• Title/Summary/Keyword: 중합효소연쇄반응법

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Capsaicin induced apoptosis and the enhanced anticancer effect of anticancer drugs in cancer cells (종양세포에서의 capsaicin에 의한 apoptosis 유도와 항암제의 항암효과의 증가)

  • Kim, Sun Young;Lee, You Jin;Park, Eun Hye;Yi, Ho Keun;Jo, Dae Sun;Kim, Jung Soo;Hwang, Pyoung Han
    • Clinical and Experimental Pediatrics
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    • v.51 no.3
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    • pp.307-314
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    • 2008
  • Purpose : Capsaicin, the major pungent ingredient in red pepper, has long been used in spices and food additives. It has been recently shown to induce apoptosis in several cell lines through a not well known mechanism. The aim of this study was to investigate the apoptosis-inducing effect of capsaicin on gastric cancer cells, and to provide valuable information concerning the application of capsaicin for therapeutic purposes. Methods : Cultured SNU-668 cells were treated with capsaicin. We analyzed cell survival by trypan blue and crystal violet analysis, cell cytotoxicity by MTT assay, apoptosis by nuclear condensation and DNA fragmentation, bcl-2 and bax mRNA expression by RT-PCR, and the expression of apoptosis related proteins by Western immunoblot analysis. In order to assess whether the growth inhibitory effect of anticancer drugs is enhanced by capsaicin, we investigated the effects of cell cytotoxicity and the expression of apoptosis related proteins of etoposide and adriamycin treated with capsaicin in cells. Results : Capsaicin inhibited growth of SNU-668 cells in a dose-dependent manner. This inhibitory effect of capsaicin on cell growth was mainly due to the induction of apoptosis as evidenced by DNA fragmentation, nuclear condensation and the expression of apoptosis related proteins. Furthermore, capsaicin prominently reduced the ratio of anti-apoptotic Bcl-2 to pro-apoptotic Bax and consequently increased caspase-3 activity. The cells treated with capsaicin were more sensitive to death induced by etoposide and adriamycin than the cells without capsaicin. Conclusion : These results demonstrate that capsaicin efficiently induced apoptosis in SNU-668 cells through a caspase-3-dependent mechanism and sensitizes cancer cells to anticancer drugs toward apoptotic cell death, which may contribute to its anticancer effect and chemosensitizer function against gastric cancer.

Clinical Utility of Polymerase Chain Reaction for the Differentiation of Nontuberculous Mycobacteria in Patients with Acid-fast Bacilli Smear-positive Specimens (객담 항산균 도말 양성 환자에서 비결핵항산균과의 감별을 위한 결핵균 중합효소연쇄반응 검사의 유용성)

  • Lee, Jae Seung;Ji, Hyun Shuk;Hong, Sang Bum;Oh, Yeon-Mok;Lim, Chae-Man;Lee, Sang Do;Koh, Younsuck;Kim, Woo Sung;Kim, Dong Soon;Kim, Won Dong;Shim, Tae Sun
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.5
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    • pp.452-458
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    • 2005
  • Background : In Korea, polymerase chain reaction (PCR) test for M. tuberculosis has been used for the diagnosis of acid-fast bacilli (AFB) smear-negative tuberculosis in order to increase diagnostic sensitivity. However, there have been no data dealing with the clinical utility of PCR in AFB smear-positive patients to differentiate between M. tuberculosis and nontuberculous mycobacteria. Method : We retrospectively analyzed the PCR test results which have been performed in patients who had AFB smear-positive sputum but had ambiguous clinical manifestations of active tuberculosis. PCR test was done using $AMPLICOR^{\hat{a}}$ M. tuberculosis kit. The sensitivity, specificity, and positive and negative predictive values of the PCR test were calculated based on culture and final clinical diagnosis result. Results : Fifty-six consecutive patients (62 PCR tests) were included in the study. Active tuberculosis was diagnosed in 23 patients (41.0%), while 9 patients had NTM infection (16.0%). The sensitivity, specificity, positive- and negative-predictive value of PCR test were 88.8%, 86.8%, 76.1% and 94.3%, respectively, according to the culture result. In comparison, they were 91.3%, 100%, 100%, 94.3%, respectively, according to the final clinical diagnosis. All 15 patients with NTM isolates, including 6 patients who had other lung diseases but expectorated NTM isolate, were negative for PCR test. Conclusion : Even though tuberculosis is still prevalent in Korea, PCR test is useful to differentiate between M. tuberculosis and NTM in patients with AFB-smear positive sputum but with ambiguous clinical manifestations of active tuberculosis.

Clinical Characteristics of Mycoplasma pneumoniae Pneumonia in Children during Recent 10 Years (최근 10년간 Mycoplasma pneumoniae 폐염의 임상양상에 대한 고찰)

  • Lee, Gang Woo;Ryu, Ho Jun;Kim, Il Kyung;Seong, Ho;Choi, Chang Hee
    • Pediatric Infection and Vaccine
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    • v.6 no.1
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    • pp.86-92
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    • 1999
  • Purpose : The purpose of this study is to know the epidemiology and clinical characteristics in patients with M. pneumoniae pneumonia during study period retrospectively. Methods : We had reviewed 500 patients with Mycoplasma pneumoniae pneumonia who were hospitalized at the pediatric ward of Seoul Adventist Hospital from July 1986 through June 1996. we had analysed clinical and laboratory problems of pulmonary and extrapulmonary manifestations including demographics and epidemiological characteristics. Results : Peak occurrence of the disease was 3 years interval and seasonal occurrence of the disease had no difference. 43.4% of patients was below 4 years old and male to female ratio was nearly 1:1. The most common site of infiltration on chest PA was right lower lobe and the most common type of pneumonia was interstitial pneumonia(43%). Pulmonary complications were atelectasis(11cases), emphysema(5cases), pleural effusion(27cases) and sinusitis(13cases) and extrapulmonary manifestations were hepatitis(109cases), skin rash(29cases), proteinuria(20cases) and hematuria(16cases). cold agglutinin test was carried out in 500cases and mycoplasma antibody test was carried out in 448cases. sensitivity of mycoplasma antibody test was 53.3%. Conclusion : Mycoplasma pneumoniae pneumonia showed epidemics at intervals of 3 year in its major occurrence. it exhibits a tendency that occurs at younger age group not in school age group. diagnosis of the disease need more accurate method due to low sensitivity of mycoplasma antibody test. also we need to carried out other laboratory test(for example, LFT, U/A).

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Expression of nitric oxide synthase isoforms and N-methyl-D-aspartate receptor subunits according to transforming growth factor-β1 administration after hypoxic-ischemic brain injury in neonatal rats (신생 백서의 저산소 허혈 뇌손상에서 Transforming Growth Factor-β1 투여에 따른 Nitric Oxide Synthase 이성체와 N-methyl-D-aspartate 수용체 아단위의 발현)

  • Go, Hye Young;Seo, Eok Su;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.52 no.5
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    • pp.594-602
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    • 2009
  • Purpose : Transforming growth factor (TGF)-${\beta}1$ reportedly increases neuronal survival by inhibiting the induction of inducible nitric oxide synthase (NOS) in astrocytes and protecting neurons after excitotoxic injury. However, the neuroprotective mechanism of $TGF-{\beta}1$ on hypoxic-ischemic (HI) brain injury in neonatal rats is not clear. The aim of this study was to determine whether $TGF-{\beta}1$ has neuroprotective effects via a NO-mediated mechanism and N-methyl-D-aspartate (NMDA) receptor modulation on perinatal HI brain injury. Methods : Cortical cells were cultured using 19-day-pregnant Sprague-Dawley (SD) rats treated with $TGF-{\beta}1$ (1, 5, or 10 ng/mL) and incubated in a 1% O2 incubator for hypoxia. Seven-day-old SD rat pups were subjected to left carotid occlusion followed by 2 h of hypoxic exposure (7.5% $O_2$). $TGF-{\beta}1$ (0.5 ng/kg) was administered intracerebrally to the rats 30 min before HI brain injury. The expressions of NOS and NMDA receptors were measured. Results : In the in vitro model, the expressions of endothelial NOS (eNOS) and neuronal NOS (nNOS) increased in the hypoxic group and decreased in the 1 ng/mL $TGF-{\beta}1-treated$ group. In the in vivo model, the expression of inducible NOS (iNOS) decreased in the hypoxia group and increased in the $TGF-{\beta}1$-treated group. The expressions of eNOS and nNOS were reversed compared with the expression of iNOS. The expressions of all NMDA receptor subunits decreased in hypoxia group and increased in the $TGF-{\beta}1$-treated group except NR2C. Conclusion : The administration of $TGF-{\beta}1$ could significantly protect against perinatal HI brain injury via some parts of the NO-mediated or excitotoxic mechanism.

Characterization Analysis for Cardiogenic Potential of Three Human Adult Stem Cells (세 종류 줄기세포의 특성 분석과 지방유래 줄기세포의 심근세포로의 분화)

  • Park, Se-Ah;Kang, Hyeon-Mi;Kim, Eun-Su;Kim, Jin-Young;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.11 no.3
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    • pp.167-177
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    • 2007
  • In the present study, we isolated three human adult stem cells including adipose tissue-derived stem cells(HAD), umbilical cord-derived stem cells(HUC), and amnion-derived stem cells(HAM) and analysed their characteristics. And we examined whether HAD could be used as therapeutical cells for the heart diseases. Both HAM and HUC appeared very similar morphology but HAD was different. Doubling time of HUC was most fast, but total doubling numbers of HUC was same with HAM. Total doubling numbers of HAD was much more than others. Expression patterns of genes and proteins of three human adult stem cells were very similar. Also they were differentiated into adipocytes, osteocytes, and chondrocytes. In addition, they expressed many cardiomyocyte-related genes. But expression pattern of genes is a little different. When HAD were cultivated in the presence or absence of various combinations of BMP and FGF after 5-azacytidine expose for 24 h, expression of Cmlc-1, and ${\alpha}1c$ genes was significantly increased. However, expression of troponin T, troponin I and Kv4.3 genes was not changed. Based on these observations, it is suggested that HAD, HUC, and HAM might be used as potentially therapeutical cells for clinical application.

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DETECTION OF METHICILLIN OR VANCOMYCIN-RESISTANT STAPHYLOCOCCUS AUREUS FROM DENTAL HOSPITAL (치과병원 진료실 내에서 메티실린 또는 반코마이신 저항성 Staphylococcus aureus의 검출)

  • Min, Jung-Hee;Park, Soon-Nang;Hwang, Ho-Keel;Min, Jung-Beum;Kim, Hwa-Sook;Kook, Joong-ki
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.102-110
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    • 2007
  • The purpose of this study was to obtain the basic information for the improvement of dental environment by investigating the presence of methicillin- or vancomycin-resistant Staphylococcus aureus (MRSA or VRSA) isolated from dental health care workers (DHCWs) and environment of the Chosun University Dental Hospital (CUDH) and a private dental clinic (control group). Staphylococcus aureus (S. aureus) was isolated from anterior nares of 42 DHCWS and 38 sites, unit chairss, x-ray devices, computers, etc., at 10 departments of the CUDH and 20 DHCWs and 11 sites at the private dental clinic. S. aureus was isolated on mannitol salt agar plate and confirmed by PCR with S. aureus species-specific primer. Antimicrobial susceptibility test of clinical isolates of S. aureus against several antibiotics including methicillin (oxacillin) was performed by investigating minimum inhibitory concentration (MIC) using broth microdilution assay. In addition, PCR was performed to detect the methicillin- or vancomycin-resistant gene. The data showed that one strain of S. aureus was isolated from DHCWs of the CUDH and three strains of S. aureus was isolated from 3 samples of the private dental clinic, respectively. All of the isolates from the CUDH and the private dental clinic had resistance to penicillin G, amoxicillin and vancomycin and susceptibility to oxacillin and ciprofloxacin. The S. aureus strains were already obtained the resistance to penicillin G and amoxicillin. These results suggest that two dental clinics were under relatively safe environment.

How Reliable is Sputum PCR Test in the Diagnosis of Pulmonary Tuberculosis When Sputum Smear is Negative? (객담 결핵균 도말검사가 음성일때 중합효소연쇄반응검사와 진단적 신뢰도에 관한 연구)

  • Baek, Seung-Hoon;Lee, Jae-Myung;Kang, Min-Jong;Son, Jee-Woong;Lee, Seung-Joon;Kim, Dong-Gyu;Lee, Myung-Goo;Hyun, In-Gyu;Jung, Ki-Suck;Lee, Kyung-Wha;Joe, Hyun-Chan
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.2
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    • pp.222-228
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    • 2001
  • Backgrounds : Recent technological developments have introduced a new method to identifying M. tuberculosis complex DNA in clinical samples directly. The direct amplification test (DAT) is approved for identifying M. tuberculosis complex in respiratory specimens that are smear-positive for acid-fast bacilli (AFB). When there is a discrepancy between the AFB smear and DAT, no information on their clinical utility is currently available. In this study, the diagnostic reliability of DAT was investigated in suspected pulmonary tuberculosis patients whose sputum AFB smear was negative. Methods : From June 1, 1998 through May 30, 1999, 909 patients with presumed active pulmonary tuberculosis were enrolled. A sputum AFB stain, culture, DAT and/or biopsy were performed. Using the criteria of clinical tuberculosis or confirmed tuberculosis, the positive predictive value of DAT in diagnosing pulmonary tuberculosis was investigated. Results : The positive predictive value of DAT was 82.1% by the clinically active tuberculosis criteria. However, it decreased to 61.5% when diagnosis was restricted to only to culture positive or biopsy proven cases. The false positive rate of DAT was 18.0%. Conclusion : The DAT is a valuable diagnostic method in suspected patients whose sputum AFB is was negative.

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Analysis of Bone Marrow Micrometastasis Using RT-PCR in Patients with Small Cell Lung Carcinoma (소세포 폐암에서 역전사 중합효소 연쇄반응(RT-PCR)을 이용한 골수 미세전이(naicronaetastasis)의 분석)

  • Kim, Tae-You;Park, Jong-Kook;Ryoo, Baek-Ryeol;Im, Yung-Hyuck;Kang, Yoon-Koo
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.6
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    • pp.797-806
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    • 1999
  • Background: About 20% of small cell lung cancer(SCLC) patients have bone marrow(EM) metastasis at the time of diagnosis and the remaining patients are also considered with micrometastasis. In an attempt to detect EM micrometastasis, we used cytokeratin(CK)-20 as a molecular marker, which is specific for epithelial cells. Method: A sensitive RT-PCR assay was used to compare CK-20 expression both in SCLC cell line H209 and normal leukocyte and to evaluate EM aspirates of 28 SCLC patients. Result: H209 cell line showed CK-20 expression but normal leukocyte did not, suggesting CK-20 expression is lung tissue-specific. Of 28 patients(11 limited disease, 17 extensive disease), only 2(1/11, 1/17) samples tested revealed positive signal for CK-20. Two patients with CK-20 expression had EM metastasis or multiple bone involvement during follow-up. Conclusion: Although circulating tumor cells were detected in EM of small portion of patients with bone metastasis, CK-20 doesn't seem to be a reliable marker for the detection of micrometastasis in SCLC. This study emphasizes that identification of more specific marker for micrometastsis is mandatory prior to clinical application.

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Inhibition of Viability and Genetic Change in Hypoxia-treated Lung Pericytes (허파혈관주위세포에서 저산소증에 의한 생존능의 억제와 유전자 발현의 변화)

  • Shin, Jong Wook;Kim, Kae-Young;Lee, Young Woo;Jung, Jae Woo;Lee, Byoung Jun;Kim, Jae-Yeol;Jo, Inho;Park, In Won;Choi, Byoung Whui
    • Tuberculosis and Respiratory Diseases
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    • v.57 no.1
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    • pp.37-46
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    • 2004
  • Background : Lung pericytes are important constituent cells of blood-air barrier in pulmonary microvasculature. These cells take part in the control of vascular contractility and permeability. In this study, it was hypothesized that change of lung pericytes might be attributable to pathologic change in microvasculature in acute lung injury. The purpose of this study was how hypoxia change proliferation and genetic expression in lung pericytes. Methods : From the lungs of several Sprague-Dawley rats, performed the primary culture of lung pericytes and subculture. Characteristics of lung pericytes were confirmed with stellate shape in light microscopy and immunocytochemistry. 2% concentration of oxygen and $200{\mu}M$ $CoCl_2$ were treated to cells. Tryphan blue method and reverse transcription-polymerase chain reaction were done. Results : 1. We established methodology for primary culture of lung pericytes. 2. Hypoxia inhibited cellular proliferation in pericytes. 3. Hypoxia could markedly induce vascular endothelial growth factor(VEGF) and smad-2. 4. Hypoxia-inducible factor-$1{\alpha}$(HIF-$1{\alpha}$) was also induced by 2% oxygen. Conclusion : Viability of lung pericytes are inhibited by hypoxia. Hypoxia can stimulate expression of hypoxia-responsive genes. Pericytic change may be contributed to dysfunction of alveolar-capillary barrier in various pulmonary disorders.