• Title/Summary/Keyword: Blood Assay

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Immuno-enhancing Effect of Protaetia brevitarsis seulensis (white grub) Extracts on RAW 264.7 Cell Line. (굼벵이 추출물의 RAW 264.7 세포에 대한 면역증강 효과)

  • Park, Bog-Im;Seo, Jae-Bin;Jin, Yu-Mi;Kim, Seong-Seon;Sim, Hyeon-Jae;Lee, Hoon-Yeon;Kim, Seong-Oh;kim, Dong-Keun;Jo, Mi-Na;Cho, Yong-Jin;Kim, Chong-Tai;Kim, Tae-Eun;Jeon, Yong-Deok;Jin, Jong-Sik
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.102-102
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    • 2018
  • Protaetia Brevitarsis Seulensis(white grub) has been traditionally used as medicinal stuff to treat blood stasis, occlusion of menstruation, tetanus and liver cancer in Asian countries (Korea, Japan, China, Taiwan, India and Myanmar). Especially, Donguibogam, which is traditional korean medicinal book, described the white grub as traditional medicine to treat hepatic diseases and vascular disorders. The white grub has been considered as highly nutritional food. The major constituents of white grub are rich in protein, healthy fats, iron, calcium. Recent studies announced that white grub has hepatoprotective effect and anti-microbacterial effect. However, the immuno-enhancing effect of white grub extracts in RAW 264.7 macrophage cells has not been studied yet. In this study, the various concentrations of white grub extract were examined to find immuno-enhancing effects on RAW 264.7 cells. Cytotoxicity was determined by MTT assay and immuno-enhancing effect of white grub extract was investigated by measuring nitric oxide (NO) production compared with only lipopolysaccharide (LPS) treatment. White grub extracts (0.001 - 10 mg/ml) did not show cytotoxicity. Additionally, white grub extracts (0.001 - 1mg/ml) had Immuno-enhancing effect on RAW 264.7 cells compared with only LPS treated group. These results might be provided proof to develop beneficial immuno-enhancing material for human health.

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Induction of Micronuclei in Human and Mouse Lymphocytes Irradiated with Gamma Radiation and Effect of Panax ginseng C.A. Meyer (마우스와 사람 림프구에서 방사선에 의한 미소핵의 형성 및 고려인삼의 효과)

  • Kim, Sung-Ho;Oh, Heon;Lee, Song-Eun;Lee, Yun-Sil;Kim, Tae-Hwan;Jeong, Kyu-Sik;Ryu, Si-Yun
    • Journal of Radiation Protection and Research
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    • v.22 no.3
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    • pp.153-160
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    • 1997
  • The frequencies of ${\gamma}$-ray-induced micronuclei (MN) in cytokinesis-blocked (CB) lymphocytes at several doses were measured in three donors of human and C57BL/6 mice. Measurements performed after irradiation showed a dose-related increases in MN frequency in each of the donors studied. The relative sensitivity of mouse in spleen lymphocytes (SLs) compared with human peripheral blood lymphocytes (PBLs) was estimated by best fitting linear-quadratic model based on the radiation-induced MN data over the range from 0 cGy to 400 cGy. In the case of MN frequency with 0.2 per CB cell, the relative sensitivity of mouse SLs was 1.67. Compared with the radiation-induced MN formation in the PBLs of human, the SLs of mouse were more radiosensitive. Using this MN assay with human PBLs and mouse SLs, studies were performed to determine whether the water fraction of ginseng (Panax ginseng C.A.Meyer) against radiation-induced MN in human PBLs after in vitro irradiation (3Gy) and in SLs of C57BL/6 mice after in vivo irradiation (3Gy). The frequency of MN in human PBLs was reduced by water fraction of ginseng (0.5mg/ml of medium) both pre-and post treatment (p<0.0l) in vitro. In addition, the frequency of MN in mouse SLs was also reduced by pretreatment of ginseng (2mg/ml of drinking water for 7days) in vivo. The data suggested that the ginseng may reduce cell damage caused by ${\gamma}$-rays in vitro and in vivo. Further studies are needed to characterize better the protective nature of ginseng extract, its fractions and compounds.

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Acridine Orange Stained Micronucleus Assay in Human B and T-lymphocytes after Low Dose ${\gamma}-irradiation$ (아크리딘 오렌지 형광염색법을 이용한 저선량 감마선 유도 말초혈액 B와 T-림프구 미소핵 분석)

  • Choi, Jeong-Mi;Kim, Hee-Sun;Yang, Kwang-Hee;Kim, Cha-Soon;Lim, Yong-Khi;Kim, Chong-Soon;Woon, Jae-Ho
    • Journal of Radiation Protection and Research
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    • v.29 no.1
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    • pp.9-15
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    • 2004
  • Firstly, we compared the two staining techniques, Giemsa and Acridine orange, to determine micronuclei on samples of cultures of five healthy human peripheral blood lymphocytes after ${\gamma}-irradiation\;(^{137}Cs)$ in dose ranges of 0 to 800cGy. It was found that the Acridine orange staining method gives more reliable results than the usual Giemsa staining method in micronucleus tests. Moreover, the frequency of micronuclei in cytokinesis-blocked human B-lymphocytes was studied after in vitro irradiation in dose ranges of 0 to 50cGy. After setting and separating the B-lymphocytes, the frequency of radiation-induced micronuclei were observed as the end-point markers for the low-dose radiation dosimetry after staining with Giemsa and Acridine orange dyes. The micronuclei frequency in B-lymphocytes was significantly elevated from 10 to 30cGy ${\gamma}-irradiation$. The determination of micronuclei in B-lymphocytes after staining with Acridine orange was higher than that of Giemsa. The frequency of micronuclei in B-lymphocytes was observed to be at least two times higher than those of T-lymphocytes Giemsa in dose increasing. Therefore, the determination of low-dose radiation-induced micronuclei in B-lymphocytes after staining with Acridine orange is likely to have the greatest potential in the estimation of low dose radiation exposure.

The Significance of Caspase-Cleaved Cytokeratin 18 in Pleural Effusion

  • Lee, Keu Sung;Chung, Joo Yang;Jung, Yun Jung;Chung, Wou Young;Park, Joo Hun;Sheen, Seung Soo;Lee, Kyi Beom;Park, Kwang Joo
    • Tuberculosis and Respiratory Diseases
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    • v.76 no.1
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    • pp.15-22
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    • 2014
  • Background: Apoptosis plays a role in the development of pleural effusion. Caspase-cleaved cytokeratin 18, a marker for epithelial cell apoptosis, was evaluated in pleural effusion. Methods: A total of 79 patients with pleural effusion were enrolled. The underlying causes were lung cancer (n=24), parapneumonic effusion (n=15), tuberculous effusion (n=28), and transudates (n=12). The levels of M30, an epitope of caspase-cleaved cytokeratin 18, were measured in blood and pleural fluids using enzyme-linked immunosorbent assay along with routine cellular and biochemical parameters. The expression of M30 was evaluated in the pleural tissues using immunohistochemistry for M30. Results: The M30 levels in pleural fluid were significantly higher in patients with tuberculosis ($2,632.1{\pm}1,467.3U/mL$) than in patients with lung cancer ($956.5{\pm}618.5U/mL$), parapneumonic effusion ($689.9{\pm}413.6U/mL$), and transudates ($273.6{\pm}144.5U/mL$; all p<0.01). The serum levels were not significantly different among the disease groups. Based on receiver operating characteristics analysis, the area under the curve of M30 for differentiating tuberculous pleural effusion from all other effusions was 0.93. In the immunohistochemical analysis of M30, all pathologic types of cancer cells showed moderate to high expression, and the epithelioid cells in granulomas showed high expression in tuberculous pleural tissues. Conclusion: Caspase-cleaved cytokeratin 18 was most prominently observed in tuberculous pleural effusion and showed utility as a clinical marker. The main source of M30 was found to be the epithelioid cells of granulomas in tuberculous pleural tissues.

Study on the Influenza Type A Activity of Fractions of Korean Medicinal Herbs (한약 단미제 분획의 Influenza virus type A 활성에 관한연구)

  • Jung, Jae-Deuk;Park, In-Ho;Lee, Kwang-Hee;Kim, Ho-Kyoung;Ko, Byung-Seob;Park, Kap-Joo
    • Korean Journal of Oriental Medicine
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    • v.3 no.1
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    • pp.67-83
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    • 1997
  • In order to search for anti-influenza virus type A agents from Korean medicinal herbs, we selected 100 medicinal herbs, based on a review of the Korean traditional medicine books. Four of 100 Korean medicinal herbs, MM-40, MM-55, MM-63, MM-110, exhibited very strong anti-influenza virus activity. The fractions of four medicinal herbs, which had very strong anti-influenza virus activity, were tested for antiviral activity by means of Haemagglutination inhibition test(HTT), 40% MeOH fraction of MM-40, $H_2O$ fraction of MM-55, 20% fraction of MM-63 3nd $H_2O$ fraction of MM-110 had strong anti-influenza virus activity at the range of $78{\mu}g/ml$ to $156{\mu}g/ml$, 1.56mg/ml to 100mg/ml, 6.25mg/ml to 50mg/ml and $48.7{\mu}g/ml$ to $780{\mu}g/ml$, respectively. These results of HIT indicated that fractions of Korean medicinal herbs might inhibit either attachment of virus to cell surface receptor or penetration of virus into cell during the initial stage of infection. In the cytotoxicity of fractions against red blood cells, 40% MeOH fraction of MM-40, 20% fraction of MM-63 and $H_2O$ fraction of MM-110 showed cytotoxicity at the range of $78{\mu}g/ml$ to 10mg/ml, 50mg/ml to 100mg/ml and $195{\mu}g/ml$ to 100mg/ml, respectively, whereas $H_2O$ fraction of MM-55 did not show cytotoxicity. In order to establish influenza virus adapted animal model, influenza virus type A were passaged 3 and 4 times successively in Balb/c mouse. As a result, we had 4 HA unit titers on the 5 days of 3rd passages and 7 days of 4th passages after infection, respectively.

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BIOLOGICAL EFFECT OF MAGNOLIA AND GINKGO BILOBA EXTRACT TO THE ANTIMICROBIAL, ANTIINFLAMMATORY AND CELLULAR ACTIVITY (후박 및 은행잎 추출물의 향균, 향염 및 세포활성도에 미치는 영향)

  • Chung, Chong-Pyuong;Ku, Young;Bae, Ki-Hwan
    • Journal of Periodontal and Implant Science
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    • v.25 no.3
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    • pp.478-486
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    • 1995
  • Periodontal therapy for treatment of periodontitis involves the elimination of bacterial plaque and elimination of the anatomic defects by regenerative procedure. The purpose of this study was to evaluate on the biological effect of magnolia and Ginkgo biloba extract to the antimicrobial, antiinflammatory and cellular activity. Antimicrobial assay was performed with the diffusion method of the extract by measuring of growth inhibitory zone of B. cereus from blood agar plate. Effect of the extract to cellular activity of gingival fibroblast were examined using MTT method and measured the result with optical density on 570nm by ELISA reader. Inhibitory effects of $PGE_2$ production from gingival fibroblast was performed with the addition of $IL-l{\beta}$ and the extract to the well and examined to the product of $PGE_2$ from cell by ELISA reader. In vivo anti-inflammatory effect was performed with injection examined with clinically and histologically for their extent of mecrosis and inflammation. Antimicrobial activity of Magnolia extract showed significantly higher activity than that of control. However, GBE did not showed significant activity to compare with control, and mixture of Magnolia and GBE extract showed significantly higher activity than that of control. The effect of cellular activity to gingival fibroblast showed no significant differences of between control and Magnolia extract. However, GBE showed significantly higher rate of cellular activity to compare with control and even to PDGF-BB, and also showed same degree of cellular activity even though mixed with Magnolia extract. The inhibitory effect of $PGE_2$ production showed significantly reduction of $PGE_2$ production to compare with control, but its inhibitory effect was not much strong to compare with Indomethacin. In vivo, antiinflammatory effect of Magnolia extract to P. gingivalis injection of Hamster buccal check showed significantly reduction of inflammatory cell infiltration and tissue necrosis, but GBE showed no effect on the inhibition of inflammatory process. These results suggested that Magnolia and GBE extract possessed different kind of biological activity and also can be compensated on their activity with each other for elimination of bacterial plaque and anatonical defect.

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Regulatory effects of Seogakjihwang-tang on Cytokines and Growth Factor Production in PBMC from the Patient with Cerebral infarction under Consciousness Disorders

  • Kim Yo Han;Sung Kang Keyng;Lee Kwang Ro;Lee Sang Kwan;Cheong Sang Su;Kang Sei Young;Lee So Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.3
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    • pp.829-836
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    • 2003
  • Seogakjihwang-tang (SJT) was widely used to treat patients suffering from cerebral infarction. But scientific investigation has been carried out very little. The aim of the present study is to investigate the effect of SJT on the production of various cytokines in the patients with cerebral infarction (CI). We investigated interleukin (IL)-4, IL-10 and transforming growth factor (TGF)-1 in the sera of 27 patients with cerebral infarction under consciousness disorders and 10 normal controls using an originally devised sensitive sandwich enzyme-linked immunosorbent assay (ELISA). We found that plasma levels of IL-4 were slightly elevated in patients with cerebral infarction, whereas plasma levels of IL-10 (P<0.001) and TGF-1 were reduced. Peripheral blood mononuclear cells (PBMC) obtained from the patient with CI were cultured for 24 h in the presence or absence of lipopolysaccharide (LPS) or phytohaemagglutinin (PHA). The amount of IL-4, IL-10 and TGF-1, in culture supernatant, was significantly increased in the LPS or PHA treated cells compared to unstimulated cells (P<0.05), We also show that increased cytokines IL-4, and IL-10 level was significantly inhibited by SJT in a dose-dependent manner. Maximal inhibition rate of IL-4 and IL-10 production by SJT was 45.63.3% and 614.7% for LPS-stimulated cell and 27.31.2% and 83.62% for PHA-stimulated cells, respectively (P<0.05). On the other hand, SJT significantly increased the LPS or PHA-induced TGF-1 production (P<0.05). These data suggest that SJT has a regulatory effect on the cytokines production, which might explain its beneficial effect in the treatment of CI.

Anti-diabetic Effects of Banaba Leaf Extracts (Lagerstroemia speciosa Pers.) through Solvents (추출용매 변화에 따른 바나바(Lagerstroemia speciosa Pers.) 잎 추출물의 항당뇨 효과)

  • Kim, Kyun-Ha;Roh, Sang-Geun;Li, Chun-Ri;Jin, Chun-Feng;Kim, Andre;Choi, Won-Chul
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1305-1311
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    • 2008
  • This study investigated that the antidiabetic effects of banaba extracts with variety solvents selectivity in vitro and in vivo. Banaba extracts were prepared with water, 70% ethanol, 90% ethanol, 100% ethanol and water-ethanol that of extract twice times sequentially water and ethanol. Cell toxicity and insulin secretion of banaba extracts was tested by MTT (3-[4,5-dimetylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay on hamster insulinoma cell line, HIT-T15. Also we tested that insulin, body weight, blood glucose, total cholesterol, HDL-cholesterol, triglyceride and free fatty acid in streptozotocin diabetic rats. Water-ethanol extract has remarkable antidiabetic effect compare with the other banaba extracts. For water-ethanol extract has both of hydrophilic and hydrophobic antidiabetic materials from banaba. Expecially, corosolic acid, as known as unique polyphenol, has antidiabetic effect studied by many researchers till nowadays. But corosolic acid does not solve in water. Otherwise, we suggest that banaba extract of hydrophilic and hydrophobic materials (polyphenol and antioxidants) mixture more increased antidiabetic effects.

The Significance of SDF-1α-CXCR4 Axis in in vivo Angiogenic Ability of Human Periodontal Ligament Stem Cells

  • Bae, Yoon-Kyung;Kim, Gee-Hye;Lee, Jae Cheoun;Seo, Byoung-Moo;Joo, Kyeung-Min;Lee, Gene;Nam, Hyun
    • Molecules and Cells
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    • v.40 no.6
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    • pp.386-392
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    • 2017
  • Periodontal ligament stem cells (PDLSCs) are multipotent stem cells derived from periodontium and have mesenchymal stem cell (MSC)-like characteristics. Recently, the perivascular region was recognized as the developmental origin of MSCs, which suggests the in vivo angiogenic potential of PDLSCs. In this study, we investigated whether PDLSCs could be a potential source of perivascular cells, which could contribute to in vivo angiogenesis. PDLSCs exhibited typical MSC-like characteristics such as the expression pattern of surface markers (CD29, CD44, CD73, and CD105) and differentiation potentials (osteogenic and adipogenic differentiation). Moreover, PDLSCs expressed perivascular cell markers such as NG2, ${\alpha}-smooth$ muscle actin, platelet-derived growth factor receptor ${\beta}$, and CD146. We conducted an in vivo Matrigel plug assay to confirm the in vivo angiogenic potential of PDLSCs. We could not observe significant vessel-like structures with PDLSCs alone or human umbilical vein endothelial cells (HUVECs) alone at day 7 after injection. However, when PDLSCs and HUVECs were co-injected, there were vessel-like structures containing red blood cells in the lumens, which suggested that anastomosis occurred between newly formed vessels and host circulatory system. To block the $SDF-1{\alpha}$ and CXCR4 axis between PDLSCs and HUVECs, AMD3100, a CXCR4 antagonist, was added into the Matrigel plug. After day 3 and day 7 after injection, there were no significant vessel-like structures. In conclusion, we demonstrated the perivascular characteristics of PDLSCs and their contribution to in vivo angiogenesis, which might imply potential application of PDLSCs into the neovascularization of tissue engineering and vascular diseases.

Effects of Helicobacter pylori Antigen on Producton of Transforming growth factor-$\beta$1 and Nitric oxide in Human Fibroblast (사람성유아세포의 Transforming growth factor-$\beta$1과 Nitric oxide 생성에 미치는 Helicobacter pylori 항원의 효과)

  • 박무인;박선자;구자영;김광혁
    • Journal of Life Science
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    • v.11 no.2
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    • pp.181-189
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    • 2001
  • Cytokines are hormone-like proteins which mediate and regulast inflammatory and immune responses. Transforming growth factor -$\beta$1(TGF-$\beta$) plays an important role in the control of the immune response and wound healing, and in the development o various tissues and organs, Nitric oxide(NO) is major messenger molecule regulating immune function and blood vessel dilation and serving as a neurotransmitter in the brain and peripheral nervous system. Also, NO is to be a potent mutagen that cause mutation in the p53 tumor suppressor gene in early phases of human gastric carcinogenesis. The purpose of this study was to investigate the effect of Helicobacter phlori lystes, lipopolysaccharide (LPS), and Staphylococcus enterotoxin B(SEB) on production of TGF-$\beta$1 and NO by human fibroblasts. Primary cultured human fibroblasts were incubated with H. pylori lysates(Hp), LPs, SEB, Hp+LPS, Hp+SEB, Hp+LPS+SEB. Cultured supernatants that were collected at 24, 48 and 72 hr were assessed for TGF-$\beta$1 by enzyme-linked immunosorbent assay and NO production by quantification of nitrite ion. TGF-$\beta$1 production in fibroblasts exposed with Hp, LPS or SEB for 48 hrs was enhanced, but for 72 hrs inhibited. Its production by doble exposure such as Hp+LPS, Hp+SEB, Hp+LPS+SEB was lowered in comparison with single exposure of Hp in cases of 24 and 48 hrs incubation, but for 72 hrs decreased in Hp vaculoating toxin(+), increased in Hp vacuolating toxin(-). No production in fibroblasts increaed at all doses of LPS. But its production by exposure of SEB increased or decreased according to dose and incubation time. Also, NO production by Hp vacuolating toxin(+) increased at all doses, but its production by Hp vacuolating toxin(-) decreased. Its production by doble exposure such as Hp+LPS, Hp+SEB, Hp+LPS+SEB decreased in comparison with single exposure Hp Therefore, quantities pf TGB-$\beta$1 and NO released by human fibroblasts shows differences according to kinds of stimulants. Also, in care stimulated with same kinds of stimulants, its productions exhibit quantitative differences according to exposure times. These results suggest that the decreased of TGF-$\beta$1 in fibroblasts by mixed exposure with Hp producing vacuolating toxin and bacterial toxins such as LPS and SEB may effect negatively in healing of host tissue and increased of NO by infection oh H. pylori may related to the increased susceptibility for human gastric carcinogenesis.

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