BACKGROUND: Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on it9 potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to host In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-α CDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-α gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as we]1 as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-α gene transfected cancer cells. METHOD: We transfected TNF-α c-DNA to WEHI l64, a murine fibrosarcoma cell line, using retroviral vector (pLT12SN(TNF)) and confirm the expression of TNF with PCRf ELISA, MTT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI 164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide(translation inhibitor). RESULTS: WEHI 164 which was sensitive to TNF became resistant to TNF after being trsnsfected with TNF-α gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. CONCLUSION: The acquired resistance to TNF after TNF-α gene transfection may be associated with synthesis of some protective proteins.
Biology ; Cell Line ; Cycloheximide ; Dactinomycin ; DNA, Complementary ; Enzyme-Linked Immunosorbent Assay ; Fibrosarcoma ; Genes, Neoplasm ; Necrosis ; Transfection ; Tumor Necrosis Factor-alpha* ; Zidovudine*

The aim of this study was to evaluate the differences in setting times based on the methods for dental root canal sealers and calcium silicate cement used in root-end filling. Five kinds of dental root canal sealers and four kinds of calcium silicate cement for root-end filling were selected for the experiments. All materials were mixed according to the manufacturers’ instructions and stored at 37 ℃ with a relative humidity of 95%. Setting time was measured using a 1/4 pound Gillmore needle and a 1 pound Gillmore needle to determine the time until indentation was no longer visible or the time until 2 mm penetration was no longer possible. The determination of indentation was based on the absence of visible impressions on the material surface when Gillmore needle was placed vertically. When comparing indentation time and penetration time using same type of Gillmore needle, only ProRoot MTA using 1 pound Gillmore needle showed significant difference between measuring methods (P<0.05) while there are no differences in measuring methods in other materials (P>0.05). By this study, we could expect to measure a setting time relatively similar to real clinical conditions through indentation method.

No abstract available.
Erythromycin*

BACKGROUND: Tissue inhibitor of metalloproteinase is a natural inhibitor that counteracts proteolytic enzymes essential to the invasion of cancer cell. Whether or not TIMP-2 gene transfer via adenovirus could inhibit the invasion of lung cancer cell in vitro was evaluated for the future purpose of gene therapy against lung cancer. METHODS: Recombinant adenvirus-TIMP-2(Ad-TIMP-2) was generated by homologeous recombination after pACCMV-TIMP-2 and pJM17 were cotransfected into 293 cell by standard calcium phosphate coprecipitate mathod. Calu-6, one of the most invasive lung cancer cells, was transduced with Ad-TIMP-2 or Ad-β-gal. An-chorage-independent growth and invasiveness were assessed by soft agar clonogenicity assay and invasion assay using two-chamber, well divided by matrigel. RESULTS: Ad-TIMP-2 transduced calu-6 cells produced bilolgically active TIMP-2 more than 50 times more than parental calu-6. TIMP-2 gene transfer did not suppress the in vitro tumorgenicity. However, two chamber well assay revealed that Ad-TIMP-2 transduction reduced the invasiveness of calu-6 efficiently (12% compared with parental cell) even at low 10moi. CONCLUSION: Even though TIMP-2 gene transfer did not inhibit in vitr tumorigenicity, it did inhibit invasion of lung cancer cell in vitro. The inhibition of invasion by Ad-TIMP-2 may be a useful strategy for the treatment of lung cancer.
Adenoviridae* ; Agar ; Calcium ; Genetic Therapy ; Humans ; Lung Neoplasms* ; Lung* ; Parents ; Peptide Hydrolases ; Recombination, Genetic ; Tissue Inhibitor of Metalloproteinase-2*

In patients with pupil-involving isolated oculomotor nerve palsy (IOP), compressive lesions, meningitis, and midbrain infarction can usually be excluded if the responsible lesions are not verified on a brain MRI, catheter angiography, and cerebrospinal fluid examination. However, we experienced a 11-year-old boy with pupil-involving IOP whose initial investigations had been normal and whose symptoms had been stationary over 3 months but was eventually confirmed to have germinoma. This case suggests that a patient with IOP without improvement for a prolonged period needs re-evaluation regardless of normal initial investigations. (J Korean Neurol Assoc 19(4):423~426, 2001)
Angiography ; Brain ; Catheters ; Cerebrospinal Fluid ; Child ; Germinoma* ; Humans ; Infarction ; Magnetic Resonance Imaging ; Male ; Meningitis ; Mesencephalon ; Oculomotor Nerve Diseases* ; Oculomotor Nerve*

OBJECTIVES: Insertion of a silicone stent during endoscopic dacryocystorhinostomy (DCR) is the most common procedure to prevent rhinostomy closure. It has been claimed that silicone intubation improves the surgical outcomes of endoscopic DCR. However, many reports have documented an equally high success rate for surgery without silicone intubation. Accordingly, we conducted a systematic review and meta-analysis to clarify the outcomes of endoscopic DCR with and without silicone intubation and determine whether silicone intubation is actually beneficial for patients. METHODS: PubMed, Embase, and Cochrane Library databases were searched to identify relevant controlled trials evaluating endoscopic DCR with and without silicone intubation. The search was restricted to English articles published between January 2007 and December 2016. Relevant articles were reviewed to obtain information pertaining to interventions and outcomes. We also performed a meta-analysis of the relevant literature. RESULTS: In total, 1,216 patients included in 12 randomized controlled trials were pooled. A total of 1,239 endoscopic DCR procedures were performed, and silicone stents were used in 533 procedures. The overall success rate for endoscopic DCR was 91.9% (1,139/1,239), while the success rates with and without silicone intubation were 92.9% (495/533) and 91.2% (644/706), respectively. There was no statistically significant heterogeneity among the included studies. A meta-analysis using a fixed-effects models showed no significant difference in the success rate between endoscopic DCR with silicone intubation and that without silicone intubation (OR, 1.38; 95% CI, 0.89 to 2.12; P=0.148; z=1.45). Furthermore, there were no significant differences with regard to surgical complications such as synechia, granulation, and postoperative bleeding. CONCLUSION: The findings of our meta-analysis suggest that the success rate and postoperative complication rate for endoscopic DCR is not influenced by the use of silicone intubation during the procedure.
Dacryocystorhinostomy* ; Endoscopy ; Hemorrhage ; Humans ; Intubation* ; Population Characteristics ; Postoperative Complications ; Silicon* ; Silicones* ; Stents

Segniliparus species is a novel genus that is reported to be the new emerging respiratory pathogens. Here, we report a very rare case of S. rugosus pulmonary infection in an immunocompetent patient with non-cystic fibrosis. The organism was identified by 16S rRNA gene sequencing. The patient was successfully treated with antibiotics.
Anti-Bacterial Agents ; Fibrosis* ; Genes, rRNA ; Humans ; Nontuberculous Mycobacteria ; Respiratory Tract Infections ; Sequence Analysis, RNA

BACKGROUND: Interferon-gamma has various biologic effects, including antiviral effect, antitumor proliferative effect, activation of macrophage and B lymphocyte, and increased expression of major histocompatibility complex. Especially, antitumor proliferative effect of interferon-gamma has already been proved to be important in vivo as well as in vitro. And, clinical studies of interferon-gamma have been tried in lung cancer patients. However, the mechanism of antitumor effect of interferon-gamma has not yet been established despite of many hypotheses. "Necrosis" is a type of cell death which is well known to occur in the circumstances of severe stresses. In contrast, "apoptosis" is another type of cell death which occurs in such biological circumstances as embryonic development, regression of organs, and self-tolerance of lymphocytes. And, apoptosis is an active process of cell death in which cells are dying with fragmentations of their cytoplasms and nuclei. And, in the process of apoptosis the DNAs of cells are cleaved between nucleosomes by unidentified endonuclease and therefore DNAs of apoptotic cells result in a typical electorphoresis pattern known as DNA ladder pattern. Recently it has been suggested that cytotoxic effect of interferon-gamma occurs via apoptosis. To elucidate the mechanism of antitumor cytotoxic effect of interferon-gamma, we microscopically observed a lung cancer cell line, A549 which was treated with interferon-gamma. We observed A545 treated with interferon-gamma was dying fragmented. And so, we performed this study to find out that the mechanism of antitumor cytotoxic effect of interferon-gamma be apoptosis. METHOD: We treated A549, human lung cancer cell line with various concentration of interferon-gamma and quantified its cytotoxic effect of various periods, 24 hours, 72 hours and,, 120 hours by MTT(dimethylthiazoly1 diphenyltetrazolium bromide) bioassay. Also, after we treated A549 with 100 units/ml of interferon-gamma for 120 hours, we observed the pattern of cell death with inverted microscope and we extracted DNAs from the dead A549 cells and observed the pattern of 1.5% agarose gel electrophoresis with ethidium bromide staining. RESULT: 1) Cytotoxic effect of interferon-gamma on A549: For the first 24 hours, threre was little cytotoxic effect and for between 24 hours and 72 hours, there was the beginning of cytotoxic effect and for 120 hours there was increased cytotoxic effect. 2) Pattern of A549 cell death by interferon-gamma: We observed with inverted microscope that A549 cells were dying fragmented. 3) DNA ladder pattern of gel electrophoresis: We observed DNA ladder pattern of gel electrophoresis of extracted DNAs from dead A549 cells. CONCLUSION: We concluded that the mechanism of interferon-gamma induced cytotoxicity on lung cancer cell line, A549 be via apoptosis.
Apoptosis ; Biological Assay ; Cell Death ; Cell Line* ; Cytoplasm ; DNA ; Electrophoresis ; Electrophoresis, Agar Gel ; Embryonic Development ; Ethidium ; Female ; Humans ; Interferon-gamma* ; Lung Neoplasms* ; Lung* ; Lymphocytes ; Macrophages ; Major Histocompatibility Complex ; Nucleosomes ; Pregnancy

Inflammatory bowel disease is associated with extraintestinal manifestations involving almost every organ system in the body. Crohn's disease (CD) appears to be more commonly associated with an inflammatory myopathy than ulcerative colitis. However, myopathy of the thigh in patients with CD is rare. We report an unusual site of necrotizing myositis in a patient with CD. A 23-year-old woman presented with swelling and pain at the left popliteal area that had lasted for 1 week. Twenty-two months before admission, she had presented with pyoderma gangrenosum on the left upper chest and was diagnosed with CD. A magnetic resonance imaging scan of her leg revealed diffuse swelling in the left semimembranous muscle and biceps femoris muscle that was compatible with myositis, and a cystic lesion in the distal portion of the semimembranous muscle. The findings from semimembranous muscle biopsy were also consistent with necrotizing myositis. In conclusion, myositis, although rare, can be an extraintestinal manifestation of CD.
Biopsy ; Colitis, Ulcerative ; Crohn Disease* ; Female ; Humans ; Inflammatory Bowel Diseases ; Leg ; Magnetic Resonance Imaging ; Muscles ; Muscular Diseases ; Myositis* ; Pyoderma Gangrenosum ; Thigh ; Thorax ; Young Adult

BACKGROUND: Goblet cell hyperplasia is a critical pathological feature in hypersecretory diseases of the airways. A bacterial infection of the lung is also known to induce inflammatory responses, which can lead to the overproduction of mucus. Recently, mucin synthesis in the airways has been reported to be regulated by neutrophilic inflammation-induced epidermal growth factor receptor (EGFR) expression and activation. In addition, it was reported that migration of the activated neutrophils is dependent on the matrix metalloproteinases (MMPs), especially MMP-9. In this study, bacterial lipopolysaccharide (LPS)-induced goblet cell hyperplasia and mucus hypersecretion by EGFR cascade, resulting from the MMPs-dependent neutrophilic inflammation were investigated in the rat airways. METHODS: Pathogen-free Sprague-Dawley rats were studied in vivo. Various concentrations of LPS were instilled into the trachea in 300microliter PBS (LPS group). Sterile PBS (300microliter) was instilled into the trachea of the control animals (control group). The airways were examined on different days after instilling LPS. For an examination of the relationship between the LPS-induced goblet cell hyperplasia and MMPs, the animals were pretreated 3 days prior to the LPS instillation and daily thereafter with the matrix metalloproteinase inhibitor (MMPI; 20 mg/Kg/day of CMT-3; Collagenex Pharmaceuticals, USA). The neutrophilic infiltration was quantified as a number in five high power fields (HPF). The alcian blue/periodic acid-Schiff (AB/PAS) stain were performed for the mucus glycoconjugates and the immunohistochemical stains were performed for MUC5AC, EGFR and MMP-9. Their expressions were quantified by an image analysis program and were expressed by the percentage of the total bronchial epithelial area. RESULTS: The instillation of LPS induced AB/PAS and MUC5AC staining in the airway epithelium in a time- and dose-dependent manner. Treatment with the MMPI prevented the LPS-induced goblet cell hyperplasia significantly. The instillation of LPS into the trachea induced also EGFR expression in the airway epithelium. The control airway epithelium contained few leukocytes, but the intratracheal instillation of LPS resulted in a neutrophilic recruitment. A pretreatment with MMPI prevented neutrophilic recruitment, EGFR expression, and goblet cell hyperplasia in the LPS-instilled airway epithelium. CONCLUSION: Matrix metalloproteinase is involved in LPS-induced mucus hypersecretion, resulting from a neutrophilic inflammation and EGFR cascade. These results suggest a potential therapeutic role of MMPI in the treatment of mucus hypersecretion that were associated with a bacterial infection of the airways.
Animals ; Bacterial Infections ; Coloring Agents ; Epidermal Growth Factor* ; Epithelium ; Glycoconjugates ; Goblet Cells ; Hyperplasia ; Inflammation ; Leukocytes ; Lung ; Matrix Metalloproteinases ; MMPI ; Mucins ; Mucus* ; Neutrophils* ; Rats ; Rats, Sprague-Dawley ; Receptor, Epidermal Growth Factor* ; Trachea