No abstract available.
Animals ; Metaphase* ; Mice* ; Oocytes*

The task for chromosome karyotyping and diagnosis is requiring repetitive, time consuming job and high cost even it is done by well-experienced cytogenetists. Therefore an web based chromosome karyotyping instruction system has been established to be able to analyze chromosomes and obtain necessary advises from the database instead of human experts and the database is including 2 divisions with database and agent.For the first of all, database model was constructed with relational database consisting of Patient_DB, image_DB, Disease_DB and Manage_DB. As the second procedure, knowledge base by IF THEN production rule was implemented to a knowledge domain with normal and abnormal chromosomes. For the last, independent agent with the inference by knowledge base could enter the inference data into the database.Experimental data were composed of normal chromosomes of 2,736 patients' cases and abnormal chromosomes of 259 patients' cases that have been obtained from GTG-banding metaphase peripheral blood and amniotic fluid samples.The completed system provides variously morphological information by analysis of normal or abnormal chromosomes and it also makes users enable to control and search the information in a short period with learning of high amount of knowledge.
Amniotic Fluid ; Diagnosis ; Female ; Humans ; Karyotyping* ; Knowledge Bases ; Learning ; Metaphase

Chromosome analysis has been widely used in clinics including prenatal diagnosis. To obtain high-quality metaphase chromosomes, researchers have attempted to modify the methods for chromosome culture, preparation and analysis. Some large research centers also tried to establish standards for quality control. In this paper, modification of methods for the preparation of chromosomes in the last decade is reviewed.
Cells, Cultured ; Chromosomes, Human ; Cytogenetic Analysis ; Humans ; Karyotyping ; Metaphase

Oocyte cryopreservation is widely used for clinical and social reasons. Previous studies have demonstrated that conventional slow-freezing cryopreservation procedures, but not storage time, can alter the gene expression profiles of frozen oocytes. Whether vitrification procedures and the related frozen storage durations have any effects on the transcriptomes of human metaphase II oocytes remain unknown. Four women (30-32 years old) who had undergone IVF treatment were recruited for this study. RNA-Seq profiles of 3 fresh oocytes and 13 surviving vitrified-thawed oocytes (3, 3, 4, and 3 oocytes were cryostored for 1,2, 3, and 12 months) were analyzed at a single-cell resolution. A total of 1987 genes were differentially expressed in the 13 vitrified-thawed oocytes. However, no differentially expressed genes were found between any two groups among the 1-, 2-, 3-, and 12-month storage groups. Further analysis revealed that the aberrant genes in the vitrified oocytes were closely related to oogenesis and development. Our findings indicated that the effects of vitrification on the transcriptomes of mature human oocytes are induced by the procedure itself, suggesting that long-term cryostorage of human oocytes is safe.
Adult ; Cryopreservation ; Female ; Humans ; Metaphase ; Oocytes ; RNA-Seq ; Vitrification

PURPOSE: Because of difficulty of obtaining metaphase cells from tumor specimens, there are only a few cytogenetic studies in nasal NK/T-cell lymphomas, and so far no consistent specific chromosomal abnormalities have been described. In this study, we have used degenerate oligonucleotide primed PCR (DOP-PCR) and comparative genomic hybridization (CGH) to deter mine chromosomal alterations from 6 nasal NK/T-cell lymphoma tissues dissected from formalin- fixed paraffin-embedded slide sections. MATERIALS AND METHODS: For the isolation of tumor DNA, four 7-micrometer-thick tissue sections from each sample were dewaxed and rehydrated, and areas of high tumor cell content (more than 60%) were dissected and pooled into a tube. Normal DNA was prepared from the peripheral blood of a healthy volunteer. Tumor DNA was labeled with biotin-16-dUTP by DOP-PCR and normal DNA was labeled with digoxigenin-dUTP using a nick translation kit. In CGH, equal amounts of differently labeled DNA from the tumors and normal reference DNA were hybridized simul taneously to normal metaphase chromosomes. They were visualized by different fluordegrees Chromes, and the signal intensities were quantitated separately as gray levels for each chromosome. The over- and underrepresented DNA segments were determined by computation of image ratios and average ratio profiles. RESULTS: Our results show that gains of DNA copy number were more prevalence than DNA losses. The most commonly observed gains were mapped to chromosomal regions of 1p32.2 ter,19 and 20 in 4 of 6 cases (67%). The other frequent gains were found on chromosomes 12q in 3 of 6 cases. The most frequent loss was detected on 6q in 4 of 6 cases(67%), and less fre quently observed on 13q21.1 q34 and 13q14 q34. CONCLUSION: These genomic changes found in specific chromosomal regions are likely to harbor genes of importance in nasal NK/T-cell lymphomagenesis, therefore such cytogenetic mapping of genomic imbalance may be of value for further molecular delineation of NK/T-cell lymphoma.
Chromosome Aberrations ; Comparative Genomic Hybridization* ; Cytogenetics ; DNA ; Healthy Volunteers ; Lymphoma* ; Metaphase ; Polymerase Chain Reaction ; Prevalence

BACKGROUND: The KCl hypotonic treatment is important for swelling the cells and adequate spreading of chromosomes on the slide. Cytogenetic laboratory usually use 0.075M KCl solution. Sometimes, it is difficult to obtain enough and good quality of metaphase cells, because of inadequate hypotonic treatment. The purpose of this study was to evaluate the mitotic index and band resolution according to the different KCl concentration. METHODS: The group I included blood specimens obtained from 14 newborns (median age 1 day, range 1-8 days) and 4 cord blood. The group II included 16 persons whose median age was 28 years (1-37 years). The blood was cultured in RPMI 1640 medium with fetal calf serum and phytohemagglutinin for 72 hours. Mitosis was arrested by adding colcemid (100 ng/mL). The hypotonic treatment was done by adding different KCl concentration such as 0.075M, 0.068M and 0.057M for 30 minutes at 37 degrees C. The mitotic index was calculated as the number of metaphase cells per total 1,000 cells. The band resolution was evaluated by 2 persons independently. RESULTS: For group I, the mitotic index was not different according to the KCl concentration; 0.075M, 18.8 (5.5~31.5); 0.068M, 22.3 (11~32.5); 0.057M, 20.5 (2.5~29), (P=0.137). The proportion of cells with 400 or more band resolution was significantly higher in specimens treated with 0.068M KCl than those treated with 0.075M KCl; 0.075M, 67.8% (56~92.5); 0.068M, 73.6% (46.1~84.6); 0.057M, 71.6% (63~89.2), (P=0.027). For group II, the results were similar to those of group I. The mitotic index was as follows; 0.075M, 22.3 (5~28); 0.068M, 26 (4~34.5); 0.057M, 21.5 (2.5~36.5), (P=0.568). The proportion of cells with 400 or more band resolution was as follows; 0.075M, 66.6% (42.8~83.3); 0.068M, 69.7% (54.3~87.5); 0.057M, 68.2% (50~78.6) (P=0.04). CONCLUSIONS: For 0.068M or 0.057M KCl treatment, band resolution was improved, while the mitotic index was similar to that of 0.075M KCl. We suggest use of 0.068M or 0.057M KCl hypotonic treatment in addition to 0.075M KCl for chromosome preparation of peripheral blood.
Cytogenetics ; Demecolcine ; Fetal Blood ; Humans ; Infant, Newborn ; Metaphase ; Mitosis ; Mitotic Index*

Sister chromatid exchanges(SCEs) and cell cycle kinetics were proposed as a sensitive and quantitative assay for mutagenicity and cytotoxicity in short-term cultures of phytohemagglutinin(PHA)-stimulated human lymphocytes. Therefore, this study was performed to investigate the relation between the cytotoxic effects and sister chromatid exchanges. The results are summarized as follows: 1) The frequency of SCEs per cell are 13.1+/-2.8 in the lower concentration of 6.25x10(-9) M and 75.8+/-8.2 in the highest concentration of 1.00+/-10(-7) M. Mitotic index is decreased in the higher concentration of mitomycin C. The result indicates that mitomycin C led to a dose dependent increase in SCE frequency, but decrease in mitotic index. 2) Chromosomal analysis was performed on metaphase cells that have divided one, two, and three or more times for cell cycle kinetics by fluorescence-plus-Giemsa(FPG) technique. According to the increased but the cells of third division are greatly decreased. 3) The frequency of SCEs per chromosome by chromocomal group are decreased gradually from A group to G group. But relationships between specific chromosomal group and SCEs frequency are not found.
Cell Cycle ; Chromatids ; Humans ; Humans* ; Kinetics ; Lymphocytes ; Metaphase ; Mitomycin* ; Mitotic Index ; Siblings* ; Sister Chromatid Exchange*

OBJECTIVE: The purpose of this study was to evaluate the efficacy and effect of various cryopreservation method on the survival and the cytoskeletal stability of metaphase II mouse oocyte. METHODS: Mouse ovulated oocytes were collected and cryopreserved by a modified slow-freezing method with 1.5 M 1,2-propanediol (PrOH)+0.1 M sucrose or by vitrification using cryo loop and EM grid with 40% ethylene glycol+0.6 M sucrose. Four hours after thawing, intact oocytes were fixed and stained with fluorescein isothiocyanate (FITC)-conjugated monoclonal anti-beta-tubulin antibody to visualize spindle and propidium iodide (PI) to visualize chromosome. Spindle morphology was classified as follows: normal (barrel-shaped), slightly and absolute abnormal (multipolar or absent). RESULTS: Survival rate of the frozen-thawed oocytes in vitrification group was significantly higher than that of slow-freezing group (62.7% vs. 24.4%, p<0.01). Vitrification with cryo loop showed significantly higher survival rate than that with EM grid (67.7% vs. 53.5%, p<0.05). On the other hand, proportion of normal spindle and chromosome configurations of the frozen-thawed oocytes between two vitrification group was not significantly different. CONCLUSION: For mouse ovulated oocytes, vitrification with cryo loop may be a preferable procedure compared to slow-freezing method. Further study should be needed to investigate developmental competency of frozen-thawed mouse oocytes.
Animals ; Cryopreservation* ; Fluorescein ; Hand ; Metaphase ; Mice* ; Oocytes* ; Propidium ; Propylene Glycol ; Sucrose ; Survival Rate ; Vitrification

PURPOSE: The purpose of this study was to develop in vivo dosimetries using both chromosomal aberrations and micronuclei in mice to assess biological effects of radiations. MATERIALS AND METHODS: Five each mice were irradiated with 0, 1, 2, 3, 4, 5, 10 Gy of Cs-137 gamma-rays. We scored numbers of chromosomal aberrations in metaphase spreads and numbers of micronuclei in bone marrow smears under light microscope, and obtained the dose-response relationships. We also examined the relationship between the two dose-response curves. RESULTS: The frequency of both chromosomal aberrations and micronuclei increased with dose, in a linear-quadratic manner. The delta, beta, and alpha coefficients were 0.0176, 0.0324, and 0.0567 for metaphase analysis (r=1.0, p<0.001) and 0.0019, 0.0073, and 0.0506 for micronuclei assay (r=1.0, p<0.001). The frequency of chromosomal aberrations and micronuclei in diffirent radiation doses was significantly correlated (r=0.99, p<0.01). CONCLUSION: In vivo dosimetry using either metaphase analysis or micronucleus assay was feasible in mice. These methods could be useful to evaluate biological effects of radiation.
Animals ; Bone Marrow Cells* ; Bone Marrow* ; Chromosome Aberrations ; Metaphase* ; Mice* ; Micronucleus Tests

OBJECTIVE: To investigate the effects of female age on in vitro maturation and fertilization of immature oocytes from controlled ovarian hyperstimulation (COH) in human IVF-ET program. METHOD: A total of 96 immature oocytes (GV & metaphase I) obtained from 40 cycles of IVF-ET (29 patients). The mean age of female patients was 31.8+/-3.1 years. Ovulation was triggered by urinary or recombinant hCG. Immature oocytes were cultured with YS medium containing 30% of patients' human follicular fluids, LH (1 IU/mL), FSH (1 IU/mL) and EGF (10 ng/mL), and then matured oocytes were fertilized by ICSI. In vitro maturation and fertilization of immature oocytes were analyzed according to age of female (< 34 or > or = 34 years). RESULTS: The maturation rate was similar between two groups (68% vs 64%). The fertilization rate of in?vitro-matured oocytes was higher in patients < 34 years old, but there was no statistical significance (64% vs 50%, p=0.347). The fertilization rate of in-vitro-matured oocytes was significantly lower compared with those of in-vivo-matured oocytes in both age groups (64% vs 79%, p=0.035, 50% vs 86%, p=0.007). CONCLUSION: In older female group, fertilization rate of in-vitro-matured oocytes seems to be decreased. Further investigations should be warranted to increase fertilization potential of in-vitro-matured oocytes.
Adult ; Epidermal Growth Factor ; Female ; Fertilization* ; Follicular Fluid ; Humans* ; Metaphase ; Oocytes* ; Ovulation ; Sperm Injections, Intracytoplasmic