谷胱苷肽S转移酶在口腔鳞癌中的表达

目的　研究异型增生程度不同的口腔白斑和不同分级的鳞癌中谷胱苷肽S转移酶π(GST π)的表达 ,探讨GST π在口腔鳞癌发生发展中的作用。　方法　采用免疫S -P法 ,对 5 4例轻、中、重度异型增生 ,4 7例高、中、低分化的口腔鳞癌 ,7例口腔粘膜上皮单纯增生患者组织进行GST π检测。　结果　口腔粘膜上皮单纯增生组织中未见GST π的表达 ,轻、中、重度异型增生病例中的GST π阳性率分别为4 7.8%、5 2 .9%和 6 4 .2 % ,高于单纯增生组 (P <0 .0 1) ;高分化鳞癌GST π阳性率为 6 4 .7% ,中、低分化鳞癌组分别为 2 8.5 %及 2 2 .2 % ,中、低分化鳞癌组表达均低于高分化鳞癌组及异型增生组 (P <0 .0 5 )。　结论　GST π表达的变化与口腔鳞癌早期的发生发展密切相关     

寰枢椎侧块关节运动轨迹数学模型的探讨

应用最小二乘法拟合得到寰枢椎侧块关节运动轨迹方程 ,左半部是 :S(x) =-2 3 .2 42 9-8.663 1x-0 .45 91x2 ;右半部是 :S(x) =-19.683 8+ 7.15 5 4x-0 .3 5 2 6x2 。     

Characterization of gp 50, a major glycoprotein present in rat brain synaptic membranes, with a monoclonal antibody

Several cell lines secreting monoclonal antibodies (Mabs) against a major forebrain synaptic membrane (SM) glycoprotein, gp 50, have been raised. Western blots show that the Mabs react with a polypeptide doublet of Mrs 49 and 45 kDa. These polypeptides exist solely in a concanavalin A (Con A) binding form. Removal of the Con A receptors by digestion with endo-beta-N-acetylglucosaminidase H (endo H) lowers the Mrs of the glycoprotein doublet to 36.5 and 34 kDa. Western blots of 2D polyacrylamide gels indicate that gp 50 exists in several isoforms. Solid phase radioimmunoassay (RIA) and Western blots of brain subcellular fractions show the antigenic material to be concentrated in the SM fraction, but to be present in much lower amounts in synaptic junctions and postsynaptic densities. Gp 50 appears to be brain specific. Regional distribution studies show that it is present in all brain regions but is two-fold concentrated in cerebellum, brainstem and midbrain compared to forebrain. Immunocytochemical studies of several brain regions show that gp 50-like immunoreactivity is neuron specific and is concentrated in selected neuronal species, particularly granule cells. In both cerebellar and hippocampal granule cells gp 50-like immunoreactivity is localized in the perikarya and primary dendrites. Though immunocytochemistry did not show staining of synaptic regions this may be due to masking of the reactive epitope. The results are discussed in terms of the molecular properties of gp 50 and its subcellular localization in brain tissue.     

腰椎节段形态对峡部、小关节应力及小关节接触力影响的有限元分析

目的:探讨腰椎形态结构变化对峡部裂性滑脱及小关节退变的作用和意义。方法:采用改良的“非种子区域分割方法”及非平行“最佳切割平面”等一系列新型计算机辅助设计(CAD)方法精确建立包括椎间盘高度、腰椎小关节角、椎间盘前凸角改变的L4~L5活动节段有限元模型;在2700N轴向压缩载荷条件下,分别对各有限元模型的峡部、小关节应力、小关节接触力以及椎间盘负载进行测试。结果:压缩载荷下,腰椎活动节段峡部、小关节等效应力及小关节接触力随椎间盘高度的减小而减小,随小关节角的增大而增加,随椎间盘前凸角的增加而减小。结论:椎间盘高度、腰椎小关节角、椎间盘前凸角等形态结构变化对腰椎节段有限元模型的峡部、小关节应力及小关节接触力有明显的影响。提示腰椎峡部应力性骨折及小关节退变的发生与椎间盘高度、腰椎小关节角、椎间盘前凸角等解剖形态因素有关。     

人血浆作为真皮支架对自体皮肤增殖的影响：组织学和免疫组织化学试验●☆

背景：组织工程是一个多学科研究的交叉学科，其目标是使人体损伤的组织和器官再生，通过这种假设，几乎所有的动物组织都可以在实验室进行培养。一般的方法是从需要移植的患者身上提取干细胞，在一定的支持条件下允许其生长、增殖、生产为可替换的组织。另一方面，寻找细胞能够互相联结并形成分层结构的合适的支持条件，如基质或支架等非常必要。目前用于烧烫伤治疗的材料有很多种，如胶原，透明质酸、纤维蛋白和聚乳酸及其共聚物。 目的：讨论以新型的生物材料自体血浆为支架对自体成纤维细胞和角质化细胞生长、扩张、增殖的影响。 设计：建立一种真皮再生的方法，将自体成纤维细胞浸于人血浆基质中，排除各种影响样本安全性和排斥反应的问题，应用同样的方法在新的真皮上获取角质化细胞。 时间及地点：实验于2008年在意大利曼多瓦C. Poma医院动物工厂完成。 材料：人角质化细胞和成纤维细胞取自1例58岁乳房切除患者的皮肤碎片。实验得到C. Poma医院独立伦理委员会批准，患者知情同意。 方法：从自体活组织皮肤样本中分离人角质化细胞和成纤维细胞，置于培养瓶中增殖，随后将自体血浆作为皮肤移植形成构造的支架，免疫和免疫组织化学特征显示与正常皮肤相似。 主要观察指标：将血浆样本用甲醛固定，埋入石蜡，苏木精-伊红染色，用显微镜进行细胞计数。所有样本均经免疫组织化学评估。 结果：在血浆基质上获得了多层规则形状的角质化细胞和成纤维细胞，基底膜的形成说明自体血浆是角质化细胞和成纤维细胞的生长、分化、扩展的良好支架，真皮和表皮细胞联结重建皮肤移植与正常皮肤无异。 结论：血浆作为角质化细胞和成纤维细胞分化、扩展的支架表现出良好的性能，同时实验还特别发现成纤维细胞浓缩血浆可以暂时替代皮肤，也是角质化细胞生长的强有力的支架。此外血浆还具有廉价，易于制备等优点。自体角质化细胞和成纤维细胞及自体血浆的应用，可以避免血种类型的免疫排斥反应。这种治疗方法给有慢性缺损并且需要连续移植的患者带来了希望。 doi:10.3969/j.issn.1673-8225.2009.47.001     

Detergent fractionation with subsequent subtractive suppression hybridization as a tool for identifying genes coding for plasma membrane proteins

Abstract:  The identification of tumor-specific proteins located at the plasma membrane is hampered by numerous methodological pitfalls many of which are associated with the post-translational modification of such proteins. Here, we present a new combination of detergent fractionation of cells and of subtractive suppression hybridization (SSH) to gain overexpressed genes coding for membrane-associated or secreted proteins. Fractionation of subcellular components by digitonin allowed sequestering mRNA of the rough Endoplasmatic reticulum and thereby increasing the percentage of sequences coding for membrane-bound proteins. Fractionated mRNAs from the cutaneous T-cell lymphoma (CTCL) cell line HuT78 and from normal peripheral blood monocytes were used for SSH leading to the enrichment of sequences overexpressed in the tumor cells. We identified some 21 overexpressed genes, among them are GPR137B, FAM62A, NOMO1, HSP90, SLIT1, IBP2, CLIF, IRAK and ARC. mRNA expression was tested for selected genes in CTCL cell lines, skin specimens and peripheral blood samples from CTCL patients and healthy donors. Several of the detected sequences are clearly related to cancer, but have not yet been associated with CTCL. qPCR confirmed an enrichment of these mRNAs in the rough endoplasmic reticulum fraction. RT-PCR confirmed the expression of these genes in skin specimens and peripheral blood of CTCL patients. Western blotting verified protein expression of HSP90 and IBP2 in HuT78. GPR137B could be detected by immunohistology in HuT78 and in keratinocytes of dysplastic epidermis, but also in sweat glands of healthy skin. In summary, we developed a new technique, which allows identifying overexpressed genes coding preferentially for membrane-associated proteins.     

猫颈部食管的交感神经支配—HRP法

用5只猫于左侧颈部食管壁内注入 HRP 溶液,通过 HRP 逆行追踪法研究颈部食管的交感神经支配,结果表明:1.长轴突型交感节前神经元直接分布到食管壁内,其标记细胞位于双侧脊髓的胸1～3节段,以胸2节段最多(占标记细胞总数的66.45％),注射侧占优势。标记细胞主要位于中间带外侧核(约占95.02%),其次为侧索、中介核、前角腹后外侧核。其细胞形态不一,以中小型细胞为主(占标记细胞总数的90.4％)。2.支配颈部食管的交感节后神经元主要位于星状神经节(约占61.99％),余者位于双侧颈前、中和2～5胸交感节内、以小细胞最多。     

差式氧化数法——计算有机物氧化还原当量的简捷方法

本文提出的“差式氧化数法”特别适用于有机化学反应方程式的平衡。在未平衡方程式之前即可直接求出有机物的氧化还原当量,然后再平衡方程式。较以前先平衡方程式,再间接计算氧化还原当量的方法简便,快速,易掌握。     

Preparation of leukocyte-poor platelets by filtration

There is evidence that leukocyte contaminating red blood cells and platelet concentrates are responsible for refractoriness to platelet transfusions. The efficacy of a cotton-wool filter to remove leukocytes from red blood cells has been documented previously. The present study was designed to evaluate whether the cotton-wool filters can effectively remove leukocytes from platelet concentrates. Sixty pools of random-donor platelets and single-donor plateletpheresis products were filtered through a cotton-wool filter. The efficacy of filtration was determined by measuring the absolute numbers of leukocytes and platelets and subpopulations of mononuclear cells. The average platelet loss was 8% per pool of random platelets and 10% per plateletpheresis product. The average leukocyte removal was 99% from a pool of random platelets and plateletpheresis concentrates collected by CS-3000 and 90% from plateletpheresis concentrates harvested by single-stage COBE/IBM-2997. The filtration removed 100% of granulocytes, 95% of monocytes, 90% of B-lymphocytes, and 85% of T-lymphocytes. We conclude that filtration through a cotton-wool filter is an efficient and cost-effective method for preparation of leukocyte-poor platelets.     

免疫荧光检测人乳头状瘤病毒新技术

利用过氧化氢的氧化作用参与抗原抗体反应,使细胞的膜通透性增强,利于荧光标记抗体染液渗入,从而更快速、广泛地接触抗原,更有效的抗原抗体的嵌合交联。新技术的应用,增强了染色效果,缩短了实验时间,简化了操作,提高了阳性检出率,使实验准确性提高,为临床确诊、疗效观察及预后提供了可靠依据。