• Title/Summary/Keyword: flow cytometry

Search Result 1,225, Processing Time 0.025 seconds

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

  • Choi, Woo Young;Park, Cheol;Kim, Gi Young;Lee, Won Ho;Bae, Song-Ja;Choi, Yung Hyun
    • Journal of Marine Bioscience and Biotechnology
    • /
    • v.1 no.2
    • /
    • pp.76-83
    • /
    • 2006
  • Epidemiological studies have indicated that the ubiquitous consumption of seaweeds is a protective factor against some types of cancer. Previous results showed that the administration of seaweed powder or extract reduced the incidence rate of chemically induced tumorigenesis using in vivo animal model. Recently, we reported that the extracts of Gloiopeltis furcata, a kind of Korean edible seaweed, caused he cell growth inhibition of various human cancer cell lines, among them methanol extract exhibited a relatively strong antiproliferative activity. However, the molecular mechanisms of this seaweed in malignant cells have been poorly studied until now. To elucidate this problem, we investigated the effects of methanol extract of G. furcata (MEGF) on the growth inhibition in several human cancer cell lines, and further we analyzed the effects of this extract were tested on the activity of apoptosis induction in human leukemic cells. The results demonstrated that MEGF treatment resulted in the morphological changes and the growth inhibition in a dose-dependent manner. Furthermore, MEGF potently suppresses the growth of human leukemic U937 cells by induction of apoptosis, which was associated with induction of cyclin-dependent kinase inhibitor p21(WAF1/CIP1) in a tumor suppressor p53-independent fashion and up-regulation of Fas/FasL system. Further studies will be needed to identify the active compounds that confer the anticancer activity of MEGF. Once such compounds are identified, the mechanisms by which they exert their effects can begin to be characterized.

  • PDF

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
    • /
    • v.11 no.6
    • /
    • pp.470-478
    • /
    • 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.

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

  • Kang, Joon Won;Quan, Zhejiu;Jang, Jiho;Kang, Hoon-Chul
    • Journal of the Korean Child Neurology Society
    • /
    • v.23 no.2
    • /
    • pp.45-50
    • /
    • 2015
  • Purpose: X-linked adrenoleukodystrophy (X-ALD) is a fatal, axonal demyelinating, neurodegenerative disease, and is caused by mutations the in ABCD1 (ATP-binding cassette transporter subfamily D member 1). Oxidative damage of proteins caused by very long chain fatty acid accumulating in X-ALD, is an early event in the neurodegenerative cascade. We evaluated valproic acid (VPA) as a possible option for oxidative damage in X-ALD. Method: We generated fibroblast of the childhood cerebral ALD from patient. We evaluated mRNA (ribonucleic acid) level of ABCD2 by real-time polymerase chain reaction, and reactive oxygen species (ROS) levels by flow cytometry. Results: VPA increased expression of ABCD2 in both control and ALD fibroblast. ABCD2 gene mRNA expression was increased 1.76 fold in normal fibroblasts, and 2.22 fold in the X-ALD fibroblasts. ROS levels were decreased in VPA treated X-ALD fibroblast, especially in treated with 1 mM of VPA. ROS levels revealed 13.7 in control fibroblast, on the other hand, 5.83 in X-ALD fibroblast treated with 1 mM of VPA. Conclusion: We propose VPA as a promising novel therapeutic approach in oxidant damage that warrants further clinical investigation in X-ALD.

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
    • /
    • v.10 no.6
    • /
    • pp.698-715
    • /
    • 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
    • /
    • v.44 no.3
    • /
    • pp.152-158
    • /
    • 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
    • /
    • v.39 no.1
    • /
    • pp.22-28
    • /
    • 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.

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

  • Baek, Jin Ee;Shin, Jae Hoon;Hwang, Joo Hwan;Lee, Youlim;Lee, Jong Seong;Choi, Byung-Soon
    • Journal of Korean Society of Occupational and Environmental Hygiene
    • /
    • v.31 no.3
    • /
    • pp.286-293
    • /
    • 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.

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

  • Won, JongSeok;Lee, Yeonjung;Lee, Howon;Noh, Jae Hoon
    • Ocean and Polar Research
    • /
    • v.43 no.4
    • /
    • pp.279-293
    • /
    • 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.

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

  • Koo, Bon-Kyeong
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.51 no.1
    • /
    • pp.1-14
    • /
    • 2019
  • The names of medical technologist in Korea, Japan, and the United State of America vary depending on the medical system, educational system, and work areas. Conventionally, jobs equal or similar to medical technologist (commonly known as clinical laboratory technologist, medical laboratory technologist, clinical laboratory scientist, and medical laboratory scientist), histotechnologist, cytotechnologist, and genetic technologist. Jobs, such as extracorporeal technologist, medical electrophysiology technologist, and medical sonographer, allow other allied health professionals including medical technologists, to obtain their qualification. The work areas of medical technologists are classified into clinical pathologic technology, pathologic technology, clinical genetics technology, nuclear medicine in vitro technology, and clinical physiologic technology. New titles of Korean specialized medical technologists refer to certification systems, such as CLMJ, CMAJ, ASCP, IAC, ABRET, and ARMDS. Currently, there are eight types of specialized medical technologists under the cooperation of specialized academic societies. They are hematology, transfusionology, chemistry, immunology, microbiology, histology, cytology, nuclear medicine. It can be further divided into 20 categories according to the work areas such as mass spectrometry, infection control, Pathologists' Assistant, flow cytometry, HLA, cytogenetics, molecular genetics, cardiopulmonary physiology, neurophysiology (EEG/EMG/PSG/etc.), cardiac sonography, neurosonography (TCD/carotid), and medical device quality manager.

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

  • Park, Jae-Eun;Lee, Jun-Young;Lee, Min-Woo;Jang, Eun-Jin;Hong, Chang-Oh;Kim, Keun Ki
    • Journal of Life Science
    • /
    • v.28 no.11
    • /
    • pp.1321-1331
    • /
    • 2018
  • The aim of this study was to separate the photosensitizer that induces apoptosis of U937 and SK-HEP-1 cells from Nostoc commune. Dried N. commune was extracted with $CH_2Cl_2/MeOH$ (1:1) to separate the photosensitizer using various chromatographic techniques. The isolated compound was identified as pheophytin a ($C_{55}H_{74}N_4O_5$) with a molecular weight of 870. Its photodynamic activities were assessed under different irradiation conditions (light and non-light) at the same concentration range of $1.15-23.0{\mu}M$. The apoptosis inducing activity in U937 or SK-HEP-1 cells appeared only in the light. The mechanisms underlying the pheophytin a-mediated photodynamic inhibition of cancer cells were further investigated by examining cell morphology changes, cytotoxicity, caspase-3/7 activity, fluorescence staining, flow cytometry analysis, and DNA fragmentation in these two cell lines. The positive control and the light irradiation group showed typical apoptotic responses, including morphological changes, cytotoxicity, caspase activity, nucleus shrinkage owing to chromatin condensation, DNA laddering, and the presence of apoptotic bodies. Cytotoxicity markedly increased in a dose-dependent manner after a 12 hr exposure. Caspase-3/7 activity was higher in U937 cells than in SK-HEP-1 cells. Apoptosis induction therefore appeared to be both concentration- and light-dependent. In conclusion, pheophytin a, isolated from the blue green alga N. commune, had a photodynamic apoptosis-inducing effect on U937 and SK-HEP-1 cells. The findings reported here can be used as basic data for the development of next-generation photosensitizers from N. commune.