• Title/Summary/Keyword: Cell analysis

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Post-operative Radiation Therapy for Esophageal Cancer; Analysis of Failure Pattern (식도암의 수술 후 방사선 치료: 실패 양상 분석)

  • Kim Mi Sook;Kim Jae Young;Yoo Seoung Yul;Zo Chul Goo;Yoo Hyung Jun;Zo Jae Ill;Baek Hee Jong;Park Jong Ho;Choi Soo Yong
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.447-454
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    • 1998
  • Purpose : This study evaluated the survival, local control, prognostic factor, and failure pattern of patients with esophageal cancer treated with operation and adjuvant radiation therapy to use as fundermental data of postoperative radiation therapy. Materials and Methods : A retrospective analysis was undertaken of 82 patients who had locally advanced esophageal cancer treated with operation and adjuvant radiation therapy from January 1988 to December 1995. According to AJCC staging, stage IIA were in 26 patients, stage IIB in 4 patients, and stage III in 52 patients. Squamous cell carcinoma were in 77 patients, adenosquamous carcinoma in 3 patients, and adenocarcinoma in 2 patients. The patients received radiation therapy ranging from 41.0 Gy to 64.8 Gy. Five patients received neoadjuvant chemotherapy. Results : Two-year survival and local control rates for all patients were 36.8$\%$ and 30.4$\%$ respectively. And they were 9.3$\%$ and 26.3$\%$ respectively at 5 years. According to stages, 2-year survival rates were 50.2$\%$ in IIA, 0$\%$ in IIB and 23.3$\%$ in III (p=0.004). Two-year local control rates were 49.2 $\%$ in IIA, 66.6$\%$ in IIB and 24.7$\%$ in III (p=0.01). Sixty patients developed recurrence, which were 3 tumor margin, 23 lymph node recurrence, 4 tumor margin and lymph node, 1 tumor margin and distant metastasis, 9 lymph node and distant metastasis, 17 distant metastasis and 3 unknown metastatic site. Prognostic factors affecting survival were smoking (p=0.02), T-staging (p=0.0092), N-staging (p=0.0045). Prognostic factors affecting local control were T-staging (p=0.019), N-staging (p=0.047). Conclusion : In spite of post-operative radiation therapy, predominant failure pattern was local failure. Especially regional lymph node failure was major cause of local failure. So strategy of aggresive adjuvant radiation therapy to regional lymph node area in post operative treatment should be proposed.

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Effects of Chronic and Acute Stress on Clusterin Secretion of the Rat Submandibular Gland (급만성 스트레스가 백서 악하선의 Clusterin 분비에 미치는 영향)

  • Jin, Sang-Bae;Chun, Yang-Hyun;Hong, Jung-Pyo
    • Journal of Oral Medicine and Pain
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    • v.31 no.1
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    • pp.79-89
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    • 2006
  • The aim of this study is to know how the rat submandibular gland changes under various emotional stress condition, using molecular biological methods. Restraint and chronic unpredictable mild stress (CUMS) experiment is conducted on fifty one 7-week old Sprague-Dawley rats (restraint stress experiment: 21, CUMS: 30). The rats were sacrificed, the submandibular glands were excised immediately at certain time, and examined by the use of immunohistochemistry and western blotting. In CUMS experiment, sucrose preference test, water intake change, weight change were implemented at 1 week interval for the experimental period The results are as follows: 1. The number of clusterin-secreting cells of restraint stress group compared to control group showed significantly decreasing tendency in all experimental groups except for the 1st hour group (p<0.001 in the 9th, 24th, 72nd, 120th, and 168th hour group). 2. The number of clusterin-secreting cells of CUMS group compared to control group showed significantly increasing tendency in the 2nd week group (p<0.01), and significantly decreasing tendency in the 4th and 5th week group (p<0.001). 3. Sucrose preference test in CUMS experiment showed significant difference between the 5th week experimental group and control group (p<0.01). 4. Weight change in CUMS experiment showed significant difference between the 5th week experimental group and control group (p<0.01), but water intake change didn't show significant difference compared to control group. 5. In western blot analysis, clusterin expression was decreased on a gradual basis in due time compared to the control group in the restraint stress group. As for CUMS group (chronic unpredictable mild stress group), it was increased till the 2nd week and decreased till the 5th week after that, which is similar to immunohistochemical analysis result and the decreasing tendency of sucrose preference and weigh changes. Through the test, it was proved that expression of clusterin in saliva glands decreases after receiving either acute or chronic stress, indicating relation with depression caused by chronic stress. Unlike other data, however, apoptotic tendency was hardly found in tissues. Diverse possibilities could be suggested on that: first, the stress was not enough to expedite apoptosis; second, apoptosis-related protein was already being secreted though not detected with microscope; third, clusterin, a major secretion molecule of saliva, decreased with saliva's malfunction due to stress. In the respect, it will be necessary to examine proteins expressed in case of cell death or other heat-shock proteins at the same time, in order to see whether any cellular change or death is caused by decreasing clusterin under high stress, and whether the original state is restored as time goes by under mild stress, through longer-term tests using even higher acute stress.

Evaluation of Commercial Korean Honey Quality and Correlation Analysis of the Quality Parameters (국내 시판 벌꿀의 품질 평가 및 품질인자간 상관관계 분석)

  • Sung, Hwa-Jung;Jung, Chuleui;Kwon, Jiyoung;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1489-1500
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    • 2018
  • Honey is made from flower nectar by honey bees. In this study, 120 honeys from various flowers and across eight different provinces in Korea were collected and their components, antioxidants, and hemolytic activities against red blood cell were evaluated. Our results show that total polyphenol (TP) varied widely across the samples, with chestnut honey showing the highest TP ($77.1{\pm}8.4mg/100g$), protein content ($25.9{\pm}0.9mg/100g$), and absorbance at 400 nm ($A_{400}$ : $0.156{\pm}0.036$). In contrast, the acacia honey and sugar honey had a TP of 9.5~30 mg, 12~15 mg/100g of, and the lowest $A_{400}$ of $0.06{\pm}0.02$. High amounts of total flavonoid were quantified in the jujube and chestnut honeys at $8.73{\pm}7.31$ and $8.39{\pm}3.02mg/100g$, respectively. No samples demonstrated hemolytic activity up to 1 mg/ml. Antioxidant activities determined by DPPH, ABTS, and nitrite scavenging placed the chestnut honey highest, followed by jujube, styrax, multi-floral, citrus, acacia and sugar honey. Analysis of parameter correlations indicated that the components and bioactivity of the honey are dependent on the origin of the flower rather than on bee-farming regions. A positive correlation between TP content and antioxidant activity was identified. The correlation coefficients between $A_{400}$ and the TP, ABTS scavenging, and reducing power values were 0.804, 0.772 and 0.741, respectively. We therefore suggest that $A_{400}$ could be used as a noble, economic and simple factor for honey quality evaluation. Our results can potentially be used to develop functional honey for the food and pharmaceutical industries.

Antibacterial and Antibiofilm Activities of Leaf Extracts of Stewartia koreana against Porphyromonas gingivalis (Porphyromonas gingivalis에 대한 노각나무 잎 추출물의 항균활성 및 생물막 형성 억제 효과)

  • Kim, Hye Soo;Park, Min Jeong;Kim, Soo Jeong;Kim, Bu Kyung;Park, JunHo;Kim, DaeHyun;Cho, Soo Jeong
    • Journal of Life Science
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    • v.31 no.3
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    • pp.330-337
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    • 2021
  • This study was conducted to investigate the potential of Stewartia koreana as oral healthcare materials. The antibacterial activity of ethanol extracts from leaves and branches of S. koreana against oral bacteria was confirmed. The leaf and branch extracts (1 mg/disc) showed antibacterial activity against P. gingivalis only among several tested oral bacteria. The leaf extracts showed higher antibacterial activity, with values similar to those of chlorhexidine, which was used as a positive control. The MIC of the leaf extract against P. gingivalis was 0.4 mg/ml and showed bacteriostatic action. The inhibitory effects of the extract on biofilm formation and on gene expression related to biofilm formation by P. gingivalis were determined by biofilm biomass staining, scanning electron microscopy (SEM), and qRT-PCR analysis. The biofilm production rate and cell growth of P. gingivalis in the cultures treated with 0.2-2.0 mg/ml of S. koreana leaf extracts were significantly decreased in a concentration-dependent manner. The inhibitory effect on the formation of P. gingivalis biofilms at concentrations of 1 mg/ml was confirmed by SEM. The qRT-PCR analysis showed concentration-dependent suppression of the fimA and fimB gene expression associated with fimbriae formation in the cultures treated with 0.2-2.0 mg/ml S. koreana leaf extract. These results support the conclusion that S. koreana leaf extracts can be used as oral healthcare materials derived from natural materials, as demonstrated by the antibacterial action and inhibition of biofilm formation of P. gingivalis.

Production of doubled haploid population derived from the microspore culture of rapeseed (Brassica napus L.) F1 generation and analysis of fatty acid composition (유채 잡종 1세대의 소포자 배양에 의한 배가반수체 집단 선발 및 지방산 조성 분석)

  • Lee, Ji Eun;Park, Ju Hyun;Kim, Kwang Soo;An, Da Hee;Cha, Young Lok
    • Journal of Plant Biotechnology
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    • v.49 no.1
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    • pp.74-81
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    • 2022
  • Brassica napus, an oil crop that produces rapeseed oil, is an allotetraploid (AACC, 2n = 38) produced by natural hybridization between B. rapa and B. oleracea. In this study, microspore was cultured using the F1 developed from a cross between 'EMS26' line with high oleic acid content and 'J8634-B-30' lines. The flower bud size showing the nuclear development at the late uninucleate and binucleate stage with high embryogenesis rate was 2.6 ~ 3.5 mm. Microspores were cultured using only this size and after then most microspore embryo developed into secondary embryos and then regeneration plants obtained from the developed multilobe. The analysis of the ploidy of the plants revealed that 66.7% and 27.8% of the total lines were tetraploids and octoploids, respectively. The sizes of stomatal cells in tetraploids, octoploids, and diploids were 25.5, 35.6, and 19.9 ㎛, respectively, indicating that ploidy level was positively correlated with cell size. Furthermore, 62 tetraploid doubled haploid (DH) lines were selected. The average oleic acid (C18:1) and linolenic acid (C18:3) concentrations of DH were 72.3% and 6.2%, respectively. Oleic acid and linolenic acid concentrations exceeded the two parental values in 5 and 14 DH lines, respectively, suggesting that these two fatty acids had transgressive segregation. Therefore, the DH population can be utilized for the biosynthesis of unsaturated fatty acids in rapeseed and related genes. It can also be used as a breeding material for varieties with high oleic acid concentrations.

Ginsenoside compound K protects against cerebral ischemia/ reperfusion injury via Mul1/Mfn2-mediated mitochondrial dynamics and bioenergy

  • Qingxia Huang;Jing Li;Jinjin Chen;Zepeng Zhang;Peng Xu;Hongyu Qi;Zhaoqiang Chen;Jiaqi Liu;Jing Lu;Mengqi Shi;Yibin Zhang;Ying Ma;Daqing Zhao;Xiangyan Li
    • Journal of Ginseng Research
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    • v.47 no.3
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    • pp.408-419
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    • 2023
  • Background: Ginsenoside compound K (CK), the main active metabolite in Panax ginseng, has shown good safety and bioavailability in clinical trials and exerts neuroprotective effects in cerebral ischemic stroke. However, its potential role in the prevention of cerebral ischemia/reperfusion (I/R) injury remains unclear. Our study aimed to investigate the molecular mechanism of ginsenoside CK against cerebral I/R injury. Methods: We used a combination of in vitro and in vivo models, including oxygen and glucose deprivation/reperfusion induced PC12 cell model and middle cerebral artery occlusion/reperfusion induced rat model, to mimic I/R injury. Intracellular oxygen consumption and extracellular acidification rate were analyzed by Seahorse multifunctional energy metabolism system; ATP production was detected by luciferase method. The number and size of mitochondria were analyzed by transmission electron microscopy and MitoTracker probe combined with confocal laser microscopy. The potential mechanisms of ginsenoside CK on mitochondrial dynamics and bioenergy were evaluated by RNA interference, pharmacological antagonism combined with co-immunoprecipitation analysis and phenotypic analysis. Results: Ginsenoside CK pretreatment could attenuate mitochondrial translocation of DRP1, mitophagy, mitochondrial apoptosis, and neuronal bioenergy imbalance against cerebral I/R injury in both in vitro and in vivo models. Our data also confirmed that ginsenoside CK administration could reduce the binding affinity of Mul1 and Mfn2 to inhibit the ubiquitination and degradation of Mfn2, thereby elevating the protein level of Mfn2 in cerebral I/R injury. Conclusion: These data provide evidence that ginsenoside CK may be a promising therapeutic agent against cerebral I/R injury via Mul1/Mfn2 mediated mitochondrial dynamics and bioenergy.

Prediction of Necrotizing Pancreatitis on Early CT Based on the Revised Atlanta Classification (개정된 아틀란타 분류법에 근거한 초기 CT에서의 괴사성 췌장염의 예측)

  • Yeon Seon Song;Hee Sun Park;Mi Hye Yu;Young Jun Kim;Sung Il Jung
    • Journal of the Korean Society of Radiology
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    • v.81 no.6
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    • pp.1436-1447
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    • 2020
  • Purpose To investigate the clinical and CT features at admission to predict the progression to necrotizing pancreatitis (NP) in patients initially diagnosed with interstitial edematous pancreatitis (IEP). Materials and Methods Patients with IEP who underwent contrast-enhanced CT at admission and follow-up CT (< 14 days) were included (n = 178). Two radiologists performed a consensus review of follow-up CT scans and diagnosed the type of acute pancreatitis as IEP or NP. Laboratory findings at admission were recorded. Clinical, CT, and laboratory findings were compared between the IEP-IEP group and IEP-NP group using the chi-square test and the t-test. Multivariate analysis was also performed. Results There were 112 and 66 patients in the IEP-IEP and the IEP-NP groups, respectively. The proportion of patients with alcohol etiology was significantly larger in the IEP-NP group. Among the CT findings, the presence of peripancreatic fluid and heterogeneous parenchymal enhancement were more frequently observed in the IEP-NP group. Among the laboratory variables, serum C-reactive protein levels and white blood cell counts were significantly higher in the IEP-NP group. Multivariate analysis revealed that the presence of peripancreatic fluid and heterogeneous parenchymal enhancement were significant findings distinguishing the two groups. Conclusion CT findings, such as the presence of peripancreatic fluid and heterogeneous pancreatic parenchymal enhancement, may be helpful in predicting the progression to NP in patients initially diagnosed with IEP.

The Clinical Significance of ${\gamma}{\delta}$ T lymphocytes in patients with pleural tuberculosis (결핵환자에서 말초혈액과 흉막액내 ${\gamma}{\delta}$ T 림프구의 의의)

  • Song, Kwang Seon;Shin, Kye Chul;Kim, Do Hun;Hong, Ae Ra;Kim, Hee Seon;Yong, Suk Joong
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.1
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    • pp.44-51
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    • 1997
  • Background : The changes of the composition in the T-lymphocyte are important as an immunological abnormality in the pathogenesis of tuberculosis. Previously, the second type of TCR dimer(${\gamma}{\delta}$ T lymphocyte) that did not express CD4 or CD8 molecules was found. In other reports the presence of this type of lymphocytes was increased in the initial stage of tuberculous infections. Method : To determine whether there are some differences in the T-lymphocyte subsets in the peripheral blood or pleural effusion between pleural tuberculosis and other pleurisy. Thirty patients with pleural effusion among the forty-nine patients were examined T-lymphocyte subset analysis(CD4+T-cell,CD8+ T-cell,${\gamma}{\delta}$ T-lymphocytes) with anti- Leu4, anti-Leu3a, anti-Lea2a, anti HLA-DR and anti-TCR-${\gamma}{\delta}$-1(Becton & Dickinson Co.). Results : The average age of the patients was 50 years old(17-81year). There were 33 males and 16 female patients. Patiensts with tuberculosis are 30cases(tuberculous pleurisy 15), lung cancer 12cases(malignant effusion 9) and pneumonia 7cases(parapneumonic effusion 6cases) In T lymphocyte subsets of pleural effusion, helper T lymphocyte(54.6 + 13.8 %) of tuberculous pleurisy was higher than that(36.2 + 25.3 %) of non-tuberculous pleurisy(p=0.04). The peripheral blood ${\gamma}{\delta}$ T-lymphocytes in tuberculousis was insignificantly higher than non-tuberculous patients(p= 0.24). The peripheral blood ${\gamma}{\delta}$ T-lymphocytes and pleural ${\gamma}{\delta}$ T-Iymphocytes in tuberculous pleurisy was insignificantly higher than in non-tuberculous pleurisy(p= 0.16, p= 0.12). Conclusion : The percentage of -${\gamma}{\delta}$ T lymphocytes among the total T-lymphocytes is not significantly increased in the peripheral blood or pleural effusion of the pleural tuberculosis. ${\gamma}{\delta}$ T lymphocytes is less useful as a diagnostic method of pleural tuberculosis.

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Measuring Intracellular Mycobacterial Killing Using a Human Whole Blood Assay (인체 전혈 모델을 이용한 세포내 결핵균 살균력에 관한 연구)

  • Cheon, Seon-Hee;Song, Ho-Yeon;Lee, Eun-Hee;Oh, Hee-Jung;Kang, In-Sook;Cho, Ji-Yoon;Hong, Young-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.5
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    • pp.497-509
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    • 2002
  • Background : The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. Methods : The PPD positive subject were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis $H_{37}Ra$ for 4 days by rotating the culture in a $37^{\circ}C$, 5% $CO_2$ incubator. In some experiments, methlprednisolone- or pentoxifyline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The ${\Delta}$ log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. Results : 1. A trend was noted toward the improved killing of mycobacteria in PPD+ subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifyline adversely affected the killing in the PPD+ subjects umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis $H_{37}Ra$ by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis $H_{37}Ra$. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the ${\Delta}$ logKR continuously decreased in a 3 and 4 days of whole blood culture. Conclusion : The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.

Plasma G-CSF and GM-CSF Concentrations and Expression of their Receptors on the Granulocyte in Children with Leukocytosis (백혈구 증가증 환아의 혈장내 G-CSF와 GM-CSF의 농도 및 과립구에서의 이들 수용체의 발현)

  • Choi, Won Seok;Ryu, Kyung Hwan;Kim, You Jeong;Kim, So Young;Kim, Hyun Hee;Lee, Wonbae
    • Clinical and Experimental Pediatrics
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    • v.46 no.3
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    • pp.271-276
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    • 2003
  • Purpose : Granulocyte-colony stimulating factor(G-CSF) and granulocyte macrophage-colony stimulating factor(GM-CSF) are principal cytokines in granulopoiesis and their physiologic effects are mediated through binding to specific cell surface receptors. Although it is known that the level of serum G-CSF and GM-CSF, and presentation of the receptors are increased in infectious diseases, there have been no studies to find the correlation between the granulopoiesis and leukocytosis. This study was designed to measure G-CSF and GM-CSF in leukocytosis and in control and to demonstrate the possible pathogenesis of granulopoiesis in leukocytosis using quantitative analysis of G-CSF, GM-CSF and their CSFr. Methods : The plasma levels of G-CSF, GM-CSF of 13 children without leukocytosis and 14 children with leukocytosis were measured. Counts of cell surface G-CSFr and GM-CSFr were measured by combining anti G-CSFr and anti GM-CSFr monoclonal antibodies to their respective receptors by using quantitative flow cytometric assay. Results : There was no significant difference betweeen the plasma concentration of G-CSF and GM-CSF in acute leukocytosis and in the control group. However, levels of G-CSFr in acute leukocytosis decreased significantly compared to the control(P=0.012) and the levels of GM-CSFr in both groups revealed no significant difference. Conclusion : Increase in the number of leukocyte in leukocytosis was mediated by increasing the number of neutrophil, and increased plasma concentration of G-CSF may be the cause of neutrophilia. But GM-CSF did not have any influence on leukocytosis.