正常小鼠小肠微皱褶细胞的形态学观察

目的 观察正常小鼠小肠Peyer‘s集合淋巴小结滤泡相关上皮中微皱褶细胞的形态结构。分布及其表面特性。方法 常规的扫描电镜,透射电镜和免疫荧光,免疫冷冻超薄切片技术,结果 微皱褶细胞的粘膜面具有许多短而不规则的微绒毛和微皱褶,基部胞膜向顶部呈穹隆状突起,形成“口袋”样结构,其内含有B、T淋巴细胞和少量的巨噬细胞,顶部胞质内终末网不发达,有许多小泡;荆豆凝集素UEA-1可特异性地与微皱褶细胞结合。结论 微皱褶细胞是分布于Peyer‘s集合淋巴小结圆顶区表面滤泡相关上皮中的一种特化的上皮细胞,利用标记的凝集素UEA-1可作为鉴别微皱褶细胞的依据。     

离体培养细胞的力学实验方法

细胞力学是组织工程和细胞工程的基础之一，在细胞离体培养过程中对细胞施加不同的机械力以研究应力对细胞的影响是细胞力学的一个重要研究领域， 也是细胞力学的重要研究手段。本文综述了离体培养细胞的力学实验方法，根据不同的载荷施加方式可分为：微吸管吸吮技术、压力载荷法、基底拉伸法和流体剪切法等，并指出了各种方法存在的利与弊。     

前列腺基底细胞癌：29例临床病理分析

前列腺基底细胞增生性病变发生于移行区，表现为从基底细胞增生至基底细胞癌的增生性病变谱系。基底细胞增生有宽泛的形态学结构。由基底细胞组成的恶性肿瘤如果与涎腺的对应肿瘤有相同组织学特点，则被命名为“腺样囊性癌”或“腺样基底细胞肿瘤”；大多数缺乏这种相似性，多称为“基底细胞样癌”或“基底细胞癌”。作者研究了29例前列腺基底细胞癌，其中包括前列腺腺样囊性癌的病例。研究的目的是寻找基底细胞癌的形态学谱系的所有特点，以及免疫组化表型和长期生物学行为。     

医用热硫化硅橡胶二段硫化工艺的生物学效应

为考察二段硫化工艺过程对硅橡胶生物学性能的影响,作者根据医用热硫化甲基乙烯基硅橡胶弹性体配方和一定的工艺要求,制取一系列不同二段硫化程度的硅橡胶试样,运用组织培养技术对硅橡胶试样进行生物学测试,特别是测定细胞相对增殖度;进一步利用倒置显微镜观察了相同试样直接接触细胞的形态。结果显示,随着二段硫化时间的增加,细胞相对增殖度增加,而毒性级别降低。     

涎腺基底细胞腺瘤和腺癌与腺样囊性癌的比较观察

目的 研究涎腺基底细胞腺瘤和基底细胞腺癌与腺样囊性癌的形态学特征、免疫表型和鉴别诊断。方法 对5例基底细胞腺瘤，5例基底细胞腺癌和7例腺样囊性癌进行了免疫组化和双重免疫电镜(2例)的观察。结果 基底细胞腺瘤和基底细胞腺癌在免疫表型方面非常相似；基底细胞腺瘤(癌)与腺样囊性癌之间在免疫表型和超微结构方面有一定的差别。结论 基底细胞腺瘤和基底细胞腺癌的鉴别诊断基于两者的生长方式和组织学特征。免疫组化和免疫电镜观察有助于基底细胞腺瘤(癌)与腺样囊性癌的鉴别诊断。     

前列腺基底细胞癌

前列腺基底细胞增生来自于移行区，形态学变化可从基底细胞增生到基底细胞癌。前列腺基底细胞癌比较少见，作者研究了29例基底细胞癌，病人年龄42～89岁（平均69岁）。组织学检查84％（24／29）病例为多种组织学构型，其中腺样囊样构型（AC—P）和小的实性巢伴巢周围细胞栅栏状排列是最主要的构型，各占64％。其他构型包括基底细胞增生样构型9例、小管状构型，部分管内侧偶见玻璃变的胶原纤维9例，其中4例混杂有条索状结构、大的实性巢8例，其中5例中央有坏死。[第一段]     

高分子量细胞角蛋白在前列腺腺癌鉴别诊断中的应用

为提高对各种前列腺增生性病变与癌的认识和鉴别诊断水平，用抗高分子量的细胞角蛋白单克隆抗体３４ＢＥ１２做免疫组织化学染色特异地显示前列腺的基底细胞，并对前列腺腺癌及前列腺增生性病变进行了鉴别，其中前列腺腺癌２１例、前列腺上皮内肿瘤３０例、前列腺非典型性腺瘤样增生５例、前列腺基底细胞增生８例、前列腺萎缩１１例、萎缩后增生４例、前列腺筛状增生３例。结果显示前列腺腺癌基底细胞均丢失，而前列腺增生性病变基底细胞绝大多数保存完好，只少数３级前列腺上皮内肿瘤和非典型性腺瘤样增生基底细胞不完整。研究表明特异地显示基底细胞的高分子量细胞角蛋白３４ＢＥ１２在前列腺腺癌鉴别诊断中有应用价值。     

二维分形维数在皮肤癌光学相干断层成像技术诊断中的应用

目的:在利用光学相干断层成像技术检测和诊断皮肤癌时,获取能够用于皮肤癌诊断的图像特征值。方法:应用差分盒子技术方法,提取色素痣、基底细胞癌和黑色素瘤的光学相干断层图像的二维分形维数。结果:从光学相干断层图像中提取出的二维分形维数能够有效地区分色素痣、基底细胞癌及黑色素瘤,色素痣和基底细胞癌的分形维数较黑色素痣小,具有统计学意义。结论:应用本方法计算的二维分形维数可以用于皮肤癌的诊断。     

细胞骨架在回肠上皮的表达

为探讨细胞骨架在回肠上皮和抗原呈递细胞的表达，用细胞骨架的单克隆抗体对日本正常雌性兔的回肠石蜡切片进行免疫组织化学ＡＢＣ法染色。结果表明，波形蛋白（ｖｉｍｅｎｔｉｎ）表达于回肠淋巴滤泡上皮中的微皱褶细胞（Ｍ细胞）和血管内皮；微管蛋白（α－ｔｕｂｕｌｉｎ）表达于Ｍ细胞、柱状吸收细胞和血管内皮；肌动蛋白（ａｃｔｉｎ）表达于Ｍ细胞、柱状吸收细胞和血管内皮和平滑肌、结缔组织的一些细胞；细胞角蛋白－１８（ｃｙｔｏｋｅｒａｔｉｎ－１８）表达于肠绒毛上皮的纹状缘。本文对细胞骨架的意义进行了讨论。     

神经干细胞在老年性痴呆模型鼠基底前脑内的成活和迁移

为探讨神经干细胞移植入老年性痴呆模型鼠基底前脑的成活和迁移情况,本研究利用无血清培养技术,加表皮生长因子(EGF)和碱性成纤维生长因子(FGF-2)刺激生长,在体外进行神经干细胞的克隆培养,用5-溴-2-脱氧尿苷(BrdU)标记,采用免疫组化和免疫荧光法研究神经干细胞的增殖特性和多向分化潜能,用免疫组化和免疫荧光双标技术观察神经干细胞移植后在老年性痴呆模型鼠基底前脑的成活和迁移。结果显示:克隆细胞球呈nestin和BrdU阳性反应。免疫荧光双标检测,贴壁的细胞为BrdU+GFAP阳性反应或者BrdU+NF阳性反应。移植后3周、4周,移植的针道附近以及整个基底前脑散在有许多BrdU+NF和BrdU+GFAP免疫荧光双标细胞。本研究结果提示,基底前脑神经干细胞能够分化为神经元和星形胶质细胞;基底前脑神经干细胞移植后能够在老年性痴呆模型鼠基底前脑内存活和迁移,可以与脑组织整合,并且能够分化成为神经元和星形胶质细胞。     

Generation of cell-derived three dimensional extracellular matrix substrates from two dimensional endothelial cell cultures

Within the cellular microenvironment, extracellular matrix (ECM) proteins are critical nonsoluble signaling factors that modulate cell attachment, migration, proliferation, and differentiation. We have developed a simple method to isolate and process ECM from endothelial cell cultures to create a three-dimensional (3D) ECM substrate. Endothelial cell monolayers were chemically lysed and enzymatically digested to isolate a thin, two-dimensional (2D) ECM substrate. This thin 1.8 μm 2D ECM was collected and applied to a solid support to produce 12-16-fold thicker 3D ECM substrates with average thicknesses ranging from 21 to 29 μm. The biological activity of isolated ECM was assessed by cell culture. Neural progenitor cells were cultured on endothelial-produced ECM, and unlike the thin 2D ECM, which was quickly remodeled by cells, 3D ECM substrates remained in culture for an extended period (>7 days), suggesting that a continuous signaling cue for in vitro experiments may be provided. This simple method for creating 3D ECM substrates can be applied to a variety of cell culture models for studies aimed at identifying the signaling effects of the ECM within cellular microenvironments.     

实时荧光定量PCR法筛查唐氏综合征的实验研究

目的建立一种无创性、高特异性的快速筛查唐氏综合征（Down’s syndrome，DS）的检测方法。方法采用实时荧光定量PCR技术对正常人和DS患者外周血样本的有核细胞进行检测，根据△Ct值的差异建立检测方法。结果正常组和唐氏组△Ct值有显著性差异（P〈0．001），初步建立了实时荧光定量PCR筛查唐氏综合征的快速检测方法。结论实时荧光定量PCR具有无创性、特异性高且简单、快速等优点，适用于唐氏综合征产前筛查。     

A Rapid Separation Procedure to Analyse Phosphorylation of Two Putative Protein Kinase C Substrates in Intact Lymphocytes

Activation of protein kinase C (PKC) has been implicated in the regulation of many biological processes including the regulation of T and B lymphocyte responses. A direct means of studying activation of this kinase is to analyse specific phosphorylation events of cellular substrates. In the present report we describe a fractionation method that allows quantitative analysis of phosphorylation of two distinct PKC substrates using normal human T cells and T-cell tumour lines. This method, which selectively visualizes PKC-dependent phosphorylation of both an 80- and a 19-kDa substrate, involves three simple fractionation steps and allows a large number of samples to be analysed simultaneously. Since specific antibodies to these cellular substrates are not commonly available, the present method provides an alternative approach which makes it feasible to use phosphorylation of the 19- and 80-kDa proteins as a sensitive marker for PKC activation. Finally, in a cellular system where PKC-mediated phosphorylation of these substrates can be studied without prior purification, the present method results in greatly improved resolution of these phosphorylation events.     

Crystallization kinetics of natural rubber under stress

In the present paper the crystallization kinetics of two networks of natural rubber was analyzed. The two networks were prepared following two different vulcanization techniques, as usual the first, and under swelling conditions the second. The results are discussed at the molecular level and give some important information about the well known deviations from the Gaussian theory on rubber elasticity.     

一种用于医学图像配准的图像外边界的提取方法

医学图像配准是医学图像融合的第一步,而基于图像边界的配准方法对刚性物体图像（如人头颅骨的CT、MRI图像）来讲是一种较好的配准方法。本文给出了一种医学图像外边界的提取方法,它能够快速、准确地提取出CT、MRI等图像的外边界,为用边界法进行图像配准提供了保证。     

基于互信息的多光谱图像配准

研究了多模态医学图像配准的一种，即多光谱图像的配准，分析了该配准存在的困难：运算量巨大，速度较慢，占用内存多，提出了用parzen窗口函数来估计概率密度，以及用样本平均来估计熵；在搜索策略上采用了快速有效的模拟退火算法。实验证明，本文的方法很好地解决了多模态配准中存在的问题，能够快速稳定地实现多光谱图像的配准。     

Laser-induced crystallization of calcium phosphate coatings on polyethylene (PE)

Calcium phosphate (CaP) coatings are used for obtaining a desired biological response. Usually, CaP coatings on metallic substrates are crystallized by annealing at temperatures of at least 400-600 degrees C. For polymeric substrates, this annealing is not possible due to the low melting temperatures. In this work, we present a more suitable method for obtaining crystalline coatings on polymeric substrates, namely laser crystallization. We were successful in obtaining hydroxyapatite coatings on polyethylene. Because of the UV transmission characteristics of the CaP coatings, the use of a low wavelength (157 nm) F(2) laser was necessary for this. As a result of the laser treatment, the CaP coating broke up into islands. The cracks between the islands became larger and the surface became porous with increasing laser energy. The mechanism behind the formation of this morphology did not become clear. However, the fact that crystalline CaP coatings can be obtained on polymeric substrates in an easy way, possibly allows for the development of new products.     

Application of a peptide-based assay to characterize inhibitors targeting protein kinases from yeast

Chemical molecules that inhibit protein kinase activity are important tools to assess the functions of protein kinases in living cells. To develop, test and characterize novel inhibitors, a convenient and reproducible kinase assay is of importance. Here, we applied a biotinylated peptide-based method to assess adenosine triphosphate-competitive inhibitors that target the yeast kinases Hog1, Elm1 and Elm1-as. The peptide substrates contained 13 amino acids, encompassing the consensus sequence surrounding the phosphorylation site. To test whether the lack of distal sites affects inhibitor efficacy, we compared the peptide-based assay with an assay using full-length protein as substrate. Similar inhibitor efficiencies were obtained irrespective of whether peptide or full-length protein was used as kinase substrates. Thus, we demonstrate that the peptide substrates used previously (Dinér et al. in PLoS One 6(5):e20012, 2011) give accurate results compared with protein substrates. We also show that the peptide-based method is suitable for selectivity assays and for inhibitor screening. The use of biotinylated peptide substrates provides a simple and reliable assay for protein kinase inhibitor characterization. The utility of this approach is discussed.     

A substrate-independent method for surface grafting polymer layers by atom transfer radical polymerization: reduction of protein adsorption

A general method for producing low-fouling biomaterials on any surface by surface-initiated grafting of polymer brushes is presented. Our procedure uses radiofrequency glow discharge thin film deposition followed by macro-initiator coupling and then surface-initiated atom transfer radical polymerization (SI-ATRP) to prepare neutral polymer brushes on planar substrates. Coatings were produced on substrates with variable interfacial composition and mechanical properties such as hard inorganic/metal substrates (silicon and gold) or flexible (perfluorinated poly(ethylene-co-propylene) film) and rigid (microtitre plates) polymeric materials. First, surfaces were functionalized via deposition of an allylamine plasma polymer thin film followed by covalent coupling of a macro-initiator composed partly of ATRP initiator groups. Successful grafting of a hydrophilic polymer layer was achieved by SI-ATRP of N,N'-dimethylacrylamide in aqueous media at room temperature. We exemplified how this method could be used to create surface coatings with significantly reduced protein adsorption on different material substrates. Protein binding experiments using labelled human serum albumin on grafted materials resulted in quantitative evidence for low-fouling compared to control surfaces.