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Effects of $TGF-{\beta}1$ on Cellular Activity of Minocycline-Pretreated Human Periodontal Ligament Cells (($TGF-{\beta}$)이 Minocycline을 전처리한 사람 치주인대세포의 활성에 미치는 영향)

  • Yang, Seung-Oh;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • 제26권2호
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    • pp.469-490
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    • 1996
  • The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

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Evaluation of the accuracy of two different surgical guides in dental implantology: stereolithography fabricated vs. positioning device fabricated surgical guides (제작방법에 따른 임플란트 수술 가이드의 정확성비교: stereolithography와 positioning device로 제작한 수술 가이드)

  • Kwon, Chang-Ryeol;Choi, Byung-Ho;Jeong, Seung-Mi;Joo, Sang-Dong
    • The Journal of Korean Academy of Prosthodontics
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    • 제50권4호
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    • pp.271-278
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    • 2012
  • Purpose: Recently implant surgical guides were used for accurate and atraumatic operation. In this study, the accuracy of two different types of surgical guides, positioning device fabricated and stereolithography fabricated surgical guides, were evaluated in four different types of tooth loss models. Materials and methods: Surgical guides were fabricated with stereolithography and positioning device respectively. Implants were placed on 40 models using the two different types of surgical guides. The fitness of the surgical guides was evaluated by measuring the gap between the surgical guide and the model. The accuracy of surgical guide was evaluated on a pre- and post-surgical CT image fusion. Results: The gap between the surgical guide and the model was $1.4{\pm}0.3mm$ and $0.4{\pm}0.3mm$ for the stereolithography and positioning device surgical guide, respectively. The stereolithography showed mesiodistal angular deviation of $3.9{\pm}1.6^{\circ}$, buccolingual angular deviation of $2.7{\pm}1.5^{\circ}$ and vertical deviation of $1.9{\pm}0.9mm$, whereas the positioning device showed mesiodistal angular deviation of $0.7{\pm}0.3^{\circ}$, buccolingual angular deviation of $0.3{\pm}0.2^{\circ}$ and vertical deviation of $0.4{\pm}0.2mm$. The differences were statistically significant between the two groups (P<.05). Conclusion: The laboratory fabricated surgical guides using a positioning device allow implant placement more accurately than the stereolithography surgical guides in dental clinic.

Comparative Study on Eating Habits, Dietary Intake Patterns, and Nutrient Intakes Between Elementary School Children With and Without Atopic Dermatitis (일부 초등학생의 아토피피부염 유무에 따른 식습관, 식품섭취양상 및 영양소섭취상태 비교 연구)

  • Park, Nam-Suk;Jeon, Eu-Sun;Kim, Young-Nam;Cho, Kyung-Dong;Baek, Ok-Hee;Lee, Bog-Hieu
    • Journal of the Korean Society of Food Science and Nutrition
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    • 제38권11호
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    • pp.1543-1550
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    • 2009
  • The aim of this study was to examine and compare the eating habits, dietary intake patterns and nutrient intake status of children with and without atopic dermatitis by questionnaire during July, 2008. A total 388 subjects of 5th and 6th grade elementary school children (AG: atopic group, n=65, NG: non-atopic group, n=323) in Seoul and Ulsan areas participated in this study. The questionnaire included general characteristics, dietary habits, and atopy-related food frequency. One-day 24 hour recall was collected to estimate nutrient intakes of the subjects. The data were analyzed by Chi-square test for food frequency analysis and by t-test for nutrient intakes. Atopy-related foods included milk, buckwheat, beef, pork, chicken, crab, egg, mackerel, peach, and tomato. From the findings, AG had a more irregular eating habit than NG (p<0.05). In case of food frequency, AG tended to consume more atopy-related foods than NG (p>0.05). The nutrient intakes of AG were significantly lower than those of NG (p<0.05), but only intake of animal iron in AG was higher than NG (p<0.05). NG consumed more protein than AG (p<0.05). Although milk was a noted hypersensitive allergic food, frequency and the amount of milk intake were not significantly different between two groups. In conclusion, atopic children had eaten more atopy-related foods and less nutrient intakes. Therefore, it is necessary to educate on good nutrition and guide atopic children and their parents.

Quality Preservation of Shredded Carrots Stored in UV LED Packaging System (자외선 LED 포장용기 시스템에 의한 포장절단당근의 품질보존)

  • Kim, Nam Yong;Lee, Dong Sun;An, Duck Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • 제43권1호
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    • pp.135-140
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    • 2014
  • Pre-storage ultra-violet (UV) light treatment on fresh produce is known to inactivate the contaminated microorganisms, activate the defense system, and delay ripening extending the shelf life. As UV light emitting diode (LED) becomes available at a relatively low price, continuous or intermittent UV treatment during chilled storage is possible in a container or package. This study attempted an in situ UV LED treatment on fresh produce stored under a refrigerated container in order to see its potential in the fresh produce storage and further optimize its application conditions. The effect of in-container UV LED irradiation on the quality preservation of shredded carrots was investigated in the air and modified atmosphere (MA) conditions. Two sets of experiment with Escherichia coli inoculation and with natural microbial flora in the air (two 30 minute on-off cycles of 1 $diode/dm^2$ per day at a location above 2 cm) showed a clear and significant effect of the UV LED irradiation on the suppression of microbial growth: 280 nm was the most effective by maintaining a lower microbial count by at least 0.5 log (CFU/g) throughout the 6 day storage period. The carotenoids content of shredded carrots subjected to UV LED treatment at 365 and 405 nm in the air was higher than that of the control shredded carrots. In MA condition of $O_2$ of 1.2~4.3% and $CO_2$ of 8.4~10.6% being indifferent with LED wavelengths, 280 nm UV LED irradiation was also effective in inhibiting the microbial growth. While there was no observed difference in the carotenoids content between untreated and UV LED-treated shredded carrots in MA, UV LED irradiation at 365 and 405 nm was slightly better in DPPH radical scavenging activity. The use of UV LED in storage container or package seems to give the benefits of preserving the microbial and nutritional qualities of minimally processed fruits and vegetables.

Effects of Dietary Tea Polyphenol on Tumor Growth Inhibition by Cisplatin in EMT6 Breast Tumor-bearing Mice (유방암 세포(EMT6) 이식 마우스에서 녹차폴리페놀 음용이 시스플라틴의 암 조직 성장 억제에 미치는 영향)

  • Lee, Byoung-Rai;Cho, Jung-Il;Park, Pyoung-Sim
    • Journal of the Korean Society of Food Science and Nutrition
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    • 제43권1호
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    • pp.47-54
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    • 2014
  • The aim of this study is to evaluate the effects of green tea polyphenol (GTP) on anticancer treatment with cisplatin (CP), using both an in vitro cell culture model and an in vivo mouse model of established breast tumor. Mouse breast cancer cells (EMT6) were treated with or without GTP and CP followed by determination of the cell viability using an MTT assay. The relative cell viability of CP treated EMT6 cells was 96% at a 20 ${\mu}g/mL$ concentration of cisplatin; however, in combination with GTP (50 ${\mu}g/mL$), the cell viability decreased to 20% at the same concentration of CP (20 ${\mu}g/mL$). For the in vivo study, EMT6 cells were inoculated into Balb/c mice for the establishment of a tumor-bearing mice model. The tumor-bearing mice were treated with CP (5 mg/kg. i.p.) with or without dietary GTP (0.2% drinking water). Tumor growth was monitored by a measurement of tumor size using a digital caliper, and nephrotoxicity was determined by enzymatic and histological examinations. The levels of p53 and caspase-3 in tumor tissues were examined by a Western blot. In tumor-bearing mice treated with GTP plus CP, the increment of tumor volume showed a significant reduction, compared with CP or GTP alone. The levels of p53 and cleaved caspase-3 (caspase-3/p17) in tumor tissues of tumor-bearing mice were increased by CP and GTP compared to CP alone. In CP treated tumor-bearing mice, ${\gamma}$-glutamyltranspeptidase (GGT) and alkaline phosphatase (AP) activities were decreased, and marked tubular necrosis and dilatation were observed in the kidney. CP-induced enzymatic and histopathological changes in the kidney of tumor-bearing mice were reduced by combinations of GTP with CP. The results of these experiments demonstrated that dietary GTP has a potentiating effect on CP anti-tumor activity and a protective effect against CP-induced renal dysfunction. Therefore, GTP may be used as a modulator in anticancer treatment with CP.

Effects of Quality Characteristics and Antioxidant Activities of Yukwa Added with 'Donganme' Sorghum Bran Powder and Extracts (수수겨 분말 및 추출물 첨가가 유과의 품질특성 및 항산화활성에 미치는 영향)

  • Ko, Jee Yeon;Woo, Koan Sik;Kim, Jung In;Song, Seuk Bo;Lee, Jae Saeng;Jeong, Mi Seon;Jung, Tae Wook;Yun, Young Ho;Oh, In Seok
    • Journal of the Korean Society of Food Science and Nutrition
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    • 제43권8호
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    • pp.1181-1188
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    • 2014
  • This study investigated the quality characteristics and antioxidant activities of Yukwa added with 'Donganme' sorghum bran powder and its extracts. 'Donganme' variety has the highest antioxidant activity among sorghum varieties in Korea. The added contents of sorghum bran to Yukwa were 1% and 5% bran powder of 'Donganme' (BPD 1% and 5%) as well as 0.1%, 0.5%, and 1% bran powder extracts of 'Donganme' (BED 0.1%, 0.5%, and 1%). The contents of protein, ash, and minerals of BPD 1% and 5% added Yukwa were higher compared to non-added and BED 0.1%, 0.5%, and 1% added Yukwa. The contents of flavonoids of BPD 1% and 5% as well as BED 0.1%, 0.5%, and 1% added Yukwa increased by 1.6~17.1 fold, and DPPH and ABTS radical scavenging activities increased by 1.8~7.4 fold and 2.3~13.6 fold, respectively, compared to non-added Yukwa. Yukwa added with BED 1% showed the highest antioxidant activity among the treatments, followed by BPD 5%, BED 0.5%, BPD 1%, and BED 0.1% added Yukwa. The expansion ratios of BPD 5% and BED 1% added Yukwa remarkably decreased compared to those of non-added and other treatments. The sensory evaluation values corroborated the results of the Yukwa expansion ratio. The acid values of Yukwa under high temperature storage ($60{\pm}1^{\circ}C$) increased rapidly after 20 days in all treatments, and those of BPD 5% and 1% added Yukwa increased slowly compared to non-added Yukwa. ABTS radical scavenging activities of Yukwa showed little change during storage in all treatments. As a results, addition of sorghum bran below BPD 1% and BED 0.5% was suitable for antioxidant activity, quality characteristics and sensory evaluation.

EFFECT OF LASER IRRADIATION AND FLUORIDE APPLICATION ON REMINERALIZATION OF ERODED PRIMARY DENIAL ENAMEL (침식된 유치 법랑질에 대한 레이저 조사 및 불소도포의 재광화 효과)

  • Yang, Young-Sook;Kim, Dae-Eup;Ra, Ji-Young;Lee, Kwang-Hee
    • Journal of the korean academy of Pediatric Dentistry
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    • 제33권2호
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    • pp.262-268
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    • 2006
  • The purpose of this in vitro study was to evaluate the remineralizing effects of Nd : YAG laser irradiation and fluorides application on primary tooth enamel eroded by acidic drink. The materials were 30 sound primary teeth with intact smooth enamel surfaces. They were demineralized with Coca-cola at $37^{\circ}C$ for 12 hours and then irradiated by Nd: YAG laser with 6W power, $50mJ/cm^2$ energy density. and 20Hz pulse repetition. After laser irradiation, teeth were treated by three kinds of fluorides : (1) 0.05% NaF solution, (2) 1.23% APF gel and (3) 0.1% fluoride varnish. Diagnodent scores and microhardness (VHN) were measured before and after the each treatment. The results were as follows: 1. Diagnodent scores decreased to 23.51% from the initial after demineralization, and then increased to 37.37% after laser irradiation, and to 51.34% after fluoride treatment. There were significant differences between the total scores of initial, demineralization, laser irradiation and fluoride treatment (P<0.05). There was no significant difference between scores after fluoride treatment according to fluoride types. 2. Microhardness(VHN) decreased to 33.58% from the initial after demineralization and then increased to 43.99% after laser irradiation, and to 51.38% after fluoride treatment. There were significant differences between the total scores of initial, demineralization, laser irradiation and fluoride treatment (P<0.05). There was no significant difference between scores after fluoride treatment according to fluoride types.

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Establishment of a Murine Model for Radiation-induced Bone Loss in Growing C3H/HeN Mice (성장기 마우스에서 방사선 유도 골소실 동물모델 확립)

  • Jang, Jong-Sik;Moon, Changjong;Kim, Jong-Choon;Bae, Chun-Sik;Kang, Seong-Soo;Jung, Uhee;Jo, Sung-Kee;Kim, Sung-Ho
    • Journal of Radiation Protection and Research
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    • 제40권1호
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    • pp.10-16
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    • 2015
  • Bone changes are common sequela of irradiation in growing animal. The purpose of this study was to establish an experimental model of radiation-induced bone loss in growing mice using micro-computed tomography (${\mu}CT$). The extent of changes following 2 Gy gamma irradiation ($2Gy{\cdot}min^{-1}$) was studied at 4, 8 or 12 weeks after exposure. Mice that received 0.5, 1.0, 2.0 or 4.0 Gy of gamma-rays were examined 8 weeks after irradiation. Tibiae were analyzed using ${\mu}CT$. Serum alkaline phosphatase (ALP) and biomechanical properties were measured and the osteoclast surface was examined. A significant loss of trabecular bone in tibiae was evident 8 weeks after exposure. Measurements performed after irradiation showed a dose-related decrease in trabecular bone volume fraction (BV/TV) and bone mineral density (BMD), respectively. The best-fitting dose-response curves were linear-quadratic. Taking the controls into accounts, the lines of best fit were as follows: BV/TV (%) = $0.9584D^2-6.0168D+20.377$ ($r^2$ = 0.946, D = dose in Gy) and BMD ($mg{\cdot}cm^{-3}$) = $8.8115D^2-56.197D+194.41$ ($r^2$ = 0.999, D = dose in Gy). Body weight did not differ among the groups. No dose-dependent differences were apparent among the groups with regard to mechanical and anatomical properties of tibia, serum ALP and osteoclast activity. The findings provide the basis required for better understanding of the results that will be obtained in any further studies of radiation-induced bone responses.

Clinical Implication of Serum TNF-$\alpha$ and IL-1$\beta$ Measurement in Patients with Sepsis (패혈증환자에서 혈청 TNF-$\alpha$ 및 IL-1$\beta$)

  • Kim, Jae-Yeol;Choi, Hyung-Seok;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Min, Kyung-Up;Kim, Yoo-Young;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • 제49권2호
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    • pp.217-224
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    • 2000
  • Background : It is well known that when macrophages are stimulated with endotoxin, they produce a wide variety of cytokine mediators, including TNF-$\alpha$ and IL-1$\beta$. However, there is an alteration in the macrophages' responsiveness when they are challenged with repeated bouts of endotoxin, termed "endotoxin tolerance" which is regarded as a self-protective phenomenon from continuous stimulation. In this study, endotoxin tolerance in the peripheral blood monocytes of sepsis patients was evaluated. Methods : Fourteen patients with organism-documented sepsis were included. The severity of illness was evaluated by APACHE II score. Peripheral blood monocytes were isolated from the patients and diluted to $1{\times}10^5$ well. After stimulation with endotoxin (LPS of E. coli O114 : B4, 100 ng/ml), they were incubated at $37^{\circ}C$ in 5% $CO_2$ incubator for 24 hours. Supernatant was collected for the measurement of TNF-$\alpha$ and IL-1$\beta$ with ELISA method. Peripheral blood monocytes of seven healthy volunteers were used as control. Results : The APACHE II score (mean$\pm$SD) of the patients at the time of blood sampling was 12.2$\pm$5.7. The primary infection foci were urinary tract infection, pneumonia, subacute bacterial endocarditis, and catheter related infection, etc. The causative organisms were gram negative rods (10 cases), gram positive cocci (6 cases) with two cases of mixed infection. Serum TNF-$\alpha$ could be measured in 4 cases with 29.9$\pm$27.7 pg/ml. Serum IL-1$\beta$was measurable in only one patient. The TNF-$\alpha$ level of supernatant of cultured peripheral blood monocytes was 2,703$\pm$2,066 pg/ml in patients and 2,102$\pm$1914 pg/ml in controls. The IL-1$\beta$level of supernatant was 884$\pm$1,050 pg/ml in patients and 575$\pm$558 pg/ml in controls. There was no difference of TNF-$\alpha$ and IL-1$\beta$ level between patients and controls. Conclusion : We cannot prove the phenomenon of endotoxin tolerance in this study. Future study needs to be focused on the more severe sepsis patients who were taken for sampling earlier. Addition of serum to the culture medium could be an another valuable option for the success of this study.

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Eotaxin mRNA Expression in Bronchial Mucosa of Patients with Asthma (천식 환자의 기관지 조직에서 Eotaxin mRNA 발현에 관한 연구)

  • In, Kwang-Ho;Cho, Jae-Yun;Kang, Sae-Yong;Lee, Sang-Youb;Shim, Jae-Jeong;Kang, Kyung-Ho;Yoo, Se-Hwa;Na, Young-Soon;Kim, Han-Gyum
    • Tuberculosis and Respiratory Diseases
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    • 제45권4호
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    • pp.697-704
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    • 1998
  • Background: Asthma is a chronic inflammatory disease of the airways characterized by a marked infiltration of eosinophils in the bronchial mucosa. Asthmatic bronchial mucosa produces many factors described as being chemotactic for inflammatory cells. IL-5, RANTES, and MCP-1 alpha are the chemotactic factors for eosinophils, but their roles are controversial. Recently eotaxin that is a potent eosinophil chemoattractant cytokine was detected in a guinea-pig model of allergic airway inflammation, and human eotaxin was cloned. Eotaxin is a specific chemoattractant for eosinophils, but its role in asthma is not confirmed. We examined the in vivo expression of eotaxin in bronchi of asthmatic patients. Methods : 11 asthmatics and 2 normal controls were enrolled. All subjects were underwent bronchoscopy with bronchial biopsies in 2nd or 3rd carina. RNA extraction from biopsy samples was done by acid-guanidium method. Semi-quantitaive RT-PCR was done for evaluation of eotaxin mRNA expression The extent of eosinophil infiltration was evaluated by counting the eosinophils in submucosa in HPF of microscope. Results : Eotaxin mRNA expressed in symptomatic, uncontrolled asthma. Steroid inhibited expression of eotaxin mRNA in asthma. Expression of eotaxin mRNA correlated with eosinophil infiltration in bronchial tissues. Conclusion: Expression of eotaxin mRNA increases in uncontrolled asthma and eotaxin is involved in the recruitment of eosinophils.

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