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Comparison of PCR-Line Probe and PCR-SSCP Methods for the Detection of Rifampicin Resistant Mycobacterium Tuberculosis (Rifampicin 내성 결핵균의 검출에 있어서 PCR-line Probe법과 PCR-SSCP법의 비교)

  • Kim, Ho-Joong;Suh, Gee-Young;Chung, Man-Pyo;Kim, Jong-Won;Shim, Tae-Sun;Choi, Dong-Chull;Kwon, O-Jung;Rhee, Chong-H;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.714-722
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    • 1998
  • Background: Rifampicin (RFP) is a key component of the antituberculous short-course chemotherapy and the RFP resistance is a marker of multi-drug resistant (MDR) tuberculosis. RPoB gene encodes the $\beta$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. And the mutations of rpoB gene have been found in about 96% of rifampicin resistant clinical isolates of M. tuberculosis. So in order to find a rapid and clinically useful diagnostic method in identifying the RFP resistance, we compared the PCR -line probe method with PCR-SSCP for the detection of the rpoB gene mutation in cultured M. tuberculosis. Methods: 45 clinical isolates were collected from patients who visited Sung Kyun Kwan University Hospital. The RFP susceptibility test was referred to the referral laboratory of the Korean Tuberculosis Institute. 33 were rifampicin resistant and 12 were rifampicin susceptible. The susceptibility results were compared with the results of the PCR-BSCP and PCR-line probe method. Results: We could find rpoB mutations in 27/33(81.8%) RFP-resistant strains by PCR-line probe method, and in 23/33 (69.7%) by PCR-SSCP and there was no significant difference between two methods. There was no mutation in rifampicinn susceptible strains by both methods. Conclusion: PCR-line probe method would be a rapid, sensitive and specific method for the detection of rifampicin resistant Mycobacterium tuberculosis.

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The effect of 8 French catheter and chest tube on the treatment of spontaneous pneumothorax (자연기흉에 있어서 8 French 도관과 흉관의 삽입 치료 효과)

  • Kang, Yoon-Jeong;Koh, Hyoung-Gee;Shin, Jong-Wook;Lim, Seong-Yong;Choi, Jae-Sun;Yu, Ji-Hoon;Park, In-Won;Choi, Byoung-Whui;Hue, Sung-Ho;Seo, Seung-Cheon
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.3
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    • pp.410-419
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    • 1996
  • Background : Spontaneous pneumothorax have been managed with a variety of methods. The technique most frequently used is chest tube drainage. Small caliber catheters were first used in the management of pneumothorax complicating the percutaneous needle aspiration lung biopsy, and the try to treat spontaneous pneumothorax also has been reported. However, the value of small caliber catheters in spontaneous pneumothorax has not been fully evaluated. So, we tried to elucidate the efficacy of 8 French catheter in the management of spontaneous pneumothorax. Method : From January, 1990, to April, 1994, 44 patients with spontaneous pneumothorax treated at Chung-Ang university hospital were reviewed. The patients were sub-divide into 8 French catheter insertion group (n=21) and chest tube insertion group (n=23). We compared the presence of underlying lung disease, the extent of the collapse, the duration of indwelling catheter and complication between two groups. Results : 1) The duration of indwelling showed no significant difference between 8 French catheter group and chest tube. But, complication after insertion as subcutaneous emphysema was developed in only chest tube group. (p<0.05) 2) In the primary spontaneous pneumothorax, all case of the pneumothorax of which size was less than 50% showed complete healing with 8 French catheter insertion. Whereas the success rate in patients with large pneumothorax (more than 50%) was tended to be dependent on the age. 3) In the patients with secondary spontaneous pneumothorax who were managed with 8 French catheter, the success rate was trended to be high if the underlying disease of pneumothorax was not COPD and if the patient was young. Conclusion : These results show that 8 French catheter insertion probably was effective in the pneumothorax less than 50%, the primary spontaneous pneumothorax, young age or secondary pneumothorax not associated with COPD.

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Association of Genetic Polymorphism of Glutathione S-transferase M1, T1 and N-acetyltransferase 1 with Lung Cancer (폐암발생과 Glutathione S-transferase M1, T1 및 N-acetyltransferase 1의 유전적 다형성과의 연관성에 관한 연구)

  • Lee, Seung-Joon;Park, Gye-Young;Oh, Yeon-Mok;Kang, Dae-Hee;Cho, Soo-Hun;Kim, Soo-Ung;Yoo, Chul-Gyu;Lee, Chun-Taeck;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.4
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    • pp.471-477
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    • 1999
  • Background : Smoking and high-risk occupation have been known to be the risk factors of lung cancer. The carcinogen-metabolizing enzymes in human body such as glutathione S-transferase M1, T1 and N-acetyltransferase 1 have also been regarded as risk factors in many cancers, because the activities of those enzymes play a role in metabolizing the carcinogen. A case-control study was conducted to evaluate the genetic polymorphism of GSTM1, T1 and NAT1 in lung carcinogenesis in Korean men. Methods : The histologically proven lung cancer cases were recruited from Seoul National University Hospital. The patients of more than 40-year-old with the nonmalignant urinary tract diseases were recruited as controls from the same hospitals. The informations of demographical characteristics and smoking were obtained by interview or chart review and the genetic polymorphisms of GSTM1, T1 and NAT1 were determined by PCR-based assay. The statistical analyses were performed by linear logistic regression. Results : The number of case-control was 118 and 150, respectively. The smoking history was significantly higher in the lung cancer patients than the controls. The prevalence of GSTM1 null-type was statistically higher(OR=2.25 ; 95% CI=1.12-4.51) in squamous cell carcinoma than other genotypes, but other histologic types were not The prevalence of GSTT1 null-type were not statistically higher than other genotypes in all histologic types. The fast acetylator of NAT1 was more prevalent than normal(OR=2.13 ; 95% CI=1.04-4.40) in all lung cancer patients. Conclusion : The null-type of GSTM1 and fast acetylator of NAT1 are associated with development of lung cancer in Korean men.

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Polymorphisms of GSTM1 and CYP1A1, and Susceptibility to Primary Lung Cancer in Korean Males (한국인 남성에서 GSTM1과 CYP1A1 유전자 다형성과 원발성폐암의 유전적 감수성)

  • Bae, Nack-Cheon;Lee, Su-Yeon;Chae, Po-Hee;Kang, Kyung-Hee;Kim, Kyung-Rock;Cha, Seung-Ick;Chae, Sang-Chul;Kim, Chang-Ho;Jung, Tae-Hoon;Park, Jae-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.5
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    • pp.568-578
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    • 2001
  • Background : Lung cancer is frequently cited as an example of a disease caused solely by exposure to environmental carcinogens. However, there is a growing realization that the genetic constitution is also important in determining individual's susceptibility to lung cancer. This genetic susceptibility may result from functional polymorphims of the genes involved in carcinogen metabolism. In this study, the association between GSTM1 and CYP1A1 polymorphisms and the lung cancer risk in Korean males was investigated. Materials and Method : The study population consisted of 153 male lung cancer patients and 143 healthy male controls. The GSTM1 and CYP1A1 genotypes were determined by multiplex PCR and PCR-RFLP analysis. Result : There were no significant differences in the frequency of the GSTM1 null genotype between the cases and the controls. When the cases were categorized by their histologic type, the frequency of the GSTM1 null genotype in the small cell carcinoma group was higher than those of the controls(67.2% vs 55.9%), but the difference was not statistically significant(OR=1.772 ; 95% CI=0.723-4.340). The distribution of the CYP1A1 MspI genotypes among the cases were similar to those among the controls. When the cases were grouped by their histologic type, the ml/m1, ml/m2, m2/m2 genotypes frequencies among the small cell carcinomas(23.0%, 38.5%, and 38.5%, respectively) were significantly different from those of the controls(36.4%, 46.2%, and 7.4%, respectively, p<0.05). When the m1/m1 genotype was used as a reference, the ml/m2 and m2/m2 genotypes were associated with an increased risk for small cell lung cancer(ml/m2 genotype : OR=1.337, 95% CI=0.453-3.947 ; m2/m2 genotype : OR=3.374, 95% CI=1.092-10.421). Conclusion : These results suggest that the GSTM1 and CYP1A1 genotypes may be a genetic determinant of the risk for lung cancer, particlulary small cell carcinoma. Further investigation is needed to confirm these results.

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Relation Among Parameters Determining the Severity of Bronchial Asthma (기관지천식 환자의 증상의 중증도를 나타내는 지표들간의 연관성)

  • Lee, Sook-Young;Kim, Seung-June;Kim, Seuk-Chan;Kwon, Soon-Suk;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Song, Jeong-Sup;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.5
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    • pp.585-593
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    • 2000
  • Background : International consensus guidelines have recently been developed to improve the assessment and management of asthma. One of the major recommendation of these guidelines is that asthma severity should be assessed through the recognition of key symptoms, such as nocturnal waking, medication requirements, and objective measurements of lung function. Differential classification of asthma severity would lead to major differences in both long term pharmacological management and the treatment of severe exacerbation. Methods : This study examined the relationship between the symptom score and measurements of $FEV_1$ and PEF when expressed as a percentage of predicted values in asthmatics (n=107). Results : The correlation of $FEV_1$ % with PEFR% was highly significant (r=0.83, p<0.01). However, there was agreement in terms of the classification of asthma severity in 76.6% of the paired measurements of $FEV_1$ % and PEFR%. Agreement in the classification of asthma severity was also found in 57.1% of the paired analysis of $FEV_1$ % and symptom score. 39% of the patients classified as having moderate asthma on the basis of $FEV_1$ % recording would be considered to have severe asthma if symptom score alone were used. Low baseline $FEV_1$ and high bronchial responsiveness were associated with a low degree of perception of airway obstruction. Conclusion : The relationships between the symptom score, PEFR and $FEV_1$ were generally poor. When assessing asthma severity, age, duration, $PC_{20}$, and baseline $FEV_1$ should be considered.

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Adherence-induced gene expression in human alveolar macrophages (표면부착에 의한 사람 폐포대식세포의 유전자 발현에 관한 연구)

  • Chung, Man Pyo;Yoo, Chul Gyu;Han, Sung Koo;Shim, Young-Soo;Rhee, Chong H.;Han, Yang Chol;Kim, Young Whan
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.936-944
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    • 1996
  • Background: Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. Adhesion molecules and gene transcription of the inflammatory mediators are known to be associated in this process. To evaluate whether adhesion molecule and transcriptional activation of the inflammatory substances are also involved in the activation of human alveolar macrophage by the adherence procedure, we designed this experiment. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be nonnal by chest cr and alveolar macrophage was harvested. To measure the expression of Interleukin-8(IL-8) mRNA, manganese superoxide dismutase(SOD) mRNA and CD11/CD18 mRNA in human alveolar macrophage of both adherence state and suspension state, Northern blot analysis was done at 0, 2, 4, 8 and 24hrs after the adherence to plastic surface and during suspension state. Then, phorbol myristate acetate(pMA) and N-formyl-methionyl-leucyl-phenylalanine(fMLP) were added respectively in the same experimental condition. Result : 1) Human alveolar macrophages in the adherent state induced IL-8 mRNA and SOD mRNA expression which was maximal at 8 hours after the adherence to plastic surface. But we could not observe the upregulation of CD18 mRNA by surface adherence. 2) PMA induced these mRNA expression both in the adherent cell and the nonadherem cells, but the induction of mRNA expression by fMLP occurred only in the adherent cells. Conclusion: These results suggest that adherence of huamn alveolar macropahge is an important cell-activating event that may play a critical role in the modulation of lung inflammatory respones.

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Rapid detection of Rifampicin- resistant M, tuberculosis by PCR-SSCP of rpoB gene (결핵균의 rpoB유전자 PCR-SSCP법에 의한 Rifampicin 내성의 신속 진단)

  • Shim, Tae Sun;Yoo, Chul-Gyu;Han, Sung Koo;Shim, Young-Soo;Kim, Young Whan
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.842-851
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    • 1996
  • Background : Rifampicin(RFP) is a key component of the antituberculous shon-course chemotherapy and the RFP-resistance is a marker of multi-drug resistant(MDR) M. tuberculosis. rpoB gene encodes the ${\beta}$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. Recent reports show that rpoB gene mutations are the cause of RFP resistance of M. tuberculosis and the main mechanism of rpoB gene mutation is point mutation. And PCR-SSCP is a rapid and easy method for detecting point mutations. So we performed PCR-SSCP of rpoB gene of M. tuberculosis and compared the result with traditional RFP sensitivity test. Method : The 27 RFP sensitive M. tuberculosis culture isolates and 25 RFP resistant isolates were evaluated. The RFP sensitivity test was done at the Korean Tuberculosis istitute. The DNA was extracted by bead beater method and was amplified with primers TR-8 and TR-9 in a 20ul PCR reaction containing 0.1ul(luCi) [${\alpha}-^{32}P$] - dCTP. After amplification, SSCP was done using non-denaturaring polyacrylamide gel electrophoresis. Then direct sequencing was done in cases of different eletrophoretic mobility compared with that of H37Rv. In 19 cases, we compared PCR-SSCP results with patient's clinical course and the results of traditional RFP sensitivity test. Results : 1) All 27 RFP sensitive M. tuberculosis isolates showed the same electrophoretic mobility compared with that of H37Rv. And all 25 RFP resistant M. tuberculosis isolates showed different electrophoretic mobility. 2) The mechanism of rpoB gene mutation of M. tuberculosis is mainly point mutation. 3) The PCR-SSCP results correlate well with traditional RFP sensitivity and patient's clinical response to antituberculous treatment. Conclusion: The PCR-SSCP of rpoB gene is a very sensitive and rapid mehod in detecting RFP- resistant M. tuberculosis.

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Aberrant Promoter Methylation of Death-Associated Protein Kinase in Serum DNA from Lung Cancer Patients (원발성 폐암 환자의 혈청에서 DAP kinase 유전자의 Methylation 양상)

  • Lee, Jun Hee;Lee, Jung Wook;Jung, Kyung Sik;Kim, Ki Uk;Lee, Tae Kun;Lee, Kyung Woo;Na, Min-Ah;Jeon, Doo Soo;Choi, Young Min;Kim, Yun Seong;Lee, Min Ki;Park, Soon Kew
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.4
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    • pp.378-387
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    • 2003
  • Background : Promoter methylation of tumor suppressor genes is one of the key epigenetic changes in many human cancers. The aim of this study was to evaluate the promoter methylation status of the Death-associated protein(DAP) kinase gene, which played an important role in lung cancer, in the serum DNA of primary lung cancer patients. Methods : This study investigated the aberrant methylation of DAP kinase in the serum of 65 primary lung cancer patients by methylation-specific PCR (MSP). Results : Methylation in the serum was detected in 29 of 65(44.6%) for DAP kinase. There was no statistical association between methylation of DAP kinase and age, smoking history, histologic type, or stage. Methylation of DAP kinase was found more frequently in men (p=0.044). Conclusions : This study suggests that the aberrant methylation of the DAP kinase promoter is readily detectable in the serum DNA of lung cancer patients using MSP analysis.

Clinical Investigation of Recurrent Pneumonia in Adults - Analysis of Patients From Hallym University Medical Center - (성인 재발성 폐렴에 대한 임상적 고찰 - 한림대학교 의료원 내원 환자를 대상으로 -)

  • Eom, Kwang-Seok;Jeon, Gang;Shin, Taerim;Jang, Seung Hun;Bahn, Joon-Woo;Lee, Jae Young;Park, Yong Bum;Kim, Cheol Hong;Jeon, Man-Jo;Park, Sang Myeon;Kim, Dong Gyu;Lee, Myung Goo;Hyun, In-Gyu;Jung, Ki-Suck
    • Tuberculosis and Respiratory Diseases
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    • v.57 no.1
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    • pp.47-54
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    • 2004
  • Background : Recurrent pneumonia in adults is not uncommon. However, there is no domestic data about recurrent pneumonia in adults. Therefore, we investigated the associated diseases and clinical findings of recurrent pneumonia in adults. Methods : Among 5513 patients who were treated in five teaching hospitals of Hallym medical center?over a 5-year period, we retrospectively reviewed the medical records of the 58 who were compatible with diagnostic criteria of recurrent pneumonia. Results : The number of patients with recurrent pneumonia was 58 (1.05%, 58/5513) during the 5 years. Thirtyseven patients were male and 21 were female. Mean age was 66.4 (${\pm}14.9$) years. Median interval between each pneumonic episode was 18.5 months. Associated diseases were 25 cases of respiratory diseases, 13 of heart diseases, 13 of diabetes mellitus, 7 of lung malignancies, 11 of malignancies other than lung, 7 of neurologic disease, and 8 of miscellaneous diseases. Three cases had no underlying illness. Of the 8 cases with 2 or more times of recurrence, 4 were associated with respiratory diseases, 2 with aspiration pneumonia due to neurologic diseases, 1 with heart disease and 1 with no underlying illness. Recurrent pneumonic episodes affecting the same location were 30 of the total recurrent pneumonic episodes (30/67, 47.8%) and common associated diseases were respiratory diseases including lung malignancies. The etiology of recurrent pneumonia was Streptococcus pneumoniae, methicillinresistant Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, atypical organisms, etc. Conclusion : Recurrent pneumonia in adults had a low incidence rate compared with children, but most cases had associated illness. Respiratory diseases including lung cancer were the most common associated illness of recurrent pneumonia.

Food Component Characteristics of Red-tanner Crab (Chionoecetes japonicus) Paste as Food Processing Source (식품가공소재로서 게 페이스트의 식품성분 특성)

  • Kim, Hye-Suk;Park, Chan-Ho;Choi, Seung-Geal;Han, Byung-Wook;Kang, Kyung-Tae;Shim, Nam-Hyuk;Oh, Hyeon-Seok;Kim, Jin-Soo;Heu, Min-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.1077-1081
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    • 2005
  • The objective of this study was to investigate the components and nutritional quality of red-tanner crab (Chionoecetes japonicus) paste in order to explore possibility for food processing source such as surimi gel containing crab paste. Yield of crab paste was $30\%$ from whole body after crushing and dehydrating. Crude protein contents $(9.5\%)$ of crab paste was lower than that $(13.1\%)$ of crab muscle, but fat $(0.5\%)$ and ash contents $(8.0\%)$ of paste were higher than $0.2\%\;and\;1.3\%$ of crab muscle, respectively. Volatile basic nitrogen (VBN) content of the crab paste was lower than those of the edible parts. Total amino acid content (9,497mg/l00g) of paste was lower than that (12,980mg/100g) of muscle. Aspartic acid, glutamic acid, lysine and leucine were the predominant amino acids in the protein fraction. The calcium content (6,539mg/l00g) was higher than those of phosphorus (579mg/100g), and potassium (793mg/100g) while manganese and iron were present in trace amounts. Major fatty acids of total lipid were 16:0, 18:1n-9, 20:5n-3 and 22:6n-3, and no difference of composition between paste and muscle. Sensory evaluation showed that scores of color and flavor of $15\%$ substituted surimi gel increased significantly when compared to surimi gel without crab paste (p<0.05). From the above results, the addition of crab paste enhanced nutrition and functionality of surimi gel.