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The reaction between antigen and antibody was carried out by one step balance method and incubated in 4℃ for 24 hours, then separated bond and free antigen by PR reagent.

用氯氨T法制备125I标记IL-6,经Sephadex G-25纯化,抗原抗体反应采用平衡一步法,4℃温育24 h后经PR试剂分离结合和游离的标记抗原。

The reaction betwe en antigen with antibody was carried out by one step balance method and cultured i n 4℃ for 24 h,then separated binding and free antigen by PR reagent.

用氯氨T法制备125I标记IL-8,经Sephadex G-25 纯化,抗原抗体反应采用平衡一步法,4℃培养24 h后经PR试剂分离结合和游离的标记抗原。

We biopsied 1-2 single blastomere from 6-8 cell cleavage stage intracytoplasmic sperm injection embryo, and using nested PCR amplified the high frequently mutation region G380R of FGFR3 gene of the single blastomere. The products of PCR were digested by restriction enzyme Bfm Ⅰ, then the digested products were detected by 10% PAGE to see whether the embryo inherted the mutation of the patient and to screen out normal embryo transfer.

活检经胞质内单精子注射(intracytoplasmic sperm injection,ICSI)授精的胚胎发育至6~8细胞期的1~2个单卵裂球,采用巢式PCR扩增单卵裂球的FGFR3基因的高发突变位点G380R区域,用限制性内切酶BfmⅠ消化巢式PCR的内扩增产物,再经10%聚丙烯酰胺凝胶电泳检测有无遗传患者的该种突变,从而筛选出正常胚胎移植。

Swine enteropathogenic E. colis were rejuvenescenced and 6 of them were selected to prepare blood serum. We had tested the reactance of cross agglutination and agar diffusion between the strains and the blood serums. No. 300 and No. 343 were selected because they had highly cross reaction. To test superantigen of No.300 and No.343, the antigens made by both of them immunized piglets, and then we detected the serum 0D values with indirect ELISA. The OMP and PF of No. 300 and No. 343 were extracted for SDS—PAGE and westen-blotting.

本研究针对10株猪致病性大肠杆菌,经复壮后选取致病性强的6菌株作为供试菌株,制备免疫血清与这6菌株的多种抗原分别进行交叉凝集试验和琼脂扩散试验,根据各菌株抗原的交叉反应程度,筛选出抗原交叉程度高的菌株300和343,检测两菌株超抗原的毒性作用,以不同方式制备抗原免疫仔猪,用间接ELISA检测免疫效果,并提取菌株300和343的外膜蛋白和渗透因子经SDS—PAGE及免疫印迹试验,进行相关分析。

Results: 29 of 32 residual ulcer wounds had healed in 3 weeks. Rest 3 cases were continually treated with MEBO and dipping bath to control the infection so as to make the size of the wounds deceased. In case of the area of a wound X4 cm, i. e. the area was more than 5% TBSA, the wound was skin-grafted in a way of stamp and then was applied with MEBO continually till healing.

结果:32例患者中,29例残余创面均在3周内愈合,另3例经应用MEBO及浸浴治疗感染基本控制后,创面缩小,单个面积仍在4cm×4cm以上者,每例残余创面面积均在5%TBSA以上,经邮票植皮后继续用MEBO治愈。

Methods 14 mutanted vpr fragments selected from Chines patients with HIV. Both eukaryotic expression vector pcDNA3.1 and PCR products purified , double-cut by HindⅢ and BamH and the cut products legated and were into competent cells JM109. The 14 reconstructed plasmids electronically transfected into hela cells, and established cells with pcDNA vprwt 、pcDNA vpr-Fs and pcDNA3.1 blank cells, and without pcDNA3.1 cell.

将14个带有HIV-1 vpr基因片段的pcDNA3.1真核表达载体构建重组质粒,将其转染Hela细胞,并设立保守株vp r基因转染细胞、突变株 vpr-FS基因转染细胞、空载体转染细胞和未转染细胞作为对照,经RT-PCR检测目的基因转染成功后,经Pi染色用流式细胞仪检测被转染细胞的细胞周期分布和细胞凋亡率。

After a muliobjective searching by AT-0520 D electronic computer using orthogonal regression design, the optimum technological conditions including the principal factors which influence the chemical reactions have been found. The expected yield of butyl acrylate, the content of byproduct β ester and acid value are all identical with the experimental data.

对影响反应的主要因素,采用正交回归设计和用AT-0520D电子计算机对多目标进行搜索,得出优化工艺条件组合,经实验验证,丁酯收率、付产β酯含量、酸值的予测值都和实验值吻合,并经生产规模验证,得到满意结果。

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。

Using 2MHz pulseultrasonic Doppler probe, the index of hemodynamics such as blood flow rate、audio frequency、vascular pulsation of bilateral vertebral and basilar arteries were detected in order to analyze the diagnosis of cervical spondylosis involoving vertebral artery by Transcranial Color Doppler through the occipital foramen. Results: of all 108 carses, 80 cases were positive(positive rate 74.7%). In CSA group,cases of low flow rate were 45.6%,cases of high flow rate were 28.7%,and cases of no...

为分析经颅多普勒检测椎动脉型颈椎病的诊断价值,采用配对研究方法,用 2MHz脉冲多普勒探头经枕骨大孔窗口,检测左、右椎动脉及基底动脉的血流速度、流向、音频、血管博动指数等血流动力学指标,结果显示 10 8例患者有 80例异常,异常率 74 7%,并发现CSA组低流速者占 45 4 %,高流速者占 2 8 7%,正常流速者占 2 5 9%,与正常对照组间有显著差异(P 0 1),总体而言,低流速型是CSA的主要特征之一。

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