• Title/Summary/Keyword: Fluorescence bead

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The Effect of Bacillus-Fermented Scutellariae Radix Acupuncture Solution on Interleukin Production in Mouse Macrophage Stimulated by Lipopolysaccharide (바실러스균 발효황금약침액이 Lipopolysaccharide로 활성화된 마우스 대식세포의 인터루킨 생성에 미치는 영향)

  • Park, Wan-Su
    • Korean Journal of Acupuncture
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    • v.27 no.2
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    • pp.95-105
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    • 2010
  • Objectives : The purpose of this study is to investigate the effect of Bacillus-fermented Scutellariae Radix acupuncture solution (SB) on interleukin(IL) production in mouse macrophage stimulatedby lipopolysaccaride(LPS). Methods : Productions of interleukins were measured y High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$(multi-analyte profiling beads) technology. To begin with, cell culture supernatant was obtained after treatment with LPS(1 ${\mu}g/mL$) and SB for 24 hour. Then, it was incubated with the antibody-conj${\mu}g$ated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. After incubating for 30 minutes, Strepavidin-conjugated Phycoerythrin(SAPE) was then added. Incubating for another 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Results : The results of the experiment are as follows. SB significantly inhibited the LPS-induced production of IL-3($9.15{\pm}0.35$ pg/mL) by $6.92{\pm}0.05,\;7.21{\pm}0.11,\;6.96{\pm}0.33,\;and\;7.45{\pm}0.74$ pg/mL at the concentration of 25, 50, 100, and 200 ${\mu}g/mL$ in mouse macrophage RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced production of IL-5($7.30{\pm}0.48$ pg/mL) by $6.50{\pm}0.29,\;6.30{\pm}0.25,\;6.30{\pm}0.25,\;and\;5.80{\pm}0.25$ pg/mL at the concentration of 25, 50 100, and 200 ${\mg}g/mL$ in RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced productiion of IL-9($17.26{\pm}0.19$ pg/mL) by $15.01{\pm}0.43$ pg/mL at the concentration of 25 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced productioh of IL-13($187.80{\pm}2.90$ pg/mL) by $152.80{\pm}4.25,\;172.80{\pm}3.97,\;162.10{\pm}6.67,\;and\;165.30{\pm}11.80$ pg/mL at the concentration fo 25, 50, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-17($18.30{\pm}0.95$ pg/mL) by $13.30{\pm}1.25,\;13.80{\pm}1.11,\;13.30{\pm}0.75,\;and\;14.00{\pm}1.08$ pg/mL at the concentration of 25, 50 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-23($43.90{\pm}0.83$ pg/mL by $39.50{\pm}1.26,\;38.00{\pm}1.78,\;and\;39.60{\pm}2.49$ pg/mL at the concentration of 25, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). Conclusions : These results suggest that SB has anti-inflammatory activity related with its inhibition of IL-3, IL-5, IL-13, IL-17, and IL-23 production in macrophages.

A Study on Material Characteristics and Manufacturing Techniques for Gold-granule Beads Excavated from the Neungsan-ri Temple Site in Buyeo (부여 능산리사지 출토 금제구슬의 재료학적 특성 및 제작기법 연구)

  • Yang, Soohyeon;Ro, Jihyun
    • Conservation Science in Museum
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    • v.26
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    • pp.67-82
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    • 2021
  • Two golden beads (Buyeo 5336) housed at the Buyeo National Museum were discovered in 1993 near the site of an ancient workshop in Neungsan-ri in Buyeo-gun, Chungcheongnam-do Province. These rare examples from the Baekje Kingdom of an application of granulation have maintained their original form intact, and thus serve as important materials for the investigation of production techniques applied. This study analyzed the composition of the golden beads using a portable X-ray fluorescence analyzer, a stereo microscope, and a scanning electron microscope with an energy dispersive X-ray spectrometer. The manufacturing technique was examined through the observation of the micro-shape and the surface condition and by a composition analysis of the joint part. In both beads, a hole was pierced in a hollow body and the bead was decorated with golden wires around the hole and gold granules in other parts. In some areas, golden granules had been attached to the gold plate and golden wires were then placed over the granules. The purity of both the wires and the granules was analyzed as 23.6 - 23.7K. A high copper content was detected in some of the parts where the granules were attached. The findings of a previous reproduction experiment and study of production methods suggest that the beads were made using the copper diffusion technique.

The study on cytotoxicity of cytokines produced by the activated human NKT cells on neuroblastoma (활성화된 자연살상 T 세포(NKT)에서 생성된 사이토카인에 의한 신경모세포종의 세포독성에 관한 연구)

  • Cho, Jin Young;Yoon, Young Wook;Yoon, Hyang Suk;Kim, Jong Duk;Choi, Du Young
    • Clinical and Experimental Pediatrics
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    • v.49 no.4
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    • pp.439-445
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    • 2006
  • Purpose : ${\alpha}$-Galactosylceramide (${\alpha}$-GalCer)-stimulated human $V{\alpha}24$ natural killer T (NKT) cells exert antitumor activity against some leukemia in a CD1d dependent and TCR-mediated manner, but could not kill CD1d - negative neuroblastoma (NB) cells. There are few reports about the direct antitumor effect of highly secreted cytokines by these cells on activation. In this study, using a cell-free supernatant (SPN) collected from plate bound hCD1d/${\alpha}$ GalCer tetramers-stimulated NKT cells, we examined whether they could be helpful in the immunotherapeutic treatment of NB. Methods : Cells were cultured in IMDM. The cytokines produced by NKT cells were measured with Cytometric Bead Array (CBA) analysis. Cell viability was evaluated by calcein-AM fluorescence with digital image microscopy scanning (DIMSCAN). The percentage of specific apoptosis was calculated by flow cytometric detection of apoptosis using annexin V and 7-AAD. Results : The activated NKT cells secreted high levels of IL-2, INF-${\gamma}$, TNF-${\alpha}$. The SPN was significantly cytotoxic against four out of eight tested NB cell lines, through mainly apoptosis as evidenced by annexin-V staining and inhibition with the pretreatment of pancaspase blocker. This apoptosis was significantly inhibited when anti-TNF-${\alpha}$ and anti-IFN-${\gamma}$ neutralizing mAbs were used separately and it was completely abolished when the two mAbs were combined. Conclusion : IFN-${\gamma}$ and TNF-${\alpha}$ produced by NKT cells could exert synergistically direct antitumor activity through apoptosis on some NB cell lines.

Carbon Monoxide Inhibits PMA-induced Differentiation in Human Monocytic THP-1 Cells (단핵구세포주 THP-1의 대식세포로의 분화 및 활성화에서 CO의 억제 효과)

  • Kim, Da Sol;Lee, Mi Sun;Kim, Han Sol;Lee, Hye Yun;Kim, Oh Yun;Kang, Ye Rin;Sohn, Dong Hyun;Kim, Koanhoi;Park, Young Chul
    • Journal of Life Science
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    • v.27 no.2
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    • pp.217-224
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    • 2017
  • Carbon monoxide (CO), a reaction product of cytoprotective enzyme heme oxygenase-1 (HO-1), is a gaseous messenger with anti-proliferative, anti-apoptotic, and anti-inflammatory actions in many cell types. Here, we investigated the role of CO on the process of monocyte differentiation induced by phorbol 12-myristate 13-acetate (PMA) in human monocytic THP-1 cells. CORM-2 (tricarbonyldichlororuthenium (II) dimer, $Ru2Cl_4(CO)_6$), a CO-releasing compound, decreased a marked cell adherence with a slight reduction of proliferation in monocytic THP-1 cells treated with PMA. And, CORM-2 significantly inhibited expression of differentiation markers such as CD14, CD11b plus CD18 (macrophage-1 antigen, Mac-1 or complement receptor 3, CR3) and phagocytosis of carboxylate-modified red fluorescent latex beads, in PMA-stimulated THP-1 cells. For the further experiments, differentiation of PMA-treated cells was enhanced after the initial 2 days stimulus by removing the PMA-containing media then incubating the cells in fresh media for a another 4 days. And, we observed the secretion of inflammatory cytokines and phagocytosis in differentiated macrophages. Treatment with CORM-2 significantly abolished the secretion of IL-6, $TNF-{\alpha}$ and phagocytosis using fluorescence-conjugated E. coli (K-12 strain) bioparticles in lipopolysaccharide (LPS)-stimulated differentiated macrophages. In conclusion, these results suggest that CO inhibits the differentiation of monocytic THP-1 cells as well as the activation of differentiated macrophages.

An Analysis of the Characteristics of Glass Beads from the Joseon Dynasty Using Non-destructive Analysis (비파괴 분석을 활용한 조선시대 유리구슬의 특성 분석)

  • Lee Sujin;Kim Gyuho
    • Conservation Science in Museum
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    • v.30
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    • pp.71-88
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    • 2023
  • This paper examined the visible characteristics and chemical composition of glass beads from the Joseon Dynasty as well as the associations thereof. It also explored the characteristics and uses of glass beads by region. This study covered a total of 1,819 pieces excavated from 25 locations in the Gyeonggi, Chungcheong, and Gyeongsang regions, of which 537 pieces were analyzed for their chemical composition. Glass beads of the Joseon Dynasty take a variety of shapes such as a Round, Coil, Floral, Segmented, Flat, Oval, and Calabash. Colors vary from shades of brown (brown, lemon yellow) and shades of blue (Bluish-Green, greenish-Blue, Purple-Blue) to shades of white (colorless, white) and shades of green (Green, Greenish-Blue, Greenish-Brown). Brown accounts for the largest percentage, followed by Bluish-Green, greenish-Blue. It was identified that Drawing technique was the most common glass bead production technique of the Joseon Dynasty. Potassium oxide (K2O) was the most common flux agent for glass beads, while the potash glass and mixed alkali glass groups account for the largest quantity. The choice of stabilizers depended on the type of flux agents used, but the most common were calcium oxide (CaO) and aluminum oxide (Al2O3). The potash glass and potash lead glass groups are high in CaO and low in Al2O3, the mixed alkali glass group is high in CaO, and the lead glass group is low in CaO. In terms of the association between color and shape, most of the beads with shade of brown and blue have round shapes of brown and blue have spherical shapes, while the coil shape is prominent in blue beads. A high percentage of green and colorless beads also take the shape of a coil, while white beads in general have a floral shape. In terms of the association between shape and chemical composition, round, floral and segmented shapes account for a high percentage of the potash glass group, while coil and flat shapes are common in the mixed alkali glass group. This paper also analyzed the colorants for each color based on the association between color and chemical composition. Iron (Fe) was used as the colorant for brown and white, and titanium (Ti) and iron were used for light yellow. Purple-Blue was produced by by cobalt (Co), and greenish-Blue, Bluish-Green, green, Greenish-Blue were produced by iron and copper (Cu). Colorless beads had a generally low colorant content.