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Inhibitory effects of calcium against intestinal cancer in human colon cancer cells and $Apc^{Min/+}$ mice

  • Ju, Jihyeung;Kwak, Youngeun;Hao, Xingpei;Yang, Chung S.
    • Nutrition Research and Practice
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    • v.6 no.5
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    • pp.396-404
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    • 2012
  • The aim of the study was to investigate the inhibitory effects of calcium against intestinal cancer in vitro and in vivo. We first investigated the effects of calcium treatment in HCT116 and HT29 human colon cancer cells. At the concentration range of 0.8-2.4 mM, calcium significantly inhibited cell growth (by 9-29%), attachment (by 12-26%), invasion (by 15-31%), and migration (by 19-61%). An immunofluorescence microscope analysis showed that the treatment with calcium (1.6 mM) for 24 h increased plasma membrane ${\beta}$-catenin but decreased nuclear ${\beta}$-catenin levels in HT29 cells. We then investigated the effect of dietary calcium on intestinal tumorigenesis in $Apc^{Min/+}$ mice. Mice received dietary treatment starting at 6 weeks of age for the consecutive 8 weeks. The basal control diet contained high-fat (20% mixed lipids by weight) and low-calcium (1.4 mg/g diet) to mimic the average Western diet, while the treatment diet contained an enriched level of calcium (5.2 mg calcium/g diet). The dietary calcium treatment decreased the total number of small intestinal tumors (by 31.4%; P < 0.05). The largest decrease was in tumors which were ${\geq}$ 2 mm in diameter, showing a 75.6% inhibition in the small intestinal tumor multiplicity (P < 0.001). Immunohistochemical analysis showed significantly reduced nuclear staining of ${\beta}$-catenin (expressed as nuclear positivity), but increased plasma membrane staining of ${\beta}$-catenin, in the adenomas from the calcium-treated groups in comparison to those from the control group (P < 0.001). These results demonstrate intestinal cancer inhibitory effects of calcium both in human colon cancer cells and $Apc^{Min/+}$ mice. The decreased ${\beta}$-catenin nuclear localization caused by the calcium treatment may contribute to the inhibitory action.

Identification and Functional Characterization of Novel Genetic Variations in the OCTN1 Promoter

  • Park, Hyo Jin;Choi, Ji Ha
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.2
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    • pp.169-175
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    • 2014
  • Human organic cation/carnitine transporter 1 (OCTN1) plays an important role in the transport of drugs and endogenous substances. It is known that a missense variant of OCTN1 is significantly associated with Crohn's disease susceptibility. This study was performed to identify genetic variants of the OCTN1 promoter in Korean individuals and to determine their functional effects. First, the promoter region of OCTN1 was directly sequenced using genomic DNA samples from 48 healthy Koreans. OCTN1 promoter activity was then measured using a luciferase reporter assay in HCT-116 cells. Seven variants of the OCTN1 promoter were identified, two of which were novel. There were also four major OCTN1 promoter haplotypes. Three haplotypes (H1, H3, and H4) showed decreased transcriptional activity, which was reduced by 22.9%, 23.0%, and 44.6%, respectively (p<0.001), compared with the reference haplotype (H2). Transcription factor binding site analyses and gel shift assays revealed that NF-Y could bind to the region containing g.-1875T>A, a variant present in H3, and that the binding affinity of NF-Y was higher for the g.-1875T allele than for the g.-1875A allele. NF-Y could also repress OCTN1 transcription. These data suggest that three OCTN1 promoter haplotypes could regulate OCTN1 transcription. To our knowledge, this is the first study to identify functional variants of the OCTN1 promoter.

Biological Activities of Sesquiterpene Lactones isolated from Several Compositae Plants Part 1 - Cytotoxicity against Cancer Cell Lines - (수종의 국화과 식물에서 분리한 Sesquiterpene Lactone들의 생리활성(제1보) - 암세포주에 대한 세포독성 -)

  • Jang, Dae-Sik;Park, Ki-Hun;Kim, Hwan-Mook;Hong, Dong-Ho;Chun, Hyo-Kon;Kho, Yung-Hee;Yang, Min-Suk
    • Korean Journal of Pharmacognosy
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    • v.29 no.3
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    • pp.243-247
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    • 1998
  • A diverse panel of human tumor cell lines and a mouse melanoma cell line (B16-F1) were used for the cytotoxicity test of the nine sesquiterpene lactones with ${\beta}-methylene-{\gamma}-lactone$ group isolated from Hemisteptia lyrata, Chrysanthemum zawadskii and Chrysanthemum boreale. In the cell adhesion inhibitory activity test against B16-F1 mouse melanoma cell, hemistepcin B, cumambrin B, costunolide and tulipinolide were shown significant activities with $IC_{50}$ range of 2.2, 4.1, 0.9 and $0.3\;{\mu}g/ml$, respectively. In the cytotoxicity test against human tumor cells, the most active compound was costunolide having $IC_{50}$ values of below $0.3\;{\mu}g/ml$ against all the tested cell lines except for UACC62. Cumambrin A, hendelin and costunolide exhibited more strong activity against HCT15 and UO-31 cell lines than a positive control, adriamycin. All tested compounds showed an $IC_{50}$ values of below $5.0\;{\mu}g/ml$ against all the tested cell lines.

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Phenolic Constituents from the Flowers of Hamamelis japonica Sieb. et Zucc.

  • Yim, Soon-Ho;Lee, Young Ju;Park, Ki Deok;Lee, Ik-Soo;Shin, Boo Ahn;Jung, Da-Woon;Williams, Darren R.;Kim, Hyun Jung
    • Natural Product Sciences
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    • v.21 no.3
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    • pp.162-169
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    • 2015
  • Hamamelis japonica (Hamamelidaceae), widely known as Japanese witch hazel, is a deciduous flowering shrub that produces compact clumps of yellow or orange-red flowers with long and thin petals. As a part of our ongoing search for phenolic constituents from this plant, eleven phenolic constituents including six flavonol glycosides, a chalcone glycoside, two coumaroyl flavonol glycosides and two galloylated compounds were isolated from the flowers. Their structures were elucidated as methyl gallate (1), myricitrin (2), hyperoside (3), isoquercitrin (4), quercitrin (5), spiraeoside (6), kaempferol 4'-O-β-glucopyranoside (7), chalcononaringenin 2'-O-β-glucopyranoside (8), trans-tiliroside (9), cis-tiliroside (10), and pentagalloyl-O-β-D-glucose (11), respectively. These structures of the compounds were identified on the basis of spectroscopic studies including the on-line LCNMR-MS and conventional NMR techniques. Particularly, directly coupled LC-NMR-MS afforded sufficient structural information rapidly to identify three flavonol glycosides (2 - 4) with the same molecular weight in an extract of Hamamelis japonica flowers without laborious fractionation and purification step. Cytotoxic effects of all the isolated phenolic compounds were evaluated on HCT116 human colon cancer cells, and pentagalloyl-O-β-D-glucose (11) was found to be significantly potent in inhibiting cancer cell growth.

The effect of Dochetang for the treatment of chronic diarrhea in a 4 months-old-female Korean native calf (만성설사를 보이는 생후 4개월령의 한우 암컷 송아지에서 도체탕의 치료효과)

  • Chon, Seung-Ki;Kim, Nam-Soo
    • Korean Journal of Veterinary Research
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    • v.47 no.2
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    • pp.233-239
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    • 2007
  • This study was carried out to evaluate the effects of Dochetang for the treatment of chronic diarrhea in a 4 months-old-female Korean native calf. The calf was presented to the Wow Animal Clinic, Iksan with the history of chronic diarrhea for several weeks. The feces test did not reveal the presence of the parasites or microbes causing diarrhea in calf. The blood test was also negative to the virus that causes of diarrhea in calf. Adminstration of parenteral antibiotics resulted in improvement of the condition temporarily but diarrhea was recurred again after 2-3 weeks. Then the calf was treated with Dochetang administered orally once a day in an empty stomach for 15 days. Feces was significantly reduced in moisture on 7 days after initial treatment. On 9 days after initial treatment, the calf had normal appetite and defecation in physiological conditions. Blood samples were collected before administration and on 1, 2 and 3 weeks after initial administration of Dochetang for hematology and biochemistry. A significantly differences were observed in the white blood cell (WBC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), albumin (ALB), glutamic pyrubic transaminase (GPT), inorganic phosphorus (IP) and magnesium (Mg), while no significant differences were seen in the red blood cell (RBC), hemoglobin (Hb), hematocrit (HCT), platelet (PLT), glucose (Glu), total protein (T-pro), glutamic oxaloacetic transaminase (GOT), blood urea nitrogen (BUN) and creatine (CRE). This study suggests that Dochetang administration can be a successful alternative therapeutic agent in instead of antibiotics for the treatment of chronic diarrhea in calves.

The Influences of Immune Function and Respiratory System on Aerobic Exercise by Exposed Acute Hypobaric Hypoxic Environment. (급성 저압.저산소 노출에서의 유산소성 운동이 면역기능 및 호흡기계에 미치는 영향)

  • Lee, Dong-Jun
    • Journal of Life Science
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    • v.19 no.6
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    • pp.825-831
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    • 2009
  • The purpose of this study was to investigate the influences of immune function and respiratory system on aerobic exercise of 75 %HRmax intensity in an exposed acute hypobaric hypoxic environment. The subjects were 20 male college students, 10 of which were soccer players (19.6${\pm}$0.8 yr, BMI 22.3${\pm}$1.0, V02max 56.5${\pm}$4.8 mljkgjmin) and 10 general students (22.2${\pm}$2.3 yr, BMI 22.7${\pm}$2.1, $VO_{2max}$ 50.5${\pm}$6.6 ml/kg/min). Items of measurement after exercise on level and high altitude were $O_{2}$, $CO_{2}$, ventilation (VE), respiratory frequency, respiratory quotient, RBC, MCV, MCH, MCHC, Hb, Hct, reticulocyte, WBC, neutrophil, lymphocyte, monocyte, basophil, and immunoglobulin (IgA, IgD, IgG, IgM). As the result of the verifying hypothesis, these results may suggest three new findings: first, to produce hypoxic ventilatory depression not only at level land but also at 3,000 m high altitude during 30 minute aerobics exercise of 75 %HRmax intensity, second to be more excellent for soccer players in adaptation and sensibility on response of respiratory system at high altitude, and third, to change each other by regular exercise habits or altitude in the monocyte.

A Study on the Dietary Behaviors and the Nutrient Intake of Middle School and High School Girls in Iksan (익산지역 여중생과 여고생의 식행동과 영양 섭취에 관한 연구)

  • Kim, In-Sook
    • Journal of the Korean Society of Food Culture
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    • v.17 no.5
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    • pp.560-568
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    • 2002
  • This study was carried out to compare the dietary behaviors and the nutrient intake of a middle school and a high school girls in Iksan. The subjects consisted of 132 middle school girls and 122 high school girls in Iksan. The proportion of subjects eating meals irregularly was higher in the high school girls (98.6%) than in the middle school girls(78.7%). The proportion of subjects skipping a meal also was higher in the high school girls(61.5%) than in the middle school girls(53.8%) and most of them (65.8%) skipped breakfast though they thought breakfast was the most important meal of a day (87.4%). The subjects tended to overeat at lunch (38.6%), and at dinner (55.9%). The main reasons of skipping a meal were 1. They had no time (49.3%), and 2. They had poor appetite (24.7%) Mean daily energy intake was 2198.5Kcal in the middle school girls and 2150.5Kcal in the high school girls and mean daily intake of protein, iron, vitamin $B_1$, vitamin $B_2$ and niacin was significantly higher (p<0.05, p<0.01) in the middle school girls than in the high school girls. Mean daily calcium, iron, vitamin $B_1$, vitamin A and C intake of middle school girls were lower than those of RDA and all nutrients intake of high school girls except for phosphorus was lower than those of RDA. The index of nutritional quality (INQ) was under 1.0 for calcium, iron, vitamin A, $B_1$ and C. Mean blood level of cholesterol, triglyceride, Hb, Hct, GOT, GPT were 170.9mg/dl, 78.9mg/dl, 13.1g/dl, 39.5%, 18.8mg/d and 15.7mg/dl, respectively.

Cytotoxic Triterpenoid from Rubus coreanus Miq

  • Lee, Dae-Young;Kim, Dong-Hyun;Bang, Myun-Ho;Song, Myoung-Chong;Kwak, Ho-Young;Yoo, Ki-Hyun;Chung, In-Sik;Kim, Kyong-Tai;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.275-280
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    • 2007
  • Dried unripe fruits of Rubus coreanus Miq. were extracted with 80% aqueous MeOH and the concentrated extract was partitioned with EtOAc and $H_2O$. From the EtOAc fraction, four triterpenoids were isolated through repeated silica gel, ODS and Sephadex LH-20 column chromatographies. From the result of physico-chemical data including NMR, MS aud IR, the chemical structures of the compounds were determined as tormentic acid (1), myrianthic acid (2), hovenic acid (3) and 2${\alpha}$,3${\beta}$,19${\beta}$,23-tetrahydroxylolean-12-en-28-oic acid (4). Compounds 3 and 4 were isolated for the first time from this plant. All isolated compounds were evaluated for cytotoxic activity against human colon carcinoma cells using in vitro three-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, compound 3 showed a higher cytotoxicity ($IC_{50}$ = 7.8 ${\mu}M$) than doxorubicin ($IC_{50}$= 50 ${\mu}M$).

Comparative Study on the Content and Cytotoxicity of Pseudolaric Acid B in the Five Plant Parts of Pseudolarix kaempferi

  • Nugroho, Agung;Woo, Nam-Tae;Park, Kyoung-Sik;Kwon, Na-Yun;Jung, Woo-Nyung;Lee, Sang Kook;Kim, Dong-Hwa;Park, Hee-Juhn
    • Natural Product Sciences
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    • v.23 no.4
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    • pp.265-269
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    • 2017
  • Pseudolaric acids of Pseudolarix kaempferi (Pinaceae) have been known as diterpenoids with potent anti-fungal-, anti-microbial, and cytotoxic activities. In the present study, the five MeOH extracts were prepared from the five plant part (root bark, stem bark, leaf, the inner part of root, and cone) to find the relation between the concentration of pseudolaric acids and cytotoxicity. Pseudolaric acids B and C were isolated from the root bark of P. kaempferi to use them as standard compounds. The five extracts were tested on cytotoxicity against six cancer cell lines, A549 (lung), HCT116 (colon), MDA-MB-231 (breast), SNU638 (stomach), and SK-hep-1 (liver) by SRB assay, but against K562 (leukemia) by SRB- or MTT assay. HPLC quantification were performed on a Shisheido Capcell PAK C18 column ($5{\mu}m$, $4.6mm{\times}250mm$) using 254 nm wavelength. The cytotoxicity ($IC_{50}$, $0.36{\mu}g/ml$ on K562 cell lines) of the root bark extract was potent and the content (101.1 mg/g extract) of pseudolaric acid B was very high in the root bark. These results suggest that the MeOH extract obtained from the root bark could be developed as the anti-cancer agent with a high quantity of pseudolaric acid B.

Phenolic Compounds from the Twigs of Corylopsis coreana Uyeki and Their Cytotoxic Activity (히어리나무의 페놀성 화합물 및 세포독성활성)

  • Kwon, Oh Kil;Kim, Chung Sub;Suh, Won Se;Park, Kyoung Jin;Cha, Joon Min;Choi, Sang Un;Kwon, Hak Cheol;Lee, Kang Ro
    • Korean Journal of Pharmacognosy
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    • v.47 no.1
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    • pp.1-6
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    • 2016
  • Phytochemical investigation of the twigs of Corylopsis coreana afforded 10 phenolic compounds, bergenin (1), 6'-O-galloylbergenin (2), 3'-O-galloylbergenin (3), (-)-catechin (4), (-)-epicatechin (5), (-)-epicatechin-3-O-galloyl ester (6), 4-methoxy-3,-5-dihydroxybenzoic acid (7), gallic acid (8), 2,4,6-trimethoxyphenol-1-O-${\beta}-\small{D}$-glucopyranoside (9), and 2,4,6-trimethoxyphenol-1-O-${\beta}-\small{D}$-(6-O-galloyl)-glucopyranoside (10). Their structures were characterized by spectroscopic data and identified by comparing these data with those in the literatures. The compounds 3, 9 and 10 were isolated for the first time from this source. All the isolates (1-10) were tested for their cytotoxic activity against A549, SK-OV-3, SK-MEL-2, and HCT15 cell lines in vitro using the SRB bioassay. The compounds 5, 7 and 8 exhibited selective cytotoxic activity against SK-MEL-2 cell line.