To demonstrate the effect of AB serum on terminal erythroid differentiation ex vivo.
 Methods: After separation of CD34+ cells from cord blood, the cells were cultured and divided into a control group and an experimental group. The effects of AB serum were examined by the expressions of different markers (GPA, Band3 and α4-integrin) for erythroblast differentiation and enucleation by flow cytometry. 
 Results: The CD34+ cells were successfully differentiated to enucleated red blood cells. There were evident differences among the expressions of GPA, Band3 and α4-integrin between the 2 groups. The percentage of GPA positive cells in the experimental group was bigger than that in the control group in every time point. The expression of Band3 in the experimental group was higher than that in the control group. The expression of α4-integrin in the experimental group was lower than that in the control group. In addition, the enucleation rate in the experimental group was higher than that in the control group.
 Conclusion: AB serum can promote the cell differentiation and enucleation during terminal erythroid differentiation in vitro.
ABO Blood-Group System ; blood ; physiology ; Anion Exchange Protein 1, Erythrocyte ; metabolism ; Antigens, CD34 ; blood ; Cell Differentiation ; genetics ; physiology ; Cell Nucleus ; Cells, Cultured ; Erythrocytes ; physiology ; ultrastructure ; Erythropoiesis ; genetics ; physiology ; Fetal Blood ; cytology ; physiology ; Flow Cytometry ; Glycophorins ; metabolism ; Humans ; Integrin alpha4beta1 ; metabolism

To investigate the morphogenetic process of human adenovirus type 41 (HAdV-41), 293TE cells were infected with purified wild-type HAdV-41, and ultrathin sections of infected cells were prepared and observed under a transmission electron microscope. Results showed that HAdV-41 entered host cells mainly through three ways: non-clathrin-coated pit, clathrin-coated pit, and direct penetration of plasma membrane. In addition, cell microvilli might help HAdV-41 enter cells. After entering into cells, HAdV-41 virus particles could be found in vacuoles or lysosomes or be in a free state in cytoplasm. Only free virus particles could be found near nuclear pores (NP), suggesting that the virus needed to escape from lysosomes for effective infection and viral nucleoprotein entered the nucleus through NP. Progeny viruses were as-sembled in the nucleus. Three types of inclusion bodies, which were termed as fibrillous inclusion body, condense inclusion body, and stripped condense inclusion body, were involved in HAdV-41 morphogenesis. In the late phase of viral replication, the membrane integrity of the infected cells was lost and viral particles were released extracellularly. This study reveals the partial process of HAdV-41 morphogenesis and provides more biological information on HAdV-41.
Adenovirus Infections, Human ; virology ; Adenoviruses, Human ; genetics ; growth & development ; physiology ; ultrastructure ; Cell Membrane ; virology ; Cell Nucleus ; virology ; Humans ; Virus Release ; Virus Replication

OBJECTIVETo investigate the therapeutic mechanism of baicalin on rat brain glioma.

METHODSDeep brain glioma models were established by injection of glioma cell line C6 cells into the brain of Wistar rats. The rats at 7 days after modeling were randomly divided into tumor control group (0.9% NaCl solution 30 mg×kg(-1)×d(-1) gavage)and experimental groups. The experimental rats was divided into 3 groups: low dose group (50 mg×kg(-1)×d(-1)), middle dose group (100 mg×kg(-1)×d(-1)) and high dose group (200 mg×kg(-1)×d(-1)), given the baicalin by gavage. Pathological and electron microscopic changes were observed. The expressions of p53 and Bcl-2 were determined by immunohistochemistry, and the changes of MRI, the average survival time and body weight of the rats in each group after treatments were analyzed.

RESULTSCompared with the control group, the tumor diameter and volume of high dose group rats before sacrifice were significantly reduced (P < 0.01), and the survival time was significantly prolonged (P < 0.01). Immunohistochemistry revealed strong positive expression rate of mutant p53 (84.47 ± 3.74)% and moderately positive rate (47.28 ± 2.38)% in the control group, significantly higher than that in the negative group (12.91 ± 1.07)% (P < 0.01). The positive rate of mutant p53 of the high dose group was (46.42 ± 2.19)%, significantly lower than that of the control group (84.47 ± 3.74)% (P < 0.01). The expression rate of Bcl-2 in the control group was strongly positive (86.51 ± 4.17)% and moderate positive (48.19 ± 2.11)%, significantly higher than that of the negative group (10.36 ± 1.43)% (P < 0.01). Electron microscopy revealed that baicalin caused damages of the cell nuclei and organelles in the gliomas.

CONCLUSIONSBaicalin has significant inhibitory effect on glioma in vivo, and its mechanism may be related to cell apoptosis induced by down-regulated expression of mutant p53, but not related with Bcl-2 expression.

Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Brain Neoplasms ; metabolism ; pathology ; ultrastructure ; Cell Nucleus ; drug effects ; Dose-Response Relationship, Drug ; Down-Regulation ; Flavonoids ; administration & dosage ; pharmacology ; Glioma ; metabolism ; pathology ; ultrastructure ; Magnetic Resonance Imaging ; Male ; Microscopy, Electron, Transmission ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Tumor Burden ; drug effects ; Tumor Suppressor Protein p53 ; metabolism

AIMTo analyse the cytomorphological features of keratinocytes in smears obtained from the oral mucosa of tobacco users and from oral squamous cell carcinoma lesions.

METHODOLOGYOral smears were obtained from clinically, normal appearing mucosa of oral squamous cell carcinoma patients (n=20) and from the mucosa of smokers (n=20), and apparently healthy individuals (n=20) were used as controls. The smears were histochemically stained and cytomorphological assessment of the keratinocytes was carried out. One-way ANOVA (Analysis of Variance) was used for comparing the parameters among multiple groups and Tukey-HSD test was used to compare the mean values between groups.

RESULTSThe mean nuclear area of keratinocytes from the mucosa of tobacco users was 46 +/- 2.57 and that of the oral squamous cell carcinoma lesion was 81.54 +/- 4.31. While there was a significant (P = 0.001) reduction in the cellular area of keratinocytes from oral squamous cell carcinoma lesion when compared with those from oral smears of tobacco users.

CONCLUSIONCytomorphometric analysis of keratinocytes can serve as a useful adjunct in the early diagnosis of oral squamous cell carcinomas.

Azo Compounds ; Carcinoma, Squamous Cell ; pathology ; Cell Nucleus ; ultrastructure ; Cell Size ; Coloring Agents ; Cytodiagnosis ; instrumentation ; methods ; Cytoplasm ; ultrastructure ; Eosine Yellowish-(YS) ; Fluorescent Dyes ; Humans ; Image Processing, Computer-Assisted ; methods ; Keratinocytes ; pathology ; Mouth Mucosa ; pathology ; Mouth Neoplasms ; pathology ; Rosaniline Dyes ; Smoking ; pathology ; Software ; Tobacco, Smokeless

This study was conducted to evaluate the microtubule distribution following control of nuclear remodeling by treatment of bovine somatic cell nuclear transfer (SCNT) embryos with caffeine or roscovitine. Bovine somatic cells were fused to enucleated oocytes treated with either 5 mM caffeine or 150 micrometer roscovitine to control the type of nuclear remodeling. The proportion of embryos that underwent premature chromosome condensation (PCC) was increased by caffeine treatment but was reduced by roscovitine treatment (p < 0.05). The microtubule organization was examined by immunostaining beta- and gamma-tubulins at 15 min, 3 h, and 20 h of fusion using laser scanning confocal microscopy. The gamma-tubulin foci inherited from the donor centrosome were observed in most of the SCNT embryos at 15 min of fusion (91.3%) and most of them did not disappear until 3 h after fusion, regardless of treatment (82.9-87.2%). A significantly high proportion of embryos showing an abnormal chromosome or microtubule distribution was observed in the roscovitine-treated group (40.0%, p < 0.05) compared to the caffeine-treated group (22.1%). In conclusion, PCC is a favorable condition for the normal organization of microtubules, and inhibition of PCC can cause abnormal mitotic division of bovine SCNT embryos by causing microtubule dysfunction.
Animals ; Caffeine/pharmacology ; Cattle/embryology/*physiology ; Cell Nucleus/drug effects/*physiology/ultrastructure ; Female ; Fertilization in Vitro/veterinary ; Male ; Microscopy, Confocal/veterinary ; Microtubules/drug effects/*physiology/ultrastructure ; Nuclear Transfer Techniques/veterinary ; Oocytes/*physiology ; Pregnancy ; Purines/pharmacology

Bone Marrow Cells ; cytology ; Cell Cycle ; Cell Differentiation ; Cell Nucleus ; genetics ; Cells, Cultured ; Diploidy ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Immunophenotyping ; Mesenchymal Stromal Cells ; cytology ; metabolism ; ultrastructure ; Transfection

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

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

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

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