We report here several experiences of interphase cytogenetics, using fluorescence in situ hybridization (FISH) technique, for the detection of chromosome aberrations. FISH, using alpha satellite specific probes of 18, X, Y chromosomes, was done in interphase nuclei from peripheral blood of patients with Edwards' syndrome, Klinefelter's syndrome and Turner's syndrome with healthy male and female controls, respectively. The distributions of fluorescent signals in 100 interphase nuclei were well correlated with metaphase findings. Nowadays FISH plays an increasingly important role in a variety of research areas, including cytogenetics, prenatal diagnosis, tumor biology, gene amplification and gene mapping.
Adolescent ; Adult ; Cell Nucleus/*ultrastructure ; Child ; Chromosome Aberrations/*physiology ; Chromosome Banding ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Infant, Newborn ; Interphase/*physiology ; Male

Partial hepatectomy (PH) endorses quiescent hepatocytes to reenter the cell cycle. The regenerating liver returns to its preresection weight after 7 days, following one or two cell division and maintains nearly its original volume after then. We focused on the inhibition of further hepatocyte proliferation, hypothesizing possible involvement of cell cycle upregulators and inhibitors. We studied protein levels in expression of cyclins, cyclin dependent kinases (CDKs) and CDK inhibitors (CKIs), and their in situ hepatic lobular distributions in partial hepatectomized rat liver. Cyclin E was expressed in the same levels in normal liver and after PH. Expression of cyclin A, not detected in normal liver, increased in following times after PH and reached a maximum at 7 day. CDK2 and 4 showed increased expression toward terminal period. Contradictory findings of cyclin A and these CDKs might play an important role in the inhibition of further cell division, although still unclear. Constitutively expressed CDK6 decreased after 1 day. p18 showed peak expression within 1 day, and p16, p21, p27 and p57 were stronger at terminal periods. During the expected period of their activity, intranuclear translocations were observed in cyclin E, p18 and p16. There was no evidence of regional distribution in hepatic lobular architecture, instead, diffuse in situ expression, corroborating synchronous event, was found.
Animal ; Cell Cycle/physiology* ; Cyclin-Dependent Kinases/metabolism ; Cyclin-Dependent Kinases/antagonists & inhibitors ; Cyclins/metabolism* ; Cyclins/immunology ; Flow Cytometry ; Hepatectomy ; Immunoblotting ; Immunohistochemistry ; Interphase/physiology ; Liver/metabolism* ; Liver Regeneration/physiology* ; Male ; Rats ; Rats, Sprague-Dawley ; S Phase/physiology

Bromodeoxyuridine, an analogue of thymidine, can be detected by means of monoclonal antibodies and utilized as a marker of the S-phase of the cell cycle. In vitro immunohistochemical application of the BrdU/anti-BrdU-MAb method permits a quantitative assessment of the proliferative activity of a tissue as well as the direct location of the actively replicating cells in histological sections. In this paper, a method for the detection of the labeling index of S-phase cells in normal and neoplastic tissues with in vitro BrdU labeling and standard immunohistochemical techniques using anti-BrdU-MAb and avidin-biotin peroxidase complex is described. We have employed this method in 47 human solid tumor samples, including squamous cell carcinomas of head and neck and cervix uteri, adenocarcinomas and malignant lymphomas, and also evaluated the possible application of the BrdU labeling index to estimate the cycling S-phase cells in neoplastic cell populations. In our data, the in vitro labeling index varied greatly in an individual case (3.56-29.2%) and from an area to an area within the same case. Squamous cell carcinomas of the head and neck showed higher LI than those of the cervix uteri. A case of metastatic carcinoma to the lung from ductal carcinoma of the breast had the highest LI (29.2%), in contrast to the low LI (3.6%) in the primary ductal carcinoma of breast.
Adenocarcinoma/immunology/pathology ; Antibodies, Monoclonal/*immunology ; Breast Neoplasms/immunology/pathology ; Bromodeoxyuridine/*immunology ; Carcinoma, Squamous Cell/immunology/pathology ; Cell Nucleus/immunology/*physiology ; Head and Neck Neoplasms/immunology/pathology ; Humans ; Immunohistochemistry ; *Interphase ; Lymphoma/immunology/pathology ; Neoplasms/immunology/*pathology

OBJECTIVECompare two methods of preparing human unfertilized oocytes interphase nucleus and analyze the relationship between the different in vitro fertilization(IVF) indications, ovarian stimulation protocols, women's age and frequency of chromosome 17 aneuploidy.

METHODSTarkowski's air-drying method(3:1 methanol:acetic acid) and Coonen's 0.1% Tween 20/0.01 mol/L HCl method were used to fix human unfertilized oocytes interphase nucleus, and telomeric probe of 17qter was used by standard fluorescence in situ hybridization(FISH) procedures to confirm chromosome 17qter aneuploidy.

RESULTSOf 36 human unfertilized oocytes, 24 were haploid (66.7%), 7 were disomic (19.44%), 5 were trisomic (13.89%). The overall frequency of aneuploidy was 33.3%. There were no differences between the protocols characterized by different maternal age, IVF indication, ovarian stimulation.

CONCLUSIONTarkowski's air dry method is as good as the method of Coonen's, but the latter method can avoid the smell pollution of the methanol and acetic acid, and it is easy to operate. The chromosome 17 aneuploidy is one of the factors to cause in vitro fertilization failure of human oocytes.

Adult ; Aneuploidy ; Cell Nucleus ; genetics ; ultrastructure ; Chromosomes, Human, Pair 17 ; genetics ; ultrastructure ; DNA Probes ; Diploidy ; Embryo Transfer ; Female ; Fertilization in Vitro ; methods ; Haploidy ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Interphase ; genetics ; Karyotyping ; Microscopy, Fluorescence ; methods ; Ovulation Induction ; methods ; Ovum ; physiology ; Trisomy