• Title/Summary/Keyword: over-expression

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Identification and Functional Analysis of Escherichia coli RNase E Mutants (Escherichia coli 리보핵산 내부분해효소 RNase E의 돌연변이체 선별 및 특성분석)

  • Shin, Eun-Kyoung;Go, Ha-Young;Kim, Young-Min;Ju, Se-Jin;Lee, Kang-Seok
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.325-330
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    • 2007
  • RNase E is an essential Escherichia coli endoribonuclease that plays a major role in the decay and processing of a large fraction of RNAs in the cell and expression of N-terminal domain consisted of 1-498 amino acids (N-Rne) is sufficient to support normal cellular growth. By utilizing these properties of RNase E, we developed a genetic system to screen for amino acid substitutions in the catalytic domain of the protein (N-Rne) that lead to various phenotypes. Using this system, we identified three kinds of mutants. A mutant N-Rne containing amino acid substitution in the S1 domain (I6T) of the protein was not able to support survival of E. coli cells, and another mutant N-Rne with amino acid substitution at the position 488 (R488C) in the small domain enabled N-Rne to have an elevated ribonucleolytic activity, while amino acid substitution in the DNase I domain (N305D) only enabled N-Rne to support survival of E. roli cells when the mutant N-Rne was over-expressed. Analysis of copy number of ColEl-type plasmid revealed that effects of amino acid substitution on the ability of N-Rne to support cellular growth stemmed from their differential effects on the ribonucleolytic activity of N-Rne in the cell. These results imply that the genetic system developed in this study can be used to isolate mutant RNase E with various phenotypes, which would help to unveil a functional role of each subdomain of the protein in the regulation of RNA stability in E. coli.

THE ANTICANCER EFFECT OF PACLITAXEL($Taxol^{(R)}$) IN ORAL SQUAMOUS CELL CARCINOMA XENOGRAFT (이종 이식된 구강편평세포 암종에서 Paclitaxel ($Taxol^{(R)}$)의 항암 효과)

  • Kim, Ki-Hwan;Kim, Chul-Hwan;Han, Se-Jin;Lee, Jae-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.2
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    • pp.95-110
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    • 2006
  • The treatment for oral and maxillofacial carcinoma with chemotherapeutic agents is evaluated by many effective methods to reduce the tumor mass and cancer cell proliferation. However these chemotherapy have many serious side effects, such as bone marrow suppression, renal toxicity, G-I troubles. Therefore a possible approach to develop a clinically applicable chemotherapeutic agent is to screen anticancer activity of Taxol which is known to have very little side effect and have been used to breast cancer and ovarian carcinoma. Taxol is a new anti-microtubular anti-cancer agent extracted from the bark of the Pacific yew, Taxus brevifolia. Paclitaxel(Taxol) acts by promoting tubulin polymerization and over stabilizing microtubules agianst depolymerization. Despite the constant improvements of methods of the cancer treatment especially chemotherapy, the rate of cancer metastasis and recurrent are not decreased. Thus the investigation of new drug which have very little side effect and a possible clinically application continues to be a high priority. Considering that the Taxol have shown very effective chemotherapeutic agent with relatively low toxicity in many solid tumors, it deserves to evaluate its efficacy in oral squamous cell carcinoma. In this study, to investigate the in-vivo and in-vitro anti-cancer efficacy of Taxol in oral squamous cell carcinoma and lastly, the potency of Paclitaxel in the clinical application for oral cancer was evaluated. In vivo study, after HN22 cell line were xenografted in nude mice, the growth of tumor mass was observed, 3 mg/Kg taxol was injected intraperitoneally into nude mice containing tumor mass. The methods of these study were measurement of total volume of tumor mass, histopathologic study, immunohistochemical study, drug resistance assay, growth curve, MTT assay, flow cytometry, cDNA microarray in vivo and in vitro. The results were obtained as following. 1. The visual inspection of the experimental group showed that the volume of the tumor mass was slightly decreased but no significant difference with control group. 2. Ki-67 index was decreased at weeks 4 in experimental group. 3. Microscopic view of the xenografted tumor mass showed well differentiated squamous cell carcinoma and after Taxol injection, some necrotic tissue was seen weeks 4. 4. The growth curve of the tumor cells were decreased after 1day Taxol treatment. 5. According to the MTT assay, HN22 cell line showed relative drug resistancy above $5\;{\mu}g/ml$ concentrations of Taxol. 6. In drug resistance assay, the decrease of cell counts was seen relatively according to concentration. 7. In Flow cytometry, G2M phase cell arrests were seen in low concentration of the Taxol, while S phase cell arrests were seen in high concentration of the Taxol. 8. Using cDNA microarray technique, variable gene expression of ANGPTL4, TXNRD1, FAS, RRAGA, CTGF, CYCLINEA, P19, DUSP5, CEBPG, BTG1 were detacted in the oral squamous cell carcinoma cell after taxol treatment. In this study paclitaxel is effective against oral squamous cell carcinoma cell lines in vitro, but week effect was observed in vivo. So we need continuous study about anticancer effect of taxol in vivo in oral squamous cell carcinoma.

Aberrant Methylation of p16 Tumor Suppressor Gene and Death-Associated Protein Kinase in Non-Small Cell Lung Carcinoma (비소세포폐암 조직에서 p16 종양억제유전자와 Death-Associated Protein Kinase의 Aberrant Methylation의 양상)

  • Kim, Yun-Seong;Lee, Min-Ki;Jung, Kyung-Sik;Kim, Ki-Uk;Kim, Young-Dae;Lee, Hyung-Ryul;Lee, Chang-Hoon;Seok, Ju-Won;Kim, Yong-Ki;Jun, Eun-Sook;Choi, Young-Min;Rha, Seo-Hee;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.51 no.2
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    • pp.108-121
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    • 2001
  • Background : The $p16^{INK4a}$ (p16) twnor suppressor gene is frequently inactivated in hwnan non-small cell lung cancers (NSCLCs), predominantly through homozygous deletion or in association with aberrant promotor hypermethylation. Death-associated protein kinase (DAPK) gene influences interferon $\gamma$-induced apoptotic cell death and has important role in metastasis of lung cancer in animal model. Hypermethylation of promoter region of DAP kinase gene may suppress the expression of this gene. Methods : This study was performed to investigate the aberrant methylation of p16 or DAP kinase in 35 resected primary NSCLCs by methylation-specific PCR (MSP), and demonstrated frequency, diagnostic value and clinical implication of aberrant methylation of two genes. Results : Thirty-two cases were male patients, and 3 cases were female patients with an average age was 57. $8{\pm}10.5$ years. The histologic types of lung cancer were 22 of squamous cell carcinoma, 12 of adenocarcinoma, 1 of large cell carcinoma. Pathologic stages were 11 cases of stage I (1 IA, 10 IB), 13 cases of stage II (1 IIA, 12 IIB), and 11 cases of stage III (9 IIIA, 2 IIIB). Regarding for the cancer tissue, p16 aberrant methylation was noted in 13 case of 33 cases (39.4%), DAP kinase in 21 cases of 35 cases (60%). Age over 55 year was associated with p16 aberrant methylation significantly (p<0.05). Methylation status of two genes was not different by smoking history, histologic type, size of tumor, lymph node metastasis and disease progression of lung cancer. There was no correlation between p16 and DAP kinase hypermethylation. Conclusion: This investigation demonstrates that aberrant methylation of p16 tumor suppressor gene or DAP kinase showed relatively high frequency (74.3%) in NSCLCs, and that these genes could be a biologic marker for early detection of lung cancer.

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A Study on Soviet Constructive Fashion in 1920s (1920년대 소비에트 구성주의 패션에 관한 연구)

  • 조윤경;금기숙
    • Journal of the Korean Society of Costume
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    • v.36
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    • pp.183-203
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    • 1998
  • The wave of Avant-garde swept away all in the unique social background so called 'October Revolution' and the early 1900 Russian society which was able to absorb and accept anything. The Russian avant-garde has been affected by the Cubism and the Futurism those had peculiarly appeared in the early twentieth century, spreaded out to three spheres: the Suprematism, the Rayonism and the Constructivism. The Russian Constructivism has appeared in this background, concretely and ideally ex-pressed the ideology of the revolution into the artistic form and made an huge influence to the whole Russian society. The Constructivist like Tatlin, naum Gabo, Pevaner, Rodchenko, Stepanova, Popova and Exter gave great effect on the Soviet Constructive fashion design in 1920's after the Revolution. The Soviet costume in 1920s hold in common the characteristicss of the Constructive graphic as it is, geometrical and abstractive form, energetic and motility. In fashion design, these graphic qualities have been showed as the application of geometrical form and architectural image, physical distortion and transformation. And in textile design, the simple, dynamical presentation has been appeared. We can classify the Soviet costume at this time into three occasions. The first term is from late 1910 th mid 1920, and it is altered from folk costume design to modern one. With Lamanova as the first on the list, using the folk mitif, the Constructive expression of simple form has been gradually revealed in design. Designers like Makarova, Pribylskaia and Mukhina produced the plane, simple chemise style with the decoration of the Russian traditional motif. From early to late 1920 is the second term, and it is at the pick of the most active processing of the Constructive design. Not only at the costume in daily life but also at the theatrical costume and textile, the con-structive design has been represented all avail-able fields. Many Constructivists including Stepanova, Popova, Exter and Rodchenko took part in the textile design and costume design so as to evlvo their aesthetic concept. The third term is from late 1920 to early 1930. The socialistic realism has dominated over the whole culture and art, the revolutionary dynamic motif has been presented also in textile design. The formative features of Soviet Constructive fashion design are; silhouette, from, motif, color and fabric. The first, the silhouette : a straight rectangular silhouetted has been expressed through the whole period and a volumed one with distorted human body shape has introduced in the theatrical costume design. The second, the form: many lengthened rectangular forms have been made at beginnings, but to the middle period, geometrical, architectural forms have been more showed and there are energy and movement in design. At the last period, only a partial feature-division has been seen. The third, the motif; no pattern or ethnic motif has been partly used at beginnings, a figure like circle, tri-angle has gradually appeared in textile design. At latter period, a real-existent motif like an airplane has been represented with graphing and simplicity. The fourth, the color ; because of insufficient dyeing, neutral color like black or grey color has been mainly covered, but after middle term, a primary color or pastel tone has been seen, contrast of the fabric; without much development of textile industry after the Revolution, thick and durable fabrics have been the main stream, but as time had going to the last period, fabrics such as linen, cotton, velvet and silk have been varously choesn. At the theatrical costume, new materials like plastics and metals that were able to accentuate the form. The pursuit of popularity, simplicity and functionalism that the basic concept of Constructive fashion is one of the "beauty" which has been searching in modern fashion. And now we can appreciate how innovative and epochal this Soviet Constructive fashion movement was.ement was.

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Prediction of Target Motion Using Neural Network for 4-dimensional Radiation Therapy (신경회로망을 이용한 4차원 방사선치료에서의 조사 표적 움직임 예측)

  • Lee, Sang-Kyung;Kim, Yong-Nam;Park, Kyung-Ran;Jeong, Kyeong-Keun;Lee, Chang-Geol;Lee, Ik-Jae;Seong, Jin-Sil;Choi, Won-Hoon;Chung, Yoon-Sun;Park, Sung-Ho
    • Progress in Medical Physics
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    • v.20 no.3
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    • pp.132-138
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    • 2009
  • Studies on target motion in 4-dimensional radiotherapy are being world-widely conducted to enhance treatment record and protection of normal organs. Prediction of tumor motion might be very useful and/or essential for especially free-breathing system during radiation delivery such as respiratory gating system and tumor tracking system. Neural network is powerful to express a time series with nonlinearity because its prediction algorithm is not governed by statistic formula but finds a rule of data expression. This study intended to assess applicability of neural network method to predict tumor motion in 4-dimensional radiotherapy. Scaled Conjugate Gradient algorithm was employed as a learning algorithm. Considering reparation data for 10 patients, prediction by the neural network algorithms was compared with the measurement by the real-time position management (RPM) system. The results showed that the neural network algorithm has the excellent accuracy of maximum absolute error smaller than 3 mm, except for the cases in which the maximum amplitude of respiration is over the range of respiration used in the learning process of neural network. It indicates the insufficient learning of the neural network for extrapolation. The problem could be solved by acquiring a full range of respiration before learning procedure. Further works are programmed to verify a feasibility of practical application for 4-dimensional treatment system, including prediction performance according to various system latency and irregular patterns of respiration.

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Clinical Characteristics and Genetic Analysis of Prader-Willi Syndrome (Prader-Willi 증후군의 임상 양상 및 유전학적 진단에 관한 고찰)

  • Lee, Ji Eun;Moon, Kwang Bin;Hwang, Jong Hee;Kwon, Eun Kyung;Kim, Sun Hee;Kim, Jong Won;Jin, Dong Kyu
    • Clinical and Experimental Pediatrics
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    • v.45 no.9
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    • pp.1126-1133
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    • 2002
  • Purpose : Prader-Willi syndrome(PWS) is a complex disorder affecting multisystems with characteristic clinical features. Its genetic basis is an expression defect in the paternally derived chromosome 15q11-q13. We analyzed the clinical features and genetic basis of PWS patients for early detection and treatment. Methods : We retrospectively studied 24 patients with PWS in Department of Pediatrics, Samsung Medical Center, from September 1997 to September 2001. We performed cytogenetic and molecular genetic techniques using high resolution GTG banding techniques, fluorescent in situ hybridization and methylation-specific PCR for CpG island of SNRPN gene region. Results : The average birth weight of PWS patients was $2.67{\pm}0.47kg$ and median age at diagnosis was 1.3 years. The average height and weight of PWS patients under one year at diagnostic time were located in a 3-10 percentile relatively, and a rapid weight gain was seen between two and six years. Feeding problems in infancy and neonatal hypotonia were the two most consistently positive major criteria in over 95% of the patients. In 18 of the 24 cases(75%), deletion of chromosome 15q11-q13 was demonstrated and one case among 18 had an unbalanced 14;15 translocation. In four cases without any cytogenetic abnormality, it may be considered as maternal uniparental disomy and the rest showed another findings. Conclusion : We suggest diagnostic testing for PWS in all infants/neonates with unexplained feeding problems and hypotonia. It is necessary for clinically suspicious patients to undergo an early genetic test. As the genetic basis of PWS was heterogenous and complex, further study is required.

Anti-inflammatory Activity of Crinum asiaticum Linne var. Japonicum Extract and its Application as a Cosmeceutical Ingredient (문주란의 항염효과와 화장료적 특성)

  • Kim, Ki-Ho;Kim, Young-Heui;Kim, Ki-Soo;Park, Sun-Hee;Lee, Soo-Hee;Kim, Young-Jin;Kim, Young-Sil;Kim, Jong-Heon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.1 s.55
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    • pp.59-64
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    • 2006
  • Crinum asiaticum Linne var. japonicum has long been used as a rheumatic remedy, an anti-pyretic, an anti-ulcer treatment, and for the alleviation of local pain and fever in Korea and Malaysia. In order to investigate the possibility of Crinum asiaticum Linne var. japonicum extract as a cosmetic ingredient, we measured its anti-inflammatory effect by inhibition of iNOS (inducible nitric oxide synthase), and the release of PGE2, IL-6, and IL-8. HPLC experiment after extraction with 95% ethanol at pH 3.5 showed that Crinum asiaticum Linne var. japonicum was mainly composed of lycorine (up to 1%), a well-known immunosuppressant. The content of lycorine varied depending on the type of tissue analyzed and the extraction method. In anti-inflammatory assay for inhibition of nitric oxide formation on lipopolysaccharide (LPS)- activated mouse macrophage RAW 264.7 cells, the ethanolic extract of Crinum asiaticum showed inhibitory activity of NO production in dose-dependent manner ($IC_{50} = 83.5 {\mu}g/mL$). Additional study by RT-PCR demonstrated that the extract of Crinum asiaticum significantly suppressed the expression of the iNOS gene. Moreover, the extract of Crinum asiaticum did not show my cytotoxicity, but did show cell proliferation effect against LPS ($10{\sim}60%$ increase of tell viability). In an assay to determine inhibition of the $H_2O_2$-activated release of PGE2, IL-6, and IL-8 in human normal fibroblast cell lines, the release of PGE2 and IL-6 was almost completely inhibited above concentrations of 0.05% and 1%, respectively. Moreover, the release of IL-8 was completely inhibited over the entire range of concentrations (> 0.0025%). The result showed that the extract of Crinum asiaticum Linne var. japonicum has sufficient anti-inflammatory effect. There-fore, Crinum asiaticum Linne var. japonicum extract may be useful as an ingredient of cosmetic products.

The Neuroprotective Effects of 6-cyano-7-nitroquinoxalin-2,3-dione (CNQX) Via Mediation of Nitric Oxide Synthase on Hypoxic-ischemic Brain Injury in Neonatal Rats

  • Jung, Ji-Eun;Keum, Kyung-Hae;Choi, Eun-Jin;Kim, Jin-Kyung;Chung, Hai-Lee;Kim, Woo-Taek
    • Neonatal Medicine
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    • v.18 no.1
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    • pp.59-69
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    • 2011
  • Purpose: Current studies have demonstrated the neuroprotective effects of 6-cyano-7-nitroquinoxalin-2,3-dione (CNQX) in many animal models of brain injury, including hypoxic-ischemic (HI) encephlopathy, trauma and excitotoxicity, but limited data are available for those during the neonatal periods. Here we investigated whether CNQX can protect the developing rat brain from HI injury via mediation of nitric oxide synthase. Methods: In an in vivo model, left carotid artery ligation was done in 7-day-old Sprague-Dawley (SD) rat pups. The animals were divided into six groups; normoxia (N), hypoxia (H), hypoxia with sham-operation (HS), hypoxia with operation (HO), HO treated with vehicle (HV), and HO treated with CNQX at a dose of 10 mg/kg (HC). Hypoxia was made by exposure to a 2 hr period in the hypoxic chamber (92% $N_2$, 8% $O_2$). In an in vitro model, embryonic cortical neuronal cell culture of SD rats at 18-day gestation was done. The cultured cells were divided into three groups: normoxia (N), hypoxia (H), and hypoxia treated with CNQX (HC). The N group was prepared in 5% $CO_2$ incubators and the other groups were placed in 1% $O_2$) incubators (94% $N_2$, 5% $CO_2$) for 16 hr. Results: In the in vitvo and in vivo models, the expressions of iNOS and eNOS were reduced in the hypoxia group when compared to the normoxia group, whereas they were increased in the CNQX-treated group compared to the hypoxia group. In contrast, the expression of nNOS was showed reversely. Conclusion: CNQX has neuroprotective property over perinatal HI brain injury via mediation of nitric oxide synthase.

Therapeutic Use of Music for Stuttering Children (말더듬 아동을 위한 음악치료적 접근)

  • Cho, Jung Min
    • Journal of Music and Human Behavior
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    • v.4 no.1
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    • pp.21-30
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    • 2007
  • Unlike other common forms of speech disorder, such as phonological disorder or dysphonia, stuttering has not been studied within the context of music therapy. Most cases of stuttering display no difficulty in singing, and fluency within the musical structure does not translate to fluency in speech. Hence, musical approach has been generally considered to be ineffective to the treatment of stuttering. However, the fundamentals of music therapy assume its extensive application in treating variety of speech disorders, including the case of stuttering. Presented in this paper are the case studies designed to validate the efficacy of music therapy as a remedy for stuttering. This study enrolled 6 children with stuttering and conducted 20 individual sessions over a period of 10 weeks. The sessions focused on the Melodic Intonation Therapy, Reinforcement of speech rhythm, song writing and singing. Musical elements were structured to enhance the verbal expression and rhythmic senses, as well as to facilitate the initiation of verbal communication. The result is as follows. First, it was noticed that the disfluency had been decreased in before and after of the music therapy in every child although the result was somewhat different depending the child. The overall result of the investigation shows the significant difference statistically. And categorically speaking, the significant difference was checked in the frequency of the stuttering. In the steps of the session, the increase and decrease was happened repeatedly, and then after it was decreased little by little. Secondly, the Communication Attitude was decreased in before and after of the music therapy, and also there was significant difference statistically. although the avoidance behavior was decreased in before and after of the music therapy, the increase and the decrease was repeated irregularly in the steps of session. All the results described above shows that music therapy gives positive effect to decrease in disfluency of stuttering child and also to develop the Communication Attitude. And new possibility and effectiveness can be proposed in the musical approach to the stuttering.

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The Anti-inflammatory Effect of Skipjack Tuna (Katsuwonus pelamis) Oil in LPS-induced RAW 264.7 Cells and Mouse Models (LPS 유도 RAW 264.7 세포와 마우스 모델에서 참치(Katsuwonus pelamis) 유의 항염증 효과)

  • Kang, Bo-Kyeong;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Ahn, Na-Kyung;Choi, Yeon-Uk;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Park, Ji-Hye;Bae, Nan-Young;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.45-55
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    • 2015
  • This study was carried out to demonstrate the anti-inflammatory effect of tuna oil (TO) using LPS-induced inflammation responses and mouse models. First, nitric oxide (NO) and pro-inflammatory cytokines levels were suppressed up to 50% with increasing concentrations of TO without causing any cytotoxicity. Also, the expression of a variety of proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and nuclear factor kappa B (NF-κB), was suppressed in a dosedependent manner by treatment with TO. Furthermore, TO also inhibited the phosphorylation of mitogen-activated protein kinases (MAPKs), including c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 protein kinase (p38). Moreover, in in vivo testing the formation of ear edema was reduced at the highest dose tested compared to that in the control, and a reduction of ear thickness and the number of mast cells was observed in histological analysis. In acute toxicity test, no mortalities occurred in mice administrated 5,000 mg/kg body weight of TO over a two-week observation period. Our results suggest that TO has a considerable anti-inflammatory property through the suppression of inflammatory mediator productions and that it could prove to be useful as a potential anti-inflammatory therapeutic material.