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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。

It was the turn next of the other new arrival, Domoraud, to get in the action when, seizing on a flicked header from Matt Pattison, he strode through on the Woking left to test the Lewes keeper with a left-footed strike. On the quarter hour mark Danny Bunce tried an ambitious effort from 30 yards, the ball passing a foot or so over the bar. A minute later a good move involving Domoraud and Lambu ended in a scramble in the box. The ball reared up but wouldn't come down quickly enough to allow Gez Sole to get the ball on target.

接下来轮到了另一个新加盟的队员，多莫劳德接马特-帕蒂森的头球传球在左路起左脚射门被路易斯队门将化解。15分钟时丹尼-邦斯有一次30米外的劲射，球稍稍高出横梁。1分钟后多莫劳德和朗布在禁区内的配合制造了威胁，但球下坠速度不够，朱塞佩-索勒没能射中目标。

The study showed the recombinant wt/mCREG protein depressed the VSMC proliferation depending on dose and the optimal concentration was 400nM;2biologic function of CREG protein and the membrance receptor mechanism:①effect on VSMC migration: the wound healing experiment showed the OB2 cells migration was slower significantly after added wt/mCREG(400Nm) in supernatant. The HITASY cells migration were very slowly and no remarkable change. The gelatinase digestion and Western blot analysis showed the matrix metalloproteinase was decreased and TIMPs was increased;②effect on differentiation: after added wt/mCREG(400nM), the expression of myocardin, SMα-actin, MHC and caldesmin were increased and that of LM-1 and FN were decreased in OB2 cells. These effects were more significant when adding wtCREG.;③effect on VSMC proliferation: Cell cycle assay and BrDU stain showed: after added the wtCREG and mCREG protein, the ratio of cell in G0/G1 phase increased to 0.5773 and 0.5572 from 0.5308 respectively in OB2 group, which increased to 0.7369 and 0.7034 respectively from 0.6297 in HITASY group;3Role of M6P/IGF2R in CREG biologic function:①ELISA and co-immunoprecipitation showed the wt/mCREG binding to M6P/IGF2R directly.②antibody blocking test: when the anti-IGF2R was added to medium at the same time with wt/mCREG at different concentration(2μg/mL、4μg/mL、8μg/mL),the effects of CREG protein which depressing proliferation, migration, secretion and promoting differentiation were blocked, which had the positive correlation to the concentration of added anti body. The studies showed two combinant CREG promoted VSMC switch to differentiation phaenotype, at the same time, depress VSMC proliferation, migration and secreting extracellular matrix.

上述实验结果证实：两种重组CREG蛋白对VSMC增殖均有剂量依赖性的抑制作用，并且相同浓度的糖基化的CREG蛋白对细胞增殖的抑制效应更为显著，最佳效应浓度为400nM；2两种重组CREG蛋白添加后对HITASY和OB2细胞生物学行为的影响：①CREG蛋白对VSMC迁移的影响：刮伤实验发现，加入最佳效应浓度的wtCREG和mCREG蛋白24h后，OB2组迁移能力下降，HITASY组无明显变化；细胞外基质金属蛋白酶-2，9(Matrix metallo-proteinase 2，9，MMP2 ，9)明胶酶电泳检测和Western blot检测结果证实，两种CREG蛋白均可以使OB2细胞合成细胞外基质MMP2，9减少，而组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases，TIMPs)增加；②CREG蛋白对VSMC分化的影响：加入400nM的wtCREG和mCREG蛋白12h后，OB2细胞myocardin、SMα-actin、MHC、caldesmin表达增加，LM-1、FN表达减少；③流式细胞仪分析细胞周期和BrDU染色分析证实，加入400nM的wtCREG和mCREG蛋白后，OB2组G0/G1期细胞由0.5308分别增加至0.5773和0.5572，HITASY组G0/G1期细胞由0.6297分别增加至0.7369和0.7034；3M6P/IGF2R在重组CREG蛋白的生物学功能中的调控作用：①免疫共沉淀和免疫荧光双染色分析结果显示，CREG蛋白与M6P/IGF2R存在直接结合；②应用抗体阻断实验：将不同浓度的anti- M6P/IGF2R(2、4、8μg /mL)与两种CREG蛋白同时加入培养液中，CREG蛋白抑制VSMC增值、迁移和合成细胞外基质、促进分化的效应减弱，而且与加入anti- M6P/IGF2R浓度正相关。

Part II The experiment study on anti-tumour effect of dendritic cells derived from peripheral blood monocyte of patients with oral cancer after pulsed by tumour antigen in different ways. Collected 50ml peripheral blood from each patients and volunteers to induced DC in vitro. We parted the test group and control group into three subgroups respectively, untreated group A, freezed and thawn group B, RNA transfected group C. To group A, after induced DCs for five days , we added 200U/mlTNF-α in each hole and cultured on. To group B, we added protein antigen (DC/ tumor cells = 1 : 1 ,each hole) for 4h at 5d, and then added 200U/mlTNF-α in each hole and cultured on. To group C, transfected immature DC with tumor cell RNA(DC:RNA=1×105/ml: 5ug)at 5d, operated completely with the specification of invitrogen DMRIE-C reagents. After transfection we replaced transfection agent with complete medium, and then added 200U/mlTNF-α in each hole and cultured on.

第二部分口腔癌患者外周血来源的树突状细胞体外负载抗原后抑瘤作用的实验研究无菌采集20例口腔癌患者和20例健康志愿者外周血50ml，分离单个核细胞体外诱导培养DC，并将实验组和对照组细胞各分为三组，A组为未处理组，B组为冻融组，C组为RNA转染组。A组DC在培养5d后，每孔加入TNF-α200U/ml；B组DC培养第5d后，每孔中加入癌细胞蛋白抗原，使DC/肿瘤细胞=1：1，继续培养4h后，每孔加入TNF-α200U/ml；C组将培养至第5d的未成熟DC用肿瘤细胞RNA进行转染(DC：RNA=1×105/ml： 5ug，按照invitrogen DMRIE-C试剂说明书进行操作)，转染结束后用完全培基取代转染剂，每孔加入TNF-α200U/ml。

Test results are as follows: Ballastless track structures can effectively improve the dynamic action of train loads on subgrade beds, the measured stresses and acceleration rates are far smaller than those measured with ballasted track structures; axle loads have remarkable influence on dynamic stresses of ballastless track subgrades and have some influence on acceleration responses; speeds of either motor car units or freight trains all have limited effect on dynamic embankment responses; after attenuating over 3 meters high embankment, neither dynamic stresses nor acceleration rates are influential to net cushions at the bottom of column-net structure subgrades

结果表明：采用无砟轨道结构可以有效改善列车荷载对路基基床的动力作用，测得的动应力与加速度值均远小于有砟轨道结构测得的值；列车轴重对无砟轨道路基的动应力影响明显，对加速度响应也有一定影响；无论是动车组还是货物列车，其运行速度对路堤部分的动响应影响均有限；动应力与加速度经3 m 高的路堤后衰减，对桩网结构路基下部的网垫层已基本无影响。

The use of multiple regression statistical analysis of past resettlement of academic achievement, proposed to the examination results and school organizational placement tests prorating method of calculation for the future provide the basis for placement of academic computing, bring it closer to the real student academic level; make full use of students in school to participate in the history of the various test results data, follow-up analysis of our students grew up, give full play to our school-based management advantages of large classes in small classes, guiding students in active academic achievement of self-attribution, searching for their own learning methods to help students develop good study of psychological quality; use RANK function analysis of the end of the academic results of students balanced, using radar chart, combined with the median shows the median and the Analysis of 100 students in the end of academic performance, on the one hand to guide students to focus on their overall academic development, and on the other hand, for students in high-rise 2 1 Division of Arts and to provide selected on the basis; to provide easy-to-EXCEL template to enable teachers to gather in peacetime operations or unit tests in the diagnosis of academic information, so that the teaching timely and detailed feedback loops are conducive to Teacher students to keep abreast of the situation, according to the actual situation of students to adjust teaching objectives and teaching strategies in order to better complete the teaching tasks.

利用多元回归统计分析以往的安置性学业成绩，提出以中考成绩与学校组织分班考试按比例折算的计算方法，为今后的安置性学业成绩计算提供依据，使其更接近学生真实的学业水平；充分利用学生在校参加各次测试的历史成绩数据，跟踪分析学生的学业成长历程，充分发挥我校大班小班化管理模式优势，指导学生对自我学业成就进行积极归因，寻找适合自己的学习方法，从而帮助学生形成良好的学习心理品质；利用RANK函数分析学生终结性学业成绩的均衡性，利用雷达图，结合中位数和百位数图示化分析学生终结性学业成绩，一方面引导学生关注自身全面的学业发展，另一方面，为学生在高一升高二文理分科时提供选择依据；提供操作简单的EXCEL模板，方便教师收集平时作业或单元测验中的诊断性学业成绩信息，使教学反馈环节及时精细，有利于任课教师及时了解学生的学习状况，根据学生实际情况调整教学目标和教学策略，从而更好地完成教学任务。

In this experiment,X-ray diffraction apparatus was used forthe first time to examine and research on microdensity,crystallinity and microfibril angle.Through research on the radial variation of bamboo microdensity,this paper points out the conceptions of bamboo hull density and bamboo pulpdensity,and this provides basis for the multiple layer utilization of bamboo.Using computer image-processing technology to carry on precise examinationvolume fraction of bundle sheath,that is key factor to reveal the relationshipbetween bamboo structure and properties.For the first time using resonantvibration means to test the dynamic elastic module,the result shows very ideal.And for the first time researched on the variability of electric conductivityproperty of bamboo.

在本次实验中，首次使用X射线衍射仪对竹材的微密度、结晶度、微纤丝角进行测定和研究，深入研究了竹材微密度的径向变异规律，首次提出竹青密度和竹肉密度的概念，为竹材的径向分层利用提供依据；首次使用微机图象处理技术对竹材的组织比量，主要是纤维束比量进行了精确测定，从而为揭示竹材结构与材性关系打下基础；首次使用共振法对竹材的动态弹性模量进行测定和研究，测量结果是比较理想的；首次研究了竹材导电性的变异。

Measure the items on before and after the two courses of treatment respectively as follows:① Syndromes Investigation: to adopt the scoring methods of symptoms and signs with the certain standard scoring table of symptoms and classifying table of signs for the patients, on the day before and after the two courses of treatment respectively ,② Signs Investigation: the mucoma visibility, swelling or hyperemia conditions, or gloyp and polypoid change in the inferior and middle nasal conchas, nasal septum, etc with a nasal endoscope on the day before and after the two courses of treatment respectively,③ Test of the lung functiaon on the previous day before and after the two courses of treatment respectively.

治疗组在治疗前、两个疗程结束后分别进行以下项目的观察：①临床症状的评定：治疗前及两个疗程结束后当天对患者鼻塞、鼻痒、流涕、喷嚏症状的程度进行评分；②体征的评定：治疗前及两个疗程结束后当天采用鼻额镜肉眼观察患者下鼻甲鼻粘膜颜色、肿胀情况，及中鼻甲是否可见，有否息肉样变等；③治疗前及两个疗程结束后即刻予以检测肺功能。

Measure the items on before and after the two courses of treatment respectively (one course for 15 days) as follows:①Syndromes Investigation: to adopt the scoring methods of symptoms and signs with the certain standard scoring table of symptoms and classifying table of signs for the patients, on the day before and after the two courses of treatment respectively,②Signs Investigation: the mucoma visibility, swelling or hyperemia conditions, or gloyp and polypoid change in the inferior and middle nasal conchas, nasal septum, etc with a nasal endoscope on the day before and after the two courses of treatment respectively,③Test of the interferon-γ、interleukine-4(IL-4) in serum of patients by the method of enzyme linked immunosorbent assay on the previous day before and after the two courses of treatment respectively.

治疗组在治疗前、两个疗程结束后分别进行以下项目的观察：①临床症状的评定：治疗前及两个疗程结束后当天对患者鼻痒、鼻塞、流涕、喷嚏症状的程度进行评分；②体征的评定：治疗前及两个疗程结束后当天采用鼻额镜肉眼观察患者下鼻甲鼻粘膜颜色、肿胀情况，及中鼻甲是否可见，有否息肉样变等；③治疗前及两个疗程结束后一天清晨空腹抽取患者肘静脉血4ml离心后，采用酶联免疫吸附试验检测患者血清IFN-γ、IL-4的含量。

Directed against the problem on confirming vertical bearing capacity of large diameter cast-in-place pile in the west of coteau area of Yunnan province, the paper, analyses the influence degree of the variety of soil physics and mechanics property in area geology condition on bearing capacity of pile foundation, through 5 piles' loading test; the influence degree of construction technics on bearing capacity of pile foundation under the influence of every exterior factors; the influence degree of grouting injection around the pile shaft on bearing capacity of pile foundation.

本文针对滇西山区大直径灌注桩竖向承载力确定存在的问题，进行了5根试桩的静载荷试验，分析了区域地质条件下土的物理、力学性质的变化对桥梁桩基承载力的影响程度；地下水对桩承载力的影响程度；各种外部因素影响下，桩的施工工艺对桩承载力的影响程度；通过在桩体周围土中注浆对桩承载力的影响程度。

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