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Serum 25-Hydroxy Vitamin $D_3$ Analysis of Korean People (한국인 일반인의 혈청 25-Hydroxy Vitamin $D_3$의 분석)

  • Kim, Bo-Kyung;Jung, Hyun-Mi;Kim, Yun-Kyung;Kim, So-Young;Kim, Jee-Hyun
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.1
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    • pp.133-137
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    • 2010
  • Purpose: The main function of vitamin D is the mineralization of the brain by increase of calcium and phosphorus, in case it is insufficient in children, lime deposition on cartilage cannot occur so it leads to rachitis, and in adults, it leads to osteomalacia or osteoporosis. It is also strongly believed in the academic world that vitamin D can restrict the growth of cancer cells and prevent heart diseases, which is also somewhat proven in epidemiological researches. While the right density of vitamin D is still being studied, 20-32 ng/mL is believed to be the most ideal density. Therefore, I wanted analyze how much density of 25-Hydroxyvitamin D3 that Koreans possess. Materials and Methods: From February 20th, 2008 to April 21st, 2009, the collection of 2800 serums, from medical examination treated subjects by Neodin Medical Institute, have been tested. The targets were tested by 25-Hydroxyvitamin D (125I Kit: Diasorin, USA), and were analyzed by dividing into many different categories (gender, age, season, region). Results: The average density of male were 20 ng/mL, female 17.08 ng/mL. Per age groups, the density of males were as follows: 10~20-18 ng/mL, 21~30-17 ng/mL, 31~40-19 ng/mL, 41~50-21 ng/mL, 51~60-22 ng/mL, 61~70-22 ng/mL, 71~80-22 ng/mL and 81~90-19.9 ng/mL. Average density of females per age groups, were as follows: 10~20-16 ng/mL, 20~30-15.26 ng/mL, 30~40-16 ng/mL, 40~50-17 ng/mL, 50~60-19 ng/mL, 60~70-19 ng/mL, 70~80-19 ng/mL, and 80~90-17 ng/mL. Per seasons, From December to May, the subjects showed the density of 15.97 ng/mL, while from June to November, it showed 21.60 ng/mL. On density of males from January to April regionally, Seoul+Gyeonggi-Do-15.52 ng/mL, Gangwon-Do-15.33 ng/mL, Choongchung-Do-18.03 ng/mL, Jeonla-Do-18.68 ng/mL, Gyungsang-Do-18.76 ng/mL and Cheju Do-21.23 ng/mL. Conclusions: The vitamin D of Koreans is has been insufficient compared to the suggested amount. Ultraviolet rays, which is the main source of vitamin D is critical, therefore it is suggested that more outdoor activities can definitely help.

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Monitoring of Perfluorinated Compounds (PFCs) in the Seawater of the East and West Coast in Korea (동·서해안지역 해수에서 과불화 화합물의 모니터링)

  • Son, Jun-ho;Chung, Seon-Yong;Kwon, Bum Gun
    • Journal of the Korean GEO-environmental Society
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    • v.18 no.11
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    • pp.5-12
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    • 2017
  • Perfluorinated compounds (PFCs) as environmental pollutants are an important environmental issue. However, little is known on the PFCs monitoring of sea waters around estuaries on the East and West Coasts of Korea. This study shows the monitoring results of PFCs in sea waters in these Coasts. Among 10 PFCs selected in this study, concentrations of perfluorooctanesulfonate (PFOS), perfluorooctanoate (PFOA), perfluorobutane sulfonate (PFBS), and perfluorohexane sulfonate (PFHxS) in the East Coast were 9.6-50.7 ng/L (total mean${\pm}$standard deviation: $26.14{\pm}12.66ng/L$), 13.79-44.58 ng/L ($27.95{\pm}11.41ng/L$), limit of quantification (LOQ)-2.6 ng/L ($0.96{\pm}1.15ng/L$), and 2.95-11.05 ng/L ($4.25{\pm}2.57ng/L$), respectively. In the West Coast, concentrations of PFOS, PFOA, PFBS, and PFHxS were 27.66-51.71 ng/L ($36.27{\pm}7.79ng/L$), 8.97-22.53 ng/L ($14.47{\pm}4.25ng/L$), LOQ-2.27 ng/L ($1.63{\pm}0.93ng/L$), and 3.0-7.66 ng/L ($4.27{\pm}1.49ng/L$), respectly. Other PFCs were below LOQ. The result of this study provides the distribution pattern of PFCs for assessing environmental pollution in two coastal areas of Korea.

Analysis of Organotin Compounds in Fish and Shellfish from Guryongpo Harbor (구룡포항 어패류에 함유된 유기주석화합물 분석)

  • Kim, Mee-Ra;Kim, Sung-Hong
    • Korean Journal of Human Ecology
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    • v.14 no.1
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    • pp.209-216
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    • 2005
  • The concentrations of organotin compounds in fish and shellfish from Guryongpo harbor were analyzed. The concentrations of TBT, DBT and MBT in fish were in the range of $15.68{\sim}52.93\;ng/g$(average 28.65 ng/g), $13.95{\sim}72.16\;ng/g$(average 26.99 ng/g) and $37.73{\sim}133.59\;ng/g$(average 85.18 ng/g), respectively. The BT compound concentrations in intestine of fish were higher than those in muscle. Overall ranges of TBT, DBT and MBT concentrations in shellfish were $42.15{\sim}56.13\;ng/g$(average 50.94 ng/g), $34.16{\sim}52.82\;ng/g$(average 42.79 ng/g) and $41.25-72.60\;ng/g$(average 53.71 ng/g), respectively. The BT compound concentrations of shellfish were higher than those of fish. The concentrations of TPT and DPT in fish were $0.47{\sim}3.90\;ng/g$(average 2.33 ng/g) and $ND{\sim}0.56\;ng/g$(average 0.15 ng/g), but MPT was not detected. The concentrations of TPT, DPT and MPT in shellfish were in the range of $10.24{\sim}21.67\;ng/g$(average 17.45 ng/g), $3.32{\sim}4.78\;ng/g$(average 3.91 ng/g) and $0.11{\sim}0.36\;ng/g$(average 0.25 ng/g), respectively. In addition, the MBT ratio was highest among BT compounds, but the TPT ratio was the highest among PT compounds. Taken together, the concentrations of organotin compounds in fish and shellfish from Guryongpo harbor was not higher than those from other harbors in Korea.

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The Significance of Plasma Urokinase-type Plasminogen Activator and Type 1 Plasminogen Activator Inhibitor in Lung Cancer (폐암에서 혈장 Urokinase-Type Plasminogen Activator 및 Type 1 Plasminogen Activator Inhibitor의 의의)

  • Park, Kwang-Joo;Kim, Hyung-Jung;Ahn, Chul-Min;Lee, Doo-Yun;Chang, Joon;Kim, Sung-Kyu;Lee, Won-Young
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.516-524
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    • 1997
  • Background : Cancer invasion and metastasis require the dissolution of the extracellular matrix in which several proteolytic enzymes are involved. One of these enzymes is the urokinase-type plasminogen activator(u-PA), and plasminogen activator inhibitors(PAI-1, PAI-2) also have a possible role in cancer invasion and metastasis by protection of cancer itself from proteolysis by u-PA. It has been reported that the levels of u-PA and plasminogen activator inhibitors in various cancer tissues are significantly higher than those in normal tissues and have significant correlations with tumor size and lymph node involvement. Here, we measured the concentration of plasma u-PA and PAI-1 antigens in the patients with lung cancer and compared the concentration of them with histologic types and staging parameters. Methods : We measured the concentration of plasma u-PA and PAI-1 antigens using commercial ELISA kit in 37 lung cancer patients, 21 benign lung disease patients and 24 age-matched healthy controls, and we compared the concentration of them with histologic types and staging parameters in lung cancer patients. Results : The concentration of u-PA was $1.0{\pm}0.3ng/mL$ in controls, $1.0{\pm}0.3ng/mL$ in benign lung disease patients and $0.9{\pm}0.3ng/mL$ in lung cancer patients. The concentration of PAI-1 was $14.2{\pm}6.7ng/mL$ in controls, $14.9{\pm}6.3ng/mL$ in benign lung disease patients, and $22.1{\pm}9.8ng/mL$ in lung cancer patients. The concentration of PAI-1 in lung cancer patients was higher than those of benign lung disease patients and controls. The concentration of u-PA was $0.7{\pm}0.4ng/mL$ in squamous cell carcinoma, $0.8{\pm}0.3ng/mL$ in adenocarcinoma, 0.9ng/mL in large cell carcinoma, and $1.1{\pm}0.7ng/mL$ in small cell carcinoma. The concentration of PAI-1 was $22.3{\pm}7.2ng/mL$ in squamous cell carcinoma, $22.6{\pm}9.9ng/mL$ in adenocarcinoma, 42 ng/mL in large cell carcinoma, and $16.0{\pm}14.2ng/mL$ in small cell carcinoma. The concentration of u-PA was 0.74ng/mL in stage I, $1.2{\pm}0.6ng/mL$ in stage II, $0.7{\pm}0.4ng/mL$ in stage IIIA, $0.7{\pm}0.4ng/mL$ in stage IIIB, and $0.7{\pm}0.3ng/mL$ in stage IV. The concentration of PAI-1 was 21.8ng/mL in stage I, $22.7{\pm}8.7ng/mL$ in stage II, $18.4{\pm}4.9ng/mL$ in stage IIIA, $25.3{\pm}9.0ng/mL$ in stage IIIB, and $21.5{\pm}10.8ng/mL$ in stage IV. When we divided T stage into T1-3 and T4, the concentration of u-PA was $0.8{\pm}0.4ng/mL$ in T1-3 and $0.7{\pm}0.4ng/mL$ in T4, and the concentration of PAI-1 was $17.9{\pm}5.6ng/mL$ in T1-3 and $26.1{\pm}9.1ng/mL$ in T4. The concentration of PAI-1 in T4 was significantly higher than that in T1-3. The concentration of u-PA was $0.8{\pm}0.4ng/mL$ in M0 and $0.7{\pm}0.3ng/mL$ in M1, and the concentration of PAI-1 was $23.6{\pm}8.3ng/mL$ in M0 and $21.5{\pm}10.8ng/mL$ in M1. Conclusions : The plasma levels of PAI-1 in lung cancer were higher than benign lung disease and controls, and the plasma levels of PAI-1 in T4 were significantly higher than T1-3. These findings suggest involvement of PAI-1 with local invasion of lung cancer, but it should be confirmed by the data on comparison with pathological staging and tissue level in lung cancer.

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Research on Tongmu I Je-ma's medical idea (동무(東武) 이제마(李濟馬)의 의학사상(醫學思想)에 대한 연구(硏究))

  • Baek Sang-Yong
    • Journal of Korean Medical classics
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    • v.13 no.1
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    • pp.117-145
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    • 2000
  • Tongmu I Je-ma(1837-1900) set up Sasang Constitutional Medicine(四象體質醫學), a medical theory that would be original in the Korean medical history through his book ${\ll}$Tang-uisusebowon(東醫壽世保元)${\gg}$. In this book, he writes that he found the clue to his sasang Constitution(四象體質) theory from ${\ll}$Hwangjenaegyong(黃帝內經)${\gg}$. But the main logic that gives shape to the Constitution(體質) theory is $s{\check{o}}ng-ch{\check{o}}ng$ theory(性情論) of Confucian. Therefore, Tongmu(東武)'s Constitution $s{\check{o}}ng-ch{\check{o}}ng$ theory(體質性情論) is fundamentally based on confucian thoughts. Kongmaeng(孔孟) saw $s{\check{o}}ng$(性) as a base for connecting the entire mankind to be united, and $ch{\check{o}}ng$(情) as a individual thing that can exalted differently according to one's state of mind. Chuja(朱子) weakene the human's active volition by dividing $s{\check{o}}ng$ and $ch{\check{o}}ng$ as ibal(理發) and kibal(氣發) and therefore describing mankind to be submitted to Heaven's will. On the other hand Yolgok(栗谷) generalized $s{\check{o}}ng-ch{\check{o}}ng$ in the active mind of man himself by saying that $s{\check{o}}ng$ is when i-ki(理氣) does not act as a bodily function of the mind and $ch{\check{o}}ng$ is when it does. Furthennore he emphasized man's activeness on pursuing good by saying that the division of good and bad does not start from the origin of motion but is divided by the condition of ki(氣) which leads motion. Tongmu, who was influenced by Yulgok linked both $s{\check{o}}ng$and $ch{\check{o}}ng$ through, happiness, anger, sadness, pleasure(喜怒哀樂), and saw $s{\check{o}}ng$ as the ability to recognize good which is in the kijil(氣質) formed from the integration of i-ki, and saw $ch{\check{o}}ng$ as $s{\check{o}}ng$ blurred by man's greed. In addition to this, he says that the direct connection between each person's divergence in $s{\check{o}}ng-ch{\check{o}}ng$ and the condition of Four-organ(四臟) which is ki, not the mind which is i(理) forms sasangch'ejirin(四象體質人). His theory that illness comes from the partiality of $s{\check{o}}ng-ch{\check{o}}ng$ and therefore can prevent this by clarifying the mind and adjusting $s{\check{o}}ng-ch{\check{o}}ng$ through volition, has led Korean oriental medicine to be human based.

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The biblographical study on $T{\acute{o}}u\;f{\bar{e}}ng$ and Migraine -(Comparative study between Oriental and Western Medicine)- (두풍(頭風)과 편두통(Migraine)에 대(對)한 동서의학적(東西醫學的) 문헌고찰(文獻考察))

  • Oh, So-Jeo;Jeong, Ji-Cheon;Lee, Won-Chul
    • The Journal of Internal Korean Medicine
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    • v.14 no.1
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    • pp.129-138
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    • 1993
  • This report on the $T{\acute{o}}u\;f{\bar{e}}ng$ and Migraine comes to conclude, through the study of the Oriental- Western medical references, as follow; 1. First, $T{\acute{o}}u\;f{\bar{e}}ng$ and Migraine had some concurrencies that both the two symptoms have appeared severe and recurrent headache and more often to the female. 2 Many of them e.g. Sensory disturbance, Vertigo, Nausea, Vomiting, Tinnitus etc. in the prodrome and main symptom of $T{\acute{o}}u\;f{\bar{e}}ng$ and Migraine were identical, especially the symptom of the $f{\bar{e}}ng\;t{\acute{a}}n\;t{\acute{o}}u\;t{\grave{o}}ng$ was similar to the prodrome of the Migraine. We could find out the semilarity of the symptoms through that Migraine is proximately set in unilateral, and $Pi{\bar{a}}nT{\acute{o}}u\;f{\bar{e}}ng$ is so called alias $B{\grave{a}}n\;bi{\bar{a}}n\;t{\acute{o}}u\;t{\grave{o}}ng$. 3. The pathogeny of $T{\acute{o}}u\;f{\bar{e}}ng$ include the case of ‘$f{\bar{e}}ng\;xi{\acute{e}}\;r{\grave{u}}\;n{\bar{a}}o$’, the patient feeling weak condition, $T{\acute{a}}n,\;T{\acute{a}}nshi,\;T{\acute{a}}nhu{\breve{o}},\;Y{\grave{u}}q{\grave{i}}$, etc. and, ‘$t{\acute{a}}n\;zhu{\grave{o}}\;sh{\grave{a}}ng\;y{\acute{a}}o$’, ‘$G{\bar{a}}n\;y{\acute{a}}ng\;hu{\grave{a}}\;f{\bar{e}}ng$’. There were variable that $F{\bar{e}}ng,\;Xu{\grave{e}},\;F{\bar{e}}ngr{\grave{a}},\;F{\bar{e}}ngx{\bar{u}},\;Xu{\grave{e}}x{\bar{u}},\;Hu{\check{o}}$ in the left, and $t{\acute{a}}n,\;R{\grave{e}},\;t{\acute{a}}nr{\grave{e}},\;Qir{\acute{a}}$ in the right partial pathogeny. It was referred $Sh{\grave{a}}o\;y{\acute{a}}ng\;j{\bar{i}}ng$, $Ju{\acute{e}}\;y{\bar{i}}n\;j{\bar{i}}ng$, $Y{\acute{a}}ng\;m{\acute{i}}ng\;j{\bar{i}}ng$, $T{\grave{a}}i\;y{\acute{a}}ng\;j{\bar{i}}ng$ in connection with the Meridian system. And otherwise the primary cause of Migraine is still unknown to us. Heredity is probably important, but the mode of transmission is uncertain. Recently, the important assumption is the vasomotor change caused by vasoconstrictors like that norepinephrine, epinephrine, and serotonin etc.

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Study on Contration Distribution of HCB and DDTs in River Sediments of Korea (국내 주요 수계 표층 퇴적물 중 HCB와 DDTs의 농도분포 특성에 관한 연구)

  • Park, Jong-Eun;Lee, Sang-Chun;Hong, Jong-Ki;Kim, Jong-Guk
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.5
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    • pp.335-344
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    • 2012
  • Hexachlorobenzene (HCB) and Dichloro-Diphenyl-Trichloroethane (DDT) were determined in surface sediments collected from main rivers of Korea. Concentration of HCB in sediments ranged from 0.41 to 3.82 (average 1.58) ng/g, 0.08 to 6.09 (average 0.90) ng/g, 0.02 to 0.97 (average 0.30) ng/g, 0.28 to 0.59 (average 0.42) ng/g and 0.23 to 0.48 (average 0.32) ng/g in Han river, Nakdong river, Geum river, Yeongsan and Seomjin river respectively. The DDTs concentration was ranged from 0.67 to 14.20 (average 4.76) ng/g, N.D. to 10.36 (average 1.81) ng/g, N.D. to 7.26 (average 1.87) ng/g, N.D. to 3.12 (average 1.08) ng/g and 0.02 to 2.04 (average 0.56) ng/g in Han river, Nakdong river, Geum river, Yeongsan and Seomjin river respectively. In comparison with the concentration of HCB and DDTs in other studies, the values in sediments of this study were lower than those of other countries. Comparison with that Sediment quality guideline (SQG) of National Oceanic and Atmospheric Administration (NOAA), the HCB levels of this study were very lower than Effect Range Low (ERL) value. In the case of DDTs, the concentrations of 46 points were higher than ERL (1.58 ng/g). It have not harmful effect on ecosystem of the sediment, however ongoing monitoring of sediments is deemed necessary.

Concentrations of PM10 and Heavy Metals in Incinerators Around of Dong-hae Area (동해시 소각시설 주변의 입자상물질 및 중금속 농도현황에 관한 연구)

  • Son, Bu Soon;Lee, Jong Dae;Jung, Soon Won;Chung, Tae Woong
    • Journal of Environmental Impact Assessment
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    • v.14 no.5
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    • pp.355-363
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    • 2005
  • We studied the concentrations of $PM_{10}$ and heavy metal in the incinerators around Dong-hae from December 2003 to April 2004. The seasonal mean of $PM_{10}$ concentration was less than the value of environmental standard, the lowest ($36.59mg/m^3$) in spring and the highest ($78.54mg/m^3$) in fall. The measured seasonal heavy metals were Fe $503,44ng/m^3$, Mg $410.85ng/m^3$, Na $134.14ng/m^3$, Zn $24.58ng/m^3$ in spring and Na $315.29ng/m^3$, Mg $48.36ng/m^3$, Fe $44.78ng/m^3$, Zn $23.34ng/m^3$ in summer and Na $88.53ng/m^3$, Fe $33.80ng/m^3$, Zn $20,43ng/m^3$, Pb $6.16ng/m^3$ in fall and Na $4603.72ng/m^3$, Fe $575,26ng/m^3$, Al $411.01ng/m^3$, Cr $203.51ng/m^3$ in winter. The relationship between measured metals concentrations showed that Na and Fe were associated with natural sources such as soil. Pb, Cr and Zn were strongly related to urban anthropogenic sources such as fuel combustion.

Changes in Plasma Levels of Thyroid and Sex Steroid Hormones in Rockfish (Sebastes schlegeli) during Maturation and Parturition Periods (조피볼락(Sebastes schlegeli)의 성숙$\cdot$출산기에 있어서 갑상선 및 성 스테로이드 호르몬의 혈중 변화)

  • KANG Duk Yong;CHANG Young Jin;SOHN Young Chang;AIDA Katsumi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.4
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    • pp.574-580
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    • 1998
  • Changes in plasma levels of thyroid and sex steroid Hormones were examined during maturation and parturition periods in rockfish (Sebastes schlegeli) cultured in net pens. Plasma L-thyroxine levels were $35.2{\pm}5.7\;ng/m{\ell}\;(n=\;5;\;mean\;{\pm}\;sem)$ at vitellogenesis stage and then significantly decreased to $20.5\;{\pm}\;4.2\;ng/m{\ell}$ at parturition stage (P<0.05), and rapidly returned to high level, $44.9{\pm}\;7.2\;ng/m{\ell}$ at resting stage. Plasma 3,5,3'-triiodo-L-thyronine levels were 12.9 $\pm$ 1.6 ng/ml at vitellogenesis stage, but significantly decreased to $3.7{\pm}0.7\;ng/m{\ell}$ at ovulation stage (P<0.05) and increased to $52.9{\pm}7.0\;ng/m{\ell}$ at jesting stage. Plasma estradiol-17 $\beta$ level showed the highest value ($4.3{\pm}0.9\;ng/m{\ell}$) at vitellogenesis stage, but the level significantly decreased to $0.3{\pm}0.1\;ng/m{\ell}$, during parturition stage (P<0.05). In vitellogenesis and ovulation stages, plasma testosterone levels were $1.8{\pm}0.3\;ng/m{\ell}$ and $2.1{\pm}0.7\;ng/m{\ell}$, respectively, thereafter the level significantly decreased to $0.1{\pm}0.1\;ng/m{\ell}$ at parturition stage (P<0.05). These findings suggest that thyroid hormones may have relation to maturation and parturition of mother rockfish.

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THE EFFECT OF VITAMIN $D_3$ AND $TGF-{\beta}$ ON THE VIABILITY OF HUMAN PERIODONTAL LIGAMENT CELLS (Vitamin $D_3$$TGF-{\beta}$가 치주인대세포 활성에 미치는 영향에 관한 연구)

  • Song, Hyun-Sup;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.28 no.4 s.69
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    • pp.627-640
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    • 1998
  • [$TGF-{\beta}$] is a polypeptide with multiple physiological functions in regulation of cell-to-cell interaction and in growth and development. The active form of vitmain $D_3$, 1,25-dihydroxycholecalciferol $[1,25-(OH)_2D_3]$, is one of the most potent stimulators of osteoclastic acitivity. The purpose of this study was to evaluate the effect of Vitamin $D_3$ and/or $TGF-{\beta}$ on the periodontal ligament(PDL) cells. Human PDL cells were prepared from the first premolars extracted for the orthodontic treatment and were incubated in the environment of , $37^{\circ},\;5\%\;CO_2\;and\;95\%$ humidity. 10, 50 or 100ng/m1 of $1,25-(OH)_2D_3$ and 0.1, 1, 5 or 10ng/ml of $TGF-{\beta}$ were administered to the culture wells, separately or in combination. And the viability of PDL cells was evaluated by MTT assay The obtained results were as follows. 1. The viability of PDL cells in 10ng/ml of vitamin $D_3$ was not significantly differenent from that of the control group at 1, 2 and 3-day of cultivation, but it was significantly increased in 50ng/ml of Vitamin $D_3$ at 3-day and in 100ng/m1 of Vitamin $D_3$ at 2 and 3-day. 2. The viability of PDL cells in 0.1ng/ml of $TGF-{\beta}$ was not significantly differenent from that of the control group at 1, 2 and 3-day of cultivation, but it was significantly increased in 1 and 5ng/ml of $TGF-{\beta}$ at 3-day of cultivation, and in 10ng/ml of $TGF-{\beta}$ at 2 and 3-day of cultivation. 3. In case of admixture of 1ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$, the viability of PDL cells was significantly increased in the admixture of 100ng/ml of vitamin $D_3$ at 3-day of cultivation 4. In case of admixture of 5ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$, the viability of PDL cells began to be increased from 2-day of cultivation in the admixture of 10 50 and 100ng/ml of vitamin $D_3$, but it was not maintained at 3-day in the admixture of 10ng/m of vitamin $D_3$. 5. In case of admixture of 10ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$ the viability of PDL cells was significantly increased in the admixture of 50ng/ml of vitamin $D_3$ at 2 and 3-day of cultivation, and in the admixture of 100ng/ml at 1, 2 and 3-day.

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