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Inhibitory Effects of Ethanol Extracts from Pine Buds (Pinus densiflora) on Angiotensin Converting Enzyme, Xanthine Oxidase and Nitric Oxide Synthesis (소나무 새순 에탄올 추출물의 angiotensin converting enzyme, xanthine oxidase 및 nitrix oxide synthase 활성)

  • Cho, Eun-Kyung;Song, Hyo-Ju;Cho, Hea-Eun;Kim, Mi-Hyang;Choi, In-Soon;Choi, Young-Ju
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1629-1636
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    • 2009
  • Pine trees (Pinus densiflora Sieb. et Zacc.) have been used as a traditional health-promoting medicinal food in Korea. This research was performed to determine the antioxidative and antibacterial activities, tyrosinase, nitric oxide synthesis, angiotensin converting enzyme (ACE), and xanthine oxidase inhibition effects of the pine bud ethanol extract (PBE). Antioxidative activities of PBE were measured by using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) free radical scavenging activity and superoxide dismutase-like activity (SODA). DPPH radical scavenging and SOD-like activities of PBE were remarkably increased in a dose-dependent manner, and were about 88.9% and 47.9% at 1 mg/ml and 10 mg/ml, respectively. The xanthine oxidase and angiotensin converting enzyme activities were inhibited about 71.9% and 60.8% at 1 mg/ml and $100{\mu}g/ml$ of PBE, respectively. The tyrosinase inhibitory activities of PBE were slightly increased in a dose-dependent manner. The PBE showed strong antimicrobial activities on Escherichia coli (E. coli) and Vibrio paraheamolyticus. Stimulation of the macrophages RAW264.7 cells with lipopolysaccharide (LPS) resulted in increased production of nitric oxide (NO) in the medium. However, NO synthesis was reduced up to 54% by addition of PBE at $200{\mu}g/ml$. These results revealed that pine buds have a strong antioxidative and anti-inflammatory activity, and exhibit angiotensin converting enzyme and xanthine oxidase inhibitory activities. This suggests that pine buds have the greatest property as a source for natural health products.

Radiation Effect on NO, NOS and TGF-$\beta$ Expressions In Rat Lung (쥐의 폐에서 방사선이 Nitric Oxide (NO), Nitric Oxide Synthase (NO) 및 TGF- $\beta$의 발현에 미치는 영향)

  • Oh Young-Taek;Park Kwang-Joo;Kil Hoon-Jong;Ha Mahn Joon;Chun Mison;Kang Seung-Hee;Park Seong-Eun;Chang Sei-Kyung
    • Radiation Oncology Journal
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    • v.18 no.4
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    • pp.321-328
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    • 2000
  • Purpose :NOS2 induce NO Production and NO activate TGF-${\beta}$. The TGF-${\beta}$ is a inhibitor of NOS2. If this negative feedback mechanism operating in radiation pneumonitis model, NOS2 inhibitor may play a role in TGF-${\beta}$ suppression. We planned this study to evaluate the expression patterns of NO, NOS2 and TGF-${\beta}$ in vivo radiation pneumonitis model. Materials and Methods : Sixty sprague-Dawley rat were irradiated 5 Gy or 20 Gy. They were sacrificed 3, 7, 14, 28 and 56 days after irradiation. During sacrifice, we peformed broncho-alveolar lavage (BAL). The BAL fluids were centrifuged and supernatents were used for measure NO and TGF-${\beta}$, and the cells were used for RT-PCR. Results : After 5 Gy of radiation, NO in BAL fluid increased at 28 days in both lung and TGF-${\beta}$ in left lung at 56 days. NO increased in BAL fluid at 28 days in both lung after irradiation and TGF-${\beta}$ in right lung at 28-56 days after 20 Gy of radiation. After 5 Gy of radiation, NOS2 expression was increased in right lung at 14 days, in both lung at 28 days and in left lung at 56 days. TGF-${\beta}$ expression was reduced in both lung at 28 days and increased in left lung at 56 days. Conclusions :The Proposed feedback mechanism of NO, NOS2 and TGF-${\beta}$ was operated in vivo radiation pneumonitis model. At 56 days, however, NOS2 and TGF-${\beta}$ expressed concurrently in left lung after 5 Gy and in both lung after 20 Gy of radiation.

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Environmental Leachability of Electric Arc Furnace Dust for Applying as Hazardous Material Treatment (제강분진을 이용한 유해물질 처리기술 적용을 위한 안전성 평가)

  • Lee, Sang-Hoon;Kang, Sung-Ho;Kim, Jee-Hoon;Chang, Yoon-Seok
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.3
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    • pp.329-336
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    • 2006
  • Iron manufacturing process involves production of various by-product including slag, sludge, sintering and EAF(Electric Arc furnace dust). Some of the by-products such as EAF and sintering dust are disposed of as waste due to their high heavy metal contents. It has been notice for many years that the EAF dust also contain about 65% of Fe(0) and Fe(II) and then the possible utilization of the iron. One possibility is to apply the EAF as a lining material in conjunction with clay or HDPE liners, in waste landfill. The probable reaction between the leachate containing toxic elements such as TCE, PCE dioxine and $Cr^{6+}$ is reduction of the toxic materials in corresponding to the oxidation of the reduced iron and therefore diminishing the toxicity of the leachate. It is, however, prerequisite to evaluate the leaching characteristics of the EAF dust before application. Amelioration of the leachate would be archived only when the level of toxic elements in the treated leachate is less than that of in the untreated leachate. Several leaching techniques were selected to cover different conditions and variable environments including time, pH and contact method. The testing methods include availability test, pH-stat test and continuous column test. Cr and Zn are potentially leachable elements among the trace metals. The pH of the EAF dust is highly alkaline, recording around 12 and Zn is unlikely to be leached under the condition. On the contrary Cr is more leachable under alkaline environment. However, the released Cr should be reduced to $Cr^{3+}$ and then removed as $Cr(OH)_3$. Removal of the Cr is observed in the column test and further study on the specific reaction of Cr and EAF dust is underway.

STUDIES ON THE EXTRACTION OF SEAWEED PROTEINS 1. Extraction of Water Soluble Proteins (해조단백질의 추출에 관한 연구 1. 수용성 단백질의 추출)

  • RYU Hong-Soo;LEE Kang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.10 no.3
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    • pp.151-162
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    • 1977
  • Distribution of marine algae is diverse in Korea and the resource of edible algae is abundant marking 239,037 tons of yearly production in 1976. They have been known as a protein source and used as a supplement in Korean diet. It is necessary to estimate the potentiality and properties of usable algal proteins especially as food resources and studies of extraction and separation of the proteins, therefore, are basically required for this purpose. In this study, the influence of various factors including the sample treatment, extraction time and temperature, sample us extraction solvent ratio and pH upon the extractability of the water soluble protein was determined. And the effect of precipitation treatment for isolation of the algal protein from the extracts was also tested. Nine species of algae, the major ones in consumption as food namely Porphyra suborbiculata, Undaria pinnatifida, Hizikia fusiforme, Sargassum fulvellu, Enteromorpha linza, Codium fragile, Sargassum kjellmanianum and Ulva pertusa were collected as fresh from Kijang, Yangsan Gun, in the vicinity of Busan city. The content of crude protein $(N\times6.25)$ of the algae ranged from $9.46\%\;to\;24.14\% showing the highest value in Porphyra suborbiculata and the minimum in Hizikia fusiforme. In the effort of maceration of blending methods on the extractability, immersion freezing in dry ice-methanol solution appeared most effective yielding 1.5 to 2.5 times extractability than that of the mortar grinding method. The effect of the ratio of sample vs solvent on extractability differed from species. It was enhanced at the ratio of 1:20 (w/v) in Ulva pertusa and Enteromorpha linza while the ratio was 1:30 (w/v) for Cedium fragile, Undaria pinnatifida, Hizikia fusiferme, Sargassum fulvellum and Porphyra suborbiculata and 1:40 for Sargassum kjellmanianum respectively. The effect of extraction time and temperature was revealed differently from species which might be caused by differences in the constitution of algal tissues resulting in that the extraction for 1 hour at $50^{\circ}C$ gave the maximum extractabilily in Ulva pertusa and Enteromorpha linza, 2 hours in Porphyra suborbiculata, Hikikia fusiforme, Undaria pinnatifida, Sargassum kjellmanianum and 3 hours in Codium fragile. And the extractability was higher at $50^{\circ}C$ to $60^{\circ}C$ for the most of the tested samples except Hizikia fusiforme. The optimum pH for the extraction was 9 to 12. The recovery of extractable nitrogen to the total nitrogen was $63\%$ in average with the first extracts and $8.6\%$ with the second extracts respectively. Both extracts were prepared by 2 hour extraction at $50{\pm}1^{\circ}C$ with dry ice-methanol frozen and seasand macerated materials. And these conditions assumed to be an optimum for the extraction of water soluble algal proteins since the nitrogen content after the first extraction covered $90\%$ of the total water extractable nitrogen. In the precipitation of the extracted proteins, Barnstein method and methanol treatment seemed to be more efficient than other precipitation methods.

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Synthetic Chenodeoxycholic Acid Derivative HS-1200-Induced Apoptosis of Human Oral Squamous Carcinoma Cells (합성 Chenodeoxycholic Acid 유도체 HS-1200이 유도한 사람구강 편평상피암종세포 세포자멸사 연구)

  • Kim, In-Ryoung;Sohn, Hyeon-Jin;Kim, Gyoo-Cheon;Kwak, Hyun-Ho;Park, Bong-Soo;Choi, Won-Chul;Ko, Myung-Yun;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.32 no.3
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    • pp.251-261
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    • 2007
  • Bile acids and synthetic its derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. Previous studies have been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis inducing activity on various cancer cells in vitro. It wasn't discovered those materials have apoptosis induced effects on YD9 human oral squamous carcinoma cells. The present study was done to examine the synthetic bile acid derivatives(HS-1199, HS-1200) induced apoptosis on YD9 cells and such these apoptosis events. We administered them in culture to YD9 cells. Tested YD9 cells showed several lines of apoptotic manifestation such as activation of caspase-3, degradation of DFF, production of poly (ADP-ribose) polymerase(PARP) cleavage(HS-1200 only), DNA degradation(HS-1200 only), nuclear condensation, inhibition of proteasome activity, reduction of mitochondrial membrane potential(HS-1200 only) and the release of cytochrome c and AIF to cytosol. Between two synthetic CDCA derivatives, HS-1200 showed stronger apoptosis-inducing effect than HS-1199. Therefore HS-1200 was demonstrated to have the most efficient antitumor effect. Taken collectively, we demonstrated that a synthetic CDCA derivative HS-1200 induced caspases-dependent apoptosis via mitochondrial pathway in human oral sqauamous carcinoma cells in vitro. Our data therefore provide the possibility that HS-1200 could be considered as a novel therapeutic strategy for human orall squamous carcinoma from its poweful apoptosis-inducing activity.

Understanding Biotechnology: An Analysis of High School Students' Concepts (생명공학의 기본 개념에 대한 고등학생의 이해도 조사 및 개념 분석)

  • Chung, Young-Lan;Kye, Bo-Ah
    • Journal of The Korean Association For Science Education
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    • v.18 no.4
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    • pp.463-472
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    • 1998
  • Biotechnology is the process of using biological system for the production of materials. Genetic engineering, a subset of biotechnology, is the process of altering biological systems by the purposeful manipulation of DNA It is a new field in biology and no topic in biology is more likely to impact our personal lives and is therefore more worthy of our attention and understanding. The purpose of this study was to investigate students' understanding of the concepts of biotechnology, and a test tool which is made up of 20 basic questions was developed for the study. The subject of this study was high school students and the sample size was 486. In order to find out the source of students' misunderstanding, we also analysed high school textbooks and teachers were given the same tool applied to students. Two-way ANOVA was used for the analysis. Major findings of this study are as following; 1. Mean score of students was 41, and there was a significant difference between the scores of boys and girls(p<0.05). Female students scored higher than male students. The variables "region" and "major" had no significant influence. 2. Students' the most misunderstood concepts were "monoclonal antibody" and "gene cloning". Many students thought that a plamid DNA originally has a useful DNA in it, which is apparently wrong. 3. Mean score of teachers was 82, and the variabes of gender and career did not have statistically significant influence on the result(p>0.05). 4. Teachers got the lowest scores on the concepts of "gene therapy", "the accomplishment of biotechnology in agriculture and medicine", and "plasmid DNA". The results of item analysis implied that teachers' misunderstanding might be a part of the sources of students' misunderstaning. 5. Out of 18 basic concepts selected in the study, only 10 concepts were explained well enough in most textbooks. The results of item analysis indicated that textbooks also could be a part of the source of students' misunderstanding.

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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.

Gel and Texture Properties of Fish-meat Gel Prepared with Pagrus major in Comparison to Different Grades of Alaska Pollock (도미를 활용하여 제조한 연제품의 겔 및 texture 특성)

  • Gao, Ya;Oh, Jung Hwan;Karadeniz, Fatih;Lee, Seul-Gi;Kim, Hyung Kwang;Kim, Se Jong;Jung, Jun Mo;Cheon, Ji Hyeon;Kong, Chang-Suk
    • Journal of Life Science
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    • v.26 no.8
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    • pp.955-962
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    • 2016
  • Fish-meat gel is an intermediate product used in a variety of surimi-based seafood. One of the most-used raw materials of fish-meat gel is Alaska Pollock due to its high-quality meat in terms of gel strength and texture. However, increasing demand for fish-meat gel, along with overexploitation of the wild catch Alaska Pollock, has put the industry in need of low-cost sustainable alternative sources for fish-meat gel. Pagrus major (PM) is a widely aquacultured fish known for having white meat that is low in fat. The current study compares the quality of fish-meat gel prepared from aquacultured PM to that of high and mid-grade Alaska Pollock fish-meat gel. Gels were compared in terms of gel strength, texture, color, and protein pattern. Results indicated that fish-meat gels prepared from PM were superior to Alaska Pollock fish-meat gels with regard to gel strength, hardness, springiness, chewiness, cutting strength, and breaking force. In addition, although not matching in quality, PM exhibited a cohesiveness, whiteness, and expressible moisture content comparable to Alaska Pollock of both grades. Protein pattern analysis also showed that PM and Alaska Pollock fish-meat gels had similar protein profiles before and after gel preparation. Therefore, P. major is suggested as a potential substitute for Alaska Pollock in fish-meat gel production.

Skin-Whitening and UV-Protective Effects of Angelica gigas Nakai Extracts on Ultra High Pressure Extraction Process (초고압 추출 공정에 의한 당귀 추출물의 미백 및 자외선 차단 효과)

  • Kim, Cheol-Hee;Kwon, Min-Chul;Han, Jae-Gun;Na, Chun-Su;Kwak, Hyeong-Geun;Choi, Geun-Pyo;Park, Uk-Yeon;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.4
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    • pp.255-260
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    • 2008
  • This study was performed to investigate the enhancement of UV-protection activities and skin-whitening effects from Angelica gigas Nakai extracts on ultra high pressure extraction process. Extraction at $60^{\circ}C$ treated by ultra high pressure for 15 minute and associated with ultrasofication (HPE15) was showed more than double yield, compare conventional extraction, as 12.24% (w/w) from A. gigas. Extracts of HPE15 reduced expression of MMP-1 on UV-irradiated CCD-986sk cells as 122.2% and revealed high inhibitory potency on tyrosinase as 69.4% by adding samples. Extracts of HPE15 from A. gigas showed strong inhibition effect on melanin production test by Clone M-3 cells as 82.4% by adding extracts. From the preliminary observations, we considered that the extracts from A. gigas could be potent natural materials for skin-whitening agent, and could be used as a potential anti-aging agent for the photo-damaged skin.

Inhibitory Effects of Ethanolic Extracts from Aster glehni on Xanthine Oxidase and Content Determination of Bioactive Components Using HPLC-UV (섬쑥부쟁이 에탄올 추출물의 잔틴산화효소 저해 효능 및 HPLC-UV를 이용한 유효성분의 함량 분석)

  • Kang, Dong Hyeon;Han, Eun Hye;Jin, Changbae;Kim, Hyoung Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.11
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    • pp.1610-1616
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    • 2016
  • This study aimed to establish an optimal extraction process and high performance liquid chromatography-ultraviolet (HPLC-UV) analytical method for determination of 3,5-dicaffeoylquinic acid (3,5-DCQA) as a part of materials standardization for the development of a xanthine oxidase inhibitor as a health functional food. The quantitative determination method of 3,5-DCQA as a marker compound was optimized by HPLC analysis using a Luna RP-18 column, and the correlation coefficient for the calibration curve showed good linearity of more than 0.9999 using a gradient eluent of water (1% acetic acid) and methanol as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 320 nm. The HPLC-UV method was applied successfully to quantification of the marker compound (3,5-DCQA) in Aster glehni extracts after validation of the method with linearity, accuracy, and precision. Ethanolic extracts of A. glehni (AGEs) were evaluated by reflux extraction at 70 and $80^{\circ}C$ with 30, 50, 70, and 80% ethanol for 3, 4, 5, and 6 h, respectively. Among AGEs, 70% AGE at $70^{\circ}C$ showed the highest content of 3,5-DCQA of $52.59{\pm}3.45mg/100g$ A. glehni. Furthermore, AGEs were analyzed for their inhibitory activities on uric acid production by the xanthine/xanthine oxidase system. The 70% AGE at $70^{\circ}C$ showed the most potent inhibitory activity with $IC_{50}$ values of $77.01{\pm}3.13{\sim}89.96{\pm}3.08{\mu}g/mL$. The results suggest that standardization of 3,5-DCQA in AGEs using HPLC-UV analysis would be an acceptable method for the development of health functional foods.