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The ELISA assay established with the crude antigen-specfic monoclonal antibodies could detect both of the clinical and environmental isolates of Aspergllius, while the other assay could only detect Aspergillus fumigatus of both clinical and environmental isolates.And no cross reaction with the cell culture of Penialllium marneffei and Candidas was observed with the two methods.

用mAbs-1建立的双抗体夹心ELISA法可检测19种常见曲霉株培养液;用特异性针对烟曲霉抗原单克降抗体(mAbs-2)建立的双抗体夹心ELISA法可特异性检测临床和环境分离株烟曲霉培养液;与其他曲霉株无交叉反应;2种双抗体夹心ELISA法与马尔尼菲氏青霉菌及念珠菌培养液均无交叉反应。

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。

The plasma and serum DNA with differnet dilutions were quantified to evaluate the stability of this assay. The repeatability of this assay was also evaluated.

针对不同程度稀释的血浆血清DNA样品进行定量检测,确定检测方法的稳定性,并对该检测方法的重复性进行评价。

There was no satistically significant difference among the quantitative results of plasma and serum DNA samples with diffemet dilutions (P<0.05). The intra-assay and extra-assay CV were 11% and 17%, respectively.

对于不同稀释程度的血浆血清DNA样品(分别稀释为原浓度的50%、10%和1%),定量检测结果与未稀释DNA样品的差异无统计学意义(P<0.05)。

JP2Rhodium of 10-9 level was accurately measured by NAA combined with lead fire assay, and 10-10 level was accurately measured by NAA combined with nickel sulfide fire assay.

铅试金中子活化法可实现10-9铑含量的准确测量,镍锍试金中子活化法可实现10-10铑含量的准确测量。

In BP exposure experiment, except that the neutral red assay showed that BP significantly inhibited the haemocyte viability, the other methods did not detect the inhibition effect, besides, the results of the neutral red assay showed that the inhibition degree had a positive correlation with the concentration and exposure time of BP.

采用中性红比色法检测BP对栉孔扇贝血细胞活性具有显著抑制作用,且抑制程度与BP作用浓度、时间呈正相关,其余方法均未检测出BP对血细胞活性的影响。

DNA synthesis rate was assessed by performing tritiated thymidine incorperation assay. The expression of SM-α-actin and proliferation of DSMC were analyzed by immunofluorescent assay and flow cytometory.

结果显示,逼尿肌细胞的形变量和粘弹性并不呈线性关系,一定限度内的张力负荷(拉伸20%)粘弹性系数下降,而继续加大张力(拉伸30%)时变化并不明显。

Methods: rabbit bone marrow mesenchymal stem cells were purified, and then cultured in inducing medium containing transforming growth factor-β1; mtt assay was employed to evaluate the proliferative activity of induced cells; immunohistochemical assay was used to detect the expression of type ⅱ collagen; induced cells were transplanted into the knee of rabbit, and x-ray photography was employed to observe the repair and formation of cartilage.

纯化兔骨髓间充质干细胞;用含转化生长因子β1的诱导液培养骨髓间充质干细胞;以mtt测定诱导细胞的增殖活性;免疫组化检测ⅱ型胶原蛋白的表达;诱导后的细胞移植于白兔膝关节内,x线摄片观察软骨修复形成情况。

The proliferation of Schwann cells of sciatic nerves was determined in different time by MTT assay and Thymidine incorporation assay.

结果 人参皂甙 Rb1 在 10μg/ ml的浓度对雪旺细胞增殖有明显促进作用,高浓度的人参皂甙 Rb1 1mg/ ml则显示抑制作用。

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The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

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