Human gingival fibroblasts have proven to useful as a species specific cell culture system in various system on periodontal disease and regeneration. However, their use is limited, since they are hard to obtain and lifespan is short due to replicative senescence. To overcome these disadvantages, we transfected primary human gingival fibroblasts by the E6 and E7 genes of the Human papilloma virus(HPV) 16. The full length of HPV 16 E6 and E7 was cloned from the pBR322 into BamH1 and Sal I of a pBabe vector including hygromycin B resistance. Before pBabeE6/E7 plasmid transfection, peak 8 GFP including G418 resistance was transfected into primary GF to check the transfection efficency. PBabe E6/E7 plasmid was transfected using Lipofectamine plus following manufacter's instruction into primary normal human gingival fibroblasts in 60mm dishes with FBS free DMEM. After 2 days of transfection, the cells were treated with hygromycin for 2 weeks until the transfected control cells died. The resulting hygromycin resistant colonies were pooled, and clonned, and sucessful transfection was established for immortalized gingival fibroblast cell lines. Immoralized GF cells showed stellate shape, that is similar to that of orange grains, and more rapid growth and higher proliferation than that of primary gingival fibroblasts. This cell lines overcame crisis and could be cultured over 30 subcultured, could be use for three dimentional culture, epithelial-mesenchymal interaction study.
Cell Aging ; Cell Culture Techniques ; Cell Line* ; Edible Grain ; Citrus sinensis ; Clone Cells ; Fibroblasts* ; Human papillomavirus 16 ; Humans* ; Hygromycin B ; Papilloma ; Periodontal Diseases ; Plasmids ; Regeneration ; Transfection

The ultimate goal of periodontal therapy is the regeneration of periodontal tissue and the repair of function. For more than a decade there have been many efforts to develop ma- terials and methods of treatment to promote periodontal tissue regeneration. Recently many efforts are concentrated on the regeneration potential of material used in traditional medicine. Safflower(Carthamus tinctorius L.) seed extract(SSE) have long clinically used in Korea to promote bone formation and prevent osteoporosis. The purpose of this study was to examine the effects of SSE on bone formation in human osteoblastic cell line. Human fetal osteoblastic cell line(hFOB 1.19) was cultured with DMEM and SSE(1microgram/ml, 10microgram/ml, 100microgram/ml, 1mg/ml) at 34degrees C with 5% CO2 in 100% humidity. The proliferation, differentiation of the cell was evaluated by several experiments. Cell proliferation was significantly increased at 10microgram/ml, 100microgram/ml, 1mg/ml of SSE after 3 and 7 days incubation(p<0.05). Cell spreading assay was significantly increased at 100microgram/ml of SSE after 3 days and 1microgram/ml, 10microgram/ml, 100microgram/ml, 1mg/ml of SSE after 7 days(p<0.05). Alkaline Phosphatase(ALP) level was significantly increased in 10microgram/ml, 100microgram/ml of SSE(p<0.05). Collagen synthesis was significantly increased at 10microgram/ml, 100microgram/ml, 1mg/ml of SSE(p<0.05). A quantified calcium accumulation was significantly increased at 10microgram/ml, 100microgram/ml of SSE(p<0.05). ALP and osteocalcin mRNA was expressed in 100microgram/ml of SSE by RT-PCR. These results indicate that SSE are capable of increasing osteoblasts mineralization and may play an important role in bone formation.
Calcium ; Carthamus tinctorius* ; Cell Line ; Cell Proliferation ; Collagen ; Humans ; Humidity ; Korea ; Medicine, Traditional ; Osteoblasts ; Osteocalcin ; Osteogenesis* ; Osteoporosis ; Regeneration ; RNA, Messenger

BACKGROUND AND OBJECTIVES: Maxillary sinus mucoceles are usually developed long after Caldwell-Luc's operation, which are known as postoperative cheek cysts and common in Korea. However, primary mucoceles of the maxillary sinus are relatively rare compared to frontal or ethmoid sinus mucoceles. This study aims to describe the clinical manifestation of the primary maxillary sinus mucoceles and to determine the long-term efficacy of the surgical treatment. MATERIALS AND METHOD: Seven cases, comprised of 2 women and 5 men with an age range of 23 to 72 years, finally diagnosed as primary maxillary sinus muco (pyo)celes between March 1995 and July 2004 were studied. The presenting signs and symptoms, radiological findings, and surgical management were retrospectively reviewed. RESULTS: All patients complained of nasal obstruction, while only one patient had cheek pressure and pain. In the endoscopic nasal examination, the medial wall of maxillary sinus was bulging with prolapsed middle meatal mucosa in 6 cases. Five patients were treated with endonasal endoscopic sinus surgery and two with sublabial approach. Intraoperative cultures grew organisms in four patients. In all patients, the middle meatal antrostomy openings were patent without recurrence at the latest follow-up ranging from 11 to 47 months. CONCLUSION: A primary maxillary sinus mucocele appears as a bulging medial wall with unilateral nasal obstruction. Endoscopic sinus surgery is a reliable therapeutic measure with lesser morbidity and favorable long-term outcome.
Cheek ; Endoscopy ; Ethmoid Sinus ; Female ; Follow-Up Studies ; Humans ; Korea ; Male ; Maxillary Sinus* ; Mucocele* ; Mucous Membrane ; Nasal Obstruction ; Recurrence ; Retrospective Studies

STATEMENT OF PROBLEM: There have been few studies about the marginal fit of Auro Galvano Crowns. PURPOSE: The purpose of this study was to compare the marginal fit of the anterior single restorations. MATERIAL AND METHODS: The in vitro marginal discrepancies of metal-ceramic, Auro Galvano Crown and coping were evaluated and compared. The Auro Gavano Crowns were made from one extracted maxillary central incisor prepared by milling machine. 30crowns per each system were fabricated. Measurements of a crown were recorded at 50 points that were randomly selected for marginal gap evaluation. Parametric statistical analysis was performed for the results. RESULTS: Mean marginal gap dimensions and standard deviations at the marginal opening for the anterior single crowns were 74+/-21micrometer for the control (metal-ceramic restoration), 45+/-11 micrometer for Auro Galvano Crown coping, and 51+/-9 micrometer for the Auro Galvano Crown. CONCLUSIONS: Auro Galvano Crown showed significantly smaller (P< .05) marginal gap than the control. Ceramic application did not significantly affected the marginal fit of Auro Galvano Crown. (P> .05)
Ceramics ; Crowns* ; Incisor

This study is to evaluate clinical and arthroscopic second-look results of arthroscopic repairs of posterior root tears of medial meniscus which may cause loss of circumferential hoop tension and extrusion of meniscus. From October 2006 to May 2009, fifty-eight patients (59 knees) underwent arthroscopic pull-out repairs. Clinical results were evaluated using Hospital for Special Surgery (HSS) score and International Knee Documentation Committee (IKDC) score for 12-month follow-up. Second-look arthroscopy was done to evaluate meniscal healing in 21 cases. Magnetic resonance imaging (MRI) was performed to assess status of repaired meniscus and tibial tunnel position in 9 patients. Average preoperative HSS score and IKDC score of 59 cases were 69.5 and 36.0, respectively. Average postoperative HSS score and IKDC score of 59 cases had been changed into 90.3 (p<0.001) and 66.8 (p<0.001), respectively. Second-look arthroscopies revealed complete or incomplete healing except one case. Two patients showed increased one grade according to the Kellgren-Lawrence radiologic classification system and others showed no change. Of 9 patients who performed MRI, six patients showed complete healing. The average position of tibial tunnel was 4.8 mm anterior and 5.7 mm medial to center of posterior cruciate ligament. Arthroscopic pull-out repair technique using transtibial tunnel seems to be simple and effective procedure for posterior root tear of medial meniscus. Further evaluation of arthroscopic repair of posterior root tear of medial meniscus should be needed to prove the effectiveness on the prevention of osteoarthritis of knee.
Arthroscopy ; Follow-Up Studies ; Humans ; Knee ; Magnetic Resonance Imaging ; Menisci, Tibial ; Osteoarthritis, Knee ; Posterior Cruciate Ligament

Trichloroethylene is commonly used as an industrial solvent and degreasing agent. The clinical features of acute and chronic intoxication with trichloroethylene are well-known and have been described in many reports, but hypersensitivity syndrome caused by trichloroethylene is rarely encountered. For managing patients with trichloroethylene hypersensitivity syndrome, avoiding trichloroethylene and initiating glucocorticoid have been generally accepted. Generally, glucocorticoid had been tapered as trichloroethylene hypersensitivity syndrome had ameliorated. However, we encountered a typical case of trichloroethylene hypersensitivity syndrome refractory to high dose glucocorticoid treatment. A 54-year-old Korean man developed jaundice, fever, red sore eyes, and generalized erythematous maculopapular rashes. A detailed history revealed occupational exposure to trichloroethylene. After starting intravenous methylprednisolone, his clinical condition improved remarkably, but we could not reduce prednisolone because his liver enzyme and total bilirubin began to rise within 2 days after reducing prednisolone under 60 mg/day. We recommended an extended admission for complete recovery, but the patient decided to leave the hospital against medical advice. The patient visited the emergency department due to pneumonia and developed asystole, which did not respond to resuscitation.
Dermatitis, Occupational/*etiology ; Fatal Outcome ; Humans ; Hypersensitivity/*etiology ; Male ; Middle Aged ; Occupational Exposure/*adverse effects ; Solvents/*toxicity ; Trichloroethylene/*toxicity

PURPOSE: To review clinical and radiological results after open reduction and internal fixation with T plate for unstable distal clavicle fractures. MATERIALS AND METHODS: From July. 1999 to December 2002, nine patients with distal clavicle Neer type II fractures were treated by open reduction and internal fixation with T plate. The bony union was confirmed by plain radiography. The clinical results were analyzed according to the classification by Kona et al. RESULTS: Average time to fracture union was 8 weeks in all cases. The functional results were as follows: excellent in 7 cases and good in 2 cases. Screw loosening occurred in one case, but bony union was achieved. CONCLUSION: We recommend T plate fixation as another treatment method for unstable distal clavicle fractures.
Classification ; Clavicle* ; Humans ; Radiography

BACKGROUND: The development of new drugs or alternative therapies effective against methicillin-resistant Staphylococcus aureus (MRSA) is of great importance, and various natural anti-MRSA products are good candidates for combination therapies. We evaluated the antibacterial activities of a Phellinus baumii ethyl acetate extract (PBEAE) and its synergistic effects with beta-lactams against MRSA. METHODS: The broth microdilution method was used to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the PBEAE. The PBEAE synergistic effects were determined by evaluating the MICs of anti-staphylococcal antibiotic mixtures, with or without PBEAE. Anti-MRSA synergistic bactericidal effects of the PBEAE and beta-lactams were assessed by time-killing assay. An ELISA was used to determine the effect of the PBEAE on penicillin binding protein (PBP)2a production. RESULTS: The MICs and MBCs of PBEAE against MRSA were 256-512 and 1,024-2,048 microg/mL, respectively. The PBEAE significantly reduced MICs of all beta-lactams tested, including oxacillin, cefazolin, cefepime, and penicillin. However, the PBEAE had little or no effect on the activity of non-beta-lactams. Time-killing assays showed that the synergistic effects of two beta-lactams (oxacillin and cefazolin) with the PBEAE were bactericidal in nature (Deltalog10 colony forming unit/mL at 24 hr: 2.34-2.87 and 2.10-3.04, respectively). The PBEAE induced a dose-dependent decrease in PBP2a production by MRSA, suggesting that the inhibition of PBP2a production was a major synergistic mechanism between the beta-lactams and the PBEAE. CONCLUSIONS: PBEAE can enhance the efficacy of beta-lactams for combined therapy in patients infected with MRSA.
Acetates/chemistry ; Agaricales/*chemistry/metabolism ; Anti-Infective Agents/chemistry/*pharmacology ; Drug Synergism ; Enzyme-Linked Immunosorbent Assay ; Methicillin-Resistant Staphylococcus aureus/*drug effects/metabolism ; Microbial Sensitivity Tests ; Penicillin-Binding Proteins/analysis/metabolism ; Plant Extracts/chemistry/*pharmacology ; beta-Lactams/*pharmacology

BACKGROUND: Initial symptoms for esophageal perforation have not been clarified, but when there is no early diagnosis and proper treatment to follow immediately after the diagnosis, it is fatal for the patients. Therefore, this study attempted to discover the factors that influence the prognosis of esophageal perforation to contribute to the improvement of the treatment result. MATERIAL AND METHOD: The subjects of this study are 32 patients who came to the hospital with esophageal perforation from October, 1984 to June, 2000. This study examined the items for clinical observation such as patients' sex, age, cause of the perforation, perforation site, the time spent until the beginning of the treatment, symptoms caused by the perforation and its complication, and treatment methods. This study tried to find out the relationship between the survival of patients and each item. RESULT: There were 24 male and 8 female patients and their mean age was 49.7 +/- 16.4. For the causes of perforation, there were 14 cases(43%) of iatrogenic perforation, which ranked first, caused by the medical instrument operation and surgical damage. As for the perforation sites, thoracic esophagus was the most common site(26 cases of 81.2%) and chest pain was the most frequent symptom. The complication caused by esophageal perforation showed the highest cases in the order of mediastinitis, empyema, sepsis and peritonitis. After the treatment, there were 23 cases of survival and 9 cases of mortality. The total mortality rate was 28.1% and the main causes of mortality were sepsis and acute respiratory distress syndrome(ARDS). As for the treatment, 8 cases(25.0%) treated the perforation successfully using conservative treatment only. As for the surgical treatment, there were 5 cases(15.6%) of cervical drainage, 7 cases (21.8%) of primary repair and 12 cases(37.5%) of esophageal reconstruction after performing an exclusion-diversion. There were 18 cases(56.2%) of complete treatment of esophageal perforation at its initial treatment and in 14 cases(43.8%) of treatment failure at its initial treatment, patients were completely cured in the next treatment stage or died during the treatment. The cases of perforation in thoracic esophagus, complication into severe mediastinitis or sepsis and the cases of failure at initial treatment showed a statistically significant mortality rate (p<0.05). CONCLUSION: Based on the above results, it is thought that a proper choice for initial treatment choice depending on the perforation site and the prevention of serious complication such as mediastinitis or sepsis can shorten the treatment period for the patients with esophageal perforation and improve the convalescence.
Chest Pain ; Convalescence ; Diagnosis ; Drainage ; Early Diagnosis ; Empyema ; Esophageal Perforation* ; Esophagus ; Female ; Humans ; Male ; Mediastinitis ; Mortality ; Peritonitis ; Prognosis ; Sepsis ; Treatment Failure

OBJECTIVES: Pregnanolone is a potent positive modulator of the gamma-aminobutyric acid(GABA) response that enhances the binding of [3H]flunitrazepam to the GABA A receptor. Recently, it was reported that chronic treatment with pregnanolone uncouples allosteric interactions between steroid and benzodiazepine recognition sites. The present study was designed to assess the effect of repeated stress on the modulation of neuroactive steroids on the GABA A receptor. METHODS: The effect of steroids on the ligands binding to GABA A receptor was investigated using cerebral cortices of unstressed and repeatedly immobilized rats. Male Sprague-Dawley rats, weighing 200-250g were forced to suffer an immobilization stress for 2 hours. RESULTS: Pregnanolone enhanced the binding of [3H]flunitrazepam to GABA A receptor in both of unstressed and repeatedly stressed rats. However, repeatedly stressed rats showed significantly higher values in EC50 and lower values in E max of enhancement binding of [3H]flunitrazepam than those of unstressed rats. CONCLUSIONS: From these findings, it can be concluded that repeated stress reduced the positive modulation of neuroactive steroid on the GABA A-receptor complex.
Animals ; Benzodiazepines ; Cerebral Cortex ; gamma-Aminobutyric Acid ; Humans ; Immobilization ; Ligands ; Male ; Pregnanolone ; Rats* ; Rats, Sprague-Dawley ; Receptors, GABA-A ; Steroids*