小鼠

Methods Study the effect of G. lucidum spores oil (at the dose of 2.5,1.25,0.725 g/kg) on reducing the toxicity of the Cy chemotherapy and 60Co radiotherapy by detecting the number of the peripheral blood leucocyte, the number of bone marrow karyota;spleen index, thymus index of normal mice and H22-bearing mice by carbon clearance test and serum erythrocytolysin test. Results It showed that G.

方法以正常和 H22荷瘤小鼠为对象，通过检测外周血 WBC数、骨髓有核细胞数、脾脏指数、胸腺指数来观察口服灵芝孢子油(2.5，1.25，0.625 g/kg)对于环磷酰胺化疗及60Co放疗的减毒作用；通过小鼠碳廓清实验、血清溶血素测定实验检测药物对 H22荷瘤小鼠免疫功能的影响。

The acute toxicity andmain organs of pathological section for 30 days were observed. Result 1. Ganoderma lucidum spores oil at 0.20g/kg, 0.40g/kg, 1.20g/kg dose couldsignificantly promote the lymphocyte transformation, raise clearance index numberand phagocytic index number, enhance the activity of NK cells and the level of TNF-α, IL-2 in spleen. Ganoderma lucidum spores oil at high dose and middle dose couldsignificantly enhance the serum erythrocytolysin level.

实验结果 1低、中、高灵芝孢子油组(0.20g／kg，0.40g／kg，1.20g／kg)均可提高小鼠单核-巨噬细胞的吞噬指数及碳廓清指数，促进ConA诱发脾淋巴细胞转化，增强小鼠NK细胞活性，提高脾细胞产生TNF-α和IL-2的能力；高、中剂量灵芝孢子油可提高小鼠的血清溶血素水平。

Methods:①Pharmacodynamics observed the effects of The root and leaf of Panicled Fameflower on swimming time of mouse, and physiological function, IL-2 IgA IgM IgG C3 C4 of rat of asthenia of the spleen, and The leaf of Panicled Fameflower on determinating mininal inhibitory concentration, swelling of mouse ear, capillary vein permeability of mouse skin, granuloma in rat.

①药效学实验：观察土人参根、叶对小鼠游泳时间的影响，以及对&脾虚&大鼠脾脏生理功能，细胞因子IL-2、IgA、IgM、IgG、C3、C4的影响，验证土人参根、叶&健脾&功效；观察土人参叶体外抗菌实验，对二甲苯致小鼠耳廓肿胀的影响，对小鼠皮肤毛细血管通透性增高的影响，以及对大鼠棉球肉芽肿的影响，验证土人参叶&解毒消痈&功效。

Trial of acute toxicity: The mouse were given once the drugs of The root (10g/ml)and leaf (9g/ml) of Panicled Fameflower by irrigating gastric in largest volume (0.3ml/10g).

急性毒性实验：土人参根、叶水煎液浓缩制成的最大浓度(土人参根10g／ml，土人参叶9g／ml)给予小鼠最大容积(0.3ml／10g)一次灌胃，若小鼠死亡，则测定半数致死量LD_(50；若毒性很小，测不出半数致死量：则改用一日内以小鼠能耐受的最大浓度和最大容积，灌胃3次；测定其最大给药量。

METHODS: A model of T cell activation and proliferation was established by stimulated the cells with Con A.T cells were treated with different concentrations of CPT. The expression of CD69, the early marker of CD3(superscript +) T cell activation, was measured by FACS. The proliferation index was determined by carboxyl fluorescin diacetate succinmidyl ester by flow cytometry. The cell-cycle distribution was analyzed by propidium iodide staining.

以ConA刺激小鼠T淋巴细胞，建立小鼠T淋巴细胞活化、增殖的模型，以不同浓度的CPT作用于该模型，流式细胞术检测T细胞早期活化标志CD69分子的表达；以活体染料羧基荧光素乙酰乙酸染色流式细胞术分析CPT在ConA刺激下小鼠淋巴细胞的增殖相关指数；以碘化丙啶染色流式细胞术分析细胞周期的分布情况。

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。

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。

RESULTSWhen the dosage of the Aipingxiao were 13.0, 6.5g/kg by intragastric administration, the inhibition rates was 34.2％, 28.1% in the transplantation with S180 Sarcoma, the inhibition rates were 50.8%, 40.8% with H22 liver cancer, the inhibition rates were 74.2%, 57.0% with Lewis lung cancer, respectively. The survival rate increased significantly in the hydroperitoneum type of transplantation with H22 liver cancer more than model group.

结果］分别以13.0，6.5g/kg剂量的癌平消灌胃给药，对小鼠移植性S180肉瘤增殖的抑制率分别为34.2%，28.1%，对小鼠移植性H22肝癌实体瘤增殖的抑制率分别为50.8%，40.8%，对Lewis肺癌实体瘤增殖的抑制率分别为74.2%，57.0%，H22肝癌腹水型小鼠的生存时间明显延长，与模型组比较均有显著性差异。

Law of TT of 肕 of take along sth to sb determines the hyperplasia that EVO and PTX odd in order to and couplet use cells of pair of small rat S180 restrains rate; to build S180 to cultivate model of tumour small mouse hypodermically, observation EVO and PTX odd in order to and couplet use hypodermic to small rat tumour grow inhibition.

采用MTT法测定EVO和PTX单用以及联用对小鼠S180细胞的增殖抑制率；建立S180皮下种植瘤小鼠模型，观察EVO和PTX单用以及联用对小鼠皮下瘤的生长抑制功能。

The experiments may contribute not only to investigating the mechanism of interaction between ldiotype and anti -idiotype T cells by which immune network are up-regulated, but also to analysing the suppressive effect of anti -idiotypic T cell or antibodies against the antigens on the activated T cells in the allo-responsiveness.

本课题选用小鼠耳后心肌移植模型，用同种抗原特异的活化T淋巴细胞免疫同系小鼠的方法，诱导免疫小鼠同种心肌移植物存活时间的延长，以此探讨免疫网络中独特型-抗独特型相互作用机理，着重分析在同种抗原诱导的免疫应答中，抗独特型T细胞和抗活化T淋巴细胞抗原抗体对同种免疫反应的抑制作用。

It has been put forward that there exists single Ball point and double Ball points on the symmetrical connecting-rod curves of equilateral mechanisms.

从鲍尔点的形成原理出发，分析对称连杆曲线上鲍尔点的产生条件，提出等边机构的对称连杆曲线上有单鲍尔点和双鲍尔点。

The factory affiliated to the Group primarily manufactures multiple-purpose pincers, baking kits, knives, scissors, kitchenware, gardening tools and beauty care kits as well as other hardware tools, the annual production value of which reaches US$ 30 million dollars.

集团所属工厂主要生产多用钳、烤具、刀具、剪刀、厨具、花园工具、美容套等五金产品，年生产总值3000万美元，产品价廉物美、选料上乘、质量保证，深受国内外客户的青睐