This paper introduces a new method for color blood cell image segmentation based on FCM algorithm. By transforming the original blood microscopic image to indexed image, and by doing the colormap, a fuzzy apparoach to obviating the direct clustering of image pixel values, the quantity of data processing and analysis is enormously compressed. In accordance to the inherent features of color blood cell image, the segmentation process is divided into two stages. (1)confirming the number of clusters and initial cluster centers; (2) altering the distance measuring method by the distance weighting matrix in order to improve the clustering veracity. In this way, the problem of difficult convergence of FCM algorithm is solved, the iteration time of iterative convergence is reduced, the execution time of algarithm is decreased, and the correct segmentation of the components of color blood cell image is implemented.
Algorithms ; Color ; Cytological Techniques ; methods ; Erythrocytes ; ultrastructure ; Humans ; Image Interpretation, Computer-Assisted ; Leukocytes ; 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

Cell patterns in open wound healing are studied by both light and electron microscopic examinations in regards to time sequence, metamorphosis, and functional aspects. Process of the open wound healing clearly exhibited not only time sequence of cllular appearance but also zonation of cells. In the initial stage, until the 3rd day, the neutrophilic polymorphonuclear leukocytes were predominant and particularly concentrated in the scab region. The mononuclear cells were active cells during the 1st to 7th day and were mainly concentrated in the subscab region. The fibroblastic activities started from the 3rd day and became very active during the 5th to the 10th day, and they were concentrated at granulation tissue region. During the process of wound healing, the cellular elements underwent metamorphosis; The neutrophils from normal to swollen and finally degenerating; the mononuclear to macrophages; the fibroblasts from immature to mature actively protein synthesizing cells. The functions of each cellular element can not be determined with certainty. However, the main function of neutrophils in wound healing is likely the formation of front line defense as a part of the scab formation on the surface. And the major function of mononuclear cells is to debride exudates and damaged tissue debris especially at the subscab area and that of the fibroblasts to replace the tissue defect by proliferation and production of fibrous proteins.
Animal ; Epithelium/ultrastructure ; Fibroblasts/ultrastructure ; Leukocytes/ultrastructure ; Rats ; Skin/injuries* ; Skin/pathology ; Wound Healing* ; Wounds, Penetrating/pathology*

OBJECTIVE: To observe changes on cell membrane in blood cells after they were been electrified. METHODS: Blood were electrified for 5, 10, 20, 30 s, 1 min respectively, and Scanning electron microscope was used to detect the changes on their cell membranes. RESULTS: Pores were detected both on electrified erythrocytes and leukocytes with round or ellipse shapes. The erythrocytes often have one or more pores while the leukocytes often have more pores looked like cribble. The rates of perforated cells were increased with the prolonging time of electrification, 5 s with 6% and 1 min increased to 40%. CONCLUSIONS Alternating current can cause the cell perforating, and the rates of perforated cell were increased with the prolonging time of electrification.
Adult ; Blood Cell Count ; Cell Membrane/ultrastructure* ; Cell Membrane Permeability ; Electroporation/methods* ; Erythrocytes/ultrastructure* ; Female ; Humans ; In Vitro Techniques ; Leukocytes/ultrastructure* ; Male ; Microscopy, Electron, Scanning ; Middle Aged ; Young Adult

Karyotypes were prepared from peripheral blood leukocytes in 18 couples with his-tories of habitual abortions. The Standard chromosome analysis and G-banding techniques were studied. The abnormal karyotypes seen were one case with 20% of 45, XX, -14, -15, t(14/15), one case of 46, XY/45, XY, -21 mosaicism, one case of 45, XX, -14, -21, t(14/21), one case of 46, XX/45, XO mosaicism and one case of 46, XYq+. Many other types of chromosomal abnormalities from many reports in couples with spontaneous abortions are discussed.
Abortion, Habitual/genetics* ; Adult ; Chromosome Aberrations* ; Female ; Human ; Leukocytes/ultrastructure ; Male ; Mosaicism ; Pregnancy ; Translocation (Genetics)

The anticancer agent's FT-207, N1-(2'-tetrahydrofuryl)-5-fluorouracil, a derivative of 5FU (5-fluorouracil), induced chromosome damage to the human leukocyte was investigated. FT-207 inhibit mitosis and cause chromatid and chromosome breakage and chromatid exchange with 20 ug/ml for 48 to 72 hours of treatment. However, with 15 ug/ml for 72 hours only delayed spiralization was produced in some of the chromosomes in the same cells. The random distribution of chromosome breakage were observed and the effect of FT-207 on the chromosomes of human leukocytes were time dependent rather than concentration dependent. The comparision of the effect of mitomycin C on human leukocytes and the action of FT-207 at specific times during the cell cycle were discussed.
Cells, Cultured ; Chromosome Aberrations* ; Female ; Fluorouracil/analogs & derivatives* ; Human ; Leukocytes/drug effects ; Leukocytes/ultrastructure* ; Male ; Mitotic Index/drug effects ; Tegafur/pharmacology* ; Time Factors

Chromosome analysis were carried out on peripheral blood leukocytes of breast cancer patient during the irradiation therapy after unilateral simple mastectomy. The observations were made at intervals varying from one to 5 weeks during the therapy and one month after the completion of tile treatment. During the first and second weeks of treatment normal metaphase was noted and during the 4th and 5th weeks, there were no mitotic figures from the cell population. The chromosomal aberrations found after 3 weeks of treatment were, 11% of simple chromatid breaks, 7% of chromatid interchanges (translocations) and 8% of fragments. One month after the completion of the course of treatment showed a return of mitosis and that total chromatid breaks had decreased to 5%. Radiation effects on cell division and chromosome aberration are discussed.
Breast Neoplasms/genetics* ; Breast Neoplasms/radiotherapy ; Breast Neoplasms/surgery ; Carcinoma, Intraductal, Noninfiltrating/genetics* ; Carcinoma, Intraductal, Noninfiltrating/radiotherapy ; Carcinoma, Intraductal, Noninfiltrating/surgery ; Case Report ; Chromosome Aberrations* ; Female ; Human ; Leukocytes/ultrastructure* ; Middle Age

The objective was to explore the feasibility of differentiation of human umbilical cord blood mononuclear cells into endothelial cells induced by cytokines in vitro and to study the possibility of using cord blood stem cells in ischemic diseases therapy. The cells were isolated from umbilical cord blood by using lymphocyte separation solution, and committedly differentiated by using VEGF, bFGF and IGF-I in a liquid culture system. The results showed that the combination of cytokines produced a large number of caudated adherent cells and flow cytometric analysis revealed endothelial marker vWF expressed in about 80% cells, and the endothelial -specific Weibel-Palade body was detected in the cytoplasm by electronic microscope. It is concluded that human umbilical cord blood mononuclear cells may be induced to differentiate into endothelial cells induced by VEGF, bFGF and IGF-I. Human umbilical cord blood MNC may be an ideal source of adult stem cells for the treatment of the ischemic disease.
Cell Differentiation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; ultrastructure ; Fetal Blood ; cytology ; Fibroblast Growth Factor 2 ; pharmacology ; Flow Cytometry ; Humans ; Insulin-Like Growth Factor I ; pharmacology ; Leukocytes, Mononuclear ; cytology ; Microscopy, Electron ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo investigate the promoter methylation of p16, FHIT and RASSF1A gene and telomere damage in the workers exposed to coal tar pitch, and to explore the effective biomarker of occupational exposure to coal tar pitch.

METHODS180 cases of workers exposed to coal tar pitch in a certain carbon plant named as exposure group, and 145 healthy cases with a medical examination in the first affiliated hospital of Zhengzhou University were selected as control group. Relative telomere length in peripheral blood DNA was detected using real-time quantitative PCR, and the promoter methylation rate of p16, RASSF1A and FHIT gene in peripheral blood DNA were determined by real-time quantitative methylation specific PCR. The relative telomere length and gene promoter methylation in two groups were compared, and influencing factors were analyzed.

RESULTSRelative telomere length in exposed group was lower than that in the control group, and the difference was statistically significant (Z = -5.395, P < 0.001). There was no significant difference in the promoter methylation rate of p16, FHIT and RASSF1A gene between the two groups (P > 0.05). Stratification analysis by gender, age, and smoking, we found that when the age was less than or equal to 40, the promoter methylation rate of p16 in exposed group was more than that in control group, and the difference was statistically significant (Z = -1.914, P = 0.011).

CONCLUSIONOccupational exposure to coal tar pitch may induce leukocyte DNA telomere length of human peripheral blood shortened, and may not change the promoter methylation rates of p16, FHIT and RASSF1A gene.

Acid Anhydride Hydrolases ; genetics ; Coal Tar ; adverse effects ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; DNA Methylation ; Humans ; Leukocytes ; drug effects ; Neoplasm Proteins ; genetics ; Occupational Exposure ; adverse effects ; Promoter Regions, Genetic ; Telomere ; drug effects ; ultrastructure ; Tumor Suppressor Proteins ; genetics

By using AdEasy system, which is based on the homologous recombination in bacteria, an EGFP labled recombinant adenovirus vector containing hVEGF(165) was generated quickly and its property was studied in vitro. First, hVEGF(165) coding sequence was subcloned into the shuttle plasmid pAdTrack-CMV, then linearized and cotransferred with adenoviral backbone vector pAdEasy-1 into E. coli strain BJ(5183). After positive kanamycin-resistant colony was picked up, the recombinant adenoviral plasmid was identified by restriction analysis with PacI and transfected into HEK 293 cells to assembly replication-defective adenovirus Ad-EGFP/hVEGF(165). The further amplified recombinant adenoviruses were purified by CsCl banding at 32,000 rpm for 18 to 24 hours. Electron microscopy and PCR were performed for testing the recombinant adenovirus. The results showed that the purified particles were homogenous hexagon with a high titer up to 2 x 10(12)pfu/ml. An amplified band of 540 bp fragment demonstrated the successful insert of hVEGF(165). Under fluorescence microscopy, the expression of EGFP was easily detected in HEK 293 and other target cells. The maximal stimulating effect on the proliferation of hUVEC was obtained when the given multiplicity of infection (MOI) of Ad-EGFP/hVEGF(165) was 100. The rate of EGFP positive mouse bone marrow mononuclear cells analysed by flow cytometry was 27.3% after 24 hour-incubation with Ad-EGFP/hVEGF(165) (MOI = 100), and the expression of hVEGF(165) protein in the conditioned medium was 1385 +/- 332 pg/10(6) cells. It is concluded that the construction of adenovirus vector by homologous recombination in bacteria using AdEasy system can be quickly and easily performed, and the prepared high titer of Ad-EGFP/hVEGF(165) is an efficient helpful vector to transfer genes into target cells, all of which make the further in vivo experiments with VEGF(165) possible.
Adenoviridae ; genetics ; ultrastructure ; Animals ; Cell Division ; genetics ; Cell Line ; Cells, Cultured ; Cloning, Molecular ; methods ; Endothelial Growth Factors ; genetics ; metabolism ; Endothelium, Vascular ; cytology ; metabolism ; Genetic Vectors ; genetics ; ultrastructure ; Green Fluorescent Proteins ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Leukocytes, Mononuclear ; cytology ; metabolism ; Luminescent Proteins ; genetics ; metabolism ; Lymphokines ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Microscopy, Electron ; Microscopy, Fluorescence ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors