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A Reliable Protocol for transfection of mature primary hippocampal neurons using a neuron-glia co-culture system (신경세포-신경교세포 공동배양을 이용한 성숙한 해마신경세포의 효율적인 형질전환 방법)

  • Lee, Hyun-Sook;Cho, Sun-Jung;Jung, Yong-Wook;Jin, Ing-Nyol;Moon, Il-Soo
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.198-203
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    • 2007
  • DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.

Needs on Management Development Program for Head Nurse (간호 관리 능력 개발을 위한 교육 연구)

  • Park, Jeong-Sun
    • Journal of Home Health Care Nursing
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    • v.5
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    • pp.84-99
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    • 1998
  • The objectives of this study are to identify the actual educational contents of management for head nurse and to propose the educational subjects according to identity the needs of head, charge, and staff nurses. The subjects were investigated the actual Management Development Programs and educational needs of head nurses and prospective nurse manager(charge nurse, staff nurse with a lot of clinical experiences) in general hospitals. The tools were composed of two questionnaires: One was developed from the literature review for making items to measure actual situation. The other was revised Katz's model for measurement of educational needs. The first respondents of actual situation were 27 general hospitals with over 400 beds in Seoul and the second respondents were 89 head nurses, 67 charge nurses and 136 nurses at 3 hospitals by convenient sampling out of 27 general hospitals. Data were collected by telephone interview, mail questionnaire and visiting from 7th of October through 30th of November in 1997. In data analysis, general characteristics of the respondents and actual status of Management Development Programs were analyzed by frequency and percentage. Educational needs according to general characteristics were analyzed by ANOVA The results were as follows: 1. Actual situation of Management Development Program 1) Seven hospitals(26%) had Management Development Program for prospective managers and 14 hospitals (52%) for head nurses. 2) Education Department existed in 14 hospitals (52%). 3) One hospital(4%) had top level managers took part in the Management Development. 4) Two hospitals selected head nurse, who had finished courses of Management Development. Eight hospitals(30%) assessed educational needs. The assessment tools consisted of making a question via questionnaire(75%), determining at department meeting(12%) and interview(13%). 5) Educational programs had 3 types: 10 lecture type, 7 discussion type and 4 role play type programs. 6) One hospital evaluated the change of learner's attitude. 7) Four hospitals scored educational point, but that was measured only by attending. 8) Actual Management Development Programs were as follows. parenthesis indicates the number of hospitals. (1) Management Development Programs for Prospective manager. Role perception of Middle level Manager (1) . Role reconstruction of Nurse Manager (1). Workshop for Charge Nurse (1). Nursing Delivery System and Nursing Process (1). Communication (1). Motivation (1) (2) Management Development Programs for Head nurse.. Head nurse's Role (5). Administrative Work (7). Service Education (4). Prevention and Countermeasure of Nursing Incidence (3). Appraisal (3) 2. The results of needs on Management Development subject 1) The educational needs of all respondents on 3 skill domains showed positive agreement to strongly positive agreement. 2) High priority(more than 4.5) items were 12 of 24 Human skill items(50%), 1 of 6 Technical skill items(16%), and 2 of 13 Conceptual skill items (15%). 3) Out of high priority items, 8 items were instituted. 4) All respondents showed high needs on 3 skill domains regardless of 3 positions (head nurse, charge nurse, and nurse). Educational needs of Human skill domain, according to position were 108. S, 108.7, 106.8 (mean score = 72) , needs of Technical skill domain were 26.5, 26.6, 26.I(mean score=18), and needs of Conceptual skill domains were 56.9,56.7, 55.1(mean score=39). 5) Needs of 3 skill domains according to clinical career showed significant difference. Out of respondents, nurses with career of over 16years showed lowest degree of needs in Human skill domains(F=4.47, P=.004) and Conceptual skill domain(F=2.93, P=.034). 6) Educational needs according to educational background were not significant difference. But out of respondents, nurses educated at 3-year junior college relatively showed lowest needs in all of the 3 skill domains. With the above-mentioned findings, further study is necessary for generalization of this study at hospitals with different bed size and location. Also it is needed to study about management skill of nurse and charge nurse, and effective educational method.

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Review of the Radiation Risk and Clinical Efficacy Associated with Computed Tomography Cancer Screening (암의 조기발견을 위한 CT촬영에서의 임상적 효능과 방사선위해에 대한 고찰)

  • Kim, Hyun Ja
    • Journal of Radiation Protection and Research
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    • v.38 no.4
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    • pp.214-227
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    • 2013
  • Computed tomographic scan as a screening procedures in asymptomatic individuals has seen a steady increase with the introduction of multiple-raw detector CT scanners. This report provides a brief review of the current controversy surrounding CT cancer screening, with a focus on the radiation induced cancer risks and clinical efficacy. 1. A large study of patients at high risk of lung cancer(the National Lung Screening Trial[NLST]) showed that CT screening reduced cancer deaths by 20%(1.33% in those screened compared with 1.67% in those not screened). The rate of positive screening tests was 24.2% and 96.4% of the positive screening results in the low-dose CT group were false-positive. Radiation induced lung cancer risk was estimated the most important in screening population because ERR of radiation induced lung cancer does not show the decrease with increasing age and synergistic connection between smoking and radiation risk. Therefore, the radiation risk may be on the same order of magnitude as the benefit observed in the NLST. Optimal screening strategy remain uncertain, CT lung cancer screening is not yet ready for implementation. 2. Computed tomographic colonography is as good as colonoscopy for detecting colon cancer and is almost as good as colonoscopy for detecting advanced adenomas, but significantly less sensitive and specific for smaller lesions and disadvantageous for subsequent therapeutic optical colonoscopy if polyps are detected. The average effective dose from CT colonography was estimated 8-10 $mS{\nu}$, which could be a significant dose if administered routinely within the population over many years. CT colonography should a) achieve at least 90% sensitivity and specificity in the size category from 6 and 10 mm, b) offer non-cathartic bowl preparation and c) be optimized and standardized CT parameters if it is to be used for mass screening. 3. There is little evidence that demonstrates, for whole-body scanning, the benefit outweighs the detriment. This test found large portion of patient(86~90.8%) had at least one abnormal finding, whereas only 2% were estimated to have clinically significant disease. Annual scans from ages 45 to 75 years would accrue an estimated lifetime cancer mortality risk of 1.9%. There is no group within the medical community that recommends whole-body CT. No good studies indicate the accuracy of screening CT, at this time. The benefit/risk balance for any of the commonly suggested CT screening techniques has yet to be established. These areas need further research. Therefore wild screening should be avoided.

Bronchoscopic Electrocautery with Electroprobe and Diathermic Snare in Patients with Malignant Airway Obstruction (중심성 폐암환자에서 악성 기도폐쇄에서 전기 탐침과 투열올가미를 사용한 전기 소작술의 효과)

  • You, Hong Jun;Choi, Pyoung Rak;Park, Eun Ho;Yang, Jae Hong;Choi, In Su;Kim, Hyoung Jun;Kim, Joo Hoon;Oak, Chul Ho;Cho, Hyun Myung;Jang, Tae Won;Jung, Maan Hong
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.5
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    • pp.504-509
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    • 2005
  • Background : Endobronchial tumors cause life-threatening dyspnea and can lower the quality of life due to central airway obstruction. In those cases with an intraluminal tumor, various bronchoscopic techniques are available for tumor debulking. The therapeutic effect of bronchoscopic electrocautery for palliation in patients with a symptomatic tumor obstruction was studied. Method : Nineteen patients with bronchogenic carcinomas (n=15) and metastatic tumors affecting the bronchi (n=4), between March 2002 and March 2005, were enrolled in this study. Electrocautery was performed under local anesthesia using an electroprobe and diathermic snare. Using flexible bronchoscopy, a follow-up bronchoscopic examination was performed 3-4 days later. Symptom improvement was evaluated by $FEV_1$, FVC and dyspnea score (Modified Borg Category Scale (0~10)), both before and after the electrocautery. Results : The success rate of electrocautery on the follow up examination was 84%. Patients with endoluminal airway lesions had a mean overall decrease in the size of the obstruction to $47.8{\pm}15.7%$. The mean Improvement in the dyspnea score immediately after the endobronchial tumor debulking was $2.78{\pm}1.42$. The average improvements in the $FEV_1$ and FVC after electrocautery were $0.32{\pm}0.19L$ and $0.5{\pm}0.22L$, respectively. There were 2 cases of complications related with electrocautery (one each of pneumothorax and pneumonia). Conclusion : Electrocautery using an electroprobe and diathermic snare was an effective and safe palliative treatment for a symptomatic endoluminal airway obstruction in lung cancer.

Changes of Qualities of Green Asparagus Packed with Different Types during Low Temperature Storage (포장 방법에 따른 아스파라거스의 저온저장 중 품질변화)

  • Wang, Lixia;Choi, In-Lee;Kang, Ho-Min
    • Journal of Bio-Environment Control
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    • v.29 no.3
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    • pp.239-244
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    • 2020
  • Effect of 6kg large unit with the carton box (20% open ratio) and MA box (10,000cc·m-2·day-1·atm-1 oxygen transmission rates modified atmosphere package), and the 100g small unit with MA film on asparagus sensory quality were evaluated. The CO2 concentration depended largely on the packing unit and maintained at around 3% in small MA packages, whereas in the MA box increased to 12%. Ethylene concentration rapidly increased until after 3 days of storage in MA packages and then decreased to maintain 5μL·L-1. Unrelated to the unit size, the lower weight loss was obtained in MA packages. A significant difference in visual quality was shown since the 15th day, the best and worst were the MA box and small MA package on the finish day. Off-odor was the highest in small MA packages and the lowest in the carton box (< 3.0). Although there was no significant difference in firmness among all treatments, the packages showed the highest firmness in tips and stems, respectively. The sugar content and hue angle decreased during storage, but there was no statistical difference in all treatments. EL was lowest and highest in small MA package and carton box, respectively. On the 10th day, the total aerobic bacteria was lowest in small MA packages, but no significant difference on the 20th day. E. coli was not found in all treatments on the 10th day, while it was the lowest in the MA box on the 20th day. The mold and yeast were not observed during the whole storage. Based on the above results, the carton box packaged with 10,000cc OTR film was more effective in maintaining the quality of green asparagus with the suitable CO2 concentration for asparagus cold storage.

Characteristics of $Malassezia$ $pachydermatis$ Isolated from Dogs and Antifungal Effect of Essential Oils (개에서 분리된 $Malassezia$ $pachydermatis$의 특성과 Essential Oil의 항진균 효과)

  • Kim, Joo-Yeon;Olivry, Thierry;Son, Won-Geun
    • Journal of Veterinary Clinics
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    • v.29 no.2
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    • pp.141-147
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    • 2012
  • This work describes the characteristics of $Malassezia$ $pachydermatis$ isolated from dog ear canals and the effect of essential oils on the growth of this organism. Sterile cotton swabs were used to collect specimens from the external ear canal and culture tests were performed to detect the population size of $Malassezia$ yeast. Using three different isolation media, included Sabouraud dextrose agar (SDA) to isolate common $M.$ $pachydermatis$, and SDA supplemented with olive oil (SDAO) and Leeming's medium (LM) to detect lipophilic yeast, $Malassezia$ spp were isolated from 14 of 18 dogs (77.8%); isolation rates were 33.3% in SDA, 72.2% in SDAO and 66.7% in LM media. All $Malassezia$ spp isolates were identified as $M.$ $pachydermatis$ according to results of PCR amplification, but gross colony morphology and SDA growth rates suggested four different subtypes. Large (LC) and medium colony (MC) types respectively describe large colony (diameter > 3 mm) and medium colony (around 2 mm) after 72 hour incubation, and small (SC) type refers to smaller colony (< 1 mm) even after 5 days incubation; lipid dependent colonies did not grow onto SDA. Large Colony type strains were isolated from 4, 11, and 11 samples, MC type strains from 2, 3 and 1 and SC type strains from 1, 2 and 1 in SDA, SDAO and LM, respectively. Lipid-dependent $M.$ $pachydermatis$ (Lipo) were isolated from 3 samples each in SDAO and LM. Anti-$M.$ $pachydermatis$ activity testing was done using disc-diffusion assays and well diffusion tests. Most essential oils inhibited the growth of $M.$ $pachydermatis$ in a range from 0.5% to 1.0% of essential oils. MIC90 and MIC50 were variable depending upon the nature of essential oils. Thyme oil was found to be highly effective in inhibiting the growth of $M.$ $pachydermatis$ in a range from 0.125% to 0.0625% while marjoram and then tea tree oil exhibited lower inhibitory capacity.

Study on Affecting Variables Appearing through Chemical Pretreatments of Poplar Wood (Populus euramericana) to Enzymatic Hydrolysis (이태리 포플러의 화학적 전처리 공정을 통한 효소가수분해 영향 인자 분석)

  • Koo, Bon-Wook;Park, Nahyun;Yeo, Hwanmyeong;Kim, Hoon;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.3
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    • pp.255-264
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    • 2009
  • To evaluate the effects of chemical pretreatments of lignocellulosic biomass on enzymatic hydrolysis process, Populus euramericana was pretreated for 1 hr with 1% sulfuric acid ($H_2SO_4$) at $150^{\circ}C$ and 1% sodium hydroxide (NaOH) at $160^{\circ}C$, respectively. Before the enzymatic hydrolysis, each pretreated sample was subjected to drying process and thus finally divided into four subgroups; dried or non-dried acid pretreated samples and dried or non-dried alkali pretreated samples and chemical and physical properties of them were analyzed. Biomass degradation by acid pretreatment was determined to 6% higher compared to alkali pretreatment. By the action of acid ca. 24.5% of biomass was dissolved into solution, while alkali degraded ca. 18.6% of biomass. However, reverse results were observed in delignification rates, in which alkali pretreatment released 2% more lignin fragment from biomass to the solution than acid pretreatment. Unexpectedly, samples after both pretreatments were determined to somewhat higher crystallinity than untreated samples. This result may be explained by selective disrupture of amorphous region in cellulose during pretreatments, thus the cellulose crystallinity seems to be accumulated in the pretreated samples. SEM images revealed that pretreated samples showed relative rough and partly cracked surfaces due to the decomposition of components, but the image of acid pretreated samples which were dried was similar to that of the control. In pore size distribution, dried acid pretreated samples were similar to the control, while that in alkali pretreated samples was gradually increased as pore diameter increased. The pore volume which increased by acid pretreatment rapidly decreased by drying process. Alkali pretreatment was much more effective on enzymatic digestibility than acid pretreatment. The sample after alkali pretreatment was enzymatically hydrolyzed up to 45.8%, while only 26.9% of acid pretreated sample was digested at the same condition. The high digestibility of the sample was also influenced to the yields of monomeric sugars during enzymatic hydrolysis. In addition, drying process of pretreated samples affected detrimentally not only to digestibility but also to the yields of monomeric sugars.

Bioactive Utility of the Extracts from Rhus verniciflua Stokes (RVS) : Biological Function of the Extracts from RVS (옻나무 추출물의 생리활성 이용에 대한 연구 : 옻나무 추출물의 생물학적 기능)

  • Lim, Kye-Taek;Lee, Jeong-Chae
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.238-245
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    • 1999
  • Antioxidative effects of the water or ethanol extracts from Rhus verniciflua Stokes (RVS) were measured by protection against hydroxyl radicals in mouse brain tissue culture. In the water extracts from RVS, cell viabilities were estimated 60.0, 66.0, 72.0, 84.0 and 90.0% at addition of 1, 2, 4, 7 and $10{\mu}L$, respectively, compared with GO (20 mU/mL) alone. The cell viability in the ethanol extracts was similarly with water extracts. In the antitumor effects, the results showed that percentages of the HeLa cell death were approximately 24% for 12 hrs, 57% for 48 hrs at addition of 10%/well ethanol extracts respectively. To know inhibition of tumor growth, in vivo, mice (BALB/c) were inoculated with 0.25 mL CT-26 $(1{\times}10^6\;cells/mL)$ subcutaneously. After the generation of tumor, the results of RVS extracts (ethanol, water) injection showed generally that the tumor size in BALB/c was reduced. For physicochemical characterization of the RVS extracts, purified substances of water or ethanol extracts were analized with SDS-PAGE and ICP spectrometer. In electrophoresis, gel showed 2 bands (210, 230 KDa). The results of ICP verified that RVS extracts contain $Cu^{2+}$ in both samples. Conclusively, this substance might be a laccase which has a biological effective function, as a natural bioactive substance.

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Comparison of Gangjihwan and Combination of Gangjihwan and Gamisochehwan in the Improvement Effects of Weight Loss in a High Fat Diet-Fed Obese Mice (고지방식이 비만마우스 모델에서 파키스탄산 마황으로 조성된 강지환(降脂丸)의 단독 투여와 강지환합가미소체환(降脂丸合加味消滯丸)의 병용 투여의 체중감량 효과 비교)

  • Seok, Hoa Jun;Yoo, Jae Sang;Ku, Ja Ryong;Yoon, Ki Hyeon;Jo, Ju Heum;Jang, Du Hyon;Jung, Yang Sam;Kim, Jong Hoon;Kim, Byeong Chul;Roh, Jong Seong;Lee, Hye Rim;Lee, Hyunghee;Yoon, Michung;Shin, Soon Shik
    • Herbal Formula Science
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    • v.22 no.2
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    • pp.105-120
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    • 2014
  • Objectives : This study was investigated the improvement effects of Gangjihwan (DF) and combination of Gangjihwan and Gamisochehwan (GSH) on obesity in a high fat diet-fed obese mouse model. Methods : Eight-week-old C57BL/6N mice were divided into four groups: a normal lean group given a standard diet, an obese control group given a high fat diet, and DF and DF+GSH groups given a high fat diet with DF (40 mg/kg), and DF+GSH (80 mg/kg), respectively. After 8 weeks of treatment, body weight gain, feeding efficiency ratio, blood lipid markers, fat weight and histology were examined. Results : 1. Body weight gain and fat mass were significantly decreased in DF and DF+GSH groups compared with control. The extent of decreases was eminent in DF+GSH group. 2. Feeding efficiency ratio and circulating concentration of leptin were decreased in DF and DF+GSH groups compared with control. These decreases were significant in DF+GSH group. 3. Consistent with their effects on body weight gain and fat mass, circulating concentrations of triglyceride, glucose and insulin were decreased in DF and DF+GSH groups compared with control. 4. The size of adipocytes was decreased by DF and DF+GSH compared with control, whereas the adipocyte number per unit area was increased by them, suggesting that DF and DF+GSH decreased the number of large adipocytes. 5. Consistent with their effects on body weight gain, liver fibrosis was also improved in DF and DF+GSH groups compared with control. Conclusions : In conclusion, these results suggest that DF and DF+GSH groups decrease feeding efficiency ratio, plasma leptin concentration, blood anti-obesity biomarkers and fat mass, improves body weight gain contributing to the inhibition of liver fibrosis. In addition, these effects were more effective in DF+GSH combination group than in DF-only group.

A Study on Environmental Traits and Significance of Eouidongji Pond(於義洞池) Area in Dongchon(東村), Hanseong-bu(漢城府) (한성부 동촌(東村) 어의동지(於義洞池) 일대의 환경 특성과 장소적 의미)

  • Gil, Ji-Hye;Son, Yong-Hoon;Hwang, Kee-Won
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.35 no.2
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    • pp.1-14
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    • 2017
  • The purpose of this study is to understand the environmental traits and significance of the pond place that appears in historic maps in 'Dongchon(東村)', which is the eastern section within the fortress wall of Hanyang. We reviewed various documents and maps to identify the name of the pond, and of those materials we followed the name provided by official national records and geographical titles to designate it 'Eouidongji Pond(於義洞池).' The results of the research showed, first, that from the standpoint of location and environmental characteristics, that the area of the Eouidongji Pond was an area that contained a pastoral atmosphere within the city. It was located in an area where the waters flowing from the mountains gathered, and the boundaries of the pond would change based on water levels within a plot of land about 2,000pyeong(app. $6,600m^2$) in size. Second, when seen from the perspective of its creation background, the Eouidongji Pond seems to have been a naturally occurring pond which was modified when the water system was managed in the city at the beginning of the dynasty. In addition to its role as a reservoir, it was operated as a lotus pond(蓮池) to offer lotus(蓮) related by-products. With the nearby detached palace being actively used, it seems the pond was managed at the same time. The pond had already been filled in by the early 20th century, and although there were efforts to reconstruct the pond, it was eventually destroyed as the area was included in the site of a school during the colonial period. Third, the Eouidongji Pond was appreciated in many cases by individuals or admired as part of the natural landscape by viewing it from afar. In addition, it provided entry landscape near the road entering Dongchon, and was a location that was easily visited in connection with other nearby pleasure grounds. Through studying the location and environmental characteristics, the background of its creation and destruction, operation and management by time period, usage at the time, and characteristics of scenery appreciation of Eouidongji Pond, the Eouidongji Pond was a pond of high practical value to the nation, as it supplemented the water system in the city and was able to provide lotus harvests for the nation. In addition, from a urban environment perspective, it was a lateral landmark with a large area, as well as an effective boundary. It was an open area that the people of the Hanseong-bu could freely use, and it had a high public value due to its ease of accessibility.