亚像素断层图像配准数据集的建立

目的针对数字化冰冻铣切断层图像的特点,探讨一种实用的高精度图像配准方法,建立基于数字化断层图像的亚像素级配准数据集。方法采用冰冻铣切技术获取成年男性头颈标本的冰冻连续断层图像,在M atlab软件中自动提取定标点图像特征,采用基于2点的刚体变换算法实现图像的自动配准。结果配准后图像定标点与基准配准点的误差小于1个像素,达到亚像素水平。结论采用外定标的图像配准算法可建立亚像素级的配准数据集,定位标记物的准确识别是获得亚像素级配准数据集的保证。     

A novel method to identify and isolate proliferative inflammatory atrophy (PIA) clusters and to extract high-quality PIA RNA

Epidemiological and histopathological studies have indicated that proliferative inflammatory atrophy (PIA) of the prostate is closely associated with the onset and development of prostate cancer (PCa). However, accurate isolation of PIA still remains a difficult matter, as well as high-quality RNA extraction from isolated PIA. These issues generated a lack of molecular evidence to support the mechanistic explanation proposed for the progression of PIA to PCa. Therefore, the isolation of PIA and the extraction of high-quality RNA from isolated PIA are of great importance to further demonstrate the correlation between PIA and the development of PCa at a molecular level. In this study, clinical samples from radical prostatectomy were stored in liquid nitrogen, PIA was identified by H&E staining of cryosections, PIA clusters were isolated by manual microdissection, total RNA was extracted from the PIA clusters by Trizol, and RNA quality was determined using the Agilent 2100 Bioanalyzer. Our results showed that PIA might be isolated by manual microdissection of cryosections stored in liquid nitrogen from clinical radical prostatectomy and used for extracting high-quality RNA (RIN > 7.5) by Trizol. Therefore, the present study established a valid method to discover molecular evidence in support of the correlation between PIA and the development of PCa.     

Correlative 3D superresolution fluorescence and electron microscopy reveal the relationship of mitochondrial nucleoids to membranes

Microscopic images of specific proteins in their cellular context yield important insights into biological processes and cellular architecture. The advent of superresolution optical microscopy techniques provides the possibility to augment EM with nanometer-resolution fluorescence microscopy to access the precise location of proteins in the context of cellular ultrastructure. Unfortunately, efforts to combine superresolution fluorescence and EM have been stymied by the divergent and incompatible sample preparation protocols of the two methods. Here, we describe a protocol that preserves both the delicate photoactivatable fluorescent protein labels essential for superresolution microscopy and the fine ultrastructural context of EM. This preparation enables direct 3D imaging in 500- to 750-nm sections with interferometric photoactivatable localization microscopy followed by scanning EM images generated by focused ion beam ablation. We use this process to "colorize" detailed EM images of the mitochondrion with the position of labeled proteins. The approach presented here has provided a new level of definition of the in vivo nature of organization of mitochondrial nucleoids, and we expect this straightforward method to be applicable to many other biological questions that can be answered by direct imaging.     

基于冰冻切片的人体骨盆有限元模型的建立与初步验证

 目的 建立基于冰冻切片的人体骨盆有限元模型,以用于研究骨盆非线性生物力学特性。方法 采用“中国数字虚拟人”第1例男性冰冻切片数据,运用切片图像处理软件CryoSegmentation提取出骨轮廓线,分别建立骨皮质、骨松质线模型和有限元模型。骨与软骨选用SOLID92四面体十节点单元,韧带采用2D索单元LINK10,分别赋予材料属性。结果 按骶髂关节为融合状态进行模型计算,应力主要集中部位是椎骨前面、骶髂关节面及周边、髂骨弓、坐骨大切迹、髋臼侧缘以及股骨颈;骶髂关节面应力主要集中在耳状面周缘,最大位移发生在L3;融合状态下各韧带所承拉力都较小。结论 基于冰冻切片的建模方法可获取更为精确的人体解剖结构信息,包含软组织条件的骨盆有限元模型的建立是基于CT和MRI有限元建模方法的重要补充。     

Role of physical factors in the pathogenesis of bullous pemphigoid: Case report series and a comprehensive review of the published work

Bullous pemphigoid (BP) is an autoimmune subepidermal blistering disease affecting mainly the elderly. The subtype of the disease induced by physical agents represents a rare and, therefore, insufficiently characterized form. In the present study, we aimed to contribute to a better understanding of the pathogenetic mechanisms involved in the onset of BP induced by different trigger factors. We have retrospectively analyzed nine cases of BP. All patients were characterized based on clinical, epidemiological and immunological parameters. For each case, the trigger factor involved was specified. In addition to our retrospective analysis, a comprehensive review of the 59 published cases was conducted, regarding the involvement of trigger factor in BP, and clinical, epidemiological and immunological data were collected. In the local study, conducted on nine patients diagnosed with BP, various trigger factors were identified: contrast substance injection, surgical procedure, mechanical trauma, insect bite, thermal burn, radiotherapy and ultraviolet exposure associated with pre‐existing psoriasis. The autoantibodies from all patients were shown to activate granulocytes and induce dermal–epidermal split. Different hypotheses regarding the pathogenetic mechanism involving the trigger factors have been discussed. In regard of the pathogenetic mechanism, we believe that the most reliable hypothesis is that BP patients already have low titers of anti‐basement membrane autoantibodies which activate the granulocytes. However, more studies are needed for a better understanding of the pathogenetic mechanism of the intervention of trigger factors.     

X-Ray microanalysis of the secretory granules in goblet cells of mouse intestinal tracts: Changes with age

Changes in the elements in goblet cell secretory granules in three portions of the intestinal tract — duodenum, proximal colon, and distal colon — in mice from 14 days to 18 months after birth were studied by quantitative electron probe X-ray microanalysis on quick-frozen and freeze-dried cryosections by calculating the ratio of each element. In the analysis of five elements, the peaks decreased in the order of sulfur (S), potassium (K), chlorine (Cl), phosphorus (P), and calcium (Ca) in the duodenum at 1 month; in the proximal colon, the peaks declined in the order of K, S, Cl, Ca, and P at 3 months; and in the distal colon, the peaks declined as S, K, Ca, Cl, and P at 3 months. The highest average ratio of S was obtained in the duodenum at 1 month, and this value then declined with age. In the proximal colon, the average ratio of K was the highest at 3 months, then declined afterward with age. The highest average ratio of S was at 3 months, and the highest average ratio of K was at 1 month in the distal colon. The ratio of peak counts to the background was calculated for each type of granule. Significant differences were found in the ratios of S and K between the proximal and distal colon. As for S, the duodenum and the distal colon had the highest ratios and the proximal colon had the second highest. Concerning K, the proximal colon had the highest and the duodenum and the distal colon the second highest ratio. The secretory granules of goblet cells in the three portions of the intestinal tract were shown to contain different amounts of each element. Especially, the amount of S differed most from the proximal to the distal colon.This paper was presented in part at the 25th Annual Meeting of the Clinical Electron Microscopy Society of Japan in Matsumoto, September 1993; the 27th Annual Meeting of the Clinical Electron Microscopy Society of Japan in Kurashiki, September 1995; and the 28th Annual Meeting of the Clinical Electron Microscopy Society of Japan in Osaka, September 1996.     

X-ray microanalysis of the secretory granules in the intestinal goblet cells of aging mice

The elemental changes in the goblet cell secretory granules of the mouse jejunum and ascending colon during postnatal development and aging were studied by a quantitative electron probe X-ray microanalysis on quick frozen and freeze-dried cryosections. In the jejunum, sulfur showed the highest peaks, followed by those for K, Cl, Ca, P and Na, respectively. In the colon, on the other hand, potassium showed the highest peaks, followed by those for S, Cl, Ca, P, Na respectively. Sulfur in the jejunum was at its lowest on postnatal day 14, as its highest at 1 month, and then gradually decreased with aging.     

3D Cryo‐Imaging: A Very High‐Resolution View of the Whole Mouse

We developed the Case Cryo‐imaging system that provides information rich, very high‐resolution, color brightfield, and molecular fluorescence images of a whole mouse using a section‐and‐image block‐face imaging technology. The system consists of a mouse‐sized, motorized cryo‐microtome with special features for imaging, a modified, brightfield/fluorescence microscope, and a robotic xyz imaging system positioner, all of which is fully automated by a control system. Using the robotic system, we acquired microscopic tiled images at a pixel size of 15.6 μm over the block face of a whole mouse sectioned at 40 μm, with a total data volume of 55 GB. Viewing 2D images at multiple resolutions, we identified small structures such as cardiac vessels, muscle layers, villi of the small intestine, the optic nerve, and layers of the eye. Cryo‐imaging was also suitable for imaging embryo mutants in 3D. A mouse, in which enhanced green fluorescent protein was expressed under gamma actin promoter in smooth muscle cells, gave clear 3D views of smooth muscle in the urogenital and gastrointestinal tracts. With cryo‐imaging, we could obtain 3D vasculature down to 10 μm, over very large regions of mouse brain. Software is fully automated with fully programmable imaging/sectioning protocols, email notifications, and automatic volume visualization. With a unique combination of field‐of‐view, depth of field, contrast, and resolution, the Case Cryo‐imaging system fills the gap between whole animal in vivo imaging and histology. histology. Anat Rec, 292:342–351, 2009. © 2009 Wiley‐Liss, Inc.     

适于小动物研究的断层解剖成像系统

目的研制一种具有高空间分辨率、适于实验小动物研究的断层解剖成像系统。方法利用冰冻铣削的方式逐层去除一定厚度的实验小动物冷冻标本,获得实验小动物断层解剖数据集。断层切削设备的最小进给量为0.001mm,重复定位精度为0.01／300mm。本文以1只成年SD大鼠为例,叙述了断层解剖成像的实验过程,切削间距为0.02mm。结果展示了部分断层解剖图像及三维重建结果。获得大鼠数据集,体素尺寸为0.02mm×0.02mm×0.02mm,9475幅,314.68 Gb,图像分辨率4600×2580×24bit。结论该系统能够完整地获得小动物精细结构信息,为小动物研究提供了一种新的有效手段。     

Insulin-like growth factor 1 (IGF-1) receptors in the human brain: quantitative autoradiographic localization

The distribution of Insulin-like growth factor 1 (IGF-1) receptors in large cryosections of human brain hemispheres (80-microns) was studied by quantitative autoradiography using 125I-IGF-1 as ligand. Postmortem tissue only from individuals free from neurological diseases was used. The highest densities of IGF-1 receptors were found in the hippocampus, amygdala and parahippocampal gyrus. Intermediate densities were observed in the cerebellum, cerebral cortex and caudate nucleus, whereas low densities of IGF-1 receptors were obtained in the substantia nigra, red nucleus, white matter and cerebral pedunculus. The cartography of IGF-1 receptors in the normal human brain will hopefully be of use in the study of the alteration of these receptors in diseased brain.