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환자 피부 비늘 및 임상주에서 이중 중합효소 연쇄반응(Nested PCR)을 이용한 Malassezia 균종의 동정
임세웅 ( Se Woong Lim ),신명근 ( Myung Geun Shin ),임주영 ( Ju Young Lim ),윤숙정 ( Sook Jung Yun ),김성진 ( Seong Jin Kim ),이승철 ( Seung Chul Lee ),원영호 ( Young Ho Won ),이지범 ( Jee Bum Lee ) 대한피부과학회 2008 대한피부과학회지 Vol.46 No.4
Background: Due to recent identification of new Malassezia (M.) species, M. dermatis, and M. equi, the genus Malassezia was revised into eleven species that have been isolated from human and animal skin. This has further substantiated the need for molecular techniques to distinguish the various Malassezia species. Objective: We aimed to make the nested polymerase chain reaction (PCR) using species-specific primers with specificity and sensitivity as a diagnostic tool for differentiating the various Malassezia species from skin scales and fungal cells rapidly and accurately. In addition, we evaluated the common causative Malassezia species in the patients with seborreheic dermatitis, pityriasis versicolor or pityrisporum folliculitis. Methods: Malassezia species-specific primers were designed based on DNA sequencing of the ribosomal RNA gene. The standard strains of eight members of the genus Malassezia such M. pachydermatis, M. furfur, M. sympodialis, M. globosa, M. obtuse, M. restricta, M. slooffiae, and M. dermatis were used for positive control. Each Malassezia species was cultured separately and two or three standard species were cultured together on Modified Leeming and Notman agar (MLNA) media plates. In addition, twenty-five clinical strains of Malassezia species isolated from the skin of patients with dermatological conditions and twenty-three samples of skin scale were used as well. Results: The nested PCR assay with primers for all eight Malassezia species were species-specific since it amplified DNA only from the target Malassezia species, and could differentiate mixed, that is, the two or three Malassezia species of all standard strains grown on MLNA medium precisely. Detection of Malassezia species from clinical strains and patient skin scales using the nested PCR assay was 96% (24/25) and 87% (20/23), respectively. M. globosa, M. sympodialis, M. restricta were the most common causative Malassezia species in patients with pityriasis versicolor, pityrosporum folliculitis, seborrheic dermatitis, respectively. Conclusion: Nested PCR using species-specific primers is useful and reliable in the detection of various Malassezia species from patient skin scales as well as cultured fungal cells. (Korean J Dermatol 2008;46(4):446~452)
Random amplified polymorphic DNA 분석에 의한 Helicobacter pylori의 분자적 형별분석
이숭(Soong Lee),신명근(Myung Geun Shin) 대한내과학회 2001 대한내과학회지 Vol.60 No.2
Background : Genetic diversity among Helicobacter pylori strains isolated from different patients has been debated. This study was undertaken to determine molecular types and genetic diversity among 20 isolates of H. pylori obtained from various gastroduodenal diseases, and also examined the association between molecular types of H. pylori and these diseases. Methods : Antral biopsies were taken for culture from 38 patients with chronic gastritis, gastric ulcer and duodenal ulcer at the time of endoscopy. Biopsy specimens were primarily inoculated on chocolate agar and incubated microaerophilically at 37℃ for up to 7 days. H. pylori was identified by typical Gram stain morphology and biochemical tests. Random amplified polymorphic DNA (RAPD) fingerprinting was performed by 4 primers (OPA-07, 5'-GAAACGGGTG-3'; OPA-10, 5'-GTGATCGCAG-3'; OPA-11, 5'-CAATCGCCGT-3'; OPA-12, 5'-TCGGCGATAG-3'; Operon Technologies, Atlanta, GA). We used the NTSYS-pc (numerical taxonomy system and multivariate analysis system, version 1.50, Applied Biostatistics Inc., CA, USA) program to compose the phenogram for the differentiation of H. pylori strains. Results : Twenty strains (52.6%) of H. pylori were isolated from 38 biopsy specimens. All isolates were divided into five molecular types (I-V) at similarity (S) value of 0.63; 7 strains (35%), 4 strains (20%), 4 strains (20%), 3 strains (15%) and 2 strains (10%) belonged to type II, III, IV, V and I, respectively. The distribution of genetic S value was 0.24 to 0.91 in all isolates, thus the isolates had a wide range of S values. The mean S values of all isolates, type I, II, III, IV and V were 0.69, 0.69, 0.73, 0.75 and 0.65, respectively. There was no specific correlation between molecular types and gastroduodenal diseases. Conclusion : H. pylori isolates had high level of genetic diversity. The RAPD molecular types of H. pylori were not disease-specific since the types were diverse in the isolates from various gastroduodenal diseases.(Korean J Med 60:115-122, 2001)
Klebsiella rhinoscleromatis에 의한 외음부질염 1예
김진각 ( Jin Gak Kim ),신명근 ( Myung Geun Shin ) 대한임상검사과학회 1998 대한임상검사과학회지(KJCLS) Vol.30 No.3
A 6-year-old girl presented with foul-odored vaginal discharge of 1 month duration. She was born of uncomplicated first full-term gestation, and the birth weight was 3,500g. Examination disclosed no abnormality. We repeatedly isolated Klebsiella rhinoscleromatis from vaginal discharge during a week. The isolated organism was gram negative bacilli, grew on blood agar and McConkey agar and identified as Klebsiella rhinosclerarotis by using AlR32GN (API system, La Balrre-Les Grottes, France). By conventional biochemical tests, glucose was fermented with the production of acid and gas. Methyl red and malonate utilization were positive, while Voges-Prouskauer (VP) reaction, citrate utilization, urease, phenylalanine deaminase, lysine decarboxylase, arginine dihydrolase, ornithine decarboxylase, motility and DNase were negative. AII these biochemical findings were compatible with Klebsiella rhinosclerarotis. The isolate was susceptible to various antibiotics such as kanamycin, gentamicin, amikacin, tetracycline, chloramphenicol, cephalothin, cefoperazone, ampicillin, carbenicillin. colistin and piperacillin.
김여경,Seung-Shin Lee,안재숙,양덕환,이제정,김형준,Myung Geun Shin,정성훈,신명근 전남대학교 의과학연구소 2014 전남의대학술지 Vol.50 No.3
This study explored drug transporter expression levels and their impact on clinical responseto imatinib and second-generation tyrosine kinase inhibitors (TKIs) in imatinib-resistant chronic myeloid leukemia (CML). Imatinib-resistant chronic phaseCML patients treated with dasatinib (n=10) and nilotinib (n=12) were enrolled. ThemRNA expression of the OCT-1, ABCG2, and ABCB1 genes was quantified by usingpaired bone marrow samples obtained before administering imatinib and at the pointof detecting imatinib resistance (just before starting second-generation TKIs). The expressionlevels of OCT-1 and ABCG2 were lower in follow-up than in imatinib-naïvesamples. ABCB1 revealed highly variable expression levels before and after imatinibtreatment. In addition, median ABCB1 expression in follow-up samples was lower inpatients achieving complete cytogenetic response or major molecular response duringimatinib treatment than in failed patients. Higher ABCG2 expression in imatinib-exposedsamples showed a negative impact on optimal response to dasatinib. Patientswith higher ABCG2 expression in imatinib-exposed samples also had shorter progression-free survival with dasatinib treatment. However, no significant correlationwas found between these drug transporter expression levels in imatinib-naïve or imatinib-exposed samples and responses to nilotinib. In imatinib-resistant CML, OCT-1and ABCG2 mRNA expression decreased after imatinib treatment. Patients with higherABCG2 expression in imatinib-exposed samples showed poor treatment outcomewith dasatinib. On the other hand, a higher expression level of ABCB1 in imatinib-exposedsamples did not affect second-generation TKI responses but was correlated withpoor imatinib responses.
조상혁(Sang Hyuk Cho),허정욱(Jung Wook Huh),김영진(Young Jin Kim),신명근(Myung Geun Shin),김형록(Hyeong Rok Kim) 대한외과학회 2009 Annals of Surgical Treatment and Research(ASRT) Vol.76 No.3
Purpose: APC (adenomatous polyposis coli) gene is one of the tumor-suppressor genes that acts in the early stages of cancer. Among general colon cancer patients, normal APC gene expression is deficient in 80%. It seems that APC is the most important gene in the development of colon cancer. This study was performed to analyze the mutation spectra of APC gene in sporadic colon cancer tissue from Korean patients with colon cancer. Methods: A total of 38 patients with sporadic colon cancer were enrolled. Colon cancer tissues were analyzed for the determination of APC gene mutation spectra by multiplex ligation-dependent probe amplification (MLPA) method using SALSA MLPA P403 APC kit (MRC-Holland, Amsterdam, NL). Results: APC gene mutations showing deletion/duplication in one or more exons were detected in 23 (60.5%) patients. Duplication in 13 patients (56.5%), duplication and deletion in 7 patients (30.4%), and deletion in 3 patients (13.1%) was detected. The incidence of APC gene mutation found in this study was highest in exon 3. From this study, no significant differences were observed with respect to clinicopathologic findings and the presence or absence of APC mutations. Conclusion: The frequency of APC gene mutation was about 61% in Korean patients with colon cancer, it showed concordance with the previous reports on the frequency of APC gene mutation from Caucasian patients with sporadic colon cancer. However, in contrast to these reports, the frequency of duplication disclosed much higher than those of western countries.
급성 골수성 백혈병 환자에서 발생한Fusarium solani에 의한 전신 감염증
임세웅 ( Se Woong Lim ),김형준 ( Hyung Joon Kim ),신명근 ( Myung Geun Shin ),이재혁 ( Jae Hyuk Lee ),윤숙정 ( Sook Jung Yun ),이지범 ( Jee Bum Lee ),김성진 ( Seong Jin Kim ),원영호 ( Young Ho Won ),이승철 ( Seung Chul Lee ) 대한피부과학회 2008 대한피부과학회지 Vol.46 No.6
Fusarium species are known as ubiquitous soil saprophytes and human skin contaminants. Infections caused by Fusarium species are increasing in frequency among immunocompromised hosts. We report a rare case of fatal disseminated infection caused by Fusarium solani in an acute myeloid leukemia patient. Skin biopsy specimen revealed multiple branched hyphae and spores in the vessel with thrombus formation, and fungal culture showed a fuzzy, whitish colony. Direct sequencing of 28S rRNA gene confirmed the isolate as Fusarium solani. (Korean J Dermatol 2008;46(6):846∼850)
논문 1 : 백혈병 미세잔존질환 정량검출을 위한 실시간 역전사중합효소연쇄반응법의 유용성
조정애 ( Jeung Ai Cho ),김다운 ( Da Woon Kim ),정성두 ( Seong Du Jeong ),천지선 ( Ji Seon Cheon ),나경아 ( Gyeong Ah Na ),김진각 ( Jin Gak Kim ),김인환 ( In Hwan Kim ),김수현 ( Soo Hyun Kim ),신명근 ( Myung Geun Shin ) 대한임상병리사협회 2008 임상혈액검사학회 발표자료집 Vol.10 No.1
Background : Chromosomal rearrangements are major pathology inhematological malignancy. The detection of minimal residual disease(MRD) for these gene rearrangements helps in monitoring treatment outcomes and predicting prognosis. patients. Recently, quantification of these gene transcripts based on real-time quantitative polymerase chain reaction (RQ-PCR) has been used as MRD detection. The purpose of this study is to ensure the usefulness of the RQ-PCR technique for detecting MRD in hematological malignancy patients. Methods : The patients had been diagnosed to AML1-ETO positive AML, PML-RARa positive AML and BCR-ABL positive MPN (CML) at Chonnam National University Hwasun Hospital (CNUHH, Hwasun, Korea) from Jan. 2006 to Aug. 2008. The fusion transcript was quantified by RQ-PCR and analyzed in comparison to conventional cytogenetics, FISH and RT-PCR. Results : The fusion gene transcript was quantified by RQ-PCR in 57 samples from 14 patients with AML1-ETO positive AML, 79 samples from 27 patients with PML-RARa positive AML and 108 samples from 36 patients with CML. At diagnosis, the quantitative fusion transcripts for AM1-ETO, PML-RARa and BCR-ABL showed the range of 0.485552651 ~ 10.82233683 (mean 3.782217131, SD 2.998052348), 0.005300395 ~ 0.29267494 (mean 0.056901315, SD 0.080131381) and 0.1293929 ~ 12.94826849 (mean 1.701935665, SD 2.200913158). The increase of AML1-ETO fusion gene transcripts preceded morphologic relapse in two patients. Quantification of fusion gene transcripts by RQ-PCR could detected MRD in samples which were negative by in cytogenetics analysis or FISH. Conclusions : Our findings indicated that quantitative analysis of AML1-ETO, PML-RARa and BCR-ABL transcripts by RQ-PCR might be a useful tool for the monitoring of minimal residual disease in hematological malignancy.