• Title/Summary/Keyword: College of Oriental Medicine

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Machilus Thunbergii Water Extract Induces Cytotoxic Effect against Human Acute Jurkat T Lymphoma (후박 열수 추출물의 Jurkat T 세포에서 세포사멸 효과)

  • Kim, Min Hwan;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.27 no.8
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    • pp.951-957
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    • 2017
  • To understand the cytotoxic activity of Machilus thunbergii, which has been used as a traditional oriental medicine, the mechanism underlying the cytotoxic effect of its extract on human acute Jurkat T cells was investigated. The methanol extract of roots (3 kg) of M. thunbergii was evaporated, dissolved in, and then extracted by water. The water-extracted active substance was designated MTWE. When Jurkat T cells were treated with MTWE at concentrations of 0, 25, 50, and $100{\mu}g/ml$, the apoptotic phenomenon of cells accompanying several subsequent biochemical reactions, such as mitochondrial cytochrome c release, activation of caspase-3, and ICAD degradation, was detected in the Jurkat T cells. Moverover. the expression of Bcl-xL, which is a suppressor for mitochondrial cytochrome c release pathway, was reduced in the Jurkat T cells. As DUSP6, a growth suppressor of cancer cells, ranged from 0, 25, 50, $100{\mu}g/ml$ of MTWE, the expression level was elevated in the Jurkat T cells. The apoptotic morphological change of the nuclei was observed by DAPI staining. Although the potential involvement of the other factors and DUSP6 is currently being investigated in more detail, these findings support the notion that MTWE is able to achieve the apoptosis of Jurkat T cells, and it seems that MTWE is useful as a method of evaluating a chemotherapeutic agent or tonic materials for human acute leukemia.

Systematic Identification of Hepatocellular Proteins Interacting with NS5A of the Hepatitis C Virus

  • Ahn, Ji-Won;Chung, Kyung-Sook;Kim, Dong-Uk;Won, Mi-Sun;Kim, Li-La;Kim, Kyung-Shin;Nam, Mi-Young;Choi, Shin-Jung;Kim, Hyoung-Chin;Yoon, Mi-Chung;Chae, Suhn-Kee;Hoe, Kwang-Lae
    • BMB Reports
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    • v.37 no.6
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    • pp.741-748
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    • 2004
  • The hepatitis C virus is associated with the development of liver cirrhosis and hepatocellular carcinomas. Among the 10 polyproteins produced by the virus, no function has been clearly assigned to the non-structural 5A (NS5A) protein. This study was designed to identify the hepatocellular proteins that interact with NS5A of the HCV. Yeast two-hybrid experiments were performed with a human liver cDNA prey-library, using five different NS5A derivatives as baits, the full-length NS5A (NS5A-F, amino acid (aa) 1~447) and its four different derivatives, denoted as NS5A-A (aa 1~150), -B (aa 1~300), -C (aa 300~447) and D (aa 150~447). NS5A-F, NS5A-B and NS5A-C gave two, two and 10 candidate clones, respectively, including an AHNAK-related protein, the secreted frizzled-related protein 4 (SFRP4), the N-myc downstream regulated gene 1 (NDRG1), the cellular retinoic acid binding protein 1 (CRABP-1), ferritin heavy chain (FTH1), translokin, tumor-associated calcium signal transducer 2 (TACSTD2), phosphatidylinositol 4-kinase (PI4K) and $centaurin{\delta}$ 2 ($CENT{\delta}2$). However, NS5A-A produced no candidates and NS5A-D was not suitable as bait due to transcriptional activity. Based on an in vitro binding assay, CRABP-1, PI4K, $CENT{\delta}2$ and two unknown fusion proteins with maltose binding protein (MBP), were confirmed to interact with the glutathione S-transferase (GST)/NS5A fusion protein. Furthermore, the interactions of CRABP-1, PI4K and $CENT{\delta}2$ were not related to the PXXP motif (class II), as judged by a domain analysis. While their biological relevance is under investigation, the results contribute to a better understanding of the possible role of NS5A in hepatocellular signaling pathways.

Involvement of Cdk Inhibitor p21(WIP1/CIP1) in G2/M Arrest of Human Myeloid Leukemia U937 Cells by N-Methyl-N'-Nitro-N-Nitrosoguanidine (N-methyl-N'-nitro-N-nitrosoguanidine에 의한 인체백혈병세포의 G2/M arrest 유발에서 Cdk inhibitor p21(WIP1/CIP1)의 관련성)

  • Choi, Yung-Hyun
    • Journal of Life Science
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    • v.19 no.1
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    • pp.1-8
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    • 2009
  • In this paper, to elucidate the further mechanisms of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced growth arrest, we investigated the effect of MNNG on cell cycle and proliferation in U937 cells, a p53-null human myeloid leukemia cell line. It was found that MNNG causes an arrest at the G2/M phase of the cell cycle and induces apoptosis, which is closely correlated to inhibition of cyclin B1 and cyelin-dependent kinase (Cdk) 2-associated kinase activities. MNNG treatment in. creased protein and mRNA levels of the Cdk inhibitor p21(WAF1/CIP1), and activated the reporter construct of a p21 promoter. By using p21 promoter deletion constructs, the MNNG-responsive element was mapped to a region between 113 and 61 relative to the transcription start site. These data indicate that in U937 cells MNNG can circumvent the loss of wild-type p53 function and induce critical downstream regulatory events leading to transcriptional activation of p21. Present results indicate that the p53-independent up-regulation of p21 by MNNG is likely responsible for the inhibition of cyclin/Cdk complex kinase activity rather than the down-regulation of cyclins and Cdks expression. These novel phenomena have not been previously described and provide important new insights into the possible biological effects of MNNG.

Isolation and Identification of Sesquiterpene o-Naphthoquinones, Mansonones E, F and H, from the Root Bark of Ulmus davidiana Planch (당느릅나무로부터 Sesquiterpene o-Naphthoquinone류 화합물, Mansonone E, F 및 H의 분리와 구조결정)

  • Kim, Jong-Pyung;Kim, Won-Gon;Koshino, Hiroyuki;Park, Jong-Hee;Jung, Jin;Yoo, Ick-Dong
    • Applied Biological Chemistry
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    • v.39 no.1
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    • pp.89-94
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    • 1996
  • Three sesquiterpene ortho-naphthoquinones were isolated from the methanolic extract of root bark of Ulmus davidiana Planch whose stem and root bark have been used as an oriental medicine for the treatment of edema, mastitis, gastric cancer and inflammation. The structures of these compounds were established on the basis of spectral data obtained from UV-vis, IR, HR-EIMS and NMR spectrometry, including the pulse field gradient (PFG)-HMQC and HMBC techniques. Their structures were determined as 2,3-dihydro-3,6,9-trimethylnaphtho(1,8-b,c)pyran-7,8-dione, 3,6,9-trimethylnaphtho(1,8-b,c)pyran-7,8-dione and 2,3-dihydro-4-hedroxy-3,6, 9-trimethylnaphtho(1,8-b,c)pyran-7,8-dione, which were identified as mansonones E. F and H, respectively. These compounds have originally been isolated from Mansonia altissima Chev, but have never been isolated from Ulmus davidiana Planch. Especially, mansonone H was isolated for the first time from Ulmaceae. The mismatched carbon chemical shifts of mansonones E and F in the reported literature were corrected by the aid of the PFG-HMBC spectral data.

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Effects of Radiation Mutant Perilla frutescens var. crispa in Preventing and Alleviating Symptoms in a Monosodium Iodoacetate-Induced Osteoarthritis Rat Model (MIA로 퇴행성관절염을 유도한 랫드에 방사선 형질전환 차조기가 증상 예방 및 완화에 미치는 효과)

  • Sim, Boo-Yong;Joo, In-Hwan;Kim, Sung-Kyu;Ji, Joong-Gu
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.4
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    • pp.830-838
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    • 2020
  • The present study aimed to evaluate the effect of radiation mutant Perilla frutescens var. crispa on bone metabolism and the inflammatory response in a monosodium iodoacetateinduced rat model of osteoarthritis. radiation mutant Perilla frutescens var. crispa was administered orally at doses of 25, 50 or 100 mg/kg/day for 2 weeks before direct injection of monosodium iodoacetate (3 mg/50 μl of 0.9% saline) into the intra-articular space of the rats' right knees. The rats subsequently received the same doses of oral radiation mutant Perilla frutescens var. crispa for another 4 weeks. It was evaluated that the treatment effects based on serum biomarkers, and morphological and histopathological analysis of the knee joints. Compared with those in control rats, the radiation mutant Perilla frutescens var. crispa treatments significantly reduced the serum levels of inflammation, bone metabolism markers (i.e., COX-2, LTB4, MMP-3, and COMP), and the amount of fibrous tissue. Otherwise, it was significantly increased the concentration of TIMP-1 and calcitonin. In addition, the radiation mutant Perilla frutescens var. crispa treatments effectively preserved the knee cartilage and synovial membrane. As a result, it indicates that radiation mutant Perilla frutescens var. crispa prevented and alleviated osteoarthritis symptoms. Thus, radiation mutant Perilla frutescens var. crispa can be used in food and drug material for the management of osteoarthritis.

Solid Cultivation of Fibrinolytic Enzyme (Bacillokinase) from Bacilis subtilis BK-17 (Bacillus subtilis BK-17 유래 혈전용해효소(Bacillokinase)의 고체배양)

  • Jeong, Yong-Kee;Beak, Hyun;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Joo, Woo-Hong;Kim, Jeong-In;Choi, Yung-Hyun;Chung, Kyung-Tae
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1478-1483
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    • 2009
  • A solid-state culture based on grain materials was attempted to produce a fibrinolytic enzyme for blood circulation improvement using Bacillus subtilis BK-17. The spore, rather than vegetative cell inoculation, of B. subtilis BK-17 on the solid-state culture was effective in the production of a fibrinolytic enzyme. Maximum spore production was obtained with a SFM medium (0.8% nutrient broth, 0.05% yeast extract, $10^{-1}$ M $MgCl_2$, $10^{-3}$ M $FeCl_3$, $10^{-4}$ $MnCl_2$, $10^{-5}$ M dipicolic acid, pH 6.5). Optimal pH and temperature were pH 6 and $30^{\circ}C$, respectively. The spore production reached a maximum at 60 hours of incubation. Bacillus subtilis BK-17 on the mung bean solid-state culture produced greater fibrinolytic activity, and less activity was seen in other grains such as kidney bean, soybean and corn. Protein and lipid contents of fermented soybeans were about 10 - 30% more than those of unfermented soybeans. Amino acid content was also 5 - 20% more than that of unfermented soybeans. These results indicated that fermented solid-state culture medium, fermented soybean in this case, can be utilized as a food supplement.

Piceatannol-Induced G1 Arrest of the Cell Cycle is Associated with Inhibition of Prostaglandin E2 Production in Human Gastric Cancer AGS Cells (Piceatannol에 의한 AGS 인체 위암세포의 G1 Arrest 및 Prostaglandin E2 생성의 억제)

  • Choi, Yung-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.7
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    • pp.907-913
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    • 2012
  • Piceatannol (trans-3,4,3',5'-tetrahydroxystilbene) is a polyphenol detected in grapes, rhubarb, and sugarcane. Although recent experimental data revealed that this compound is known to exhibit immunosuppressive and antitumorigenic activities in several cell lines, the molecular mechanisms underlying anticancer activity are poorly understood. In the present study, we investigated possible further mechanisms by which piceatannol exerts its anti-proliferative action in cultured human gastric cancer AGS cells. Piceatannol treatment resulted in the inhibition of growth and G1 arrest of the cell cycle in a concentration-dependent manner, as determined by MTT assay and flow cytometry analysis. The induction of G1 arrest by piceatannol was associated with the modulation of cyclin-dependent kinases (Cdks) and cyclins, up-regulation of the expression of Cdk inhibitor p21 (WAF1/CIP1) in both transcriptional and translational levels, and the inhibition of phosphorylation of retinoblastoma proteins and E2F1 expression. In addition, piceatannol treatment caused a progressive decrease in the expression levels of cyclooxygenase (COX)-2 without significant changes in the levels of COX-1, which was correlated with a decrease in prostaglandin $E_2$ synthesis.

Effect of Seeding Times on Yield and Flavonoid Contents of Mungbean (녹두 파종기에 따른 수량과 Flavonoid 함량 변화)

  • Kim, Dong-Kwan;Chon, Sang-Uk;Lee, Kyung-Dong;Kim, Kyong-Ho;Rim, Yo-Sup;Jeong, Seok-Cheol
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.3
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    • pp.273-278
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    • 2008
  • This study set out to investigate the changes to the growth, yield, and flavonoid contents of mungbean according to different seeding times from mid May to mid July in the southern region of Korea. Days to first flowering, days to first maturing, and cultivation period were shorter at later seeding time than earlier seeding time. But later the seeding time was, days from first maturing to first harvesting, days required between harvesting increased. Number of pods at first harvesting and yield of mungbean were highest when seeded at late June, showing increase in yield 14% more than at early June as standard seeding time. However number of seeds per pod and 1,000-seed weight at first harvesting were highest when seeded at mid July. The number of harvesting was smallest at two times when seeded at June or mid July. The contents of average vitexin and isovitexin in mungbean were highest in the order of mid July, late June, and mid May. In particular, their contents reached its highest point in the seeds of the second harvest. Considering the results of the cultivation period, yield, harvesting times, vitexin and isovitexin contents, the proper seeding time of mungbeans in the southern region of Korea will be from late June to mid July.

The Effect of Red Ginseng and Black Raspberry fermented by Lactobacillus acidophilus C in High Fat Diet-fed Mice (김치로부터 분리한 Lactobacillus acidophilus C로 발효한 홍삼 및 복분자 발효물의 비만예방 효과)

  • Moon, Hye-Jung;Lee, Joo-Hee;Lee, Sang-Cheon;Lee, Chang-Hyun;Choi, Kyung-Min;Cha, Jeong-Dan;Hwang, Seung-Mi;Ko, Eun-Sil;Lee, Min-Jung;Jang, Ji-Eun;Choi, Hye-Ran;Park, Jong-Hyuk
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.2
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    • pp.159-166
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    • 2015
  • We investigated the anti-obesity effects of Lactobacillus acidophilus C isolated from kimchi and red ginseng and black raspberry mixture fermented by this strain. Experimental samples were prepared with 10% skim milk, 1% red ginseng extract and 1% immature black raspberry powder and without red ginseng extract and black raspberry fermented by this strain at $37^{\circ}C$ for 24 hour. Male C57BL/6J mice were randomly divided into 4 groups (a normal diet group (N), a high-fat diet group (HF), a high fat diet with L. acidophilus C (C), and a high fat diet with fermented mixture (CEx)) and were orally administered daily for 6 weeks at concentration of 2,000 mg/kg body weight. The results showed that weight gain, weight of periepididymal fat and perirenal fat, adipocyte size, lipid accumulation in liver, serum glucose concentration, serum GOT and GPT concentration and serum leptin were decreased and concentration of HDL-cholesterol and serum adiponectin were increased compared to HF group. These results suggest that fermented mixture (CEx) might be helpful in improvement of high-fat diet-induced obesity than only L. acidophilus C intake.

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Purification and Biochemical Characteristics of Fibrinolytic Enzyme from Streptomyces corcohrussi JK-20 (Streptomyces corcohrussi JK-20 유래 혈전용해효소의 순수분리 및 이의 생화학적 특성 규명)

  • Kim, You-Jung;Park, Jeong-Uck;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Jin, Se-Hun;Kang, Byoung-Won;Choi, Yung-Hyun;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.20 no.6
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    • pp.838-844
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    • 2010
  • A fibrinolytic enzyme of Streptomyces corcohrussi from soil sediment was purified by chromatography using DEAE-Sephadex A-50 and Sephadex G-50. The analysis of SDS-polyacrylamide gel suggested that the purified enzyme is a homogeneous protein and the molecular mass is approximately 34 kDa. The purified enzyme showed activity of 0.8 U/ml in a plasminogen-rich fibrin plate, while its activity in a plasminogen-free fibrin plate was only 0.36 U/ml. These results suggested that the purified enzyme acts as a plasminogen activator. The fibrinolytic activity of the enzyme under the supplementation of protease inhibitors, $\varepsilon$-ACA, t-AMCHA and mercuric chloride in the enzyme reaction was less than 24%, indicating that it could be modulated by the plasmin and/or fibrinogen inhibitors involved in the fibrinogen-to-fibrin converting process. As time passed, $Zn^{2+}$, a heavy metal ion, inhibited the activity to 34.1%. The optimum temperature of the purified enzyme was approximately $50^{\circ}C$ and over 92% of the enzyme activity was maintained between pH 5.0 and 8.0. Therefore, our results provide a potential fibrinolytic enzyme as a noble thrombolytic agent from S. corcohrussi.