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周质

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Results The hind limb function of the injured rats recovered at different degrees, the most extent recovery occurred during the time site 1~2 week, recovery continued from 2 to 3 week and, BBB was up to 12 at the end of the third week, but there was no significance recovery during 3~4 week. the astrocyte caudal to the injury plane began hyperplasy and hypertrophy; astrocyte in which GFAP in expression was positive the gray matter increased obviously from 3 days to 14 days after hemisection. The expression of MBP is same as that of GFAP.

结果 伤后后肢均有不同程度的恢复,1~2周时恢复幅度最大,2~3周时后肢运动功能继续恢复,3周时BBB评分最高达12分,3~4周运动功能无显著性恢复,损伤后1 d损伤远端3~6 mm处GFAP阳性星形胶质细胞开始增生肥大,3~4 d灰质中星形胶质细胞明显增多,2周时达到高峰,损伤近端3~6 mm处少突胶质细胞的增生反应过程与星形胶质细胞相似。

Recombinant plasmid pSVH 7 DNA of avian influenza virus H7 subtype heamagglutinin gene was encapsulated with DC-chol/DOPE liposomes and PC/chol/SA liposomes separately. Two-week old SPF chickens were intramuscularly inoculated with 50 μ g/0.2ml of the liposome entrapped PSVH 7 DNA. Four-weeks later, each chicken was challenged with 0.1ml 〓 AIV . One week after the challenge, the secretion of the cloacas was collected and transfected to chicken embryos to isolate the virus. The virus was isolated from 6/6 of the control group, 1/6 of the naked DNA group, 1/6 of the PC/chol/SA entrapped DNA group and 0/6 of the DC-chol/DOPE liposome entrapped group. The HI antibody titers (log2) of the four groups were 6. 83±0.98, 7. 0±1. 26, 7. 83±1. 17 and 8. 00±0.89 respectively 1-week after challenge, and 8. 5±0.55, 8. 17±0.82, 8. 68±0.45 and 9. 33±0.54 respectively 2-week after challenge. The results showed that inoculation of liposome entrapped DNA significantly enhanced resistance to virosis in animals.

将含禽流感病毒H7亚型血凝素基因的重组质粒pSVH7用DC-chol阳离子脂质体和胆固醇/卵磷脂/十八胺脂质体包裹,免疫2周龄SPF鸡,4周后用同型禽流感病毒进行人工感染,1周后采集泄殖腔分泌物分离病毒,结果未免疫组6/6分离到病毒,裸质粒DNA免疫组1/6分离到病毒胆固醇/卵磷脂/十八胺脂质体包裹DNA免疫组1/6分离到病毒,DC-chol脂质体DNA免疫组没有分离到病毒(0/6):人工感染后1周各组的HI抗体(Log2)分别为6.38±0.98,7.00±1.26,7.83±1.17,8.00±0.89,2周后为8.50±0.55,8.67±0.82,8.68±0.45,9.33±0.52,脂质体包裹组在同期均高于未免疫组和裸DNA免疫组,表明脂质体包裹质粒DNA免疫动物后,能增加动物对病毒感染的抵抗力和反应能力。

HRP was coupled with the fluorochrome FITC and encapsulated with liposome . pEGFP-N1 plasmid (expressing green fluorescent protein) with encapsulation in commercially available liposome was prepared. The eyes were enucleated 1, 4 and 8 weeks after zonule rupture and anterior segments comprising lenses were incubated in medium containing one of these components. Cryo-sections were made and translocation of fluorescent macromolecule from the medium into the lens and green fluorescent protein expression in the epithelium were observed by fluorescent microscopy.

制备FITC标记的HRP脂质体;制备pEGFP-N1质粒(表达绿色荧光蛋白的质粒),并用脂质体包裹;豚鼠悬韧带部分离断后1周、4周、8周取含晶状体的眼前段标本分别在含有FITC-HRP复合物、pEGFP-N1质粒的培养基中孵育,冰冻切片,荧光显微镜观察FITC-HRP复合物进入晶状体和pEGFP-N1质粒在晶状体内表达的情况,比较悬韧带离断侧与非离断侧的差异。3。

RESULTS AND CONCLUSION: PCHC: Intra-bone mechanical strength was lower at 2 weeks, the lowest at 4 weeks, but then closed to intension of normal cancellated bone at 8 weeks, higher than normal cancellated bone at 12 weeks, and recovered to the level of normal cancellated bone at 16 weeks.

结果与结论:多孔碳酸化羟基磷灰石骨水泥:2周时的骨内力学强度较低,4周时降到最低,8周时接近正常松质骨强度,12周时超过正常松质骨强度,16周时恢复到正常松质骨水平。

Coil and B. subtilis using the β-lactamase distribution assay. The results showed that the β-lactamase activities of E.coli are mainly in periplasm while the enzyme produced by B.

内酰胺酶活力测定表明,大肠杆菌产生的酶主要积累在周质空间,而枯草芽孢杆菌产生的酶主要分泌在胞外。

Western blotting confirmed that the low activity of b-lactamase in E. coli Top10 pcs+ resulted from a lower amount of b-lactamase in its periplasm.

由此可见,周质内低含量的b-内酰胺酶是导致E.coli Top10 pcs+细菌氨苄青霉素抗性降低的原因。

Expression level of rhGH could reach 250 mg/L culture solution by means of selecting components of original culture media, components of supplemental media, volume of supplemental media, the time of supplying supplemental media and supplying speed of supplemental media, secretion rhGH was found to represent 44%-54% of the total proteins in periplasm, and the fermentation time was reduced to 8 hrs.

通过对发酵培养基成分、补料成分、补料体积、补料时机和滴加流速的选择,分泌表达重组人生长激素的量可达到250mg/L,占周质蛋白的44%~54%,发酵周期缩短为8小时。

It shows that coexpression of secE/Y gene can increase a new band of about 25kD in the periplasm, but further electrophoresis analysis and Western blot prove that this band may not be the assembled TGF β1 dimer.

发现共表达secE/Y基因可使周质中新增25KD的蛋白带。

These results implied that processes catalyzed by these proteins probably occurred outside of cells, at least partially.

这些蛋白所参与的生化过程可能发生在膜外的周质空间或是菌体外的场所,为该物种营养的摄取,以及对环境做出响应服务。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length P1, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length P1, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3. 1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus , which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3. 1/IFN which includes the gene IFN-α of cattle. Subsequently, Recombinant plasmids were injected to cattles with or without pcDNA3. 1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies titers were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus,FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫苗,本研究通过定点突变方法和PCR扩增方法,获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D,将获得的基因片段直接/酶切后与同样处理的真核表达质粒连接,分别得到重组质粒pcDNA3.1/P12X3C和pcDNA3.1/P12X3C3D、pTARGET/P12X3C3D;对重组质粒进行序列测定、分析,并将重组质粒分别转染BHK-21细胞,通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达,用电子显微镜观察病毒空衣壳的组装;为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果,将重组质粒经肌肉注射方法接种豚鼠,并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1/IFN分别/同时免疫,第二次免疫后第三周豚鼠攻以1OOID〓或1000ID〓的O型FMDV China99株;随后将质粒pcDNA3.1/P12X3C、pcDNA3.1/P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1/IFN同时免疫牛,三周后经牛舌皮攻以10〓ID〓的O型FMDV China99株。

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