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Multiplex PCR method for environmental monitoring of approved LM cotton events in Korea (국내 승인 LM면화의 자연환경 모니터링을 위한 multiplex PCR 개발)

  • Jo, Beom-Ho;Seol, Min-A;Shin, Su Young;Kim, Il Ryong;Choi, Wonkyun;Eum, Soon-Jae;Song, Hae-Ryong;Lee, Jung Ro
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.91-98
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    • 2016
  • The growth area of living modified (LM) cotton has steadily increased every year, since its first commercialization in 1996. Development of environmental risk assessment tools and techniques for LM cotton is required for ecosystem safety. We therefore developed multiplex PCR assays for simultaneous detection of two (MON15985, MON531) and four (GHB614, LLCOTTON25, MON88913 and MON1445) LM cotton events approved in Korea, with event specific primer pairs. The PCR reactions were optimized by using event specific primers of six LM cottons at various concentrations. The reactions allows amplification of estimated amplicons of MON15985 (214 bp), MON531 (270 bp), GHB614 (119 bp), LLCOTTON25 (164 bp), MON88913 (276 bp), and MON1445 (389 bp) from multiplex PCR reactions. The multiplex PCR assay developed allowed that two annealing steps (15 cycles at $55^{\circ}C$ and 25 cycles at $60^{\circ}C$) were performed for amplification of distinguished two LM cottons, and only one annealing step (50 cycles at $60^{\circ}C$) was necessary for tetraplex PCR. Primer extension step of all PCR reactions was skipped for time-effective amplification. Our methods suggest that two multiplex PCR assays can be cost-effective and a rapid diagnostic tool for environmental LMO monitoring of six LM cottons.

Clinical Significance of Coagulation Screening Tests and Platelet Counts in Children Undergoing Endoscopy (소아에서 내시경 검사의 출혈 위험 예측 인자로서 혈액 응고 검사 및 혈소판 검사의 의의)

  • Lee, Eun-Hye;Yang, Hye-Ran;Ko, Jae-Sung;Seo, Jeong-Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.13 no.1
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    • pp.23-29
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    • 2010
  • Purpose: The aim of this study was to assess the benefits of routine pre-endoscopy coagulation screening tests and platelet counts in Korean children. Methods: Between March 2004 and December 2009, children who underwent gastrointestinal endoscopy for the evaluation of various gastrointestinal symptoms were included. All of the subjects included in the study also underwent routine coagulation screening and platelet count determinations prior to endoscopy and biopsy. The clinical records and laboratory tests were retrospectively reviewed in all patients. Results: One hundred sixty-two of 1,476 (11%) patients who underwent endoscopy had abnormal results on pre-screening coagulation tests. Fourteen patients underwent coagulation factor assays due to abnormal clotting results in consecutive tests or due to clinical evidence of a bleeding tendency. Seven patients were diagnosed with factor XII deficiency, one patient was diagnosed with von Willebrand disease, one patient had von Willebrand disease and factor XII deficiency, and one patient was presumed to have mild hemophilia. The remaining 4 patients had normal results with the factor assays. The results of platelet counts were normal with the exception of 1 patient. No patient had significant bleeding during the endoscopic procedures, despite abnormal pre-endoscopic coagulation tests. Conclusion: Routine coagulation screening tests and platelet counts revealed abnormal results in some patients. Most of the patients with abnormal clotting were shown to have a factor XII deficiency, which had no significant associated bleeding tendencies; the other patients were diagnosed with hemophilia or von Willebrand disease. Therefore, although abnormal pre-endoscopic coagulation is not always related to significant bleeding complications, pre-endoscopic coagulation screening may be useful in some children in predicting the risk of bleeding tendency during endoscopic procedures.

Posttransplantation Lymphoproliferative Disorder after Liver Transplantation in Pediatric Patients: Report from a Single-center Over 21 Years (간 이식 소아에서 발생한 이식 후 림프 증식 질환: 단일 기관에서의 21년 경험)

  • Lee, Jung-Hwa;Ko, Jae-Sung;Seo, Jeong-Kee;Yi, Nam-Joon;Suh, Kyung-Suk;Lee, Kuhn-Uk;Kang, Gyeong-Hoon
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.12 no.2
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    • pp.199-206
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    • 2009
  • Purpose: To analyze the clinical spectrum of posttransplantation lymphoproliferative disorder (PTLD) after liver transplantation in children. Methods: From January 1988 to June 2009, we retrospectively reviewed the medical records of 8 PTLD cases among 148 pediatric patients underwent liver transplantation. The age at transplantation, time of presentation after transplantation, clinical manifestations, histologic diagnosis, results of EBV (Epstein-Barr virus) assessments, managements and outcomes of PTLD were investigated. Results: The prevalence of PTLD in liver transplant pediatric recipients was 5.4% (8 of 148). The mean age of patients was 25.4${\pm}$21.3 months (range 10 to 67 months). Seven of 8 patients (87.5%) underwent liver transplantation before 1 year of age. The common clinical presentations were persistent fever (8 of 8, 100%) and bloody diarrhea (6 of 8, 75%). PTLD was diagnosed with gastrointestinal endoscopic biopsies in five patients and surgical biopsies in three. Histologic findings showed early lesion in three patients, polymorphic in two, and monomorphic in three. Burkitt lymphoma and lymphoblastic lymphoma were found in two of 3 monomorphic patients. Seven of 8 patients were found with EBV-positive. Eight patients were treated with dose reduction of immunosuppressants and infusion of ganciclovir. Rituximab was added to four patients. PTLD were successfully managed in all patients except one who died of sepsis during chemotherapy. Conclusion: Major risk factor of PTLD was to undergo liver transplantation before 1 year of age. Continuous monitoring for EBV viral load and gastrointestinal endoscopic biopsy may be useful to early detection of PTLD.

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Nonalcoholic Fatty Liver Disease in Children with Hypopituitarism (뇌하수체저하증 소아에서 발생한 비알코올성 지방간질환)

  • Yoon, Jung-Min;Ko, Jae-Sung;Seo, Jeong-Kee;Shin, Choong-Ho;Yang, Sei-Won;Moon, Jin-Soo;Yang, Hye-Ran;Chang, Ju-Young
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.13 no.1
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    • pp.51-57
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    • 2010
  • Purpose: It has been reported that children with hypopituitarism have features of metabolic syndrome, including obesity, impaired glucose tolerance, and dyslipidemia. The aim of this study was to investigate the clinical features and liver histology of pediatric non-alcoholic fatty liver disease (NAFLD) associated with hypopituitarism. Methods: We reviewed the clinical data of 11 children diagnosed with NAFLD among patients with hypopituitarism. Results: The mean age at the time of diagnosis of hypopituitarism was 10.4${\pm}$3.2 years, and the mean age at the time of diagnosis of NAFLD was 13.1${\pm}$2.7 years. A craniopharyngioma was the most common cause of pituitary dysfunction. At the time of diagnosis of NAFLD, 9 patients (82%) had a body mass index greater than the 85th percentile, 5 patients (45%) had elevated fasting blood glucose levels, and 9 patients (82%) had hypertriglyceridemia. The mean height SD score was significantly lower at the time of diagnosis of NAFLD than at the time of diagnosis of hypopituitarism. Of the six patients who were biopsied, one had cirrhosis, two had non-alcoholic steatohepatitis (NASH) with bridging fibrosis, two had NASH with mild portal fibrosis, and one had simple steatosis. Conclusion: Children with hypopituitarism are at risk of short stature, obesity, dyslipidemia, and NAFLD. The early diagnosis of NAFLD is important in children with hypopituitarism because advanced fibrosis is common.

Tuberculosis in Infants (영아의 결핵)

  • Kim, Ee-Kyung;Nah, Song-Yi;Park, Jin-Young;Choi, Eun-Hwa;Lee, Hoan-Jong
    • Pediatric Infection and Vaccine
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    • v.5 no.1
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    • pp.69-78
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    • 1998
  • Purpose : The risk of severe tuberculous disease such as meningitis or miliary tuberculosis increases as younger is the child at the time of infection. Therefore, the early diagnosis and prompt treatment is mandatory for infants with tuberculosis. This study was undertaken to describe the epidemiology, clinical and radiographic manifestations, and response to therapy in infants with tuberculous disease. Methods : Medical records of 29 infants with tuberculosis diagnosed at the Seoul National University Children's Hospital from July, 1985, to April, 1997, were reviewed, retrospectively. A case of tuberculosis was confirmed if M. tuberculosis was isolated from any body site or if there was histologic proof of tuberculosis. Otherwise, the diagnoses were individualized considering history of contact with contagious adult case, clinical manifestations, chest X-ray findings, result of a Mantoux test reaction with 5 tuberculin unit of PPD, and the response to therapy. Results : The mean age at diagnosis was $7.00{\pm}2.65$ months (range, 3 to 12 months). Twelve cases had isolated pulmonary diseases, and the rest had pulmonary disease and meningitis, 5 cases; pulmonary disease and cervical lymphadenitis, 3; isolated meningitis, 3; and miliary tuberculosis, 6. Source case was identified in 19 cases, 7 of which were detected with retrograde manner. Twenty seven of 29 were symptomatic at their initial visit. The presenting symptoms were mainly respiratory or neurologic, and respiratory difficulty was accompanied in 7 cases. Physical examination revealed wheezing in 7 cases and decreased breath sounds in 9. Hepatomegaly or hepatosplenomegaly were frequent. Chest radiographs showed lung parenchymal disease with hilar lymphadenopathy in 18 cases, and focal or generalized emphysematous change in 7 cases. Conclusion : Most of the infants with tuberculosis are symptomatic at diagnosis, and many of infants with intrathoracic tuberculosis presented with symptoms of bronchial obstruction. When tuberculosis is suspected in an infant, the adult source case should be vigorously investigated to aid in diagnosis and for the prevention of further transmission of tuberculous disease. Almost half of infant tuberculosis are preventable if prophylaxis were given when adult cases were diagnosed.

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Ventilator-Associated Pneumonia in Neonatal Intensive Care Unit : Clinical Manifestations, Ddiagnostic Availability of Endotracheal Tip Culture (신생아 집중치료실에서 인공 환기요법 관련 폐렴의 임상양상과 기관내 삽관의 유용성에 관한 연구)

  • Kim, Nam Young;Sung, Tae Jung;Shin, Seon Hee;Kim, Sung Koo;Lee, Kon Hee;Yoon, Hae Sun
    • Pediatric Infection and Vaccine
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    • v.12 no.1
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    • pp.67-74
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    • 2005
  • Purpose : The incidence of mortality associated with respiratory difficulties is decreasing nowadays contributed to the development of neonatology. However, complications associated with mechanical ventilator are increasing. This study is to determine clinical manifestations, diagnositc availability of the endotracheal tip culture in patients with Ventilator-Associated Pneumonia(VAP) in neonatal intensive care unit(NICU). Methods : A retrospective analysis of 50 neonates who were admitted to the NICU of Kangnam Sacred Heart Hospital and had given mechanical ventilator from 1 January 2000 to 30 June 2003. VAP group defined as neonates who had pneumonia with mechanical ventilation longer than 48 hours. They were classified into VAP group(n=13) and control group (n=37) and the prevalence, microorganisms cultured from the endotracheal tube tip and risk factors were investigated. Results : The prevalence of VAP was 26.0%(n=13) and the most dominant microorganism cultured in our NICU was methicillin-resistant coagulase negative staphylococcus(MR-CNS) in 4 cases. Other microorganisms were Pseudomonas, Enterobacter, methicillin-resistant Staphylococcus aureus(MRSA) and Klebsiella. Gestational age, birth weight, Apgar score, respiratory distress syndrome, retinopathy of prematurity, bronchopulmonary dysplasia, sepsis, renal failure, pulmonary hemorrhage, pneumothorax were not different significantly between two groups except intraventricular hemorrhage(P<0.001) and patent ductus arteriosus(P<0.05). Duration of hospital stay and mortality rate were also not different significantly. Conclusion : VAP occurred at a significant rate among mechanically ventilated NICU patients. Despite of limitation of encotracheal tip culture, the most common microorganism was MR-CNS. We should be aware of occurrence of VAP in NICU neonate who were with mechanical ventilator and should treat with great care.

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The change of validity of blood zinc protoporphyrin test by different cut-off level in lead workers (연취급 근로자들의 혈중 ZPP 농도 선별기준에 따른 정확도의 변화)

  • Kim, Yong-Bae;Ahn, Hyun-Cheol;HwangBo, Young;Lee, Gap-Soo;Lee, Sung-Soo;Ahn, Kyu-Dong;Lee, Byung-Kook
    • Journal of Preventive Medicine and Public Health
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    • v.30 no.4 s.59
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    • pp.741-751
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    • 1997
  • Measurement of blood lead (PbB) and blood zinc protoporphyrin (ZPP) are most common biological indices to identify the individual at risk for excess or the health sequences by lead exposure. Because PbB is known most important and reliable index of lead exposure, PbB is often regarded as a gold standard to detect lead exposure. But in Korea PbB is a secondary test item of detailed health check-up with positive finding of screening test in most occasion. Our lead standard requires all lead workers to take annual heath-check twice a year for investigation of their health effect due to lead exposure. Blood ZPP is one of most important index to detect high lead absorption in lead workers as a screening test. Measurement of blood ZPP is known ,well to correlate with PbB in steady state of exposure in most lead workers and is often used as a primary screening test to detect high lead absorption of lead workers with the advantage of simplicity, easiness, portability and low cost. The current cut-off criteria of blood ZPP for further detailed health check-up is $100{\mu}g/d\ell$ which is supposed to match the level of $40{\mu}g/d\ell$ of PbB according to our standard. Authors tried to investigate the validity of current criteria of cut-off level $(100{\mu}g/d\ell)$ of blood ZPP and possible another better cut-off level of it to detect the lead workers whose PbB level over $40{\mu}g/d\ell$. The subjects in our study were 212 male workers in three small scale storage battery industries. Blood ZPP, PbB and hemoglobin (Hb) were selected as the indices of lead exposure. The results were as follows. 1. The mean of blood ZPP, PbB and Hb in lead workers were $79.5{\pm}46.7{\mu}g/d\ell,\;38.7{\pm}15.1{\mu}g/d\ell,\;and\;14.8{\pm}1.2g/d\ell$, respectively. There were significant differences in blood ZPP, PbB and Hb by industry (P<0.01). 2. The percents of lead workers whose blood ZPP were above $100{\mu}g/d\ell$ in the group of work duration below 1, 1-4, 5-9 and above 10 years were 8.6%, 17.2%, 47.6%, and 50.0%, respectively. The percents of lead workers whose PbB were above $40{\mu}g/d\ell$ in those were 31.4%, 40.4%, 71.4%, and 86.4%, respectively. 3. The percents of lead workers whose PbB were below $40{\mu}g/d\ell$, $40-59{\mu}g/d\ell$ and above $60{\mu}g/d\ell$ were 54.7%, 34.9% and 10.4%, respectively. Those of lead workers whose blood ZPP were below $100{\mu}g/d\ell$, $100-149{\mu}g/d\ell$ and above $150{\mu}g/d\ell$ were 79.2%, 13.7% and 7.1%, respectively. 4. Simple linear regression of PbB on blood ZPP was statistically significant (P<0.01) and as PbB was $40{\mu}g/d\ell$, blood ZPP was $82.1{\mu}g/d\ell$. 5. While the highest sensitivity and specificity of blood ZPP test to detect lead workers with PbB eve. $40{\mu}g/d\ell$ were observed in the cut-off level of $50{\mu}g/d\ell$ and $100{\mu}g/d\ell$ of blood ZPP, respectively, the highest validity (sensitivity+specificity) of blood ZPP to detect lead workers with PbB over $40{\mu}g/d\ell$ was observed in the cut-off level of around $70{\mu}g/d\ell$ of blood ZPP. But even with optimal cut-off level of around $70{\mu}g/d\ell$ of blood ZPP, still 25.0% of false negative and 20.7% false positive lead workers were found. As the result of this study, it was suggested that reconsideration of current blood ZPP cut-off of our lead standard from $100{\mu}g/d\ell$ to somewhat lower level such as around $70{\mu}g/d\ell$ and the inclusion of PbB measurement as a primary screening test for lead workers was highly recommended for the effective prevention of lead workers.

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BTEX Exposure and its Health Effects in Pregnant Women Following the Hebei Spirit Oil Spill (허베이 스피릿호 유류유출사고 지역 산모의 BTEX 노출과 건강영향)

  • Kim, Byung-Mi;Park, Eun-Kyo;LeeAn, So-Young;Ha, Mi-Na;Kim, Eun-Jung;Kwon, Ho-Jang;Hong, Yun-Chul;Jeong, Woo-Chul;Hur, Jong-Il;Cheong, Hae-Kwan;Yi, Jong-Heop;Kim, Jong-Ho;Lee, Bo-Eun;Seo, Ju-Hee;Chang, Moon-Hee;Ha, Eun-Hee
    • Journal of Preventive Medicine and Public Health
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    • v.42 no.2
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    • pp.96-103
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    • 2009
  • Objectives : We evaluated the health effects of exposure to BTEX(Benzene, Toluene, Ethylbenzene, m,p-Xylene, o-Xylene) in the Taean area after the Hebei Spirit oil spill. Methods : We used a questionnaire survey to look for health effects among 80 pregnant women 2 to 3 months following the Hebei Spirit oil spill. Their BTEX exposures were estimated using the CALPUFF method. We then used a multiple logistic regression analysis to evaluate the effects of BTEX exposure on the women s health effets. Results : Pregnant women who lived near the accident site reported more symptoms of eye irritation and headache than those who lived farther from the site. There was a trend of decreasing symptoms with an increase in distance from the spill site. Pregnant women exposed to higher ambient cumulative levels of Xylene were significantly more likely to report symptoms of the skin(OR 8.01 95% CI=1.74-36.76) in the first day after the accident and significantly more likely to report abdominal pain(OR 3.86 95% CI=1.02-14.59 for Ethylbenzene, OR 6.70 95% CI=1.82-24.62 for Xylene) during the 1st through 4th days following the accident. Conclusions : This study suggests that exposure to BTEX from an oil spill is correlated with an increased risk of health effects among pregnant women. This implies the need to take proper measures, including the development of a national policy for environmental health emergencies and a plan for studying the short- and long-term chronic health effects associated with such spills.

Eosinophilic Peritonitis (EP) Complicated with Continuous Ambulatory Peritoneal Dialysis (CAPD) (지속적 외래 복막투석 직후 발생하는 비감염성 호산구성 복막염)

  • Paek Kyung-Hoon;Jeon Yun-Ae;Min Jae-Hong;Park Kyung-Mi;Kim Jung-Su;Ha Il-Soo;Cheong Hae-Il;Choi Yong;Ko Kwang-Wook
    • Childhood Kidney Diseases
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    • v.1 no.2
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    • pp.117-122
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    • 1997
  • Purpose : To clarify the clinical findings, laboratory findings and disease course of EP complicated with CAPD and to find out possible predisposing factors to EP. Methods : The medical records of 34 children who underwent CAPD at our hospital between Jan. '94 and Dec. '96 were retrospectively reviewed. The clinical features and laboratory findings of EP were analyzed, and several parameters were evaluated as predisposing factors of EP. Results : EP developed in 7(21%) out of 34 patients. The major symptom of EP was turbid peritoneal fluid without fever, abdominal pain or disturbance of drainage in all cases. The microbiologic culture studies of the peritoneal fluid resulted negative in all cases. Patients with peripheral blood eosinophilia before insertion of CAPD catheter had higher risk of EP than those without eosinophilia (P=0.002). And peripheral blood eosinophilia, noted after insertion of hemodialysis catheter in cases with previous hemodialysis before CAPD, showed significant correlation with the occurrence of EP (P=0.016), too. However, there was no significant correlation between peripheral blood eosinophilia noted after insertion of CAPD cathter and the occurrence of EP. Identification of eosinophils in peritoneal fluids was more accurate with cytospin analysis. Conclusions : An early and accurate diagnosis of EP in patients with CAPD can prevent unnecessary treatment of antibiotics. Peripheral blood eosinophilia before insertion of CAPD catheter is one of the predisposing factors of EP. And, cytospin analysis of peritoneal fluid is an accurate method for diagnosis of EP.

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Outcome and Prognosis in Critically III Children Receiving Continuous Renal Replacement Therapy (소아 중환자에서 지속적 신대체요법의 치료 결과와 예후)

  • Park, Kwang-Sik;Son, Ki-Young;Hwang, You-Sik;Kim, Joung-A;Cheung, Il-Chun;Shin, Jae-Il;Park, Ji-Min;Ahn, Sun-Young;Lyu, Chuhl-Joo;Lee, Jae-Seung
    • Childhood Kidney Diseases
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    • v.11 no.2
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    • pp.247-254
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    • 2007
  • Purpose : Continuous renal replacement therapy(CRRT) has been the first choice for the treatment of acute renal failure in critically ill children not only in western countries but also in Korea. However, there are very few studies that have analyzed the outcome and prognosis of this modality in Korean children. We performed this study to evaluate the factors associated with the outcome and prognosis of patients treated with CRRT. Methods : We retrospectively reviewed the medical records of 32 children who had received CRRT at Severance hospital from 2003 to 2006. The mean age was 7.5 years(range 4 days-16 years) and the mean body weight was 25.8 kg (range 3.2-63 kg). Results : Eleven(34.4%) of the 32 patients survived. Bone marrow transplantation and malignancy were the most common causes of death and underlying disease leading to the need for CRRT Mean patient weight, age, duration of CRRT, number of organ failures, urine output, estimated glomerular filtration rate(eGFR), C-reactive protein, and blood urea level did not differ significantly between survivors and nonsurvivors. (1) Pediatric risk of mortality(PRISM) III score at CRRT initiation($9.8{\pm}5.3$ vs. $26.7{\pm}7.6$, P<0.0001), (2) maximum pressor number ($2.1{\pm}1.2$ vs. $3.0{\pm}1.0$, P=0.038), and (3) the degree of fluid overload($5.2{\pm}6.0$ vs. $15.0{\pm}8.9$, P=0.002) were significantly lower in survivers than in nonsurvivors. Multivariate analysis revealed that fluid overload was the only independent factor reducing survival rate. Conclusion : CRRT was successfully applied to the treatment of acute renal failure in a wide range of critically ill children. To improve survival, we suggest the early initiation of CRRT to prevent the systemic worsening and progression of fluid overload in critically ill children with acute renal failure. (J Korean Soc Pediatr Nephrol 2007;11:247-254)

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