• 제목/요약/키워드: flow cytometry

검색결과 1,225건 처리시간 0.034초

인체백혈병세포의 증식에 미치는 불등가사리 메탄올 추출물의 영향 (Induction of Apoptosis by Methanol Extract of Gloiopeltis furcata in Human Leukemia Cell Line U937)

  • 최우영;;김기영;이원호;배송자;최영현
    • 한국해양바이오학회지
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    • 제1권2호
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    • pp.76-83
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    • 2006
  • 본 연구에서는 다양한 인체암세포의 증식에 미치는 등불가사리 메탄올 추출물(MEGF)의 영향을 조사하였다. MEGF는 처리 농도의존적으로 암세포의 형태적변이 및 증식억제가 효과를 보여주었으며, 특히 백혈병세포에서 가장 높은 감수성을 보여주었다. 따라서 백혈병세포의 증식억제 효과가 apoptosis 유발과 연관성이 있는지를 flow cytometry 분석 및 DAPI staining 법으로 조사한 결과, MEGF 처리에 의한 백혈병세포의 증식억제는 세포주기 교란과 무관한 apoptosis 유발에 의한 것임을 sub-G1기 세포의 빈도 증가 및 apoptotic body 형성의 증가 등으로 확인하였다. 또한 MEGF 처리에 의한 백혈병세포의 증식억제 및 apoptosis 유발은 p53 비의존적인 p21의 발현 증가 및 Fas/FasL system의 발현 증가와 연관성이 있음을 알 수 있었다. 이상의 결과들은 인체 암세포, 특히 백혈병세포에서 MEGF의 항암작용을 이해하는데 중요한 자료가 될 것이고 나아가 MEGF 내 함유된 생리활성 물질의 분리 및 항암적용 연구를 위한 중요한 기초 자료로 활용될 것이다.

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Anti-fibrotic effects of Orostachys japonicus A. Berger (Crassulaceae) on hepatic stellate cells and thioacetamide-induced fibrosis in rats

  • Koppula, Sushruta;Yum, Mun-Jeong;Kim, Jin-Seoub;Shin, Gwang-Mo;Chae, Yun-Jin;Yoon, Tony;Chun, Chi-Su;Lee, Jae-Dong;Song, MinDong
    • Nutrition Research and Practice
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    • 제11권6호
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    • pp.470-478
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    • 2017
  • BACKGROUND/OBJECTIVE: Orostachys japonicus A. Berger (Crassulaceae) has been used in traditional herbal medicines in Korea and other Asian countries to treat various diseases, including liver disorders. In the present study, the anti-fibrotic effects of O. japonicus extract (OJE) in cellular and experimental hepatofibrotic rat models were investigated. MATERIALS/METHODS: An in vitro hepatic stellate cells (HSCs) system was used to estimate cell viability, cell cycle and apoptosis by MTT assay, flow cytometry, and Annexin V-FITC/PI staining techniques, respectively. In addition, thioacetamide (TAA)-induced liver fibrosis was established in Sprague Dawley rats. Briefly, animals were divided into five groups (n = 8): Control, TAA, OJE 10 (TAA with OJE 10 mg/kg), OJE 100 (TAA with OJE 100 mg/kg) and silymarin (TAA with Silymarin 50 mg/kg). Fibrosis was induced by treatment with TAA (200 mg/kg, i.p.) twice per week for 13 weeks, while OJE and silymarin were administered orally two times per week from week 7 to 13. The fibrotic related gene expression serum biomarkers glutathione and hydroxyproline were estimated by RT-PCR and spectrophotometry, respectively, using commercial kits. RESULTS: OJE (0.5 and 0.1 mg/ mL) and silymarin (0.05 mg/mL) treatment significantly (P < 0.01 and P < 0.001) induced apoptosis (16.95% and 27.48% for OJE and 25.87% for silymarin, respectively) in HSC-T6 cells when compared with the control group (9.09%). Further, rat primary HSCs showed changes in morphology in response to OJE 0.1 mg/mL treatment. In in vivo studies, OJE (10 and 100 mg/kg) treatment significantly ameliorated TAA-induced alterations in levels of serum biomarkers, fibrotic related gene expression, glutathione, and hydroxyproline (P < 0.05-P < 0.001) and rescued the histopathological changes. CONCLUSIONS: OJE can be developed as a potential agent for the treatment of hepatofibrosis.

부신백질형성장애증 섬유모세포에서 발프로산의 항산화능 (Valproic Acid Reduces Reactive Oxygen Species in Fibroblast of X-linked Adrenoleukodystrophy)

  • 강준원;전철구;장지호;강훈철
    • 대한소아신경학회지
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    • 제23권2호
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    • pp.45-50
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    • 2015
  • 목적: X-ALD는 Xq28에 위치한 ABCD1 유전자의 돌연변이로 긴사슬지방산이 신경 조직과 부신에 축적되어 일어나는 퇴행 뇌질환이며, 소아기 대뇌형의 경우 빠르고 심한 임상 경과를 보인다. 이 과정에서 산화스트레스도 조직 손상에 영향을 주는 것으로 알려져 있다. 골수이식이나 로렌조 기름 등이 치료 방법으로 이용되나 치료의 위험성과 효과에서 한계를 보이는 실정이다. 이에 저자들은 X-ALD 환자에게 채취한 섬유모세포를 이용하여, X-ALD의 치료 가능성을 알아보기 위하여 VPA의 효과를 연구해보고자 하였다. 방법: X-ALD 환자의 피부에서 채취한 섬유모세포와 정상인의 피부에서 채취한 섬유모세포를 배양하였다. 배양된 섬유모세포에 VPA를 처리한 후 RNA발현 정도를 통해 ABCD2 발현을 확인하고 유동세포계측법으로 활성산소종을 측정하였다. 결과: VPA을 처리한 후 정상과 X-ALD 섬유모세포 모두에서 ABCD2의 mRNA 발현이 증가하였다. 특히 X-ALD 섬유모세포에서 ABCD2 유전자 mRNA 발현이 2.22배로 정상의 1.76배보다 더 증가하였다. 유동세포계측법으로 활성산소종을 확인한 결과 대조군에서 13.7, VPA를 처리한 군에서는 각각 0.25 mM에서 8.67, 0.5 mM에서 9.37, 1 mM에서 5.83을 나타내었다. 결론: X-ALD 환자에서 VPA의 항산화능을 이용하여 신경손상을 막을 수 있는 가능성이 있을 것으로 보이며, 이를 실제 환자에 적용하는 연구가 필요할 것이다.

Differential Hrd1 Expression and B-Cell Accumulation in Eosinophilic and Non-eosinophilic Chronic Rhinosinusitis With Nasal Polyps

  • Chen, Kun;Han, Miaomiao;Tang, Mengyao;Xie, Yadong;Lai, Yuting;Hu, Xianting;Zhang, Jia;Yang, Jun;Li, Huabin
    • Allergy, Asthma & Immunology Research
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    • 제10권6호
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    • pp.698-715
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    • 2018
  • Purpose: Hrd1 has recently emerged as a critical regulator of B-cells in autoimmune diseases. However, its role in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) remains largely unexplored. This study aimed to examine Hrd1 expression and B-cell accumulation and their possible roles in CRSwNP. Methods: Quantitative real-time polymerase chain reaction, immunohistochemistry, enzyme-linked immunosorbent assay and Western blotting were used to assess gene and protein expression in nasal tissue extracts. Cells isolated from nasal tissues and peripheral blood mononuclear cells were characterized by flow cytometry. Local antibody production was measured in tissue extracts with a Bio-Plex assay. Additionally, changes in Hrd1 expression in response to specific inflammatory stimuli were measured in cultured dispersed polyp cells. Results: Nasal polyps (NPs) from patients with eosinophilic CRSwNP (ECRS) had increased levels of Hrd1, B-cells and plasma cells compared with NPs from patients with non-eosinophilic CRSwNP (non-ECRS) or other control subjects (P < 0.05). The average Hrd1 levels in B-cells in NPs from ECRS patients were significantly higher than those from non-ECRS patients and control subjects (P < 0.05). NPs also contained significantly increased levels of several antibody isotypes compared with normal controls (P < 0.05). Interestingly, Hrd1 expression in cultured polyp cells from ECRS patients, but not non-ECRS patients, was significantly increased by interleukin-$1{\beta}$, lipopolysaccharide and Poly(I:C) stimulation, and inhibited by dexamethasone treatment (P < 0.05). Conclusions: Differential Hrd1 expression and B-cell accumulation between the ECRS and non-ECRS subsets suggests that they can exhibit distinct pathogenic mechanisms and play important roles in NP.

MTHFR 3'-untranslated region polymorphisms contribute to recurrent pregnancy loss risk and alterations in peripheral natural killer cell proportions

  • Kim, Eun Sun;Kim, Jung Oh;An, Hui Jeong;Sakong, Jung Hyun;Lee, Hyun Ah;Kim, Ji Hyang;Ahn, Eun Hee;Kim, Young Ran;Lee, Woo Sik;Kim, Nam Keun
    • Clinical and Experimental Reproductive Medicine
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    • 제44권3호
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    • pp.152-158
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    • 2017
  • Objective: To identify the associations between polymorphisms of the 3'-untranslated region (UTR) of methylenetetrahydrofolate reductase (MTHFR) gene, which codes for an important regulatory enzyme primarily involved in folate metabolism, and idiopathic recurrent pregnancy loss (RPL) in Korean women. Methods: The study population comprised 369 RPL patients and 228 controls. MTHFR 2572C > A, 4869C > G, 5488C > T, and 6685T > C 3'-UTR polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis or by TaqMan allelic discrimination assays. Natural killer cell proportions were determined by flow cytometry. Results: The MTHFR 2572-5488-6685 (A-C-T) haplotype had an adjusted odds ratio of 0.420 (95% confidence interval, 0.178-0.994; p= 0.048) for RPL. Analysis of variance revealed that MTHFR 4869C > G was associated with altered $CD56^+$ natural killer cell percentages (CC, $17.91%{\pm}8.04%$; CG, $12.67%{\pm}4.64%$; p= 0.024) and folate levels (CC, $12.01{\pm}7.18mg/mL$; CG, $22.15{\pm}26.25mg/mL$; p= 0.006). Conclusion: Variants in the 3'-UTR of MTHFR are potential biomarkers for RPL. However, these results should be validated in additional studies of ethnically diverse groups of patients.

Effect of sun ginseng potentiation on epirubicin and paclitaxel-induced apoptosis in human cervical cancer cells

  • Lin, Yingjia;Jiang, Dan;Li, Yang;Han, Xinye;Yu, Di;Park, Jeong Hill;Jin, Ying-Hua
    • Journal of Ginseng Research
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    • 제39권1호
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    • pp.22-28
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    • 2015
  • Background: Sun ginseng (SG), a specific formulation of quality-controlled red ginseng, contains approximately equal amounts of three major ginsenosides (RK1, Rg3, and Rg5), which reportedly has antitumor-promoting activities in animal models. Methods: MTT assay was used to assess whether SG can potentiate the anticancer activity of epirubicin or paclitaxel in human cervical adenocarcinoma HeLa cells, human colon cancer SW111C cells, and SW480 cells; apoptosis status was analyzed by annexin V-FITC and PI and analyzed by flow cytometry; and apoptosis pathway was studied by analysis of caspase-3, -8, and -9 activation, mitochondrial accumulation of Bax and Bak, and cytochrome c release. Results: SG remarkably enhances cancer cell death induced by epirubicin or paclitaxel in human cervical adenocarcinoma HeLa cells, human colon cancer SW111C cells, and SW480 cells. Results of the mechanism study highlighted the cooperation between SG and epirubicin or paclitaxel in activating caspase-3 and -9 but not caspase-8. Moreover, SG significantly increased the mitochondrial accumulation of both Bax and Bak triggered by epirubicin or paclitaxel as well as the subsequent release of cytochrome c in the targeted cells. Conclusion: SG significantly potentiated the anticancer activities of epirubicin and paclitaxel in a synergistic manner. These effects were associated with the increased mitochondrial accumulation of both Bax and Bak that led to an enhanced cytochrome c release, caspase-9/-3 activation, and apoptosis. Treating cancer cells by combining epirubicin and paclitaxel with SG may prove to be a novel strategy for enhancing the efficacy of the two drug types.

만성폐쇄성폐질환을 동반한 무기분진 노출 이직근로자의 혈중 림프구 아형분포에 미치는 영향요인 (Influencing Factors of Peripheral Blood Lymphocyte subsets in Workers with Chronic Obstructive Pulmonary Disease Exposed to Inorganic Dust)

  • 백진이;신재훈;황주환;이유림;이종성;최병순
    • 한국산업보건학회지
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    • 제31권3호
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    • pp.286-293
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    • 2021
  • Objectives: Inorganic dust is known to be a risk factor for chronic obstructive pulmonary disease (COPD) regardless of smoking and pneumoconiosis. Adaptive and innate immunity, including lymphocyte infiltrate, are involved in the pathogenesis of COPD. The purpose of this study was to analyze the lymphocyte subsets in the blood of workers exposed to inorganic dust and confirm the influencing factors. Methods: The general characteristics of the subjects (n=107) were analyzed through a personal questionnaire. Diagnosis of COPD was established according to pulmonary function tests with FEV1/FVC post bronchodilator lower than 70%, according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) guidelines. For lymphocyte analysis, blood was stained with a fluorescent CD marker and analyzed by flow cytometry. Results: The increase in CD4+ T lymphocytes was associated with a decrease in age (𝛽=-0.273, p=0.008) and an increase in the cumulative smoking amount (𝛽=0.205, p=0.034). The increase in NK cells was associated with an increase in age (𝛽=0.325, p=0.001) and a decrease in cumulative smoking (𝛽=-0.220, p=0.019). The period of exposure to dust, %FVC predicted and %FEV1/FVC, and the relative population of peripheral blood lymphocytes did not show a statistically significant relationship. Conclusions: CD4+ T lymphocytes and CD56+CD16+ NK cells in peripheral blood were more related to age and cumulative smoking than the duration of dust exposure. Age and smoking are major risk factors for the development of COPD, so it can be predicted that peripheral blood CD4+ T lymphocytes and CD56+CD16+ NK cells are related to the development of COPD in workers exposed to inorganic dust.

남해 해역의 해양환경변화가 시아노박테리아 개체수와 군집 조성에 미치는 영향 (Effect of Marine Environment Changes on the Abundance and Community Composition of Cyanobacteria in the South Sea of Korea)

  • 원종석;이연정;이호원;노재훈
    • Ocean and Polar Research
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    • 제43권4호
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    • pp.279-293
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    • 2021
  • To investigate the effect of seasonal marine environment conditions on the cyanobacteria abundance and diversity in the South Sea, four-seasonal surveys were conducted along the 127.5°E survey transect line in the central South Sea using flow cytometry and 16S-23S ITS on the Miseq platform from August 2016 to May 2017. The average abundance of Synechococcus varied from 3.3 × 103 to 7.4 × 104 cells ml-1. The abundance was the highest in the summer and the lowest in the winter, and the abundance fluctuated according to water temperature. The abundance was high in the outer sea affected by TWC. However, in summer, the Coastal areas affected by the Yangtze River were more populated than the outer sea. Prochlorococcus was rare and could not penetrate into coastal areas due to the fronts, but showed its dominance in the waters influenced by the TWC. Synechococcus clades II, VII, IX, CRD1, and CRD2 were predominant in the outer sea area affected by the TWC. In the coastal area, clades I and IV showed higher dominance whereas clades V, VI, WPC1, and 5.3-MS3 with euryhaline characteristics, showed a high dominance rate in the water masses affected by the low-salinity water of the Yangtze River in the summer. Clade XVI, XVII, CB1, CB5, and 5.3-I/II showed high dominance in nutrient-rich waters in the summer with increased water temperature. The abundance and community composition of cyanobacteria changed in the South Sea due to the influence of the TWC and stratification. In the summer, the abundance and the community composition differed, and were mainly affected by the general influence of the TWC in addition to the influence of the Yangtze River low-salinity water.

한국, 일본, 미국 임상병리사 전문자격 (Professional Certification of Medical Technologists in Korea, Japan, and United States of America)

  • 구본경
    • 대한임상검사과학회지
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    • 제51권1호
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    • pp.1-14
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    • 2019
  • 우리나라, 일본, 미국의 임상병리사 명칭은 의료체계, 교육제도, 업무영역에 따라 차이가 있다. 전통적으로 medical technologist (별칭 clinical laboratory technologist, medical laboratory technologist, clinical laboratory scientist, medical laboratory scientist)와 동등하거나 유사한 직종은 histotechnologist, cytotechnologist, genetic technologist이다. Extracorporeal technologist, medical electrophysiology technologist, medical sonographer는 medical technologist를 포함한 다른 보건의료인도 자격을 취득할 수 있는 직종이다. 임상병리사의 업무영역은 임상병리기술학, 병리기술학, 임상유전기술학, 핵의학기술학, 임상생리기술학으로 분류하였다. 새로운 전문임상병리사 종별은 CLMJ, CMAJ, ASCP, IAC, ABRET, ARMDS 등의 자격인정제도를 참고하였다. 현재 전문임상병리사 종별은 전문과목학회의 협조 하에 기존의 8종으로 혈액학, 수혈학, 화학, 면역학, 미생물학, 조직학, 세포학, 핵의학분야가 있으며 앞으로 미세질량분석, 감염관리, Pathologists' Assistant, 유세포분석, HLA, 세포유전학, 분자유전학, 심폐생리학, 신경생리학(뇌파/근전도/수면다원 등), 심장초음파학, 신경초음파학(뇌혈류/경동맥), 기기품질관리사 등 업무영역별로 20종으로 세분화할 수 있다.

Nostoc commune으로부터 백혈병세포와 간암세포에 대한 apoptosis 유도 광과민성물질 pheophytin a의 분리 및 구조동정 (Isolation and Identification of Pheophytin, a Photosensitizer from Nostoc commune that Induces Apoptosis in Leukemia and Cancer Cells)

  • 박재은;이준영;이민우;장은진;홍창오;김근기
    • 생명과학회지
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    • 제28권11호
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    • pp.1321-1331
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    • 2018
  • 본 연구에서 N. commune으로부터 U937과 SK-HEP-1 세포에 대해 apoptosis를 유도하는 광과민성물질을 순수 분리하였다. N. commune을 dichloromethane : Methanol (1:1)로 추출하여 Hexane, EtOAc, MeOH로 분획하였다. 높은 광역학활성을 나타내는 EtOAc 분획물에서 NC-1을 분리하였으며, 분자량 870의 pheophytin a ($C_{55}H_{74}N_4O_5$)인 것으로 구조를 동정하였다. 광역학활성 조사는 NC-1을 $1.15{\mu}M$에서 $23.00{\mu}M$까지 5개의 농도를 처리하여 광을 조사하는 것과 조사하지 않는 조건으로 나누어 평가하였다. U937과 SK-HEP-1 두 세포에서 apoptotic body 형성, cell blebbing 등의 형태적 변화와 세포독성, caspase 활성, 세포핵의 응축, DNA 단편화 등이 광 의존적이며 농도 의존적으로 활성이 증가하였다. 추가적으로 유세포분석을 통한 apoptosis 유도의 정량적 분석 결과에서도 광조건에서 농도가 증가할수록 apoptosis 유도가 증가하는 것을 확인하였다. 이러한 결과를 통해 NC-1의 U937과 SK-HEP-1 세포에 대한 사멸은 광역학적 apoptosis에 의한 것임을 확인하였다. 본 연구 내용을 N. commune으로부터 광감각제 개발의 기초자료로 활용할 수 있을 것이다.