• Title/Summary/Keyword: eosinophil

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The Effects of Prunus Armeniaca Linne Var Fractions on Th2 Cytokine Expression and Atopic Dermatitis of NC/Nga Mouse (행인(杏仁) 분획물이 Th2 cytokine 발현과 NC/Nga mouse의 아토피 피부염에 미치는 영향)

  • Kang, Ki Yeon;Han, Jae Kyung;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.30 no.4
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    • pp.29-59
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    • 2016
  • Objectives PRAL (Prunus armeniaca Linne Var) has been known to suppress allergic reaction. However, the cellular target and its mechanism of action were unclear. This study was designed to investigate the effect of PRAL on RBL-2H3 mast cell, which is PMA-Ionomycin-induced activated in vitro and the effect of PRAL on the MNC/Nga mice that are DNCB-induced activated in vivo. Methods In this study, IL-4, IL-13 production were examined by ELISA analysis; IL-4, IL-13, IL-31, IL-31Ra, $TNF-{\alpha}$ and GM-CSF mRNA expression were examined by Real-time PCR; manifestations of AP-1 and MAPKs transcription factors were examined by western blotting in vitro. Then skin rashes have been evaluated and verified the distribution of mast cells by H&E and toluidine blue. Also, WBC, eosinophil and neutrophil, IgE level in serum, $IFN-{\gamma}$, IL-4, IL-5 in the splenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$, $CD3^+CD69^+$ in the Axillary Lymph Node (ALN), PBMCs and dorsal skin and IL-5, IL-13, IL-31, IL-31Ra in the dorsal skin by Real-time PCR were all evaluated from the NC/BNga mice. Results As a result of this study, the mRNA expression of IL-4, IL-13, IL-31, IL-31Ra and $TNF-{\alpha}$ and IL-4, IL-13 production, shown in ELISA analysis, were suppressed by PRAL. Results from the western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including AP-1 and p-JNK, p-ERK. Histological examination showed that infiltration levels of immune cells in the skin of the AD-induced NC/Nga mice were improved by PRAL orally adminstration. Orally- administered PRAL group also showred decreased level of IgE in the serum. This group has shown decreased the level of IL-4, IL-5, but shown elevated $IFN-{\gamma}$ level in the splenocyte culture supernatant. The same group also has shown decreased cell numbers of $CD4^+$, $CD8^+$, $CD3^+CD69^+$ in the ALN, and $CD4^+$, $Gr-1^+CD11b^+$ in the dorsal skin. PRAL oral adminstration increased cell numbers of $CD4^+$, but decreased cell numbers of $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$ in the PBMCs. Conclusions Obtained results suggest that PRAL can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis (AD) induced in the NC/Nga mice. This may play an important role in recovering AD symptoms and suppressing pruritus.

Hu.4-1BB-Fc fusion protein inhibits allergic inflammation and airway hyperresponsiveness in a murine model of asthma

  • Kim, Byoung-Ju;Kwon, Ji-Won;Seo, Ju-Hee;Choi, Won-Ah;Kim, Young-Jun;Kang, Mi-Jin;Yu, Jin-Ho;Hong, Soo-Jong
    • Clinical and Experimental Pediatrics
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    • v.54 no.9
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    • pp.373-379
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    • 2011
  • Purpose: 4-1BB (CD 137) is a costimulatory molecule expressed on activated T-cells. Repression by 4-1BB is thought to attenuate Th2-mediated allergic reactions. The aim of this study was to investigate the effect of 4-1BB on allergic airway inflammation in a murine asthma model. Methods: BALB/c mice were sensitized to and challenged with ovalbumin (OVA). Hu.4-1BB-Fc was administered 1 day before the first OVA sensitization or 1 day after the second OVA sensitization. Following antigen challenge, airway responsiveness to methacholine was assessed and bronchoalveolar lavage (BAL) fluid was analyzed. Total immunoglobulin (Ig) E, OVA-specific IgE, $IgG_1$, and $IgG_{2a}$ levels in sera were measured by enzyme-linked immunosorbent assay. Lung pathology was also evaluated. Results: In mice treated with Hu.4-1BB-Fc before the first OVA sensitization, there was a marked decrease in airway hyperresponsiveness, total cell count, and eosinophil count in the BAL fluid. In addition, Hu.4-1BB-Fc treatment decreased serum OVA-specific $IgG_1$ levels and increased serum $IgG_{2a}$ level significantly compared with the corresponding levels in mice sensitized to and challenged with OVA. Hu.4-1BB-Fc-treated mice also showed suppressed peribronchial and perivascular inflammatory cell infiltration. In contrast, treatment with Hu.4-1BB-Fc 1 day after sensitization had no effect on airway hyperresponsiveness and showed less suppression of inflammation in lung tissue. Conclusion: Administration of Hu.4-1BB-Fc can attenuate airway inflammation and hyperreactivity in a mouse model of allergic airway inflammation. In addition, administration before sensitization may be more effective. These findings suggest that 4-1BB may be a useful therapeutic molecule against asthma.

Cell Yield of Cerebrospinal Fluid Cell Count Using Cytocentrifuges

  • Koo, Bon-Kyung;Shim, Hyun-Seol;Oh, Jung-A;Lee, Yong-Tag;Choi, Dae-Yong;Lee, Beom-Se;Kim, Eun-Jee;Lee, Seung-Tae;Kim, Sun-Hee
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.1
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    • pp.37-42
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    • 2013
  • The cells are concentrated approximately 20-fold by cytocentrifugation. This study evaluated the nucleated cell number for cells recovered on slide by using Cytospin-3 (Thermo Shandon Ltd. UK) and Cytopro-7620 (Wescor Inc., USA) cytocentrifuges to hematocytometer cell count of $0{\sim}5WBCs/{\mu}L$ of hematocytometer in the cerebrospinal fluid cell count. One hundred forty eight samples of $0{\sim}5WBCs/{\mu}L$ on hematocytometer, were cytocentrifuged by Cytospin-3 and Cytopro-7620 instruments. The nucleated cell number for cells recovered on slide was counted after Wright stain. The nucleated cell number for cells recovered on slide was 0~40 cells in the 44 samples of $0WBC/{\mu}L$, and 3~95 cells in the 31 samples of $1WBC/{\mu}L$. It was observed that the nucleated cell number for cells recovered on slide was 13~100 cells in the 44 samples of $2WBCs/{\mu}L$, and more than 100 cells in the 29 samples of $3{\sim}5WBCs/{\mu}L$, respectively. In addition, extremely normal lymphocyte, monocyte and polymorphonuclear neutrophil were observed in the 143 samples of $0{\sim}5WBCs/{\mu}L$. Macrophage and eosinophil were also rarely observed. The nucleated cell number for cells recovered on slide was 20 cells, which were regarded as $1WBC/{\mu}L$ in body fluid cell count. However, in this study, we made alterations to report nucleated cell percentage as 0% without preparing the cytocentrifuged slide at $0WBC/{\mu}L$ by using the cell yield in a comparison between the value of $0{\sim}5WBCs/{\mu}L$ and nucleated cell number for cells recovered on slide.

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The Effects of Forsythiae Fructus n-BuOH Fraction on Atopic Dermatitis (연교(連翹) n-BuOH 분획물의 아토피 피부염 억제 효과)

  • Lee, Jin Hwa;Han, Jae Kyung;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.30 no.3
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    • pp.1-30
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    • 2016
  • Objectives Previous studies have found out that Forsythiae Fructus (FF) extracts have anti-atopic activities by in vitro experiment. In order to understand more about FF extracts' benefit, we subdivided FF extracts depending on systematic fractionation method by using Methylene chloride (MC), Ethyl acetate (EtOAc), n-BuOH and n-hexane (n-Hx). This study is designed to examine the effect of FF fractions on the PMA- ionomycin-induced activation of RBL-2H3 mast cell lines in vitro and on the DNCB-induced activation of NC/Nga mice in vivo. Methods For this study, we examined IL-4, IL-13 production by ELISA analysis, IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$ mRNA expression by real-time PCR and manifestations of AP-1 and MAPKs transcription factors by western blotting in vitro. Through in vitro experiment, we selected FF n-BuOH fraction that seems the best effective in atopic dermatitis then induced it on NC/Nga mice by DNCB. We measured mice's WBC, eosinophil and neutrophil in heart blood, IL-4, IL-5, IFN-${\gamma}$ in the spleenocyte culture supernatant, the absolute cell numbers of CD4+, CD8+, B220+CD23+, CD3+CD69+ and Gr-1+CD11b+ in the PBMCs, ALN and dorsal skin, IL-5, IL-13, IL-31, IL-31RA in the dorsal skin by real-time PCR and the distribution of immune cells by H&E on dorsal skin and ANL and toluidine blue staining on dorsal skin. Results FF n-BuOH fraction suppressed IL-4, IL-13 production and mRNA expression of IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$. Results from the western blot analysis showed that FF n-BuOH fraction reduced the activation of the mast cell specific transduction factors involved in AP-1 by suppressing JNK and ERK phosphorylation. In the gross, atopic dermatitis induced by DNCB in NC/Nga mice were improved by oral administration of FF n-BuOH fraction. Oral FF n-BuOH fraction also decreased the level of IgE in mice's serum and the level of IL-4 and IL-5 in the spleenocyte culture supernatant, cell numbers of CD8+, B220+CD23+ in the PBMCs, CD4+ in the ALN and CD4+, Gr-1+CD11b+ in the dorsal skin and suppressed mRNA expression of IL-5, IL-13, IL-31, IL-31RA in the dorsal skin. Histological examination showed that infiltration levels of immune cells in atopic dermatitis induced NC/Nga mice were improved by FF n-BuOH fraction. Conclusions FF n-BuOH fraction can reduce pruritus by suppressing IL-31, IL-31RA secretion and modulate molecular mediators and immune cells associated with atopic dermatitis induced in NC/Nga mice which may have played a significant role in recovering atopic dermatitis symptoms.

Relationship between serum 25-hydroxyvitamin D and interleukin-31 levels, and the severity of atopic dermatitis in children

  • Cheon, Bo Ram;Shin, Jeong Eun;Kim, Yun Ji;Shim, Jae Won;Kim, Deok Soo;Jung, Hye Lim;Park, Moon Soo;Shim, Jung Yeon
    • Clinical and Experimental Pediatrics
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    • v.58 no.3
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    • pp.96-101
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    • 2015
  • Purpose: Atopic dermatitis (AD) is a chronic inflammatory relapsing skin disorder. Vitamin D plays a pivotal role in the development of AD, and interleukin (IL) 31 is known to be related to pruritus in AD. The aim of our study was to determine whether 25-hydroxyvitamin D (25(OH)D) levels are related to IL-31 levels or to the severity of AD. Methods: We enrolled 91 children with AD and 32 control subjects without history or symptoms of allergic diseases. Blood was drawn to evaluate complete blood cell count, total eosinophil count (TEC), and total IgE, specific IgE to common allergens, 25(OH)D, and IL-31 levels. Serum 25(OH)D and IL-31 levels were measured using high-performance liquid chromatography and enzyme-linked immunosorbent assay, respectively. The scoring atopic dermatitis (SCORAD) index was used to evaluate the severity of AD. Results: The mean 25(OH)D level was significantly lower in the AD group than in the control group; 25(OH)D decreased greatly in the moderate and severe AD groups compared with the mild AD group. Children with atopic sensitization showed significantly lower 25(OH)D levels than nonatopic children. However, serum IL-31 levels were not related to AD group, SCORAD index, or 25(OH)D levels. The SCORAD index was inversely correlated with serum 25(OH)D level and positively correlated with TECs and total IgE levels. Children with moderate and severe AD had significantly higher TECs than children with mild AD. Conclusion: Vitamin D is related to the severity of AD independently of IL-31.

Novel glutathione-containing dry-yeast extracts inhibit eosinophilia and mucus overproduction in a murine model of asthma

  • Kim, Yun-Ho;Choi1, Yean-Jung;Lee, Eun-Jung;Kang, Min-Kyung;Park, Sin-Hye;Kim, Dong Yeon;Oh, Hyeongjoo;Park, Sang-Jae;Kang, Young-Hee
    • Nutrition Research and Practice
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    • v.11 no.6
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    • pp.461-469
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    • 2017
  • BACKGROUND/OBSECTIVE: Airway inflammation by eosinophils, neutrophils and alveolar macrophages is a characteristic feature of asthma that leads to pathological subepithelial thickening and remodeling. Our previous study showed that oxidative stress in airways resulted in eosinophilia and epithelial apoptosis. The current study investigated whether glutathione-containing dry yeast extract (dry-YE) ameliorated eosinophilia, goblet cell hyperplasia and mucus overproduction. MATERIALS/METHOD: This study employed $2{\mu}g$/mL lipopolysaccharide (LPS)- or 20 ng/mL eotaxin-1-exposed human bronchial epithelial cells and ovalbumin (OVA)-challenged mice. Dry-YE employed in this study contained a significant amount of glutathione (140 mg in 100 g dry yeast). RESULTS: Human bronchial epithelial cell eotaxin-1 and mucin 5AC (MUC5AC) were markedly induced by the endotoxin LPS, which was dose-dependently attenuated by nontoxic dry-YE at 10-50 ${\mu}g$/mL. Moreover, dry-YE inhibited the MUC5AC induction enhanced by eotaxin-1, indicating that eotaxin-1-mediated eosinophilia may prompt the MUC5AC induction. Oral supplementation with 10-100 mg/kg dry-YE inhibited inflammatory cell accumulation in airway subepithelial regions with a reduction of lung tissue level of intracellular adhesion molecule-1. In addition, ${\geq}50$ mg/kg dry-YE diminished the lung tissue levels of eotaxin-1, eosinophil major basic protein and MUC5AC in OVA-exposed mice. Alcian blue/periodic acid schiff staining revealed that the dry-YE supplementation inhibited goblet cell hyperplasia and mucus overproduction in the trachea and bronchiolar airways of OVA-challenged mice. CONCLUSIONS: Oxidative stress may be involved in the induction of eotaxin-1 and MUC5AC by endotoxin episode and OVA challenge. Dry-YE effectively ameliorated oxidative stress-responsive epithelial eosinophilia and mucus-secreting goblet cell hyperplasia in cellular and murine models of asthma.

Inhibitory effects of Paridis Rhizoma in the activation and proliferation of eosinophils: implications on its regulatory roles for asthma (rmIL-5로 유도된 호산구의 활성화 및 성장에서 중루의 천식반응 억제효과)

  • Sin, Mi-Kyung;Kil, Ki-Jeong;Lee, Young-Chul;Kim, Jin-Suk;Seo, Young-Bae;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.20 no.2
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    • pp.159-169
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    • 2005
  • Objectives : This study was carried out for the purpose of knowing the effect from anti-arthma action of the abstraction from a extract of Paridis Rhizoma(EPR). In order to know what the effect of controlling an abstraction from Paridis Rhizoma. and about the expression of B cells and Ig E cells, mast cells it was necessary for it to be activated by ovalbumin. Methods : In order to know what the effect was on the organization of cytokine gene expression from The increase and divorce of the B cells and allergic acting by EPR, we found it necessary to examine the BALF. At the same time, as we examined the histamine release by ELISA method, we also examined the effect of EPR. Results : EPR at $100\;{\mu}g/ml$, the highest concentration examined did not have any cytotoxic effects on mLFCs. In FACS analysis, number of granulocyte/lymphocyte, $CD3e^+/CCR3^+,\;CD4^+\;and\;CD23^+/B220^+$ in asthma-induced lung cells were significantly decreased by EPR treatment compared to the control group. In RT-PCR analysis, mRNA expression for CCR3, eotaxin and histamine in asthma-induced lung cells, which was induced by rIL-3 plus rmIL-5 treatments, was significantly decreased by EPR treatment. In ELISA analysis, production levels of IL-4, IL-13 and histamine in asthma-induced lung cells, which were induced by rIL-3 plus rmIL-5 co-treatment, were significantly decreased by EPR treatment. EPR treatments significantly inhibited the proliferation of eosinohils prepared from asthma-induced mouse lung tissues compared to the non-EPR treated control cells. Immunohistochemical analysis revealed that EPR treatment significantly decreased the levels of eosipnphil activation compared to non-treated cells. Conclusion : The present data suggested that Paridis Rhizoma may have an effects on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of Paridis Rhizoma.

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Interleukin-5 Inhibition Assay of the Oriental Materia Medica Treatment by High Through Screening on the Splenocyte of Mouse (High through screening(HTS)를 이용한 백서(白鼠)의 비장세포(脾臟細胞)에서의 IL-5 분비 조절 한약물(韓藥物) Screening)

  • Park, Dong-Hee;Rhee, Hyung-Koo;Jung, Sung-Ki;Jung, Hee-Jae
    • The Journal of Internal Korean Medicine
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    • v.27 no.2
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    • pp.394-406
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    • 2006
  • Backgrounds and Objectives: Asthma is considered to be an inflammatory disease characterized by airway hyperresponsiveness and Pulmonary eosinophilia. And it is known the structure and function of IL-5, its receptor and the mechnism IL-5 triggered eosinophil accumulation and inflammaion of the airways. At this point of view, we assume which oriental materia medics can the splenocyte inhibit from secreting the IL-S in vitro. Material and Methosds: We used the splenocyte of mouse 8 weeks after its birth, and then cnltivated those into the 2 experimental groups and control group for 48 hours. The culture medium of experimental groups were made of $1{\mu}g/ml,\;10{\mu}g/ml$, oriental materia medics, representative. And the culture media of control group was given no oriental materia medica. Then, we assayed the quantity of cytokine-expression by the Sandwich ELISA. The quantifies of cytokine-expression of the experimental groups were compared with that of the control group which was standardized These method were used for the all of oriental materia medica treated. Results: In this study, we demonstrated that 12 oriental materia medica that inhibit the splenocyte from secreting the IL-5 in both $1{\mu}g/ml,\;and\;10{\mu}g/ml$ culture media. Those were Equiseti Herbs, Sophorae Subprostratae Radix, Moutan Radicis Cortex. Trichosanthis Radix, Buddleiae Flos. Cyperi Rhizoma. Benincasae Semen, Armeniacae Semen. Zedoariae Rhizoma, Astragali Semen, Dolichoris Semen. Lilii Bulbus, Asparagi Radix, Atractylodes Rhizoma White, Polygonati Officinallis Rhizoma. Conculusions: These findinga indicate that some oriental materia medica, specially Antipyretics, Herbs for Resolving Phlegm, Relieving Cough and Calming Wheezing and Herbs for Tonifring and Invigorating effects inhibit the splenocyte from secreting the IL-5. And further study experimented in vivo is needed for treating IL-5-driven inflammatory disease including asthma.

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Studies on the Blood Picture of Holstein Cows in Korea (Holstein 암소의 혈액상(血液像)에 관하여)

  • Mun, Hi Cheol;Choi, Hi In;Cheong, Chang Kook
    • Korean Journal of Veterinary Research
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    • v.14 no.1
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    • pp.9-16
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    • 1974
  • The literature on the hematology of dairy cattle contains very important information of physiological and clinical field, but a comprehensive survey on the blood picture of Holstein cows has not been made yet in Korea. The object of the present investigation was to make good this deficiency, and to suggest standards for the blood picture of Holstein cows kept under average farming condition in this country. Observations were made on the blood picture of 30 healthy non-pregnant Holstein cows aging from 6 to 10 years. All of them were selected at random from 15 livestock farms of Deajeon area in order to determine the hematological values from January to Feburary, 1974. The ranges and mean values of erythrocyte count, hemoglobin in blood, hematocrit value, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and total and differential leukocyte count were determined. The result obtained in this work were summerized as follows: 1. Ranges and mean values(with standard error) of erythrocyte count, hemoglobin in blood and hematocrit values were 4.90~7.82 and $5.83{\pm}0.12{\times}10^6/mm^3$, 7.8~10.3 and $8.7{\pm}0.11g/100ml$, and 26~38 and $30.2{\pm}0.53$ mI/100 ml, respectively. 2. Mean corpuscular hemoglobin, mean corpuscular volume, and mean corpuscular hemoglobin concentration showed ranges and mean values(with standard error) of 13.2~17.4 and $14.8{\pm}0.22$ pg, 46.0~71.8 and $52.3{\pm}0.31{\mu}m^3$, and 21.6~32.7 and $28.6{\pm}0.47$ g/100 ml, respectively. 3. Total leukocyte count showed a range of 6,300~13,600 and a mean value(with standard error) of $10,035{\pm}346/mm^3$. 4. Ranges and mean values(with standard error) of differential leukocyte counts of total neutrophil were 21~40 and $36.4{\pm}0.7%$, 1,638~4,080 and $3,233{\pm}111/mm^3$, of band neutrophil 0~9 and $3.9{\pm}0.5%$, 0~1,028 and $390{\pm}54/mm^3$, of segmented neutrophil 18~35 and $28.4{\pm}0.7%$, 1,386~3,672 and $2,842{\pm}103/mm^3$, of lymphocyte 46~69 and $55.3{\pm}1.1%$, 3,380~8,976 and $5,535{\pm}263/mm^3$, of monocyte 0~4 and $1.7{\pm}0.2%$, 0~324 and $116{\pm}21/mm^3$, of eosinophil 4~15 and $10.8{\pm}0.6%$, 540~1,725 and $1,082{\pm}72/mm^3$. No basophil was observed in this work.

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Research on Outcome Indicators in Clinical Trials of Asthma, Cough, and Sputum by Reviewing Papers Searched from PubMed (해외논문고찰을 통한 천식 및 기침${\cdot}$가래 치료제의 유효성 평가지표 개발연구)

  • Oh, Mi-Hyune;Lee, Ji-Hyung;Jang, Bo-Hyoung;Jung, Sung-Ki;Sun, Seung-Ho;Ko, Ho-Yeon;Jun, Chan-Yong;Park, Jong-Hyeong;Ko, Seong-Gyu
    • The Journal of Internal Korean Medicine
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    • v.28 no.3
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    • pp.519-530
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    • 2007
  • Objectives : The purpose of this study was to develop outcome indicators in clinical trials of herbal drugs effective for asthma. cough and sputum. To accomplish the objective, this study collected outcome indicators developed and used according to conventional medical concepts. Methods : Our research group reviewed SCI papers concerned with developing outcome indicators to evaluate amelioration of asthma, cough and sputum. We also reviewed clinical trials of herbal drugs effective for them. Results : To evaluate asthma, objective as well as subjective methods were chosen according to the purpose of each trial. Objective methods were PEF, FEVl, serum IgE, peripheral eosinophil counts, and so on. Subjective methods were symptom scores, symptom diaries, quality of life measures, etc. To evaluate cough and sputum, objective and subjective methods were also chosen. Objective methods were tussigenic challenges, sputum induction and computerized methodology, and subjective methods were similar to the methodology evaluating asthmatic symptoms. Conclusions : It is desirable for a clinical trial evaluating herbal drugs for asthma, cough and sputum to use objective and subjective outcome indicators together. However, biological outcome indicators, a kind of objective methods, can not be chosen as the purpose of trial. Valid and reliable subjective outcome indicators are needed to develop good clinical trials of herbal drugs effective for asthma, cough and sputum.

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