• 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.

• 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.

• 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.

• 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.

• 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.

• 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.

• 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.

• 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.

• Journal of Animal Science and Technology
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• v.51 no.4
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• pp.321-328
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• 2009

A total of fourteen, 1-wk-old male Holstein calves were allotted into two groups consisted of control (CON) and IGY which was orally administrated with immunoglobulin yolk (IgY) for 1wk. Calves in both groups were provided with milk replacer according to feeding program and had ad libitum access to timothy hay for the entire experimental period (7wks). At 0, 7 and 49 day of experiment, blood samples were collected from the jugular vein of calves to investigate blood biochemical profiles and the differential count (%) of white blood cell (WBC). We also monitored growth performance and colony forming unit (CFU) of fecal microbial population in calves. The adminstration of IgY in calves did not affect body weight and weight gain during 49 days feeding trial compared with control group. The CFU of E. coli and Lactobacilli in the feces of calves were not significantly affected by IgY treatment, whereas the score of the calf scours during day 43 to 49 in IgY group showed a significant (P<0.05) solid type. There were no differences in plasma biochemical components including total protein, albumin, immunoglobulin and the other indicators. As for WBC differential count (%), there was no statistical difference in the percentages of neutrophil, lymphocyte, monocyte, eosinophil and basophil at 0, 7 and 49 days after the oral supplementation of IgY. In conclusion, the oral supplementation of IgY as an immunostimulant did not affect growth performance, fecal microbial population, blood biochemical profile and WBC differential count in Holstein calves.

• Korean Journal of Veterinary Research
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• v.30 no.2
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• pp.157-162
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• 1990

The hematologic values were examined from 74 healthy mixed breed dogs in the area of Seoul. The results obtained are summarized as follows; 1. Mean${\pm}$SD values and ranges of red blood cell(RBC) count were $6.16{\pm}0.92{\times}10^6/{\mu}l$ and $4.40{\sim}8.62{\times}10^6/{\mu}l$, of hemoglobin(Hb) content $14.90{\pm}2.42g/100ml$ and 8.7~19.2g/100ml, of packed cell volume(PCV) $45.47{\pm}6.16ml/100ml$ and 30~57ml/100ml, of mean corpuscular volume (MCV) $74.80{\pm}6.83fl$ and 54.08~90.90fl, of mean corpuscular hemoglobin (MCH) $24.41{\pm}3.91pg$ and 14.19~32.97pg, of mean corpuscular hemoglobin concentration (MCHC) $32.07{\pm}3.24g/100ml$ and 22.23~39.76g/100ml, respectively. 2. The RBC count value in the age group of 3~4 years was higher (p<0.05) than the total RBC count value. The Hb content value in the age group of less than 6 months was lower (p<0.01) but in the age group of 1~2 years and the age group of 3~4 years were higher (p<0.05, p<0.01) than the total Hb content value. The PCV value in the age group of less than 6 months was lower (p<0.05) than the total PCV value. The MCHC values in the age group of 1~2 years and the age group of 3~4 years were higher (p<0.01, p<0.05) than the total MCHC value. 3. Mean${\pm}$SD values and ranges of white blood cell (WBC) count were $11.26{\pm}3.05{\times}10^3/{\mu}l$ and $6.30{\sim}18.4{\times}10^3/{\mu}l$, of band neutrophil $2.97{\pm}1.44%$ and 1~10%, of segmented neutrophil $62.81{\pm}4.92%$ and 42~70, of lymphocyte $30.55{\pm}5.69%$ and 17~50%, of monocyte $2.49{\pm}0.84%$ and 1~5%, of eosinophil $1.81{\pm}1.175$ and 1~8%, respectively. 4. The WBC count value in the age group of 7~12 months was lower (p<0.05) but in the age group of 3~4 years was higher (p<0.05) than the total WBC count value. The band neutrophil values in the age group of 1~2 years and the age group of 3~4 years were higher (p<0.05, p<0.01) than the total band neutrophil value.