In view of the potential of replicase protein as a diagnostic reagent for human caliciviruses (HuCVs), we have cloned and over-expressed this gene from the Snow Mountain-like Korean strains in Escherichia coli as a fusion protein with glutathione S-transferase (GST, and described the preliminary antigenic characterization of the recombinant products. Each 470bp fragment corresponding to highly conserved region of RNA-dependent RNA polymerase was generated by RT-PCR from stools of two diarrheal children, cloned in pMOSBlue T-vector, and subcloned between the EcoRI and SalI restriction sites of pGEX-47-3, a GST gene fusion vector, yielding pGCVpol. This construct expressed a Snow Mountain-like HuCV replicate under the control of the IPTG-inducible pac promoter. An extract prepared by sonication of the E. coli cell inclusion bodies bearing pGCVpol products was purified and analyzed by SDS-PAGE. After Coomassie blue staining, it was shown that the recombinant replicase migrated on the gels with an approximate molecular mass of 46.5 kDa, that was subsequently cleaved into a 26 kDa GST fragment and a 20.5 kDa replicase protein upon digestion with thrombin protease. The replicase was recognized on immunoblotting with the sera from symptomatic children with the HuCV-associated diarrhea but not by asymptomatic sera from adults. The results presented the first biological activity of individually expressed HuCV replicase subunit and provided important reagents for diagnosis of HuCV infection.
Adult ; Child ; Clone Cells ; Diagnosis ; Diarrhea ; Digestion ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Gels ; Gene Fusion ; Glutathione Transferase ; Humans ; Immunoblotting ; Inclusion Bodies ; Indicators and Reagents ; RNA Replicase ; Snow* ; Sonication ; Staphylococcal Protein A ; Thrombin

Porencephaly is relatively rare condition defined by an defect or a defect or cavity in the cerebrum owing to a developmental malformation or to a destructive lesion. Fory-five porencephaly patients diagnosed by Brain CT were clinically analyzed and the following results were obtained. 1) By the age group presenting initial symptoms, the peak incidence was from 1 month to below 3 years old. 2) In initial symptoms, seizure, spastic weakness, headache were showed in order of frequency. But 7 cases (15.5%) were asymptomatic. 3) The latency of diagnosis after presenting initial symptoms from the symptom onset time to 10 years. 4) The subsequent symptoms were as follows: spastic weakness, speech disturbance, gait disturbance, mental retardation, sensory loss and seizure showed independently or combined. 5) As etiologic factor, 21 cases (46.7%) were congenital, 16 cases (35.5%) were post-traumatic or post-operative and 8 cases (17.8%) were perinatal. 6) The prognosis was seen various from mild to severe. Out of 45 cases, 29 cases (64.4%) were no complications. But the prognosis in patients with post-traumatic or postoperative etiological factors was poor. With the advent of brain CT and the resultant capability of detecting structural defect and cerebral lesions responsible for epilepsy or focal neurologic signs, porencephaly was seen to be readily recongizable by CT examination. Since porencephaly is a significant contributor to the spectrum of CNS lesion and benign condition, ist recognition is important in determining prognosis and therapy.
Brain ; Cerebrum ; Child, Preschool ; Diagnosis ; Epilepsy ; Gait ; Headache ; Humans ; Incidence ; Intellectual Disability ; Muscle Spasticity ; Neurologic Manifestations ; Prognosis ; Seizures

No abstract available.
C-Reactive Protein* ; Chorioamnionitis* ; Female ; Humans ; Membranes* ; Pregnancy ; Rupture*

Peri-vater divertieula may interfere with the normal flow of biliary and pancreatic secretion, thus leading to pancreatobiliary diseasa and the very presence of juxtapapillary diverticulum makes cannulation technically difficult and contributes to the low success rate. In this present series, the relationship between duodenal diverticula and age, sex, and biliary-pancreatic pathology was stud.ied in 1400 patients examined with ERCP. The results are as follows. 1) Duodenal diverticula was demonstrated in 73 of 1400 patients(5, 2%). 2) The incidence of duodenal divertieular increased with age and they were more frequent in men. 3) The cannulation of one or both duct system was suecessful in 69 of the 73 patients, the success rate was 94. 5%. 4) The biliary pathology was found in 41 of the 73 patients and the pancreatic pathology in 6 of the 73 patients. The rnost common finding was biliary calculi which were demonstrated in 35 patients(47. 9%).
Catheterization ; Cholangiopancreatography, Endoscopic Retrograde* ; Diagnosis* ; Diverticulum* ; Gallstones ; Humans ; Incidence ; Male ; Pathology

Aneurysm of the superficial temporal artery is rare. We reviewed seven cases of aneurysm of the superficial temporal artery, which were confirmed surgically and angiographically. The results were as follows: The most common site of aneurysm was left superficial temporal artery, major feeding artery was anterior branch of superficial temporal artery, mean diameter was 7.8mm×12.6mm, and all aneurysms showed lobulated margin with intraluminal filling defects.
Aneurysm* ; Arteries ; Temporal Arteries*

No abstract available.
Animals ; Rats* ; Urinary Bladder*

Hypospadias can be repaired in the majority cases in 1-stage with a high success rate. We reviewed clinical experience of 50 patients who underwent primary hypospadias repair by 1 surgeon using 1-stage repairs except 3 cases in a 30-month period. The results were 1). The half of patients were younger than 5 years (48%). 2). A total of 34 cases (68.0%) had an associated ventral curvature. The dorsal tunica albuginea plication to correct intrinsic curvature was done in 21 cases (61.8%) and dividing the urethral plate in only 3 cases (8.8%). 3). Associated anomalies were cryptorchidism in 7, hydrocele in 3 and severe penoscrotal transposition in 8 cases. 4). 19 cases of anterior hypospadias were repaired by meatoplasty using Heineke-Mikulicz tissue rearrangement (9 cases), MAGPI (3 cases), pyramid procedure (4 cases), Mathieu urethroplasty (2 cases) and onlay island flap (1 case). Of the 31 cases of middle and posterior hypospadias onlay island flap repair was performed in 24 cases (77.4%). 5). The success rates in anterior, middle and posterior hypospadias were 94.7, 64.7 and 42.9%, respectively. The overall success rate was 70%. 6). The most commonly used procedure was onlay island flap urethroplasty. Only 14 of 25 cases (56.0%) were successful with this method but the last 8 cases were repaired with no complications. 7). The most common complications were urethrocutaneous fistula (11 cases) and meatal retraction (3 cases). The complications were managed by rotational advancement flap (11 cases), meatal based flap (2 cases) and urethral advancement (1 case) and the success rate was 73.3%. In conclusion, to achieve more satisfactory results the choice of operation depends on the configuration of either glans or meatus, presence or absence of chordee and status of ventral penile skin in anterior hypospadias. Although the 2-stage repair remains a safe and reliable alternatives for severe hypospadias 1-stage repair was more useful except severe hypospadias combined with penoscrotal transposition."
Cryptorchidism ; Female ; Fistula ; Humans ; Hypospadias* ; Inlays ; Male ; Skin

BACKGROUND: Diagnosis of paucibacillary leprosy is difficult owing to lack of sensitive diagnostic tools. Recently, several investigators have studied the use of polymerase chain reaction(PCR) to detect Mycabacterium leprae. Using nested-PCR the sensitivity and specif city of DNA amplification is considerably improved. OBJECTIVE: The purpose on investigation is to assess the efficacy if nested-PCR which is applied to formalin-fixed, paraffin-embedded biopsies material of patients with 1 prosy. METHODS: Biopsy samples were taken from patients with lepremc tous(11 patients) and tuberculoid (10 patients) leprosy, fixed in formalin, and embedded in parafin. The DNA from samples was extracted and amplified through 25 cycles by using the outside pairs of primer(L and L). The second amplification was allowed thproceed through 15 cycles using insice gairs of primer(L and L4). RESULTS: All twenty one samples showed 347-base-pair products. To confirm that the 347-bp product did correspond to the expected portion of the M. leprae groE gene, the amplified product was digested with Pst I. Pst I dipestion yielded 254-and 93-bp fragmerts, as predicted from the sequence of the M. leprae gene. The senilivity was that a single organism was idntified by nested-PCR. CONCLUSION: The nested-PCR is sensitive, specific, and simple diagiostic tool for leprosy.
Biopsy ; Diagnosis ; DNA ; Formaldehyde ; Humans* ; Keratinocytes* ; Leprosy ; Leprosy, Paucibacillary ; Melanocytes* ; Panax* ; Research Personnel ; Saponins* ; Water*

The cutaneous synthesis of vitamin D in response to ultraviolet radiation exposure is the most important factor in maintaining vitamin D balance in Man. The skin is not only the site of vitamin D synthesis, but also a target organ for calcitriol(1, 25-dihydroxy-vitamin D) which is naturally occuriag, hormonally active form of vitamin E. It is now known that calcitriol inhibits the proliferation of epidermal cells and induces her differentiation. In this study, epidermal keratinocytes and melanocytes were isilated from the neonatal foreskin, and were culturod using a MCDB 153 and modified TIC media, respectively. And then various concentratioris of calcipotriol(MC 903), a synthetic aralogue of calcitriol, were added to each culture. The effects of calcipotriol on the growth of human keratinocytes and melanocytes were investigated. The results were as follows : 1. The addition of calopotriol to human keratinocyte and melalocyte cultures inhibited their proliferation in a dosdependent manner. 2. Calcipotriol had no effects on the melanization process of the melanocyte. 3. Calcipotriol was found to inhibit the proliferation, however it induced the terminal differentiation of cultured keratinocytes, as judged by morphologicai changes. The decreased density of kerationcytes, The formation of cornified cells, and the cellular destruction in a concentration of 10 M of calcipotriol were observed. 4. By using the light atid the electron microscope, we observed that the epidermal thickness was decreased and terminal differentiation was facilitateir. Living Skin Equivalent (LSE) according to the increasing concentration of calcipotriol. A]i)parent cytotoxic effects were observed in 10 M, 10 M of calcipotriol. In summary, the above results indicate that the addition of calcipotriol to the in vitro culture system of human keratinocyte and melanocyte induces the biologic process of terminal differentiation of keratinocytes and inhibits proliferation of keratinoytes and melanocytes in a dose-dependent manner.
Calcitriol ; Cholecalciferol* ; Foreskin ; Humans* ; Keratinocytes* ; Melanocytes* ; Skin ; Tics ; Vitamin D ; Vitamin E ; Vitamins*

Luteal phase defect (LPD) is an ovulatory disorder of considerable clinical importance that leads to delayed endometrial maturation and inadequate endometrium for blastocyst implantation. This disorder is implicated in infertility and recurrent spontaneous abortion. We analyzed the endometrial histological maturation, and the estrogen receptor(ER) and progesterone receptor(PR) status in patients with unexplained delayed ovulation, and in patients with ovulation induction by clomiphen citrate. The purpose of this study was to determine whether the length of the follicular phase influences the endometrial histological maturation and the hormonal receptor status, and to know the effect of clomiphen citrate on the endometrium. In the delayed ovulation group, the endometrium was characterized by an irregular outpouching and dilated lumina of the glands, a decreased secretory activity and predecidualization, and a decreased number of granulocytes in the stroma. In the clomiphen citrate-ovulation induction group, glandular proliferation was markedly diminished with poorly convoluted and narrow glands, secretory activity was decreased, stroma was undecidualized, and there was an absence of granulocytes. ER expression was increased in the glandular epithelium in the delayed ovulation group, and both ER and PR expressions were markedly decreased in the glandular epithelium and stromal cells in the ovulation induction group. Endometrial maturation and differentiation may be diminished by increased estrogenic and relatively deficient progesterone effects in patients with delayed ovulation. The hormonal receptor status might also be influenced by hormonal changes. Clomiphen citrate successfully induced ovulation in patients with delayed ovulation, however, endometrial proliferation and maturation were markedly suppressed. This might be related to the lower pregnancy rate and higher abortion rate in patients with clomiphen citrate induced ovulation.
Pregnancy ; Female ; Humans