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

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

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

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

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*

PURPOSE: Radiation adaptive response in human peripheral lymphocytes and mouse bone marrow cells was investigated using both metaphase analysis and micronucleus assay. We assessed the correlation between both tests. MATERIALS AND METHODS: Two groups of the human peripheral lymphocytes and mouse bone marrow cells were exposed to low dose (conditioning dose, 0.18 Gy) or high dose (challenging dose, 2 Gy) gamma-rays. The other 4 groups were exposed to low dose followed by high dose after several time intervals (4, 7, 12, and 24 hours, respectively). The frequencies of chromosomal aberrations in metphase analysis and micronuclei in micronucleus assay were counted. RESULTS: Chromosomal aberrations and micronuclei of preexposed group were lower than those of the group only exposed to high dose radiation. Maximal reduction in frequencies of chromosomal aberrations were observed in the group to which challenging dose was given at 7 hour after a conditioning dose (p<0.001). Metaphase analysis and micronucleus assay revealed very good correlation in both human lymphocytes and mouse bone marrow cells (r=0.98, p<0.001; r=0.99, p=0.001, respectively). CONCLUSION: Radiation adaptive response could be induced by low dose irradiation in both human lymphocytes and mouse bone marrow cells. There was a significant correlation between metaphase analysis and micronucleus assay
Animals ; Bone Marrow Cells* ; Bone Marrow* ; Chromosome Aberrations ; Cytogenetics* ; Humans* ; Lymphocytes* ; Metaphase* ; Mice* ; Micronucleus Tests

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: 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: Comparative genomic hybridization (CGH) was used to detect any amplified or deleted chromosomal regions in tumors by mapping their locations on normal metaphase chromosomes. METERIALS AND METHODS: Twenty-six gastric carcinomas and their adjacent mucosa were screened for chromosomal aberrations using CGH. RESULTS: All carcinomas had chromosomal aberrations, and chromosomal material was more likely to be gained than lost. Ten out of 26 adjacent mucosa had chromosomal aberrations, and a gain was less frequently observed than a tumor (1.6/2.6). The most common gains were detected on 13q (58.3%), 8q (30.8%), 6q (27.0%), and 20p (19.2%), while the most frequent losses were detected on 17p (38.5%) and 16q (7.2%). The most commonchromosomal aberrations in the adjacent mucosa were a gain of 13q (11.5%) and a loss of 17q (11.5%). The tumors had more chromosomal gains of 2q, 3q, and 13q and more losses of 17p and 16q than the adjacent mucosa. CONCLUSIONS: The most common gain in the tumors was detected on 13q, 8q, 6q, and 20p, and the most frequent loss was on 17p and 16q. While CGH may be useful in predicting the prognosis or therapeutic decision of gastric carcinomas, further study of several candidate genes, such as DP1, FLT1, c-myb, AIB1, BTAK, is needed to clarify gastric carcinogenesis and its progression.
Carcinogenesis ; Chromosome Aberrations ; Comparative Genomic Hybridization* ; Metaphase ; Mucous Membrane* ; Prognosis ; Stomach Neoplasms

This experiment was undertaken in order to know the effect of leucocytes on the maturation of mouse oocytes in vitro. Leucocytes obtained from heart puncture of mouse (3 X 10(4) cells/mm3) inhibited the maturation of mouse oocytes. The egg toxic activity declined with decreasing leucocyte concentration. It was found that egg toxic effect of leucocytes is not species specific. The activity of intact leucocytes or equal numbers of leucocytes that were destroyed was similar and which seems not to be influenced by the physiological stats of leucocytes.
Animal ; Culture Media ; Female ; Leukocytes* ; Metaphase ; Mice ; Oocytes/cytology* ; Ovum/cytology*