特异性

The primary conclusions of the present work are as following first, the charges of membrane sarface effect the rate of protein adsorption, but do not effect the amount of protein ultimately adsorbed second, the nonspecific adsorption of avidin can be detached by adding a suitable amount of bivalent alkaline-earth metal ions; third, the binding of avidin to membrane bound model receptor is astricted remarkable by the latelal steric hindrence.

实验结果表明，膜表面电荷对非特异性吸附的速率有很大的影响，而对最终的蛋白吸附量影响不大；非特异性的吸附可以通过二阶阳离子的脱附而去除；受体蛋白质与配体间的特异性结合受到侧向空间位阻效应的制约。

These specific cDNAs and DNAwere subcloned into vector for sequencing.The sequencing results demonstrated thatthe specific cDNA from P.chinensis eyestalk consists of 203 base pairs,and all thespecific cDNAs from the shrimps T.curvirostris,P.stylirostris and the crab E.sinensis consist of 215 base pairs.

分别将这些片段亚克隆到载体中进行测序，测得中国对虾特异性cDNA片段由203个碱基组成，中华绒螯蟹、鹰爪虾、蓝对虾的特异性cDNA片段及蓝对虾的特异性DNA片段由215个碱基组成，其中蓝对虾的特异性cDNA与由蓝对虾的基因组DNA扩增得到的特异性DNA片段的碱基序列几乎完全相同。

The protocol for the detections of all the AIV isolates, the H5-subtype and the optimization of reactions were studied with a H5-subtype reference isolate BSG1. A 229bp-band of AIV specific and a 380bp-band of H5 subtype specific were amplified individually with the designed primers of AIV specific and H5-subtype specific respectively by the developed RT-PCR technique from isolate BSG1. A double-PCR was also developed which can detect all the AIVs and identify the H5-subtype AIVs. The results of specificity experiment showed that there was no specific band could be amplified with the samples of Newcastle disease virus, infectious bronchitis virus, infectious laryngotracheitis virus, egg drop syndrome virus and infectious bursal disease virus.

结果应用通用引物可从BSG1毒株扩增到与预计大小相符的229bp AIV特异性片段，应用H5亚型鉴定引物则可从BSG1毒株扩增到与预计大小相符的380bp H5亚型特异性片段；在此基础上建立了可检测所有AIV毒株以及可同时进行H5亚型毒株鉴定的二重RT-PCR方法；检测方法的特异性试验结果表明，该方法对新城疫病毒、传染性支气管炎病毒、传染性喉气管炎病毒、产蛋下降综合症、传染性法氏囊病毒无交叉反应；检测方法的敏感性试验结果表明，通用引物扩增、H5亚型特异性引物扩增以及二重PCR扩增反应的组织样品总RNA的最低检出量分别为0.0217μg、0.0217μg和2.17μg；对20份临床样品检测中，有10份为AIV阳性，其中有5份为H5亚型病毒阳性。

The larva-specific expression level of the function correlated to celluar components was the highest.The highest specific expression level of molecular function was appeared at neurula stage.A lot of functional specific expression about biological process was also appeared at neurula stage.7.The result showed that the pearson correlation coefficient was 0.896 compared between the sea urchin and Ciona savignyi.

进一步分析发现：在与细胞组件相关的功能特异性表达方面，幼体期的特异性表达量最高；在与分子功能活性相关的功能特异性表达中，神经胚时期的特异性表达量最高；而参与生物过程的功能特异性表达方面，神经胚时期的特异性表达量则最高。7不同物种间转录组分析显示尾索动物海鞘的转录组与棘皮动物海胆的最接近，其相关系数为0.896，而海胆与文昌鱼的相关系数只有0.682，结果表明在进化上文昌鱼可能与脊椎动物更接近。8运用Apache、php、MySQL技术搭建了Widows平台的文昌鱼EST分析数据库。

In this dissertation, firstly, the conditions of isolation and purification of specific toxin fractions produced by Exserohilum turcicum has been studied, and a high-efficient method has been established;secondly, the toxins were analysed by UV-Vis spectrophotometry;then the effect of specific toxin on death rate of corn leaves protoplast was studied by FDA as a tinct reagent;finally, using ANS as a probe and SDS-PAGE to study the membrane protein of the corn leaves with Htl gene. In a word, the aim of the research is to search some existent evidences of the toxin binding site on protoplasmic membrane, and provide proofs for nosogenesis of specific toxin in terms of cytology and the molecular biology.

本研究对玉米大斑病菌1号小种毒素的特异性组分进行分离，建立了高效的特异性毒素分离纯化方法，并对其进行了紫外波谱分析；用荧光素双醋酸酯染色法，测定了毒素特异性组分对原生质体死亡率的影响；进而用1-苯胺基-8-萘磺酸荧光探针标记法和SDS-PAGE电泳法对特异性毒素组分与Htl基因玉米细胞互作过程中的质膜蛋白进行了研究，旨在证明Htl基因玉米细胞质膜上存在特异性毒素组分结合蛋白，为从细胞和分子生物学角度阐明毒素的致病机理提供证据。

DCs acquired by our reformed methods express both CD83 and CD 14 molecules highly, and have a higher density than other domestic reports. The higher TNF in DCs culture medium of HC patient suggests DCs in patient still have antigen presenting ability and by optimization the culture medium would improve its presenting ability and have a potential value in design and application individual vaccine. Although antigens pulsed DCs have a decrescent antigen presenting ability but BCG HSP70 could induce its mature and improve its presenting ability. Suggests BCG HSP70 would be a useful mature inducer. Lymphocytes primed by DCs based HC vaccine have the specific cytotoxicity against HCC lines. The CTL after freezing and anabiotic could prophylaxis and therapy HC xenograft on nude mouse. The results also suggests that CD4〓 lymphocytes play a important role in HC with a good differentiation and would be useful in treatment this kind of HC. After being activated by Peptide LLNQHACAV of hAFP and apoptotic HCCs pulsed DCs respectively, the culture medium of activated lymphocytes both contains a high level Th1 cytokines IL-12 and TNF. Primed lymphocytes appeared a characteristic of NK cells. DCs not only inhibited the growth of human HCC and other cancer cells in vitro but also prevented the growth of HC xenograft on nude mouse in vivo. There are at least three kinds of mechanism playing important role in DC based vaccine ,there are inhibition of DCs, HC specific CTL and cytokines pathway.

诱导出的DC共同表达CD83和CD14分子，CD83分子表达明显高于国内报道；肝癌患者DC培养上清中TNF水平高于健康人，提示肝癌患者DC仍具一定的抗原呈递能力，适当调控可使其行使APC功能，以期在肝癌个体化疫苗中发挥作用；DC负载肝癌可溶性抗原后，抗原呈递能力降低，BCG HSP70可促进DC成熟，增加其抗原呈递能力，预示BCG HSP70有可能成为促进DC活化和成熟的另一重要分子；肝癌DC疫苗在体外诱导肝癌特异性淋巴细胞，活化的淋巴细胞在体外对肝癌细胞的杀伤以特异性CTL为主，同时分泌较高水平Th1型细胞因子IL-12和TNF，并抑制4种肝癌细胞生长；冷冻复苏后的肝癌特异性淋巴细胞可以预防和抑制人肝癌裸鼠皮下移植瘤，提示DC负载肝癌可溶性抗原后诱发的MHC-Ⅱ类限制性CD4〓T细胞有可能在分化程度高的肝癌治疗中发挥作用；用DC和HLA-A2〓DC分别负载凋亡肝癌细胞和hAFP218-226位LLNQHACAV HLA-A2限制性九肽，在体外诱导肝癌特异性淋巴细胞，活化后的CTL细胞分泌较高水平的Th1型细胞因子IL-12和TNF，并具较强杀伤活性，此CTL同时具备NK细胞特征；DC对肿瘤细胞的抑制作用可能是通过吞噬实现的，Fas-L在DC抑制中也起一定作用；DC对人肝癌裸鼠皮下移植瘤的抑制率为97％；在肝癌DC疫苗的作用中，至少联合3种以上机制，即通过DC的直接作用、肝癌特异性CTL和细胞因子途径直接或间接地杀伤和抑制肝癌细胞。

Get high purity DCs by Cultured plastic-adherent monocytes isolated from healthy human peripheral blood with GM-CSF and IL-4 for 7 days. To observe the morphology of DCs by inverted phase contrast microscope ,electron microscope and laser confocal microscope. Analyse phenotype of DCs with flow cytometry. Investigate the endocytosis ability of DCs as a group by Horseradish peroxidase endocytosis assay. To appraise allogeneic mixed lymphocytes reaction of DCs by MTT reduction assay. Analyse the levels of IL-12 and TNF in liquids of cultured medium by ELISA and MTT reduction assay respectively. Soluble antigens of HCCs was obtained by 3 freeze-and-thaw cycles. Biological characteristics of HC soluble antigens pulsed DCs were monitored by flow cytometry. According to MTT reduction assay estimated the cell proliferation of self lymphocytes activated by HC antigens pulsed DCs. Get high purity BCG HSP 70 protein by SDS-PAGE electrophoresis and determined its biological activity with ELISA. Analyse phenotype of antigen pulsed DCs primed by BCG HSP70 with flow cytometry. By MTT reduction assay estimated the cell proliferation of self lymphocytes and the MLR of DC based vaccine. Analyse expression of HLA-DR molecule on surface of HCC lines. The IFN-γ mRNA in lymphocytes after actived by DC vaccine and the Fas-L expression on DC and DC vaccine primed lymphocytes were detected by in situ hybridization and flow cytometry respectively. Specific cytotoxity lysis of T lymphocytes and nonspecific inhibition of liquids in culture medium against HCC lines were also tested. Detect expression of hAFP on four HCC lines with Cell-ELISA. Induce apoptosis of HCCs with actinomycin-D. Interaction of DCs and apoptotic cells was observed under transmission electron microscope. Growth inhibition test of DC against HCC lines was also performed. Establish the nude mouse model bearing human HC xenografts and indentify the characteristic of tumour by histochemistry and immunohistochemistry techniques. Prevent and treat transplanted human HC on nude mouse with Freezing and anabiotic HC specific lymphocytes.

用GM-CSF和IL-4从健康人外周血诱导DC；分别用倒置相差显微镜、电子显微镜及激光共聚焦显微镜观察DC形态；流式细胞术检测DC表型；HRP吞噬实验测定DC的群体内吞能力；MTT法检测同种异体混合淋巴细胞反应；ELISA法和MTT法分别测定DC培养上清液中IL-12和TNF水平；冻融法制备肝癌细胞可溶性抗原；流式细胞术检测负载肝癌可溶性抗原后DC的生物学特性；MTT法检测DC负载肝癌抗原后对自身淋巴细胞增殖的影响；SDS-PAGE制备电泳纯化BCG HSP70并鉴定纯度，ELISA测定活性；流式细胞术检测负载抗原DC经BCGHSP 70活化后的表型；MTT法检测肝癌DC疫苗对自身淋巴细胞增殖的影响和混合淋巴细胞反应；流式细胞术检测肝癌细胞表面HLA-DR表达；MTT法检测肝癌DC疫苗对自身淋巴细胞的活化；原位杂交法检测肝癌DC疫苗活化后的淋巴细胞IFN-γmRNA表达；流式细胞术检测DC和肝癌DC疫苗活化后淋巴细胞表面Fas-L；MTT法分别检测肝癌DC疫苗活化的淋巴细胞和其培养上清对肝癌细胞的特异性杀伤和非特异性抑制作用；Cell-ELISA检测人肝癌细胞hAFP表达；MTT法检测负载AFP表位肽和凋亡肝癌细胞DC对自身淋巴细胞增殖的影响；ELISA法和MTT法分别测定活化后淋巴细胞培养上清中TNF和IL-12水平；肝癌细胞凋亡的诱导和检测；DC吞噬凋亡肝癌细胞后的电子显微镜观察；DC对肝癌细胞的生长抑制试验；人肝癌裸鼠皮下移植瘤动物模型的建立及其组织学和免疫组织化学鉴定；DC及肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤；冻存和复苏后的肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤。

Results The sensitivity,specificity,positive predictive value,negative predictive value,Youden index,accuracy ...

结果检测血清特异性IgG的灵敏度为100·0%，特异度为97·7%，阳性预测值为97·2%，阴性预测值为100·0%，约登指数为0·977，符合率为98·7%，试验的一致率为98·5%；检测血清中总的特异性Ig灵敏度为97·1%，特异度为95·3%，阳性预测值为94·4%，阴性预测值为97·6%，约登指数为0·925，符合率为96·2%，试验的一致率为97·8%；检测唾液中特异性sIgA的灵敏度为96·2%，特异度为94·5%，阳性预测值为89·3%，阴性预测值为98·1%，约登指数为0·907，符合率为95·1%，试验的一致率为97·5%；检测粪便标本中的特异性sIgA的灵敏度为92·0%，特异度为90·2%，阳性预测值为85·2%，阴性预测值为94·9%，约登指数为0·822，符合率为90·9%，试验的一致率为98·6%，CV值均小于15%。

objective to establish immunological methods specific for detecting antigens in different groups of monoclonal antibodies.methods indirect immnofluorescence assay was applied to identify specificity of the two groups of monoclonal antibodies prepared with crude antigen and recombinant antigen of aspergillus fumigatus,respectively.two different double monoclonal antibody sandwich elisa assays established with the two groups of antibodies were performed to detect antigents in the cell culture supermatants of 19 common species of aspergillus,penicillium marneffei,and 5 species of candidas.results the results of indirect immnofluorescence assay indicated that the monoclonal antibodies prepared with crude antigen of aspergillus fumigatus were specific for antigens in both clinical isolates and environmental isolates of aspergillus, whereas the other group of monoclonal antibodies was proved to be specific for aspergillus fumigatus of both clinical and environmental isolates.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.conclusion the elisa assays can detect both of the clinical and environmental isolates of aspergillus,and differentiate aspergillus fumigatus from other species of aspergillus.

目的 用2组曲霉单克隆抗体建立特异性识别不同种类曲霉抗原的检测方法。方法采用天然烟曲霉抗原免疫，获得广谱针对曲霉抗原的单克隆抗体；采用重组烟曲霉抗原获得特异性针对烟曲霉抗原的单克隆抗体，用间接免疫荧光鉴定，并分别建立2种双抗体夹心elisa法，对19种常见的环境和临床分离曲霉株、马尔尼菲氏青霉菌及念珠菌培养液进行检测。结果间接免疫荧光显示，用天然烟曲霉抗原免疫获得的单克隆抗体(mabs-1)可广谱识别多种曲霉分离株，而重组烟曲霉抗原获得的单克降抗体(mabs-2)仅能特异性结合临床和环境分离的烟曲霉抗原。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。

The result showed HSV seropositive mice was really induced to produce tumor special or non-special factors, possibly included natural killer cell and lymphokine activated killer cell, which cooperated with mtHSV to kill tumor cells in course of mtHSV mediated tumor therapy. The effect in wtHSV seropositive mice was better than in mtHSV seropositive mice.

实验证明HSV免疫小鼠确实诱导产生了肿瘤特异性或非特异性因子，可能包括NK细胞和LAK细胞(lymphokine activated killer cell)，这些肿瘤特异性或非特异性因子在mtHSV介导的肿瘤治疗中与mtHSV一起协同杀死了肿瘤细胞，HSV-1F免疫小鼠的这种作用强于mtHSV免疫小鼠。

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务，乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐，因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。