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      • KCI등재

        Carbapenemase를 생산하는 imipenem 내성 세균의 특성 및 항생제 감수성

        최한나(Han Na Choe),박철(Chul Park),김형락(Hyung Rak Kim),백근식(Keun Sik Baik),김세나(Se Na Kim),성치남(Chi Nam Seong) 한국생명과학회 2010 생명과학회지 Vol.20 No.8

        대한민국 순천의 병원 입원 환자의 검체로부터 imipenem 내성 세균을 분리하였다. 54개의 분리균을 16S rRNA 유전자와 gyrB 유전자 염기서열 비교를 기초로 하여 계통분류학적으로 동정하였다. 분리균들은 Pseudomonas aeruginosa (30균주; 55.6%), Acinetobacter baumannii (21; 38.9%), Enterobacter hormaechei (2)와 Pseudomonas putida (2)에 속했다. 22개의 균주가 metallo-β-lactamase (MBL)를 생산하였으며 종별 구성은 다음과 같다; Acinetobacter baumannii 12균주, Pseudomonas aeruginosa 7균주, P. putida 2균주 그리고 Enterobacter hormaechei 1균주. 분리균들의 항생제 감수성은 디스크 확산법과 Vitek 을 이용하여 조사하였다. IMP 와 VIM 형의 metallo-β-lactamase를 생산하는 균주들은 OXA 와 SHV 형 β-lactamase를 생산하는 균주들에 비해 ceftazidime, aztreonam, amikacin과 gentamicin에 대한 내성율이 높았다. Imipenem-resistant bacteria were isolated from clinical specimens taken from hospitalized patients in Suncheon, Korea. Fifty-four isolates were phylogenetically analyzed based on 16S rRNA gene and gyrB gene sequence comparisons. Isolates were affiliated with Pseudomonas aeruginosa (30 strains; 55.6%), Acinetobacter baumannii (21; 38.9%), Enterobacter hormaechei (2) and Pseudomonas putida (2). Twenty-two isolates produced metallo-β-lactamase (MBL); 12 Acinetobacter baumannii strains, 7 Pseudomonas aeruginosa strains, 2 P. putida strains and 1 Enterobacter hormaechei strain. Antibiotic susceptibility of the isolates was determined using the disc diffusion method and Vitek system. Strains producing metallo-β-lactamase (type IMP & VIM) were more resistant to antibiotics ceftazidime, aztreonam, amikacin and gentamicin than to strains producing OXA and SHV type of β-lactamase.

      • KCI등재

        류마티스관절염과 전신홍반루푸스 환자에서의 대유행 H1N1 인플루엔자 백신 접종 반응

        김지헌 ( Ji Hun Kim ),최한나 ( Han Na Choi ),김시혜 ( Si Hye Kim ),이화정 ( Hwa Jeong Lee ),박성훈 ( Sung Hoon Park ),김성규 ( Seong Kyu Kim ),최정윤 ( Jung Yoon Choe ),권현희 ( Hyun Hee Kwon ),정희진 ( Hee Jin Cheong ) 대한류마티스학회 2013 대한류마티스학회지 Vol.20 No.2

        Objective. We investigated the efficacy and safety of pandemic H1N1 vaccine in rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) patients, and evaluated its correlation with serum cytokine level. Methods. A total of 43 RA patients and 31 SLE patients were enrolled in the study and were compared with age, sex-matched 40 healthy controls (HC). The blood samples drawn from selected patients before vaccination and in post-vaccination at week 4 were assayed in one session to measure the titers of antibodies against haemagglutinin specific for influenza virus strains: A/California/7/2009 NYMC X-179A (H1N1). Serum IL17 and CXCL13 levels were measured in the same session by enzyme-linked immunosorbent assay. The association of serum cytokine level with anti-influenza antibody titer and mean fold increase (MFI) was investigated. Each specific side effect after vaccination was monitored in both the patients and control groups. Results. The geometric mean antibody titer (GMT) for preand post-vaccination at week 4 was not significantly different between RA and HC, SLE and HC. The seroconversion rate in HC and RA was not significantly different, whereas the seroprotection rate is significantly higher in HC (82.5%) than RA (55.8%) (p<0.05). MFI in HC, RA, SLE were 19.65, 6.00 and 6.06, which were significantly higher in HC. Serum IL17 level was 6.28±2.89 pg/mL and 7.56±3.34 pg/mL in pre-, post-vaccination SLE patients, 33.85±15.62 pg/mL and 38.04±18.60 pg/mL in RA patients and was significantly lower in SLE patients. Serum CXCL13 level was 518.73± 720.29 pg/mL and 431.53±601.23 pg/mL in pre-, post-vaccination SLE patients, which was significantly higher than HC (149.64±248.81 pg/mL and 147.36±213.92 pg/mL in each pre-, post-vaccination) and was not significantly different with the level of RA patients. In SLE patients, significant correlations were detected between cytokine level and post-vaccination antibody titer (r=0.22 p=0.026 between IL 17 and GMT; r=0.44, p<0.05 between CXCL13 and GMT). Conclusion. The increase in post-vaccination antibody titer is weaker in both RA and SLE patients group than the HC group. Post-vaccination antibody titer was positively correlated with B lymphocyte chemoattractant and CXCL13 in SLE patients, but not in RA patients.

      • KCI등재

        임상검체로부터 분리된 methicillin 내성 Staphylococcus aureus의 독소 및 항생제 내성

        Keun Sik Baik(백근식),Gwang Seo Ki(기광서),Han Na Choe(최한나),Seong Chan Park(박성찬),Eun Cho Koh(고은초),Hyung Rak Kim(김형락),Chi Nam Seong(성치남) 한국생명과학회 2011 생명과학회지 Vol.21 No.2

        2009년 7월부터 12월까지 순천 소재 한 병원에 내원한 환자의 검체로부터 methicillin 내성 Staphylococcus aureus(MRSA) 75균주와 methicillin 감수성 S. aureus (MSSA) 24균주를 분리하였다. 분리균의 항생제 감수성 조사는 디스크 확산법을 사용하여 측정하였다. 분리균의 독소 유전자 보유는 multiplex PCR을 이용하여 장독소(enterotoxin; SE), 독성 쇼크 증상 독소 1(toxic shock syndrome toxin-1; TSST-1), 피부박탈성 독소(exfoliative toxin; ET) 및 백혈구 용해 독소(Panton-Valentine leukocidin; PVL) 유전자를 검출하였다. 분리된 MRSA 60개 균주는 1개 혹은 2개의 독소 유전자를 가지고 있으며, 22.7%의 균주가 seb, sec, seg, sei와 tst 유전자를 동시에 보유하고 있었으며 18.7%는 sec, seg, sei와 tst 유전자를 동시에 보유하고 있었다. 백혈구 용해독소를 암호하는 pvl 유전자는 검출되지 않았다. MRSA는 sec, seg, sei와 tst 유전자 보유에 높은 상관성을 보였다. MRSA 균주들은 erythromycin(분리균의 89%), gentamicin (70.7%), ciprofloxacin (69.3%), clindamycin (61.3%)과 tetracycline (58.7%)에 내성이 높은 반면, MSSA 균주들은 erythromycin를 제외한 다른 항생제에는 민감하였다. 독소 유전자 seb, sec와 tst는 tetracycline 내성과 높은 상관관계가 있었다. Seventy five methicillin- resistant Staphylococcus aureus (MRSA) strains and 24 methicillin- susceptible S. aureus (MSSA) were isolated from clinical specimens obtained from a hospital in Suncheon, Jeonnam province, Korea, from July to December, 2009. Antibiotic resistance was determined using the disc diffusion method. Genes encoding enterotoxin (SE), toxic shock syndrome toxin-1 (TSST-1), exfoliative toxin (ET) and Panton-Valentine leukocidin (PVL) were detected by multiplex PCR-mediated amplification using specific primers. Sixty (80%) MRSA isolates possessed either one or more toxin genes and the most common pattern that coexisted in MRSA was seb, sec, seg, sei and tst (22.7%) followed by coexistence of sec, seg, sei and tst genes (18.7%). Gene pvl encoding leukocidin was not found. Significant correlation between the production of sec, seg, sei and tst genes was found. MRSAs were resistant to erythromycin (89% of the isolates), gentamicin (70.7%), ciprofloxacin (69.3%), clindamycin (61.3%) and tetracycline (58.7%), while MSSAs were susceptible to the antibiotics with the exception of erythromycin. Toxin genes seb, sec and tst were related to the tetracycline resistance of MRSA.

      • KCI등재

        Real-time PCR을 이용한 가축생균제용 유산균 정량분석

        Yeon Jae Choi(최연재),Sun Ho Kim(김선호),Min Jeong Gu(구민정),Han Na Choe(최한나),Dong Un Kim(김동운),Sang Bum Cho(조상범),Su Ki Kim(김수기),Che Ok Jeon(전체옥),Gui Seok Bae(배귀석),Sang Seok Lee(이상석) 한국생명과학회 2010 생명과학회지 Vol.20 No.12

        본 연구는 가축생균제용 유산균을 Real-time PCR정량분석법을 이용하여 분석하였다. SYBR Green1 방법과 Probe 방법을 이용하여 표준곡선을 제작한 결과, SYBR Green1 방법에서는 Slope 값이 -3.346이었고, Y절편은 33.18, R² 값은 0.993으로 나타났으며, Probe 방법에서는 Slope값이 -3.321이었고, Y절편은 39.10, R² 값은 0.995로 나타나, 이를 이용한 표준곡선 제작이 가능함을 알 수 있었다. SYBR Green1 방법을 이용한 생균제의 Lactobacilli 정성ㆍ정량 분석결과 Real-time PCR값은 4.46~6.56 log copies로 나타났고, 생균수 측정 결과 값은 5.63~7.59 log CFU/g로 나타났으며, Probe 방법을 이용한 생균제의 Lactobacilli 정성ㆍ정량 분석결과에서는 Real-time PCR 값은 5.51~7.00 log copies로 나타났으며, 생균수 측정 결과 값은 5.63~7.59 log CFU/g로 나타났다. 본 연구에서 실시한 RT PCR법은 3~4일이 소요되는 기존의 배지법과 비교하여 24시간 이내에 신속하게 검출이 가능하다고 여겨지며, 또한 RT PCR을 이용한 분석방법에서도 dye 사용과 primer 사용에 따라 결과값이 차이가 나타났음을 확인할 수 있었으며, Probe 방법을 이용하여 실험 한 결과가 민감한 결과를 나타내었음을 확인 할 수 있었다. This study was conducted using quantitative real-time PCR using Lactobacilli as probiotics. Quantitative real-time PCR (RT PCR) was conducted via a method involving SYBR Green 1 and a probe. Plasmid DNA was cloned using the 16S-23S rRNA intergenic species region. Gene clones were diluted from 10² to 10¹?. Standard curves were constructed via Ct values obtained from the results of Real-time PCR via the aforementioned SYBR Green 1 and probe method. Plasmid DNA was also cloned using the 16S-23S rRNA intergenic species region and the gene clones were diluted from 10² to 10¹? copy numbers via the probe method. Using RT PCR, a standard curve of plasmid DNA copy numbers was also determined. The slope value for the Y-axis intercept and R² value were measured as -3.346, 33.18, and 0.993, respectively, via the first method. For the second method, the slope value for the Y-axis intercept and R² were -3.321, 31.10 and 0.995, respectively. The PCR inhibitor could not express the detection curve at a copy number over 10¹? via either method, owing to high DNA density. The DNA extract from probiotics was diluted without pre-culturing, and 16 products were amplified via both methods. The Ct value was 11.06~18.12 in the first method and 16.74~22.11 in the second method. Measured probiotics and log copy values were largely similar among the methods used. It was concluded that both methods are effective for analysis, but further research will be required to verify the optimal method.

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