基因诊断技术的临床应用进展

 近年来,基因诊断技术取得前所未有的进步,随着该技术在临床检测中的推广、应用,基因诊断技术为我们早期发现肿瘤以及其他遗传性疾病带来曙光。本文拟从基因诊断中常用的分子生物学技术及其在临床中的应用做一总结。     

环介导等温扩增技术及其在病原微生物检测中的应用

 环介导等温扩增(LAMP)技术是近年发展起来的一项新的快速核酸恒温扩增技术,具有高效、快速、特异、易检测、易操作等特点,自面世以来被科学家认为是能替代常规PCR的一项扩增技术,非常适用于现场检测和基层检测。本文就LAMP技术的原理、特点、进展及其在病原微生物检测中的应用进行了简要的概述。     

诱导性多潜能干细胞技术及应用研究进展

 对诱导性多潜能干细胞（iPS细胞）的研究技术进展、临床应用前景以及存在问题等方面进行了综述。     

虚拟显微镜技术及其在医学领域的应用

虚拟显微镜是现代虚拟仪器技术与传统光学显微镜的有机结合,其在光学显微镜有效放大观察的基础上利用程控扫描方式采集高分辨数字成像完成图像拼接制作虚拟切片、搭载计算机和互联网络构建虚拟环境。近年来,虚拟显微镜系统技术在不断进展并且逐步应用于病理学诊断和形态学相关学科的实验教学。     

最新分子细胞生物学方法与技术研究进展

分子细胞生物学是从20世纪60年代迅速发展起来的一门新兴学科,通过对细胞核以及细胞质内的各种超微结构及其功能更为直观的认识,尤其是从分子水平上对其进行深入的探讨,使得分子细胞生物学日臻完善,并且也成为研究生命科学的重要技术.在医学研究领域,体细胞核移植、干细胞定向诱导分化、细胞重编程(特别是诱导性多潜能干细胞技术)的研究为我们制备体外疾病模型、研究疾病发生机制、寻求细胞及组织移植的新材料方面提供了重要的技术支撑.     

浅谈PBL在护理本科生理学教学中运用的一些体会

现代科学技术的进步对教学任务提出了更高的要求。PBL(prob lem based learn ing)适应于这一要求,鼓励学生以问题为中心,在学习过程中探索问题,解决问题。将PBL灵活运用于生理学课堂教学,通过精心设计课堂提问、组织基于问题的专题讨论以及结合实验示教等多种方式,活跃课堂教学气氛、激发学生的探索热情,并充分培养学生的创造性思维能力,从而提高课堂教学质量。     

虚拟病理切片及其应用

病理学是一门形态学,病理切片是病理诊断的主要手段。在疾病尤其是肿瘤诊治过程中．如果出现医疗纠纷,病理切片则是不可缺少的法律依据,同时,病理切片的积累为疾病的流行病调查、病因分析、转归评估以及了解患者不同时期疾病发展与转归等提供可靠的依据。因此,病理切片需要长期保存。目前,病理切片保存存在一些的问题：（1）储存空间有限;（2）切片褪色、损坏;     

奥帕曲拉对内皮素-1诱导的大鼠心肌细胞肥大的影响

 摘要： 目的 探讨奥帕曲拉（OMA）对内皮素-1（ET-1）诱导的大鼠心肌细胞肥大的作用及其机制。方法 经差速贴壁法获得乳鼠心肌细胞,随机分为对照组、ET-1组、OMA 组、ET-1＋OMA组、ET-1＋OMA＋L-NAME组和ET-1＋OMA＋HS-142-1（GC受体阻断剂）组。用分光光度法测定NOS活性, 放射免疫法测定细胞内cGMP水平;液体闪烁计数仪测定3H-亮氨酸（[3H]-leu）掺入率及蛋白激酶C (PKC)活性;用HJ2000图像分析系统测定细胞表面积。结果10-6mol/L ET-1能显著增加心肌细胞表面积、PKC活性及[3H]-leu 掺入率, 降低NOS活性及 cGMP含量 (均P < 0.01); 10-7 mol/L OMA能显著抑制ET-1的上述作用（均P < 0.01）,L-NAME、HS-142-1可部分阻断OMA的作用。结论OMA能抑制ET-1诱导的心肌细胞肥大效应,该作用部分通过NO/cGMP、PKC调控。     

Utility of whole slide imaging and virtual microscopy in prostate pathology

Whole slide imaging (WSI) has been used in conjunction with virtual microscopy (VM) for training or proficiency testing purposes, multicentre research, remote frozen section diagnosis and to seek specialist second opinion in a number of organ systems. The feasibility of using WSI/VM for routine surgical pathology reporting has also been explored. In this review, we discuss the utility and limitations of WSI/VM technology in the histological assessment of specimens from the prostate. Features of WSI/VM that are particularly well suited to assessment of prostate pathology include the ability to examine images at different magnifications as well as to view histology and immunohistochemistry side-by-side on the screen. Use of WSI/VM would also solve the difficulty in obtaining multiple identical copies of small lesions in prostate biopsies for teaching and proficiency testing. It would also permit annotation of the virtual slides, and has been used in a study of inter-observer variation of Gleason grading to facilitate precise identification of the foci on which grading decisions had been based. However, the large number of sections examined from each set of prostate biopsies would greatly increase time required for scanning as well as the size of the digital file, and would also be an issue if digital archiving of prostate biopsies is contemplated. Z-scanning of glass slides, a process that increases scanning time and file size would be required to permit focusing a virtual slide up and down to assess subtle nuclear features such as nucleolar prominence. The common use of large blocks to process prostatectomy specimens would also be an issue, as few currently available scanners can scan such blocks. A major component of proficiency testing of prostate biopsy assessment involves screening of the cores to detect small atypical foci. However, screening virtual slides of wavy fragmented prostate cores using a computer mouse aided by an overview image is very different from screening glass slides using a microscope stage. Hence, it may be more appropriate in this setting to mark the lesional area and focus only on the interpretation component of competency testing. Other issues limiting the use of digital pathology in prostate pathology include the cost of high quality slide scanners for WSI and high resolution monitors for VM as well as the requirement for fast Internet connection as even a subtle delay in presentation of images on the screen may be very disturbing for a pathologist used to the rapid viewing of glass slides under a microscope. However, these problems are likely to be overcome by technological advances in the future.     

Digital pathology: current status and future perspectives

During the last decade pathology has benefited from the rapid progress of image digitizing technology. The improvement in this technology had led to the creation of slide scanners which are able to produce whole slide images (WSI) which can be explored by image viewers in a way comparable to the conventional microscope. The file size of the WSI ranges from a few megabytes to several gigabytes, leading to challenges in the area of image storage and management when they will be used routinely in daily clinical practice. Digital slides are used in pathology for education, diagnostic purposes (clinicopathological meetings, consultations, revisions, slide panels and, increasingly, for upfront clinical diagnostics) and archiving. As an alternative to conventional slides, WSI are generally well accepted, especially in education, where they are available to a large number of students with the full possibilities of annotations without the problem of variation between serial sections. Image processing techniques can also be applied to WSI, providing pathologists with tools assisting in the diagnosis-making process. This paper will highlight the current status of digital pathology applications and its impact on the field of pathology.     

Application of virtual microscopy in clinical cytopathology.

Virtual microscopy (VM) refers to the use of an automated microscope and digital imaging technology to scan, store, and view glass slides. VM systems allow the user to view a scanned image of the entire slide at multiple magnifications on a computer screen. We tested VM to evaluate its possible utility in diagnostic cytopathology. Ten cervical-vaginal monolayered preparations (AutoCyte preparation) were scanned using a BLISS (Bacus Laboratories Inc. Slide Scanner) system. Approximately 20-30% of the cellular area of each slide was imaged. The cases were randomly chosen to include examples ranging from benign cellular changes (BCC) to high-grade squamous intraepithelial lesions (HSIL). The computer performed image tiling and fusing of multiple JPEG images to create a high-quality VM slide. Six examiners (two each of cytopathologists, senior residents, and cytotechnologists) blindly evaluated the VM slides using an image server program (WebSlide Browser thin client software). The cytopathologic diagnoses made on the VM slide were then compared to the original glass slide diagnoses. BLISS took 36-100 min (avg. 58.4 min) to scan the selected fields in a glass slide with file sizes ranging from 23.1-83.6 MB. Time taken by the examiners to render a diagnosis ranged from 1-15 min (avg. 4.1 min) per case. The combined diagnostic accuracy was 98.3%. Only one case of LSIL was missed by one examiner. VM is a promising new tool, which gives a user the feel and simulated experience of an actual microscopic examination and provides a useful alternative to a glass slide in diagnostic cytopathology. Possible applications include: 1) second opinion consultation without transporting the glass slide, 2) education, 3) VM proficiency tests / board exams, and 4) telepathology. Shortcomings include 1) expensive initial setup, 2) inability to maintain an adequate focus in a thick smear with multiple levels, 3) large storage size of the VM slide, and 4) relatively long time needed to scan a slide.     

Automated complete slide digitization: a medium for simultaneous viewing by multiple pathologists.

Developments in telepathology robotic systems have evolved the concept of a 'virtual microscope' handling 'digital slides'. Slide digitization is a method of archiving salient histological features in numerical (digital) form. The value and potential of this have begun to be recognized by several international centres. Automated complete slide digitization has application at all levels of clinical practice and will benefit undergraduate, postgraduate, and continuing education. Unfortunately, as the volume of potential data on a histological slide represents a significant problem in terms of digitization, storage, and subsequent manipulation, the reality of virtual microscopy to date has comprised limited views at inadequate resolution. This paper outlines a system refined in the authors' laboratory, which employs a combination of enhanced hardware, image capture, and processing techniques designed for telepathology. The system is able to scan an entire slide at high magnification and create a library of such slides that may exist on an internet server or be distributed on removable media (such as CD-ROM or DVD). A digital slide allows image data manipulation at a level not possible with conventional light microscopy. Combinations of multiple users, multiple magnifications, annotations, and addition of ancillary textual and visual data are now possible. This demonstrates that with increased sophistication, the applications of telepathology technology need not be confined to second opinion, but can be extended on a wider front.     

Comparing virtual with conventional microscopy for the consensus diagnosis of Barrett’s neoplasia in the AspECT Barrett’s chemoprevention trial pathology audit

Sanders D S A, Grabsch H, Harrison R, Bateman A, Going J, Goldin R, Mapstone N, Novelli M, Walker M M & Jankowski J
(2012) Histopathology  61, 795–800 Comparing virtual with conventional microscopy for the consensus diagnosis of Barrett’s neoplasia in the AspECT Barrett’s chemoprevention trial pathology audit Aims: To compare the diagnostic accuracy of conventional versus virtual microscopy for the diagnosis of Barrett’s neoplasia. Methods and results: Sixty‐one biopsies from 35 ASPirin Esomeprazole ChemopreventionTrial (AspECT) trial patients were given a Barrett’s neoplasia score (1–5) by a panel of five pathologists using conventional microscopy. Thirty‐three biopsies positive for neoplasia were digitized and rescored blindly by virtual microscopy. Diagnostic reliability was compared between conventional and virtual microscopy using Fleiss’ kappa. There was substantial reliability of diagnostic agreement (κ = 0.712) scoring the 61 biopsies and moderate agreement scoring the subgroup of 33 ‘positive’ biopsies with both conventional microscopy (κ = 0.598) and virtual microscopy (κ = 0.436). Inter‐observer diagnostic agreement between two pathologists by virtual microscopy was substantial (κ = 0.76). Comparison of panel consensus neoplasia scores between conventional and virtual microscopy was almost perfect (κ = 0.8769). However, with virtual microscopy there was lowering of the consensus neoplasia score in nine biopsies. Conclusions: Diagnostic agreement with virtual microscopy compares favourably with conventional microscopy in what is recognized to be a challenging area of diagnostic practice. However, this study highlights possible limitations for this method in the primary diagnostic setting.     

Virtual microscopy for learning and assessment in pathology

Virtual slides are high-magnification digital images of tissue sections, stored in a multi-resolution file format. Using appropriate software, these slides can be viewed in a web browser in a manner that closely simulates examination of glass slides with a real microscope. We describe the successful implementation of teaching microscopic pathology with virtual slides and, for the first time, their use in summative assessment. Both students and teaching staff readily adapted to the use of virtual microscopy. Questionnaire feedback from students strongly indicated that virtual slides solved a number of problems in their learning, while providing good to excellent image quality. A deliberate policy of allocating two students per workstation promoted collaboration and helped to maintain interest in microscopic pathology. The use of a secure browser facilitated assessment using virtual slides, with no technical or security issues arising despite high peak demand. The new Medicine programme at the University of New South Wales will exclusively utilize virtual microscopy for the study of both histology and histopathology. We believe that the use of high-quality learning resources such as virtual slides can ensure that microscopic examination of tissues remains both meaningful and interesting.