转移相关基因nm23和P53及S100A4在晚期胃癌中的表达及与侵袭转移的相关性研究

目的探讨转移相关蛋白（nm23和P53及S100A4）在晚期胃癌转移发展过程中的意义。方法应用组织芯片和免疫组织化学方法研究nm23、P53和S100A4在74例胃癌患者的癌、癌旁、淋巴结转移、远处转移（肝脏、胰腺、卵巢等）组织中的表达。结果同癌旁组织相比，癌组织中P53和S100A4表达明显升高（P〈0．01），而nm23的表达明显降低（P〈0．05）；同癌组织相比，淋巴结中nm23的表达明显降低（P〈0．05）。远处转移组织S100A4表达明显升高（P〈0．01）。3者的阳性表达与胃癌的部分恶性生物学行为有关。癌组织中nm23^＋／P53^＋、P53^＋／S100A4^＋和nm23^＋／S100A4^＋的基因表型比例分别为48例（64．9％）、50例（67．6％）和39例（52．7％），其中P53^＋／S100A4^＋、nm23^＋／S100A4^＋和nm23^＋／P53^＋／S100A4^＋均与胃癌的高转移潜能相关。结论nm23和S100A4在肿瘤转移中发挥了重要作用；联合检测nm23、P53和S100A4的表达可用于评定胃癌的转移潜能。     

Microarray analysis of healing rat Achilles tendon: evidence for glutamate signaling mechanisms and embryonic gene expression in healing tendon tissue.

Tendon healing is a complex process consisting of a large number of intricate pathways roughly divided into the phases of inflammation, proliferation, and remodeling. Although these processes have been extensively studied at a variety of levels in recent years, there is still much that remains unknown. This study used microarray analyses to investigate the process at a genetic level in healing rat Achilles tendon at 1, 7, and 21 days postinjury, roughly representing the inflammation, proliferation, and remodeling phases. An interesting temporal expression profile was demonstrated, identifying both known and novel genes and pathways involved in the progression of tendon healing. Both inflammatory response and pro-proliferative genes were shown to be significantly upregulated from 24 h postinjury through to 21 days. Day 7 showed the largest increase in genetic activity, particularly with the expression of collagens and other extracellular matrix genes. Interestingly, there was also evidence of central nervous system-like glutamate-based signaling machinery present in tendon cells, as has recently been shown in bone. This type of signaling mechanism has not previously been shown to exist in tendon. Another novel finding from these analyses is that there appears to be several genes upregulated during healing which have exclusively or primarily been characterized as key modulators of proliferation and patterning during embryonic development. This may suggest that similar pathways are employed in wound healing as in the tightly regulated progression of growth and development in the embryo. These results could be of use in designing novel gene-based therapies to increase the efficacy and efficiency of tendon healing.     

应用抑制消减杂交和cDNA表达谱芯片筛选肝星形细胞持续活化的相关基因

目的：利用抑制消减杂交（suppression subtractive hybridization,SSH）和cDNA表达谱芯片筛选早期活化的肝星状细胞与大鼠正常肝脏组织、轻度肝纤维化组织、重度肝纤维化组织中差异表达的基因.方法：从SSH构建的大鼠轻度和重度肝纤维化中两个差异cDNA文库中挑选1 000条上调显著的基因,与正常大鼠4 136条基因克隆制作成一张芯片,筛选持续期活化的肝星状细胞相关基因.结果：获得与HSC持续活化相关的上调基因633条,下调基因715条,其中血清和糖皮质激素调节蛋白激酶（SGK）在正常大鼠基因克隆和2、8周SSH上调差异基因中均呈上调信号.结论：联合应用SSH和cDNA表达谱芯片是筛选和鉴定不同样本中差异表达基因的快速、经济和有效方法;SGK可能作为多种细胞信号传导通路和细胞磷酸化级联反应的一个功能性交汇点,参与了肝星状细胞的早期活化和信号传导.     

Allergen microarray: comparison of microarray using recombinant allergens with conventional diagnostic methods to detect allergen-specific serum immunoglobulin E

BACKGROUND: The availability of recombinant allergens and recent advances in biochip technology led to the development of a novel test system for the detection of allergen-specific IgE. OBJECTIVE: To test the performance of this allergen microarray in a serological analytical study. METHODS: Standard allergens contained in grass pollen (Phl p 1, Phl p 2, Phl p 5 and Phl p 6) and tree pollen (Bet v 1 and Bet v 2) were used as a model system. The detection of allergen-specific serum IgE using microarrays was compared with standard test systems: CAP/RAST and an in-house ELISA. In order to test the analytical sensitivity of the assays, geometric dilutions of a serum pool containing high levels of pollen-specific IgE from allergic individuals were tested in each system. To assess the analytical specificity, the sera of 51 patients with presumptive allergic symptoms were collected before diagnosis. Thereafter, the results for grass/tree-pollen-specific IgE were compared. RESULTS: The microarray has a good dynamic range similar to the CAP/RAST system. Microarray and ELISA showed comparable analytical sensitivity exceeding the CAP/RAST system. With respect to the analytical specificity, no significant cross-reactivity of the allergens was observed. For two of the allergens tested, weak positive signals were detected in the microarray test system, whereas they were not detectable by CAP/RAST. CONCLUSION: A good correlation of presently used methods to detect serum IgE and the novel microarray test system was observed. As a next step, a careful validation of this method for a multitude of allergens and a thorough clinical evaluation has to be provided. Microarray testing of allergen-specific IgE can be presumed to be the method of choice for a prospective component-resolved diagnosis of Type I allergy, and the basis for the design and monitoring of a patient-tailored specific immunotherapy in the future.     

利用cDNA微阵列技术筛选成年期与胚胎期睾丸差异表达基因——钙联接蛋白基因

目的 :用cDNA微阵列技术克隆和筛选成年期与胚胎期睾丸差异表达基因 ,以进行睾丸发育及精子发生的调控研究。　方法 :构建人睾丸cDNA芯片 ,分别用正常成人及胚胎睾丸的mRNA探针进行杂交 ,对成人和胚胎睾丸中差异表达的基因进行高流量的比较。　结果 :发现 1条成人睾丸高表达、胚胎低表达的基因———钙联接蛋白 (CLGN)基因。　结论 :运用cDNA微阵列方法可筛选到成年期与胚胎期睾丸差异表达基因     

一种用于基因表达谱分析的基因芯片方法

目的：通过研究SLE患者外周血基因表达谱的改变，建立一种新型的基因芯片技术用于分子发病机理的研究。方法：提取总RNA，逆转录合成单链cDNA、双链cDNA，体外转录合成生物标记的cRNA与基因芯片进行杂交，通过抗体的检测标记荧光染料Cy3，基因芯片扫描仪进行图像扫描。结果：该方法有较高的重复性和稳定性，SLE患者与正常对照组相比较，鉴定出94个基因存在表达差异。结论：该方法能在较少起始标本量的情况下，有效地进行SLE致病基因的筛选，更好的理解SLE发生的分子机理，并且可在其它疾病研究中推广应用。     

生物医学测量及控制技术新进展

介绍生物医学测量及控制技术领域中的一些研究进展。     

运用组织芯片技术检测COX-2在结直肠癌中的表达及其临床意义

目的 探讨结直肠癌组织中COX-2的表达及其与各临床病理因素的关系，评价COX-2在结直肠癌预后判断中的价值。方法 应用组织芯片技术结合免疫组织化学SABC法，检测126例早中期结直肠癌组织中COX-2的表达情况，回顾性分析COX-2与各临床病理因素及预后之间的关系。结果 根据免疫组化染色强度，所有病例被分为COX-2高表达组和低表达组，其中高表达组有32例（25．4％），低表达组有94例（74．6％）。COX-2在结直肠癌中表达情况与年龄、性别、肿瘤大小、肿瘤部位、组织学类型、浸润深度、淋巴是否转移、Dukes分期均无相关。然而，高表达的COX-2与肿瘤复发、特别是血行转移显著相关（P〈0．05）。两组之间生存率具有显著差异（P=0．0067），COX-2高表达组术后五年生存率显著低于低表达组。多因素回归模型分析结果显示．在潜在的预后因素中（年龄、性别、肿瘤大小、肿瘤部位、组织学类型、淋巴是否转移、Dukes分期、COX-2表达），COX-2表达和Dukes分期可作为是结直肠癌根治术后独立预后因素，COX-2的检测可作为结直肠癌患者预后判断的一个有价值的指标。结论 高表达的COX-2与肿瘤复发、特别是血行转移显著相关，COX-2的检测町作为结直肠癌患者琐后判断的一个有价值的指标：应用组织芯片高效检测临床组织样本具有快速、方便、经济、准确的优点。     

几种性传播疾病病原体检测芯片的制备

目的 :为了同时多样本检测和鉴别淋病奈瑟球菌、沙眼衣原体和解脲脲支原体 3种重要的性传播疾病病原体 ,制备了寡核苷酸检测芯片。方法 :针对 3种病原体和荧光素酶基因设计特异的引物和寡核苷酸探针 ,采用硫代和氨基双功能探针修饰技术制备寡核苷酸芯片 ,以荧光标记多重不对称PCR技术为基础 ,通过将单链PCR产物与芯片杂交实现对性传播疾病病原体的检测。结果 :对 10种与待检病原体无关的菌及定量有限稀释的荧光素酶和 3种病原体基因质粒模板进行芯片检测 ,结果表明芯片对待检病原体特异 ,其检测 4种基因的灵敏度均为 5×10 3 拷贝质粒。对 2 4份性传播疾病患者标本进行芯片检测 ,沙眼衣原体感染率为 10 0 % ,与淋病奈瑟球菌混合感染率为 83.3% (2 0 / 2 4 ) ,与传统PCR诊断结果完全一致。在 2 4份标本中 ,淋病奈瑟球菌、沙眼衣原体和解脲脲支原体三重感染病例芯片诊断为 3例 ,混合感染率为 12 .5 % (3/ 2 4 ) ;而传统PCR诊断为 4例 ,混合感染率为 16 .7% (4/2 4 ) ,两种方法的符合率为 75 %。结论 :该芯片是一种可靠检测 3种病原体的方法 ,它可快速提供有关患者混合感染的情况 ,因而为指导个性化治疗提供及时可靠的诊断依据。