大鼠肝细胞线粒体三维重建和体视学分析

为深入研究大鼠肝细胞线粒体的形态特征，本文以连续超薄切片追踪观察不同形态线粒体的三维结构，重建其三维模型，把线粒体划分为简单型、复杂型、特殊型三个形态类型，并对不同类型的线粒体进行体视学分析。结果显示：简单型线粒线（球形、盘球及杆形等）占线粒体总数的５３９％，其体密度为８９６％；复杂型线粒体（分枝型及各种不规则形等）占４２０１％，其体密度为８６７％；而特殊型线粒体（芽生状等）占４６０％，其体密度仅为０９７％。这表明在正常的肝细胞中复杂型线粒体的含量并不为少，线粒体形态的多样性与其自身的发育相对应。分析还显示不同类型线粒体嵴膜密度和基质颗粒数密度有所差异，表明线粒体嵴的生长并不与线粒体体积的增长同步增加，因此，三种类型线粒体的功能状态亦可能有所不同。     

缺碘大鼠甲状腺上皮细胞超微结构观察及其体视学研究

本文动态观察缺碘大鼠动物模型甲状腺上皮细胞超微结构的变化, 同时应用现代体视学技术量化形态学指标, 进行统计学分析。电镜下可见缺碘大鼠甲状腺上皮细胞线粒体、内质网扩张, 随着缺碘时间的延长线粒体进一步肿胀, 出现嵴断裂、缺失、空泡变性, 内质网扩张、膜断裂、脱颗粒。体视学测量结果显示, 线粒体的珡V 及-S均显著高于对照组, 而Nv 和S/V 均显著降低, 表明缺碘使线粒体体积增大, 被高度扩张的内质网包围的线粒体数量减少, 其数密度(Nv) 显著降低, 因此线粒体虽有扩张, 但其Vv 、Sv 仍相对降低。甲状腺上皮细胞中内质网珡V、-S、Vv 和Sv 显著高于对照组、S/V 显著降低, 表明缺碘使内质网高度扩张、体积增大; 扩张的内质网形状不规则, 致使其-S明显增大; 单位体积内内质网的数量减少, Nv 明显降低。本文结果提示, 形态学与体视学相结合为组织形态学研究提供了有效可行的方法学基础     

连续切片的计算机三维重建及立体视觉

本文利用计算机三维重建技术重建颞骨连续切片,恢复其内结构如迷路、面神经、鼓索神经、听骨链、肌腱等的原有三维形态,通过调色、移去部分结构、旋转使结构间关系更清晰。应用立体镜及立体图对方法产生重建结构的立体影像,强化了对所研究结构的三维形态及其空间关系的理解。结果证明此方法是形态学、体视学、计算机科学综合所形成的一个新的有力的解剖工具。     

乙醇对胎鼠肝细胞超微结构的影响及体视学分析

BALB/c妊娠小鼠用20％乙醇作为饮水,平均每d每kg体重摄入乙醇15g。与对照组相比,乙醇组胎鼠肝细胞胞质溶解,部分线粒体肿胀,嵴排列紊乱、溶解破坏,外膜破裂。部分线粒体小、基质电子密度高。有些粗面内质网池扩张、脱颗粒。胞质溶解区域见较多脂肪空泡和溶酶体。毛细胆管普遍扩张,激绒毛短小。经体视学测量,乙醇组线粒体、粗面内质网体积密度与对照组相比有显著下降,具统计学意义。线粒体截面数密度和数密度均下降。结果表明乙醇能导致胎鼠肝细胞超微结构的损害。     

人体乳腺单纯癌癌细胞超微结构的定量研究

应用体视学方法对１０例晚期女性乳腺单纯癌癌细胞线粒体、溶酶体、粗面内质网、核糖体的２８个形态结构参数进行体视学测定和分析。结果发现，癌细胞与正常细胞比较，４种细胞器的有关体积、表面积、面数密度及数密度等１７种形态参数有高度显著和显著差异；线粒体、粗面内质网形态结构均发生了显著变化，核糖体的数目明显增多。上述研究为今后研究不同期乳腺癌癌细胞超微结构的定量变化打下基础，并力求对乳腺癌的分期及病因提供较准确的定量依据。     

大鼠肾小囊上皮细胞发生发育的变化

目的探讨胚胎及生后大鼠肾小囊上皮细胞发生发育的变化规律。方法采用光镜连续切片技术,电镜制作技术,并结合体视学分析方法,对不同发育阶段大鼠肾小囊上皮细胞发育的变化进行形态学观察和体视学测量。结果上皮细胞在发育过程中,细胞排列从紧密到分散,胞质从少到多,胞体从柱状到扁平,细胞突起从无到有,从少到多。结论随着发育过程中形态的变化,上皮细胞逐渐参与了肾小体滤过屏障的组成。     

肝细胞癌癌组织三维形态学观察——共聚焦激光扫描显微镜的应用

目的 应用CLSM对肝细胞癌标本的石蜡切片进行三维重建,显示立体形态特点,探讨CLSM在肿瘤病理学中的应用.方法 30μm石蜡切片,YOYO-1碘化物DNA荧光探针标记细胞核,CLSM连续光学切片、三维重建,观察肝细胞癌、不典型增生及正常肝组织的三维形态特点.结果 肝癌细胞核排列紊乱、积堆,体积增大、大小不等、明显多形;核表面不规则、凹凸不平、荧光强度不均;核内染色质分布不均、结构粗糙、多浓集于核膜下.结论 CLSM的断层扫描和三维重建功能提供了高精密度的三维图象和丰富信息,有助于准确诊断.     

肝细胞癌组织三维形态学观察——共聚焦激光扫描显微镜的应用

目的 应用ＣＬＳＭ对肝细胞癌标本的石蜡切片进行三维重建，显示立体形态特点，探讨ＣＬＳＭ在肿瘤病理学中的应用。方法３０μｍ石蜡切片，ＹＯＹＯ－１碘化物ＤＮＡ荧光探针标记细胞核，ＣＬＳＭ连续光学切片、三维重建，观察肝细胞癌、不典型增生及正常三维形态特点。结果 肝癌细胞排列紊乱、积堆、体积增大、大小不等，明显多形；核表面不规则、凹凸不平、荧光强度不无关 ，核内染色质分布不均、结构粗糙、多浓集于核膜下。结     

鼠实验性肝癌超微结构和ras p21免疫电镜观察

目的：动态观察二乙基亚硝胺诱发大鼠肝细胞肝癌过程超微结构和ｒａｓｐ２１免疫电镜改变。方法：对６０只二乙基亚硝胺诱癌的大鼠分期处死,进行电镜及ｒａｓｐ２１免疫电镜观察。结果：早期即见肝细胞内线粒体、粗面内质网、滑面内质网和核、核仁等显示一系列损伤性变化,小肝细胞灶性增生,并向嗜碱细胞和异常肝细胞转化,到诱癌中期以至诱癌晚期,异型肝细胞逐渐向癌细胞转化,并逐渐形成了癌细胞的超微结构特点。ｒａｓｐ２１免疫电镜观察发现,ｐ２１阳性颗粒除沿细胞膜分布外,还可见到内质网上有免疫反应物沉积,甚至在核膜上也观察到略弱的ｐ２１免疫复合物沉积。结论：随诱癌时间的延长,肝细胞逐渐向癌细胞转化,ｒａｓｐ２１阳性颗粒可在癌变细胞内多处分布。     

三维结构与体视学

基于2012--2013体视学学科发展报告项目和中国体视学学会组织的集体研讨,本文回顾了在体视学（stereology）发展过程中其定义的多样化,探讨了体视学的学科基本特征,进而对体视学的定义与内涵提出新的建议。体视学是不断发展中的研究三维结构的交叉学科和方法学,同时包括基于低维截面或投影数据对三维结构表征参量的统计性推估与结构的三维重建。从投影数据重建三维CT图像属于体视学研究范畴;针对维数大于3的高维结构（三维几何结构＋时间维或其他维）开展体视学理论与方法研究亦很有必要。目前体视学已形成了坚实的理论与方法体系,同时又进入了一个不同方法相互补充、借鉴和协同融合发展的新阶段,有着光明的发展和应用前景。不断推广应用体视学与研究发展体视学新方法仍然是我们面临的两大主题。     

Changes in the relative distribution and properties of smooth endoplasmic reticulum subfractions isolated from the livers of rats fed 2-acetylaminofluorene

A marked increase in the amount and relative proportion of fraction II of the smooth endoplasmic reticulum (SER II) and a concomitant decrease in the SER I fraction were observed in the liver of male Sprague-Dawley rats fed diets containing 0.05% (w/w) 2-acetylaminofluorene (AAF) for various lengths of time (3-18 weeks). The amount of rough endoplasmic reticulum (RER), which remained at control levels after 3 weeks of AAF feeding, i.e. before the appearance of hyperplastic nodules (HPN), was clearly decreased after 18 weeks of continuous AAF-feeding (HPN present). When the rats were subjected to 4 or 5 cycles of interrupted AAF feeding, similar increases in SER II were also observed both in the homogenates of HPN-containing livers, as well as in the homogenates of HPN dissected out from the surrounding liver tissue. SER II, the predominant microsomal membrane fraction in the livers of rats fed AAF for 3 weeks, showed the highest level of induction of epoxide hydrolase. A marked elevation of the manganese-dependent enhancement of polysome binding in vitro was also observed in SER II from the livers of AAF-fed rats.     

Gonadotropin-induced proliferation of endoplasmic reticulum in an androgenic tumor and its relation to elevated plasma testosterone levels.

The development of gonadotropin-induced structural change in a testosterone-secreting Leydig cell tumor and its relationship to plasma testosterone levels were studied in castrate, tumor-bearing mice 2 hr after daily injections of human chorionic gonadotropin (HCG). Tumor cells from control animals were small, averaging less than 1.0 X 10(-9) ml in volume, and were poorly differentiated, having very little smooth endoplasmic reticulum (SER) in their cytoplasm. Average plasma testosterone levels in these mice were near 1.3 ng/ml. Although a five-fold rise in plasma testosterone was measured 2 hr after the first HCG injection, no changes in the endoplasmic reticulum were detected at this time. After the second injection, plasma testosterone rose to only twice control levels, while ultra-areas of mixed rough endoplasmic reticulum and SER. Modest amounts of SER had appeared in many tumor cells after the third injection, and plasma testosterone showed a six-fold rise. Between the third and fourth injections, cell volume increased by about 70% as large accumulations of SER appeared in the cytoplasm of most cells. Plasma testosterone again rose to five times control levels. Increased cell volume and abundant SER were maintained by continuing daily injections of HCG, while the 2-hr plasma testosterone response persisted at five to six times the control level. These findings show that a maximal elevation of plasma testosterone can occur prior to changes in the endoplasmic reticulum of Leydig tumor cells. However, they also suggest that daily repetition of the maximal functional response requires that the cells acquire large quantities of newly produced SER.     

Short-term induction of neoplastic nodules in the rat liver. II. Study of their development and the effects of withdrawal of 2-acetylaminofluorene

Neoplastic liver nodules were induced by a single administrationof N-nitrosodiethylamine (NDEA) and selectively stimulated togrowth by 2-acetylamino-fluorene (2-AAF) and partial hepatectomyto provide morphological data on very early and late stagesof pre-neoplastic development. Presumptive preneoplastic livercells were recognizable by light and electron microscopy by2 days after partial hepatectomy, and they developed within2 weeks into large, solid nodules consisting of plates of 2or 3 cell layers thick, that compressed the surrounding non-nodulartissue. The cells showed nuclei with an enlarged nucleolus,tortuously dispersed rough endoplasmic reticulum (RER) and anelectron lucent cytoplasm. These features remained present throughoutthe first 2 weeks of growth. During this period the initiallysmall, distinct smooth endoplasmic reticulum areas proliferatedgradually. They were associated with an increasing incidenceof cytoplasmic membranes whorl formation and incidently withannulate lamellae. After withdrawal of 2-AAF the majority ofthe nodules regressed resulting in slowly disappearing fociof glycogen rich cells. A relatively small number of solid nodulespersisted. They were characterized by large cells with a homogeneouseosinophilic cytoplasm, enlarged nucleoli and dispersed RER.Because these characteristics were absent in nodular cells thathad reverted to normal liver cells, it is not appropriate toconsider these characteristics to represent neoplastic transformationper se.     

Membrane lipids of hepatic tissue. II. Phospholipids from subcellular fractions of liver and hepatoma 7288CTC

Phospholipids from homogenate, nuclei, mitochondria, endoplasmic reticulum (ER), plasma membrane (PM), and cytosol of liver and hepatoma 7288CTC (from inbred male BUF rats) were analyzed for their concentrations, fatty acid compositions of individual lipid classes, and levels of octadecenoate positional isomers. The phospholipid concentrations of hepatoma mitochondria and ER were less than 60% of liver values. Sphingomyelin concentrations were elevated dramatically in hepatoma nuclei, mitochondria, and PM. Hepatoma nuclei and mitochondria contained only 25% or less the concentrations of phosphatidylethanolamine (PE) as those of liver, whereas ER, PM, and cytosol fractions of hepatoma contained equal or greater concentration of PE than did the corresponding liver fractions. The fatty acid profiles of the individual lipid classes were somewhat characteristic of liver organelles but not of hepatoma. Lipid classes thought to be located preferentially on the outer bilayer of liver PM contained lower percentages of polyunsaturated fatty acids than did hepatoma PM. For hepatoma generally the lipid classes tended to exhibit a more uniform fatty acid profile among organelles, the polyunsaturated fatty acid percentages were decreased, and the octadecenoate percentages were increased. Octadecenoates isolated from individual lipid classes of organelles contained high levels of cis-vaccenate, in addition to oleate, and some class and organelle specificity was observed in liver. In contrast, hepatoma octadecenoates exhibited little class or organelle specificity, and much higher oleate concentrations were found in PM phosphatidylcholine, PM PE, and ER PE in hepatomas than in liver.     

Ultrastructure of primary clear cell carcinoma of the liver

Surgical specimens of 175 cases of primary hepatocellular carcinoma were prepared by routine paraffin section and HE stain. The clear cell cancer specimens were stained with PAS. All the specimens were observed by light microscope. Ultrathin sections were made for 50 samples and studied by electron microscope. Under the light microscope, 79 (45.1%) showed varying amounts of clear cells. According to the proportion and distribution of these cells, clear cell carcinoma of the liver was divided into three types: scattered type (16 cases, 20.3%), localized (43, 54.4%) and diffuse types (20, 25.3%). The clear cancer cells could be found in hepatoma with various degrees of differentiation. Of these 79 cases, 4 (5.1%) were grade I, 53 (67%) and 22 (27.9%) were grades II and III. Positive PAS stain gives an evidence of glycogen in the clear cell cytoplasm. 7 diffuse type clear cell hepatomas were observed with electron microscope. The cytoplasm had only fewer organelles, leading to a void appearance. The amount of the rough endoplasmic reticulum, free ribosome and polyribosome was markedly decreased. So was mitochondria, usually showing swelling and abnormality. The residual rough endoplasmic reticulum and mitochondria were out of normal arrangement. They often aggregated on one side of the nucleus or near the cell membrane. Glycogen particles were increased in some cell cytoplasms. Some particles were even and fine, some were aggregated into masses or scattered. Nuclei showed abnormalities mild to moderate. The nature of the clear cells in liver cancer is the variance of glycogen or lipid in the cytoplasm.(ABSTRACT TRUNCATED AT 250 WORDS)     

放射性碘致大鼠甲状腺损伤超微结构的形态计量学研究

本文应用透射电镜观察了在较低剂量１３１Ｉ作用大鼠甲状腺滤泡上皮细胞超微结构的变化特点，分析了线粒体的变异发生率，并应四川大学研制的ＭＩＡＳ－３００图像分析系统和用形态计量学方法，测定了胞质内粗面内质网的二维和三维形态计量学参数。结果表明：所测的各项参数都有随剂量增高而增大的趋势，并呈现较好的效应剂量依赖关系。由此计算的Ｄ２５值和Ｄ５０值，以粗面内质网体积密度的变化最为明显。提示，低剂量１３１Ｉ作用于滤泡上皮细胞，其早期应重视内质网的损伤效应，以及由此导致合成类固醇激素和外源性蛋白质的障碍     

Particle--lamella complexes in a case of human benign prostate hyperplasia: brief communication.

Particle--lamella complexes (PLC's), described for the first time, were found in glandular epithelial cells of the hyperplastic prostate tissues from a patient with transitional cell carcinoma of the urinary bladder. PLC's observed in this patient were similar to those seen in human hematopoietic neoplastic cells. They showed cylindroid forms and were composed of concentrically arranged lamellae and particles found in rows between these lamellae. PLC is closely related to rough endoplasmic reticulum (RER), and some PLC's were completely surrounded by RER. Particles approximately 25--30 nm in diameter were similar to ribosomes in size, shape, and electron density; lamellae approximately 10 nm thick appeared circular in cross sections and lamellar in longitudinal sections. Although the nature and function of PLC's are as yet unknown, the present observation indicated that PLC's are not a characteristic structure restricted to malignant tumors of hematopoietic origin.