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EFFECT OF NERVE GROWTH FACTOR GENE INJECTION ON THE NERVE REGENERATION IN RAT LINGUAL NERVE CRUSH-INJURY MODEL (백서 설신경 압박손상모델에서 신경성장인자 유전자 주입이 신경재생에 미치는 영향)

  • Gao, En-Feng;Chung, Hun-Jong;Ahn, Kang-Min;Kim, Soung-Min;Kim, Yun-Hee;Jahng, Jeong-Won;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.5
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    • pp.375-395
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    • 2006
  • Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

Scheduling Algorithms and Queueing Response Time Analysis of the UNIX Operating System (UNIX 운영체제에서의 스케줄링 법칙과 큐잉응답 시간 분석)

  • Im, Jong-Seol
    • The Transactions of the Korea Information Processing Society
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    • v.1 no.3
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    • pp.367-379
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    • 1994
  • This paper describes scheduling algorithms of the UNIX operating system and shows an analytical approach to approximate the average conditional response time for a process in the UNIX operating system. The average conditional response time is the average time between the submittal of a process requiring a certain amount of the CPU time and the completion of the process. The process scheduling algorithms in thr UNIX system are based on the priority service disciplines. That is, the behavior of a process is governed by the UNIX process schuduling algorithms that (ⅰ) the time-shared computer usage is obtained by allotting each request a quantum until it completes its required CPU time, (ⅱ) the nonpreemptive switching in system mode and the preemptive switching in user mode are applied to determine the quantum, (ⅲ) the first-come-first-serve discipline is applied within the same priority level, and (ⅳ) after completing an allotted quantum the process is placed at the end of either the runnable queue corresponding to its priority or the disk queue where it sleeps. These process scheduling algorithms create the round-robin effect in user mode. Using the round-robin effect and the preemptive switching, we approximate a process delay in user mode. Using the nonpreemptive switching, we approximate a process delay in system mode. We also consider a process delay due to the disk input and output operations. The average conditional response time is then obtained by approximating the total process delay. The results show an excellent response time for the processes requiring system time at the expense of the processes requiring user time.

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The Change of Antioxidant Enzyme (Superoxide Dismutase, Catalase, Glutathione Peroxidase) in the Endotoxin Infused Rat Lung (내독소 투여후 쥐의 폐조직내 Antioxidant (Superoxide Dismutase, Catalase, GSH-Peroxidase)의 변화에 대한 연구)

  • Song, Jeong-Sup;Kim, Chi-Hong;Kwon, Soon-Seog;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Han, Ki-Don;Moon, Hwa-Sik;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.2
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    • pp.104-111
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    • 1993
  • Background: Gram-negative bacterial endotoxin induced septicemia is known to be a leading cause in the development of adult respiratory distress syndrome(ARDS). The mechanism of endotoxin induced lung injury is mainly due to the activated neutrophils which injure the capillary endothelial cells by releasing oxidant radical and resulted in pulmonary edema. We studied the change of antioxidant enzyme in the case of large or small, intermittant dose of endotoxin infused rat lungs. Methods: Endotoxin was given to the rat through the peritoneal cavity in the dose of 7 mg/kg body weight in the large dose group and 1 mg/kg for 10 days in the small dose group. Bronchoalveolar lavage (BAL) was done and rats were killed at 6, 12, 24 hours after single endotoxin injection in the large dose group and 3, 7, 10 days after daily endotoxin injection for 10 days in the small dose group. The lungs were perfused with normal saline through the pulmonary artery to remove the blood and were homogenized in 5 volume of 50 mM potassium phosphate buffer containing 0.1 mM EDTA. After centrifuging at 100,000 g for 60 minute, the supernatent was removed and stored at $-70^{\circ}C$ until measuring for superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px) and protein. Results: We observed the following results. 1) The lung wet/dry weight ratio and albumin concentration in the BAL fluids were increased to peak at 12 hours and neutrophil number in the BAL fluids were peak at 6 hours after endotoxin injection in the large dose group. 2) Cu, Zn SOD (IU/mg protein) was significantly decreased after 6, 12 hours after endotoxin injection in the large dose group. 3) There were no singnificant change in the level of Mn SOD, catalase, GSH-Px after endotoxin injection in both groups. Conclusion: Endotoxin in the large dose group produced the acute pulmonary edema and decreased the Cu, Zn SOD in the lung tissue after injecting endotoxin at 6 and 12 hours. These phenomenon may be due to the cell membrane damage by endotoxin. Further research would be necessary whther giving SOD by intratracheal route or method to increase the synthesis of SOD may lessen the acute lung injury by endotoxin.

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Enhanced Growth Inhibition by Combined Gene Transfer of p53 and $p16^{INK4a}$ in Adenoviral Vectors to Lung Cancer Cell Lines (폐암세포주에 대한 p53 및 $p16^{INK4a}$의 복합종양억제유전자요법의 효과)

  • Choi, Seung -Ho;Park, Kyung-Ho;Seol, Ja-Young;Yoo, Chul-Gyu;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.67-75
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    • 2001
  • Background : Two tumor suppressor genes, p53 and p16, which have different roles in controlling the cell cycle and inducing apoptosis, are frequently inactivated during carcinogenesis including lung cancer. Single tumor suppressor gene therapies using either with p53 or p16 have been studied extensively. However, there is a paucity of reports regarding a combined gene therapy using these two genes. Methods : The combined effect of p53 and p16 gene transfer by the adenoviral vector on the growth of lung cancer cell lines and its interactive mechanism was investigated. Results : An isobologram showed that the co-transduction of p53 and p16 exhibited a synergistic growth in hibitory effect on NCI H358 and an additive effect on NCI H23. Cell cycle analysis demonstrated the induction of a synergistic G1/S arrest by a combined p53 and p16 transfer. This synergistic interaction was again confirmed in a soft agar confirmed in a soft agar clonogenic assay. Conclusion : These observations suggest the potential of a p53 and p16 combination gene therapy as another potent strategy in cancer gene therapy.

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Application of Noninvasive Positive Pressure Ventilation in Patients with Respiratory Failure (호흡부전 환자에서 비침습적 양압환기법의 적용)

  • Seol, Young Mi;Park, Young Eun;Kim, Seo Rin;Lee, Jae Hyung;Lee, Su Jin;Kim, Ki Uk;Cho, Jin Hoon;Park, Hye Kyung;Kim, Yun Seong;Lee, Min Ki;Park, Soon Kew;Kim, Young Dae
    • Tuberculosis and Respiratory Diseases
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    • v.61 no.1
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    • pp.26-33
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    • 2006
  • Background: Noninvasive positive pressure ventilation(NPPV) has been increasingly used over the past decade in the management of acute or chronic respiratory failure and weaning of mechanical ventilation. We performed this clinical study to evaluate the usefulness of NPPV in patients who developed acute respiratory failure or post-extubation respiratory failure. Methods: We analysed thirty four patients(sixteen males and eighteen females, mean ages 58 years) who applied NPPV(BIPAP S/T, Respironics co., USA) for respiratory failure or weaning difficulty at medical intensive care unit(MICU), emergency room and general ward of a tertiary hospital. We evaluated the underlying causes of respiratory failure, duration of treatment, the degree of adaptation, complication and predictive parameters of successful outcome. Results: The overall success rate of NPPV was seventy-one percent. The duration of NPPV applying time, baseline blood pressure, pulse rate, respiration rate, $PaO_2$, $PaCO_2$, $SaO_2$ were not different between success group and failure group. But, the baseline pH was higher in the success group. Predictors of success were higher baseline pH, patients with underlying disease of COPD, improvement of vital sign and arterial blood gas value after NPPV application. The success rate in patients with post-extubation respiratory failure was eighty percent. There were no serious complication on applying NPPV except minor complications such as facial skin erythema, abdominal distension & dry mouth. Conclusion: NPPV may be effective treatment in patients with acute respiratory failure or post-extubation respiratory failure in selected cases.

Biological stability of Zirconia/Alumina composite ceramic Implant abutment (지르코니아/알루미나 복합 지대주의 생물학적 안정성에 관한 연구)

  • Bae, Kyu-Hyun;Han, Jung-Suk.;Kim, Tae-Il;Seol, Yang-Jo;Lee, Yong-Moo;Ku, Young;Cho, Ki-Young;Chung, Chong-Pyoung;Han, Soo-Boo;Rhyu, In-Chul
    • Journal of Periodontal and Implant Science
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    • v.36 no.2
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    • pp.555-565
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    • 2006
  • The purpose of the present study is to evaluate the biological stability of the zirconia/alumina composite abutment by histologic and radiographic examination in clinical cases. 17 partially edentulous patients (5 men and 12 women, mean age 47) were treated with 37 implants. The implants were placed following the standard two-stage protocol. After a healing period of 3 to 6 months, zirconia/alumina composite abutments were connected. All radiographs were taken using paralleling technique with individually fabricated impression bite block, following insertion of the prosthesis and at the 3-, 6-, 12 month re-examinations. After processing the obtained images, the osseous level was calculated using the digital image in the mesial and distal aspect in each implant. An ANOVA and t-test were used to test for difference between the baseline and 3-, 6-, 12 months re-examinations, and for difference between maxilla and mandible. Differences at P <0.05 were considered statistically significant. For histologic examination, sample was obtained from the palatal gingiva which implant functioned for 12 months. Sections were examined under a light microscope under various magnifications. Clinically, no abutment fracture or crack as well as periimplantitis was observed during the period of study. The mean bone level reduction(${\pm}standard$ deviation) was 0.34 rom(${\pm}\;0.26$) at 3-months, 0.4 2mm(${\pm}\;0.30$) at 6-months, 0.62 mm(${\pm}\;0.28$) at 12-months respectively. No statistically significant difference was found between baseline and 3-, 6-, 12-months re-examinations (p > 0.05). The mean bone level reduction in maxilla was 0.33(${\pm}0.25$) at 3-months, 0.36(${\pm}0.33$) at 6-months, 0.56(${\pm}0.26$) at 12-months. And the mean bone level reduction in mandible was 0.35(${\pm}0.27$) at 3-months, 0,49(${\pm}0.27$) at 6-months, 0.68(${\pm}0.30$) at 12-months. No statistical difference in bone level reduction between implants placed in the maxilla and mandible. Histologically, the height of the junctional epithelium was about 2.09 mm. And the width was about 0.51 mm. Scattered fibroblasts and inflammatory cells, and dense collagen network with few vascular structures characterized the portion of connective tissue. The inflammatory cell infiltration was observed just beneath the apical end of junctional epithelium and the area of direct in contact with zirconia/alumina abutment. These results suggest the zirconia/alumina composite abutment can be used in variable intraoral condition, in posterior segment as well as anterior segment without adverse effects.

The effects of enamel matrix derivatives on the proliferation and gene expression of PDL fibroblast, $SaOs_2$ cells and Cementum derived cells (법랑기질유도체가 치주인대세포, 불멸화 조골세포, 백악질 유래세포의 증식과 유전자 발현에 미치는 영향)

  • Jeong, Yoo-Jee;Kim, Kyoung-Hwa;Kim, Tae-Il;Seol, Yang-Jo;Ku, Young;Lee, Hae-Jun;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Lee, Yong-Moo
    • Journal of Periodontal and Implant Science
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    • v.35 no.2
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    • pp.321-333
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    • 2005
  • 1. 목적 in vitro 상에서 법랑기질유도체가 치주인대섬유아세포, 불멸화 조골세포와 백악질 유래세포의 증식과 유전자 발현에 미치는 영향을 알아보고자 하였다. 2. 연구방법 및 재료 <세포증식 연구> 교정을 목적으로 발거한 치아에서 분리, 배양한 치주인대섬유아세포와 백악질유래세포, 그리고 $SaOs_2$ 세포를 이용하였다. 법랑기질유도체가 세포 증식에 미치는 영향을 알아보기 위해, 35 mm Petri dish에 dish 당 $5{\times}10^3$ 개의 세포를 접종하였다. 대조군은 1% 항생제와 10% FBS를 포함한 DMEM 배지를 이용했고, 5mM 초산을 첨가한 군과 첨가하지 않은 두 개의 대조군이 이용되었다. 실험군은 100 ${\mu}g/ml$의 법랑기질유도제를 첨가한 군과 100 ${\mu}g/ml$의 법랑기질유도체와 5 mM의 초산을 첨가한 2개의 실험군이 이용되었다. 각 군은 세 개의 배양접시에 행해졌고, 1, 3, 8일에 세포의 수를 각각 측정하였다. 결과는 repeated measures ANOVA로 통계 처리하였다. <유전자 발현 연구> 각 세포의 형질 특성을 알아보기 위해 RT-PCR을 실시하여 조골세포 분화 표식자와 연관된 Human collagen type I(COL I), human osteopontin(OP), human osteocalcin(OC), human alkaline phosphatase(ALP)와 human bone sialoprotein(BSP)의 mRNA 발현을 실험 1, 3, 8일에 걸쳐, 세 군의 차이를 비교 관찰하였다. 3. 결과 <세포증식 연구> 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포의 증식은 법랑기질유도체에 의해 영향을 받지 않았다. 대조군과 초산이 포함된 대조군 그리고 법랑기질유도체와 초산이 포함된 실험군에서 유의할 만한 세포 수의 차이가 실험 기간 1, 3, 8일에 걸쳐 나타나지 않았다(p<0.05). <유전자 발현 연구> ALP와 COL I은 세 군의 세포에서 모두 발현되었고, 발현 정도는 EMD에 영향을 받지 않았다. OC은 세 군에서 모두 비교적 약하게 발현되었고, 특히 $SaOs_2$ cell과 백악질유래세포에서 약하게 발현되었다. EMD는 OC의 발현정도를 약하게 하였다. OP은 백악질유래세포에서 1, 3, 8일에 걸쳐 EMD 유무에 관련 없이 발현되지 않았다. 그러나 치주인대세포와 $SaOs_2$ cell에서는 강하게 발현되었다. BSP는 치주인대세포와 $SaOs_2$ cell에서 1, 3, 8일에 걸쳐 비교적 고르게 발현되었다. EMD 배지에서 배양된 백악질유래세포는 8일에는 BSP가 발현되지 않았다. 4 결론 이번 실험 결과에 의하면 법랑기질유도체는 치주인대세포, 불멸화 조골세포와 백악질 유래세포의 증식에 있어 유의성 있는 효과를 나타내지 않았다. 그러나, 유전자 발현에 있어서는, 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포 모두에서 OC mRNA의 발현을 억제하는 효과를 나타내었다. EMD는 세포의 증식에는 영향을 미치지 않지만, 유전자 발현에 있어 일부 영향을 미치는 것으로 보인다. 법랑기질유도체가 세포의 증식과 유전자 발현에 미치는 영향은 배양된 세포의 형질특성, 배양환경, 배양일수 등에 따라 달라질 수 있다. 그러므로 법랑기질유도체가 in vitro 상에서 세포에 미치는 영향은 보다 정량화된 연구가 필요하다.

A Study on HanYongUn's Sijo (한용운 시조의 내면 세계와 표현 미학)

  • Jeon, Jae-Gang
    • Sijohaknonchong
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    • v.43
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    • pp.177-206
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    • 2015
  • This paper is written in order to research for the contents and expression of HanYongUn's Sijo. HanYongUn is very famous as monk and independent campaigner, modern poet in Korea. He wrote many kinds of literary works, for example, many modern poetry, modern novels, Sijo, Chino-Korean Poetry etc. It's very exceptional that he wrote a lot of Korean traditional Sijo and Chino-Korean Poetry. Because he was a many modern poet as same as modern novelist. So studying on his Sijo can help someone to understand the essence of HanYongUn's all literature. That's why I'm studying on HanYongUn's Sijo. The firstly, in aspect of the the contents of HanYongUn's Sijo, HanYongUn was expressing three kinds of themes, that is ideology, reality, daily life in his Sijo. The ideology consists of Buddhism and Confucianism and the reality is related with social conditions, the daily life is deeply connected with Nim. These features of his Sijo are different from his modern poetry and Chino-Korean Poetry which had a simple theme, for example, love with Nim, daily life. The secondly, in aspect of the expression of HanYongUn's Sijo, I studied the expression of HanYongUn's Sijo in three angles, that is, vocabulary and the developing of poet thinking, rhetorics. HanYongUn used essential words for expressing three kinds of themes effectively in his Sijo. And he was developing of his poet thinking by three steps in his Sijo. He applied several representative rhetorics to his Sijo, those are question and answer, exclamation, irony, distich etc. Even though I studied the characteristics of HanYongUn's Sijo in two aspects But there could be the other things to study about these kinds of theme. I might continue researching the other kinds of theme next time in the near future.

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Antioxidant Activity of Rubus crataegifolius Bge. Fruit Extracts (Rubus crataegifolius Bge. 열매 추출물의 항산화 활성)

  • Moon, Kyoung-Mi;Kim, Ji-Eun;Kim, Hae-Young;Lee, Jae-Seol;Son, Gi-Ae;Nam, Soo-Wan;Kim, Byung-Woo;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1214-1218
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    • 2011
  • We investigated the fruits of Rubus crataegifolius Bge, a plant which has been traditionally used in Korea in phytotherapy, to describe antioxidant materials from plant sources. R. crataegifolius fruits were extracted with methanol and further fractionated into n-hexane, diethyl ether, and ethyl acetate. The antioxidant activity of each fraction and the residue was assessed using a 1,1-diphenyl-2-picrylhydrazyl (DPPH), $H_2O_2$ radical scavenging method, and their cytotoxicity on human primary kerationcyte (HK) was determined by an MTS assay. The R. crataegifolius fruit methanol extract showed strong antioxidant activity (75.04%, 50%) compared with vitamin C (79.9%, 54.1%) by the DPPH, and $H_2O_2$ method, respectively. The measured activity from the subsequent extracts of the methanol extract were 20.3% for n-hexane fraction (HF), 68.8% for diethyl ether fraction (DF), 67.1% for ethyl acetate fraction (EF), and 67.1% for the residue fraction (RE) by DPPH and 2.2% for HF, 1.6% for DF, 10% for EF, and 50% for the RE by $H_2O_2$ assay. An oxidative stress model of HK was established under a suitable concentration (1 mM). The cell viability of the RE treated group increased and the percentage of apoptotic cells decreased at concentrations of 0.005-0.02% RE compared with the $H_2O_2$ treated group. Fruit extracts of the medicinal plant R. crataegifolius showed potent antioxidant activity and the ability to relieve cell damage from $H_2O_2$ induced injury to HK.

Study on Korean Ancient Dietary Culture through Japanese Sacrificial Offerings -Danjanjinja Kakitsisai(2)- (일본신찬(日本神饌)을 통한 한국고대식(韓國古代食)의 추정연구(推定硏究) -담산신사(談山神社) 가길제(嘉吉祭) 백미어식(百味御食)(2)-)

  • Kim, Chon-Ho;Kaneko, Kentaro;Sumino, Takeshi;Kaneda, Takashi
    • Journal of the Korean Society of Food Culture
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    • v.8 no.2
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    • pp.139-146
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    • 1993
  • The predeccessor of Danjanjinja was Myorak temple which is built in the 7th century. At that age, the Buddhist culture of Japan had highly prospected by transmitting Buddhism to Japan from Han peninsular On the other hand, the private god of Fujiwara family in Danjanjinja is Uchigami which is one of typical Japanese popular belief like Dangshin of Korean's. Through these historical background, it could by presumed that the Kakitsisai might be the original form of Korean Buddhist sacrificial offerings from ancient age. So this study on Kakitsisai what had handed down from generation to generation about for 1300 years help us to study and estimate the ancient dietary culture of Korean and Japanese. 1. Kakitsisai performed high filling method in the sacrificial offerings like Kasuga, Horyuji and Korea. 2. The patterns and colors of high filling offerings are various in Korea and Japan. 3. They used unpolished rice by ancient rice, and called red and black one. We can guess both of countries ate unpolished rice at that age. 4. They used many kind of ancient wild fruits and vegetables. We could recognize what the ancients had eaten the foods.

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