将它与表达I型人免疫缺陷病毒(Human immunodeficiency virus 1, HIV-1) gag-gp120的核酸疫苗质粒pVAXGE共同肌肉注射BALB/c小鼠,免疫3次后,以ELISA法检测免疫小鼠血清中抗HIV-1抗体水平,结果显示联合免疫组小鼠在免疫2周后已有抗体产生,6周后进入高峰。

In order to develop a safe and effective immunoadjuvant to enhance the immunity and resistance of animals against infection, a novel CpG Oligodeoxynucleotides containing 11 CpG motifs was synthesized and inserted into the VR1012 plasmid, designated as VR1C. Then the recombinant VR1C was entrapped with Chitosan nanoparticles prepared by the method of ionic cross linkage, and employed to inject muscularly 3-weeks old Kunming mice; the blank VR1012 packed with CNP and saline were used to inoculate mice as the control groups. 28 days after inoculation, all mice were orally fed with 0.4ml 2x108CFU/ per mouse virulent hemorrhagic enteritis E. coli to challenge the resistance against infection.

为研制安全高效免疫调节剂增强动物免疫抗病能力,本实验设计合成含11个C pG基序的寡聚核苷酸,重组构建含CpG的VR1012质粒(VR1C);制备壳聚糖纳米颗粒包裹重组质粒(CNP-VR1C),肌注接种3周龄昆明小白鼠,设壳聚糖包裹空质粒和生理盐水对照组;接种后28天口服大肠杆菌攻毒观察小鼠天然免疫的变化和对强毒感染的抵抗力,Sandwich ELISA测定血清免疫球蛋白和白细胞介素含量。

The results of MTT method In vitro culture renal tubular cells showed that the proliferous activity changed from sightly to dramatically intensed in response to fluoride concentration from 0.1mg/L to 15mg/L, but renal tubular cells proliferation obviously decreased in the 25mg F-/L group.

利用生物化学方法检测氧化应激指标,结果表明在钙营养充分的条件下,投氟100mg/L 16周大鼠肾组织GPX活性不是下降而是明显提高,脂质过氧化物变化不明显;低钙饲养条件下投氟100mg/L 16周大鼠抗氧化酶SOD活性亦有明显升高,而GPX却呈降低趋势。

At G. 32W stable expression of nov mRNA was distributed in pontine abducens nucleus, red nucleus and substantia nigra of midbrain, ventral posterolateral thalamic nucleus and mediodorsal thalamic nucleus.

第32周脑桥展神经核、中脑红核和黑质、丘脑腹外侧核和背内侧核以及第38周大脑纹状体、顶叶皮质显示nov mRNA稳定的表达。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank,选择包含人主要T细胞抗原表位序列的人PSCA基因片段,应用异种化抗原设计技术,保留人T细胞抗原表位,设计异种化PSCA融合抗原片段;(2)根据核酸序列按中心模板法设计引物,应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因,以PCR、限制性酶切和DNA序列测定法进行鉴定:(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点,采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4),并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中,转染293T细胞,借助免疫荧光+流式细胞术考察插入片段表达效率,最终选定PSCA_3片段进行下一步研究;(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下,插入pVAX1-neo-IRES—GM/B7载体中,构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB,并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况;(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞,制备小鼠前列腺癌模型,并采用股四头肌肌肉注射+电脉冲法(Electroporation,EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB,同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照,通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率,来评价该DNA疫苗在小鼠体内的抑瘤效果。

X-ray examination demonstrated that at 4 weeks after implantation, some or even lateral intertransverse bridging regions were unclear, and material density in the repairing group was lower than control group; at 8 weeks after implantation, interspace between up and down bridging was shrunk, and a great quantity of calluses were successively formed; at 12 weeks after implantation, complete confluence was observed, and the density in the repairing group was closed to that in the control group.

结果:采用脱蛋白处理后的松质骨可见大小不等、相互交通、开放孔隙的网架结构,孔径200~500 μm,孔隙率60%左右,其无机成分为羟基磷灰石,有机成分为胶原,力学性能保存良好,细胞相容性良好。X射线片结果显示:植入第4周,横突桥接处部分区域模糊,以内侧明显,此时修复组材料密度略低于对照组;第8周上下桥接部间隙变小,大量连续骨痂生成;12周后完全融合,两组材料密度接近。

Kanic acid was injected into guinea pigs' cochleae and the excitotoxicity model was established. After a week the recombinant plasmid was transferred into SGCs of guinea pigs' cochlea treated with HAT. The following week the expression of NT-3 was examined by the immunohistochemical method, and the morphology of SGNs was observed under the electronic microscope after 4 weeks, in the mean time the changes of auditory brain-stein response were examined.

通过耳蜗灌注海人酸(kainic acid, KA)建立豚鼠耳蜗兴奋性损伤模型,在给KA 1周后利用HAT携带重组质粒进行耳蜗灌注以转染耳蜗螺旋神经节细胞,免疫组化法观察转染后1周NT-3的表达及4周后电镜下螺旋神经节细胞形态学变化,同时观察对听觉脑干诱发电位(auditory brain-stem response, ABR)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。

Four weeks later, in the baicalin group, some spindle-shaped UCB-MSCs began to present shrinkage, with slender processes on the cell edge, and some UCB-MSCs tended to be spherical-, conical-, and triangle-shaped appearance, with many slender processes on the pseudopodia.

细胞形态学观察:脐血单个核细胞培养第2~3天开始贴壁生长,2周左右达高峰,3周后细胞生长达80%~90%融合,此时脐血原始间充质干细胞呈较均一的长梭形。4周后,黄芩苷组原来呈梭形的胞体一部分发生收缩,细胞边缘出现细的突起;一部分胞体逐渐近似圆形、锥形、三角形,伪足有多个细长突起。

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