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Viabilities of Biopsied Mouse Embryos after Ultrarapid Refreezing and Thawing (미세조작된 생쥐수정란의 초급속 재동결융해 후 생존성)

  • 신상태;임준호;강만종;한용만;이경광
    • Korean Journal of Animal Reproduction
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    • v.20 no.2
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    • pp.207-214
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    • 1996
  • To examine the developmental capacity of manipulated embryos after ultrarapid refreezing and thawing, mouse embryos were biopsied at 4-cell stage, frozen twice at 4-cell and morula stages, respectively, and then transferred to rec-ipients. Single blastomeres were biopsied from 4-cell embryos by a modified aspiration method. Biopsied 4-cell embryos were equilibrated into freezing medium at room temperature for 2.5 min, loaded into 40 $\mu$I of freezing medium in 0.25 ml plastic straw and then directly immersed into liqiud nitrogen. Freezing medium for 4-cell embryos consisted of 4.0 M ethylene glycol and O.25 M sucrose in dPBS supplemented with 6 mg/lm BSA. Morulae were frozen into freezing medium containing 5.0 M glycerol instead of ethylene glycol. Thawing was conducted by agitating each straw in 3TC water for 20 sec. The c content of each straw was expelled into 0.5 ml of dilution medium, which consisted of 0.25 M sucrose and 3 mg/ml BSA in dPBS. The thawed embryos were rehydrated in dilution medium for 10 min, washed 3 times with dPBS and then cultured in M16 medium at 37$^{\circ}C$, 5% CO$_2$ in air. Blastocysts that developed from frozen or refrozne biopsied embryos were transferred to recipients on Day 3 of pseudopregnancy, respectively. In vitro and in vivo developmental rates of the biopsied and intact 4~cell embryos after freezing and thawing were 78 (10l/130) and 25% (10/40), and 91 (114/125) and 30% (12/40), respectively. Although the rates of in vitro development of biopsied and intact embryos to blastocyst stage were significantly different after freezing and thawing (P

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Cell Protective Effects of Enzymatic Hydrolysates of Citrus Peel Pectin (귤피 펙틴 유래 효소적 가수분해물의 세포 보호 효과)

  • Kwon, Soon Woo;Ko, Hyun Ju;Bae, Jun Tae;Kim, Jin Hwa;Lee, Geun Soo;Pyo, Hyeong Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.1
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    • pp.75-85
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    • 2016
  • Pectin, a naturally occurring polysaccharide, has in recent years attracted considerable attention. Its benefits are increasingly appreciated by scientists and consumers due to its safety and usefulness. The chemistry and gel-forming characteristics of pectin have enabled to be used in pharmaceutical industry, health promotion and treatment. Yet, it has been rarely used in cosmetics because of its incompatibility with many cosmetic ingredients, including alcohols, and unstable viscosity of pectin gels under various pH and salt conditions. However, low-molecular-weight pectin oligomers have excellent biological activities, and depolymerization of pectin to produce cosmetic ingredients would be very useful. In this study, we attempted the development of cosmetic ingredients using pectin with an excellent effect on human skin. We developed a bio-conversion process that uses enzymatic hydrolysis to produce pectin hydrolysates containing mainly low-molecular-weight pectin oligomers. Gel permeation chromatography was used to determined the ratio of hydrolysis. The molecular weight of the pectin hydrolysates obtained varied between 200 and 2,700 Da. The two newly developed low-molecular-weight pectin hydrolysates, LMPH A and B, had higher anti-oxidative activities than pectin or D-galacturonic. Exposure to UVB radiation induces apoptotic cell death in epidermal cells. Annexin V binding and propidium iodide uptake were measured by flow cytometry to evaluate UVB-induced cell death in HaCaT cells. Both LMPH A and B reduced UVB-induced cell death and increased cell proliferation by 22% and 30% at 0.5% concentration respectively, while pectin had no significant activity. In conclusion, this study suggests that the newly developed low-molecular-weight pectin hydrolysates can be used as safe and biologically active cosmetic ingredients.

Neural pathway innervating ductus Deferens of rats by pseudorabies virus and WGA-HRP (흰쥐에서 WGA-HRP와 pseudorabies virus를 이용한 정관의 신경로에 대한 연구)

  • Lee, Chang-Hyun;Chung, Ok-Bong;Ko, Byung-Moon;Lee, Bong-Hee;Kim, Soo-Myung;Kim, In-Shik;Yang, Hong-Hyun
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.11-24
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    • 2003
  • This experimental studies was to investigate the location of PNS and CNS labeled neurons following injection of 2% WGA-HRP and pseudorabies virus (PRY), Bartha strain, into the ductus deferens of rats. After survival times 4-5 days following injection of 2% WGA-HRP and PRV, the rats were perfused, and their brain, spinal cord, sympathetic ganglia and spinal ganglia were frozen sectioned ($30{\mu}m$). These sections were stained by HRP histochemical and PRY inummohistochemical staining methods, and observed with light microscope. The results were as follows ; 1. The location of sympathetic ganglia projecting to the ductus deferens were observed in pelvic ganglion, inferior mesenteric ganglion and L1-6 lwnbar sympathetic ganglia. 2. The location of spinal ganglia projecting to the ductus deferens were observed in T13-L6 spinal ganglia. 3. The PRY labeled neurons projecting to the ductus deferens were observed in lateral spinal nucleus, lamina I, II and X of cervical segments. In thoracic segments, PRY labeled neurons were observed in dorsomedial part of lamina I, II and III, and dorsolateral part of lamina IV and V. Densely labeled neurons were observed in intermediolateral nucleus. In first lumbar segment, labeled neurons were observed in intermediolateral nucleus and dorsal commisural nucleus. In sixth lumbar segment and sacral segments, dense labeled neurons were observed in sacral parasympathetic nuc., lamina IX and X. 4. In the medulla oblongata, PRV labeled neurons projecting to the ductus deferens were observed in the trigeminal spinal nuc., A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nuc., rostroventrolateral reticular nuc., area postrema, nuc. tractus solitarius, raphe obscurus nuc., raphe pallidus nuc., raphe magnus nuc., parapyramidal nuc., lateral reticular nuc., gigantocellular reticular nuc.. 5. In the pons, PRV labeled neurons projecting to the ductus deferens were ohserved in parabrachial nuc., Kolliker-Fuse nuc., locus cooruleus, subcooruleus nuc. and AS noradrenalin cells. 6. In midbrain, PRV labeled neurons projecting to the ductus deferens were observed in periaqueductal gray substance, substantia nigra and dorsal raphe nuc.. 7. In the diencephalon, PRV labeled neurons projecting to the ductus deferens were observed in paraventricular hypahalamic nuc., lateral hypothalamic nuc., retrochiasmatic nuc. and ventromedial hypothalamic nuc.. 8. In cerebrum, PRV labeled neurons projecting to the ductus deferens were observed in area 1 of parietal cortex. These results suggest that WGA-HRP labeled neurons of the spinal cord projecting to the rat ductus deferens might be the first-order neurons related to the viscero-somatic sensory and sympathetic postganglionic neurons, and PRV labeled neurons of the brain and spinal cord may be the second and third-order neurons response to the movement of smooth muscles in ductus deferens. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitaing the internal environment. These observations provide evidence for previously unknown projections from ductus deferens to spinal cord and brain which may be play an important neuroanatornical basic evidence in the regulation of ductus deferens function.

Treatment Results of Esophageal Carcinoma Treated by Radiation Therapy (식도암의 방사선치료 성적)

  • Kim Mi Sook;Yoo Seoung Yul;Cho Chul Koo;Yoo Hyung Jun;Yang Kwang Mo;Kang Jin Oh;Ji Young Hoon;Lee Dong Han;Ryoo Baek Yeol
    • Radiation Oncology Journal
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    • v.18 no.3
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    • pp.182-186
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    • 2000
  • Purpose : To determine treatment or이ecol for inoperable esophageal cancer patients, 껜e evaluated survival rate and prognostic factors. Materials and Methods : We evaluated esophageal cancer treated by curative or palliative am in KCCH from 1992 to 1996, retrospectively. Recurrent or underdose case below 40 Gy were excluded. The number of male and female were 35 and 5, respectively. Thirty-eight patients were squamous carcinoma and 2 patients were not biopsy proven. Ten patients were treated with radiation therapy and chemotherapy Median dose of radiation therapy was 59.4 Gy and the range was $40\~60$ Gy. Results : The median survival is 6.5 months and 1-year survival rate was $28.3\%$. Age, location, radiation dose and chemotherapy were not significant prognostic factors. Median survivals of patients with below stage III and over stage IVA were 7.6 and 6.2 months respectively, but it is not significant. Conclusions : The survival for esophageal cancer is very poor. For patients with curative aim, chemotherapy must be considered. For patients with palliative aim, short-term external beam radiation therapy and/or brachytherapy must be considered.

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A Development and Application of the Landscape Evaluation Model Based on the Biotope Classification (비오톱 유형분류를 기반으로 한 경관평가 모형개발 및 적용)

  • Park, Cheon-Jin;Ra, Jung-Hwa;Cho, Hyun-Ju;Kim, Jin-Hyo;Kwon, Oh-Sung
    • Journal of the Korean Institute of Landscape Architecture
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    • v.40 no.4
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    • pp.114-126
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    • 2012
  • The purpose of this study is to find ways of the view evaluation of biotope classification before development by selecting an area, which is as large as about $10.0km^2$ around Non Gong Up, Auk Po Myun, Dalsung Gun, Daugu where the large project has been planned, as a subject of this study. The results of this study are as follows. Because of the classification of biotope, there are 23 kinds of types that are subdivided into 140 types. Three surveys for selecting the assessment indicators were performed. The first survey analyzed the importance of 22 selected assessment indicators based on the evaluation of an existing literature review and on the spot research. The second survey performed factor analysis and reclassified the value indicators. The third survey computed additive values of the selected assessment indicators. It used a method of standardizing the average importance of indicators by making their sum equal by 10. Theses additive values were then multiplied by each grade of indicators in order to make a final evaluation. The number of assessment indicators finally selected through the survey of asking specialist is vitality elements, visual obstructs elements etc 19. According to the result of evaluation of 1st, 1 grade spaces which especially valuable is analyzed that 7 spaces, 2 grade spaces for 4, 3 grade spaces for 5, 4 grade space for 2, 5 grade space for 5. Because of the evaluation of 2st, 1 grade spaces which especially valuable(1a, 1b) is analyzed that 15 spaces, 2 grade spaces which valuable is analyzed that 28 space. As the evaluation of site suitability model of this study couldn't have high applicability to other similar area because of having only one site as a subject, it is needed to do synthesize and standardization of various examples to have higher objectivity later.

Standardization and Reliability and Validity of the Korean Edition of Profile of Mood States(K-POMS) (한국판 기분상태척도(K-POMS)의 표준화와 신뢰도와 타당도 평가)

  • Kim, Eui-Joong;Lee, Sang-Ick;Jeong, Do-Un;Shin, Min-Sup;Yoon, In-Young
    • Sleep Medicine and Psychophysiology
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    • v.10 no.1
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    • pp.39-51
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    • 2003
  • Objectives: The Profile of Mood States (POMS) has been used broadly and frequently in various medical studies with various subjects. For the development and application of the Korean edition of POMS, called K-POMS, we translated POMS into Korean and then evaluated the reliability and validity of K-POMS. Methods: The subjects were 161 male students with a mean age of 18.1 years (15-30) and 244 female students with a mean age of 16.6 years (14-23). They were requested to perform K-POMS and SCL-90-R (Korean version) at the same time. Means, reliability coefficients, and test-retest correlations of K-POMS were calculated. Content validities, correlations with SCL-90-R, and discrimination validities in comparison with 76 depressive patients were obtained, and factor analyses were carried out. Results: Mean scores of Total Mood Disturbance (TMD) and some subscales (T, F, C, V factor) showed a significant difference between male and female students. The internal consistency coefficient ($\alpha$) of the total 65 items was 0.93, and ranged from 0.67 to 0.90 for subscales. Test-retest correlations of 43 subjects ranged from 0.27 to 0.63. Seven psychiatrists rated the properness of the content of each item as more than moderate degree (mean scale points=2.66, in the range of 0 to 4). Every subscale of KPOMS exhibited significant correlation with the matching subscale of SCL-90-R. Mean scores of K-POMS of the total 405 subjects were significantly different from those of the 76 depressive subjects after age correction. Six factors were extracted, accounting for 51.2% of total variance. Factor 6 consisted of 6 items which came from various subscales of POMS and represented "uncertainty and helplessness," which may be somewhat different to depression or sadness proper. Conclusion: The validity and reliability of K-POMS could be confirmed. The significant difference in K-POMS scores between depressive subjects and normal subjects suggests that K-POMS can be used clinically for the evaluation and screening of depression. The main structure of K-POMS is very similar to that of POMS, except the Confusion-Bewilderment factor which shows weak factor consistency in K-POMS. This may reflect some emotional characteristic of Koreans such as an undifferentiated aspect of cognitive efficiency. The sixth factor newly extracted from K-POMS also may be indicative of an unique emotional aspect of young Koreans.

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A study on the smile of Korean Youth compared with twenty years ago (20년전과 비교한 한국청년의 미소 변화에 관한 연구)

  • Beom, Seung-Gyun;Jung, Ji-Hye;Kim, Tae-Young;Lee, Kyung-Eun;Dong, Jin-Keun
    • The Journal of Korean Academy of Prosthodontics
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    • v.50 no.3
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    • pp.162-168
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    • 2012
  • Purpose: Teeth are generally exposed when people smiling. Moreover, the exposed teeth and soft tissue when smiling becomes an important guideline for esthetically prosthetic restoration. This research is to compare and find out differences of Korean young men's smile living in presence and twenty years ago. Materials and methods: Subjects, 100 young men (50 male and 50 female), were required about several aspects; normally developed physical condition, no psychological or genetic disorders, a fine face with no loss of teeth, no experience in orthodontic or prosthetic treatment, relatively normal occlusion, aged between 20-29. The photos of the subject at rest position and front face when fully smiled were taken three times. 100 photos (50 male and 50 female) were chosen at random from the 240 university students' smile photos taken by Yoon and his colleagues in 1991. By Hulsey's method of measuring smile, several factors; the change of upper lip curvature, the change of the relation between the upper lip and teeth, parallelism between Mx. incisor and lower lip, contact relation between Mx. incisor and lower lip and teeth displayed in a smile, were measured and analyzed. Meanwhile, ten dentists assessed aesthetic evaluation about men and women's smile for twice and recorded and compared smile score. The -test (P<.05) was used to compare the measured value. The difference of smile score was analyzed by t-test (P<.05). Results: The smile score calculated in 2011 (60.22) was higher than that of 1991 (52.80). Among five measurement categories, the noticeable difference was distinguished from two factors; the change of upper lip curvature and contact relation between Mx. incisor and lower lip. Conclusion: The Korean young men's smile has been considerably improved for twenty years. And it is found that the change of upper lip curvature plays an important role, that is, the smile formed with an ascended labial commissure has been increased significantly.

INVESTIGATION OF THE EFFECT OF AN ANTIBIOTIC "P" ON POTATOES ("감자에 대한 항생제(抗生劑) 피마리신의 통계적(統計的) 효과(效果) 분석(分析)")

  • Kim, Jong-Hoon
    • Journal of Korean Society for Quality Management
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    • v.5 no.2
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    • pp.59-120
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    • 1977
  • An antibiotic 'P', which is one of the products of the Gist Brocades N. V. is being tested by its research department as fungicide on seed-potatoes. For this testing they designed experiments, with two control groups, one competitor's product, eight formulations of the antibiotic to be tested in different concentrations and one mercury treatment which can not be used in practice. The treated potatoes were planted in three different regions, where bifferent conditions prevail. After several months the harvested potatoes are divided in groups according to their diameter, potato illness is analysed and counted. These data were summarised in percentage and given to us for Analysis. We approached and analysed the data by following methods: a. Computation of the mean and standard deviation of the percenage of good results in each size group and treatment. b. Computation of the experimental errors by substraction of each treatment mean from observed data. c. Description of the frequency table, plotting of a histogram and a normal curve on same graph to check normality. d. Test of normality paper and chi-sqeare test to check the goodness of fit to a normal curve. e. Test for homogeneity of variance in each treatment with the Cochran's test and Hartley's test. f. Analysis of Variance for testing the means by one way classifications. g. Drawing of graphs with upper and lower confidence limits to show the effect of different treatments. h. T-test and F-test to two Control mean and variance for making one control of Dunnett's test. i. Dunnett's Test and calculations for numerical comarision of different treatments wth one control. In region R, where the potatoes were planted, it was this year very dry and rather bad conditions to grow potatoes prevailed during the experimental period. The results of this investigation show us that treatment No.2, 3 and 4 are significantly different from other treatments and control groups (none treated, just like natural state). Treatment no.2 is the useless mercury formulation. So only No. 3 and 4, which have high concentrations of antibiotic 'P', gave a good effect to the potatoes. As well as the competitors product, middle and low concentrated formulations are not significantly different from control gro-ups of every size. In region w, where the potatoes got the same treatments as in region R, prevailed better weather conditions and was enough water obtainable from the lake. The results in this region showed that treatment No. 2, 3, 4, and 5 are Significantly different from other treatments and the control groups. Again No.2 is the mercury treatmentin this investigation. Not only high concentrated formulation of antibiotic 'P', but also the competitor's poroduct gave good results. But, the effect of 'P', was better than the competitors porduct. In region G, where the potatoes got the same treatments as in the regions R and w. and the climate conditions were equal to region R, the results showed that most of the treatments are not significantly different from the control groups. Only treatment no. 3 was a little bit different from the others. but not Significantly different. It seems to us that the difference between the results in the three regions was caused by certain conditions like, the nature of the soil the degres of moisture and hours of sunshine, but we are not sure of that. As a conclusion, we can say that antibiotic 'P' has a good effect on potatoes, but in most investigations a rather high concentration of 'P' was required in formulations.

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Comparison of Inhalation Scan and Perfusion Scan for the Prediction of Postoperative Pulmonary Function (수술후 폐기능 변화의 예측에 대한 연무 흡입스캔과 관류스캔의 비교)

  • Cheon, Young-Kug;Kwak, Young-Im;Yun, Jong-Gil;Zo, Jae-Ill;Shim, Young-Mog;Lim, Sang-Moo;Hong, Sung-Woon;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.2
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    • pp.111-119
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    • 1994
  • Background: Because of the common etiologic factor, such as smoking, lung cancer and chronic obstructive pulmonary disease are often present in the same patient. The preoperative prediction of remaining pulmonary function after the resectional surgery is very important to prevent serious complication and postoperative respiratory failure. $^{99m}Tc$-MAA perfusion scan has been used for the prediction of postoperative pulmonary function, but it may be inaccurate in case of large V/Q mismatching. We compared $^{99m}Tc$-DTPA radioaerosol inhalation scan with $^{99m}Tc$-MAA perfusion scan in predicting postoperative lung function. Method: Preoperative inhalation scan and/or perfusion scan were performed and pulmonary function test were performed preoperatively and 2 month after operation. We predicted the postoperative pulmonary functions using the following equations. Postpneurnonectomy $FEV_1$=Preop $FEV_1x%$ of total function of lung to remain Postlobectomy $FEV_1$=Preop $FEV_1{\times}$(% of total 1-function of affected lung${\times}$$\frac{Number\;of\;segments\;to\;be\;resected}{Number\;of\;segments\;of\;affected\;lung})$ Results: 1) The inhalation scan showed good correlations between measured and predicted $FEV_1$, FVC and $FEF_{25-75%}$. (correlation coefficiency; 0.94, 0.91, 0.87 respectively). 2) The perfusion scan also showed good correlations between measured and predicted $FEV_1$, FVC and $FEF_{25-75%}$. (correlation coefficiency; 0.86, 0.72, 0.87 respectively). 3) Among three parameters, $FEV_1$ showed the best correlations in the prediction by lung scans. 4) Comparison between inhalation scan and perfusion scan in predicting pulmonary function did not show any significant differneces except FVC. Conclusion: The inhalation scan and perfusion scan are very useful in the prediction of postoperative lung function and don't make a difference in the prediction of pulmonary function a1though the former showed a better correlation in FVC.

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Effects of Matrix Metalloproteinase Inhibitor on Ventilator-Induced Lung Injury in Rats (기계환기로 인한 백서의 급성 폐손상에서 Matrix Metalloproteinase Inhibitor의 효과)

  • Kim, Je-Hyeong;Park, Soo-Yeon;Hur, Gyu-Young;Lee, Seung-Heon;Lee, Sang-Yeub;Park, Sang-Myeon;Suh, In-Bum;Shin, Chol;Shim, Jae-Jeong;In, Kwang-Ho;Kang, Kyung-Ho;Yoo, Se-Hwa
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.6
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    • pp.619-634
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    • 2002
  • Background : Many inflammatory mediators and collagenases are involved in the pathogenesis of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). The increase of matrix metalloproteinase-9 (MMP-9, gelatinase-B) produced mainly by inflammatory cells was reported in many ALI models and connective tissue cells. In this study, the expression of MMP-9 in ventilator-induced lung injury (VILI) model and the effects of matrix metalloproteinase inhibitor (MMPI) on VILI were investigated. Methods : Eighteen Sprague-Dawley rats were divided into three groups: low tidal Volume (LVT, 7mL/Kg tidal volume, 3 $cmH_2O$ PEEP, 40/min), high tidal volume (HVT, 30mL/Kg tidal volume, no PEEP, 40/min) and high tidal volume with MMPI (HVT+MMPI) groups. Mechanical ventilation was performed in room air for 2 hours. The 20 mg/Kg of CMT-3 (chemically modified tetracycline-3, 6-demethyl 6-deoxy 4-dedimethylamino tetracycline) was gavaged as MMPI from three days before mechanical ventilation. The degree of lung injury was measured with wet-to-dry weight ratio and acute lung injury score. Expression of MMP-9 was studied by immunohistochemical stain with a mouse monoclonal anti-rat MMP-9 $IgG_1$. Results : In the LVT, HVT and HVT+MMPI groups, the wet-to-dry weight ratio was $4.70{\pm}0.14$, $6.82{\pm}1.28$ and $4.92{\pm}0.98$, respectively. In the HVT group, the ratio was significantly higher than other groups (p<0.05). Acute lung injury score measured by five-point scale was $3.25{\pm}1.17$, $12.83{\pm}1.17$ and $4.67{\pm}0.52$, respectively. The HVT group was significantly damaged by VILI and MMPI protects injuries by mechanical ventilation (p<0.05). Expression of MMP-9 measured by four-point scale was $3.33{\pm}2.07$, $12.17{\pm}2.79$ and $3.60{\pm}1.95$, respectively, which were significantly higher in the HVT group (p<0.05). Conclusion : VILI increases significantly the expression of MMP-9 and MMPI prevents lung injury induced by mechanical ventilation through the inhibition of MMP-9.