免疫测定

Methods 40 healthy adult mice were randomly divided into control group, CTX immune-suppressed group, Fb treated group, Fb combined with CTX treated group. The influences of Fb in NK cell activity were determined by the method of NK mediated cytotoxicity test;the multiplicative capacity of lymphocyte was assayed by lymphocyte transformation rate of T cell,and the mice spleen T lymphoblast multiplication analytical method was used to measure the IL-2 level.

健康成年小鼠40只，随机分为对照组、环磷酰胺免疫抑制组、黄蘑碱溶性多糖活性组分正常小鼠给药组和Fb环磷酰胺伍用给药组，采用NK细胞介导的细胞毒试验测定自然杀伤细胞的活性，淋巴细胞转化试验测定T淋巴细胞增殖功能，鼠脾T淋巴母细胞增殖分析法测定白细胞介素2（IL-2）的活性，观察Fb对各组小鼠免疫指标的影响。

61Healthy persons were selected as healthy group. The serum concentration of TGF-β1 was measured by ELISA method; nitre oxide was measured by nitrit reductase method, plasma Ang II was measured by radioimmunoassay.

对高血压病人和61例健康对照者应用酶联免疫吸附法测定血清TGF-β_1浓度，硝酸还原酶法测定NO的浓度，放射免疫分析法测定Ang Ⅱ水平。

The immune function was evaluated with T lymphocyte proliferation,NK cell activity assay,IgMplaque forming cell assay,blood serum erythrocytolysin assay and delayedtypehypersensitivity to SRBC.

采用T淋巴细胞增殖和迟发型变态反应来评价细胞免疫功能，采用血清溶血素测定和抗体生成细胞检测来评价体液免疫功能，采用乳酸脱氢酶法测定自然杀伤细胞活性来评价非特异性免疫功能。

Cases healthy subjects served as control. In addition, density of glomerular matrix membrane and quantity of cell in glomeruli were measured with CMIAS image analysis system by computer. Global sclerosis, cellular crescents, fiber or fibrocellular crescents were measured with Memphis scope. Tubulointerstitial lesions were examined with semi-quantitative grades (including O-III grades).Results Compared to normal control, there was a significant increase of urinary TGF-betaK IL-6 and Col-lV levels in patients with IgAN (p. 01). There were also significantly positive correlation between the levels of urinary TGF-betaK Col-IV and density of glomerular matrix membrane ., interstitial fibrosisp.

采用酶联免疫吸附法测定尿TGF-β1(Transforming growth factor-β1)、尿IL-6(interleukin-6)和尿Col-Ⅳ(type Ⅳ Collagen)；采用免疫组织化学法检测肾组织TGF-β1、Col-Ⅳ表达；应用CMIAS多功能真彩色病理图像分析系统对肾小球基质基底膜面密度和细胞个数进行半定量测定；对球性硬化、节段硬化、细胞新月体及纤维或纤维细胞新月体所占肾小球百分数和肾小球IgA沉积免疫荧光强弱的判断采用Memphis法进行评分；肾小管间质病变程度采用光镜下半定量分级法(包括0-Ⅲ级)。

The method uses first phase law to determine 63 DVT patient contrasts normally with 30 the F Ⅻ C of the group, use law of thing of the heart that deliver quality to detect at the same time zymogen of serous fine dissolve is active , law of enzymatic couplet immunity measures sex of antigen of content of activation of zymogen of methodize fine dissolve (depressor of content of activation of zymogen of dissolve of T-PAAg), fine - 1 antigen sex (PAI-1Ag) and immunity compare chaotic law to determine serous D-2 gets together body content.

方法采用一期法测定63例DVT患者和30例正常对照组的FⅫC，同时采用发色底物法检测血浆纤溶酶原活性，酶联免疫法测定组织型纤溶酶原激活物抗原性、纤溶酶原激活物抑制剂-1抗原性(PAI-1Ag)及免疫比浊法测定血浆D-二聚体含量。

Image analysis system was used for semi-quantitive assay of TGF-βRⅠ and R Ⅱexpression, and microphotography was used to record the morphologic changes of culturedchondrocytes. The results are:1. TGF-〓 showed sequentially inhibitory and promotive effects on the proliferation of human articular chondrocytes of two week monolayer culture from one passage to eight passage.

本研究采用CellTiter 96〓单溶液细胞增殖分析试剂盒测定体外培养中的软骨细胞的增殖活细胞数量；免疫组化法测定软骨细胞TGF-βR Ⅰ、R Ⅱ，Ⅱ型胶原和S-100蛋白的定性表达；酶免分析法测定TGF-〓作用后的Ⅱ型胶原定量表达；通过图象分析仪，对TGF-βRⅠ、R Ⅱ的免疫组化染色结果进行半定量分析；本研究的实验结果如下：1。

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 objective was to determine the effects of active immunization of T-3-BSA in Rom cockerels, 400 cockerels were divided into 4 groups, group 1,2 and 3 were treated with T-3-BSA on different dose(0.2,0.4,0.8mg/mI respectively),within each group,there were 5 treatments following different vaccination schedules, group 4 served as controls. Every injection of immunogen was conducted on different sites around the neck, and the datas were measured at week 11,15,19, combs,wattles,body weight,serum T concentration,carass traits and testicular development and histology were studied in order to find out the most reasonable vaccination schedule and the most possible mechanism in the H-P-G axis of T.

本试验测定了睾酮—3—牛血清白蛋白(T-3-BSA)主动免疫对公鸡性腺与冠垂发育及雄激素水平的影响。400只罗曼蛋用雏公鸡除随机取25只作对照组外，另375只随机分成3大组，每大组125只再随机分成5小组，每小组25只，3大组分别使用0.2mg/ml、0.4mg/ml和0.8mg/ml的T-3-BSA免疫注射，每大组的①②③小组分别于3、6、9周龄免疫注射，④小组于3周龄首次免疫，6周龄加强免疫，⑤小组于3周龄首次免疫，9周龄加强免疫。

METHODS Dinitrofluorobenzene was used to induce the contact dermatitis in mouse ear, rhIL-11 0.375-1.5 mgkg^(-1)d^(-1) sc for 10 d, the degree of skin inflammatory reaction was observed. At the same time, the serum level of tumor necrosis factor-α and interleukin-6(IL-6) was measured by radioimmunoassay; the serum level of nitrogen monoxidum was detected by biochemical method and the expression of intercellular adhesion molecule-1(ICAM-1) was detected by immunohistochemical method.

采用2，4-二硝基氟苯致小鼠耳部皮肤接触性皮炎试验模型，观察sc rhIL-11 0.375~1.5mgkg^(-1)d^(-1)，连续10d后皮肤炎症反应程度，同时采用放射免疫分析法测定血清中肿瘤坏死因子-α和白细胞介素-6(IL-6)的水平，采用生物化学检测法测定血清一氧化氮的水平，采用免疫组化法测定皮肤中细胞间粘附分子(ICAM-1)的表达。

Breath, muscle contraction of the buttocks; arch body, as far as possible to hold his head, right leg straight towards the ceiling (peg-leg knee in order to avoid muscle tension).

呼气，收缩臀部肌肉；拱起身体，尽量抬起头来，右腿伸直朝向天花板（膝微屈，以避免肌肉紧张）。

The cost of moving grain food products was unchanged from May, but year over year are up 8%.

粮食产品的运输费用与5月份相比没有变化，但却比去年同期高8％。