• Title/Summary/Keyword: Cell assay

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Clinical Application of Serum CEA, SCC, Cyfra21-1, and TPA in Lung Cancer (폐암환자에서 혈청 CEA, SCC, Cyfra21-1, TPA-M 측정의 의의)

  • Lee, Jun-Ho;Kim, Kyung-Chan;Lee, Sang-Jun;Lee, Jong-Kook;Jo, Sung-Jae;Kwon, Kun-Young;Han, Sung-Beom;Jeon, Young-June
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.785-795
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    • 1997
  • Background : Tumor markers have been used in diagnosis, predicting the extent of disease, monitoring recurrence after therapy and prediction of prognosis. But the utility of markers in lung cancer has been limited by low sensitivity and specificity. TPA-M is recently developed marker using combined monoclonal antibody of Cytokeratin 8, 18, and 19. This study was conducted to evaluate the efficacy of new tumor marker, TPA-M by comparing the estabilished markers SCC, CEA, Cyfra21-1 in lung cancer. Method : An immunoradiometric assay of serum CEA, sec, Cyfra21-1, and TPA-M was performed in 49 pathologically confirmed lung cancer patients who visited Keimyung University Hospital from April 1996 to August 1996, and 29 benign lung diseases. Commercially available kits, Ab bead CEA (Eiken) to CEA, SCC RIA BEAD (DAINABOT) to SCC, CA2H (TFB) to Cyfra2H. and TPA-M (DAIICHI) to TPA-M were used for this study. Results : The mean serum values of lung cancer group and control group were $10.05{\pm}38.39{\mu}/L$, $1.59{\pm}0.94{\mu}/L$ in CEA, $3.04{\pm}5.79{\mu}/L$, $1.58{\pm}2.85{\mu}/L$ in SCC, $8.27{\pm}11.96{\mu}/L$, $1.77{\pm}2.72{\mu}/L$ in Cyfra21-1, and $132.02{\pm}209.35\;U/L$, $45.86{\pm}75.86\;U/L$ in TPA-M respectively. Serum values of Cyfra21-1 and TPA-M in lung cancer group were higher than control group (p<0.05). Using cutoff value recommended by the manufactures, that is $2.5{\mu}/L$ in CEA, $3.0{\mu}/L$ in Cyfra21-1, 70.0 U/L in TPA-M, and $2.0{\mu}/L$ in SCC, sensitivity and specificity of lung cancer were 33.3%, 78.6% in CEA, 50.0%, 89.7% in Cyfra21-1, 52.3%, 89.7% in TPA-M, 23.8%, 89.3% in SCC. Sensitivity and specificity of nonsmall cell lung cancer were 36.1%, 78.1% in CEA, 50.1%, 89.7% in Cyfra21-1, 53.1%, 89.7% in TPA-M, 33.8%, 89.3% in SCC. Sensitivity and specificity of small cell lung cancer were 25.0%, 78.5% in CEA, 50.0%, 89.6% in Cyfra21-1, 50.0%, 89.6% in TPA-M, 0%, 89.2% in SCC. Cutoff value according to ROC(Receiver operating characteristics) curve was $1.25{\mu}/L$ in CEA, $1.5{\mu}/L$ in Cyfra2-1, 35 U/L in TPA-M, $0.6{\mu}/L$ in SCC. With this cutoff value, sensitivity, specificity, accuracy and kappa index of Cyfra21-1 and TPA-M were better than CEA and SCC. SCC only was related with statistic significance to TNM stages, dividing to operable stages(TNM stage I to IIIA) and inoperable stages (IIIB and IV) (p<0.05). But no tumor markers showed any correlation with significance with tumor size(p>0.05). Conclusion : Serum TPA-M and Cyfra21-1 shows higher sensitivity and specificity than CEA and SCC in overall lung cancer and nonsmall cell lung cancer those were confirmed pathologically. SCC has higher specificity in nonsmall cell lung cancer. And the level of serum sec are signiticantly related with TNM staging.

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Plasma G-CSF and GM-CSF Concentrations and Expression of their Receptors on the Granulocyte in Children with Leukocytosis (백혈구 증가증 환아의 혈장내 G-CSF와 GM-CSF의 농도 및 과립구에서의 이들 수용체의 발현)

  • Choi, Won Seok;Ryu, Kyung Hwan;Kim, You Jeong;Kim, So Young;Kim, Hyun Hee;Lee, Wonbae
    • Clinical and Experimental Pediatrics
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    • v.46 no.3
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    • pp.271-276
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    • 2003
  • Purpose : Granulocyte-colony stimulating factor(G-CSF) and granulocyte macrophage-colony stimulating factor(GM-CSF) are principal cytokines in granulopoiesis and their physiologic effects are mediated through binding to specific cell surface receptors. Although it is known that the level of serum G-CSF and GM-CSF, and presentation of the receptors are increased in infectious diseases, there have been no studies to find the correlation between the granulopoiesis and leukocytosis. This study was designed to measure G-CSF and GM-CSF in leukocytosis and in control and to demonstrate the possible pathogenesis of granulopoiesis in leukocytosis using quantitative analysis of G-CSF, GM-CSF and their CSFr. Methods : The plasma levels of G-CSF, GM-CSF of 13 children without leukocytosis and 14 children with leukocytosis were measured. Counts of cell surface G-CSFr and GM-CSFr were measured by combining anti G-CSFr and anti GM-CSFr monoclonal antibodies to their respective receptors by using quantitative flow cytometric assay. Results : There was no significant difference betweeen the plasma concentration of G-CSF and GM-CSF in acute leukocytosis and in the control group. However, levels of G-CSFr in acute leukocytosis decreased significantly compared to the control(P=0.012) and the levels of GM-CSFr in both groups revealed no significant difference. Conclusion : Increase in the number of leukocyte in leukocytosis was mediated by increasing the number of neutrophil, and increased plasma concentration of G-CSF may be the cause of neutrophilia. But GM-CSF did not have any influence on leukocytosis.

Effect of Boswellia serrata Extracts on Degenerative Osteoarthritis in vitro and in vivo Models (보스웰리아 추출물의 골관절염 억제 효과 연구)

  • Nam, Da-Eun;Kim, Ok Kyung;Shim, Tae Jin;Kim, Ji Hoon;Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.5
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    • pp.631-640
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    • 2014
  • The inhibitory effects of Boswellia serrata (BW) extracts on degenerative osteoarthritis were investigated in primary-cultured rat cartilage cells and a monosodium-iodoacetate (MIA)-induced osteoarthritis rat model. To identify the protective effects of BW extract against $H_2O_2$ ($800{\mu}M$, 2 hr) in vitro, cell survival was measured by MTT assay. Cell survival after $H_2O_2$ treatment was elevated by BW extract at a concentration of $20{\mu}g/mL$. In addition, BW extract treatment significantly reduced and normalized the productions of pro-inflammatory factors, nuclear transcription factor ${\kappa}B$, cyclooxygenase-2, tumor necrosis factor-${\alpha}$, and interleukin-6 at a concentration of $20{\mu}g/mL$. Treatment of chondrocytes with BW extract significantly reduced 5-lipoxygenase activity and production of prostaglandin E2, especially at a concentration of $10{\sim}20{\mu}g/mL$. For the in vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats. Consumption of a diet containing BW extract (100 and 200 mg/kg) for 35 days significantly inhibited the development and severity of osteoarthritis in rats. To determine the genetic expression of arthritic factors in articular cartilage, real-time PCR was applied to measure matrix metalloproteinases (MMP-3, MMP-9, and MMP-13), collagen type I, collagen type II, and aggrecan, and BW extract had protective effects at a concentration of 200 mg/kg. In conclusion, BW extract was able to inhibit articular cartilage degeneration by preventing extracellular matrix degradation and chondrocyte injury. One can consider that BW extract may be a potential therapeutic treatment for degenerative osteoarthritis.

The Signal Transduction Mechanisms on the Intestinal Mucosa of Rat Following Irradiation (방사선조사후 백서소장점막에서 발생하는 신호전달체계에 관한 연구)

  • Yoo Jeong Hyun;Kim Sung Sook;Lee Kyung Ja;Rhee Chung Sik
    • Radiation Oncology Journal
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    • v.15 no.2
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    • pp.79-95
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    • 1997
  • Purpose : Phospholipase C(PLC) isozymes play significant roles in signal transduction mechanism. $PLC-\gamma$ 1 is one of the key regulatory enzymes in signal transduction for cellular proliferation and differentiation. Ras oncoprotein, EGFR, and PKC are also known to be involved in cell growth. The exact mechanisms of these signal transduction following irradiation, however, were not clearly documented Thus, this study was Planned to determine the biological significance of PLC, ras oncoprotein, EGFR, and PKC in damage and regeneration of rat intestinal mucosa following irradiation. Material and Method : Sixty Sprague-Dawley rats were irradiated to entire body with a single dose of 8Gy. The rats were divided into S groups according to the sacrifice days after irradiation. The expression of PLC, ras oncoprotein, EGFR and PKC in each group were examined by the immunoblotting and immunohistochemistry. The histopathologic findings were observed using H&I stain, and the mitoses for the evidence of regeneration were counted using the light microscopy & PCNA kit. The Phosphoinositide(PI) hydrolyzing activity assay was also done for the indirect evaluation of $PLC-\gamma$ 1 activity. Results: In the immunohistochemistry , the expression of $PLC-{\beta}$ was negative for all grøups. The expression of $PLC-{\gamma}1$ was highest in the group III followed by group II in the proliferative zone of mucosa. The expression of $PKC-{\delta}1$ was strongly positive in group 1 followed by group II in the damaged surface epithelium. The above findings were also confirttled in the immunoblotting study. In the immunoblotting study, the expressions of $PLC-{\beta}$, $PLC-{\gamma}1$, and $PKC-{\delta}1$ were the same as the results of immunohis-tochemistry. The expression of ras oncoprctein was weakly positive in groups II, III and IV. The of EGFR was the highest in the group II, III, follwed by group IV and the expression of PKC was weakly positive in the group II and III. Conclusion: $PLC-{\gamma}1$ mediated signal transduction including ras oncoprotein, EGFR, and PKC play a significant role in mucosal regeneration after irradiation. $PLC-{\delta}1$ mediated signal transduction might have an important role in mucosal damage after irradiation. Further studies will be necessary to confirm the signal transduction mediating the $PKC-{\delta}1$.

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Effect of Solvent Fractions from Doenjang on Antimutagenicity, Growth of Tumor Cells and Production of Interleukin-2 (된장 분획물의 항돌연변이 및 암세포 증식 억제효과와 interleukin-2 생성에 미치는 영향)

  • Kim, Kwang-Hyuk;Park, Kun-Young;Lee, Sook-Hee;Lim, Sun-Young
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.791-797
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    • 2007
  • We studied the inhibitory effect of solvent fractions from doenjang on mutagenicity using Salmonella typhimurium TA100 in Ames test. We also investigated the effect of solvent fractions from doenjang on the growth of tumor cells and the production of interleukin-2 (IL-2). The treatment of dichlorormethane and ethylacetate fractions (2.5 mg/assay) from doenjang to Ames test system inhibited aflatoxin B$_1$ (AFB$_1$) induced mutagenicity by 96% and 97%, respectively, and showed a higher antimutagenic effect than other solvent fractions. In case of N-methyl-N'-nitro-N-nitrosoguamidine (MNNG) induced mutagenicity, the ethylacetate fraction showed the highest inhibitory effect (by 75%) among the other sol-vent fractions, although the inhibitory effect was not stronger compared to AFB$_1$ induced mutagenicity. The treatment of dichloromethane and ethylacetate fractions markedly inhibited the growth of Yac-1 (by 80% and 94%, respectively) and sacroma-180 cancer cells (by 60% and 96%, respectively) after 4 days of incubation at 37${\circ}$C. To elucidate the immunological mechanism of antitumor activity of doenjang, spleen cells of Balb/c mouse were exposed to the dichloromethane and ethyl-acetate fractions for 24 hours at 37${\circ}$C . The culture supernatants following the treatment of djchloromethane and ethylacetate factions to spleen cells increased the production of IL-2. These results indicated that the anticarcinogenic effect of doenjang was mediated by the production of IL-2.

The Study of Anti-inflammatory Effect of Suryeon-hwan Water Extract in RAW 264.7 Cells (대식세포에서 수련환(茱連丸) 물추출물의 항염증작용에 관한 연구)

  • Yoon, Yeo-Hwan;Kim, Sung-Bae;Kang, Ok-Hwa;Mun, Su-Hyun;Seo, Yun-Soo;Yang, Da-Wun;Kang, Da-Hye;Wi, Gyeong;Lim, Jae-Soo;Kim, Ma-Ryong;Kwak, Nam-Won;Kong, Ryong;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.29 no.6
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    • pp.125-132
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    • 2014
  • Objectives : Suryeon-hwan (SRH) exhibits potent anti-inflammatory activity with an unknown mechanism. However, there has been a lack of studies regarding the effects of SRH on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. So, the investigation focused on whether SRH inhibited nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) productions, as well as the expressions of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with 200 ng/mL of LPS 30 min prior to the addition of SRH. Cell viability was measured by MTS assay. The production of nitric oxide (NO) was determined by reacting cultured medium with Griess reagent. The content of level of cytokines (PGE, IL-6) in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. Results : We found that SRH inhibited LPS-induced NO, $PGE_2$ and IL-6 productions as well as the expressions of iNOS and COX-2. Furthermore, SRH suppressed the LPS-induced phosphorylation of MAPK and extracellular signal-regulated kinase 1/2 (ERK 1/2) activation. Conclusions : These results suggest that SRH has inhibitory effects on LPS-induced $PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the phosphorylation of MAPKs following activation.

Antioxidative Effect of Extracts from Different Parts of Juncus effusus L. (골풀 부위별 추출물의 항산화 효과)

  • Choi, Chang-Hwan;Won, Doo-Hyun;Hwang, Jun-Pil;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.3
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    • pp.275-282
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    • 2012
  • In this study, the antioxidative effects of extracts from different parts of Juncus effusus L. were investigated. The three parts (above-ground part, below-ground part, medulla part) were selected. 50 % ethanol extract, ethyl acetate and aglycone fractions of J. effusus L. were used in experiments. The highest DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activities ($FSC_{50}$) was shown by medulla part (42.9 ${\mu}g/mL$) in 50 % ethanol extracts, below-ground part (12.1 ${\mu}g/mL$) in ethyl acetate fractions, and below-ground part (12.1 ${\mu}g/mL$) in aglycone fractions. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system using the luminol-dependent chemiluminescence assay showed the most prominent effect of medulla part (0.29 ${\mu}g/mL$) in 50 % ethanol extracts, below-ground part (0.25 ${\mu}g/mL$) in ethyl acetate fractions, and medulla part (0.20 ${\mu}g/mL$) in aglycone fractions. The cellular protective effects of extract/fractions of J. effusus L. on the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner (0.5 ~ 10 ${\mu}g/mL$). Especially, aglycone fraction of medulla part at a concentration of 10 ${\mu}g/mL$ showed the most prominent protective effect among all extracts (${\tau}_{50}$, 321.0 min). These results indicate that extracts from below-ground part and medulla part of J. effusus L. extracts can be used as an natural antioxidant. Particularly, J. effusus L. can protect suggesting a high ${\tau}_{50}$ skin where many $^1O_2$ was generated by sunlight exposure.

Biological Control of Garlic Blue Mold using Pantoea agglomerans S59-4 (Pantoea agglomerans S59-4를 이용한 마늘 푸른곰팡이병의 생물학적 방제)

  • Kim, Yong-Ki;Hong, Sung-Jun;Jee, Hyung-Jin;Park, Jong-Ho;Han, Eun-Jung;Park, Kyung-Seok;Lee, Sang-Yeob;Lee, Seong-Don
    • The Korean Journal of Pesticide Science
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    • v.14 no.2
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    • pp.148-156
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    • 2010
  • S59-4 isolate was evaluated as a potential biocontrol agent using in vivo wounded garlic bulb assay. When the spore suspension ($10^5$ spores/$m\ell$) of Penicillium hirsutum was co-inoculated with cell suspension of S59-4 isolate on wounded garlics, the isolate showed high suppressive effect to disease development. The isolate was identified as Pantoea agglomerans S59-4(Pa59-4) through Biolog system. Furthermore, soaking garlic bulbs in the suspension of Pa59-4 significantly reduced garlic decay caused by P. hirsutum. The optimal concentration of Pa59-4 for controlling garlic blue mold was $10^7\sim10^8$ cfu/$m\ell$. And suppressive effect of Pa59-4 on garlic storage decay reduced as inoculation concentration of Penicillium hirsutum increased. In addition in order to investigate population dynamics of Pa59-4 on application site of garlic cloves, two antibiotic markers, pimaricin and vancomycin were selected. Bacterial density of Pa59-4 on the wounded garlic cloves increased continuously both under room temperature condition and low temperature condition until 30days after application of Pa59-4, meanwhile that of Pa59-4 on intact garlic cloves increased until 15days after application of Pa59-4 and thereafter decreased continuously. Two culture media for mass-production of Pa59-4, LB medium and TSB medium, were selected. By-product of bio-fungicide formulated by mixing white carbon and bacterial suspension of Pa59-4 suppressed by 40 to 50% garlic blue mold. Above results suggest that Pa59-4 be a promising control agent against garlic blue mold.

Stem-leaf saponins from Panax notoginseng counteract aberrant autophagy and apoptosis in hippocampal neurons of mice with cognitive impairment induced by sleep deprivation

  • Cao, Yin;Yang, Yingbo;Wu, Hui;Lu, Yi;Wu, Shuang;Liu, Lulu;Wang, Changhong;Huang, Fei;Shi, Hailian;Zhang, Beibei;Wu, Xiaojun;Wang, Zhengtao
    • Journal of Ginseng Research
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    • v.44 no.3
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    • pp.442-452
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    • 2020
  • Backgroud: Sleep deprivation (SD) impairs learning and memory by inhibiting hippocampal functioning at molecular and cellular levels. Abnormal autophagy and apoptosis are closely associated with neurodegeneration in the central nervous system. This study is aimed to explore the alleviative effect and the underlying molecular mechanism of stem-leaf saponins of Panax notoginseng (SLSP) on the abnormal neuronal autophagy and apoptosis in hippocampus of mice with impaired learning and memory induced by SD. Methods: Mouse spatial learning and memory were assessed by Morris water maze test. Neuronal morphological changes were observed by Nissl staining. Autophagosome formation was examined by transmission electron microscopy, immunofluorescent staining, acridine orange staining, and transient transfection of the tf-LC3 plasmid. Apoptotic event was analyzed by flow cytometry after PI/annexin V staining. The expression or activation of autophagy and apoptosis-related proteins were detected by Western blotting assay. Results: SLSP was shown to improve the spatial learning and memory of mice after SD for 48 h, accomanied with restrained excessive autophage and apoptosis, whereas enhanced activation of phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway in hippocampal neurons. Meanwhile, it improved the aberrant autophagy and apoptosis induced by rapamycin and re-activated phosphoinositide 3-kinase/Akt/mammalian target of rapamycin signaling transduction in HT-22 cells, a hippocampal neuronal cell line. Conclusion: SLSP could alleviate cognitive impairment induced by SD, which was achieved probably through suppressing the abnormal autophagy and apoptosis of hippocampal neurons. The findings may contribute to the clinical application of SLSP in the prevention or therapy of neurological disorders associated with SD.

Antioxidative and Anti-inflammatory Activity of Extract from Milling By-products of Sorghum Cultivar, 'Hwanggeumchal' (황금찰수수 도정부산물의 항산화 및 항염증 활성)

  • Ra, Ji-Eun;Park, Ji-Young;Seo, Woo Duck;Sim, Eun-Yeong;Ko, Jee Yeon;Nam, Min-Hee;Chung, Ill-Min;Han, Sang-Ik
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.4
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    • pp.463-469
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    • 2014
  • Sorghum has been consumed as one of the important staple food in the semiarid tropics of Africa and Asia. Sorghum is rich in starch, protein, essential vitamins and minerals and grows relatively well in dry climate regions when it compared with other staple food crops. Sorghum has taken an increased interest due to several studies that report about the beneficial effects of sorghum on human health. In the present study, we investigated the antioxidative and activity of extract of milling by-products (hull and bran) of Korean sorghum cultivar, 'Hwanggeaumchal' as well as its grain. Hull extract showed the highest total polyphenol contents ($29.7{\pm}0.2mg\;GAE/100g$) and major four pigments content ($322.6{\pm}14.5mg/100g$). From results of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate (ABTS) radical scavenging activity, hull extract ($IC_{50}$, $6.3{\pm}0.1{\mu}g/mL$) was also showed the strongest antioxidative effects. Bran and grain showed similar polyphenol, pigments contents and antioxidative effects. We determined cell viability by MTT assay and evaluated the anti-inflammatory activity by measuring nitric oxide (NO) of hull, bran and grain methanol extract (0.5% HCl v/v) on RAW 264.7 cells. Hull extract treatment was significantly decreased NO production with dose-dependant manner. Apigeninidin as one of the major pigment of hull was showed inhibitory activity against NO production without cytotoxicitiy. Therefore, sorghum milling by-products can be used as a good source of antioxidative and anti-inflammatory agents.