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Clinical Analysis According to $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ Expression in Gastric Cancer (위암에서의 $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ 단백 발현)

  • Kim, Sin-Sun;Park, Yong-Geun;Jun, Kyong-Hwa;Jung, Hun;Song, Gyo-Young;Kim, Jin-Joo;Chin, Hyung-Min;Kim, Wook;Park, Cho-Hyun;Park, Seung-Man;Lim, Keun-Woo;Kim, Seung-Nam;Jeon, Hae-Myung
    • Journal of Gastric Cancer
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    • v.6 no.1
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    • pp.36-42
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    • 2006
  • Purpose: The $p21^{Waf1/Cip1}$ protein Inhibits the cell cycle by Inhibiting the phosphorylation at the $G1{\rightarrow}S$ check point, and the $p27^{kip1}$ protein similarly performs the suppressor function by controlling the p27-mediated G1 arrest. In this study, we analysed the clinical status and survival rates in correlations with p21 and p27 expression patterns in gastric cancer. Materials and Methods: Between 1993 and 1997, 192 patients who underwent surgeries in Catholic Medical Center were analysed retrospectively in this study. Immunohistochemical staining was performed and if the nuclei of the tumor cells were stained, we assumed those as positive results. Statistical analysis was based on clinicopathological findings and differences in survival rates. Results: The expression rate of p27 was 28.1% and 15.6% in p21 each. The ratio of T1-2(80.0%) was significantly high in p21 (+), but the ratio of T3-4 (50.6%) was slightly high in p21 (-). There was no statistical significance regarding other factors. The results in p27 was not much different from expression rate of p21 in T-stage. In addition, p27 expression in diffuse type (91.3%) was higher than in intestinal type (62.7%) by Lauren's classification (P<0.05). Also, there was no statistical significance in other factors. In the correlation of p21 and p27, p27 was positive when p21 was positive (53.5%). Conversely, p27 was negative when p21 was negative (76.5%, p<0.05). In the p21 and p27 combination test, there was higher rate of T1-2 (87.5%) in p21 (+)/p27 (+), and higher rate of T3-4 (58.1%) in p21 (-)/p27 (-) (P<0.05). Results showed higher rate of intestinal type (100%) in p21 (+)/p27 (+), and diffuse type (87.0%) was dominant in p21 (-)/p27 (-) (P<0.05) by Lauren's classification. Moreover, there was no statistical significance in the 5-year survival rate in the expression of p21 and p27, and the 5-year survival rate was highest in the case of p21 (+)/p27 (+) without statistical significance. Conclusion: In our study, $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ expressed similar patterns. The expression of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ affected the degree of invasiveness of the tumor, and. Combined examination result revealed the correlation of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ with Lauren's classification and depth of invasion of the tumor. However, we assumed that little difference between the survival rates depending on expression of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ has limited their value as predictable prognostic indicators.

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The Effect of Interferon-γ on Bleomycin Induced Pulmonary Fibrosis in the Rat (Interferon-γ 투여가 쥐에서의 Bleomycin 유도 폐 섬유화에 미치는 영향)

  • Yoon, Hyoung Kyu;Kim, Yong Hyun;Kwon, Soon Seog;Kim, Young Kyoon;Kim, Kwan Hyung;Moon, Hwa Sik;Park, Sung Hak;Song, Jeong Sup
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.1
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    • pp.51-66
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    • 2004
  • Objectives : The matrix metalloproteinases (MMPs) that participate in the extracellular matrix metabolism play a important role in the progression of pulmonary fibrosis. The effects of the MMPs are regulated by several factors including Th-1 cytokines, $interferon-{\gamma}$ ($IFN-{\gamma}$). Up to now, $IFN-{\gamma}$ is known to inhibit pulmonary fibrosis, but little is known regarding the exact effect of $IFN-{\gamma}$ on the regulation of the MMPs. This study investigated the effects of $interferon-{\gamma}$ on the pulmonary fibrosis and the expression of the lung MMP-2,-9, TIMP-1,-2, and Th-2 cytokines in aa rat model of bleomycin induced pulmonary fibrosis. Materials and methods : Male, specific pathogen-free Sprague-Dawley rats were subjected to an intratracheal bleomycin instillation. The rats were randomized to a saline control, a bleomycin treated, and a bleomycin+$IFN-{\gamma}$ treated group. The bleomycin+$IFN-{\gamma}$ treated group was subjected to an intramuscular injection of $IFN-{\gamma}$ for 14 days. At 3, 7, 14, and 28 days after the bleomycin instillation, the rats were sacrificed and the lungs were harvested. In order to evaluate the effects of the $IFN-{\gamma}$ on lung fibrosis and inflammation, the lung hydroxyproline content, inflammation and fibrosis score were measured. Western blotting, zymography and reverse zymography were performed at 3, 7, 14, 28 days after bleomycin instillation in order to evaluate the MMP-2,-9, and TIMP-1,-2 expression level. ELISA was performed to determine the IL-4 and IL-13 level in a lung homogenate. Results : 1. 7 days after bleomycin instillation, inflammatory changes were more severe in the bleomycin+$IFN-{\gamma}$ group than the bleomycin group (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$2.08{\pm}0.15:2.74{\pm}0.29$, P<0.05), but 28 days after bleomycin instillation, lung fibrosis was significantly reduced as a result of the $IFN-{\gamma}$ treatment (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$3.94{\pm}0.43:2.64{\pm}0.13$, P<0.05). 2. 28 days after bleomycin instillation, the lung hydroxyproline content was significantly reduced as a result of $IFN-{\gamma}$ treatment (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$294.04{\pm}31.73{\mu}g/g:194.92{\pm}15.51{\mu}g/g$, P<0.05). 3. Western blotting showed that the MMP-2 level was increased as a result of the bleomycin instillation and highest in the 14 days after bleomycin instillation. 4. In zymography, the active forms of MMP-2 were significantly increased as a result of the $IFN-{\gamma}$ treatment 3 days after the bleomycin instillation, bleomycin+$IFN-{\gamma}$ group (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$209.63{\pm}7.60%:407.66{\pm}85.34%$, P<0.05), but 14 days after the bleomycin instillation, the active forms of MMP-2 were significantly reduced as a result of the $IFN-{\gamma}$ treatment (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$159.36{\pm}20.93%:97.23{\pm}12.50%$, P<0.05). 5. The IL-4 levels were lower in the bleomycin and bleomycin+$IFN-{\gamma}$ groups but this was not significant, and the IL-13 levels showed no difference between the experiment groups. Conclusion : The author found that lung inflammation was increased in the early period but the pulmonary fibrosis was inhibited in the late stage as a result of $IFN-{\gamma}$. The inhibition of pulmonary fibrosis by $IFN-{\gamma}$ appeared to be associated with the inhibition of MMP-2 activation by $IFN-{\gamma}$. Further studies on the mechanism of the regulation of MMP-2 activation and the effects of MMP-2 activation on pulmonary fibrosis is warranted in the future.

Development for Fishing Gear and Method of the Non-Float Midwater Pair Trawl Net (II) - Opening Efficiency of the Model Net according to Front Weight and Wing-end Weight - (무부자 쌍끌이 중층망 어구어법의 개발 (II) - 추와 날개끝 추의 무게에 따른 모형어구의 전개성능 -)

  • 유제범;이주희;이춘우;권병국;김정문
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.39 no.3
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    • pp.189-196
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    • 2003
  • In this study, the vertical opening of the non-float midwater pair trawl net was maintained by controlling the length of upper warp. This was because the head rope was able to be kept linearly and the working depth was not nearly as changed with the variation of flow speed as former experiments in this series of studies have demonstrated. We confirmed that the opening efficiency of the non-float midwater pair trawl net was able to be developed according to the increase in front weight and wing-end weight. In this study, we described the opening efficiency of the non-float midwater pair trawl net according to the variation of front weight and wing-end weight obtained by model experiment in circulation water channel. We compared the opening efficiency of the proto type with that of the non-float type. The results obtained can be summarized as follows:1. The hydrodynamic resistance was almost increased linearly in proportion to the flow speed and was increased in accordance with the increase in front weight and wing-end weight. The increasing rate of hydrodynamic resistance was displayed as an increasing tendency in accordance with the increase in flow speed. 2. The net height of the non-float type was almost decreased linearly in accordance with the increase in flow speed. As the reduced rate of the net height of the non-float type was smaller than that of the net height of the proto type against increase of flow speed, the net height of the non-float type was bigger than that of the proto type over 4.0 knot. The net width of the non-float type was about 10 m bigger than that of the proto type and the change rate of net width varied by no more than 2 m according to the variation of the front weight and wing-end weight. 3. The mouth area of the non-float type was maximized at 1.75 ton of the front weight and 1.11 ton of the wing-end weight, and was smaller than that of the proto type at 2.0∼3.0 knot, but was bigger than that of the proto type at 4.0∼5.0 knot. 4. The filtering volume was maximized at 3.0 knot in the proto type and at 4.0 knot in the non-float type. The optimal front weight was 1.40 ton.

The Cox-Maze Procedure for Atrial Fibrillation Concomitant with Mitral Valve Disease (승모판막질환에 동반된 심방세동에서 Cox-Maze 술식)

  • Kim, Ki-Bong;Cho, Kwang-Ree;Ahn, Hyuk
    • Journal of Chest Surgery
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    • v.31 no.10
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    • pp.939-944
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    • 1998
  • Background: The sugical results of the Cox-Maze procedure (CMP) for lone atrial fibrillation(AF) have proven to be exellent. However, those for AF associated with mitral valve(MV) disease have been reported to be a little inferior. Materials and methods: To assess the efficacy and safety of the CMP as a combined procedure with MV operation, we studied retrospectively our experiences. Between April 1994 and October 1997, we experienced 70 (23 males, 47 females) cases of CMP concomitantly with MV operation. Results: The etiologies of MV disease were rheumatic in 67 and degenerative in 3 cases. The mean duration of AF before sugery was 66$\pm$70 months. Fifteen patients had the past medical history of thromboembolic complications, and left atrial thrombi were identified at operation in 24 patients. Twelve cases were reoperations. Aortic cross clamp (ACC) time was mean 151$\pm$44 minutes, and cardiopulmonary bypass (CPB) time was mean 246$\pm$65 minutes. Concomitant procedures were mitral valve replacement (MVR) in 19, MVR and aortic valve replacement (AVR) in 14, MVR and tricupid annuloplasty (TAP) in 8, MVR with AV repair in 3, MV repair in 11, MVR and coronary artery bypass grafting (CABG) in 2, MVR and AVR and CABG in 1, redo-MVR in 10, redo-MVR and redo-AVR in 2 patients. The rate of hospital mortality was 1.4%(1/70). Perioperative recurrence of AF was seen in 44(62.9%), and atrial tachyarrhythmias in 10(14.3%), low cardiac output syndrome in 4(5.7%), postoperative bleeding that required mediastinal exploration in 4(5.7%) patients. Other complications were acute renal failure in 2, aggravation of preoperative hemiplegia in 1, and transient delirium in 1 patient. We followed up all the survivors for 16.4 months(3-44months) on an average. Sinus rhythm has been restored in 65(94.2%) patients. AF has been controlled by operation alone in 73.9% and operation plus medication in 20.3%. Two patients needed permanent pacemaker implantation; one with sick sinus syndrome, and the other with tachycardia- bradycardia syndrome. Only two patients remained in AF. We followed up our patients with transthoracic echocardiography to assess the atrial contractilities and other cardiac functions. Right atrial contractility could be demonstrated in 92% and left atrial contractility in 53%.We compared our non-redo cases with redo cases. Although the duration of AF was significantly longer in redo cases, there was no differences in ACC time, CPB time, postoperative bleeding amount and sinus conversion rate. Conclusions: In conclusion, the CMP concomitant with MV operation demonstrated a high sinus conversion rate under the acceptable operative risk even in case of reoperation.

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Study on the Thermal Storage Characteristics of Phase Change Materials for Greenhouse Heating (온실보온(溫室保溫)을 위한 상변화(相變化) 물질(物質)의 축열특성연구(蓄熱特性硏究))

  • Song, Hyun-Kap;Ryou, Young-Sun;Kim, Young-Bok
    • Solar Energy
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    • v.13 no.2_3
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    • pp.65-78
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    • 1993
  • An overdose of fossil fuel for greenhouse heating causes not only the high cost and low quality of agricultural products, but also the environmental pollution of farm village. To solve these problems it is desirable to maximize the solar energy utilization for the heating of greenhouse in winter season. In this study phase change materials were selected to store solar energy concentratively for heating the greenhouse and their characteristics of thermal energy storage were analyzed. The results were summarized as follows. The organic $C_{28}H_{58}$, and the inorganic $CH_3COONa{\cdot}3H_2O\;and\;Na_2SO_4{\cdot}10H_2O$ were selected as low temperature latent heat storage materials. The equation of critical radius was derived to define the generating mechanism of the maximum latent heat of phase change materials. The melting point of $C_{28}H_{58}$ was $62^{\circ}C$, and the latent heat was $50.0{\sim}52.0kcal/kg$. The specific heat of liquid and solid phase was $0.54{\sim}0.69kcal/kg^{\circ}C$ and $0.57{\sim}0.75kcal/kg^{\circ}C$ respectively. The melting point of $CH_3COONa{\cdot}3H_2O$ was $61{\sim}62^{\circ}C$, the latent heat was $64.9{\sim}65.8$ kcal/kg and the specific heat of liquid and solid phase was respectively $0.83kcal/kg^{\circ}C$ and $0.51{\sim}0.52kcal/kg^{\circ}C$. The melting point of $Na_2SO_4{\cdot}10H_2O$ was $30{\sim}30.9^{\circ}C$, the latent heat was 53.0 kcal/kg and the specific heat of liquid and solid phase was respectively $0.78{\sim}0.89kcal/kg^{\circ}C$ and $0.50{\sim}0.7kcal/kg^{\circ}C$ When the urea of 21.85% was added to control the melting point of $Na_2SO_4{\cdot}10H_2O$ and the phase change cycles were repeated from 0 to 600, the melting point was $16.7{\sim}16.0^{\circ}C$ and the latent heat was $36.0{\sim}28.0kcal/kg^{\circ}C$.

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Assessment of the Usefulness of an IMRT Plan Using a Shell-Type Pseudo Target with Patients in Stage III or IV of NSCLC (비소세포폐암 III, IV기 환자에 있어서 Shell-Type Pseudo Target을 이용한 세기 조절 방사선치료계획기법의 유용성 평가)

  • Lee, Sang-Bong;Park, Ki-Ju;Park, Du-Chan;Kim, Man-Wo;Kim, Jun-Gon;Noh, Sung-Hwan
    • The Journal of Korean Society for Radiation Therapy
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    • v.24 no.2
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    • pp.95-106
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    • 2012
  • Purpose: The objective of this study was to investigate the usefulness of an IMRT treatment plan according to whether there was a shell-type pseudo target during radiation therapy for patients in Stage III or IV of non-small cell lung cancer (NSCLC). Materials and Methods: After setting an IMRT (Intensity-Modulated Radiation Therapy, IMRT) plan for when there was a shell-type pseudo target (SPT) and when there was none (WSPT) with 22 patients in Stage III or IV of NSCLC, the investigator analyzed dose-volume histograms (DVHs) and made assessment with dosimetric comparisons such as homogeneity index (HI) inside the tumor target, conformity index (CI) of the tumor target, spinal cord maximum dose, Esophagus $V_{50%}$, mean lung dose (MLD), and $V_{40%}$, $V_{30%}$, $V_{20%}$, $V_{10%}$, $V_{5%}$. Results: The mean CI of WSPT and SPT was $1.22{\pm}0.04$ and $1.16{\pm}0.032$ ($.000^*$), respectively, and the mean HI of WSPT and SPT was $1.06{\pm}0.015$ and $1.07{\pm}0.014$ ($.000^*$), respectively. In SPT, the mean of each CI difference decreased by $-5.16{\pm}2.54%$, while HI increased by average $0.81{\pm}0.47%$. Esophagus $V_{50%}$ recorded $14.54{\pm}12.01%$ (WSPT) and $12.14{\pm}11.09%$ ($.000^*$, SPT) with the mean of SPT differences dropping by $-26.37{\pm}25.05%$. Mean spinal cord maximum dose was $3,898.44{\pm}1,075.0$ cGy (WSPT) and $3,810.8{\pm}1,134.9$ cGy ($.004^*$, SPT) with SPT dropping by average $-3.36{\pm}5.81%$. As for lung $V_{X%}$, the mean of $V_{5%}$ and $V_{10%}$ differences was $-1.62{\pm}2.29%$ ($.006^*$) and $-1.98{\pm}5.02%$ ($.005^*$), respectively with SPT making a decrease. The mean of V20%, V30%, and V40% differences was $-3.51{\pm}3.07%$ ($.000^*$), $-4.84{\pm}6.01%$ ($.000^*$), and $-6.16{\pm}8.46%$ ($.001^*$), respectively, with SPT making a decrease with statistical significance. In MLD assessment, SPT also dropped by average $-2.83{\pm}2.41%$ ($.000^*$). Those results show that SPT allows for mean 169 cGy (Max: 547 cGy, Min: 6.4 cGy) prescription dose. Conclusion: An IMRT treatment plan with SPT during radiation therapy for patients in Stage III or IV of NSCLC will help to reduce the risk of lung toxicity and radiation-induced pneumonia by cutting down radiation doses entering the normal lung, reduce the local control failure rate during radiation therapy due to increasing prescription doses to a certain degree, and increase treatment effects.

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Bone Density and Related Factors of Food and Nutrition Major and Non-Major University Students in Seoul Area (서울지역 식품영양전공.비전공대학생의 골밀도에 미치는 영향요인에 관한 연구)

  • 정남용;최순남
    • Journal of the East Asian Society of Dietary Life
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    • v.13 no.5
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    • pp.391-407
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    • 2003
  • This study was conducted to investigate the factors affecting the born density of food and nutrition major and non-major university students in Seoul area. Data for food habits, dietary and health-related behavior were obtained by self administered questionnaires. BQI(bone quality index) of the subjects were measured by Quantitative Ultrasound (QUS). The results are summarized as follows: The average height, weight, BMI and osteopenia percentage of the major and non-major male and female student were l74.49cm, 67.05kg, 21.96 and 22.0%; l74.34cm, 65.98kg, 21.69 and 11.8%; l60.76cm, 54.48kg, 21.07 and 40.0%; l61.30cm, 54.22kg, 20.84 and 40.2%, respectively. The BQI of the major and non-major subjects were 108.07 and 110.47 in male student group, and 89.13, 88.18 in female student group, respectively. The T-score and Z-score of bone density of the subjects were not significantly different. Weight and BMI were positively related with BQI in male and female group but the relationship with BMI tended to be stronger in non-major female group than other groups. BQI was positively affected by exercise time, favorite food, and intake of seafood and tea in major and non-major male student group. One-side eating habit and intake of instant foods were negatively related with BQI in both male groups. In major and non-major female student group, exercise time, meal regularity, favorite food, amount of meal, intake of tofu were related with BQI positively and intake of tea and/or meats negatively. The result of this study revealed that desirable food habits, dietary behavior and health-related lifestyle may have a beneficial effect on bone density. They need practically and systematically organized nutrition education on optimum body weight, good eating habits, weight bearing exercise and intakes of good quality nutrient for higher bone density level.

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A Study on Garden Design Principles in "Sakuteiki(作庭記)" - Focused on the "Fungsu Theory"(風水論) - (「사쿠테이키(作庭記)」의 작정원리 연구 - 풍수론(風水論)을 중심으로 -)

  • Kim, Seung-Yoon
    • Journal of the Korean Institute of Landscape Architecture
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    • v.41 no.6
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    • pp.1-19
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    • 2013
  • This study tries to review 'Sakuteiki(作庭記)', the Book of Garden Making, compiled at the end of the 11th Century during the Heian Period of Japan, from the East-Asian perspective. 'Sakuteiki' is a Garden Theory Book, the oldest in the world as well as in Asia, and it contains the traditional knowledge of Japanese ancient garden culture, which originated from the continent(Korea and China). Traditional knowledge related to East-Asian garden culture reviewed in this paper is "Fungsu Theory"(風水, Asian traditional ecology: Fengshui in Chinese; Fusui in Japanese), stemmed from the culture to seek sound and blessed places to live in. Viewed from modern landscape architecture, the Fungsu Theory corresponds to ecology(science). The Fungsu Theory was established around the Han Dynasty of China together with the Yinyangwuxing(陰陽五行) Theory and widely used for making human residences including gardens. It was transmitted to Japan via Korea as well as through direct transaction between Japan and China. This study reinterprets garden design principles represented in Sakuteiki, which were selected in 5 key words according to the Fungsu Theory. The 5 key words for the Fungsu Theory are "the place in harmony of four guardian gods(四神相應地)", "planting trees in the four cardinal directions", "flow of Chi(氣)", "curved line and asymmetry", and "mountain is the king, water is the people". Garden design principles of "the place in harmony of four guardian gods(四神相應地)" and "planting trees in the four cardinal directions" are corresponding to "Myeongdang-ron(明堂論, Theory of propitious site)". The place in harmony of four guardian gods mentioned in Sakuteiki is a landform surrounded by the flow of water to the east, the great path to the west, the pond to the south, and the hill to the north. And the Theory originated from Zhaijing(宅經, Classic of dwelling Sites) of China. According to this principle, the city was planned and as a miniature model, the residence of the aristocrat during the Heian period was made. At the residence the location of the garden surrounded by the four gods(the flow of water, the great path, the pond, and the hill) is the Myeongdang(明堂, the propitious site: Mingtang in Chinese; Meido in Japanese). Sakuteiki explains how to substitute for the four gods by planting trees in the four cardinal directions when they were not given by nature. This way of planting originated from Zhaijing(宅經) and also goes back to Qiminyaoshu (齊民要術), compiled in the 6th Century of China. In this way of planting, the number of trees suggested in Sakuteiki is related to Hetu(河圖) and Luoshu(洛書), which are iconography of Yi(易), the philosophy of change, in ancient China. Such way of planting corresponds to that of Yongdoseo(龍圖墅, the villa based on the principle of Hetu) presented in Sanrimgyeongje (山林經濟), an encyclopedia on agriculture and living in the 17th Century of Korea. And garden design principles of "the flow of Chi(氣)", "curved line and asymmetry" is connected to "Saenggi Theory(生氣論, Theory of vitality)". Sakuteiki explains the right flow of Chi(氣) through the proper flow and the reverse flow of the garden stream and also suggests the curved line of the garden stream, asymmetric arrangement of bridges and stones in the garden, and indented shape of pond edges, which are ways of accumulating Chi(氣) and therefore lead to "Saenggi Theory" of the Fungsu Theory. The last design principle, "mountain is the king, water is the people", is related to "Hyeongguk Theory(形局論, Theory of form)" of the Fungsu Theory. Sakuteiki explains the meaning of garden through a metaphor, which views mountain as king, water as the people, and stones as king's retainers. It compares the situation in which the king governs the people with the help of his retainers to the ecological phenomena in which mountain(earth) controls water with the help of stones. This principle befits "Hyeongguk Theory(形局論, Theory of form)" of the Fungsu Theory which explains landform on the analogy of social systems, people, animals and things. As above, major garden design principles represented in Sakuteiki can be interpreted in the context of the Fungsu Theory, the traditional knowledge system in East Asia. Therefore, we can find the significance of Sakuteiki in that the wisdom of ancient garden culture in East-Asia was integrated in it, although it described the knowhow of a specific garden style in a specific period of Japan.

Effect on In­Vitro Fertilization of Pig Oocytes Matured in Different In­Vitro Maturation Media according to Sperm Concentration of Liquid Boar semen (돼지 액상정액의 정자 주입농도가 서로 다른 체외성숙배지에서 배양된 난포란의 체외수정에 미치는 영향)

  • 박창식;이영주;고현진;양창범;손동수;서길웅;이규승
    • Korean Journal of Animal Reproduction
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    • v.26 no.1
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    • pp.1-7
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    • 2002
  • The present study was carried out to investigate the effects of maturation media on penetrability of pig oocytes by liquid boar sperm coincubated with different sperm concentrations in a modified Tris­buffered medium (mTBM). Follicular oocytes collected from ovaries of prepubertal gilts were matured in a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. The sperm­ich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ far 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes in 5% $CO_2$in air at 38.5$^{\circ}C$, cumulus cells were removed and oocytes (30~40) were coincubated for 6 h in 0.5 $m\ell$ mTBM fertilization medium with five different (1$\times$10$^{6}$ , 2$\times$10$^{6}$ , 4$\times$10$^{6}$ , 6$\times$10$^{6}$, 10$\times$10$^{6}$ $m\ell$) sperm concentrations. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ NCSU-23 culture medium fur further culture of 6 h. At 12 h after IVF, sperm penetration, polyspermy and male pronuclear formation of oocytes were evaluated. Oocytes of NCSU-23 maturation medium decreased polyspermy and increased male pronuclear formation compared to those of mTCM­199 and mWaymouth MB 752/1 maturation media. Of oocytes matured in NCSU-23 medium and inseminated in mTBM medium with 2~4$\times$10$^{6}$ $m\ell$ sperm concentrations, 50.8~50.9% showed sperm penetration, 13.3~19.5% polyspermy and 43.9~45.4% male pronuclear formation. In conclusion, we found out that oocytes matured in NCSU­23 medium and inseminated in mTBM medium showed superior in­vitro fertilization compared to those matured in mTCM­199 and mWaymouth MB 752/1 maturation media and inseminated in mTBM medium. The optimum sperm concentrations for in-vitro fertilization of oocytes matured in NCSU-23 medium by liquid boar semen stored at 17$^{\circ}C$ for 5 days were 2~4$\times$10$^{6}$ $m\ell$.

In Vitro Fertilization of Pig Oocytes Matured In­Vitro by liquid Boar Spermatozoa (체외성숙 돼지 난포란의 액상정액을 이용한 체외수정)

  • 박창식;이영주
    • Korean Journal of Animal Reproduction
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    • v.26 no.1
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    • pp.17-23
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    • 2002
  • The present study was carried out to investigate the effects of the maturation media such as a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium on penetrability of pig oocytes by liquid boar sperm. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. When immature pig oocytes were cultured in mTCM-199, mWaymouth MB 752/1 and NCSU-23 maturation media for 44 h in 5% $CO_2$, in air at 38.5$^{\circ}C$, the germinal vesicle breakdown (CVBD) rates of the oocytes were 95.6, 94.1 and 94.9%, respectively, and the maturation rates (metaphase II) of oocytes were 92.5, 90.1 and 91.1%, respectively. No differences were observed among the maturation media. The spermrich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ for 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes, cumulus cells were removed and oocytes (30~40) coincubated far 6 h in 0.5 $m\ell$ mTCM-199 and mTBM fertilization media with 2$\times$1061$m\ell$ sperm concentration. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ mTCM-199 and NCSU-23 culture media for further culture 6 or 42 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF, and developmental ability of oocytes at 48 h after IVF were evaluated. The oocytes in combination with NCSU-23 medium for maturation and mTBM medium for IVF increased male pronuclear formation (48.0%) compared to those in combination with mTCM-199 media for maturation and IVF, and mWaymouth MB 752il medium for maturation and mTCM-199 medium far IVF. The rates of cleaved embryos (2~4 cell stage) at 48 h after IVF were 24.1% in combination with mTCM-199 media for maturation, IVF and culture, 43.6% in combination with mWaymouth MB 75211 medium fur maturation and mTCM-199 media for IVF and culture, and 71.2% in combination with NCSU-23 medium for maturation, mTBM medium for IVF and NCSU-23 medium for culture. In conclusion, we found out the oocytes matured in vitro were fertilized by liquid boar sperm stored in BTS extender at 17$^{\circ}C$ for 5 days. We recommend the simple defined NCSU-23 medium for nuclear maturation, mTBM medium and liquid boar sperm for IVF, and NCSU-23 medium for embryo culture.