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형광 영상 기반 CD4 신속 검사법 개발을 위한 항원-항체 반응 조건 정량화
김수빈(Subin Kim),김중경(Jung Kyung Kim) 한국가시화정보학회 2015 한국가시화정보학회지 Vol.13 No.1
CD4+ T-cell count determines the effectiveness for antiretroviral therapy (ART) in patients with human immunodeficiency virus (HIV). Although ART slows the progression of HIV to AIDS, rapid counting of CD4+ T lymphocytes with a drop of patient’s blood sample is urgently needed to ensure timely ART treatment in rural areas. Recently point-of-care CD4 testing devices have been developed by using non-flow based imaging cytometer incorporated with a sample cartridge where CD4+ T cells are reacted with fluorescently tagged specific antibodies. Here we conducted an experimental study using a conventional fluorescence microscope-based imaging system to quantitate the interaction of CD4 antibodies with CD4+ T cells at different reaction conditions. We demonstrated that a fast and affordable point-of-care CD4 test is feasible with a far less amount of antibodies and a shorter incubation time compared with a conventional sample preparation protocol for flow cytometry. We also proposed a general method to evaluate and compare the detection limit across different CD4 counting platforms by using fluorescently labelled microbeads for intensity calibration.
자궁 경부암의 임상 조직학적 요인들과 T 조절 림프구 분포와의 상관 분석
권용순 ( Yong Soon Kwon ),김용만 ( Yong Man Kim ),이하영 ( Ha Young Lee ),이신화 ( Shin Wha Lee ),이정남 ( Jung Nam Lee ),김종혁 ( Jong Hyeok Kim ),김영탁 ( Young Tak Kim ),남주현 ( Joo Hyun Nam ) 대한산부인과학회 2007 Obstetrics & Gynecology Science Vol.50 No.12
목적: 자궁 경부암과 T 조절 림프구 특이 Forkhead Box Transcription Factor FoxP3과의 상관 분석을 통해 T 조절 림프구 특이 Forkhead Box Transcription Factor FoxP3의 임상적 의의를 찾고자 한다. 방법: 17명의 자궁 경부암 환자의 말초 혈액을 항 CD4, CD25 mAb 그리고 항 Foxp3 mAb와 결합하는 림프구 세포를 획득하여, flow cytometry을 이용한 Foxp3를 정량화 한다. 이후 자궁 경부암의 임상적, 인구 통계학적 특성과의 상관 분석을 실시한다. 분석된 특성들은 연령, 신장, 체중, 비만도, 최대 종양 직경, 자궁 방 침범 유무, 임파절 침범 유무, 그리고 림프관 종양 침범 유무 등으로 이루어진다. 결과: 여러 자궁경부암의 임상적, 인구 통계학적 특성들 중 T 조절 림프구 특이 Forkhead Box Transcription Factor FoxP3는 최대 종양 직경과 통계적 유의한 상관성을 갖는다 (p value = 0.012, Spearman`s rho 상관 계수 = 0.593). 결론: 진행된 자궁 경부암의 초기 종양의 크기와 말초 혈액의 T 조절 림프구 특이 Forkhead Box Transcription Factor FoxP3사이에는 통계적 유의한 상관관계를 나타내며, 이는 종양 크기와 항 종양 면역 체계와의 유의한 상관성을 지님을 시사한다. Objective: The aim of this study is to determine which clinicopathological factors have the significance in proportion of regulatory T lymphocytes in patients with cervical cancers. Methods: Blood samples were collected preoperatively from 17 patients with uterine cervical cancers (stage: IB, IIA) diagnosed at Asan medical center from May 2006 to October 2006. Patients were limited to who never been treated after diagnosis. We analyzed phenotypes of lymphocytes through flow cytometry by using anti CD3 antibodies, anti CD4 antibodies, anti CD25 antibodies and anti Transcription Factor Foekhead Box P3 (FoxP3) antibodies. Following analysis by flow cytometry, it was determined the relation between the proportion of regulatory T lymphocytes and clinicopathological factors of patients, including age, height, weight, BMI, maximum tumor diameter, lymphovascular tumor emboli, parametrial invasion, and lymph node metastasis. Results: Among many clinicopathological factors in patients with cervical cancer, only maximum tumor diameter was significantly correlated with the proportion of CD4+CD25+highFoxP3+ regulatory T lymphocytes (p=0.012, Spearman`s rho Correlation Coefficient=0.593). Conclusion: This is the first report to document the relation between uterine cervical cancer and the proportion of regulatory T lymphocytes in peripheral blood. In terms of positive correlation between the primary tumor size and the proportion of regulatory T lymphocytes in peripheral blood, we suggest that the spread mechanism of uterine cervical caner, especially direct invasion, will be related with the anti-tumor immune tolerance state. Further studies are necessary to explain the specific spread mechanisms and systemic antitumor immunity in cervical cancer.
인체면역결핍바이러스 감염 환자에서 발생한 이비인후과 질환의 분석
조규섭,이덕규,박영대,신성찬,이선희,노환중 대한이비인후과학회 2009 대한이비인후과학회지 두경부외과학 Vol.52 No.6
Background and Objectives:As human immunodeficiency virus (HIV) infection increases in prevalence and new medical therapies prolong the survival of HIV-infected patients, otolaryngologists will see a larger number of patients with head and neck manifestations of HIV infection. The aims of study were to evaluate the various otorhinolaryngologic diseases and to analyze the useful markers for predicting immune function in HIV-infected patients. Subjects and Method:Of 380 HIV-infected patients, 40 patients (10.5%;39 males and 1 female, age ranging from 23 to 55) were referred to otorhinolaryngologists. Spectrum of disease and correlation between various surrogate markers (HIV RNA, leukocyte count, neutrophil, lymphocyte, CD3, CD4, CD8 T cell percentage and count, CD4/CD8 ratio) and risk of infectious otorhinolaryngologic diseases were analyzed. Results: Thirty-three patients (82.5%) were infectious, 6 patients (15.0%) had non-infectious diseases and 1 patient (2.5%) malignancy. In otologic diseases, acute otitis media and chronic otitis media were frequently found in the mentioned order. In rhinologic diseases, rhinosinusitis and vestibulitis were frequently found in the mentioned order, with one primary extra nodal non-Hodgkin’s lymphoma. In laryngologic diseases, pharyngolaryngitis, oral candidiasis, stomatitis were frequently found. In the surrogate markers of immune function, CD4 percentage, CD4 count and CD4/CD8 ratio were found significantly lower in the infectious diseases than in the non-infectious disease. Conclusion:Of HIV-infected patients, 10.5% had seen an otolaryngologist, and the majority had infectious diseases. The risk of infectious diseases was strongly correlated with the CD4 percentage, count and CD4/CD8 ratio.
회전하는 나선형 마이크로 채널을 이용한 입자 분리 및 정렬
김수빈(Subin Kim),비빈프라사드(Bibin Prasad),김중경(Jung Kyung Kim) 한국가시화정보학회 2015 한국가시화정보학회 학술발표대회 논문집 Vol.2015 No.12
Separation of particles based on different sizes, detection of pathogenic bacteria and isolation of leukocytes from whole blood are typical applications of spiral or helical microchannels. Several microfluidic point-of-care (POC) devices have been developed for monitoring HIV/AIDS by counting CD4 cells but researches are still pursuing to develop a more efficient low-cost device that can be used in developing and underdeveloped countries. The present study focuses on developing a CD4+ T-cell counting device for HIV/AIDS monitoring with the aid of a helical microchannel. For the experiment 10 μm sized particles were used for visualization with a fluorescence microscope system. A sample with the viscosity as that of blood and other samples with different viscosities were also prepared to determine the effect of density and viscosity on alignment of the particles. The channels were spun using a DC motor controlled by an Arduino board with a Bluetooth shield. It was found that for low viscosity samples the particles were not aligned when the channel was kept stationary and an alignment was achieved when the channel was spun due to the combination of centrifugal and gravitational forces. For a sample with viscosity as that of blood, alignment of particles were obtained even without spinning. Since an alignment of particles was achieved for a sample with viscosity as that of blood, the same approach can be applied for aligning and counting CD4+ T cells in blood samples collected from patients.