细胞凋亡相关基因研究进展

细胞凋亡现象由于广泛地存在于个体发生过程及免疫机制中，且与抗癌、抗衰老、防治ＡＩＤＳ等密切相关，因而引起越来越多科学家们的重视，对其的研究也已推进到分子水平。通过分子生物学技术的应用，对细胞凋亡相关基因的研究也不断深入，本文主要综述了近年来细胞凋亡相关基因的研究进展。     

新闻

新闻我国口腔医学技术跻身世界先进行列从在沪召开的首届中国国际口腔医学研讨会上获悉，我国口腔医学的基础研究和临床医疗技术已达到国际先进水平，有些领域已跃居世界前列。据介绍，我国口腔医学领域的研究已广泛应用于生物和计算机技术，进入了细胞和分子水平，逐步与...     

分子工具在寄生虫学研究中的应用

现代分子技术对寄生虫的基础研究和应用研究都有着重要的影响,尤其是PCR相关技术有着更广泛的应用,因为PCR反应能够极其敏感的从少量的寄生虫材料中扩增到所需的DNA片段,另外高分辨率的电泳技术和基因组方法应用也越来越多.本文对核酸技术如聚合酶链式反应、突变扫描技术、指纹技术的发展和应用进行了综述,并着重强调了这些核酸技术在寄生虫方面的用途和潜力.     

PCR技术在分子肿瘤学研究中的应用

聚合酶链反应(PCR)技术的应用发展很快,近年来,已成为分子肿瘤学研究的重要方法。本文综述了 PCR 技术在肿瘤病毒基因、癌基因和抗癌基因等方面研究中的有关进展,并讨论其发展前景。     

PCR分子信标法及其应用

PCR分子信标法是近几年发展起来的一种全新的核酸定量检测技术，它具有PCR技术的高效核酸扩增特性，核酸探针杂交的高特异性，以及光谱技术的高灵敏性和可计量性。目前，PCR分子信标法已被广泛应用于病原微生物的检测、肿瘤研究、基因突变研究等多个领域。     

Clq及与基因工程抗体相互作用的结构功能研究

Ｃｌｑ结构复杂，Ｃｌｑ相关分子的发现和ＣｌｑＲ在多种细胞上的存在，表明ｃｌｑ具有较广泛的生物学功能。利用基因工程技术，对分子作特定重组、缺失、突变，建立重组分子相互作用的结构与功能研究模型，已在Ｃｌｑ和基因工程抗体的研究中取得许多进展。这方面深入探讨具有重要意义。     

Clq及与基因工程抗体相互作用的结构功能研究

Ｃｌｑ结构复杂，Ｃｌｑ相关分子的发现和Ｃｌｑ在多种细胞上的存在，表明Ｃｌｑ具有较广泛的生物学功能。利用基因工程技术，对分子作特定重组、缺失、突变，建立重组分子相互作用的结构与功能研究模型，已在Ｃｌｑ和基因工程抗体的研究中取得许多进展。这方面深入探讨具有重要意义。     

以双向凝胶电泳为基础的食管癌组织蛋白质组学分析新策略

目的：探讨采用双向电泳为基础的蛋白质组学的研究手段在食管癌组织与食管癌旁组织差异分析中的应用。方法：双向凝胶电泳分离食管癌及其癌旁组织的蛋白质，图像分析软件ImageMaster4．01分析所获得的凝胶图谱并寻找差异点，用质谱仪鉴定差异点蛋白质。结果：提出了一套可行的癌组织差异分析策略，并发现在分子量50 000-60 000，等电点5—6的区域，食管癌组织和食管癌旁组织存在2个差异表达的蛋白，质谱鉴定为丙酮酸激酶M2/M1。结论：在合理的策略指导下，以双向凝胶电泳为基础的蛋白质组学技术是研究肿瘤的一种重要手段，我们得到的丙酮酸激酶M2/M1在食管癌与其癌旁组织中存在差异表达，可能对食管癌的诊断、治疗具有重要意义。     

核磁共振技术在现代药理学研究中的应用

核磁共振（ＮＭＲ）技术在生物医学研究领域已得到了广泛的应用，已成为现代生理、生物化学和药理学研究中新的重要手段之一。该技术具有非损伤性，直接测定活体动物、离体器官和组织细胞内多种化合物，可以在生理或病理状态下进行连续动态测定，有利于药物代谢动力学研究。ＮＭＲ技术用于脑、心、肌肉等组织的生理、病理及药物作用测定的报导逐年增加，本文就国内外ＮＭＲ技术与药理学有关的应用综述如下。     

一种逆转录－聚合酶链反应产物的快速分子克隆方法及其在丁型肝炎病毒基因组克隆中的应用

一种逆转录－聚合酶链反应产物的快速分子克隆方法及其在丁型肝炎病毒基因组克隆中的应用刘善虑，詹美云（中国预防医学科学院病毒学研究所，北京１０００５２）聚合酶链反应（ＰＣＲ）作为一种高效的ＤＮＡ扩增技术在当今分子生物学研究领域已得到广泛的应用；尤其是在逆...     

Tissue microarray (TMA) technology: miniaturized pathology archives for high-throughput in situ studies

Tissue microarray (TMA) technology allows a massive acceleration of studies correlating molecular in situ findings with clinico-pathological information. In this technique, cylindrical tissue samples are taken from up to 1000 different archival tissue blocks and subsequently placed into one empty 'recipient' paraffin block. Sections from TMA blocks can be used for all different types of in situ tissue analyses including immunohistochemistry and in situ hybridization. Multiple studies have demonstrated that findings obtained on TMAs are highly representative of their donor tissues, despite the small size of the individual specimens (diameter 0.6 mm). It is anticipated that TMAs will soon become a widely used tool for all types of tissue-based research. The availability of TMAs containing highly characterized tissues will enable every researcher to perform studies involving thousands of tumours rapidly. Therefore, TMAs will lead to a significant acceleration of the transition of basic research findings into clinical applications.     

Expected usefulness of in situ hybridization for clinical pathology

In situ hybridization technique was developed as molecular-histological usefulness at 1980's. To consider usefulness in the clinical pathology, we should recognize avairabilities of recent techniques such as DNA tip. The characteristic to in situ hybridization is summarized as follows. 1. Diagnosable gene expression pattern with histological feature in the section. 2. Need small amount of sample for hybridization. 3. Easy to distinguish expression of genes that show highly homologies. Previously, we developed non-radioisotopic in situ hybridization by digoxigenin-UTP labeled cRNA probe. The sensitivity and resolution are sufficient for detecting 50 copies mRNA in individual cells. This technique is expected to utilize for molecular diagnosis in clinical field in the near future.     

Controversies in the assessment of HER-2: more questions than answers

Human epidermal growth factor receptor 2 (HER-2) is over-expressed in 15% to 30% of breast cancers and is a poor prognostic marker in node-positive patients. HER-2 expression is an indicator of greater sensitivity to anthracycline-based chemotherapy and is the major criterion for selection for treatment with the anti-HER-2 antibody trastuzumab (Herceptin). Fluorescence in situ hybridization and immunohistochemistry (IHC) are the 2 most commonly used methods for detection of the gene and protein, respectively. Criticisms have been levied at the IHC method of identifying HER-2 overexpression but convenience and costs of this technique cannot be overlooked. Modifications to the IHC technique and scoring accommodate for many of the problems that derive from variables in preanalytical and analytic factors that influence results but standardization is currently impossible to attain. Deficiencies in fluorescence in situ hybridization assay also exist and alternative molecular methods of assay are explored in this review.     

Buffered formalin is the superior fixative for the detection of HPV DNA by in situ hybridization analysis.

In situ hybridization is used commonly for detection of human papillomavirus (HPV) DNA. There is little information, however, on whether the detection of HPV DNA by in situ hybridization can be affected by the way in which the tissue is fixed. To address this question, the authors compared the hybridization signal using this technique under low stringency conditions for several genital condylomata containing HPV 6 or 11 that were randomly subdivided and fixed in various fixatives for 16 hours. In all cases, the largest proportion of cells with koilocytotic atypia that had detectable HPV DNA was in buffered formalin-fixed tissue (80%), followed by tissue fixed in unbuffered formalin (70%), Hartman's solution (40%), and Bouin's solution (10%). After a high stringency wash, the greatest decrease in the overall hybridization signal was with tissue fixed in Bouin's solution; a minimal decrease was noted with tissue fixed in buffered formalin. Fixation in Bouin's solution for 2 hours gave in situ hybridization results comparable with buffered formalin fixation but with poorer cytologic detail. It is concluded that, of the fixatives studied, buffered formalin is superior for the detection of HPV DNA by in situ hybridization analysis.     

Comparison of different techniques for the detection of genetic risk-identifying chromosomal gains and losses in neuroblastoma

Neuroblastoma (NB) is a pediatric neoplasia that shows complex combinations of acquired genetic aberrations. The specific genes and the molecular mechanisms responsible for development and progression of NB remain poorly understood. Our main objective is to compare the results obtained with different techniques for the detection of genomic data in 20 patients with NB using the information obtained to select the appropriate technique in routine analysis for the therapeutic stratification. The genetic methods used in this study are multiprobe fluorescence in situ hybridization (FISH) assay, metaphasic comparative genomic hybridization (mCGH), array comparative genomic hybridization (aCGH), and the multiplex ligation-dependent probe amplification (MLPA). Genomic copy number abnormalities were used to group the cases in four categories: MYCN amplification cases; 11q deletion tumors; cases with partial chromosome gains or losses and samples with entire chromosome alterations. The data obtained from the multigenomic techniques showed a high degree of concordance and our findings support the hypothesis that NB consists of biologically distinct subgroups that differ by genetic characteristics of prognostic relevance. FISH will be essential for the mandatory study of MYCN status. The use of MLPA as routine technique is an advantage procedure for detecting the implication of the common genetic alterations in NB.     

荧光原位杂交技术在染色体病产前诊断中的应用及前景

荧光原位杂交是近年来发展起来的检测染色体异常的新型技术,在产前诊断中也得到了广泛的应用,成为传统细胞遗传学检测的一种重要补充。本文着重介绍了FISH技术的原理及探针的分类,FISH技术在染色体病产前诊断中的应用及前景。     

Tumor cytogenetics revisited: comparative genomic hybridization and spectral karyotyping

Fluorescence in situ hybridization techniques allow the visualization and localization of DNA target sequences on the chromosomal and cellular level and have evolved as exceedingly valuable tools in basic chromosome research and cytogenetic diagnostics. Recent advances in molecular cytogenetic approaches, namely comparative genomic hybridization and spectral karyotyping, now allow tumor genomes to be surveyed for chromosomal aberrations in a single experiment and permit identification of tumor-specific chromosomal aberrations with unprecedented accuracy. Comparative genomic hybridization utilizes the hybridization of differentially labeled tumor and reference DNA to generate a map of DNA copy number changes in tumor genomes. Comparative genomic hybridization is an ideal tool for analyzing chromosomal imbalances in archived tumor material and for examining possible correlations between these findings and tumor phenotypes. Spectral karyotyping is based on the simultaneous hybridization of differentially labeled chromosome painting probes (24 in human), followed by spectral imaging that allows the unique display of all human (and other species) chromosomes in different colors. Spectral karyotyping greatly facilitates the characterization of numerical and structural chromosomal aberrations, therefore improving karyotype analysis considerably. We review these new molecular cytogenetic concepts, describe applications of comparative genomic hybridization and spectral karyotyping for the visualization of chromosomal aberrations as they relate to human malignancies and animal models thereof, and provide evidence that fluorescence in situ hybridization has developed as a robust and reliable technique which justifies its translation to cytogenetic diagnostics.     

Fluorescent in situ hybridization and array comparative genomic hybridization: complementary techniques for genomic evaluation

During the past few years a new high-throughput molecular technology, array comparative genomic hybridization, has received a great deal of attention. As a DNA-based tool, this technique is presumably more reproducible than expression arrays. In this review, I discuss how array comparative genomic hybridization is remarkably similar with regard to genome analysis to fluorescent in situ hybridization, a technique that is generally regarded as one of the more accurate and reproducible molecular techniques in diagnostic surgical pathology. A thorough understanding of this technology will be useful for all surgical pathologists in the near future, as this technology will no doubt have some influence on our daily practice.