No abstract available.
Seroepidemiologic Studies*

We report a case of Hunter' s syndrome with characteristic nodules on the upper back. The patient was a 7-year-old Korean boy who presented with ivory-colored papules and nodules on both sides of the scapula, pectoral regions and lateral aspects of the upper arms. These lesione are regarded as pathognomonic cutaneous markers for Hunter s syndrome. He also presented with truncal hypertrichosis, retarded growth, short neck, round face, claw like contractures of hands, multiple joint contractures, and a clear cornea. Severely elevated glycosaminoglycan levels were present in the patient s urine samples. The patient s 5-year-old brother had similar clinical features.
Animals ; Arm ; Child ; Child, Preschool ; Contracture ; Cornea ; Hand ; Hoof and Claw ; Humans ; Hypertrichosis ; Joints ; Male ; Mucopolysaccharidosis II* ; Neck ; Scapula ; Siblings

No abstract available.
beta-Thalassemia*

BACKGROUND: Stool antigen detection kits for diagnosis of infection of Helicobacter pylori have been widely used for their convenience, but are mostly imported. Since Helicobacter pylori strains show a distinctive genetic diversity, it is important to find a protein that is a common antigen among various strains and shows a strong immunogenicity for the development of a stool antigen detection kit. HP0231 protein strongly reacts with the sera of patients suffering from gastritis and peptic ulcer. Therefore, HP0231 is an excellent candidate as a target gene for this study. METHODS: Chromosomal DNA from H. pylori was isolated. HP0231 gene was amplified by PCR, cloned into pET28a(+) vector, and overexpressed using isopropyl-beta-D-thiogalactopyranoside in E. coli BL21 (DE3). HP0231 protein was purified by Ni-NTA affinity chromatography followed by electroelution after SDS-PAGE. Rabbits were immunized with the purified HP0231 protein for the production of antibodies. Rabbit anti-HP0231 antibody was partially purified and tested for the sensitivity and specificity using ELISA and Western Blot Analysis. RESULTS: The sequence of the cloned HP0231 gene was identical with the gene sequence from Genbank (AA216016). HP0231 gene was overexpressed and HP0231 protein was purified. Rabbit anti-HP0231 antibody produced after immunization with the purified HP0231 protein reacted with the purified HP0231 protein, cell extracts from cultured H. pylori, and stomach biopsy tissue from patients, but not with cell extracts from cultured E. coli used as a negative control. After 1 million fold dilution, rabbit anti-HP0231 antibody still reacted with 1 microgram of HP0231 protein. CONCLUSIONS: Rabbit anti-HP0231 antibody was produced to detect HP0231 protein of H. pylori and will be tested for the development of a stool antigen detection kit for H. pylori.
Antibodies ; Biopsy ; Blotting, Western ; Cell Extracts ; Chromatography, Affinity ; Clone Cells ; Databases, Nucleic Acid ; Diagnosis ; DNA ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Gastritis ; Genetic Variation ; Helicobacter pylori* ; Helicobacter* ; Humans ; Immunization ; Peptic Ulcer ; Polymerase Chain Reaction ; Rabbits ; Sensitivity and Specificity ; Stomach

PURPOSE: In preliminary studies, it was found that mammalian cells can be infected by recombinant baculovirus in vitro. Therefore, the potential use of recombinant baculovirus(BacG-cytonegalovirus(CMV)-P53) for the bladder gene therapy was investigated. MATERIALS AND METHODS: The recombinant Baculovirus(BV) pseudotyped was developed with the vesicular stomatitis virus(VSV) G protein. The presence of the VSV-G protein in purified BV preparations was confirmed by Western blotting analysis. The bladder cancer cells of human(HT-1376) were infected with various multiplicity of infection(MOI) of the BV, and the percentage of apoptotic cells determined by methyl thiazolyl tetrazolium(MTT) assay. RESULTS: The suppression effect of the recombinant BV with a P53 insertion in human bladder cancer cells(HT-1376) increased as the MOI of the recombinant BV increased; 100% cell survival in the group with PBS, and 81.6+/-4.3, 52.0+/-5.6 and 39.8+/-3.7% at 1, 10 and 100 MOI, respectively (p<0.05). CONCLUSIONS: Significant growth suppression was observed following infection with BacG-CMV-P53 in a human bladder cancer cell line. This observation suggests that BacG-CMV-P53 may be a potentially effective agent to prevent recurrence for P53 mutated bladder cancer. Bladder gene therapy using recombinant baculovirus could be a safe and effective treatment of bladder cancer.
Baculoviridae* ; Blotting, Western ; Cell Line* ; Cell Survival ; Genetic Therapy* ; GTP-Binding Proteins ; Humans ; Recurrence ; Urinary Bladder Neoplasms* ; Urinary Bladder* ; Vesicular Stomatitis

PURPOSE: In preliminary studies, it was found that mammalian cells can be infected by recombinant baculovirus in vitro. Therefore, the potential use of recombinant baculovirus(BacG-cytonegalovirus(CMV)-P53) for the bladder gene therapy was investigated. MATERIALS AND METHODS: The recombinant Baculovirus(BV) pseudotyped was developed with the vesicular stomatitis virus(VSV) G protein. The presence of the VSV-G protein in purified BV preparations was confirmed by Western blotting analysis. The bladder cancer cells of human(HT-1376) were infected with various multiplicity of infection(MOI) of the BV, and the percentage of apoptotic cells determined by methyl thiazolyl tetrazolium(MTT) assay. RESULTS: The suppression effect of the recombinant BV with a P53 insertion in human bladder cancer cells(HT-1376) increased as the MOI of the recombinant BV increased; 100% cell survival in the group with PBS, and 81.6+/-4.3, 52.0+/-5.6 and 39.8+/-3.7% at 1, 10 and 100 MOI, respectively (p<0.05). CONCLUSIONS: Significant growth suppression was observed following infection with BacG-CMV-P53 in a human bladder cancer cell line. This observation suggests that BacG-CMV-P53 may be a potentially effective agent to prevent recurrence for P53 mutated bladder cancer. Bladder gene therapy using recombinant baculovirus could be a safe and effective treatment of bladder cancer.
Baculoviridae* ; Blotting, Western ; Cell Line* ; Cell Survival ; Genetic Therapy* ; GTP-Binding Proteins ; Humans ; Recurrence ; Urinary Bladder Neoplasms* ; Urinary Bladder* ; Vesicular Stomatitis

PURPOSE: This study attempted to characterize the muscle derived stem cells isolated from the primary cultured skeletal muscle of the rat gastrocnemius muscle; in addition, we modified the preplate method and then compared this to the original preplate method. MATERIALS AND METHODS: The hind limbs (gastrocnemius muscles) were removed from a 3-6 week olds SD-rat and the bone was dissected away. The muscle mass was finely minced and chopped using razor blades. In an original preplate method, the cells were dissociated using a triple enzyme mixture (collagenase XI, dipase and trypsin) for 1 hour at 37degreesC. The muscle cell extract was preplated on culture flasks as described by Dr. Qu (Qu et al., 1998). The pp1-pp4 cells were referred to as the early plate (EP) cells, and the pp5-pp6 cells were referred to as the late plate (LP) cells. When we modified the preplate method, the pp1-pp2 cells were called the early plate (EP) cells and the pp3-pp4 cells were called to late plate (LP) cells. The phenotypical characteristics of EP and LP cells were compared by immunostaining and FACS. RESULTS: In the original preplate methods, the early plate (EP) cells were mixed with myogenic cells (mostly fibroblasts, <15% desmin + cells) and the LP cells were highly purified muscle derived cells that contain pure myogenic cells (>90% desmin + cells). Yet in the modified preplate method, the muscle derived stem cells were determined to be CD34 (+ or -), CD45- and desmin- cells by immunohistochemical staining and FACS. CONCLUSIONS: In original methods, the LP cells exhibited stem cell properties (CD34+, less than 30%), and they were not from a hematogeous origin (CD45-), but rather, they were from a myogenic origin (desmin+). Yet in the modified preplate method, we purified the LP cells much earlier than the original method. The LP cells displayed CD34+(more than 50%), and CD45-; thus, we isolated more primitive (desmin-) cells.
Animals ; Desmin ; Extremities ; Fibroblasts ; Muscle Cells ; Muscle, Skeletal* ; Rats* ; Stem Cells*

PURPOSE: This study attempted to characterize the muscle derived stem cells isolated from the primary cultured skeletal muscle of the rat gastrocnemius muscle; in addition, we modified the preplate method and then compared this to the original preplate method. MATERIALS AND METHODS: The hind limbs (gastrocnemius muscles) were removed from a 3-6 week olds SD-rat and the bone was dissected away. The muscle mass was finely minced and chopped using razor blades. In an original preplate method, the cells were dissociated using a triple enzyme mixture (collagenase XI, dipase and trypsin) for 1 hour at 37degreesC. The muscle cell extract was preplated on culture flasks as described by Dr. Qu (Qu et al., 1998). The pp1-pp4 cells were referred to as the early plate (EP) cells, and the pp5-pp6 cells were referred to as the late plate (LP) cells. When we modified the preplate method, the pp1-pp2 cells were called the early plate (EP) cells and the pp3-pp4 cells were called to late plate (LP) cells. The phenotypical characteristics of EP and LP cells were compared by immunostaining and FACS. RESULTS: In the original preplate methods, the early plate (EP) cells were mixed with myogenic cells (mostly fibroblasts, <15% desmin + cells) and the LP cells were highly purified muscle derived cells that contain pure myogenic cells (>90% desmin + cells). Yet in the modified preplate method, the muscle derived stem cells were determined to be CD34 (+ or -), CD45- and desmin- cells by immunohistochemical staining and FACS. CONCLUSIONS: In original methods, the LP cells exhibited stem cell properties (CD34+, less than 30%), and they were not from a hematogeous origin (CD45-), but rather, they were from a myogenic origin (desmin+). Yet in the modified preplate method, we purified the LP cells much earlier than the original method. The LP cells displayed CD34+(more than 50%), and CD45-; thus, we isolated more primitive (desmin-) cells.
Animals ; Desmin ; Extremities ; Fibroblasts ; Muscle Cells ; Muscle, Skeletal* ; Rats* ; Stem Cells*

BACKGROUND: In recent years, the incidence of extended-spectrum beta-lactamase (ESBL) producing Klebsiella has been steadily increased, and the newer species K. planticola and K terrigena, formerly regarded as nonpathogen, have been reported with astonishing frequency from human infectious processes by some investigators. The aim of this study is to elucidate the isolation rate and antimicrobial susceptibility of recent clinical Klebsiella isolates. METHOD: For the clinical Klebsiella isolates during the period of June 1999 to May 2000, isolation frequency of Klebsiella species by specimen, departments, age, and sex were analyzed. And antimicrobial susceptibilities were also analyzed. RESULT: Isolation rate of Klebsiella in order of decreasing frequency were K. pneumoniae (74:7%), K. oxytoca (12.1%), K. ozaenae(1.7%), K. planticola(1.0%), K. terngena(0.9%), and K, ornithinolytica (0.7%), respectively. K. rhinoscleromatis was not isolated. Compared with outpatients, increase of resistance rates of inpatients's Klebsiella isolates were 10% in ciprofloxacin, 15% in cefoperazone/sulbactam, and the others were ranged from 24% to 31%. Isolation rate of ESBL producing K. pneumoniae by double disk (DD) synergy test was 41%, and detection rates by antimicrobial agents were as follows: cefotaxime (95%), aztreonam (58%), and ceftriaxone (37%). Antimicrobial susceptibility rate with the exception of ampicillin and imipenem decreased from the range of 81%-96% on admission day to 29-62% after one week on admission. CONCLUSION: The isolation rates of K. planticola and K. terrigena were less than 1%. The proportion of ESBL producing K. pneumoniae was 41 %. And the vast majority of multidrug resistant Klebsiella including ESBL producing strains are acquired by hospitalization.
Ampicillin ; Anti-Infective Agents ; Aztreonam ; beta-Lactamases ; Cefotaxime ; Ceftriaxone ; Ciprofloxacin ; Epidemiology* ; Hospitalization ; Humans ; Imipenem ; Incidence ; Klebsiella pneumoniae ; Klebsiella* ; Outpatients ; Pneumonia ; Research Personnel