OBJECTIVEIt is well known that glioma stem cells-progenitors (GSCP) proliferate indefinitely and hardly differentiate in vitro, however, the reasons remain unknown. The aim of this study was to explore the ultrastructural basis of GSCP.

METHODSGSCP, kept by our laboratory, were collected, embedded, and cut into ultrathin sections and observed under the transmission electron microscope.

RESULTSA single GSCP usually had relatively well developed mitochondria, Golgi apparatuses, ribosomes, and undeveloped rough endoplasmic reticulum, but seldom lysosomes and no typical autophagosomes were found, and the nuclear-cytoplasmic ratio was high. The nuclei frequently contained huge amounts of euchromatin and a small quantity of heterochromatin, and in most nuclei there were only one nucleolus, however, two or more nucleoli were also common. Typical apoptotic cells could hardly be found in tumor-spheres, and between neighboring cells in tumor-spheres there were incompletely developed desmosomes or intermediate junction.

CONCLUSIONThe ultrastructural features of glioma stem cells-progenitors showed that BTSCP were very primitive and the lack of autophagy and the underdevelopment of some other cellular organelles are probably the reasons for the differential inhibition of GSCPs.

Brain Neoplasms ; ultrastructure ; Cell Line, Tumor ; Cell Membrane ; ultrastructure ; Cell Nucleus ; ultrastructure ; Chromatin ; ultrastructure ; Cytoplasm ; ultrastructure ; Glioma ; ultrastructure ; Humans ; Intercellular Junctions ; ultrastructure ; Microscopy, Electron, Transmission ; Mitochondria ; ultrastructure ; Neoplastic Stem Cells ; ultrastructure

A hybrid segmentation algorithm is proposed for automatic segmentation of blood cell images based on adaptive multi-scale thresholding and seeded region growing techniques. Firstly, an adaptive and scale space filter (ASSF) is applied to image histogram and a scale space image is built. According to the properties of the scale space image, proper thresholds can be obtained to separate the nucleus from the original image and the white blood cells are located. Secondly, the local color similarity and global morphological criteria constrain seeded region growing in order to finish the segmentation of the cytoplasm. The detection accuracy of white blood cell is 98% and the segmentation accuracy based on the subjective evaluation is 93%. Test shows that this algorithm is effective for automatic segmentation of white blood cells.
Algorithms ; Automation ; Blood Cells ; Cell Nucleus ; ultrastructure ; Color ; Cytoplasm ; ultrastructure ; Humans ; Image Enhancement ; Leukocytes

The effects of morphine HCI on the rat mesenteric mast cells were studied with the electron microscopy. The materials were prepared for electron microscopy by osmium tetroxide fixation and embedding in Epon. The rat mesenteric mast cells showed no distinct morphological changes due to morphine HCl, but the mast cell granlues were changed in various ways. For instance, they formed dusters, showed granular lysis, and an appearance of electron transparency. Frequently, some granules appeared in the extracellular space and the boundary of the granules was not evident. From the results mentioned above, it was suggested that rat mesenteric mast cell granules were affected by morphine HCl in the shape, the granular matrix, and the granular boundaries.
Animal ; Cell Nucleus/ultrastructure ; Cytoplasm/ultrastructure ; Cytoplasmic Granules/drug effects ; Cytoplasmic Granules/ultrastructure ; Golgi Apparatus/ultrastructure ; Male ; Mast Cells/drug effects ; Mast Cells/ultrastructure* ; Mesentery/drug effects ; Mesentery/ultrastructure* ; Microscopy, Electron ; Mitochondria, Muscle/ultrastructure ; Morphine/pharmacology* ; Rats

INTRODUCTIONAn investigation was carried out to determine the morphological characteristics of fibroblasts in two portions of the vocal fold (VF) mucosa, the macula flava (MF) and Reinke's space (RS), of three different age groups: newborns, adults and geriatrics.

METHODSNormal human VF obtained from autopsy cases were included in this study: four from mature newborns; four from middle-aged adults; and four from geriatric cases. Fibroblasts in RS and MF were investigated by transmission electron microscopy.

RESULTSThe fibroblasts of the MF in both adults and newborns tended to be stellate in shape, with a small nucleus/cytoplasm (N/C) ratio and a well-developed rough endoplasmic reticulum (rER) and Golgi apparatus (GA). Most of the fibroblasts present in RS were oval in newborns and spindle-shaped in adults, with a large N/C ratio and less developed rER and GA. The majority of fibroblasts of the geriatric MF were stellate in shape; while in geriatric RS, the majority of fibroblasts were spindle-shaped with an N/C ratio of 0.5 to 2.0 as in the case of adults. However, the development of rER and GA was less marked in geriatrics than in adults.

CONCLUSIONHistological changes of fibroblasts in the VF mucosa are one of the important causes of the change in voice quality with ageing. Furthermore, geriatric changes in the vocal ligament can be attributed to the activities and the presence of ageing processes in fibroblasts of geriatric VF mucosa.

Adult ; Age Factors ; Aged ; Cell Nucleus ; ultrastructure ; Cytoplasm ; ultrastructure ; Endoplasmic Reticulum ; ultrastructure ; Female ; Fibroblasts ; ultrastructure ; Golgi Apparatus ; ultrastructure ; Humans ; Infant, Newborn ; Laryngeal Mucosa ; ultrastructure ; Male ; Microscopy, Electron, Transmission ; methods ; Vocal Cords ; ultrastructure

The Spindle-view, a specialized instrument for observing spindle image, was applied to observe the meiotic spindles of vitro matured porcine oocytes at 36, 42, 44, 48h, and enucleation from porcine, comparing to the previously methods (McGrath-Solter's method and two-step-squeezing method) in the enucleated. The results showed that: (1) there was no noticeable differences at vicinity of spindle images and 1st polar body among in vitro matured porcine oocytes at 40-48 h under the instrument; (2) Spindle-view is suitable for the observation of meiotic spindles of matured oocytes and enucleation from porcine; the modified Spindle-view method for enucleation is significantly better than McGrath-Solter' s method and two-step-squeezing method in the enucleated rates (95.5%, 42.1%, 74.2%, P < 0.0l) of absolutely removing nuclei matter; (3) the spindle images could be used to monitor the oocyte qualities.
Animals ; Cell Nucleus ; ultrastructure ; Cells, Cultured ; Cytological Techniques ; Female ; Nuclear Transfer Techniques ; Oocytes ; cytology ; Spindle Apparatus ; ultrastructure ; Swine

Promyelocytic leukemia protein (PML) is a major component of PML nuclear bodies (PML NBs). Fusion of promyelocytic leukemia gene (PML) with retinoic acid receptor alpha gene with the t (15;17) translocation causes disassembly of PML NBs, leading to development of acute promyelocytic leukemia. In contrast, PML overexpression as well as different morphological changes of PML NBs were described in a few solid tumors. In this study, the expression of PML through the multistep hepatocarcinogenesis was analyzed in 95 cases of human hepatocellular carcinomas (HCCs) for comparison along with dysplastic nodules (DNs) and background liver cirrhosis (LC) or chronic hepatitis by immunohistochemistry and immunoblot. In addition, cases of HCCs were further evaluated according to their histologic grade and etiology. The amount of PML as well as the num-ber and size of PML NBs increased gradually through the progression from LC, DNs to HCCs. The overexpression of PML in HCCs was much more closely associated with HBV infection than HCV infection or alcoholic liver disease. The PML expression, however, was not correlated with histologic grade of HCCs. These results suggest that PML is involved in the early stage of multistep hepatocarcinogenesis, and HBV infection may be associated with the overexpression of PML and the morphological alteration of PML NBs.
Carcinoma, Hepatocellular/*chemistry/ultrastructure ; Cell Nucleus/*chemistry ; Human ; Liver/chemistry ; Liver Neoplasms/*chemistry/ultrastructure ; Neoplasm Proteins/*analysis ; Precancerous Conditions/*chemistry/ultrastructure ; Transcription Factors/*analysis ; Tumor Cells, Cultured

To investigate toxicity of aflatoxin Gl and its mechanism, light microscopic, histochemical, electron microscopic and autoradiographic studies were done on the rat liver at various time intervals after the administration of aflatoxin Gl. Light microscopic alteration was first observed at 6 hours and necrosis of periportal hepatic cells was found at 18 hours. However, reduction of Feulgen positivity of the nucleus and pyroninophilia of cytoplasm was observed as early as 1 hour. Ultrastructural changes were noted at 6 hours and were advanced at l8 hours. Early changes consisted of nucleolar segregation, dilatation of rough endoplasmic reticulum, swelling of mitochondria and detachment of membrane bound ribosomes followed later by disruption of cytoplasmic organellae and focal necrosis. These changes were most marked at periportal region. Autoradiographic studies showed inhibition of H3-uridine incorporation into the nucleus at 1 hour, was most marked at 6 hours, and showed some recovery at 18 hours. H3-uridine labeling in the cytoplasm was also inhibited and the most marked inhibition was noted at 1 hour after the aflatoxin administration. These data indicate aflatoxin Gi has a hepatotoxic effect, particulary at the periportal region. This toxic effect is likely due to inhibition of nuclear RNA synthesis which leads to inhibition of ribosomal RNA and eventually protein synthesis. The DNA synthesis is also inhibited, as shown by reduction of Feulgen reaction in the nucleus.
Aflatoxins/toxicity* ; Animal ; Autoradiography ; Cell Nucleus/ultrastructure ; Cytoplasm/ultrastructure ; Histocytochemistry ; Liver/ultrastructure* ; Microscopy, Electron ; Mitochondria, Liver/drug effects ; Rats ; Uridine/metabolism

A simple method of trypsin/collagenase I alternative digestion and iterative culture flask adherence to discard fibroblasts for bovine mammary cell culture was established in this study. By immunohistochemistry, flow cytometry, western blot, Electron microscopy analysis, the characteristics of bovine mammary cells were investigated in vitro. Effect of hyperthermia on the cell ultrastructures was also observed. The results showed that the mammary cells were diploid epithelia with intact 30 pairs chromatins, which could secrete alpha-casein into the medium. After exposed to hyperthermia, the cell condensed chromatin like crescent on the nuclei verges, mitochondria occurred expansion and vacuolization, and apoptotic bodies appeared, which suggested that heat stress could induce apoptosis of the mammary epithelia.
Animals ; Apoptosis ; Blotting, Western ; Caseins ; metabolism ; Cattle ; Cell Line ; Cell Nucleus ; ultrastructure ; Cells, Cultured ; Chromatin ; ultrastructure ; Epithelial Cells ; cytology ; metabolism ; ultrastructure ; Female ; Flow Cytometry ; Hot Temperature ; Immunohistochemistry ; Mammary Glands, Animal ; cytology ; metabolism ; Microscopy, Electron, Transmission ; Mitochondria ; ultrastructure ; Vimentin ; metabolism

The effect of four alkylating agents (MMS, EMs, DMN, DEM), under various con centrations on mouse bone marrow erythrocytes, were studied by means of the micronucleus test. The results obtained were as follows: 1) The lethal doses on mice were MMS = 130 mg/kg/bw, EMS = 300 mg/kg/bw, DMN = 50 mg/kg/bw and DEN = 70 mg/kg/bw. 2) Micronuclei were easily seen and in different controls the micronulei were found a little over 0.1%. 3) The dose-effect relationship was obtained. In the MMS and EMS treated groups, incidences of micronulei were 0.45 to 2.56% and 0.4 to 2.1% respectively. 4) In the DMN and DEN treated groups, incidences varied between 0.15 to 0.90 % and 0.2 to 1.02% respectively. 5) Four alkylating agents were compared and discussed with respect to micro nucleus production from chromosomal aberrations.
Animal ; Bone Marrow/ultrastructure ; Carcinogens/pharmacology* ; Cell Nucleus/drug effects* ; Chromosome Aberrations* ; Erythrocytes/ultrastructure ; Female ; Mesylates/pharmacology* ; Mice ; Mutagens/pharmacology ; Nitrosamines/pharmacology*

Cell apoptosis induced by duck reovirus (DRV)in duck embryo fibroblasts (DEF) was ascertained by light microscope and electron microscopy, DNA Ladder, flow cytometry and fluorescent microscopy. Typical morphological apoptotic features including cell shrinkage and condensation, margination of nuclear chromatin were observed under light microscope and the formation of apoptotic bodies by electron microscopy. DNA ladder was shown by DNA fragment analysis at 24-144h post infection. Flow cytometry showed that the cell apoptosis appeared at 24h and reached it's crest-time at 72-96h, decreased at 144h. Fluorescent microscopy showed that the apoptotic cells which showed green fluorescence appeared at 24h, the number of dead cells which showed red fluorescence increased with the time went by. The results above confirmed that the apoptosis of DEF was successfully induced by DRV.
Animals ; Apoptosis ; physiology ; Cell Nucleus ; ultrastructure ; Cells, Cultured ; DNA Fragmentation ; Ducks ; Embryo, Nonmammalian ; cytology ; Fibroblasts ; cytology ; ultrastructure ; virology ; Flow Cytometry ; Host-Pathogen Interactions ; Microscopy, Electron, Transmission ; Reoviridae ; physiology