梯度法

Methods: Density gradient centrifugalization and absorption technique wererespectively applied to detach peripheral blood mononuclear cells andperipheral blood lymphocytes,T lymphocytes were got by Nylon postabsorption method, and CD4+T and CD8+T lymphocytes divided with anti- CD4 andanti-CD8 antibody by alexin cytotoxic method.

密度梯度离心法分离外周血单个核细胞，玻璃器皿吸附法分离外周血淋巴细胞，尼龙棉柱法分离 T 淋巴细胞，补体介导的细胞毒法分别制备 CD4+T和 CD8+T 淋巴细胞，采用 T 淋巴细胞与角质形成细胞共培养技术研究银屑病患者外周血 T 细胞对角质形成细胞的影响，免疫组化法检测共培养角质形成细胞 Ki67、c-Myc 及 Bcl-xL 蛋白的表达。

In this thesis, we implement a computational procedure to categorize and classify 3-D objects from their contour under a simple environment. First, we use foreground detection, Canny edge detection and speedy GVF snake to obtain the object's contour. The contour is then used to establish the object's features. Two databases are built using methods which compute similarity by modified aspect-combination algorithm proposed by Cyr and Kimia and new proposed aspect-combination algorithm. By using these two databases, we recognize 3-D objects using four recognition methods and compare their performance. Finally, a conclusion is made that our newly proposed algorithm is better than the modified algorithm proposed by Cyr and Kimia.

在本论文中，对一个在单纯环境下的三维物体，由撷取到辨识出此物体做了一个完整的实现，首先，利用前景侦测结合肯尼边缘侦测法和加速的梯度向量流动态轮廓侦测法，来得到物体轮廓，接著利用此轮廓找寻出物体的特徵，然后配合计算相似度的方法，代入修改后的Cyr and Kimia的外观结合演算法（aspect-combination algorithm）和新提出的外观结合演算法去建资料库，然后，对於三维物体则依据两种不同方法所建出的资料库，利用四种辨识的方法来辨识并比较其结果，最后得出新提出的外观结合演算法是优於修改后的Cyr and Kimia的外观结合演算法的结论。

Results The purity as well as removal rates of egg albumin and foreign protein of virus bulk purified by Sepharose 4FF gel chromatography were slightly higher than those by sucrose density gradient centrifugation.

结果 层析法的纯度、卵清蛋白及杂蛋白去除率略高于蔗糖密度梯度离心法，而蔗糖密度梯度离心法的病毒回收率优于层析法。

In this paper,the Newtonian fluid model was used to describe rheological property of the produced fluid,and motion equation of unsteady flow of Newtonian fluid in eccentric annulus with the inner cylinder reciprocating axially was transformed by means of the method of undetermined coefficients.

采用Newton流体模式描述油井产出液的流变性，利用待定系数法变换Newton流体在内管做轴向往复运动的偏心环空中非定常流的运动方程，建立给定流量下Newton流体在内管做轴向往复运动的偏心环空中非定常流的压力梯度计算公式，并利用有限体积法对压力梯度进行数值计算。

In the last part of this dissertation, we discuss how to model an image is discussed and how to segment an image by EDEK.Zero-Cross principle is a classical concept in signal processing. We generalize the concept to be zero normal curvature after the geometry frame built. Furthermore, we present the explanation to choose the dual parameters in SPMVIF. And we research why the Laplacian gradient superior to edge gradient.

梯度方向二阶导数过零原理是信号处理中的经典结论，在运用微分几何的知识建立SPMVIF的几何框架的同时，将这个结论推广到零-法曲率的形式；另外我们对SPMVIF的双参数选取规则进行了讨论并给出它们的几何意义，并且对SPMVIF中法向矢量选取时拉普拉斯梯度优于边缘图像梯度的结论给予统计信息熵的解释。

A proper k total colouring of a graph G is a colouring to its vertices and edges using k colours such that no two adjacent or incident elements of G may be assigned the same colour.

利用梯度投影与罚函数相结合的技巧，将带不等式和等式约束的优化问题化成一个无约束问题，提出了初始点可任意的求解不等式、等式约束优化问题的摄动梯度投影算法；参数δk取不同的数还可以得到一类梯度投影算法，从而得出了在搜索方向和步长不精确条件下的梯度投影法，保证了在实际应用中更容易实现；在较弱条件下，证明了该算法的全局收敛性。

The stable clones are further identified by RT-PCR and Western blot; 6 MTT assay is used to investigate the effect of ZNRD1 on the cell growth of cells (AGS, SGC7901, MKN28, NIH3T3, GES-1); 7 Soft agar assay is used to investigate the effect of ZNRD1 on the clonality of cells (AGS, MKN28); 8 Nude mice assay is used to investigate the effect of ZNRD1 on the cell growth of gastric cancer cells (AGS, MKN28); 9 Flow cytometry is used to investigate the effect of ZNRD1 on the cell cycle distribution of cells (AGS, MKN28, NIH3T3, GES-1); 10 Flow cytometry is used to investigate the effect of ZNRD1 on the cell apoptosis of cells (AGS, MKN28, NIH3T3); 11 MTT assay is used to investigate the effect of ZNRD1 on the drug sensitivity of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR) in vitro; 12 SRCA is used to investigate the effect of ZNRD1 on the drug sensitivity of gastric cancer cells (SGC7901, SGC7901/VCR) in vivo; 13 Flow cytometry is used to investigate the effect of ZNRD1 on adriamycin accumulation of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR); 14 Transmission electron microscope is used to investigate the effect of ZNRD1 on the sensitivity of SGC7901 cells towards drug-induced apoptosis; 15 Flow cytometry and DNA ladder assay are used to investigate the effect of ZNRD1 on the sensitivity of cells (SGC7901, SGC7901/VCR, HL-60/VCR) towards drug-induced apoptosis; 16 Microarray is used to investigate the profiling of ZNRD1-responsive genes in gastric cancer cells (AGS, MKN28, SGC7901, SGC7901/VCR); 17 RT-PCR and Western blot are used to identify the results of microarray; 18 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of cyclin D1; 19 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of MDR1; 20 Kinase assay is used to investigate the effect of ZNRD1 on the activity of cyclin E-CDK2 kinase; 21 The antisensenucleic acids of p21 is used to inhibit the expression of p21, and flow cytometry is used to investigate the effect of p21 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 22 The antisensenucleic acids of p27 is used to inhibit the expression of p27, and flow cytometry is used to investigate the effect of p27 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 23 Liposome is used to up-regulate the expression of Skp2, and flow cytometry is used to investigate the effect of Skp2 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 24 Western blot is used to investigate the effect of ZNRD1 on the stability of Skp2 and p27 in gastric cancer cells; 25 MVD assay is used to investigate the effect of ZNRD1 on the angiopoietic activity of gastric cancer cells; 26 ELISA is used to investigate the effect of ZNRD1 on the expression of VEGF165 in gastric cancer cells; 27 The roles of DARPP-32 in MDR of gastric cancer cells are investigated using gene transfection, MTT assay, SRCA, flow cytometry and DNA ladder assay.

应用杂交瘤技术制备ZNRD1的首个单克隆抗体；2）利用RT-PCR、Western blot和免疫组化检测ZNRD1在胃癌组织、胃炎组织、正常胃上皮组织、胃癌细胞和正常胃组织上皮细胞中的表达；3）构建ZNRD1的小干扰RNA载体，并测序鉴定；4）利用脂质体将ZNRD1的真核表达载体及其空载体转染胃癌细胞（AGS、SGC7901、MKN28）和小鼠成纤维细胞（NIH3T3），G418筛选后进行鉴定；5）利用脂质体将ZNRD1的小干扰RNA载体及其空载体转染药敏胃癌细胞（SGC7901）、正常胃组织上皮细胞（GES-1）、对长春新碱耐药的胃癌细胞（SGC7901/VCR）、药敏白血病细胞（HL-60）、对长春新碱耐药的白血病细胞（HL-60/VCR），G418筛选后进行鉴定；6）利用MTT实验检测ZNRD1高/低表达对细胞（AGS、SGC7901、MKN28、NIH3T3、GES-1）生长的影响；7）通过软琼脂克隆形成实验检测上调ZNRD1对AGS、MKN28细胞克隆形成能力的影响；8）通过裸鼠成瘤实验检测上调ZNRD1对AGS、MKN28细胞体内成瘤性的影响；9）通过流式细胞仪分析ZNRD1高/低表达对细胞（AGS、MKN28、NIH3T3、GES-1）的细胞周期的影响；10）通过流式细胞仪分析上调ZNRD1对细胞（AGS、MKN28、NIH3T3）的凋亡的影响；11）通过MTT实验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）体外药物敏感性的影响；12）通过肾包膜下移植法检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR）体内药物敏感性的影响；13）通过流式细胞仪分析ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）内阿霉素蓄积和泵出的影响；14）通过透射电镜检测上调ZNRD1对SGC7901细胞凋亡敏感性的影响；15）通过流式细胞仪和DNA梯度试验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60）凋亡敏感性的影响；16）通过基因芯片检测ZNRD1高/低表达对胃癌细胞内基因表达谱的影响；17）利用RT-PCR、Western blot对基因芯片的结果进行鉴定；18）利用报告基因实验检测ZNRD1对cyclin D1的启动子活性的调节作用；19）利用报告基因实验检测ZNRD1高/低表达对MDR1的启动子活性的调节作用；20）利用激酶试验检测ZNRD1对cyclin E-CDK2 激酶活力的影响；21）利用反义核酸技术抑制p21的表达；通过流式细胞仪检测抑制p21对ZNRD1介导的细胞周期阻滞的影响；22）利用反义核酸技术抑制p27的表达；通过流式细胞仪检测抑制p27对ZNRD1介导的细胞周期阻滞的影响；23）利用脂质体转染法上调Skp2的表达；通过流式细胞仪检测上调Skp2对ZNRD1介导的细胞周期阻滞的影响；24）利用Western blot检测ZNRD1对p27和Skp2的蛋白稳定性的影响；25）利用微血管密度实验检测ZNRD1对AGS、MKN28细胞裸鼠移植瘤微血管形成的影响；26）利用ELISA检测ZNRD1对AGS、MKN28细胞培养上清和移植瘤匀浆中VEGF165含量的影响；27）利用脂质体转染法、MTT实验、肾包膜下移植法、流式细胞仪和DNA梯度试验检测新耐药相关分子DARPP-32对细胞（SGC7901、SGC7901/VCR、对阿霉素耐药的胃癌细胞SGC7901/ADR）多药耐药表型的影响；利用脂质体转染法和MTT实验检测下调ZNRD1对DARPP-32介导的胃癌多药耐药的调控作用。

Through designing of composition and structure of the bioactive graded coating,innerstress and its distribution in the coating were analyzed and calculated, the resultsshowed that when composition distribution coefficient n was 1.5, a reasonable stressdistribution could be got, that was at the beginning of deposition the suspension containingrichly BG granules was used so that a rich BG granules layer, a good transitional layerbetween BG layer at the bottom and the coating could be obtained at the titanium alloy side,the bottom of the coating; the stress value near the interface and surface and its character,pressure stress or tensile stress, were decided by the character of its composition itself.Changing composition distribution coefficient n could only change the variation tendency ofstress in the coating, but did not change the stress distribution rule in the coating. Thethinner the coating is, the sharper stress variation in the coating is, which does not mean thatthicker coating is better because the thicker the coating is, the little the permitteddeformation of coating is, so the coating thickness should be thinner, for example, about50μm for bending applications, but for applications only bearing pure shear stress, such asroot of tooth implant, the coating can be thicker little, for instance, about 80～100μm. The study on electrification characteristic and electrophoresis deposition of HAand BG granules in aqueous and non-aqueous solution system found that EPD almost didn'toccur in aqueous solution system. However, because HA granules take position charges inabsolute alcohol, a homogeneous EPD be carried out on the cathode titanium alloy slice, but taking negative charge in absolute alcohol the BG granules not be deposited on the cathode. A guided HA crystallizing, 100～300nm, on surface of the BG granules be realized by metathetical reaction, which cover BG granules with HA microcrystals and make the covered BG granules taking position charges in absolute alcohol, sequentially realize the EPCD of the BG and HA granules on the cathode, so it is feasible to make a titanium alloy/BG/HA bioactive graded coating by making use of EPCD technology. The corrosion experiment of rich boron bioglass coating and plasma spray coating showed that split phase, rich boron and rich silicon phase, occurred during its preparation. In basic medium the corrosion behavior of 〓 BG coating showed uniformity corrosion, the corrosion mostly occurred at rich boron phase area, therefore batch formula design of BGshould avoid the occurring of split phase. The corrosion appearance of plasma spray coatingappeared a non-uniform corrosion, mostly occurred at the edge of the laminated HA moltendrops, and emerged an accelerated corrosion tendency, which will easyly lead to corrosioncrackles extending to the interface and the happening of osmotic interfacial corrosion, thatmay be one of the major reasons leading to the coating cracking-off in the later period. Thetesting results of thermal expansion coefficient of 〓 and 〓BG showed the thermalexpansion coefficient of 〓 BG matched with that of titanium alloy better, and 〓 BG couldsinter with titanium alloy into densification enamel layer at low temperature (720℃).

将Ti6Al4V合金在1000℃下进行真空热处理会降低其力学性能，且合金内的V元素会向表面富集，因此，钛合金真空热处理和表面涂层的烧结温度不能过高，即应低于其相转变点；通过对生物活性梯度涂层的组成和结构的设计，分析和计算了梯度涂层内的应力大小和分布，结果表明：对于本研究，当成分分布系数n=1.5时，可以获得较合理的涂层力学性能，即在沉积开始时，采用富含BG颗粒的悬浮液，以便在钛合金侧获得同底层BG有良好过渡的富BG涂层；梯度涂层界面和表面的应力大小、性质由材料组成本身的性质决定，改变成分分布系数，只能改变涂层内应力变化的趋势；涂层的厚薄不影响涂层内的应力分布规律，但涂层越薄，涂层内的应力变化越快，但这并不意味着涂层越厚越好，因为涂层越厚，涂层允许的变形越小，对于应用于弯曲受力部位的涂层而言，涂层应薄一点为好(50μm)；而对于仅纯受剪切应力的部位，如牙根种植体，涂层可适当加厚(80～100μm)；通过对HA和BG颗粒在水溶液体系和非水溶液体系中的带电特性和电泳沉积的研究发现，它们在水溶液体系中很难发生电沉积；在无水乙醇溶液中，HA颗粒带正电，可在阴极钛合金片上发生均匀的电泳沉积，而BG颗粒则带负电荷；利用复分解反应法，可以制得100～300nm的HA，通过诱导HA在BG颗粒表面结晶，可对BG颗粒进行表面包覆，获得了被HA包覆的BG颗粒，改变了BG颗粒表面的带电特性，使BG和HA颗粒在无水乙醇中均带上正电荷，从而实现了HA和BG颗粒在阴极上的共沉积。

The shear layer vortices dominate in the initial portion of the jet, which are a result of the K-H instability. The crossflow encounters an adverse pressure gradient ahead of the jet and separates to form horseshoe vortices under the pressure downstream, and they develop into the wake. The wake vorticity comes into being in the actions of an adverse pressure gradient and a wall-normal component of vorticity, which originates from the wall boundary and is entrained into the jet. The shear layer vortices encounter adverse pressure gradients in the lee of the jet and break down, leading to the formation of the CVP and the CVP is the most important characteristic of the jet in crossflow.

文中指出：剪切层涡源于射流与横流之间的K-H不稳定性；马蹄形涡的产生是由于射流喷口上游迎风侧的壁面边界层内存在逆压梯度，使流动发生了分离，在下游逆压梯度的挤压作用下，形成了马蹄形涡，马蹄形涡向下游发展，进入尾迹区；在逆压梯度和法向涡生成项的共同作用下产生了尾迹涡，尾迹涡源于壁面边界层，止于射流，并将部分涡量向上输运至射流内部的CVP；CVP是横向紊动射流的重要特征，剪切层涡破裂后，在逆压梯度的作用下CVP形成。

Construction of biologically functional bacterial plasmids in vitro . Proc. Natl. Acad. Sci. USA 70, 3240 – 3244 (1973).^ Patricia S. Thomas, Hybridization of Denatured RNA and Small DNA Fragments Transferred to Nitrocellulose PNAS 1980; 77: 5201-5205 PNAS Keith Roberts, Martin Raff, Bruce Alberts, Peter Walter, Julian Lewis and Alexander Johnson, Molecular Biology of the Cell

一个很典型的例子就是DNA分子大小的测量：在凝胶电泳出现之前，DNA分子大小是用蔗糖密度梯度沉降法来测量的，这一方法费时费力而且花费昂贵；而在梯度沉降法出现之前，使用的是黏度法。

Do you know, i need you to come back

你知道吗，我需要你回来

Yang yinshu、Wang xiangsheng、Li decang,The first discovery of haemaphysalis conicinna.

1〕 杨银书，王祥生，李德昌。安徽省首次发现嗜群血蜱。