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Apoptosis of Human Jurkat T Cells Induced by the Methylene Chloride Extract from the Stems of Zanthoxylum schinifolium is Associated with Intrinsic Mitochondria-Dependent Activation of Caspase Pathway (인체 급성백혈병 Jurkat T 세포에 있어서 Zanthoxylum schinifolium 줄기의 methylene chloride 추출물에 의해 유도되는 세포자살기전 규명)

  • Jun, Do-Youn;Woo, Mi-Hee;Park, Hae-Sun;Kim, Jun-Seok;Rhee, In-Koo;Kim, Young-Ho
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1499-1506
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    • 2008
  • To examine antitumor activity of the edible plant Zanthoxylum schinifolium, the cytotoxic effect of various organic solvent extracts of its stems on human acute leukemia Jurkat T cells was investigated. Among these extracts such as methanol extract (SS-7), methylene chloride extract (SS-8), ethyl acetate extract (SS-9), n-butanol extract (SS-10), and residual fraction (SL-11), SS-8 exhibited the most cytotoxic activity against Jurkat T cells. The methylene chloride extract (SS-8) possessed the apoptogenic activity capable of inducing sub-G1 peak along with apoptotic DNA fragmentation in Jurkat T cells. Western blot analysis revealed that SS-8 induced apoptosis via mitochondrial cytochrome c release into cytoplasm, subsequent activation of caspase-9 and caspase-3, and cleavage of PARP, which could be blocked by overexpression of Bcl-xL. Jurkat T cell clone I2.1 $FADD^{-/-}$) and Jurkat T cell clone I9.2 (caspase-$8^{-/-}$ were as sensitive as was the wild-type Jurkat T cell clone A3 to the cytotoxic effect of SS-8, suggesting no contribution of Fas/FasL system to the SS-8-mediated apoptosis. The GC-MS analysis of SS-8 showed that it was composed of 16 ingredients including 9,12-octadecanoic acid (18.62%), 2,4-dihydro-5-methyl-4- (1-methylethylidene)- 2-(4-nitrophenyl)-3H- pyrazol-3-one (14.97%), hexadecanoic acid (14.23%), (z,z)-6,9-pentadecadien- 1-ol (13.73%), 5,6-dimethoxy-2-methyl benzofuran (10.95%), and 4-methoxy-2-methylcinnamic acid (5.38%). These results demonstrate that the methylene chloride extract of the stems of Z. schinifolium can induce apoptotic cell death in Jurkat T cells via intrinsic mitochondria-dependent caspase cascade regulated by Bcl-xL without involvement of the Fas/FasL system.

Volatile Flavor Components and Free Radical Scavenging Activity of Cnidium officinale (천궁(Cnidium officinale)의 휘발성 향기성분 및 유리기 소거활성)

  • Lee, Ji-Hye;Choi, Hyang-Sook;Chung, Mi-Sook;Lee, Mie-Soon
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.330-338
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    • 2002
  • This study was performed to develop natural spices and functional foods using Cheongung (Cnidium officinale) which is one of the Korean medicinal plants. The volatile flavor patterns of Cnidium officinale were detected by electronic nose with 6 metal oxide sensors, and the principal component analysis was carried out. The volatile flavor components of Cnidium officinale were isolated by simultaneous steam-distillation extraction with pentane and diethylether (1 : 1), and essential oils were analyzed by capillary GC and GC/MS. The free radical scavenging activity of ethanol and methanol extracts from Cnidium officinale was measured by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and compared with ${\alpha}-tocopherol$ as reference. The principal component analysis showed the difference of principal components between fresh and drying samples. Eighty-five volatile flavor components (643.64 ppm) from fresh Cnidium officinale were identified and the major components were butyl phthalide, sabinene, neocnidilide. Sixty-four volatile flavor components (218.15 ppm) from hot air dried one were identified and the major components were butyl phthalide, sabinene, 3-N-butyl phthalide. And 73 volatile flavor components (784.15 ppm) from freeze dried one were identified and the major components were butyl phthalide, sabinene, ${\beta}-selinene$. The free radical scavenging activity of methanol cold extract (500 ppm) of freeze dried Cnidium officinale was higher than other samples. And methanol and ethanol cold extracts (above 250 ppm) of freeze dried sample were higher than ${\alpha}-tocopherol$ $25\;{\mu}M$ (22.34%).

Histological studies on in vitro Propagation of Pulsatilla koreana Nakai (할미꽃 기내증식(器內增殖)에 관(關)한 조직학적(組織學的) 연구(硏究))

  • Lee, Man-Sang;Oh, Ki-Hong
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.2
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    • pp.137-157
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    • 1993
  • This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

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Comparison of volatile flavor compounds of yuzu, kumquat, lemon and lime (유자, 금귤, 레몬 및 라임의 휘발성 향기성분의 비교)

  • Hong, Young Shin;Lee, Ym Shik;Kim, Kyong Su
    • Food Science and Preservation
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    • v.24 no.3
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    • pp.394-405
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    • 2017
  • This study was conducted to confirm the usefulness of essential oil components in yuzu and kumquat cultivated in Korea for comparison with those in lemon and lime. The volatile flavor compounds in citrus fruits (yuzu, kumquat, lemon and lime) were extracted for 3 h with 100 mL redistilled n-pentane/diethylether (1:1, v/v) mixture, using a simultaneous steam distillation and extraction apparatus (SDE). The volatile flavor compositions of the samples were analyzed by gas chromatography-mass spectrometry (GC-MS). The aroma compounds analyzed were 104 (3,713.02 mg/kg) in yuzu, 87 (621.71 mg/kg) in kumquat 103 (3,024.69 mg/kg) in lemon and 106 (2,209.16 mg/kg) in lime. Limonene was a major volatile flavor compound in four citrus fruits. The peak area of limonene was 35.03% in yuzu, 63.82% in kumquat, 40.35% in lemon, and 25.06% in lime. In addition to limonene, the major volatile flavor compounds were ${\gamma}$-terpinene, linalool, ${\beta}$-myrcene, (E)-${\beta}$-farnesene, ${\alpha}$-pinene and ${\beta}$-pinene in yuzu, and ${\beta}$-myrcene, ${\alpha}$-pinene, (Z)-limonene oxide, (E)-limonene oxide, geranyl acetate and limonen-10-yl acetate in kumquat. Furthermore, ${\gamma}$-terpinene, ${\beta}$-pinene, ${\beta}$-myrcene, geranyl acetate, neryl acetate and (Z)-${\beta}$-bisabolene in lemon and ${\gamma}$-terpinene, ${\beta}$-pinene, (Z)-${\beta}$-bisabolene, neral, geranial and neryl acetate in lime were also detected. As a result, it was confirmed that the composition of volatile flavor compounds in four citrus fruits was different. Also, yuzu and kumquat are judged to be worthy of use alternatives for lemon and lime widely used in the fragrance industry.

Determination of Hydrolysis Rate Constants on Phosphamidon and Profenofos (Phosphamidon과 Profenofos의 가수분해속도 상수의 측정)

  • Min, Kyung-Jin;Ha, Young-Duck;Seo, Seol;Cha, Chun-Geun;Park, Jang-Woo;Lee, Seung-Gon
    • Journal of Food Hygiene and Safety
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    • v.15 no.2
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    • pp.144-150
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    • 2000
  • The present study was peformed to determine the hydrolysis rate constants and degradation products of phosphamidon and proffnofos by the OECD method. Hydrolysis rate constants of phosphamidon in pH 4, pH 7, and pH 9 buffer solutions at 25 and 40$^{\circ}$C were 0.0020, 0.0022, 0.0049 and 0.0040, 0.0050, 0.0150, respectively. Hydrolysis rate of phosphamidon was accelerated by temprerature change under same pH conditions, and half-life of phosphamidon in pH 9 at 40。C was 3 times faster than that at 25。C. Hydrolysis rate of phosphamidon in alkaline solution(pH 9) was 2~4 times faster than that in acidic solution(pH 4) and neutral solution(pH 7) under same temperature. Hydrolysis rate constants of profenofos in pH 4, pH 7, and pH 9 buffer solutions at 25 and 40。C were 0.0022, 0.0047, 0.0860 and 0.0035, 0.0086, 0.1245, respectively. Hydrolysis rate of profenofos was accelerated by temprerature change under same pH conditions. Hydrolysis rate of profenofos in alkaline solution(pH 9) was 15~40 times faster than in acidic solution(pH 4) and neutral solution(pH 7) under same temperature condition, and half-life of profenofos was very fast within 8 hours. The hydrolysis rate of profenofos was faster than that of phosphamidon. In order to identify hydrolysis products, the extracts of degradation products were analyzed by GC/MS. The mass spectra of hydrolysis products of phosphamidon were at m/z 153 and 149, those of the profenofos were at m/z 208 and 240, respectively. The hydrolysis products of phosphamidon were O, O-dimethyl phosphate(DMP) and N, N-diethylchloroacetamide, and those of profenofos were 4-bromo-2-chlorophenol and O-ethyl-S-propyl phosphate.

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Changes in the inward current and membrane conductance after fertilization in the mouse eggs (수정에 의한 Mouse egg의 세포막전류 변화)

  • Hong, Seong-geun;Park, Choon-ok;Han, Jae-hee;Kim, Ik-hyun;Ha, Dae-sik;Kwun, Jong-kuk
    • Korean Journal of Veterinary Research
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    • v.32 no.2
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    • pp.157-164
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    • 1992
  • Changes in the both inward current and conductance of membrane by the fertilization were observed using the one microelectrode voltage clamp(or switch clamp) technique. Unfertilized eggs and both 1- and 2-cell stage eggs after fertilization were donated from the superovulated mouse (ICR, more than 6 weeks old) treated with PMSG(pregnant mare serum gonadotropin, Sigma) and HCG(human chorionic gonadotropin, Sigma) and naturally mated ones, respectively in this experiment. Membrane potential was held at -90mV and the voltage step was applied from -80mV to 50mV with interval of 10mV or 20mV for 300ms. since both of amplitudes and time courses in the membrane currents were various according to the states of cells and clamping condition, results were presented by their $averages{\pm}SEM$(standard mean error)and ratios or percentages. Inward currents began to appear in response to the step depolarization from -60mV and reached its maximum at -50mV. However, since the potential was not clamped evenly during the voltage step, current-voltage(I-V) relationship might be positively shifted 10 or 20mV. From the steady-state currents plotted in the I-V curve, outward rectification was markedly observed. Peak inward currents$(i_{in})$ at -50mV were $-0.62{\pm}0.23nA$(n=4),$-0.52{\pm}0.25nA$(n=5) and $-0.37{\pm}0.25nA$(n=6), in the 1-cell stage, 2-cell stage fertilized eggs and in the unfertilized eggs, respectively. Pure inward current (difference between steady-state and peak, $i_{in. pure}$) were $-1.01{\pm}0.23nA$, $-0.69{\pm}0.43nA$ and $-0.68{\pm}0.29nA$, respectively in the 1-cell stage fertilized eggs, unfertilized eggs and 2-cell stage fertilized eggs. These results suggested that the outward current in fertilized eggs of 2-cell stage was more increased than those in the unfertilized eggs. Pure inward currents in the all stages of eggs showed a similar fashion in the I-V relationship from -50mV to 50mV and reversal potential at 50mV. Time constant of inactivation$({\tau})$ in the inward current was decreased as the membrane potential was depolarized in the unfertilized and 2-cell stage eggs but in the 1-cell stage eggs t was not likely to be affected significantly. Slope conductances were 14.2nS, 8.9n5 and 7.7nS in the 1-cell, 2-cell stage fertilized eggs and the unfertilized eggs, respectively. Membranes between two cells within a zona pellucida seem to be electrical-connected in the 2-cell stage eggs from the observation made in the analysis for the electronic spread and decay to the current stimuli. Both of inward current and membrane conductance were increased after fertilization in the mouse eggs. Inward current seems to be carried by the same ion or through the same channels up to the 2-cell stage and ion that carried inward current was thought to play important function after fertilization in the mouse eggs.

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The Role of Adenosine Receptors on Acetylcholine Release in the Rat Striatum

  • Kim, Do-Kyung;Kim, Hyeon-A;Choi, Bong-Kyu
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.1
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    • pp.1-12
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    • 1997
  • As it has been reported that the depolarization induced acetylcholine (ACh) release is modulated by activation of presynaptic $A_1$ adenosine heteroreceptor and various evidence suggest that indicate the $A_2$ adenosine receptor is present in the striatum, this study was undertaken to delineate the role of adenosine receptors on the striatal ACh release. Slices from the rat striatum were equilibrated with $[^3H]$choline and then the release amount of the labelled product, $[^3H]$ACh, which was evoked by electrical stimulation (rectangular pulses, 3 Hz, 2 ms, 24 mA, $5\;Vcm^{-1}$, 2 min), was measured, and the influence of various agents on the evoked tritium outflow was investigated. And also, quantitative receptor autoradiography and drug-receptor binding assay were performed in order to confirm the presence and characteristics of $A_1$ and $A_2$ adenosine receptors in the rat striatum. Adenosine $(10{sim}100\;{mu}M)$ and $N^6$-cyclopentyladenosine (CPA, $1{sim}100\;{mu}M)$ decreased the $[^3H]$ACh release in a dose-dependent manner without changing the basal rate of release in the rat striatum. The reducing effects of ACh release by adenosine and CPA were abolished by 8-cyclopentyl-1,3-dipropy-Ixanthine (DPCPX, 2 ${mu}M$), a selective $A_1$, adenosine receptor antagonist, treatment. The effect of adenosine was potentiated markedly by 3,7-dimethyl-1-propargylxanthine (DMPX, 10 ${mu}M$), a specific $A_2$ adenosine receptor antagonist. 2-P-(2-carboxyethyl)phenethylamimo-5'-N- ethylcarboxamidoadenosine hydrochloride (CGS-21680C), in concentrations ranging from 0.01 to 10 ${mu}M$, a recently introduced potent $A_2$ adenosine receptor agonist, increased the $[^3H]$ACh release in a dose related fashion without changing the basal rate of release. These effects were completely abolished by DMPX $(10\;{mu}M)$. In autoradiograrhy experiments, $[^3H]$2-chloro-$N^6$-cyclopentyladenosine ($[^3H]$ CCPA) bindings were highly localized in the hippocampus and the cerebral cortex. Additionally, lower levels of binding were found in the striatum. However, $[^3H]$CGS-21680C bindings were highly localized in the striatal region with the greatest density of binding found in the caudate nucleus and putamen. Lower levels of binding were also found in the nucleus accumbens and olfactory tubercle. In drug-receptor binding assay, binding of $[^3H]$ CCPA to $A_1$ adenosine receptors of rat striatal membranes was inhibited by CPA ($K_i$ = 1.6 nM) and N-ethylcarboxamidoadenosine (NECA, $K_i$ = 12.9 nM), but not by CGS-21680C ($K_i$ = 2609.2 nM) and DMPX ($K_i$ = 19,386 nM). In contrast, $[^3H]$CGS-21680C binding to $A_2$ denosine receptors was inhibited by CGS-21680C ($K_i$ = 47.6 nM) and NECA ($K_i$ = 44.9 nM), but not by CPA ($K_i$ = 2099.2 nM) and DPCPX ($K_i$ = 19,207 nM). The results presented here suggest that both types of $A_1$ and $A_2$ adenosine heteroreceptors exist and play an important role in ACh release in the rat striatal cholinergic neurons.

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Effect of Lignans Isolated from Schisandra chinensis Baillon on Seed Germination and Seedling Growth in Radish (오미자 (Schisandra chinensis Baillon)로부터 순수분리한 리그난이 무의 종자발아 및 유묘생육에 미치는 영향)

  • Park, Da-Jung;Kim, Youn-Han;Park, Se-Jin;Rajasekar, Seetharaman;Park, Young-Hoon;Kang, Jum-Soon;Son, Beung-Gu;Lee, Yong-Jae;Kim, Sun-Tae;Yoon, Moo-Kyoung;Choi, Young-Whan
    • Journal of agriculture & life science
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    • v.46 no.1
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    • pp.91-103
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    • 2012
  • The aim of this study was to develop a rapid and cheap bioassay of four lignans (schisandrin, schisandrin C, the gomisin A and gomisin N) isolated from Schizandra chinensis Baill on seed germination and seedling growth of the radish. Its structure was determined by analysis of MS and NMR spectroscopic data. Radish seeds immersed for 1 hr in the solutions of $10^{-5}M$, $10^{-6}M$ and $10^{-7}M$ of schisandrin, schisandrin C, gomisin A, and gomisin N, seed germination was observed with in 60 hr after all of the treatments. Also, the seeds were germinated faster compared to untreated controls. At early germination stage, 48 hr after the treatment of the lignans, the suppression of germination was observed from all treatments; the suppression due to schisandrin and gomisin A was the highest at the concentration of $10^{-6}M$. The level of suppression increased as the concentration increased in the treatment of schisandrin C and gomisin N. Percent germination of seed after 184 hr was increased 90% at all treatments. For the effects of lignan treatment on seedling growth in radish, the growth of hypocotyl was promoted by gomisin A and gomisin N at all concentrations. Root elongation was significantly promoted by schisandrin and gomisin N at $10^{-5}M$ and $10^{-6}M$, respectively. Fresh and dry weights of the seedlings were high at a low concentration of $10^{-7}M$, but significantly reduced by schisandrin C at a high concentration of $10^{-5}M$. The results of the germination activity and seedling growth of the lignans from S. chinensis suggest their potential use as natural growth regulators.

Evaluation of Skin Dose and Image Quality on Cone Beam Computed Tomography (콘빔CT 촬영 시 mAs의 변화에 따른 피부선량과 영상 품질에 관한 평가)

  • Ahn, Jong-Ho;Hong, Chae-Seon;Kim, Jin-Man;Jang, Jun-Young
    • The Journal of Korean Society for Radiation Therapy
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    • v.20 no.1
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    • pp.17-23
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    • 2008
  • Purpose: Cone-beam CT using linear accelerator attached to on-board imager is a image guided therapy equipment. Because it is to check the patient's set-up error, correction, organ and target movement. but imaging dose should be cause of the secondary cancer when taking a image. The aim of this study is investigation of appropriate cone beam CT scan mode to compare and estimate the image quality and skin dose. Materials and Methods: Measurement by Thermoluminescence dosimeter (TLD-100, Harshaw) with using the Rando phantom are placed on each eight sites in seperately H&N, thoracic, abdominal section. each 4 methods of scan modes of are measured the for skin dose in three time. Subsequently, obtained average value. Following image quality QA protocol of equipment manufacturers using the catphan 504 phantom, image quality of each scan mode is compared and analyzed. Results: The results of the measured skin dose are described in here. The skin dose of Head & Neck are measured mode A: 8.96 cGy, mode B: 4.59 cGy, mode C: 3.46 cGy mode D: 1.76 cGy and thoracic mode A: 9.42 cGy, mode B: 4.58 cGy, mode C: 3.65 cGy, mode D: 1.85 cGy, and abdominal mode A: 9.97 cGy, mode B: 5.12 cGy, mode C: 4.03 cGy, mode D: 2.21 cGy. Approximately, dose of mode B are reduced 50%, mode C are reduced 60%, mode D are reduced 80% a point of reference dose of mode A. the results of analyzed HU reproducibility, low contrast resolution, spatial resolution (high contrast resolution), HU uniformity in evaluation item of image quality are within the tolerance value by recommended equipment manufacturer in all scan mode. Conclusion: Maintaining the image quality as well as reducing the image dose are very important in cone beam CT. In the result of this study, we are considered when to take mode A when interested in soft tissue. And we are considered to take mode D when interested in bone scan and we are considered to take mode B, C when standard scan. Increasing secondary cancer risk due to cone beam CT scan should be reduced by low mAs technique.

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Preliminary analysis of metabolic syndrome components in Korean adolescents by using Korean national health and nutrition examination Survey pooling data (1998, 2001, and 2005) (한국국민건강영양조사 병합자료(1998년, 2001년, 2005년)를 이용한 소아청소년에서의 대사증후군 진단 요인의 기초 분석)

  • Huh, Kyoung;Park, Mi Jung
    • Clinical and Experimental Pediatrics
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    • v.51 no.12
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    • pp.1300-1309
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    • 2008
  • Purpose :This study aimed to estimate age- and gender-specific cut points for metabolic syndrome (MS) components, including body mass index (BMI), blood pressure (BP), triglycerides, high-density lipoprotein (HDL) cholesterol, and glucose. Methods :Data from the 1998, 2001, and 2005 Korean NHANES (National Health and Nutrition Examination Survey) were analyzed (n=4164; 2,139 boys and 2,025 girls, aged 10-19 years). Height, weight, waist circumference (WC), BP, triglycerides, HDL cholesterol, and fasting glucose were measured. Results :BMI over $25kg/m^2$ represents the $85^{th}P$ (percentile) in 17-year-old boys and the $90^{th}P$ in 17-year-old girls. A level of WC higher than that of the cutoff points of Asian adults was found in the $90^{th}P$ of 17-year-old boys and girls. The $90^{th}P$ of boys aged 15 years old and the $95^{th}P$ of 13-year-old were included in the range of systolic BP over 130 mm Hg. Over the $75^{th}P$ of the group showed triglycerides greater than 110 mg/dL, (criterion of MS presented by NCEP-ATP III) and the $90^{th}P$ of the group showed triglycerides greater than 150 mg/dL by IDF. An HDL cholesterol level of 40 mg/dL represents the $25^{th}P$ in boys and the $10^{th}P$ in girls. A glucose level greater than 110 mg/dL represents the $95^{th}P$ and greater than 100 mg/dL represents the $90^{th}P$. Conclusion :Values of the $90^{th}P$ of MS components in late adolescent boys (WC, BP, and triglycerides) and girls (WC and triglycerides) were very high and in close proximity to the diagnostic criteria of adult MS.