• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.681-692
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• 1996

The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

• Biomolecules & Therapeutics
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• v.20 no.1
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• pp.43-49
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• 2012

Stimulation of mast cells through the high affinity IgE receptor (Fc${\varepsilon}$RI) induces degranulation, lipid mediator release, and cytokine secretion leading to allergic reactions. Although various signaling pathways have been characterized to be involved in the Fc${\varepsilon}$RI-mediated responses, little is known about the precious mechanism for the expression of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in mast cells. Here, we report that rapamycin, a specific inhibitor of mammalian target of rapamycin (mTOR), reduces the expression of TNF-${\alpha}$ in rat basophilic leukemia (RBL-2H3) cells. IgE or specific antigen stimulation of RBL-2H3 cells increases the expression of TNF-${\alpha}$ and activates various signaling molecules including S6K1, Akt and p38 MAPK. Rapamycin specifically inhibits antigeninduced TNF-${\alpha}$ mRNA level, while other kinase inhibitors have no effect on TNF-${\alpha}$ mRNA level. These data indicate that mTOR signaling pathway is the main regulation mechanism for antigen-induced TNF-${\alpha}$ expression. TNF-${\alpha}$ mRNA stability analysis using reporter construct containing TNF-${\alpha}$ adenylate/uridylate-rich elements (AREs) shows that rapamycin destabilizes TNF-${\alpha}$ mRNA via regulating the AU-rich element of TNF-${\alpha}$ mRNA. The antigen-induced activation of S6K1 is inhibited by specific kinase inhibitors including mTOR, PI3K, PKC and $Ca^{2+}$chelator inhibitor, while TNF-${\alpha}$ mRNA level is reduced only by rapamycin treatment. These data suggest that the effects of rapamycin on the expression of TNF-${\alpha}$ mRNA are not mediated by S6K1 but regulated by mTOR. Taken together, our results reveal that mTOR signaling pathway is a novel regulation mechanism for antigen-induced TNF-${\alpha}$ expression in RBL-2H3 cells.

• Korean Journal of Weed Science
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• v.15 no.1
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• pp.30-38
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• 1995

Soil-applied pre-emergence herbicide, pyrazolate(4-(2, 4-dichlorobenzoyl)-1, 3-dimethyl pyrazol-5-yl-p-toluene sulphonate) induced, twist effect of shoots of barnyardgrass under dry conditions, and etiolated leaf and stem of that under water condition. Plant height and root length of rice broadcast on soil surface were similar to the untreated control, but plant height of rice drilled in soil was more inhibited than root length as compared with the untreated control, while development of barnyardgrass seedling was severely inhibited at 20 days after application. The inhibition rate was much higher under water condition than under dry condition, but difference in rice and barnyardgrass did not abserve. However, growth of transplanted rice shown to increase to the untreated control. Shoot and root fresh weight of rice broadcast on soil surface was increase as compared with the untreated control, and that of rice drilled in soil was not affected whereas that of barnyardgrass was severely inhibited by 42% and 41%, respectively. Under dry condition at 20 days after pyrazolate application while root growth of rice broadcast on soil surface under water condition was deadly inhibited and development of barnyardgrass was almost completely inhibited. On the other hand, microscopic studies showed that constriction of mesophyll cell by destruction of chloroplast of barnyardgrass were occurred only under dry condition, whereas damage of rice and barnyardgrass under water and transplanting condition were not observed. Anatomical change in the meristernatic region of rice and barnyardgrass was not occurred, and similar to intact plant regardless of cropping patterns.

• The Korean Journal of Nuclear Medicine
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• v.28 no.3
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• pp.317-325
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• 1994

The accuracy of dipyridamole stress/rest $^{99m}Tc$-MIBI myocardial imaging for detection of ischemia depends on reproducible image interpretation. To evaluate the reproducibility of visual assessment, agreement in interpretation among two independent observers, blind-ed to clinical data, was evaluated in SPECT images of 131 patients (94 males, 38 females; mean age $58{\pm}7yr$) with suspected coronary artery disease who underwent both dipyridamole stress/rest $^{99m}Tc$-MIBI myocardial SPECT and coronary angiography. The left ventricle was divided into twenty-nine segments in stress and rest SPECT images and each segment was visually graded according to a five-point scale (segmental score : 0=normal, 1=equivocal, 2=mild decrease, 3=severe decrease and 4=absent uptake). Overall concordance of segmental scoring between the two observers was 80%. The Pear-son's correlation coefficient (r) of the segmental scores for stress and rest images were 0.67 and 0.65, respectively, while the difference in score between the two images showed a correlation of 0.45 (all p<0.001). Agreement between two observers in final SPECT diagnosis as absence or presence of disease was 93%. The degree of agreement in segmental scoring showed no difference between patients with or without agreement as to the presence of disease. Therefore it appeared that cases with inconcordant diagnosis between the 2 observers were mainly due to a difference in individual threshold for interpretating the significance of a particular decreased uptake area rather than to a difference in perceiving the degree of the hypoactivity Thus, establishment of individual optimum thresholds in visual interpretation of myocardial SPECT may be helpful to improve reproducibility and accuracy of scan diagnosis.

• Korean Journal of Environmental Agriculture
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• v.19 no.1
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• pp.13-19
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• 2000

Seasonal variations of dominant algae species and the effects of these algae on coagulation and filteration of water treatment were investigated at Chilseo water treatment plant in downstream of Nakdong river from January in 1995 to Desember of 1998. The water quality of Nakdong river was found to be a hyper eutrophic state during the investigation periods. In the measurement, Chlorophyll-a contents ranged $20.7{\sim}180.9{\mu}g/l$ and total nitrogen contents(T-N) and total phosphorus contents(T-P) exceeded more than 3.4mg/l and 0.1mg/l, respectively. The changes in dominant algae species was in the order of Stepanodiscus sp., Asterionella sp., Melosira sp., Microcystis sp. and Synedra sp. from spring to winter. Microcystis sp. especially, was blooming during summer and Synedra sp. and Stepanodiscus sp. during winter. Although most diatomous algae appeared in the water treatment process caused filter clogging and reduced efficiency of coagulation and sedimentation, Synedra sp. and Stepanodiscus sp were revealed as the main trouble algae. Malfunction of water treatment process caused by Synedra sp. and Stepanodiscus sp. started at the algae concentrations of 800cells/ml and 1,820cells/ml, respectively. When chlorophyll-a content was $18.9{\mu}g/l$, the optimum amounts of coagulant were found to be 40mg/l of Alum and 16mg/l of PACS. Under condition of chlorophyll-a content of $154.1{\mu}g/l$, addition of Alum at the level of 75mg/l and PACS at the level of 35mg/l showed the lowest turibidity. The result indicates that increased amounts of the coagulants should be added for a better water treatment as chlorophyll-a contents increased. Addition of Alum at the amount of 60mg/l and 30mg/l of PACS removed Stepanodiscus sp. algae at the rate of 85% and 83%, respectively. In case of Synedra sp., 50mg/l of Alum and 25mg/l of PACS showed removal rates of 79% and 81%, respectively. Synedra sp. algae at the standing crops of 1,500cells/ml started filter clogging and a filtering process was completely inhibited after 8 hours. At this situation the filter clogging by Synedra sp. algae occurred at the depth of 5cm from the top anthracite layer. On the other, other algae did filter clogging at the depth of 10cm.

• Clinics in Shoulder and Elbow
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• v.16 no.2
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• pp.84-93
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• 2013

Purpose: The purpose of this study was to evaluate the repair integrity and clinical outcomes of delaminated tears following arthroscopic layered suture bridge rotator cuff repair in a prospective fashion. Materials and Methods: A consecutive series of 67 patients with delaminated rotator cuff tears who underwent surgery using the arthroscopic Layered Suture Bridge Technique were followed prospectively. Of 67 shoulders, 26 patients were male and 40 patients were female (one patient had bilateral tears); the mean age was 58.8(40~76) years. The clinical evaluation was performed according to ASES, UCLA, and KSS (Korea Shoulder Scoring System) scores with an average follow up period of 33 months. Repair integrity was estimated using MRI, which was performed six months postoperatively. Results: The average clinical outcome in ASES, UCLA, and KSS scores showed significant improvement at the time of the final follow-up compared to preoperatively, from 50.2 to 92.3, 15 to 31.3, and 54.4 to 90.7, respectively (p<0.001 for all scores). Follow up by MRI showed that 16 shoulders had a type-I; 14, a type-II; five, a type-III; two, a type-IV; one, a type-V re-tear. The overall rate of re-tear (types IV and V) was 7.9%. Conclusion: Use of the Arthroscopic Layered Suture Bridge Technique can result in improved functional and structural outcome in cases of delaminated rotator cuff tear.

• Korean Journal of Animal Reproduction
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• v.9 no.1
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• pp.9-30
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• 1985

The study was carried out to find out the changes of the sex hormone levels in the milk of Holstein cows during the reproductive stages such as the estrous cycle, pregnancy and periparturient period. The FSH, LH, estradiol-17$\beta$ and progesterone from the milk samples were assayed by radioimmunoassay methods. The results of this study were summarized as follows: 1. The levels of progesterone and estradiol-17$\beta$ were similar among inter-quarters, but they were higher in after milking than before milking times, with no statistical significance. 2. The milk progesterone levels during the estrous cycles reached a peak mean level of 3.55$\pm$0.26ng/$m\ell$ at 15 days after estrus and they did not show any differences among the length of estrous cycles. The estradiol-17$\beta$ levels during the estrous cycles showed a peak level of 36.40$\pm$2.38pg/$m\ell$ at estrus, and decreased(17.20$\pm$0.46 pg/$m\ell$ to 18.65$\pm$1.26pg/$m\ell$) at luteal phase. 3. The FSH levels during the estrous cycles ranged from 2.25$\pm$0.23mIU/$m\ell$ to 4.35$\pm$0.24mIU/$m\ell$ showing significant changes. The LH levels during the estrous cycles gradually increased and remained a peak level of 10.90$\pm$0.36mIU/$m\ell$ from 20 to 25 days after estrus. 4. The progesterone levels during the pregnancy were decreased from 30 to 60 days after artificial insemination, and therafter continuously increased until 240 days. The estradiol-17$\beta$ levels during the pregnancy were 24.56$\pm$1.19pg/$m\ell$ at day 30 after artificial inseminaton, and increased rapidly until 180 days. The levles were agagin decreased by 26.17$\pm$3.03pg/$m\ell$ until 210 days and markedly increased by 68.00$\pm$8.70pg/$m\ell$ until 240 days. 5. The prolactin levels during the pregnancy were 31.27$\pm$2.31ng/$m\ell$ and 42.60$\pm$2.37ng/$m\ell$ at day 150 and 240 after artificial insemination respectively. The LH levels during the pregnancy reached a peak of 27.47$\pm$7.90mIU/$m\ell$ at day 30 after artificial insemination, and thereafter gradually decreased. 6. The progesterone levels during the periparturient period reached a peak of 4.61$\pm$0.34ng/$m\ell$ at day 3 prepartum, and thereafter gradually decreased, and showed 2.05$\pm$0.60ng/$m\ell$ at day 7 postpartum. The estradiol-17$\beta$ levels during the periparturient period showed high level from 207.23$\pm$6.04pg/$m\ell$ at day 1 prepartum to 239.90$\pm$13.90pg/$m\ell$ at day 2 prepartum, and thereafter began to decline and reached 51.87$\pm$1.72pg/$m\ell$ at by 7 postpartum. 7. The prolactin levels during the periparturient period showed relatively higher level at the time of parturition. The LH levels during the periparturient period rnage from 6.32$\pm$0.32mIU/$m\ell$ to 13.90$\pm$1.37mIU/$m\ell$ showing significant changes. 8. The progesterone levels(4.6$\pm$0.8ng/$m\ell$) of the pregnant cows were significantly higher than those (1.84$\pm$1.4ng/$m\ell$) of nonpregnant cows. The cows of artificial insemination from 61 to 90 days after parturition showed higher progesterone levels. 9. During 20 to 25 days after artificial insemination, the accuracy of pregnancy diagnosis from milk progesterone levels were 94.4% for nonpregnant cows(<2.3ng/$m\ell$), and 75.0% for pregnant cows( 3.2ng/$m\ell$). The average overall accuracy of pregnancy prediction for nonpregnant and pregnant cows 83.3% 10. The results obtained this study suggest that the understanding of the endocrinological mechanisms by means of milk hormone analysis during the estrous cycle, pregnancy and parturition would give the basic information needed for increasing efficiency of reproduction. This study would not only provide an accurate method of the early pregnancy diagnosis by milk progesterone levels but also contribute to the research of providing the method of detecting of FSH levels in milk, which was difficult in blood serum.

• Proceedings of the Korea Society of Environmental Biology Conference
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• 2005.12a
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• pp.38-45
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• 2005

The egg bags of Korean salamander(Hynobius leechii) were collected from farmlands in Gyeongsangnam-do area. The assumed breeding time, numerical variation of embryos in each egg bag, mortality and the rates of abnormalities were investigated. The toxicity of benomyl, the metabolite carbendazim and BIC which were frequently spread in agricultural area and caused spontaneous embryonic malformation was investigated. The assumed breeding time between the end of February and the end of March has the difference about a month because of a habitat and it takes about 2 or 3 weeks from laying eggs to hatching. The length of each egg bag and the number of embryos were very varied in each area. It is due to geographical variation. Among egg bags in total study area, only 406 of egg bags(17.70% of total egg bags) developed all of embryos to normal larvae, and 78.49% of total embryos were normally developed. The patterns of spontaneous embryonic malformation were 26 species from A to Z and the abnormal patterns in individual were 8 species and above. the geographical differences about the abnormal pattern were identified and 11 habitats categorized 4 groups. The most frequent abnormality in Gyeongsangnam-do area is the dysplasia of external gill. The caudal dysplasia, abdominal blister and dysplasia of fin were also frequently observed. Individuals showing severe external defect were histologically studied and they showed retinal hypo-pigmentation, thyroid carcinoma, somatic muscular dysplasia, degeneration of cephalic neuron and various organ dysplasia. Benomyl and carbendazim were treated by 10pM$^{\sim}$10uM and BIC was treated by 1$^{\sim}$40ppm to know the effect of toxicity about toxic substance of salamander. After benomyl was treated, a survival rate was sharply dropped from 2 to 8 days. $LC_{100}$ identified in $1{\mu}M$, $LC_{50}$ identified between 100nM and $1{\mu}M$. $EC_{50}$ was assumed between 10nM and 100nM. The prevalent external malformation was abdomen swelled abnormally and histo-pathological effects were abdomen, neural tube and lens hernia. This suggests that benomyl is the toxicitic substance which inhibits the development of digestive system and nervous system. The result of treated carbendazim was similar to that of the treated benomyl. The survival rate is sharply dropped between 2 and 6 days. $LC_{100}$ was identified $1{\mu}M$ and $LC_{50}$ was identified between 10nM and 100nM. This shows that cabendazim has stronger lethal toxicity than benomyl. Ventral blister, eye dysplasia and cephalic dysplasia in the individual of external malformation mean that cabendazim affected nervous system much more than benomyl. Because the toxicity of BIC affected less in the beginning but affected more in the near hatching period, the period causing toxicity is somewhat different. $LC_{100}$ identified near 40ppm and $LC_{50}$ identified near 25ppm. The external defect shows mainly ventral blister and histo-pathological results show intestinal deformities. This result suggests the BIC inhibited strongly the development of digestive system. These abnormal developments may be caused by antimitotic action, inhibition of tubulin complex, destruction of microtubule, inhibitions of neurulation and closing of neural fold, and by the inhibition of movement of neural crest cells of benomyl. These abnormal developments may be caused by the rupture of epithelium, the loss of microtubule, the reduction of spindle size, the inhibition of spindle assembly formation, the destruction of spindle poles of carbendazim. These abnormal developments may be caused cytotoxicity by inhibition of the synthesis of a number of macromolecules and similar reaction the inhibition of benomyl.

• Applied Microscopy
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• v.38 no.4
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• pp.293-306
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• 2008

The essential factor of acute respiratory distress syndrome (ARDS), an acute lung injury accompanied commonly by sepsis syndrome is accumulation of neutrophils in lung tissue. The study attempted to confirm whether a lung injury would be decreased with the anti-inflammatory effect of germanium by the treated germanium prior to the development of ARDS and whether nitric oxide influence in suppressing a lung injury. Test groups were divided in the following structure for experiment; CON that has been administered with sodium chloride to airway, LPS administered with endotoxin for 5 hours in the same amount and 5 hours of endotoxin administered Ge+LPS following 1 hours of pre-treated germanium. The result of a test using experimental animals, infilteration of neutrophils (p<0.001) in bronchoalveolar lavage fluid (BALF) was significantly decreased, the structure of lung tissue was preserved relatively well, and much neutrophils with distinct positive were observed on tunel staining which showed increase of apoptotic neutrophils in the pre-treated germanium group compare to the endotoxin administrated group. In observation of ultrastructural changes of cell in BALF, phagocytic alveolar macrophage was increased in alveolar space, the nucleus of most engulfed neutrophils were condensed, and some apoptosis neutrophils appears to be DNA fragmentation and effacement of cellular organelles were found in intercellular matrix in the pre-treated germanium group. However, the nitric oxide showed increase in all the groups excluding CON, and the nitric oxide effect such as degranulation diminishing of mast cells and apoptosis increase of neutrophils in the pre-treated group only. The situation appears that there was change in internal environment of the experimental animal by the pre-treated germanium before the nitric oxide is produced and the anti-inflammatory effect activated the pre-processed germanium by nitric oxide which activated following the change. Therefore, the nitric oxide created from macrophage in accordance with the pre-treated germanium appears to influence in alleviating a lung injury. Accordingly, acute lung injury is alleviated by the anti-inflammatory effect of germanium such as inhibition of neutrophils migration, induction of neutrophil apoptosis and increase of phagocytic function of phagocyte, and the nitric oxide produced from activated macrophage by germanium would influence in suppressing a lung injury.

• Tuberculosis and Respiratory Diseases
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• v.50 no.2
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• pp.196-204
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• 2001

Background : Bronchial asthma is characterized by a reversible airway obstruction, airway hyperresponsiveness, and eosinophilic airway inflammation. The bronchodilator response(BDR) after short acting beta agonist inhalation and PC20 with methacholine inhalation are frequently used for diagnosing bronchial asthma. However, the relationship between the presence of a bronchodilator response and the degree of airway hyperresponsiveness is uncertain. Therefore, the availability of a eosinophil cationic protein (ECP) and a correlation ECP with a bronchodilator response and airway hyperresponsiveness was investigated. Method : A total 71 patients with a moderate to severe degree of bronchial asthma were enrolled and divided into two groups. 31 patients with a positive bronchodilator response and 38 patients with a negative bronchodilator response were evaluated. In both groups, the serum ECP, peripheral blood eosinophil counts, and total IgE level were measured and the methacholine bronchial provocation test was examined. Results : There were no differences observed in age, sex, atopy, and baseline spirometry in both groups. The peripheral eosinophil counts showed no difference in both groups, but the ECP level in group 1 (bronchodilator responder group) was higher than in group 2(non-bronchodilator responder group) ($22.4{\pm}20.7$ vs $14.2{\pm}10.4$, mean$\pm$SD). The PC20 in group 1 was significantly lower than in group 2 ($1.14{\pm}1.68$ vs $66{\pm}2.98$). There was a significant positive correlation between the BDR and ECP, and a negative correlation between the bronchial hyperresponsiveness and ECP. Conclusion : The bronchodilator response significantly correlated with the bronchial hyperresponsiveness and serum ECP in the moderate to severe asthma patients. Hence, the positive bronchodilator response is probably related with active bronchial inflammation and may be used as a valuable index in treatment, course and prognosis of bronchial asthma.