• Title/Summary/Keyword: enzyme assay

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Ehrlichia and Borrelia spp. Infection in German Shepherd Dogs in Korea (독일 셰퍼드 개에서 Ehrlichia와 Borrelia spp.의 감염)

  • Hwang, Cheol-Yong;Seo, Sang-Ho;Kang, Jun-Gu;Youn, Hwa-Young;Chae, Joon-Seok
    • Journal of Veterinary Clinics
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    • v.28 no.2
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    • pp.204-210
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    • 2011
  • The presence of the tick-borne pathogens Ehrlichia and Borrelia in German Shepherd dogs in Korea was determined by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). A total of 291 dogs were randomly selected from five Korean provinces from October 2005 through September 2006. The seroprevalence of antibodies to canine Ehrlichia and Borrelia agents detected by ELISA (Snap$^{(R)}$ 3Dx$^{(R)}$ Test, IDEXX Laboratories) was 7.56% (22 dogs) and 1.72% (5 dogs) respectively, throughout the country. Positive antibodies against both pathogens were detected in two dogs (0.69%). The provincial distribution of seroprevalence against Ehrlichia was 1.28% (1 of 78) in Gyeonggi-do, 12.64% (11 of 87) in Gangwon-do, 9.76% (4 of 41) in Chungcheong-do, 8.93% (5 of 56) in Gyeongsang-do, and 3.45% (1 of 29) in Jeolla-do. According to PCR analysis, Ehrlichia chaffeensis target DNA was amplified in 3.09% (9 of 291 dogs) of blood samples, 2.41% (7 of 291) from Gangwon-do and 0.69% (2 of 291) from Chungcheong-do. The oligonucleotide sequences (SNU-EC3 and SNU-EC5) from the PCR fragment examined in Korea were closely related to E. chaffeensis isolated from the tick Haemaphysalis longicornis, in China and the state of Arkansas in the US. Based on these results, the presence of E. chaffeensis infection was identified in German Shepherds being bred in Korea. These results bring to light the importance of paying close attention to tick-borne infections such as Lyme disease during clinical diagnosis. This infectious disease should be included as a differential diagnosis for patients who participate in outdoor activity from spring to fall or who have thrombocytopenia or leucopenia.

Distinct Spatio-temporal Expression Patterns of Patatin Promoter-GUS Gene Fusion in Transgenic Potato Microtubers (형질전환 감자 소괴경의 발달단계에 따른 Patatin Promoter-GUS 유전자의 발현 분석)

  • Youm, Jung-Won;Kim, Mi-Sun;Lee, Byoung-Chan;Kang, Won-Jin;Jeon, Jae-Heung;Joung, Hyouk;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.13-18
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    • 2003
  • This study was carried out to investigate the expression patterns of foreign gene that controlled by tuber-specific patatin promoter in transgenic potatoes. Potato leaf disc cultured in vitro were transformed by the Agrobacterium strain LBA4404 containing pBl121 or pATGUS from potato cv. Desiree. In order to select the transgenic lines, gene-specific primers deduced from the NPTII were synthesized and used for polymerase chain reaction. The down part of the putative transgenic potatoes was transplanted weekly onto sucrose-enriched medium to accelerate the microtuber formation. RNA gel blot analysis was performed on the total RNAs obtained from tuber that had been harvested at a week interval. Also, histochemical assay was observed in the explants transformed with either pBI121 or pATGUS. Results showed that the transgenic plant containing pATGUS expressed GUS transcripts mainly at the tuber, not in stem, with the highest expression level in 5 weeks-grown microtubers. In contrast to pATGUS plants, the transformed plants with pBI121 showed an equal expression pattern throughout the whole developing stages. Consistent with RNA gel blot analysis, histochemical GUS staining and enzyme activity exhibited pATGUS transcripts were at the highest level in 5 weeks cultures. From these results, we suggest that the best stage to analyze the foreign gene introduced by patatin promoter into potato plants is at 5 weeks cultures after tuber formation.

Epstein-Barr Virus Antibodies in Korean Mothers and Their Neonates (산모와 신생아의 Epstein Barr Virus 항체가에 관한 연구)

  • Shin, Young Kyoo;Eun, Baik Lin;Park, Sang Hee;Lim, Chae Seung;Kim, Young Sik
    • Pediatric Infection and Vaccine
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    • v.5 no.1
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    • pp.121-127
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    • 1998
  • Purposes : This study was performed to evaluate the seropositivities and levels of term pregnant women and their neonates, and the transplacental transfer rate of maternal Epstein-Barr Virus-specific IgG(VCA IgG and EBNA IgG) from term pregnant women to their neonates. Subjects and Methods : During Jan. 1, 1997 to Mar. 31. 1997, we collected the 42 pairs of sera from pregnant women and umbilical cord of their neonates in Korea University Ansan Hospital. The serum levels of VCA IgG and EBNA IgG were measured by the ELISA method. Results : 1) The seropositivities of VCA IgG were 100% in mothers and neonates. There was no statistical difference of mean VCA IgG levels between mothers and neonates. There was significant correlation of VCA IgG levels between maternal sera and neonatal umbilical cord sera(correlation coefficient r=0.5214, P<0.001). 2) The seropositivities of EBNA IgG were 100% in mothers and neonates. There was no significant difference of the mean EBNA IgG levels between mothers and neonates. There was significant correlation of EBNA IgG levels between maternal sera and neonatal umbilical cord sera (correlation coefficient r=0.7244, P<0.001). 3) There was no correlation between VCA IgG and EBNA IgG levels of maternal sera. Conclusion : Seropositivities of EBV CA IgG and EBNA IgG of term-pregnant women and their neonates were 100% and no significant differences of antibody levels were found in two groups. It seems that EBV Antibody levels in Korean mothers and neonates were high enough to protect primary EBV infection during early infancy.

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The Clinical Study of the Lower Respiratory Tract Infection by Respiratory Syncytial Virus on Children under 2 Year of Age (2세 이하 어린이에서 Respiratory Syncytial Virus에 의한 하기도 감염에 관한 연구)

  • Cho, Jung Ik;Choi, Hyung Chul;Kim, Jong Duck;Cho, Ji Hyun
    • Pediatric Infection and Vaccine
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    • v.7 no.2
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    • pp.193-200
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    • 2000
  • Purpose : This study was designed and performed for evaluations of clinical manifestation and course of the children under 2 year of age with respiratory tract infection and positive respiratory syncytial virus(RSV) antigen. Methods : The selection criteria of the patients were children under 24 month-of-age, Clinical manifestation of respiratory tract infection, and positive RSV antigen that was detected by Vitek ImmunoDiagnostic Assay System(VIDAS) from nasal cavity. The additional laboratory and simple chest X-ray findings were reviewed from the chart of children who were admitted Wonkwang university hospital from October 1999 to March 2000. Results : Total number of patients enrolled on this study was 102. The 48(47%) children were RSV antigen positive by VIDAS method. Abnormal chest X-ray findings were noticed in 38 cases. The male to female sex ratio of 48 RSV antigen positive cases was 1.2 : 1. The mean and range of age was $10.2{\pm}5.9$ and 1.0~24 months. The peak outbreak of cases was noticed on November, 1999. All of the cases shows coughing but rale was audible in 30 cases(60%). Dyspnea, wheezing, and intercostal retraction were noticed 11(23%), 15(31%), and 10(21%) cases respectively. The most common chest X-ray finding was scattered patch infiltration that was noticed in 30 cases(63%). The mean total white blood cell counts in peripheral blood was $12,608{\pm}4,686/mm^3$. The mean blood level of IgA and IgE were $50.8{\pm}20.9$ and $72.1{\pm}98.3mg/dL$ respectively. The C-reactive protein was $16.0{\pm}18.5mg/L$. Total 5 cases need a mechanical respiraton. The duration of admission was under 7 days in 36 cases(75%). Conclusion : The RSV antigen was detected commonly in late fall and winter season. The severity of children under 2 years old with RSV respiratory tract infection take in some degree a gave courses.

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Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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Diagnostic Value of Serum Procalcitonin in Febrile Infants Under 6 Months of Age for the Detection of Bacterial Infections (발열이 있는 6개월 미만의 영아에서 세균성 감염에 대한 procalcitonin의 진단적 가치)

  • Kim, Nam Hyo;Kim, Ji Hee;Lee, Taek Jin
    • Pediatric Infection and Vaccine
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    • v.16 no.2
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    • pp.142-149
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    • 2009
  • Purpose : The aim of this study was to determine the diagnostic value of serum procalcitonin (PCT) compared with that of C-reactive protein (CRP) and the total white blood cell count (WBC) in predicting bacterial infections in febrile infants<6 months of age. Methods : A prospective study was performed with infants <6 months of age who were admitted to the Department of Pediatrics with a fever of uncertain source between July and September 2008. Spinal taps were performed according to clinical symptoms and physical examination. Serum PCT levels were measured using an enzyme-linked fluorescent assay. Results : Seventy-one infants (mean age, 2.62 months) were studied. Twenty-six infants (36.6%) had urinary tract infections (UTIs), and 22 infants (31.0%) had viral meningitis. The remaining infants had acute pharyngitis (n=1), herpangina (n=1), upper respiratory tract infections (n=7), acute bronchiolitis (n=8), acute gastroenteritis (n=4), and bacteremia (n=2). The median WBC and CRP levels were significantly higher in infants with UTIs than in infants with viral meningitis. However, there were no differences in the median PCT levels between the groups (0.14 ng/mL vs. 0.11 ng/mL, P=0.419). The area under the receiver operating characteristic curve was 0.792 (95% CI, 0.65-0.896) for WBC, 0.77 (95% CI, 0.626-0.879) for CRP, and 0.568 (95% CI, 0.417-0.710) for PCT. An elevated WBC count (>11,920/${\mu}L$) and an increased CRP level (>1.06mg/dL) were significant predictors of UTIs based on multiple logistic regression analysis. Conclusion : Serum PCT concentrations should be interpreted with caution in infants <6 months of age with a fever of uncertain source.

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Influences of Saliva Substitutes on Salivary Enzymatic Activity (타액대체제가 타액 효소 활성에 미치는 영향)

  • Kho, Hong-Seop;Lee, Sung-Woo
    • Journal of Oral Medicine and Pain
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    • v.34 no.3
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    • pp.227-235
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    • 2009
  • Many of the protective functions of saliva can be attributed to the biological, physical, structural, and rheological characteristics of salivary glycoproteins. Therefore, the development of ideal saliva substitutes requires understanding of the rheological as well as biological properties of human saliva. In the present study, we investigated the changes of salivary enzymatic activities by saliva substitutes and compared viscosity of saliva substitutes with human saliva. Five kinds of saliva substitutes such as Moi-Stir, Stoppers4, MouthKote, Saliva Orthana, and SNU were used. Lysozyme activity was determined by the turbidimetric method. Peroxidase activity was determined with an NbsSCN assay. $\alpha$-Amylase activity was determined using a chromogenic substrate, 2-chloro-p-nitrophenol linked with maltotriose. The pH values of saliva substitutes were measured and their viscosity values were measured with a cone-and-plate digital viscometer at six different shear rates. Various types of saliva substitutes affected the activities of salivary enzymes in different ways. Stoppers4 enhanced the enzymatic activities of hen egg-white lysozyme, bovine lactoperoxidase (bLP), and $\alpha$-amylase. Saliva Orthana and SNU inhibited bLP activity and enhanced $\alpha$-amylase activity. MouthKote inhibited $\alpha$-amylase activity. Moi-Stir inhibited the enzymatic activities of bLP and $\alpha$-amylase. The pH values were very different according to the types of saliva substitutes. Stoppers4, MouthKote, and Saliva Orthana showed lower values of viscosity at low shear rates and higher values of viscosity at high shear rates compared with unstimulated and stimulated whole saliva. Moi-Stir and SNU displayed much higher values of viscosity than those of natural whole saliva. Collectively, our results indicate that each saliva substitute has its own biological and rheological characteristics. Each saliva substitute affects the enzymatic activity of salivary enzyme and finally oral health in different ways.

Diagnostic value of C-reactive Protein and Vascular Endothelial Growth Factor in Differentiation of Pleural Effusions (흉막액 감별에 있어서 C-반응성단백과 혈관내피성장인자의 유용성)

  • Kim, Sang Ha;Lee, Won Yeon;Park, Joo Young;Park, Hyun Sook;Han, Hye-Kyoung;Ju, Hun Su;Hong, Tae Won;Lee, Nak Won;Shin, Kye Chul;Yong, Suk Joong
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.5
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    • pp.467-477
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    • 2003
  • Background : Pleural effusions are generally divided into transudates and exudates. If it is exudative, more diagnostic tests are required in order to determine the cause of the local disease. A malignancy is a common and important cause of exudative pleural effusions. Because the pleural fluid cytology and pleural biopsy specimens do not provide a diagnosis in a high percentage of malignant effusions, several tumor markers have been examined. In order to overcome this limitation, this study hypothesized that C-reactive protein(CRP) and vascular endothelial growth factor(VEGF) measurements would be useful for differentiating trasudates from exudates and determining the differences between a benign and malignant effusion. Methods : Eighty consecutive patients with a pleural effusion (tuberculous 20, parapneumonic 20, malignant 20, transudative 20) were examined prospectively: 60 of them were classified according to Light's criteria as having an exudative fluid and 20 had a transudative fluid. The standard parameters of a pleural effusion were examined and the serum and pleural effusion VEGF levels were measured using enzyme linked immunosorbent assay(ELISA). CRP in the serum and pleural fluid was determined by a turbidimetric immunoassay. Results : The pleural CRP levels in the exudates were significantly higher than those in the transudates, $4.19{\pm}4.22mg/d{\ell}$ and $1.29{\pm}1.45mg/d{\ell}$, respectively. The VEGF levels in the pleural effusions were significantly elevated in the exudates compared to the transudate, $1,011{\pm}1,055pg/m{\ell}$ and $389{\pm}325pg/m{\ell}$, respectively. The VEGF ratio in the exudative effusion is significantly higher than a transudative effusions, $3.9{\pm}4.7$ and $1.6{\pm}0.9$, respectively. The pleural CRP levels in the patients with a benign effusion($4.15{\pm}4.20mg/d{\ell}$) were significantly higher than those in the malignant effusion($1.43{\pm}1.91mg/d{\ell}$). The VEGF ratio is significantly higher in malignant effusions($4.9{\pm}5.5$) than in benign effusions($2.8{\pm}3.6$). Conclusion : In conclusion, the CRP and VEGF levels in the serum and pleural effusion can distinguish between transudates and exudates. Moreover it can differentiate between benign and malignant pleural effusions.

Prognostic Utility of the Soluble Triggering Receptor Expressed on Myeloid Cells-1 in Patients with Acute Respiratory Distress Syndrome (급성호흡곤란증후군 환자에서 Soluble Triggering Receptor Expressed on Myeloid Cells-1의 예후인자로서의 유용성)

  • Huh, Jin Won;Jung, Hoon;Lim, Chae-Man;Koh, Younsuck;Hong, Sang-Bum
    • Tuberculosis and Respiratory Diseases
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    • v.65 no.4
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    • pp.301-307
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    • 2008
  • Background: The triggering receptor expressed on myeloid cells-1 (TREM-1) is an activating receptor that is expressed on the surface of neutrophils and mature monocytes when stimulated with several microbial components, which can amplify the inflammatory response. This study analyzed the prognostic value of the sTREM-1 levels in patients with acute respiratory distress syndrome (ARDS). Methods: The bronchoalveolar lavage (BAL) fluid and blood was collected prospectively from 32 patients with ARDS, 15 survivors and 17 nonsurvivors. An enzyme-linked immunosorbent assay was performed to measure the sTREM-1. The following data was obtained: APACHE II score, Clinical Pulmonary Infection Score (CPIS), BAL fluid analysis, C-reative protein. Mortality in the ICU was defined as the end point. Results: The serum sTREM-1 level was significantly higher in the nonsurvivors than survivors ($54.3{\pm}10.3pg/ml$ vs. $22.7{\pm}2.3pg/ml$, p<0.05). The sTREM-1 level in the serum, but not in the BAL fluid, was an independent predictor of the ICU mortality (OR: 22.051, 95% CI: 1.780~273.148, p<0.016), and a cut-off value of ${\geq}33pg/ml$ yielded a diagnostic sensitivity of 71% and specificity of 93%. Conclusion: The serum sTREM-1 level may be a useful predictor of the outcome of ARDS patients.

Antioxidant capacity and Raw 264.7 macrophage anti-inflammatory effect of the Tenebrio Molitor (갈색거저리(Tenebrio Molitor)의 항산화능과 Raw 264.7 대식세포의 항염증 효과)

  • Yu, Jae-Myo;Jang, Jae-Yoon;Kim, Hyeon-Jeong;Cho, Yong-Hun;Kim, Dong-in;Kwon, O-jun;Cho, Yeong-Je;An, Bong-Jeun
    • Food Science and Preservation
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    • v.23 no.6
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    • pp.890-898
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    • 2016
  • The purpose of this paper is to investigate potential anti-inflammatory and anti-oxidant effects of Tenebrio molitor. Macrophage cell response by outside stimulation leads expression of pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin-6 (IL-6), $interleukin-1{\beta}$ ($IL-1{\beta}$), and trigger expression of genes which are affected by inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), resulting in formation of inflammatory factors like nitric oxide (NO) and Prostaglandin $E_2$ (PGE2). Cell viability was determined by MTT assay. In order to investigate anti-inflammatory agents, the inhibitory effects on the production of lipopolysaccharide (LPS)-induced NO in RAW 264.7 cells were examined. T. Molitor significantly decreased the production of NO in a dose-dependent manner, and also reduced the expression of iNOS, a COX-2 protein. As a result, the levels of protein such as $PGE_2$, iNOS, COX-2 and MARKs were significantly reduced compared to non-treated group in T. Molitor water extract (TDW) treated group. Also, antioxidant effect of T. Molitor were investigated using DPPH, ABTS+ and superoxide anion radical scavenging activity tests in cell-free system. Antioxidant activity of T. molitor was found low in the DPPH radical scavenging test while high in the ABTS+ and superoxide anion radical scavenging activity tests. These results show that TDW could be an effective anti-pro-inflammatory and anti-oxidant agent.