OBJECTIVETo establish a multicolor primed in situ labeling (PRINS) protocol for chromosome detection in uncultured amniocytes.

METHODSChromosomes 18, X and Y in uncultured amniocytes were simultaneously detected by using the non-ddNTP-blocking multicolor PRINS procedure.

RESULTSWithin 7 h, the 3 chromosomes were simultaneously marked in the same uncultured amniocyte. The chromosome signals were successfully detected in 69 uncultured samples of amniotic fluid. The results were consistent with that obtained by chromosomes in cultured amniocytes.

CONCLUSIONThis multicolor protocol was high throughput, fast, simple, sensitive and reliable in diagnosing chromosome abnormalities in uncultured amniocytes.

Adolescent ; Adult ; Amniotic Fluid ; chemistry ; cytology ; Cells, Cultured ; Chromosomes, Human, Pair 18 ; chemistry ; genetics ; Chromosomes, Human, X ; chemistry ; genetics ; Chromosomes, Human, Y ; chemistry ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Pregnancy ; Prenatal Diagnosis ; methods ; Primed In Situ Labeling ; methods ; Young Adult

Chromosomal translocations of the human mixed-lineage leukemia (MLL) gene have been analyzed for more than 20 yr at the molecular level. So far, we have collected about 80 direct MLL fusions (MLL-X alleles) and about 120 reciprocal MLL fusions (X-MLL alleles). The reason for the higher amount of reciprocal MLL fusions is that the excess is caused by 3-way translocations with known direct fusion partners. This review is aiming to propose a solution for an obvious problem, namely why so many and completely different MLL fusion alleles are always leading to the same leukemia phenotypes (ALL, AML, or MLL). This review is aiming to explain the molecular consequences of MLL translocations, and secondly, the contribution of the different fusion partners. A new hypothesis will be posed that can be used for future research, aiming to find new avenues for the treatment of this particular leukemia entity.
Alleles ; Chromosomes, Human, X ; Epigenesis, Genetic ; Humans ; Leukemia/classification/*genetics/pathology ; Myeloid-Lymphoid Leukemia Protein/chemistry/genetics ; Protein Structure, Tertiary ; Translocation, Genetic

Pelizaeus-Merzbacher disease (PMD) is a rare X-linked recessive disorder with a prototype of a dysmyelinating leukodystrophy that is caused by a mutation in the proteolipid protein 1 (PLP1) gene on the long arm of the X chromosome in band Xq22. This mutation results in abnormal expression or production of PLP. We here present a Korean boy with spastic quadriplegia, horizontal nystagmus, saccadic gaze, intentional tremor, head titubation, ataxia, and developmental delay. The brain magnetic resonance imaging (MRI) showed abnormally high signal intensities in the white matter tract, including a subcortical U fiber on the T2-weighted and fluid attenuated inversion recovery (FLAIR) image. The chromosomal analysis was normal; however, duplication of the PLP1 gene in chromosome Xq22 was detected when the multiplex ligation-dependent probe amplification (MLPA) method was used. We also investigated the pedigree for a genetic study related to PMD. This case suggests that the duplication mutation of the PLP1 gene in patients with PMD results in a mild clinical form of the disorder that mimics the spastic quadriplegia of cerebral palsy.
Brain/*pathology ; Child, Preschool ; Chromosome Mapping ; Chromosomes, Human, X ; Developmental Disabilities/diagnosis/genetics ; Exons ; Gene Duplication ; Humans ; Korea ; Magnetic Resonance Imaging/methods ; Mutation ; Myelin Proteolipid Protein/genetics ; Myelin Sheath/chemistry ; Pelizaeus-Merzbacher Disease/*diagnosis/*genetics ; Polymerase Chain Reaction/*methods

OBJECTIVETo identify the mutation of spondyloepiphyseal dysplasia tarda (SEDL) gene in a large Chinese family with X-linked spondyloepiphyseal dysplasia tarda and to make a discussion on the pathogenesis of SEDL at the molecular level.

METHODSIn two patients, four exons comprising the SEDL open reading frame as well as their exon/intron boundaries were analyzed by bi-directional direct sequencing of PCR products. The sequencing results were compared against the normal sequences in GenBank to find the mutation. Then the mutation was identified in other members of the family.

RESULTSA nucleotide substitution of the splice acceptor in SEDL intron 2, IVS2 -2A-->C,was detected in two affected individuals (IV(15) V(3)) in the Chinese family with SEDL, but no sequence change occurring on exons 3-6 was detected. The transversion was also identified in four heterozygous carriers. The mutation was not found in two unaffected male individuals and fifteen normal controls. Furthermore, four potential carriers were identified in the family.

CONCLUSIONThe mutation IVS2 -2A-->C of SEDL gene was firstly determined in the world. The change of the splice acceptor in SEDL intron 2 may cause skipping of exon 3 which is responsible for the disease. Molecular diagnosis can be made by detecting the mutation.

Alternative Splicing ; genetics ; Base Sequence ; Carrier Proteins ; genetics ; China ; Chromosomes, Human, X ; genetics ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Family Health ; Female ; Genetic Linkage ; Humans ; Male ; Membrane Transport Proteins ; Mutation ; Osteochondrodysplasias ; genetics ; pathology ; Pedigree ; Transcription Factors

PURPOSE: Xp11.2 translocation renal cell carcinoma (RCC) is characterized by various translocations of the TFE3 transcription factor gene. These rare cancers occur predominantly in children and young adults. Here, we review the clinicopathological features of Xp11.2 translocation RCC. MATERIALS AND METHODS: We identified 21 patients with Xp11.2 translocation RCC. We retrospectively analyzed patient characteristics, clinical manifestations, and specific pathological features to assess definitive diagnosis, surgical and systemic treatments, and clinical outcomes. RESULTS: The mean age at diagnosis was 43.4+/-20.0 years (range, 8-80 years; 8 males and 13 females). Eleven patients were incidentally diagnosed, nine patients presented with local symptoms, and one patient presented with systemic symptoms. The mean tumor size was 6.2+/-3.8 cm (range, 1.9-14 cm). At the time of diagnosis, 11, 1, and 5 patients showed stage I, II, and III, respectively. Four patients showed distant metastasis. At analysis, 15 patients were disease-free after a median follow-up period of 30.0 months. Four patients received target therapy but not effectively. CONCLUSIONS: Xp11 translocation RCC tends to develop in young patients with lymph node metastasis. Targeted therapy did not effectively treat our patients. Surgery is the only effective therapy for Xp11 translocation RCC, and further studies are needed to assess systemic therapy and long-term prognosis.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/*genetics ; Biomarkers ; Carcinoma, Renal Cell/diagnosis/*genetics ; Child ; Chromosomes, Human, X/*chemistry ; Female ; Humans ; Kidney Neoplasms/diagnosis/*genetics ; Lymphatic Metastasis ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Translocation, Genetic ; Young Adult