Congenital adrenal hyperplasia, especially due to steroid-12-hydroxylase(P450c21) deficiency, is one of the most common autosomal recessive inborn errors at adrenal steroidogenesis in Korean. Molecular genetic analysis has demonstrated that there are two steroid 21-hydroxylase genes, CYP21A1P and CYP21A2. The CYP21A2 gene encodes P450c21, whereas the CYP21A1P gene is a pseudogene. Since there is 98 percent homology between the CYP21A1P and CYP21A2 gene in nucleotide sequences, it has hampered the characterization of molecular defects in the CYP21A2 gene.In this study, efforts have been made to selectively PCR amplify the CYP21A2 gene and test feasibility of DNA microextraction from Guthrie card for prospective use of molecular screening. This study was also aimed at investigating deletion mutations in P450c21 deficient patients, as well as allele frequencies and average heterozygosity of exon 1 A/C polymorphism in Korean newborns. Genomic DNAs were obtained from Guthrie cards of 50 Korean newborns by microextraction method and these DNAs were analyzed by PCR-allele specific oligonucleotide(ASO) hybridization. First part of the CYP21A2 gene has been successfully amplified and digested by restriction enzyme using Taq I or Kpn I, subsequently run on 1.5% agarose gel to confirm its specificity. The anterior 1141 bp PCR product was utilized to examine the frequency and average heterozygosity of exon 1 A/C polymorphism in 100 alleles by ASO dot blot hybridization. Amplified genomic DNAs from four P450c21 deficient patients out of three families were screened by PCR to see if any one has complete deletion of the CYP21A2 gene.The results were as follows;1) The average 1230ng of genomic DNA was obtained form single semi-circled Guthrie card of 1/2 inch diameter by microextraction method, which has been successfully used for DNA analysis.2) The PCR amplified anterior 1141 bp product from the CYP21A2 gene was digested by Kpn I, generating 309 bp, 832 bp fragments, not by Taq I, indicating its specificity.3) The frequencies of exon 1 nucleotide 138 A/C polymorphism in Korean population were 0.81, 0.91 respectively, and average heterozygosity was 0.31.4) None of four P450c21 deficient patients turned out to carry complete deletion of the CYP21A2 gene based on selective PCR amplification of the CYP21A2 gene.In conclusion, dried blood spots from Guthrie card can be sued for DNA analysis because of easy sample collection, bandling, shipment, and DNA extraction feasibility. The selective PCR amplification of the CYP 21A2 gene will pave the way for molecular characterization in P450c21 deficient patients. The exon 1 A/C polymorphism can by efficiently used for molecular diagnosis of P450c21 deficiency in informative families, though it has a drawback of handling radioactive material.
Adrenal Hyperplasia, Congenital ; Alleles ; Base Sequence ; Diagnosis ; DNA ; Exons ; Gene Frequency ; Humans ; Infant, Newborn ; Mass Screening ; Methods ; Molecular Biology ; Polymerase Chain Reaction ; Prospective Studies ; Pseudogenes ; Sensitivity and Specificity ; Sepharose ; Sequence Deletion ; Steroid 21-Hydroxylase

No abstract available.
Krukenberg Tumor*

Incidence of aseptic loosening of hip prostheses is increasing in recent years. Previous studies suggested the involvement of proteinases and cytokines in the accelerated bone lysis associated with loosening. To investigate the role of matrix metalloproteinases (MMPs) in the loosening, Gelatin/Type IV collagenases, namely, 72 KDa matrix metalloproteinase (MMP)-2 type and 92 KDa MMP-9 type were analyzed in 14 cases of the loosened endoprostheses of the hip. Zymographic and densitometric analyses revealed production of MMP-2 ancl elevated induction of MMP-9 in tissue extracts from both the interface hetween hone and implants and the capsular tissues when compared with those in synovium obtained from a patient with a t'ractured femoral neck. MMP-9 showed stronger activity than MMP-2. In the sample of a fractured femoral neck, MMP-2 was detected, but MMP-9 was not detected. In matched samples, the activity of MMP-2 and MMP-9 in the interface tissues showed stronger activity than those in the capsular tissues. There was no difference between cemented and uncemented femoral prostheses. The state of prostheses(loosening, osteolysis, and cup wear) did not influence on the activity of MMP-2 and MMP-9. Theses findings suggest a role for MMP-2 and MMP-9 type gelatinase/Type IV collagenases in the degradation of extracellular matrix of periprosthetic tissues, where they may cause weakening of the connective tissue hed and the loosening of total hip replacement endoprostheses. Consequently. we could confirm the role of MMP cascade in aseptic loosening of total hip prostheses. The further study ahout other types of MMP and the inhihitor of MMP will be needed.
Arthroplasty, Replacement, Hip ; Collagenases ; Connective Tissue ; Cytokines ; Extracellular Matrix ; Femur Neck ; Hip Prosthesis* ; Hip* ; Humans ; Incidence ; Matrix Metalloproteinases* ; Osteolysis ; Peptide Hydrolases ; Prostheses and Implants ; Synovial Membrane ; Tissue Extracts

We analyzed the clinical result of trabeculectomy with amniotic membrane transplantation which has antifibrotic and antiinflammatory action in risky glaucoma patients. Subjects were included four eyes of four patients with neovascular glaucoma, one eye of one patient with primary open angle glaucoma and four eyes of three patients with congenital glaucoma. After limbal based scleral flap was made in triangular shape with 5x4 mm sized, amniotic membrane, sized by 6x6 mm with epithelial side up, was placed on the scleral flap and then sutured to medial and lateral portion at 3 mm superior to base of scleral flap base with 10-0 nylon(2 points). The state of bleb, intraocular pressure and complications were observed at postoperative 1st, 3rd, 6th and 9th week. The IOP was reduced from 33.6+/-11.1 mmHg, to 15.5+/-6.5 mmHg at postoperative one week, 17.0+/-8.3 mmHg at three weeks, 16.9+/-8.1 mmHg at six weeks, 17.1+/-6.4 mmHg at nine weeks. 6 eyes(77.7%)had controlled IOP under 21 mmHg without antiglaucomatous agents at last follow up. Avascular blebs were observed throughout postoperative period in all subjects. Postoperative complications were shallow anterior chamber in one eye at 1st week and low intraocular pressue in one eye at 1st, 3rd week, that resolved at six weeks. Trabeculectomy with amniotic membrane transplantation on the scleral flap appeared to be effective as an augment therapy for filtering surgery in risky glaucoma.
Amnion ; Anterior Chamber ; Blister ; Filtering Surgery ; Follow-Up Studies ; Glaucoma ; Glaucoma, Neovascular ; Glaucoma, Open-Angle ; Humans ; Intraocular Pressure ; Membranes* ; Postoperative Complications ; Postoperative Period ; Trabeculectomy

The effect of YAG laser iridotomy on intraocular pressure was investigated in 24 eyes of 19 patients with chronic angle-closure glaucoma prospectively for 3 months. In most cases(75%) the intraocular presure was reduced and the mean amount of reduction was 4.33mmHg. The amount of pressure reduction has no significant correlation with the extent of peripheral anterior synechiae and visual field loss. This result shows that laser iridotomy should be considered in the management of chronic angle-closure glaucoma irrespective of the extent of peripheral anterior synechiae and visual field loss and some amount of intraocular pressure reduction may be expected after laser iridotomy.
Glaucoma, Angle-Closure* ; Humans ; Intraocular Pressure ; Lasers, Solid-State* ; Prospective Studies ; Visual Fields

No abstract available.
Cicatrix* ; Endometriosis* ; Episiotomy* ; Female

No abstract available.
Cesarean Section* ; Female ; Pregnancy

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.
Arginine ; Blood Cell Count ; Body Height ; Chemistry ; Child* ; DNA, Recombinant ; Escherichia coli ; Growth Hormone* ; Human Growth Hormone ; Humans* ; Hypoglycemia ; Insulin-Like Growth Factor I ; Korea ; Levodopa ; Plasma ; Thyroid Gland ; Urinalysis ; Vital Signs

PURPOSE: Oncostatin M(OSM) is a multifunctional cytokine that belongs to the interleukin(IL)-6 family. Although there have been a number of studies that focused on the role and mechanism of OSM in various organs and tissues, there are few reports on its effect on wound healing. The final purpose of this project is to evaluate the effect of OSM on wound healing. This pilot study was designed to investigate the effect of OSM on proliferation and matrix synthesis of human dermal fibroblasts, which are the major components of the wound healing. METHODS: Excess skin that was obtained from patients who underwent skin grafts, was used for this study. From this material, fibroblasts were isolated and cultured. The cultured fibroblasts were treated with one of four concentrations of OSM. The OSM concentrations used were 0, 50, 100, and 200ng/ml, respectively. After the OSM treatment, cell proliferation was determined by the MTT assay, collagen synthesis by the C1CP method, GAG levels by the Blyscan Dye method. The parameter levels of each group were compared. RESULTS: OSM treatment increased all the components tested in the study. In particular, cell proliferation, GAG synthesis demonstrated statistically significant increases(p<0.05 in the Mann-Whitney U-test). The highest increase in all the components was obtained at a 100ng/ml concentration of OSM. CONCLUSION: The results of the present study indicate that OSM stimulates proliferation and matrix synthesis of human dermal fibroblast and the optimal concentration for wound healing is 100ng/mL.
Cell Proliferation ; Collagen ; Fibroblasts ; Humans ; Oncostatin M ; Pilot Projects ; Skin ; Transplants ; Wound Healing

PURPOSE: We evaluated the clinical, plain radiographic, and anglographic findings of congenital vascular malformation of the soft tissue. METHODS AND MATERIALS: Retrospective analysis was performed in 36 patients. Pathological diagnosis was done in 25 patients by surgery and the others were clinically and anglographically diagnosed. On the basis of anglographic findings, we classified the lesions to three groups as arteriovenous malformation(AVM), hemangioma, and venous malformation. In pathologically proven 25 cases, we compared the anglographic diagnosis with the pathologic diagnosis. RESULTS: By anglographic classification, AVM was 13 cases, hemangioma 16 cases, and venous malformation 7 cases. The locations of the lesions were upper extremities in 14 cases, lower extremities in 20 cases, both extremities in 1 case, and back in 1 case. Clinical findings were bruit and thrill in 13 cases(12 AVMs, 1 hemangioma) and varicosities in 16 cases(11 AVMs, 3 hemangiomas and 2 venous malformations). The varicosities in AVM were pulsating nature, but not in hemangioma and venous realformation. The concordance rate of the anglographic and pathologic diagnosis was 100%(6/6) in AVM, 71%(10/14) in hemangioma and 60% (3/5) in venous malformation. CONCLUSION: We think that angiography is an essential study for accurate diagnosis and appropriate treatment of congenital vascular malformation.
Angiography ; Classification ; Diagnosis ; Extremities ; Hemangioma ; Humans ; Lower Extremity ; Retrospective Studies ; Upper Extremity ; Vascular Malformations*