ABSTRACT
Existence of Y derived chromosome in Turner patients is significant due to the risk of gonadoblastoma development, but cytogenetic analysis may fail to detect low levels of Y chromosomal materials. Recent studies using PCR based methods showed higher sensitivity to detect Y-specific sequences, in patients who were Y chromosome-negative cytogenetically. In this study PCR was performed on 44 Turner patients with no Y chromosome by cytogenetic analysis to detect the SRY, AMELY, ZFY, and DYZ1 sequences. Of seven patients whose karyotypes were 45,X/46,X,+mar, three patients were positive for SRY, ZFY, and AMELY. DYZ1 sequences was negative in them. And any of SRY, ZFY, AMELY, and DYZ1 sequences was detected in the remaining 37 patients. This result shows that PCR analysis for Y-specific sequences in Turner patients, especially in patients who have marker chromosome is a significant effort.
Subject(s)
Humans , Cytogenetic Analysis , Genes, sry , Gonadoblastoma , Karyotype , Polymerase Chain Reaction , Turner Syndrome , Y ChromosomeABSTRACT
Androgen insensitivity syndrome (AIS) is a X-linked disorder of sexual differentiation resulting from defective androgen receptor (AR) function. Androgens are secreted by the testes of 46,XY individuals, but there is loss of target organ response to the hormone. The abnormalities of AR are due to defects in the AR gene, and many mutations causing AIS have been reported since the cloning of AR gene. In this study, we analyzed the AR genes in twelve Korean patients with androgen insensitivity syndrome: 9 patients with complete AIS and 3 patients with partial AIS DNAs were isolated from patients with AIS, and the coding region of AR gene was amplified by a polymerase chain reaction using 7 pairs of primers (exon B-H). Sequence analysis of the AR gene was performed using direct sequencing and single strand conformational polymorphism (SSCP). The AR gene mutations were identified in 7 out of 12 patients: 6 of 9 patients with complete AIS, and one of 3 patients with partial AIS. Mutations found were as follows: the point mutation (ATT->ACT) at position 680 of exon D, point mutation (TGG->TGC) at position 751 of exon E, point mutation (CAA->TAA) at position 792 of exon F, point mutations (CGC->TGC, GTG->ATG) at position 855 and 866 of exon G, and the deletion of 13 nucleotides (CGTATCATTGCAT) at position 840 of exon G, respectively. To the best of our knowledge, the point mutations found in exon D, exon E, and exon F, and the deletion in exon G have not been observed before. SSCP revealed bands with abnormal mobility in 10 out of 12 patients tested. Mutations were found 5 out of these 10 patients. The other two patients showed no abnormal band on SSCP, but showed mutations by direct sequencing. In conclusion, we have demonstrated the AR gene mutations, including three novel mutations, in Korean patients with AIS, and these abnormalities might be related to the pathogenesis of androgen insensitivity syndrome.
Subject(s)
Humans , Male , Androgen-Insensitivity Syndrome , Androgens , Clinical Coding , Clone Cells , Cloning, Organism , DNA , Exons , Nucleotides , Point Mutation , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Receptors, Androgen , Sequence Analysis , Sex Differentiation , TestisABSTRACT
Fragile X syndrome is the most common cause of inherited mental retardation. It accounts for 0.2% - 2.7% of patients with mental retardation, based upon the molecular genetic diagnosis. However, the exact prevalence of fragile X syndrome in Korean patients with mental retardation is unknown. We have performed cytogenetic and molecular analysis for fragile X syndrome in 212 Korean patients with mental retardation. Among them, six patients (2.8%) was identified as carrying fragile X syndrome by both cytogenetic and molecular analysis. The results by cytogenetic analysis was identical to those by molecular analysis. Cytogenetic analysis of 6 carriers (mothers of patients with proven fragile X syndrome) showed a fragile X chromosome in one patients (16.7%) while molecular analysis revealed premutation in all patients. PCR method using Klentaq1 Pfu polymerase showed the same results as those by PCR method using Exo(-) Pfu polymerase, but the former method is recommended because of its simplicity in technical aspect. These data suggest that the prevalence of fragile X syndrome in Korean patients with mental retardation is 2.8%, not significantly different from those in Caucasians.
Subject(s)
Humans , Cytogenetic Analysis , Cytogenetics , Diagnosis , Fragile X Syndrome , Incidence , Intellectual Disability , Molecular Biology , Polymerase Chain Reaction , Prevalence , X ChromosomeABSTRACT
BACKGROUND: Bone age measurements have clinical significance in estimation of growth status and prediction of final adult height. Mostly used methods of bone age measurements are Tanner Whitehouse method(TW2) and Greulich-Pyle method(OP). TW2 is known to be more accurate method in determining the bone age, compared to GP. But GP is being used more widely despite some shortcomings, because TW2 is time consuming and need special training. In this study, we observed the correlation between GP and TW2 to evaluate which bone age among three portions of hand and wrist[metacarpals and phalanges(GP1), carpal bones(GP2), distai radius and ulna (GP3)], measured by GP, was more correlated with the bone age, measured by TW2. METHODS: Left hand/wrist radiographs were taken from 100 prepubertal children with normal growth. These radiogrphs were reviewed by two pediatric endocrinologists independently. Bone ages using TW2 were measured at first, and then GP1, GP2, and GP3 were measured. These bone ages had been compared with TW2, using SAS computer program. RESULTS: The mean chronological age of 100 children was 10.0+/-2.5 years(5 years to 14.7 years range, 63 males and 37 females). The bone age by TW2 was 9.0+/- 2.6 years(2.3 to 13.6 years). The bone age by GP1, GP2, and GP3 were 8.8+/-2.5 years, 8.7+/-2.9 years, and 8.3+/-2.8 years, respectively. Bone ages by TW2 were significantly closer to the chronological age than those by GP. The Pea~rson correlation coefficients of GP1, GP2, and GP3 in eomparison to TW2 were 0,87(p=0.0001), 0.94(p=0.0001), and 0.91(p=0.0001), respectively, There are significant correlatkm between bone ages by TW2 and GP. Bone ages by GP2 and GP3 were statistically significantly different from those by TW2(P<0.01). Bone ages by GP1 has no statistical difference with that by TW2(P=0.64). CONCLUSION: TW2 method is more accurate than GP method in determining the bone age, but it needs time-consuming and laborious efforts. We suggest that the use of GP method for the metacarpals and phalanges can result in a considerable saving of time with no significant loss of accuracy and reproducibility.
Subject(s)
Adult , Child , Humans , Male , Age Determination by Skeleton , Hand , Metacarpal Bones , Radius , UlnaABSTRACT
BACKGROUND: Recently authentic human growth hormone(hGH) has produced in the E coli K-12, W3110 by recombinant DNA tecbnology in Korea In this paper, the clinical efficacy and immunogenicity of this GH was shdied in 38 children with growth hormone deficiency during therapy of 1 year. METHODS: The subjects of this study were aged 4.9-13.9 years, diagnosed by failure of plasma GH to respond to insulin-induced hypoglycemia, arginine and/or L-dopa loading and height below -2 standard deviation of mean for their chronological age. Each patient received GH 0.5-0.7IU/kg/week subcutaneously in 6-7 divided doses. During treatment, vital signs, height, body weight and bone age were checked every 3 months. Complete blood count, urinalysis, blood chemistry and thyroid hormone were checked before and every 6 months. The measurement of serum IGF-1 level and antibody against hGH were performed before and every 6 months during therapy of I year. RESULT: The height velocities significantly increased from 3.3 +/- 1.5cm/year to 10.1 +/- 2.5 and 9.0 +/- 1.8cm/year at 6 and 12 months of therapy, respectively. The height standard deviation score for chronological age were significantly improved from -2.141.50 to -1.74 +/- 1.43 and -1.54 +/- 1.38 at 6 and 12 months of therapy with increasing ratio of bone age to chronological age from 0.72 +/- 0.15 at pretreatment to 0.76 +/- 0.15 at 6 month, 0.79 +/- 0.16 at 12 month of therapy. The plasma IGF-1 level significantly increased during treatment. One of 36 patients(2.8%) showed positive antibody against hGH after 1 year of treatment. During therapy of 1 year, unwanted and remarkable clinical side effect were not observed in all subjects. CONCLUSION: These results indicate that this E. coli derived authentic recombinant growth hormone is very effective in stimulating linear growth in children with growth hormone deficiency.