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Characterization of MACS Isolated Cells from Differentiated Human ES Cells (인간 배아줄기세포로부터 분화된 세포에서 MACS 방법을 이용하여 분리한 세포의 특성에 대한 연구)

  • Cho, Jae Won;Lim, Chun Kyu;Shin, Mi Ra;Bang, Kyoung Hee;Koong, Mi Kyoung;Jun, Jin Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.3
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    • pp.171-178
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    • 2006
  • Objective: Human embryonic stem (ES) cells have a great potential in regenerative medicine and tissue engineering. The human ES cells could be differentiated into specific cell types by treatments of growth factors and alterations of gene expressions. However, the efficacy of guided differentiation and isolation of specific cells are still low. In this study, we characterized isolated cells from differentiated human ES cells by magnetic activated cell sorting (MACS) system using specific antibodies to cell surface markers. Methods: The undifferentiated hES cells (Miz-hESC4) were sub-cultured by mechanical isolation of colonies and embryoid bodies were spontaneously differentiated with DMEM containing 10% FBS for 2 weeks. The differentiated cells were isolated to positive and negative cells with MACS system using CD34, human epithelial antigen (HEA) and human fibroblast (HFB) antibodies, respectively. Observation of morphological changes and analysis of marker genes expression were performed during further culture of MACS isolated cells for 4 weeks. Results: Morphology of the CD34 positive cells was firstly round, and then it was changed to small polygonal shape after further culture. The HEA positive cells showed large polygonal, and the HFB positive spindle shape. In RT-PCR analysis of marker genes, the CD34 and HFB positive cells expressed endodermal and mesodermal genes, and HEA positive cells expressed ectodermal genes such as NESTIN and NF68KD. The marker genes expression pattern of CD34 positive cells changed during the extension of culture time. Conclusion: Our results showed the possibility of successful isolation of specific cells by MACS system from undirected differentiated human ES cells. Thus, MACS system and marker antibodies for specific cell types might be useful for guided differentiation and isolation of specific cells from human ES cells.

Correlations and Seasonal Variations of Marine Viral Abundances, Bacterial Abundances and Concentration of Chlorophyll-$\alpha$ in Gwangyang Bay (광양만내 해양 바이러스에 대한 엽록소$\alpha$와 박테리아 개체량의 계절적 변이와 상관관계)

  • Choi, Eun-Seok;Kim, So-Jung;Oh, Ro-Ra;Yun, Hee-Young;Shin, Kyung-Soon;Chang, Man;Lee, Sukchan;Lee, Sang-Seob;Lee. Taek-Kyun
    • Korean Journal of Environmental Biology
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    • v.22
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    • pp.86-92
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    • 2004
  • The marine viral density in the Gwangyang Bay was abundant about 2.0${\times}$10$^{8}$ particles ml$^{-1}$ . For each season, viral abundances were recorded from 9.0${\times}$10$^{8}$ particles ml$^{-1}$ in summer to 0.7${\times}$10$^{6}$ particles ml$^{-1}$ in winter. The spatial distributions of the viral, bacterial and phytoplankton biomass in the Gwangyang Bay were mostly highey in closed estuarine system (Station 2, 5, 10, 12, 16, 20) than open ocean system (Station 28, 38, 42, 46, 51), And the othey closed estuarine system (Station 22, 26, 32, 34) indicated higher viral abundances, lower bacterial and plankton biomass than open oceanic system. In depths of some stations, the bacterial abundances exceeded a hundred fold than viral abundances. Seasonal abundances of marine viruses and their host systems were dynamically changed, and their seasonal variations were closely correlated. In summer, viral and bacterial abundances were increased, and phytoplankton chlorophyll $\alpha$ concentrations were maintained in average values. In winter, viral and bacterial abundances were dramatically decreased, and chlorophyll a concentrations were decreased, but, immediately increased. The viral abundances were peaked in August 2001, and bacteyial abundance, in August 2001 and June 2002, while chlorophyll a concentrations were peaked in April. 2002. In total host and viral abundances, it was seemed that their pools were maintained to steady-states by viral mortality, and viral abundance maintained steady-states. In our assessments, this report is a unique research about marine viral ecology of the Gwangyang Bay in Korea.

Design Strategies and Processes through the Concept of Resilience (리질리언스 개념을 통해서 본 설계 전략과 과정)

  • Choi, Hyeyoung;Seo, Young-Ai
    • Journal of the Korean Institute of Landscape Architecture
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    • v.46 no.5
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    • pp.44-58
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    • 2018
  • Cities face new challenges not only in natural disasters by climate change but also in social and economic fluctuations. With the existing simple reconstruction method, it is difficult to solve the overall problems that a city or region may face. As a new approach to cope with various changes, the concept of resilience is emerging. Resilience is also one of the themes of recent major urban design projects. Design with the concept of resilience is a new strategy that can deal with various changes of urban space, rather than a temporary trend. The purpose of this paper is to explore the design method by analyzing cases where the concept of resilience is employed. We aim to examine what kind of design strategies are needed for the resilience design and how this design process differ in character, as compared to general design projects. Cases for this study include the "Rebuild by Design" competition held in 2013 and the "Resilient by Design/Bay Area Challenge" competition held in 2017. This paper consists of literature reviews and case studies. The latter is divided into two aspects: content analysis based on the theory of resilience and characteristics of the design process. Cases are analyzed through literature reviews and process characteristics of resilience design in response to the general design process. The main categories for urban resilience used as the framework for analysis include: Urban Infrastructure, Social Dynamics, Economic Dynamics, Health and Wellbeing, Governance Networks, and Planning and Institutions. As a result, the aspects of resilience concepts considered and design strategies undertaken by each team were identified. Each team tried to connect all 6 categories to their design strategies, placing special value on the role of governance, a system that enables collaborative design and project persistency. In terms of the design process, the following characteristics were found: planning the whole project process in the pre-project phase, analyzing predictable socioeconomic risk factors in addition to physical vulnerabilities, aiming for landscape-oriented integrated design, and sustainable implementation strategies with specific operations and budget plans. This paper is meaningful to connect the concept of resilience, which has been discussed in various articles, to design strategy, and to explore the possibility of constructing a practical methodology by deriving the characteristics of the resilience design process. It remains a future task to research design strategies that apply the concept of resilience to various types of urban spaces, in addition to areas that are vulnerable to disasters.

Effects of electromagnetic stimulation on neurogenesis and neuronal proliferation in rat hippocampal slice culture (실험 쥐 해마조직배양에서 전자기 자극이 신경조직발생 및 증식에 미치는 영향)

  • Kim, Deok-Soo;Choi, Eung Sang;Chae, Soo Ahn
    • Clinical and Experimental Pediatrics
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    • v.49 no.5
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    • pp.558-564
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    • 2006
  • Purpose : Transcranial electromagnetic stimulation(TMS) is a noninvasive method which stimulates the central nervous system through pulsed magnetic fields without direct effect on the neurons. Although the neurobiologic mechanisms of magnetic stimulation are unknown, the effects on the brain are variable according to the diverse stimulation protocols. This study aims to observe the effect of the magnetic stimulation with two different stimulation methods on the cultured hippocampal slices. Methods : We obtained brains from 8-days-old Spague-Dawley rats and dissected the hippocampal tissue under the microscope. Then we chopped the tissue into 450 µm thickness slices and cultured the hippocampal tissue by Stoppini's method. We divided the inserts, which contained five healthy cultured hippocampal slices respectively, into magnetic stimulation groups and a control group. To compare the different effects according to the frequency of magnetic stimulation, stimulation was done every three days from five days in vitro at 0.67 Hz in the low stimulation group and at 50 Hz in the high stimulation group. After N-methyl-D-aspartate exposure to the hippocampal slices at 14 days in vitro, magnetic stimulation was done every three days in one and was not done in another group. To evaluate the neuronal activity after magnetic stimulation, the $NeuN/{\beta}$-actin ratio was calculated after western blotting in each group. Results : The expression of NeuN in the magnetic stimulation group was stronger than that of the control group, especially in the high frequency stimulation group. After N-methyl-D-aspartate exposure to hippocampal slices, the expression of NeuN in the magnetic stimulation group was similar to that of the control group, whereas the expression in the magnetic non-stimulation group was lower than that of the control group. Conclusion : We suggest that magnetic stimulation increases the neuronal activity in cultured hippocamal slices, in proportion to the stimulating frequency, and has a neuroprotective effect on neuronal damage.

Early and mid-term results of pulmonary valve reconstruction in surgical repair of tetralogy of Fallot; comparison with other techniques of right ventricular outflow reconstruction (활로사징 완전교정술에서 폐동맥 판막 재건술의 조기 및 중기 결과; 우심실 유출로 재건 방법에 따른 비교)

  • Wang, Sheng Wen;Lee, Young Seok;Kim, Si Ho;Kim, Tae Hong;Ban, Ji Eun;Lee, Hyoung Doo;Chang, Yun Hee;Sung, Si Chan
    • Clinical and Experimental Pediatrics
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    • v.49 no.6
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    • pp.635-642
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    • 2006
  • Purpose : The purpose of this study is to determine whether the new pulmonary valve reconstruction technique prevents short-term postoperative pulmonary regurgitation and improves early and mid-term clinical outcome. Methods : We reviewed postoperative echocardiographic variables and chest X-ray films from 31 patients who had undergone valve reconstruction(pulmonary valve reconstruction group : PVR) for the repair of TOF between April 2000 and August 2004. We compared the clinical data of these patients with those from 47 patients who had right ventricular outflow tract reconstruction with a monocusp valve(monocusp ventricular outflow patch group : MVOP) and 22 patients who had a transannular patch repair without a monocusp valve(transannular patch group : TAP). Results : In the PVR group, 25 patients(81 percent) had trivial or mild pulmonary regurgitation in their early post operative echocardiogram. Only 12 patients(26 percent) in the MVOP group had mild pulmonary regurgitation; and no patient in the TAP group had it. Pulmonary valve function was good in 96 percent of the PVR group, 36 percent of the MVOP group, and none in the TAP group in early post-operative echocardiogram. Follow-up echocardiogram(1, 2, 3, 4 years later) of the MVOP and TAP groups showed moderate pulmonary regurgitation and severely decreased valve function in almost all cases. However, in the PVR group 54 percent(16/28), 50 percent(14/28), 37 percent(9/24), and 31 percent(5/16) of the patients had trivial or mild pulmonary regurgitation 1, 2, 3 and 4 years after operation, respectively. The valve function remained good in 80 percent(24/30), 64 percent(18/28), 57 percent(12/21), and 31 percent(5/16) of the patients 1, 2, 3 and 4 years after operation respectively. Conclusion : Pulmonary valve reconstruction is effective in reducing pulmonary regurgitation and right ventricular dilatation in the repair of TOF, even though regurgitation increases with time. Further study is needed to determine long-term results.

Association study of polymorphism in leukotriene C4 synthase and cysteinyl leukotriene receptor 1 genes with phenotype of asthma and clinical parameters in Korean children (Leukotrienes C4 synthase와 cysteinyl leukotriene receptor 1 유전자 다형성과 한국 소아 천식 표현형 및 임상 지표와의 연관성 연구)

  • Shim, Jung Yeon;Kim, Byung-Joo;Song, Young Hwa;Kang, Mi-Jin;Lee, So-Yeon;Kim, Hyo-Bin;Yu, Jinho;Hong, Soo-Jong
    • Clinical and Experimental Pediatrics
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    • v.52 no.6
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    • pp.680-688
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    • 2009
  • Purpose : Cysteinyl leukotrienes are important proinflammatory mediators in asthma. Recently, it was suggested that a promoter polymorphism in the genes encoding for leukotriene C4 synthase (LTC4S), a key enzyme in the leukotriene synthetic pathway, and cysteinyl leukotriene receptor 1 (CysLTR1) might be associated with aspirin-intolerant asthma. We investigated whether polymorphisms in LTC4S and CysLTR1 genes or their interactions were associated with the asthma phenotype, lung function, or bronchial hyperreactivity (BHR) in Korean children. Methods : A total of 856 asthmatic children and 254 non-asthmatic controls were enrolled; a skin prick test, lung function test and bronchial provocation test were performed. Of those enrolled, 395 children underwent exercise challenge tests. The LTC4S A(-444)C and CysLTR1 T(+927)C were genotyped by polymerase chain reaction-restriction fragment length polymorphism analysis. Results : Of those enrolled, 699 children were classified as having atopic asthma and 277 children, as having exercise-induced asthma (EIA). LTC4S and CysLTR1 polymorphisms were not associated with atopic asthma, EIA, or asthma per se. Lung function and BHR were not significantly different between the wild type (AA or TT) and the variant (AC+CC or TC+CC) genotypes in asthmatics, atopic asthmatics, and EIA (+) asthmatics, while total eosinophil counts were higher in the variant type of LTC4S than in the wild type in atopic asthmatics. There were no associations between the gene-gene interactions of LTC4S and CysLTR1 genotypes and the asthma phenotypes. Conclusion : LTC4S A(-444)C and CysLTR1 T(+927)C polymorphisms and their gene-gene interactions are not associated with asthma phenotype, lung function, or BHR in Korean children.

Neuroprotective effects of resveratrol via anti-apoptosis on hypoxic-ischemic brain injury in neonatal rats (신생 백서의 저 산소 허혈 뇌손상에서 항세포사멸사를 통한 resveratrol의 신경보호 효과)

  • Shin, Jin Young;Seo, Min Ae;Choi, Eun Jin;Kim, Jin Kyung;Seo, Eok Su;Lee, Jun Hwa;Chung, Hai Lee;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1102-1111
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    • 2008
  • Purpose : Resveratrol, extracted from red wine and grapes, has an anti-cancer effect, an antiinflammatory effect, and an antioxidative effect mainly in heart disease and also has neuroprotective effects in the adult animal model. No studies for neuroprotective effects during the neonatal periods have been reported. Therefore, we studied the neuroprotective effect of resveratrol on hypoxic-ischemic brain damage in neonatal rats via anti-apoptosis. Methods : Embryonic cortical neuronal cell culture of rat brain was performed using pregnant Sprague-Dawley (SD) rats at 18 days of gestation (E18) for the in vitro approach. We injured the cells with hypoxia and administered resveratrol (1, 10, and $30{\mu}g/mL$) to the cells at 30 minutes before hypoxic insults. In addition, unilateral carotid artery ligation with hypoxia was induced in 7-day-old neonatal rats for the in vivo approach. We injected resveratrol (30 mg/kg) intraperitoneally into animal models. Real-time PCR and Western blotting were performed to identify the neuroprotective effects of resveratrol through anti-apoptosis. Results : In the in vitro approach of hypoxia, the expression of Bax, caspase-3, and the ratio of Bax/Bcl-2, indicators of the level of apoptosis, were significantly increased in the hypoxia group compared to the normoxia group. In the case of the resveratrol-treated group, expression was significantly decreased compared to the hypoxia group. And the results in the in vivo approach were the same as in the in vitro approach. Conclusion : The present study demonstrates that resveratrol plays neuroprotective role in hypoxic-ischemic brain damage during neonatal periods through the mechanism of anti-apoptosis.

A case of simultaneously identified glycogen storage disease and mucopolysaccharidosis (당원병과 뮤코다당체침착증이 동시에 발견된 증례 1예)

  • Lee, Ju Young;Shim, Jeong Ok;Yang, Hye Ran;Chang, Ju Young;Shin, Choong Ho;Ko, Jae Sung;Seo, Jeong Kee;Kim, Woo Sun;Kang, Gyeong Hoon;Song, Jeong Han;Kim, Jong Won
    • Clinical and Experimental Pediatrics
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    • v.51 no.6
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    • pp.650-654
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    • 2008
  • Glycogen storage disease (GSD) and mucopolysaccharidosis (MPS) are both independently inherited disorders. GSD is a member of a group of genetic disorders involving enzymes responsible for the synthesis and degradation of glycogen. GSD leads to abnormal tissue concentrations of glycogen, primarily in the liver, muscle, or both. MPS is a member of a group of inherited lysosomal storage diseases, which result from a deficiency in specific enzymatic activities and the accumulation of partially degraded acid mucopolysaccharides. A case of a 16-month-old boy who presented with hepatomegaly is reported. The liver was four finger-breadth-palpable. A laboratory study showed slightly increased serum AST and ALT levels. The liver biopsy showed microscopic features compatible with GSD. The liver glycogen content was 9.3% which was increased in comparison with the reference limit, but the glucose-6-phosphatase activity was within the normal limit. These findings suggested GSD other than type I. Bony abnormalities on skeletal radiographs, including an anterior beak and hook-shaped vertebrae, were seen. The mucopolysaccharide concentration in the urine was increased and the plasma iduronate sulfatase activity was low, which fulfilled the diagnosis criteria for Hunter syndrome (MPS type II). To the best of the authors' knowledge, this is the first case of GSD and Hunter syndrome being identified at the same time.

Feasibility Test on Automatic Control of Soil Water Potential Using a Portable Irrigation Controller with an Electrical Resistance-based Watermark Sensor (전기저항식 워터마크센서기반 소형 관수장치의 토양 수분퍼텐셜 자동제어 효용성 평가)

  • Kim, Hak-Jin;Roh, Mi-Young;Lee, Dong-Hoon;Jeon, Sang-Ho;Hur, Seung-Oh;Choi, Jin-Yong;Chung, Sun-Ok;Rhee, Joong-Yong
    • Journal of Bio-Environment Control
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    • v.20 no.2
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    • pp.93-100
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    • 2011
  • Maintenance of adequate soil water potential during the period of crop growth is necessary to support optimum plant growth and yields. A better understanding of soil water movement within and below the rooting zone can facilitate optimal irrigation scheduling aimed at minimizing the adverse effects of water stress on crop growth and development and the leaching of water below the root zone which can have adverse environmental effects. The objective of this study was to evaluate the feasibility of using a portable irrigation controller with an Watermark sensor for the cultivation of drip-irrigated vegetable crops in a greenhouse. The control capability of the irrigation controller for a soil water potential of -20 kPa was evaluated under summer conditions by cultivating 45-day-old tomato plants grown in three differently textured soils (sandy loam, loam, and loamy sands). Water contents through each soil profile were continuously monitored using three Sentek probes, each consisting of three capacitance sensors at 10, 20, and 30 cm depths. Even though a repeatable cycling of soil water potential occurred for the potential treatment, the lower limit of the Watermark (about 0 kPa) obtained in this study presented a limitation of using the Watermark sensor for optimal irrigation of tomato plants where -20 kPa was used as a point for triggering irrigations. This problem might be related to the slow response time and inadequate soil-sensor interface of the Watermark sensor as compared to a porous and ceramic cup-based tensiometer with a sensitive pressure transducer. In addition, the irrigation time of 50 to 60 min at each of the irrigation operation gave a rapid drop of the potential to zero, resulting in over irrigation of tomatoes. There were differences in water content among the three different soil types under the variable rate irrigation, showing a range of water contents of 16 to 24%, 17 to 28%, and 24 to 32% for loamy sand, sandy loam, and loam soils, respectively. The greatest rate increase in water content was observed in the top of 10 cm depth of sandy loam soil within almost 60 min from the start of irrigation.

The Characteristics of Membranoproliferative Glomerulonephritis I Detected from School Urine Screening (학교 집단 소변 검사로 발견 된 막증식성 사구체신염 I형의 특성)

  • Choi, Jung-Youn;Park, Mi-Young;Lee, Yong-Jik;Ha, Il-Soo;Cheong, Hae-Il;Choi, Yong;Park, Young-Seo;Han, Hye-Won;Jin, Dong-Kyu;Chung, Woo-Yeong;Kim, Kee-Hyuck;Yoo, Kee-Hwan;Park, Yong-Hoon
    • Childhood Kidney Diseases
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    • v.10 no.2
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    • pp.152-161
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    • 2006
  • Purpose : In Korea, the school urine screening program is a useful tool for screening urine abnormalities. It is particularly useful in early detection of membranoproliferative glomerulonephritis(MPGN) I, which frequently progresses to chronic renal failure. In this study, we studied the medical history, laboratory findings, and histologic findings of MPGN to gain helpful information on early detection and treatment. Methods : The subjects were 19 children, who were diagnosed with MPGN from kidney biopsies that were performed in ten nationwide university hospitals because of abnormal urine findings from school urine screening programs conducted from July 1999 to April 2004. We divided the patients into 2 groups, a nephrotic range proteinuria group(n=8) and a non-nephrotic proteinuria group(n=11), and retrospectively analyzed the clinical features, laboratory findings, histologic findings, treatment, and clinical course. Results : The mean age at the first abnormal urinalysis was $10.6{\pm}2.2$ years in the nephrotic proteinuria group and $9.6{\pm}3.2$ years in the non-nephrotic proteinuria group. The mean age at the time of kidney biopsy was $11.3{\pm}2.3$ years in the nephrotic range proteinuria group and $10.4{\pm}3.2$ years in the non-nephrotic proteinuria group respectively. There was no significant difference in the mean age and sex between the two groups. In the nephrotic proteinuria group, 6 children had a low plasma C3 level and in the non-nephrotic proteinuria group, 8 children had a low plasma C3 level, but there was no significant difference between the 2 groups. There was no significant difference in the laboratory test results(including WBC count, RBC count, platelet count and other serologic tests) between the 2 groups except for 24 hour urine protein secretion. There was no difference between the 2 groups with regard to the acute and chronic changes in the glomerulus on light microscopic findings, IgG, IgA, Ig M, C1q, C3, C4, fibrogen deposition on immunofluoroscence findings, and mesangial deposits, subendothelial deposits, and subepithelial deposits on electron microscopic findings. The children were treated with corticosteroids, ACE(angiotensin-converting enzyme) inhibitors, dipyridamole and other immunosuppressive agents. During the course of treatment, there were no children whose clinical condition worsened. Among 19 children, 3 children went into remission(2 in the nephrotic proteinuria group, 1 in the non-nephrotic proteinuria group) and 9 children went into a partial remission(4 in the nephrotic proteinuria group, 5 in the non-nephrotic proteinuria group) on urinalysis. There was no significant difference in the treatment results between the two groups. Conclusion : The 73.7% of children who were incidentally diagnosed with MPGN by the school urine screening program had reduced C3. 42.1% of the children had nephrotic range proteinuria. There were no significant differences in clinical features, laboratory test results, light microscopic, immunofluorescence microscopic, and electron microscopic findings between the nephrotic proteinuria group and the non-nephrotic proteinuria group except for the 24 hour urine protein secretion. Therefore, for early detection of MPGN during the school urine screening program, we strongly recommend a kidney biopsy if children have abnormal urine findings such as persistent proteinuria and persistent hematuria, or if the serum C3 is reduced.

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