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SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT (인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Hwang, Soon-Jeong;Choung, Pill-Hoon;Kim, Myung-Jin;Kim, Nam-Yeol;Yoo, Sang-Bae;Jahng, Jeong-Won;Kim, Hyun-Man;Kim, Joong-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.3
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

NEW ANTIDEPRESSANTS IN CHILD AND ADOLESCENT PSYCHIATRY (소아청소년정신과영역의 새로운 항우울제)

  • Lee, Soo-Jung
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.14 no.1
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    • pp.12-25
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    • 2003
  • Objectives:As increasing number of new antidepressants have been being introduced in clinical practice, pharmacological understanding has been broadened. These changes mandate new information and theories to be incorporated into the treatment process of children with depressive disorders. In light of newly coming knowledge, this review intended to recapitulate the characteristics of new antidepressants and to consider the pivotal issues to develope guidelines for the treatment of depression in childhood and adolescence. Methods:Searching the Pub-Med online database for the articles with the key words of 'new', 'antidepressants' and 'children' ninety-seven headings of review articles were obtained. The author selected the articles of pertinent subjects in terms of either treatment guideline or psychopharmacology of new antidepressants. When required, articles about the clinical effectiveness of individual antidepressants were separatedly searched. In addition, the safety information of new antidepressants was acquired by browsing the official sites of the United States Food and Drugs Administration and Department of Health and Human Services. Results:1) For the clinical course, treatment phase, and treatment outcome, the reviews or treatment guidelines adopted the information from adult treatment guidelines. 2) Systematic and critical reviews unambiguously concluded that selective serotonin reuptake inhibitors(SSRIs) excelled tricyclic antidepressants( TCAs) for both efficacy and side effect profiles, and were recommend for the first-line choice for the treatment of children with depressive disorders. 3) New antidepressants generally lacked treatment experiences and randomized controlled clinical trials. 4) SSRIs and other new antidepressants, when used together, might result in pharmacokinetic and/or pharmacodynamic drug-to-drug interaction. 5) The difference of the clinical effectiveness of antidepressants between children and adults should be addressed from developmental aspects, which required further evidence. Conclusion:Treatment guidelines for the pharmacological treatment of childhood and adolescence depression could be constructed on the basis of clinical trial findings and practical experiences. Treatment guidelines are to best serve as the frame of reference for a clinician to make reasonable decisions for a particular therapeutic situation. In order to fulfill this role, guidelines should be updated as soon as new research data become available.

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Studios on the Metagonimus fluke in the Daecheong Reservoir and the upper stream of Geum River, Borea (대청호 및 그 상류의 Metagonimus 흡충에 관한 연구)

  • 김종환;김남만
    • Parasites, Hosts and Diseases
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    • v.25 no.1
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    • pp.69-82
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    • 1987
  • The prevalences of the cuke belonging to genus Metagonimus hove been reported along the upper stream of inhabitants by several workers since 1980, however the taxonomical problems of the fluke was not yet settled. The larval flukes; cercaria and metacercaria as well as their intermediate hosts, and adult were studied in order to identify the Mepagonimus in the areas. The results obtained are summarized as follows: 1. The snails, Semisulcospira globus were collected (rom the three different localities along the upper stream of the River. The cercariae were found from 125(7.2%) out of 1,730 snails by natural emerging method, and were identified into 5 species including Metagenimus sp. (3.7%), Pseudexorchis major(1.4%), Cercaria nipponensis (0.9%), Cercaria incerpa(0.6%), and Cercaria yoshidae(0.6%). Cercariae of Metagonimus species had four to dye oral spines on its anterior of the first line. 2. The cercariae of Metagonimus were experimentally exposed to goldfish. nfection rate was 22.9% out of 105 goldfish, and the encysted metacercariae were found in fins(86.7%) and on scales (13.7%) of the fishes, but not in their muscle, head or visceral organs. 3. Seven species of ask were caught in the Daecheong Reservoir and the upper stream. Infestations with metacercaria of Metagonimus were found 100% in Opsariichtys widens and the parasitized numbers of the metacercariae were observed from 250 to 2,400 per fish. In the upper stream, Zacco termmincki, Z. platypus and Pseudogobio esocinus were infected 100% with the metacercaria, on the other hand, the fishes caught in the reservoir showed the lower infestation rates, and a few metacercariae found in the fishes Carassius carassius and Cyprinus carpio in the reservoir and the stream. The majority of metacercariae was detected only on the scales of fishes. 4. In order to know the infectivity and the distribution patterns in the intestine of hosts, rats and dogs were infected with the metacercariae obtained from O. bidens and Z. platypus. In addition the metacercariae obtained from Z. temmincki, P. esocinus and goldfish were given to the rats. The recovery rates of the worms in the small intestine of dogs were higher (63.3~65.8%) than those of the rats (3.5~31.6%). The flukes were found mostly in the middle and the lower part of small intestine of the rats and the dogs, but no worm was collected in the upper part of the intestine of rats. 5. The sixte of adult flukes varied by the hosts. In the adult cukes, oral sucker was smaller than ventral sucker, and the right and left testes were located diagonally, the uterine tubules circled around the upper left testis. The average egg sixte was $29.1{\times}1.7{\mu\textrm{m}}$. According to the above results, the cukes belonging to genus Metagonimus distributed along the Geum River was concluded to be identical with Miyata type of M. yokogawai as that Saito had proposed.

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Construction and In vitro Study of a Prx 6/Luc Vector System for Screening Antioxidant Compounds in the Transgenic Mice (항산화반응을 유발하는 물질의 검색에 적용할 수 있는 형질전환 마우스 생산을 위한 새로운 Prx 6/Luc 벡터시스템의 제조 및 폐암세포주에서 반응성 확인)

  • Lee, Young Ju;Nam, So Hee;Kim, Ji Eun;Hwang, In Sik;Lee, Hye Ryun;Choi, Sun Il;Kwak, Moon Hwa;Lee, Jae Ho;Jung, Young Jin;An, Beum Soo;Hwang, Dae Youn
    • Journal of Life Science
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    • v.23 no.2
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    • pp.167-174
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    • 2013
  • Peroxiredoxin 6 (Prx 6) is a member of the thiol-specific antioxidant protein family, which may play a role in protection against oxidative stress and in regulating phospholipid turnover. The aim of this study was to determine whether a human Prx 6/Luc vector was stably expressed and responded to antioxidants in a lung cell line (NCI-H460). To achieve this, the luciferase signal, hPrx 6 mRNA expression, and superoxide dismutase (SOD) activity were measured in transfectants with a hPrx 6/Luc plasmid after treatment with four antioxidant extracts, including Korea white ginseng (KWG), Korea red ginseng (KRG), Liriope platyphylla (LP), and red Liriope platyphylla (RLP). First, the hPrx 6/Luc plasmid was successfully constructed with DNA fragments of human Prx 6 promoter, amplified by PCR using genomic DNA isolated from NCI-H460 cells, and cloned into the pTransLucent reporter vector. The orientation and sequencing of the hPrx 6/Luc plasmid were identified with restriction enzyme and automatic sequencing. A luciferase assay revealed significant enhancement of luciferase activity in the four treatment groups compared with a vehicle-treated group, although the ratio of the increase was different within each group. The KRG- and LP-treated groups showed higher activity than the KWG- and RLP-treated groups. Furthermore, the luciferase activity against RLP occurred roughly in a dose-dependent manner. However, the level of endogenous hPrx 6 mRNA did not change in any group treated with the four extracts. The SOD activity was in agreement with the luciferase activity. Therefore, these results indicate that the hPrx 6/Luc vector system may successfully express and respond to antioxidant compounds in NCI-H460 cells. The data also suggest that the Prx 6/Luc vector system may be effectively applied in screening the response of hPrx 6 to antioxidant compounds in transgenic mice.

Forest Vegetation Structure in Maruguem (the Ridge Line) Area of Gitdaebaegibong to Jukryeong, Baekdudaegan (백두대간(깃대배기봉-죽령 구간) 마루금 주변의 산림식생구조)

  • Song, Ju Hyeon;Yun, Chung Weon
    • Journal of Korean Society of Forest Science
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    • v.108 no.2
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    • pp.147-167
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    • 2019
  • This study was conducted to analyze forest vegetation structure in the Marugeum (Ridge) area of Gitdaebaegibong to Jukryeong, Baekdudaegan. Data were collected in 298 quadrates through a Braun-Blanquet vegetation survey from April, 2018 to October, 2018. Forest vegetation was classified into 13 vegetation units. A Quercus mongolica community was divided into Morus bombycis, Filipendula glaberrima, Fraxinus sieboldiana, Prunus maackii unit and Q. mongolica typical unit. The M. bombycis unit was further classified into a Deutzia glabrata group and M. bombycis typical group. The F. glaberrima unit was subdivided into a Veratrum oxysepalum group, Arundinella hirta group, and F. glaberrima typical group. The F. sieboldiana unit was divided into a Pinus densiflora group, Larix kaempferi group, and F. sieboliana typical group. The relationship between vegetation units and environmental factors was studied through coincidence analysis and CCA. The F. glaberrima unit (VU 6~8) was distributed by elevation above 1,200 m and other vegetation units were distributed below 1,200 m. Results of the CCA analysis showed that the F. glaberrima unit distribution is positively correlated with elevation. As a result of species diversity, the F. glaberrima unit was higher than other vegetation units. A similarity index analysis revealed that the F. sieboldiana unit (VU 9~11) was relatively homogeneous, and the M. bombycis unit (VU 1~5) and A. girta group (VU 7) were relatively heterogeneous. A detrended correspondence analysis determined that the distance between the statistical axes of the M. bombycis and F. glaberrima units was the greatest, which is consistent with the analysis of the similarity index. As a result of interspecific correlation of major woody plants, hydrophilic species were positively correlated, and a negative correlation was found between Q. mongolica and intolerant species such as P. densiflora and L. kaempferi.

Antioxidative Effects of Tenebrio molitor Larvae Extract Against Oxidative Stress in ARPE-19 Cells (ARPE-19 세포에서 산화적 스트레스에 대한 갈색거저리 추출물의 항산화 효과)

  • Bong Sun, Kim;Ra-Yeong, Choi;Eu-Jin, Ban;Joon Ha, Lee;In-Woo, Kim;Minchul, Seo
    • Journal of Life Science
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    • v.32 no.11
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    • pp.865-871
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    • 2022
  • Tenebrio molitor larvae is well known as edible insect. Then, although it has been widely studied that Tenebrio molitor larvae has various bioactive functions such as antioxidant, anti-wrinkle, and anticancer. Nevertheless, antioxidant effects of Tenebrio molitor larvae water extract (TMH) has not been well described in Adult Retina Pigment Epithelial cell line (ARPE-19). In this study, we demonstrated that antioxidant effects of TMH against H2O2-induced oxidative stress in ARPE-19. Thus, we selected for our studies and performed a series of dose-response assay to determine the working concentration that lead to a consistent and high degree of cytotoxicity, which we defined as the level of H2O2 that killed 40% of the ARPE-19 cells. ARPE-19 cells were pre-treated with various concentrations of TMH (0.1 up to 2 mg/ml) before exposure to 300 µM H2O2. As we expected, TMH effectively prevented ARPE-19 cells from 300 µM H2O2-induced cell death in a dose-dependent manner. Furthermore, TMH inhibited the phosphorylation of mitogen-activated protein kinases (MAPKs) such as extracellular signal regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. Overall, the inhibitory effects of TMH on H2O2-induced apoptosis and oxidative stress were associated with the protection cleaved caspase-3, Bax, Bcl-2, and HO-1. The TMH suppressed H2O2-induced cell membrane leakage and oxidative stress in ARPE-19 cells. Thus, these results suggest that the TMH plays an important role in antioxidant effect in ARPE-19.

Project of Improving Good Agriculture Practice and Income by Intergrated Agricultural Farming (미얀마 우수농산물 재배기술 전수사업)

  • Lee, Young-Cheul;Choi, Dong-Yong
    • Journal of Practical Agriculture & Fisheries Research
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    • v.16 no.1
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    • pp.193-206
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    • 2014
  • The objectives of the project are to increase farmers' income through GAP and to reduce the loss of agricultural produce, for which the Korean partner takes a role of transferring needed technologies to the project site. To accomplish the project plan, it is set to implement the project with six components: construction of buildings, installation of agricultural facilities, establishment of demonstration farms, dispatching experts, conducting training program in Korea and provision of equipments. The Project Management Committee and the Project Implementation Team are consisted of Korean experts and senior officials from Department of Agriculture, Myanmar that managed the project systematically to ensure the success of the project. The process of the project are; the ceremony of laying the foundation and commencing the construction of training center in April, 2012. The Ribbon Cutting Ceremony for the completion of GAP Training Center was successfully held under PMC (MOAI, GAPI/ARDC) arrangement in SAl, Naypyitaw on June 17, 2012. The Chairman of GAPI, Dr. Sang Mu Lee, Director General U Kyaw Win of DOA, officials and staff members from Korea and Myanmar, teachers and students from SAl attended the ceremony. The team carried out an inspection and fixing donors' plates on donated project machineries, agro-equipments, vehicles, computers and printer, furniture, tools and so forth. Demonstration farm for paddy rice, fruits and vegetables was laid out in April, 2012. Twenty nine Korean rice varieties and many Korean vegetable varieties were introduced into GAP Project farm to check the suitability of the varieties under Myanmar growing conditions. Paddy was cultivated three times in DAR and twice in SAl. In June 2012, vinyl houses were started to be constructed for raising seedlings and finished in December 2012. Fruit orchard for mango, longan and dragon fruit was established in June, 2012. Vegetables were grown until successful harvest and the harvested produce was used for panel testing and distribution in January 2013. Machineries for postharvest handling systems were imported in November 2012. Setting the washing line for vegetables were finished and the system as run for testing in June 2013. New water tanks, pine lines, pump house and electricity were set up in October 2013.

Estimation of the CY Area Required for Each Container Handling System in Mokpo New Port (목표 신항만의 터미널 운영시스템에 따른 CY 소요면적 산정에 관한 연구)

  • Keum, J.S.
    • Journal of Korean Port Research
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    • v.12 no.1
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    • pp.35-46
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    • 1998
  • The CY can be said to function in various respect as a buffer zone between the maritime and overland inflow-outflow of container. The amount of storage area needed requires a very critical appraisal at pre-operational stage. A container terminal should be designed to handle and store containers in the most efficient and economic way possible. In order to achieve this aim it is necessary to figure out or forecast numbers and types of containers to be handled, CY area required, and internal handling systems to be adopted. This paper aims to calculate the CY area required for each container handling system in Mokpo New Port. The CY area required are directly dependent on the equipment being used and the storage demand. And also the CY area required depends on the dwell time. Furthermore, containers need to be segregated by destination, weight, class, FCL(full container load), LCL(less than container load), direction of travel, and sometimes by type and often by shipping line or service. Thus the full use of a storage area is not always possible as major unbalances and fluctuations in these flow occuring all the time. The calculating CY area must therefore be taken into account in terms of these operational factors. For solving such problem, all these factors have been applied to estimation of CY area in Mokpo New Port. The CY area required in Mokpo New Port was summarized in the conclusion section.

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A Study of the Influencing Factors for Decision Making on Construction Contract Types : Focused on DoD Construction Acquisitions with Firm Fixed Price and Cost Reimbursable in FAR (건설공사 대가지급방식의 의사결정 영향요인에 관한 연구 - 미국 연방조달규정에 따른 미국 국방성의 정액계약과 실비정산계약을 중심으로 -)

  • Son, Young-Hoon;Kim, Kyung-Rai
    • Korean Journal of Construction Engineering and Management
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    • v.25 no.2
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    • pp.23-35
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    • 2024
  • This study analyzed the correlation between each of the 12 influencing factors in FAR 16.04 and the decision-making process for construction contract types, using data from a total of 2,406 DoD Construction Acquisitions spanning from 2008 to 2022. The study considered 12 independent variables, grouped into 4 Characteristics with 3 factors each. Meanwhile, all other contract types were categorized into two types: Firm-Fixed-Price (FFP) and Cost-Reimbursement Contract (CRC), which served as the dependent variables. The findings revealed that FFP contracts significantly dominated in terms of acquisition volume. In line with prevailing beliefs, logistic data analysis and Analytical Hierarchy Process (AHP) analysis of Relative Weights from Experts' Survey demonstrated that independent variables like Uncertainty of the Scope of Work and Complexity found out to be increasing the likelihood of selecting CRC. The number of contractors in the market does indeed influence the possibilities of contract decision-making between CRC and FFP. Meanwhile, the p-values of the top 3 influencing factors on CRC from the AHP analysis-namely, Appropriateness of CAS, Project Urgency, and Cost Analysis-exceeded 0.05 in the binominal regression results, rendering it inconclusive whether they significantly influenced the construction contract type decision, particularly with respect to payment methods. This outcome partly results from the fact that a majority of respondents possessed specific experiences related to the USFK relocation project. Furthermore, influencing factors in construction projects behave differently than common beliefs suggest. As a result, it is imperative to consider the 12 influencing factors categorized into 4 Characteristics areas before establishing acquisition strategies for targeted construction projects.

Expression of TIMP1, TIMP2 Genes by Ionizing Radiation (이온화 방사선에 의한 TIMP1, TIMP2 유전자 발현 측정)

  • Park Kun-Koo;Jin Jung Sun;Park Ki Yong;Lee Yun Hee;Kim Sang Yoon;Noh Young Ju;Ahn Seung Do;Kim Jong Hoon;Choi Eun Kyung;Chang Hyesook
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.171-180
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    • 2001
  • Purpose : Expression of TIMP, intrinsic inhibitor of MMP, is regulated by signal transduction in response to genotoxins and is likely to be an important step in metastasis, angiogenesis and wound healing after ionizing radiation. Therefore, we studied radiation mediated TIMP expression and its mechanism in head and neck cancer cell lines. Materials and Methods : Human head and neck cancer cell lines established at Asan Medical Center were used and radiosensitivity $(D_0)$, radiation cytotoxicity and metastatic potential were measured by clonogenic assay, n assay and invasion assay, respectively. The conditioned medium was prepared at 24 hours and 48 hours after 2 Gy and 10 Gy irradiation and expression of TIMP protein was measured by Elisa assay with specific antibodies against human TIMP. hTIMP1 promoter region was cloned and TIMP1 luciferase reporter vector was constructed. The reporter vector was transfected to AMC-HN-1 and -HN-9 cells with or without expression vector Ras, then the cells were exposed to radiation or PMA, PKC activator. EMSA was peformed with oligonucleotide (-59/-53 element and SP1) of TIMP1 promoter. Results : $D_0$ of HN-1, -2, -3, -5 and -9 cell lines were 1.55 Gy, 1.8 Gy, 1.5 Gt, 1.55 Gy and 2.45 Gy respectively. n assay confirmed cell viability, over $94\%$ at 24hrs, 48hrs after 2 Gy irradiation and over 73% after 10 Gy irradiation. Elisa assay confirmed that cells secreted TIMP1, 2 proteins continuously. After 2 Gy irradiation, TIMP2 secretion was decreased at 24hrs in HN-1 and HN-9 cell lines but after 10 Gy irradiation, it was increased in all cell lines. At 48hrs after irradiation, it was increased in HN-1 but decreased in HN-9 cells. But the change in TIMP secretion by RT was mild. The transcription of TIMP1 gene in HN-1 was induced by PMA but in HN-9 cell lines, it was suppressed. Wild type Ras induced the TIMP-1 transcription by 20 fold and 4 fold in HN-1 and HN-9 respectively. The binding activity to -59/-53, AP1 motif was increased by RT, but not to SP1 motif in both cell lines. Conclusions : We observed the difference of expression and activity of TIMPs between radiosensitive and radioresistant cell line and the different signal transduction pathway between in these cell lines may contribute the different radiosensitivity. Further research to investigate the radiation response and its signal pathway of TIMPs is needed.

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