• Title/Summary/Keyword: In vivo flow cytometer

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In vivo Imaging Flow Cytometer (세포 이미징 기능을 겸비한 생체 유세포 분석기)

  • Lee, Ho
    • Journal of the Korean Society of Visualization
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    • v.5 no.1
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    • pp.9-11
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    • 2007
  • We introduce an in vivo imaging flow cytometer, which provides fluorescence images simultaneously with quantitative information on the cell population of interest in a live animal. As fluorescent cells pass through the slit of light focused across a blood vessel, the excited fluorescence is confocally detected. This cell signal triggers a strobe beam and a high sensitivity CCD camera that captures a snap-shot image of the cell as it moves down-stream from the slit. We demonstrate that the majority of signal peaks detected in the in vivo flow cytometer arise from individual cells. The instrument's capability to image circulating T cells and measure their speed in the blood vessel in real time in vivo is demonstrated. The cell signal irradiance variation, clustering percentage, and potential applications in biology and medicine are discussed.

Cell Cycle Arrest by Treatment of D-Ala2-Leu5-enkephalin in Human Leukemia Cancer U937 Cell. (인체혈구암세포 U937의 D-Ala2-Leu5-enkephalin처리에 의한 세포 주기 억제 효과)

  • Lee, Jun-Hyuk;Choi, Woo-Young;Choi, Yung-Hyun;Choi, Byung-Tae
    • Journal of Life Science
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    • v.19 no.5
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    • pp.620-624
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    • 2009
  • D-Ala2-Leu5-enkephalin (DADLE), a hibernation inducer, can induce hibernation-like state in vivo and in vitro. We treated U937 human leukemia cancer cells with DADLE and investigated its possible effect on transcription and proliferation. Treatment of U937 cells with DADLE resulted in growth inhibition and induction of apoptotic cell death on high-dose as measured by MTT assay and DNA flow cytometer analysis. Bcl-XL, c-IAP-2 and survivin genes especially showed decreases in mRNA levels. DADLE treatment also inhibited the levels of cyclooxygenase (COX)-2 mRNA without alteration of COX-1 expression. DNA flow cytometer analysis revealed that DADLE caused arrest of the cell cycle on low-dose, which was associated with a down-regulation of cyclin E at the transcriptional level. DADLE treatment induced a marked down-regulation of cyclin-dependent kinase (Cdk)-2, -4 and -6. In addition, treatment with DADLE decreased telomere associated genes such as, c-myc and TERT, and increased TEP-1 in U937 cells. These results suggest that DADLE can be an inhibition agent in the cell cycle of the human leukemia cancer U937 cell.

The Screening Condition for the Immune Regulatory Responsor Using Mouse Fetal Thymic Organ Culture (쥐의 태아 흉선 조직 배양을 이용한 면역조절제 검색방법 확립)

  • Lee, Seung-Gak;Song, Min-Dong;Lee, Kwang-Ho
    • Korean Journal of Pharmacognosy
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    • v.28 no.4
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    • pp.286-292
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    • 1997
  • We studied the screening condition for immune regulatory responsor. We focused on the T-lymphocytes leer this purpose. Mouse fetal thymic organ culture (FTOC) system and flow cytometric analysis were mainly used in this experiment. Even if FTOC is carried out in vivo condition, the pattern of thymic development in the condition of FTOC is similar to that of in vivo condition. In this regard, FTOC system might be very powerful tool to screen the immune regulator, especially concerning on T cells. To establish the optimum condition of FTOC to screen the Immune regulator, we focused on the optimum amount of dose and culture period. The cell number and surface antigens on T cells were also analysed by using hemacytometer and flow cytometer. To monitor the differentiation event, anti-CD3, anti-CD4 and anti-CD8 antibodies were used. Alkoxyglycerol and Phellodendri Cortex were used fur positive and negative control, respectively. Astragalus membranceus was used as test sample. From our analysis, we reached to conclusions that the best dose of extract is $50\;{\mu}g/ml$ of culture medium, the best culture period is for 9 days, and ethanol used as solvent has no toxicity to FTOC.

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Effect of Gleditsiae Spina extract on immune cells and cytokines in OVA-induced asthmatic mice (급각자가 천식모델 생쥐의 면역세포 및 사이토카인에 미치는 영향)

  • Song, Sang-Jin;Park, Yang-Chun
    • The Journal of Internal Korean Medicine
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    • v.26 no.1
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    • pp.143-155
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    • 2005
  • Objective : The purpose of this research is to examine the effects of Gleditsiae Spina (GS) extract on immune cells and cytokines in ovalbumin (OVA)-induced asthmatic mice. Methods : In vivo, C57BL/6 mice were sensitized and handicapped by OVA for 12 weeks. During this experiment, the one group was then treated with GS extract for the later 8 weeks (3 times per week) and analyzed by ELISA, flow cytometer and RT-PCR. Results : In vivo, there were significant decreases in eosinophils, IL-4, IL-5, IL-13, IgE in BALF (bronchoalveolar lavage fluid). However, $IFN-{\gamma}$ in BALF of GS group increased significantly, compared with that of control group. Additionally, the population of $CD3e^-/CCR3^+,\;CD69^+/CD3e^+,\;IgE^+/B220^+,\;CD11b^+/Gr-l^+$ cells in the GS group decreased. Conclusion : The results of this study support a role for GS as an effective treatment for asthma in its experimental success in significantly decreasing inflammation and asthma reactions, and in increasing $IFN-{\gamma}$, which helps prevent such reactions.

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Effects of Pinelliae Rhizoma on immunocyte and cytokine production in asthma model mouse (반하(半夏)가 천식모델 생쥐의 면역세포 및 사이토카인에 미치는 영향)

  • Song, Jae-Jin;Park, Yang-Chun
    • The Journal of Internal Korean Medicine
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    • v.26 no.1
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    • pp.156-168
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    • 2005
  • Objective : The purpose of this research is to examine the effects of Pinelliae Rhizoma (PR) extract on immune cells and cytokines in ovalbumin (OVA)-induced asthmatic mice. Methods : In vivo, C57BL/6 mice were sensitized and handicapped by OVA for 12 weeks. During this experiment, the one group was then treated with PR extract for the 8 weeks (3 times per week) and analyzed by ELISA and flow cytometer. Results : In vivo, there were significant decreases in eosinophils, IL-4, IL-5, IL-13, IgE in BALF (bronchoalveolar lavage fluid) compared with that of control group. However, $IFN-{\gamma}$ in BALF of GS group increased significantly, compared with that of control group. Additionally, the population of $CD3e^-/CCR3^+,\;CD69^+/CD3e^+,\;IgE^+/B220^+,\;CD11b^+/Gr-l^+$ cells in the PR group decreased, compared with that of control group. Conclusion : The results of this study support a role for PR as an effective treatment for asthma in its experimental success in significantly decreasing inflammation and asthma reactions, and in increasing $IFN-{\gamma}$, which helps prevent such reactions.

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Effect of Hominis Placenta Herbal Acupuncture on immune cells and cytokines in OVA-induced asthmatic mice (자하거(紫河車) 약침이 천식모델 생쥐의 면역세포 및 사이토카인에 미치는 영향)

  • Lim, Ji-Taek;Park, Yang-Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.2
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    • pp.446-451
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    • 2005
  • This experiment was designed to investigate the effect of Mominis Placenta herbal acupuncture solution(HP-HAS) on immune cells and cytokines in murine asthma model. In vivo C57BL/6 mice were sensitized and challenged with OVA for 12 weeks. The experimental group was treated with Hominis Placenta herbal acupuncture solution(HP-HAS) at P'yesu(BL13) for the later 8 weeks(3 times a week) and analyzed by ELISA, flow cytometer. The results were obtained as follows Eosinophils in BALF(bronchoalveolar lavage fluid) of HP-HAS group decreased significantly compared with that of control group. IL-4, IL-5, IL-13, IgE in BALF of HP-HAS group decreased significantly compared with that of control group. Number of $CD3e^-/CCR3^+$, $CD69^+/CD3e^+$, $CD11b^+/Gr-1^+$ cells in the HP-HAS group decreased compared with that of control group.

Effect of Scutellaria Radix extract drug on immune cells and cytokines in BALF of OVA-induced asthmatic mice (황금이 천식모델 생쥐의 면역세포 및 사이토카인에 미치는 영향)

  • Cho, Cheol-Jun;Lim, Do-Hee;Hwang, Ji-Ho;Yang, Soo-Young;Park, Yang-Chun
    • The Journal of Internal Korean Medicine
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    • v.27 no.1
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    • pp.114-125
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    • 2006
  • objective : The purpose of this research is to examine the effects of Scutellaria Radix(SR) extract on immune cells and cytokines in ovalbumin(OVA)-induced asthmatic mice. Methods : In vivo, C57BL/6 mice were sensitized and handicapped by OVA for 9 weeks. In this experiment, one group was not treated, and the other group was treated with SR extract for six weeks(five times per week) and analyzed by ELISA and flow cytometer. Results : In vivo, there were significant decreases in eosinophils, IL-4, IL-13 in BALF (bronchoalveolar lavage fluid) compared with that of control group. However,$IFN{\gamma}$ in the SR group increased significantly compared with that of control group. Additionally, the population of $CD3e^-/CCR3^+,\;CD69^+/CD3e^+,\;IgE^+/B220^+,\;CD11b^+/Gr-1^+$ cells in the SR group decreased compared with that of control group. conclusion : The results of this study support a role for SR as an effective treatment for asthma in its experimental success in significantly decreasing inflammation and asthma reactions, and in increasing $IFN{\gamma}$, Which helps prevent such reactions.

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Induced Death of Escherichia coli Encapsulated in a Hollow Fiber Membrane as Observed In Vitro or After Subcutaneous Implantation

  • Granicka, L. H.;Zolnierowicz, J.;Wasilewska, D.;Werynski, A.;Kawiak, J.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.224-228
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    • 2010
  • The encapsulation of bacteria may be used to harness them for longer periods of time in order to make them viable, whereas antibiotic treatment would result in controlled release of therapeutic molecules. Encapsulated Escherichia coli GFP (green fluorescent protein) (E. coli GFP) was used here as a model for therapeutic substance - GFP fragments release (model of bioactive substances). Our aim was to evaluate the performance of bacteria encapsulated in hollow fibers (HFs) treated with antibiotic for induction of cell death. The polypropylene-surface-modified HFs were applied for E. coli encapsulation. The encapsulated bacteria were treated with tetracycline in vitro or in vivo during subcutaneous implantation into mice. The HF content was evaluated in a flow cytometer, to assess the bacteria cell membrane permeability changes induced by tetracycline treatment. It was observed that the applied membranes prevented release of bacteria through the HF wall. The E. coli GFP culture encapsulated in HF in vitro proved the tetracycline impact on bacteria viability and allows the recognition of the sequence of events within the process of bacteria death. Treatment of the SCID mice with tetracycline for 8 h proved the tetracycline impact on bacteria viability in vivo, raising the necrotic bacteria-releasing GFP fragments. It was concluded that the bacteria may be safely enclosed within the HF at the site of implantation, and when the animal is treated with antibiotic, bacteria may act as a local source of fragments of proteins expressed in the bacteria, a hypothetical bioactive factor for the host eukaryotic organism.

Effects of Siegesbeckiae Herba on rheumatoid arthritis induced by collagen in MRL/MpJ-Ipr-Ipr Mice (희첨이 RA 병태 모델에서 관련인자 및 조직학적 변화에 미치는 영향)

  • Jo Young Ju;Kim Sung Hoon;Park Jong Ho;Kim Dong Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.403-411
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    • 2003
  • For evaluating the effect of Siegesbeckiae Herba (here after abbreviated as SBH) on rheumatoid arthritis, the experiment was carried out; Incidence of rheumatoid arthritis, IRA(indices of rheumatoid arthritis), immunophenotypes by flow cytometer and histopathological changes in MRL/MpJ-Ipr-Ipr Mice in vivo were studied. The results were obtained as follows: 1. Incidence of RA in MRL/Ipr mice was suppressed to 60% of control by SBH. 2. IRA was significantly reduced for IgG3 and IgM at 20 weeks of age and for IgG2b at 12 and 20 weeks of age in mice by SBH compared with control. 3. Immunophenotypes such as CD4/sup +//CD25/sup +/, CD8/sup +//CD3e/sup +/, CD69/sup +//B220/sup +/, NK/sup +//CD3e/sup +/ were significantly increased by SBH compared with control. 4. In histopathological analysis, SBH suppressed the progression of PMN(polymorphonuciear leukocyte), leukocyte and fibroblast infiltration, and subsynovial soft tissue edema frequently showing in the early stage of the arthritis, and also effectively reduced the degeneration of cartilages and degenerative bone symptoms. These results suggest Siegesbeckiae Herba can be effectively applied to the treatment of rheumatoid arthritis and it is still necessary to isolate effective compound from Siegesbeckiae Herba in the near future.

Fermented red ginseng and ginsenoside Rd alleviate ovalbumin-induced allergic rhinitis in mice by suppressing IgE, interleukin-4, and interleukin-5 expression

  • Kim, Hye In;Kim, Jeon-Kyung;Kim, Jae-Young;Han, Myung Joo;Kim, Dong-Hyun
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.635-644
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    • 2019
  • Background: To increase the pharmacological effects of red ginseng (RG, the steamed root of Panax ginseng Meyer), RG products modified by heat process or fermentation have been developed. However, the antiallergic effects of RG and modified/fermented RG have not been simultaneously examined. Therefore, we examined the allergic rhinitis (AR)-inhibitory effects of water-extracted RG (wRG), 50% ethanol-extracted RG (eRG), and bifidobacteria-fermented eRG (fRG) in vivo. Methods: RBL-2H3 cells were stimulated with phorbol 12-myristate-13-acetate/A23187. Mice with AR were prepared by treatment with ovalbumin. Allergic markers IgE, tumor necrosis factor-${\alpha}$, interleukin (IL)-4, and IL-5 were assayed in the blood, bronchoalveolar lavage fluid, nasal mucosa, and colon using enzyme-linked immunosorbent assay. Mast cells, eosinophils, and Th2 cell populations were assayed using a flow cytometer. Results: RG products potently inhibited IL-4 expression in phorbol 12-myristate-13-acetate/A23187-stimulated RBL-2H3 cells. Of tested RG products, fRG most potently inhibited IL-4 expression. RG products also alleviated ovalbumin-induced AR in mice. Of these, fRG most potently reduced nasal allergy symptoms and blood IgE levels. fRG treatment also reduced IL-4 and IL-5 levels in bronchoalveolar lavage fluid, nasal mucosa, and reduced mast cells, eosinophils, and Th2 cell populations. Furthermore, treatment with fRG reduced IL-4, IL-5, and IL-13 levels in the colon and restored ovalbumin-suppressed Bacteroidetes and Actinobacteria populations and ovalbumin-induced Firmicutes population in gut microbiota. Treatment with ginsenoside Rd significantly alleviated ovalbumin-induced AR in mice. Conclusion: fRG and ginsenoside Rd may alleviate AR by suppressing IgE, IL-4, IL-5, and IL-13 expression and restoring the composition of gut microbiota.