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Methods: A model to evaluate lymphocyte proliferation stimulated with a polyclonal activator, concanavalin A, was established by vital dye carboxyl fluorescin diacetate succinmidyl ester labeling technique. Effects of the different doses of anisomycin on the lymphocyte proliferation were estimated by flow cytometry and MTT methods. The propidium iodide labeling technique was applied to assay the effect of the different doses of anisomycin on changes of the lymphocyte cell-cycle stimulated by ConA or by phorbol ester plus Ionomycin. The percentage of the expression level of CD69 and CD25 on the activated lymphocytes was evaluated by fluorescin-conjugated monoclonal antibody double labeling technique.

以活体染料羧基荧光素乙酰乙酸琥珀酰亚胺酯染色,建立在多克隆刺激剂刀豆蛋白A刺激下评价小鼠淋巴细胞增殖的模型,通过流式细胞术和MTT法分析茴香霉素在不同剂量下对淋巴细胞增殖的作用;采用碘化丙锭染色分析茴香霉素对ConA或佛波醇酯加离子霉素刺激的小鼠淋巴细胞周期变化的作用;利用荧光标记的单克隆抗体双染技术和流式细胞术观察茴香霉素对小鼠CD3~+T细胞早期及中期活化标志分子CD69和CD25表达的影响。

Abstract] Objective To Treating the nano-SiO2 particle by silicon coupling reagent, functionalize the SiO2 by making double bond on its surface. Polymerizing on the surface of nano-SiO2 surface by the method called emulsion polymerization, the nano-SiO2 is covered by polymer to get the PMMA-KH-570-nano-SiO2 complex material. Test the surface structure of the surface of nano-SiO2 by FTIR, and test sort of its mechanical performances.Comparing the treated nano-SiO2 which is directly dispended in the PMMA emulsion with the SiO2 without any treatment, we are trying to find an effective method which can not only increase the dispersion of SiO2, but also improve the mechanical performance of the artificial teeth.

目的 通过硅烷偶联剂KH-570处理SiO2纳米粒子,向其表面引入双键使其功能化,采用乳液聚合的方法在SiO2纳米粒子表面进行接枝聚合,实现高分子对SiO2纳米粒子的表面包覆处理,制得PMMA-KH-570-SiO2纳米复合材料,采用FTIR对SiO2纳米粒子表面结构进行了表征研究,并且测定了它的多种机械性能,与表面处理过的SiO2纳米粒子直接分散至PMMA乳液中制得的复合材料、未添加任何增强物质的空白组进行比较,探索一种能够提高SiO2在基体中的分散性,为义齿复合材料的研究和开发提供理论依据和参考。

Methods: animals were randomly classified into 5 groups: group a, gaster poured by physiological solution; group b, gaster poured by ethanol; group c, gaster poured by ethanol after hypophrenic vagectomy; group d, gaster poured by ethanol after big small splanchnicotomy; group e, gaster poured by ethanol after hypophrenic vagotomy and big small splanchnicotomy.the distribution of c fos positive neuron in central nervous system was displayed and immunohistochemical staining method was used.

取30只成年雄性wistar大鼠,随机分为5组,每组6只。a组:单纯生理盐水灌胃组;b组:单纯乙醇灌胃组;c组:膈下迷走神经切断乙醇灌胃组;d组:内脏大、小神经切断乙醇灌胃组;e组:膈下迷走神经和内脏大、小神经均切断乙醇灌胃组。运用免疫组化技术显示fos阳性神经元在大鼠中枢神经系统中的分布。

Methods: Animals were randomly classified into 5 groups: group A, gaster poured by physiological solution; group B, gaster poured by ethanol; group C, gaster poured by ethanol after hypophrenic vagectomy; group D, gaster poured by ethanol after big small splanchnicotomy; group E, gaster poured by ethanol after hypophrenic vagotomy and big small splanchnicotomy.

取30只成年雄性Wistar大鼠,随机分为5组,每组6只。A组:单纯生理盐水灌胃组;B组:单纯乙醇灌胃组;C组:膈下迷走神经切断乙醇灌胃组;D组:内脏大、小神经切断乙醇灌胃组;E组:膈下迷走神经和内脏大、小神经均切断乙醇灌胃组。

Drug sensitive test and three-dimensional test220 strains of Pa were isolated from hospitalized patients between 2003 and 2007. K-B method was used to tested the susceptibility of 10 different antibiotics. IRPa was screened by testing the minimal inhibitory concentration of imipemem by using agar diluiion method.The susceptibility of these IRPa to the antibiotics was analysised. Three-dimensional test was used to identify the different kinds of beta lactamases from 220 strains of Pa.2.Carbarpenems hydrolytic enzyme genes and oprD2 gene were detectedamong the selected IRPa strains, PCR method was performed to detect carbapenemase genes which included GES、KPC、SPM、VIM、IMP、GIM gene and the oprD2 gene;Multiplex PCR were used to detect OXA genes and plasmid-mediated AmpC beta lactamase genes; The expression of the chromosomal AmpC beta lactamases and oprD2 genes in IRPa strains were analyzed by Real-time PCR.3.Identification and characterization of integronsIntegrase gene was detected by PCR, and the classification of integrons was performed by using restriction fragment length polymorphism.PCR was performed to detect the qacE△1-sull gene,and the gene cassetes which are located at variable region of integrons in the strains were detected to be positive.

方法1、药敏实验和三维实验收集2003~2007年临床分离的220株Pa,对这些菌株采用K-B法测定10种临床常用抗生素的药敏情况,同时采用琼脂稀释法检测亚胺培南的最低抑菌浓度(Minimal inhibitory concentration,MIC),筛选出对亚胺培南耐药的铜绿假单胞菌,并分析其对其它抗生素的药物敏感率;采用三维实验的方法分析220株Pa产β内酰胺酶的类型。2、碳青霉烯类水解酶和oprD2蛋白的检测针对鉴定的IRPa菌株,采用普通PCR方法检测具有碳青霉烯水解作用的β内酰胺酶耐药基因(GES、KPC、SPM、VIM、IMP、GIM基因)和oprD2基因,采用多重PCR的方法检测OXA型基因和质粒携带的AmpC酶基因,用荧光定量RT-PCR方法检测oprD2蛋白基因表达情况;同时对产AmpC酶的Pa(25株,含IMP耐药和敏感株)用RT-PCR方法检测AmpC酶基因的表达量情况。3。

METHODS AK was purified by DEAE ion-exchange cellulose(DEAE-52) and sepharose Cl-4B .The component of the saccharide was determined by GC and TLC.Its MW was measure d by gel filtration.The form of glucosidic bond was detrermined by IR.The conten t of the saccharide was measured by colorimetry.

采用DEAE纤维素(DEAE-52)柱层析和凝胶柱层析分离纯化草乌多糖,气相色谱法和薄层层析法测定其多糖组成,凝胶过滤法测定其相对分子质量,红外光谱测定其苷键类型,比色法测定其糖含量。

In this study, different nano-hydroxyapatite particles,HAP_1(25-60nm), HAP_4(the additives is heparin, 15-50nm), HAP_5(the additives is bovine serum albumin BSA, 20-80nm) were prepared by homogeneous precipitation and used heavy-gauge hydroxyapatite as comparison,we determined amount of heparin, sialic acid ,BSA adsorbed on HAP_1,HAP_2,HAP_3 by Crystal Violet assay, Bialsche method,Bradford method respectively and analyzed the binding mechanism by infrared spectrum;After taking HAP_1,HAP_2,HAP_4,HAP_5 and RBC to co-culture in vitro,we studied RBC hemolysis test and detected RBC hematolysis rate by erythrocyte osmotic fragility test;observing the changes of morphous and locomotion of cell after coacting HAP_1,HAP_2,HAP_4,HAP_5 and RBC by light microscope and inverted phase contrast microscope;observing HAP_1,HAP_2,HAP_4,HAP_5 effecting on Ultrastructure of RBC.

本文用均匀沉淀法制备了HAP_1(25-60nm),HAP_4(添加剂肝素,15-50nm),HAP_5(添加剂牛血清白蛋白BSA,20-80nm)等纳米粒子,并用大尺寸的羟基磷灰石HAP_2(470-520nm),HAP_3(1906nm)作对照,分别利用结晶紫法、Bialsche法、Bradford法研究了肝素、唾液酸、血清白蛋白在HAP_1,HAP_2,HAP_3上的吸附量,用红外光谱分析其中的结合机理;在体外将HAP_1,HAP_2,HAP4_,HAP_5与红细胞共培养,进行了红细胞溶血试验的研究,并借助红细胞渗透脆性试验检测红细胞溶血率;运用普通光学显微镜和倒置相差显微镜观察了HAP_1,HAP_2,HAP_4,HAP_5与红细胞作用后细胞形态及运动的变化:透射电镜观察了HAP_1,HAP_2,HAP_4,HAP_5对红细胞超微结构的影响。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank,选择包含人主要T细胞抗原表位序列的人PSCA基因片段,应用异种化抗原设计技术,保留人T细胞抗原表位,设计异种化PSCA融合抗原片段;(2)根据核酸序列按中心模板法设计引物,应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因,以PCR、限制性酶切和DNA序列测定法进行鉴定:(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点,采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4),并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中,转染293T细胞,借助免疫荧光+流式细胞术考察插入片段表达效率,最终选定PSCA_3片段进行下一步研究;(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下,插入pVAX1-neo-IRES—GM/B7载体中,构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB,并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况;(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞,制备小鼠前列腺癌模型,并采用股四头肌肌肉注射+电脉冲法(Electroporation,EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB,同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照,通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率,来评价该DNA疫苗在小鼠体内的抑瘤效果。

Soil Ca2+ content increased by 0.749mmol/kg, Mg2+ content increased by 0.169mmol/kg, the Na++ K+ content dropped by 9.52mmol?kg, SO42- increased by 0.091mmol?kg, Cl- reduced by 1.727mmol?kg, the HCO3- content in the soil assumed the obvious dropping tendency, it might be explained that the advancement of HCO3- has transformed into CO32- in the oasis, and has formed precipitation with two price calcium magnesium heteropolar ,then fix in the soil layer.

4土壤Ca2+含量增加了0.749mmol/kg,Mg2+含量增加了0.169mmol/kg ,Na++K+含量下降了9.52 mmol∕kg,SO42-增加了0.091 mmol∕kg, Cl-降低了1.727 mmol∕kg,土壤中HCO3-的含量呈明显下降趋势,可解释为在绿洲化进程中HCO3-转化为CO32-,并与二价钙镁离子结合形成沉淀固定于土层中。

According to the variation in the components of intermediateacid igneous rocks coming from the lower crust, it is possible to restrain the depth of the source area and the minimum thickness of the crust, and thus to provide important information for the study of the deep process of the continental intraplate mineralizationGlobally, many world level porphyry copper deposits and metallogenic systems formed by hypabyssal hot liquid have their close relationship with the synchronous adakite in spacetime and genesis; domestically, the adakitelike rocks have been identified to be related to the metallogenesis in the main metallogenic areas in ChinaThe recognition of adakitelike rocks having no relationship with the process of subduction makes it possible to construct a metallogenic model of continental intraplate porphyry metal deposits by combining other geological evidences, and this model is totally different from the metallogenic model of porphyry copper deposits with Bsubduction setting constructed by Sillitoe (1972)The existence of adakitelike rocks may be the necessary condition but not the sufficient one for forming the largescale porphyry deposits and the hypabyssal hotliquid deposits, whose metallogenic elements mainly came from the mantleThe metallogenic potential of adakitelike rocks is achieved by the entering of the mantle material, and the metallogenic specialization of adakitelike rocks is decided by the distribution characteristics of the metallogenic elements in the upper mantleAn important reason for the adakitelike magma related to subduction being advantageous to mineralization is that there were abundant high pressure and high temperature liquid coming from the subducted platepieces and the magma of high fO2 coming from the melting of subducted platepiecesHowever, for those adakitelike rocks, whose occurrence has continental plate background but does not relate to the subduction, their metallogenic mechanism is not clear yetBased on the concept of modern mineral exploration and combined with the analysis of integrated geological information, we may better realize the practical prospecting significance of the adakitelike rocks

根据起源于下地壳的中酸性岩浆岩的成分变化,可以约束其源区深度以及地壳最小厚度,为大陆板内成矿作用的深部过程研究提供重要信息。全球范围内,许多世界级斑岩铜矿和浅成热液矿化系统与同期的埃达克质岩存在密切的时空与成因联系,在国内主要成矿区带也识别出与金属成矿作用有关的埃达克岩。与俯冲过程无关的埃达克质岩的识别,使我们有可能结合其他地质证据构建完全不同于Sillitoe(1972)B型俯冲环境的斑岩铜矿成矿模式的大陆板内斑岩型金属矿床成矿模式。对于规模巨大、矿质主要源自地幔的热液矿床的形成,埃达克岩可能是必要条件,但不是充分条件。埃达克岩的成矿潜力通过地幔物质加入而获取,埃达克岩的成矿专属性由上地幔成矿元素分布特征决定。与俯冲有关的埃达克质岩浆之所以有利于成矿,重要的原因是存在大量来自俯冲板片的高压、高温流体以及俯冲板片熔融形成高氧逸度(fO2)的熔体,但产出在大陆板内背景、与俯冲无关的埃达克岩的成矿机制还不清楚。根据现代资源勘查理念,结合综合地质信息分析,埃达克质岩具有实际找矿意义。

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