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Enzymatic Preparation and Antioxidant Activities of Protein Hydrolysates from Protaetia brevitarsis Larvae (흰점박이꽃무지 유충 단백가수분해물의 제조 및 항산화 활성)

  • Lee, Hyo-Seon;Ryu, Hee-Jeong;Song, Hyeon-Ji;Lee, Syng-Ook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.10
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    • pp.1164-1170
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    • 2017
  • Protaetia brevitarsis larvae (PBL) has recently been registered as a temporary food in Korea, and this study evaluated the application potential of PBL proteins as health functional food materials. Protein hydrolysates were prepared from PBL powder by enzymatic hydrolysis using five different proteases (alcalase, bromelain, flavourzyme, neutrase, and papain), and based on the results from the peptide content and SDS-PAGE analyses, PBL treated with alcalase or flavourzyme showed a high degree of hydrolysis (HD) value, whereas the HD value of those treated with neutrase, bromelain, or papain was minimal. The protein hydrolysates showing a high HD value were separated further into the fractions of >3 kDa and <3 kDa by a centrifugal filter system and then lyophilized, and according to the $RC_{50}$ values of the protein hydrolysates (<3 kDa) obtained from three different antioxidant analyses; the alcalase hydrolysates showed the highest antioxidant activity. Therefore, the alcalase hydrolysates were tested further for their inhibitory effects on the peroxidation of linoleic acid by measuring the thiobarbituric acid values. The results showed that the peroxidation of untreated linoleic acid increased dramatically during 6 days of incubation, but a pretreatment with the hydrolysates ($100{\sim}800{\mu}g/mL$) significantly inhibited the linoleic acid peroxidation in a dose-dependent manner for 6 days. Our current studies are focused on the identification of active peptide sequences from alcalase hydrolysates.

Effects of epigallocatechin gallate on $CoCl_2-induced$ apoptosis in PC12 cells (PC12 세포에서 $CoCl_2$ 유발 세포자멸사에 대한 epigallocatechin-gallate의 역할)

  • Mo, Hyun-Chul;Choi, Nam-Ki;Kim, Seon-Mi;Kim, Won-Jae;Yang, Kyu-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.1
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    • pp.13-24
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    • 2006
  • Neuronal apoptotic events, consequently resulting in neuronal cell death, are occurred in hypoxic/ischemic condition. This cell death has been shown to be accompanied with the production of reactive oxygen species (ROS), which can attack cellular components such as nucleic acids, proteins and phospholipid. However, the underlying mechanisms of apoptosis induced in hypoxic/ischemic condition and its treatment methods are unsettled. Cobalt chloride $(CoCl_2)$ has been known to mimic hypoxic condition including the production of ROS. Epigallocatechin gallate (EGCG), a green tea polyphenol, has diverse pharmacologial activities in cell growth and death. This study was aimed to investigate the apoptotic mechanism by $CoCL_2$ and effects of EGCG on $CoCl_2-induced$ apoptosis in PC12 cells. Administration of $CoCl_2$ decreased cell survival in dose- and time-dependent manners and induced genomic DNA fragmentation. Treatment with $100{\mu}M$ EGCG for 30 min before PC12 cells were exposed to $150{\mu}M$ $CoCl_2$, being resulted in the cell viability and DNA fragmentation being rescued. $CoCl_2$ caused morphologic changes such as cell swelling and condensed nuclei whereas EGCG attenuated morphologic changes by $CoCl_2$. EGCG suppressed the apoptotic peak and a loss of ${\Delta}{\psi}_m$ induced by $CoCl_2$. $CoCl_2$ decreased Bcl-2 expression but Bax expression was not changed in $CoCl_2$- treated cells. EGCG attenuated the Bcl-2 underexpression by $CoCl_2$. $CoCl_2$ augumented the cytochrome c release from mitochondria into cytoplasm and increased caspase-8, -9 and caspase-3 activity a marker of the apoptotic executing stage. EGCG ameliorated the incruement in caspase-8, -9 and -3 activity, and cytochrome c release by $CoCl_2$ NAC (N-acetyl-cysteine), a scavenger of ROS, attenuated $CoCl_2-induced$ apoptosis in consistent with those of EGCG. These results suggest that $CoCl_2$ induces apoptotic cell death through both mitochondria- and death receptor-dependent pathway and EGCG has neuroprotective effects against $CoCl_2-induced$ apoptosis in PC12 cells.

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Change of Frequency of Chromosome Aberration by Time Interval after Radiation Therapy (방사선 치료후 시간경과에 따른 염색체이상 빈도의 변화)

  • Kim, Mi-Sook;Yi, Chun-Ja;Ha, Sung-Whan;Song, Myung-Jae;Kim, Hee-Jeun
    • Journal of Radiation Protection and Research
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    • v.19 no.1
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    • pp.51-68
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    • 1994
  • It is good method to use frequency of chromosome aberration in Lymphocytes for a biological dosimetry in cases of accidental exposure to radiation. But in cases of past exposure, biological dosimetry is limited because the frequency of aberration decreases by time after exposure. To provide a basic data for estimation of past radiation exposure, the changing pattern of frequency of unstable chromosome aberration by time interval after exposure was studied. Observation was made on peripheral lymphocytes of 41 blood samples from 20 patients treated for uterine cervical carcinoma and endometrial carcinoma. The patients received 50.4Gy radiation to whole pelvis. Elapsed times after the completion of radiation therapy were 1 day, 3, 6, 9, 12, 24, 52, 104, 156, 208, 260 and 520 weeks. All the blood sample were microcultured. The Ydr, Qdr and Qdra were calculated from frequency of unstable aberration. Ydr did not decrease for 3 weeks after radiation therapy, and thereafter, decreased very rapidly and reached 0.05 at two years after radiation therapy and decreased very slowly until 5 years after radiation therapy. Relationship between unstable chromosome aberration and time interval after radiation therapy was described as $Ydr=0.259{\times}exp(-0.0429T)+0.0560{\times}exp(-0.00106T)$ (time in weeks) Qdr remained constant at 1.51 until 24 weeks after radiation therapy and then decreased to 1.17 at 52 weeks. Therafter, it did not change. Qdra remained constant at 1.10 for 12 weeks after radiation therapy and decreased to 0.81 at 52 weeks. Thereafter, it remained constant. Two superimposed exponential Ydr disappearance rate suggests that it is possible to calculate the past exposure dose. When the elapsed time after exposure is short, Qdr and Qdra are useful papameters for biological dosimetry for past radiation exposure.

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The Combined Therapy of Inhaled Nitric Oxide and Prone Positioning Has an Additive Effect on Gas Exchange and Oxygen Transport in Patients with Acute Respiratory Distress Syndrome (급성호흡곤란증후군 환자에서 복와위(prone position)와 산화질소흡입(nitric oxide inhalation) 병용 치료의 효과)

  • Koh, Youn-Suck;Lim, Chae-Man;Lee, Ki-Man;Chin, Jae-Yong;Shim, Tae-Sun;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1223-1235
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    • 1998
  • Background and Objective : Although prone positioning has been reported to improve gas exchange, prone positioning alone does not seem to be sufficient to increase systemic oxygen transport in an acute lung injury. The objective of this study was to investigate whether the combined therapy of low dose nitric oxide (NO) inhalation and prone positioning has an additive effect on the oxygenation and hemodynamics in patients with severe ARDS. Patients and Methods : Twelve patients with ARDS were included. Prone positioning alone, later combined with nitric oxide inhalation (5~10 ppm) from the supine position (baseline) were performed with serial measurement of gas exchange, respiratory mechanics and hemodynamic at sequential time points. The patient was regarded as a responder to prone positioning if an increase in $PaO_2/FiO_2$ of more than 20 mm Hg at 30 min or 120 min intervals after prone positioning was observed compared to that of the baseline. The same criterion was applied during nitric oxide inhalation. Results : Eight patients (66.5%) responded to prone positioning and ten patients (83.3%) including the eight just mentioned responded to the addition of NO inhalation. The $AaDO_2$ level also decreased promptly with the combination of prone positioning and NO inhalation compared to that of prone positioning alone ($191{\pm}109$ mm Hg vs. $256{\pm}137$ mm Hg, P<0.05). Hemodynamic parameters and lung compliance did not change significantly during prone positioning only. Following the addition of NO inhalation to prone positioning, the mean pulmonary artery pressure and pulmonary artery occlusion pressure decreased and cardiac output, stroke volume and oxygen delivery increased (P < 0.05) compared to those of prone 120 min. Conclusion : These findings indicate that NO inhalation would provide additional improvement in oxygenation and oxygen transport to mechanically ventilated patients with ARDS who are in a prone position.

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A Study of EFFECT and MECHANISM of IL-2 on SURVIVAL of EOSINOPHILS (Interleukin-2가 호산구 생존에 미치는 영향과 가전에 관한 연구)

  • Kim, Hyo-Seok;Lee, Young-Mok;Choi, Young-Soo;Kim, Kyung-Ho;Im, Geon-Il;Moon, Seung-Hyug;Jeong, Sung-Whan;Kim, Hyeon-Tae;Uh, Soo-Taek;Kim, Yong-Hun;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.3
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    • pp.348-358
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    • 1996
  • Background : Interleukin-5 (IL-5) is responsible for eosinophilia in allergic diseases. In allergic bronchial asthma, there is a correlation between the extent of eosinophil infiltration in bronchial mucosa and IL-5 concentrations. In addition, IL-2 concentration is elevated in the airways and associated with eosinophilia in symptomatic patients with bronchial asthma. In animal studies, IL-2 can induce eosinophilia by increasing the synthesis of IL-5, however, it is still unknown how IL-2 can induce eosinophila in human being. The aim of this study is to evaluation the effect and mechanism of IL-2 on prolongation of eosinophil survival. Methods : After purifiing the eosinophils from the venous blood of allergic patients with eosinophilia, we measured the survival rates of eosinophils using trypan blue dye exclusion test, and the number of eosinophils with Randolp's solution. We compared the survival rates of eosinophils in the presence of IL-2 or IL-5. Neutralizing antibody for IL-5 was added in IL-2 treated eosinophils to reveal whether IL-2 induced prolongation of eosinophil survival was mediated by IL-5. We checked IL-5 m-RNA expression of lymphocytes in the presence of IL-2 by using Reverse transcription-Polymerase chain reaction (RT-PCR) method to revealed the effect of IL-2 on IL-5 m-RNA expression on lymphocyte. $\alpha$ and $\beta$ IL-2 receptors were measured on eosinophils and lymphocytes with flow-cytometer after stimulated with IL-2. Results : 1) Eosinophil survival rates increased dose dependently on IL-5 and IL-2. 2) The eosinophil survival rates increased by IL-2 were not inhibited by the pretreatment with neutralizing antibody for IL-5. 3) IL-5 m-RNA was not expressed on lymphocytes by the treatment with IL-2 up to 96 hours. 4) IL-2 upregulate the expression of IL-$2R{\alpha}$ on eosinophils, instead of no effect on the expression of IL-$2R{\beta}$. Conclusion: Interleukin-2 had the enhancing effect on the survival rates of eosinophils. The mechanism behind IL-2 induced eosinophilia might be the increment of IL-2 receptors on eosinophils rather than IL-5 synthesis by lymphocytes.

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Lung Complications After Allogenic Bone Marrow Transplantaion (동종골수이식 후 폐합병증)

  • JeGal, Yang-Jin;Lee, Je-Hwan;Lee, Kyoo-Hyung;Kim, Woo-Kun;Shim, Tae-Sun;Lim, Chae-Man;Koh, Youn-Suck;Lee, Sang-Do;Kim, Woo-Sung;Kim, Won Dong;Kim, Dong-Soon
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.2
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    • pp.207-216
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    • 2000
  • Background : The occurrence of lung complications after allogenic bone marrow transplantation(BMT) has been reported as 40-60 percent. The risk factors for lung complications are whole body irradiation, high dose chemotherapy, graft versus host disease, old age and CMV infection. The prevalence of graft versus host disease is less in Korea than in Western countries, but frequency of CMV infection is higher. Therefore, the pattern of lung complications may be different in Korea from those in Western countries. Methods : A retrospective cohort study was performed on one hundred consecutive adult patients who underwent allogenic bone marrow transplantation from December, 1993 to May, 1999 at Asan Medical Center. Lung complications were divided into two groups by the time of development, within 30days (pre-engraftment) and beyond 30 days (post-engraftment), and then subdivided into infectious and non-infectious complication. Infectious complications were defined as having the organism in blood, BAL fluid, pleural fluid or sputum, or compatible clinical findings in patients, which improved with antibiotics or an anti-fungal therapy. Result: 1) Eighty three episodes of lung complications had occurred in 54 patients. 2) Within thirty days after BMT, non-infectious complications were more common than infections, but this pattern was reversed after 30 days. After one year post-BMT, there was no infectious complication except in cases of recurrence of underlying disease or development of chronic GVHD. 3) Among the non-infectious complications, pleural effusion (27 episodes) was most common, followed by pulmonary edema (8 episodes), bronchiolitis obliterans(2 episodes), diffuse alveolar hemorrhage (1 episode) and bronchiloitis obliterans with organizing pneumonia (1 episode). 4) The infectious complications were pneumonia (bacterial: 9 episodes, viral: 4 episodes, fungal : 5 episodes, pneumocystis carinii : 1 episode), pulmonary tuberculosis(3 episodes) and tuberculous pleurisy (3 episodes). 5) Lung complications were more frequent in CMV positive patients and in patients with delayed recovery of neutrophil count. 6) The mortality was higher in the patients with lung complications. Conclusion : Lung complications developed in 54% after allogenic BMT and were associated with higher mortality.

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Assessment of the Specificity of A Hybridization of Surfactant Protein A by Addition of Non-specific Rat Spleen RNA (Surfactant Protein A mRNA을 이용한 유전자 재결합 반응에서 비특이성 RNA의 첨가에 의한 특이성 검정)

  • Kim, Byeong Cheol;Kim, Mi Ok;Kim, Tae-Hyung;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.4
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    • pp.393-404
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    • 2004
  • Background : Nucleic acid hybridization has become an essential technique in the development of our understanding of gene structure and function. The quantitative analysis of hybridization has been used in the measurement of genome complexity and gene copy number. The filter hybridization assay is rapid, sensitive and can be used to measure RNAs complementary to any cloned DNA sequence. Methods : The authors assessed the accuracy, linearity, correlation coefficient and specificity of the hybridization depending on the added dose(0, 1, 5, and $10{\mu}g$) of non-specific rat spleen RNA to hybridization of surfactant protein A mRNA. Filter hybridization assays were used to obtain the equation of standard curve and thereby to quantitate the mRNA quantitation. Results : 1. Standard curve equation of filter hybridization assay between counts per minute (X) and spleen RNA input (Y) was Y=0.13X-19.35. Correlation coefficient was 0.98. 2. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) was Y=0.00066X-0.046. Correlation coefficient was 0.99. 3. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $1{\mu}g$ spleen RNA was Y=0.00056X-0.051. Correlation coefficient was 0.99. 4. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $5{\mu}g$ spleen RNA was Y=0.00065X-0.088. Correlation coefficient was 0.99. 5. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $10{\mu}g$ spleen RNA was Y=0.00051X-0.10. Correlation coefficient was 0.99. Conclusions : Comparison of cpm/filter in a linear range allowed accurate and reproducible estimation of surfactant protein A mRNA copy number irrespective of the addition dosage of non-specific rat spleen RNA over the range $0-10{\mu}g$.

Phase II Study of Induction Irinotecan + Cisplatin Chemotherapy Followed by Concurrent Irinotecan + Cisplatin Plus Twice-Daily Thoracic Radiotherapy (유제한성 병기의 소세포 폐암에서 3주 간격으로 시행된 irinotecan과 cisplatin을 이용한 과다분할 방사선 동시 요법)

  • Lee, Jeong Eun;Park, Hee Sun;Jung, Sung Soo;Kim, Ju Ock;Cho, Moon June;Kim, Jin Hwan;Lee, Choong Sik;Kim, Sun Young
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.2
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    • pp.154-164
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    • 2007
  • Background: Irinotecan hydrochloride, a topoisomerase I inhibitor, is effective against small-cell lung cancer. Irinotecan also can act as a potential radiation sensitizer along with cisplatin. To evaluate efficacy and toxicity of irinotecan plus cisplatin (IP) with concurrent thoracic radiotherapy, we conducted a phase II study of IP followed by concurrent IP plus hyperfractionated thoracic radiotherapy in patients with previously untreated limited-stage small-cell lung cancer. Methods: Twenty-four patients with previously untreated small-cell lung cancer were enrolled onto the study since November 2004. Irinotecan $60mg/m^2$ was administered intravenously on days 1 and 8 in combination with cisplatin $60mg/m^2$ on day1 every 21 days. From the first day of third cycle, twice-daily thoracic irradiation (total 45 Gy) was given. Prophylactic cranial irradiation was given to the patients who showed complete remission after concurrent chemoradiotherapy. Restaging was done after second and sixth cycle with chest CT and/or bronchosocpy. Results: Up to November 2004, 19 patients were assessable. The median follow-up time was 12.5 months. A total of 99 cycles (median 5.2 cycles per patient) were administered. The actual dose intensity values were cisplatin $19.6mg/m^2$/week and irinotecan $38.2mg/m^2$/week. Among the 19 patients, the objective response rate was 95% (19 patients), with 9 patients (47%) having a complete response (CR). The major grade 3/4 hematological toxicities were neutropenia (35% of cycles), anemia (7% of cycles), thrombocytopenia (7% of cycles). Febrile neutropenia was 4% of cycles. The predominant grade 3/4 non-hematological toxicities was diarrhea (5% of cycles). Toxicities was not significantly different with concurrent administration of irinotecan and cisplatin with radiotherapy, except grade 3/4 radiation esophagitis (10% of patients). No treatment-related deaths were observed. The 1-year and 2-year survival rate of eligible patients was 89% (16/18) and 47% (9/18), respectively. Conclusion: Three-week schedule of irinotecan plus cisplatin followed by concurrent IP plus hyperfractionated thoracic radiotherapy is an effective treatment for limited disease small-cell lung cancer, with acceptable toxicity.

Effects of Quinone Reductase Induction and Cytotoxicity of the Angelica radix Extracts (당귀 추출성분의 세포독성 효과와 Quinone Reductase 유도활성 효과)

  • 배송자;한은주;노승배
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.1
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    • pp.147-152
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    • 2000
  • Various lines of evidence suggest that dietary components protect the initiation of carcinogenesis. In this study, the ethanol extracts (AGE) and the methanol and hexane partition layers (AGEM, AGEH) of the Angelica radix were screened for their cytotoxic effects using the MTT assay on HepG2, HeLa, MCF7 and SW626 cells and for their ability to induce quinone reductase (QR) in HepG2 cells. AGEM and AGEH of the Angelica radix showed the strongest cytotoxic effects on HepG2 and HeLa cells. Cell growth was inhibited by 99.8% and 99.8% on HepG2 cells and 99.3% and 99.4% on HeLa cells, at dose of $100\;\mu\textrm{g}/ml$ of AGEM and AGEH extracts respectively. AGE and AGEH significantly induced QR activities in the HepG2 cells. The QR activities of HepG2 cells grown in the presence of AGE, AGEH, and AGEM at the concentration of $50\;\mu\textrm{g}/mL$ were 313.5, 273.3 and 133.3 nmol/min/mg protein, respectively. Therefore, based on these studies, Angelica radix may be developed into a potentially useful cancer chemopreventive agent.

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Antioxidant and Anticancer Activities of Water Extracts from Ceramium kondoi (비단풀 물 추출물의 항산화력 및 항암활성)

  • An, Deok-Ho;Cho, Seug-Ja;Jung, Eun-Sil;Lee, Hyun-Jin;Hwang, Ji-Hwan;Park, Eun-Ju;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.10
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    • pp.1304-1308
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    • 2006
  • The present study describes the preliminary evaluation of the antioxidant activity and the cytotoxic effect of Ceramium kondoi. The antioxidant activities and cytotoxic effect of the water extracts were evaluated by total phenolic contents (TPC), DPPH radical scavenging activity (RSA), reducing power (RP), comet assay, and MTT reduction assay. TPC, DPPH RSA, and RP of the extract at the concentration of $1,000{\mu}g/mL$ was $659.2{\mu}M$, 86.0%, and 1.084, respectively, and those were concentration dependent. The $200{\mu}M\;H_2O_2-induced$DNA damage was inhibited by C. kondoi water extract in a dose dependent manner in human leukocytes. The inhibition was by 62.3, 39.8, 24.8% and 16.4% at the concentration of 5, 10, $25{\mu}g/mL$ and $50{\mu}g/mL$, respectively. Cytotoxic activity on HT-29 cells and MCF-7 cells of the C. kondoi water extract at the concentration of $10{\mu}g/mL$ was 49% and 60%, respectively. These results strongly support the possibility of C. kondoi as a source of natural functional materials.