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Antitumor and Immuno-potentiating Activities of Crude Polysaccharides from Fruiting Body of Agaricus brasiliensis (신령버섯(Agaricus brasiliensis) 자실체 추출 조다당류의 항암 및 면역증강 작용)

  • Cha, Youn-Jeong;Kim, Jeong-Hwa;Lee, Tae-Soo;Lee, U-Youn
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.57-67
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    • 2011
  • Agaricus brasiliensis, one of edible mushroom belonging to Basidiomycota, has been used for curing gastric ulcer and stomach cancer of human beings and also known to have good inhibitory effects on sarcoma 180 and Ehrlich carcinoma of mice. Neutral saline soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were prepared from fruiting body of the mushroom. ${\beta}$-glucan and total protein contents were identify from fractions of edible mushrooms extract. The ${\beta}$-glucan and protein contents of all fractions of the mushrooms ranged from 21.54~32.31% and 0.16~9.34%, respectively. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180, HT-29, NIH3T3 and RAW 264.7 at the concentration of 10~2000 ${\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of 18.8~50.6% in mice previously inoculated with Sarcoma 180. Fr. HW increased the numbers of spleen cell by 1.2 fold at the concentration of 200 ${\mu}g/ml$ compared with control. Fr. MeOH and Na improved the immuno-potentiating activity of B lymphocyte by increasing the alkaline phosphatase activity by 1.6 fold compared with control at the concentration of 50~500 ${\mu}g/ml$. Fr. Na generated 15.9 ${\mu}M$ of nitric oxide (NO) when cultured with RAW 264.7 at the concentration of 200 ${\mu}g/ml$, while lipopolysaccharide, a positive control, produced 3.7 ${\mu}M$. The Fr. NaCl, Fr. HW and Fr. MeOH increased the secretion of TNF-${\alpha}$, IL-$1{\beta}$, Il-2 and IL-6 by 2.2 times compared with the control group. Fr. Na increased the numbers of peritoneal exudate cells by 4 folds at the concentration of 50mg/kg compared with control. Circulating leukocytes increased by 2.7 folds when Fr. HW from A. brasiliensis was inoculated at the concentration of 50 mg/kg body weight. The hematological and blood chemical analysis of the 3 fractions did not show any difference in blood compositions and enzyme activities compared with the control group (p<0.05). Therefore, the experimental results suggested that crude polysaccharides extracted from A. brasiliensis contain antitumor and immuno-potentiating activities against Sarcoma 180 in ICR mice.

Effects of Dietary Supplementation of Copper-soy Proteinate on the Performance of Laying Hens (Copper-soy Proteinate의 첨가가 채란계의 생산성에 미치는 영향)

  • Lee, Mun-Ku;Kim, Chan-Ho;Shin, Dong-Hun;Jung, Byoung-Yun;Paik, In-Kee
    • Korean Journal of Poultry Science
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    • v.38 no.3
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    • pp.165-171
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    • 2011
  • This study was conducted to investigate the effects of dietary supplementation of copper-soy proteinate (Cu-SP) on the performance of laying hens. A total of 1,000 Hy-Line Brown laying hens of 32 wks old were assigned to one of the following 5 dietary treatments: Control, Cu-SP 50, 100, 150, 200 (50, 100, 150, 200 ppm Cu supplementation as Cu-soy proteinate). Each treatment was replicated 4 times with fifty birds per replication, housed in 2 birds cages. Fifty birds units were arranged according to randomized block design. Feeding trial lasted 5 wks under 16L : 8D lighting regimen. There were no significant differences among treatments in hen-day and hen-house egg production, egg weight, broken & soft egg production. Feed intake and feed conversion rate (FCR) were significantly (P<0.01) lower in Cu-SP treated groups than Control. Eggshell thickness was significantly (P<0.01) higher in Control than Cu-SP 100, 150 and 200. Egg yolk color index was significantly (P<0.01) higher in Cu-SP 200 than other treatments. Egg shell color index was significantly (P<0.05) higher in Cu-SP 150 and Cu-SP 200 than Cu-SP 100. Concentration of copper, iron and zinc of the egg yolk were not significantly influenced by treatment. There were no significant differences in the level of leukocytes and erythrocytes in the chicken blood. The result of this experiment showed that dietary supplementation of Cu-SP at the level of 50ppm of Cu can reduce feed intake and FCR.

Clinical significance of follow-up laboratory tests, performed at 6 weeks after the onset of Kawasaki disease (가와사끼병에서 발병 6주째에 시행하는 추적검사의 임상적 유용성에 관한 고찰)

  • Oh, Il Ji;Moon, Kyung Hee;Hong, Myung Eun;Kim, Yeon Soon;Lee, Chang Woo;Yoon, Hyang Suk
    • Clinical and Experimental Pediatrics
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    • v.49 no.6
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    • pp.672-676
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    • 2006
  • Purpose : Low dose aspirin is used due to its antiplatelet effect for the subacute phase of Kawasaki disease(KD). It is usually used for 6-8 weeks, then various hematologic laboratory tests and follow up echocardiography for evaluating coronary abnormalities are performed. Our review investigated the usefulness of various follow up laboratory tests performed at 6 weeks after the onset of KD. Methods : Two hundred eighty-two children diagnosed and hospitalized with KD were identified by reviewing patient's charts. Cases which were diagnosed between January 1997 and December 2004 were included in this study. We reviewed laboratory data including leukocytes, platelet counts, antistreptolysin O(ASO), erythrocyte sedimentation rate(ESR), C-reactive protein(CRP), aspartate aminotransferase(AST), alanine aminotransferase(ALT), urinalysis, and echocardiograms performed at admission and 6 weeks after the onset of KD. Paired t-test and Fisher's exact test, as well as logistic regression tests, were used for the statistical analysis. Results : At 6th week data, ESR and CRP were still elevated in 35(12.4 percent) and 12(4.3 percent) patients, respectively. Sterile pyuria were all normalized. But, 36 patients(12.8 percent) showed thrombocytosis, 22(7.8 percent) elevated AST, 15(5.3 percent) leukocytosis, and 6(2.1 percent) coronary abnormalities. Coronary abnormalities at the 6th week were only shown in patients with initial abnormalites. Younger age and initial thrombocytosis were risk factors for thrombocytosis at the 6th week. Conclusion : All children with initial coronary abonormalites should have an echocardiogram at 6 weeks after the onset of fever. In view of case-effectiveness, additional echocardiographic studies are justified only if abnormalities are present at admission. ESR, CRP, and urinalysis performed at the 6 weeks after onset of KD is not significant for clinical information of progression. Platelet count should be estimated at 6th week for a judgement of continuous antiplatelet therapy.

Effect of Dietary Supplementation of Cu-methionine Chelate and Cu-soy Proteinate on the Performance, Small Intestinal Microflora and Immune Response in Laying Hens (사료내 Cu-methionine Chelate와 Cu-soy Proteinate가 산란계의 생산성, 소장내 미생물 균총 및 면역체계에 미치는 영향)

  • Paik, I.K.;Kim, C.H.;Park, K.W.
    • Korean Journal of Poultry Science
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    • v.35 no.3
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    • pp.303-311
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    • 2008
  • This study was conducted to investigate the effect of dietary supplementation of Cu-methionine chelate(Cu-Met) and Cu-soy proteinate(Cu-SP) on the performance, small intestinal microflora and immune response in laying hens. A total of 960 Hy-line $Brown^{(R)}$ laying hens of 39 wks old were assigned to one of the following 6 dietary treatment: control(C), antibiotic(Avilamycine 6 ppm), Cu-Met 50 and Cu-Met 100(50 and 100 ppm Cu as Cu-methionine chelate), Cu-SP 50 and Cu-SP 100(50 and 100ppm Cu as Cu-soy proteinate). Each treatment was replicated 4 times with forty birds per replication, housed in 2 birds per cages. Forty birds units were arranged according to randomized block design. Feeding trial lasted 6 wks under 16 hours lighting regimen. Hen-day and hen-house egg production of groups treated with Antibiotic and Cu supplements tended to be higher than the control with significant difference (P<0.05) shown between Cu-Me 100 and control. Egg weight was significantly (P<0.05) heavier in antibiotic and Cu-SP treatments than Cu-Met treatments but they were not significantly different from the control. Eggshell strength, egg shell thickness, egg yolk color and Haugh unit were not significantly different among treatments. There were no significant differences in leukocytes and erythrocytes in the chicken blood. But mean corpuscular hemoglobin value(MCH) was significantly (P<0.05) higher in Cu-SP 100 than antibiotic treatment. The concentrations of serum IgG and IgA were not significantly different among treatments. Copper concentration in the liver tended to increase as the level of copper supplementation increased, that of Cu-SP 100 being significantly (P<0.05) higher than those of the control and antibiotic treatment. Concentrations of iron and zinc of the liver were not significantly influenced by treatments. Populations of Cl. perfringens and Lactobacilli in the small intestinal content were significantly (P<0.05) influenced by treatments. Population of Cl. perfringens decreased and that of Lactobacilli increased in the copper supplemented groups. The result of this experiment showed that Cu-Met and Cu-SP are comparable to antibiotic in improving egg production in laying hens. Birds fed diets supplemented with Cu-SP produced heavier eggs than those fed diets with Cu-Met. There were no significant differences in the performances between 50 ppm and 100 ppm copper supplementation as organic forms.

Hematological Aspects in A Endotoxemic Young Rabbit Model

  • Park, Seok-Cheol;Kwon, Heun-Young;Kim, Jai-Young;Hwang, Soo-Myung;Kim, Tae-Un;Seong, Hee-Kyung;Kim, Yang-Weon;Lee, Won-Jae
    • Biomedical Science Letters
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    • v.8 no.3
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    • pp.115-125
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    • 2002
  • Gram-negative septicemia, which continues to be a serious clinical problem, is one of the major causes of morbidity and mortality in hospitalized patients. Endotoxin of gram-negative bacteria is a pivotal pathogen of sepsis. To understand the effect of endotoxin on hematological aspect and the time course in early childhood, this study was designed with experimental septic model of young rabbits (8 week-old). Rabbits were divided into control (n=7) and endotoxin group (0.50 mg/kg of endotoxin). The endotoxin group was subdivided into six groups by the sampling times: 3, 6, 12, 24, 48 and 72 hr-group (E-G$_{3}$, E-G$_{6}$, E-G$_{12}$, E-G$_{24}$, E-G$_{48}$ and E-G$_{72hrs}$, each n=7). The evaluation of CBC, activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen concentration, coagulation factors and D-dimer were taken from the bloods. The number of leukocytes was lower in E-G$_{3}$ and E-G$_{6hrs}$ (due to pantocytopenia), whereas it was higher in E-G$_{24}$ and E-G$_{48}$ (due to neutrophilia and/or lymphophilia) than in control group (P<0.05). Platelet counts in E-G$_{3}$, E-G$_{6}$, E-G$_{12}$, E-G$_{24}$ and E-G$_{48hrs}$ were lower than those of control group (P<0.05). Normoblast counts in E-G$_{3}$, E-G$_{6}$, E-G$_{12}$, E-G$_{24}$ and E-G$_{48hrs}$ were higher than those of control group (p<0.01). APTT in E-G$_{3}$, E-G$_{6}$, E-G$_{12}$, E-G$_{24}$ and E-G$_{72hrs}$ were longer while PT in E-G$_{3}$, E-G$_{6}$, E-G$_{48}$ and E-G$_{72hrs}$ were higher than those of control group (p<0.05). Fibrinogen concentrations were lower in E-G$_{3}$, E-G$_{6}$ and E-G$_{12}$ but higher in E-G$_{48}$ and E-G$_{72hrs}$ than those of control (p<0.05). Intrinsic coagulation factors (XII, XI, IX, VIII) in all endotoxin groups were significantly lower than those of control group (p<0.05). Extrinsic coagulation factor (X, VII, V, II) were lower in E-G$_{3}$, E-G$_{6}$, E-G$_{12}$ and E-G$_{24hrs}$ whereas they were higher in E-G$_{48}$ and E-G$_{72hrs}$ than in control group (p<0.05). D-dimer concentrations in E-G$_{48}$ and E-G$_{72hrs}$ were higher than those of control group (P<0.001). We concluded that endotoxin led to extensive hematological disturbances including disseminated intravascular coagulation in the young rabbits and that this pathologic condition in the infant and childhood groups will cause the grave results.

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Urine Collection from Disposable Diapers in Infants and Young Children (유소아에서 일회용 기저귀를 이용한 소변 채취 방법)

  • Kim Mi Jeong;Kim Ji Hae;Yim Hyung Eun;Kang Hee;Eun Baik Lin;Yoo Kee Hwan;Hong Young Sook;Lee Joo Won;Kwon Junga
    • Childhood Kidney Diseases
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    • v.9 no.2
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    • pp.128-136
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    • 2005
  • Purpose : Urine collection using a sterile adhesive bag for urinalysis has been used commonly in infants and young children. However, this method has had some drawbacks. So, this study was performed to evaluate the usage of disposable diapers as a substitute for the routine urine collection method. Methods : 60 mL of self voided fresh urine was collected from 99 patients. Half of the urine was poured on the disposable diapers which did not contain absorbent gel beads. After 1 hr, we obtained the urine samples(extracted urine) by compressing the wet diapers using a 50 ml syringe. Routine, microscopic and biochemical analyses were performed on the other half of fresh urine and extracted urine. Then we compared each result by correlative analysis. Results : 198 samples from 99 patients were evaluated. The results of routine urinalysis except WBC and biochemical urinalysis showed a significant correlation between the two groups(P<0.05). The relative coefficients of urine SG, pH, glucose, protein, blood and leukocytes between the two groups were 0.964, 0.938, 0.977, 0.956, 0.931 and 0.738, respectively. Those of urinary sodium, potassium, chloride and creatinine were 0.997, 0.998, 0.995 and 0.998, respectively Microscopic examinations showed lower relative coefficients than the other results, 0.740 for RBC and 0.602 for WBC, but these were still significant(P<0.05). Conclusion : The results of the urine analysis with extracted urine from diapers correlates well with that of fresh urine. This new method is very helpful and can be used as an alternative of urine collection, especially for infants and young children. (J Korean Soc Pediatr Nephrol 2005;9:128-136)

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Effect of Reperfusion after 20 min Ligation of the Left Coronary Artery in Open-chest Bovine Heart: An Ultrastructural Study (재관류가 허혈 심근세포의 미세구조에 미치는 영향 : 재관류 손상에 관한 연구)

  • 이종욱;조대윤;손동섭;양기민;라봉진;김호덕
    • Journal of Chest Surgery
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    • v.31 no.8
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    • pp.739-748
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    • 1998
  • Background: It has been well documented that transient occlusion of the coronary artery causes myocardial ischemia and finally cell death when ischemia is sustained for more than 20 minutes. Extensive studies have revealed that ischemic myocardium cannot recover without reperfusion by adequate restoration of blood flow, however, reperfusion can cause long-lasting cardiac dysfunction and aggravation of structural damage. The author therefore attempted to examine the effect of postischemic reperfusion on myocardial ultrastructure and to determine the rationales for recanalization therapy to salvage ischemic myocardium. Materials and methods: Young Holstein-Friesian cows(130∼140 Kg body weight; n=40) of both sexes, maintained with nutritionally balanced diet and under constant conditions, were used. The left anterior descending coronary artery(LAD) was occluded by ligation with 4-0 silk snare for 20 minutes and recanalized by release of the ligation under continuous intravenous drip anesthesia with sodium pentobarbital(0.15 mg/Kg/min). Drill biopsies of the risk area (antero-lateral wall) were performed at just on reperfusion(5 minutes), 1-, 2-, 3-, 6-, 12-hours after recanalization, and at 1-hour assist(only with mechanical respiration and fluid replacement) after 12-hour recanalization. The materials were subdivided into subepicardial and subendocardial tissues. Tissue samples were examined with a transmission electron microscope (Philips EM 300) at the accelerating voltage of 60 KeV. Results: After a 20-minute ligation of the LAD, myocytes showed slight to moderate degree of ultrastructural changes including subsarcolemmal bleb formation, loss of nuclear matrix, clumping of chromatin and margination, mitochondrial destruction, and contracture of sarcomeres. However, microvascular structures were relatively well preserved. After 1-hour reperfusion, nuclear and mitochondrial matrices reappeared and intravascular plugging by polymorphonuclear leukocytes or platelets was observed. However, nucleoli and intramitochondrial granules reappeared within 3 hours of reperfusion and a large number of myocytes were recovered progressively within 6 hours of reperfusion. Recovery was apparent in the subepicardial myocytes and there were no distinct changes in the ultrastructure except narrowed lumen of the microvessels in the later period of reperfusion. Conclusions: It is likely that the ischemic myocardium could not be salvaged without adequate restoration of coronary flow and that the microvasculature is more resistant to reversible period of ischemia than subendocardium and subepicardium. Therefore, thrombolysis and/or angioplasty may be a rational method of therapy for coronarogenic myocardial ischemia. However, it may take a relatively longer period of time to recover from ischemic insult and reperfusion injury should be considered.

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Histological Changes in Rat Testis by Injection of Hypertonic Saline (고장성 식염수 주사에 의한 흰쥐 정소의 해부학적 변화)

  • Kwak, Byung-Kuk;Lee, Chul-Sang;Lee, Sung-Ho
    • Development and Reproduction
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    • v.14 no.4
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    • pp.281-286
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    • 2010
  • Recent evidence has revealed that the intratesticular injection of hypertonic saline(20%) resulted in a chemically castrated state such as nadir testosterone levels in rats. To confirm the efficacy of this simple saline-injection method further, we investigated the changes in the gross and microscopic anatomy of testis. Our study comprised three groups; intact(control) group, orchidectomy group and saline-injection (experimental) group. Single dose of hypertonic saline (sterilized, $750{\mu}{\ell}/testis$) were directly administered into both testis of adult rats (about 300 g BW). Bilateral orchidectomy was performed at the same day of saline injection. Following 30 days post-injection, reproductive tissues were surgically removed, weighed and fixed for histological examination. The body weights were not changed in both orchidectomy group and saline-injection group when compared to those in intact group. The wet weights of testis were significantly decreased in saline-injection group when compared to those in intact group. The wet weights of epididymis and seminal vesicle and prostate were significantly decreased in orchidectomy group and saline-injection group when compared to those in intact group. Macroscopically, the testes exerted slight atrophy and the tunica albuginea seemed to be intact in saline injection group. Histologically, however, larger parts of testicular tissue underwent necrosis and were barely recognizable after hematoxylin-eosin staining. In the same section, only the opposite part of the injection site was stained showing abnormal state of cell layers mostly fibrosis and infiltrated leukocytes. Sloughing of immature germ cells from the basement membrane along with shedding cells in the intraluminal space was notable in most seminiferous tubules from the saline injected testis. The present study confirmed that the direct injection of hypertonic saline into testis can induce a castration-like, testosterone-depriving effects on accessory sex organs. Our findings suggest that the efficacy of this less expensive and minimally invasive method seems to be almost even with that of conventional orchidectomy and chemical castration, though more in-depth evaluation should be supported.

Histological Observation of Bone Reaction with Relation to Surface Treatment of Titanium (생체합금 Titanium의 표면처리에 따른 골반응의 변화에 대한 조직학적 관찰)

  • Ahn, Myun-Whan;Choi, Yong-Sug;Ahn, Jong-Chul;Seo, Jae-Sung;Lee, Dong-Chul;Jang, Jae-Hyuk
    • Journal of Yeungnam Medical Science
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    • v.13 no.1
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    • pp.116-133
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    • 1996
  • The biocompatibility of the titanium has been estabilished through various experimental studies such as cell culture toxicity test, pyrogen test, mutagen test and others. In order to confirm biocompatibility after fabrication of titanium and to clarify the difference between the bone reaction after insertion of the lathed titanium rods and the bone reaction after insertion of the finished and polished rods, both rods were implanted into the proximal femur of a rabbit. Histologic reactions in the bone were observed according to the ASTM standards at the intervals of 6 weeks, 12 weeks and 26 weeks after implantation. The result were as follows : In 6 weeks after implantation of lathed titanium rods, inflammatory reactions, such as minimal degree infiltration of polymorphonuclear leukocytes and lymphocytes were observed in all cases. This was thought to he caused by surgical trauma. However, inflammatory cell infiltration was not seen after implantation of polished and finished rods in all cases. The cellular infiltration and the histologic reaction of the hone after implantation of lathed group were significantly more pronounced than those after implantation of the finished group. In 12 weeks after implantation of lathed rods, two of four cases revealed a minimal degree of cellular infiltration. No inflammatory cell infiltration was demonstrated after implantation of the finished group. The cellular infiltration and histologic reaction seemed to be more pronounced in the lathed group, but they were not significant statistically. At 26 weeks after implantation of the lathed and finished group, there was no cellular infiltration in both groups. New bone formation was observed up 26 weeks, and no difference between lathed titanium rods and finished titanium rods were apparent. Mild bone necrosis was observed in 1 case out of 11 cases in which lathed titanium rods were implanted. Bone necrosis was not observed in the finished titanium rod group. Fibrosis was observed in both groups, but differences were not significant between the experimental groups. In the lathed titanium rods group and the shorter interval group, inflammatory cell infiltration was significantly higher. Finished titanium rods and longer interval groups had markedly decreased tendences in histologic reaction ratings. As a conclusion, although certificated titanium might be safe to use, difference of biocompatibility were observed depending on the method of surface finish. By identifying biocompatibility as a long-term standardized animal study, we can develop progressed internal fixation device that is safe for human beings.

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The Relationship between Human Neutrophil Elastase and Coronary Arterial Dilatation in Kawasaki Disease (가와사끼병 환아에서의 혈장 및 호중구의 Elastase 활성도와 관상동맥 증대와의 관계)

  • Shim, Jun Yong;Choi, Hee Won;Hong, Ja Hyun;Lee, Jong Kyun;Lee, Hae Yong
    • Clinical and Experimental Pediatrics
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    • v.46 no.9
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    • pp.903-908
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    • 2003
  • Purpose : Kawasaki disease is notorious for coronary arterial complication which is usually developed as a febrile disease in early childhood. Increased polymorphonucleus(PMN) cell levels in acute phases may be associated with the pathophysiology of Kawasaki disease. We studied the relationship between coronary arterial dilatation and elastase activity which was excreted from PMN cell and roles as an important factor for vasculitis. Methods : Ten patients diagnosed with Kawasaki disease in Yonsei University Medical Center were examined between November, 2001 and January, 2002. In addition, 15 patients with other febrile diseases were also examined. Echocardiography was done in patients with Kawasaki disease on the first day of admission and four weeks after the onset of the disease. At each time, venous samples were drawn and separated into plasma and leukocytes. In patients with other febrile disease, samples were drawn on admission. Elastase activities in plasma and neutrophil extracts were measured. Results : The significant increased plasma elastase activity, $6.19{\pm}0.74U/mL$, found in Kawasaki disease patients compared with the other febrile disease patients, $4.86{\pm}1.17U/mL$(P<0.05). And there was no significance between the above two diseases in terms of the elastase activity in neutrophil extracts. The relationship between initial elastase activity and the coronary arterial complication which was shown in subacute phase wasn't significant. Conclusion : Plasma elastase activity was increased in Kawasaki disease significantly, but the initial plasma elastase activity in the acute phase could not reflect the range of coronary arterial complication.