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多染细胞

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METHODS: Chemotactic chamber assay was used to investigate the regulatory role of formyl peptide receptor like-1 (FPRL1) in the chemotaxis of PMNs in response to LPS and PD.

用趋化小室方法测定虎杖甙对LPS刺激下多形核白细胞趋化活性及转染了甲酰化肽样受体(FPRL1)基因的293细胞(FPRL1/293)趋化活性的影响。

Alu dna was appeared on low level cam and no alu was found in control group.conclusion the results suggested that ph|20 may increase the palingenesis of blood vessel and stimulate human breast cancer cell growth and metastasis.

目前关于这方面的研究不多,本课题组曾将人ph|20 cdna转染到人乳腺癌细胞株mda231中,初步发现了ph|20可能通过肿瘤细胞释放fgf和促进血管生长来促进mda231增殖[5]。

However, the neoplastic cells show pleomorphism, with hyperchromatic nuclei.

肿瘤细胞的多形性明显,细胞核深染。

Microscopically, there are spindle to oblong cells with hyperchromatism and pleomorphism.

显微镜下见深染多形性的梭形或椭圆形细胞。

The neoplastic epithelial cells within the duct show minimal hyperchromatism and pleomorphism, but they have holes with sharp margins as though punched out by a cookie cutter.

导管内的肿瘤上皮细胞表现为淡染和多型现象,但是他们有边缘锐利的孔像被切饼机切过一样。

Results:17 rats were confirmed successful transplantation by MRI,the capacity of the tumor was 88.40±7.62mm3 on 8th day, 986.80±114.46mm3 on 16th day,the volume growth was 8-12 folds;on 8th day following transplantation,the tumor tissue was gray in color with fish homogen state without interior necrosis, the tumor cells were nest-distributed by HE staining,large dense staining of nucleus with manifest heteromorphism,tumor microvessel count increased by immunohistochemistry with buffy yellow staining, VEGF high expressed in the tumor cells with buffy yellow granule shape.

结果:17只大鼠经MRI检查证实均种植成功,肿瘤体积第8天为88.40±7.62mm3,第16天为986.80±114.46mm3,体积增长8-12倍之间;种植第8天,肿瘤组织呈灰白色、鱼肉均质状,内部未见坏死,HE染色肿瘤细胞呈巢状分布,胞核大浓染,异型性明显;免疫组织化学示肿瘤微血管数目较多,呈棕黄色染色,肿瘤细胞VEGF高表达,呈棕黄色细颗粒状。

Primary cultured chondrocytes are poly-angle, cytoplasm-rich, and their nuclei are either round or oval with clear necleole. Metachromatic and alcian blue positive staining in primary cultured chondrocytes was observed. Intercellular matrix was anti-collagen type Ⅱ staining but not anti-collagen type Ⅰ staining by IHC assay.

原代培养软骨细胞呈多角型,胞质丰富,胞核成圆形或椭圆型,核仁清楚,甲苯胺蓝呈异染性,阿尔新蓝8Gx 染色阳性,细胞外基质Ⅱ型胶原免疫组化染色阳性,Ⅰ型胶原染色阴性。

Chicken IL-15(ChIL-15), Which was discovered in 2001, plays a main role in activating NK cells and CD8+ memory T cells, and may therefore have potential as a vaccine adjuvant.According to published ChIL-15 gene sequence, a pair of primers were designed and synthesized to clone ChIL-15 cDNA from ConA-activated chicken splenocytes by RT-PCR. Recombinant plasmid pUC19ChIL-15 carrying the ChIL-15 gene was constructed. The gene encoding mature ChIL-15 (mChIL-15) was amplified from pUC19ChIL-15 by PCR and cloned into pMD 18-T vector. After digested with Kpn I /Pst I , the mChIL-15 gene was subcloned into prokaryotic expression vector pPRoEX橦Ta and transformed into E.coli DH5a .The transformant was induced by IPTG, and the recombinant ChIL-15(rChIL-15) was expressed as a fusion protein with a histidine hexamer tag at the N-terminal end of the protein. Following expression, the protein was purified by ProBond?Nickel-Chelating Resin. The uninduced and induced protein lysates and the purified protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane to identify rChIL-15 by Western blot. The rChIL-15 was used to immune the guinea pig to obtain rChIL-15 polyclonal antibody.

参考已发表的ChIL-15基因序列,设计合成引物,应用RT-PCR技术从刀豆球蛋白A活化的白来航鸡脾淋巴细胞中克隆ChIL-15 cDNA,将其与SmalⅠ酶切处理的pUC19载体连接,构建重组质粒pUC19ChIL-15;用PCR技术从pUC19ChIL-15中扩增出去信号肽的成熟ChIL-15(mChIL-15)基因,与pMD 18-T载体相连构建重组质粒pMDChIL-15;然后用KpnⅠ/PstⅠ双酶切下mChIL-15基因片段,定向亚克隆到经同样酶切处理的带有6个组氨酸标签的表达载体pP_RoEX~HTa中,构建重组质粒pP_RoChIL-15,对其测序确认读框正确后,转入大肠杆菌DH5α中诱导表达并进行纯化,用重组ChIL-15(rChIL-15)免疫豚鼠,制备多克隆抗体;再用HindⅢ/PstⅠ从质粒pP_RoChIL-15上切下mChIL-15基因,定向亚克隆到经同样酶切处理的杆状病毒转移载体pMelBacB中,经酶切、PCR和序列测定鉴定后,与杆状病毒线形化DNABac-N-Blue~(TM DNA共转染Sf9昆虫细胞,经三轮蚀斑纯化,构建重组杆状病毒rBac-ChIL-15,用该重组病毒感染处于对数生长期的Sf9细胞,按不同的时间收取样品,离心后对其上清和沉淀进行SDS-PAGE和Western blot分析。

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。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length PI, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length PI, 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-ELlSA 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 P 12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3.1/lFN which includes the gene IFN-a of cattle. Subsequently, Recombinant plasmids were injected to catties 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 liters 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疫曲,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100ID_(50)或1000ID_(50)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10~4ID_(50)的O型FMDV China99株。

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我的第一篇文章为屏幕力量杂志,在对这天是Jackie Chan 正式杂志,是香港actioner Gen X 警察的DVD 回顾,担任主角Nicholas Tse 和丹尼尔吴。

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