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Ex Vivo Expansion of Hematopoietic Stem/Progenitor Cells by Coculture using Insert

  • Kim, Kyung-Suk;Kim, Haekwon;Do, Byung-Rok;Park, Seah;Kwon, Hyuck-Chan;Kim, Hyun-Ok;Im, Jung-Ae
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.77-77
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    • 2003
  • Coculture of HSC with bone marrow-derived mesenchymal stem cells (BM-MSCs) is one of used methods to increase cell numbers before transplant to the patients. However, because of difficulties to purify HSCs after coculture with BM-MSCs, it needs to develop a method to overcome the problem. In the present study, we have examined whether a culture insert placed over a feeder layer might support the expansion of HSCs within the insert. $CD34^+/ $ cells isolated from the umbilical cord blood by using midiMACS were divided into three groups. A group of 1 $\times$ $10^5$ cells were grown on a culture insert without feeder layer (Direct). The same number of HSCs was directly cocultured with BM-MSCs (Contact). The third group was placed onto an insert below which BM-MSCs were grown (Insert). To distinguish feeder cells from HSCs, BM-MSCs was pre-labeled fluorescently with PKH26 and 1 $\times$ $10^5$ cells were seeded in the culture dishes. After culture for 13 days, the expansion factor (x) of HSCs that were grown without feeder layer (Direct) was $26.6 \pm 8.4.$ In contrast, the number of HSCs directly cocultured with feeder layer was 59.6 $\pm$ 0.5 and that of HSCs cultured onto an insert was $46.9 \pm 8.4.$ The percentage of BM-MSCs cells remained being fluorescent was $97.9 \pm 0.3%$ after culture. Immune-phenotypically large proportion of cultured cells were founded to be differentiated into myeloid/monocyte progenitor cells. The ability of BM-MSCs, fetal lung, cartilage and brain tissue cells to support ex vivo expansion of HSCs was also examined using the insert. After 11 days of coculture with each of these cells, the expansion factor of HSCs was 15.0, 39.0, 32.0 and 24.0, respectively. Based upon these observations, it is concluded that the coculture method using insert is very effective to support ex vivo expansion of HSCs and to eliminate the contamination of other cells used to coculture wth HSCs.

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Status and Changes in Chemical Properties of Paddy Soil in Gangwon Province

  • Yoon, Byeong-Sung;Choi, Seung-Chul;Lim, Soo-Jeoung;Heo, Su-Jeong;Kim, In-Jong;Kang, Seong-Soo
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.4
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    • pp.293-299
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    • 2016
  • The chemical properties of paddy fields in Gangwon province were monitored every 4 years from 2003 to 2015 in order to provide basic information for soil fertility management of paddy fields. In 2015, the soil chemical properties of paddy fields were 5.9 in pH, $22g\;kg^{-1}$ in organic matter (OM), and $123mg\;kg^{-1}$ in available (Avail.) phosphate $P_2O_5$. Exchangeable (Exch.) potassium (K), calcium (Ca) and magnesium (Mg) were 0.39, 4.8 and $0.9cmolc\;kg^{-1}$, respectively, and Avail. $SiO_2$ was $170mg\;kg^{-1}$. In the long-term analysis, the contents of Avail. $SiO_2$, Exch. K and pH of paddy soils showed increasing tendency. However, Avail. $P_2O_5$, Exch. Ca and Mg tended to decrease, and there were no significant changes in the contents of OM. Soil OM, Avail. $P_2O_5$ and $SiO_2$ were not different among the different topographical sampling sites. However, the mean value of Exch. K and Ca were different among the different topographical sampling sites, and exceeded optimal values in the fluvio-marine plains. Different soil texture resulted in different soil pH, while no difference for OM, Avail. $P_2O_5$ and $SiO_2$. Paddy soil samples within appropriate pH range increased from 65% in 2003 to 77% in 2007, 68% in 2011, and 71% in 2015. In case of Avail. $SiO_2$, soil samples within appropriate range increased from 20% in 2003, to 37% in 2007, 29% in 2011, and 45% in 2015. Meanwhile, Cd and Pb were distributed to less than 5% of soil pollution standards. Cu, As and Zn were distributed to less than 10%, 15% and 20%, respectively. Therefore, paddy soil in Gangwon Province was judged to be safe. As a result, paddy fields with more or less in nutrient level need to be fertilized based on the soil analysis. And the application of silicate fertilizer is strongly recommended to those of paddy fields in need. In addition, soil management including the cultivation of green manure crop or application of rice straw is necessary to increase the organic matter content of paddy soil.

The Characteristic of Strength for a Lime Stone in Donghae Area and Harden Cement Milk of Super Injection Grouting (동해 석회암과 SIG 고결체의 강도특성)

  • Park, Young-Ho;Kim, Nak-Young;Hong, Sa-Myun;Yook, Jeong-Hoon;Kim, Ki-Seog
    • Proceedings of the Korean Geotechical Society Conference
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    • 2004.03b
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    • pp.137-145
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    • 2004
  • Limestone zone in korea have been distributed to diagonal line so that it is wide from the Gangwondo to the Jeonlanamdo. The limestone cavity and fractured zone were formed by chemical weathering. Limestone cavity and fractured zone was reinforced with cemented milk(w/c=60%)by high pressure jet grouting by tripple -pipe to establish bridge foundation on the ground condition like limestone cavity. To analyze property of limestone and solid of cement milk(w/c=65%), mixed solid of cement, core NX size in the limestone cavity and fractured zone and compressive strength. Seismic tomograpy exploration was pcrforn1cd to analyze deformation modulus of limestone. The analysis suggests that deformation modulus of limestone has effect on uniaxial compressive strength, seismic velocity, seismic elasticity modulus. Average static elasticity modulus of limestone is $5.08{\times}10^5kgf/cm^2$, cement and coal mixed solid is $0.25{\times}10^5kgf/cm^2$, $0.095{\times}10^5kgf/cm^2$. Average seismic velocity of limestone is 5.240m/sec, cement and coal mixed solid is 2,211.3m/sec, 1,447.5m/sec. Average uniaxial compressive strength of limestone was $1,221.3kgf/cm^2$ and limestone specimen mixed with cement milk and solid of cement milk mixed with coal were $125.22kgf/cm^2$, $35kgf/cm^2$ each other. Average friction angle of limestone was $49.14^{\circ}$ and limestone specimen mixed with cement milk and solid of cement milk mixed with coal were $38.39^{\circ}, 25.83^{\circ}$ each other. Average cohesion of limestone was $137.7kgf/cm^2$ and limestone specimen mixed with cement milk and solid of cement milk mixed with coal were $23.5kgf/cm^2$, $15.5kgf/cm^2$ each other. Average deformation modulus of limestone was $2.84{\times}10^5kgf/cm^2$ and limestone specimen mixed with cement milk and solid of cement milk mixed with coal were $0.4{\times}10^5kgf/cm^2, 0.12{\times}10^5kgf/cm^2$ each other. It was analyzed that the elasticity and uniaxial compressive strength, seismic velocity of solid of cement milk mixed limestone pieces and coal had an highly interrelation regardless of existence of limestones pieces and coal but it had shown that limestones had an lower interrelation. In case of field seismic velocity and deformation of limestone, SIC solid of cement milk mixed with coal and limestone pieces had an highly interrelation.

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The Prognostic Significance of Tumor Angiogenesis and Expression of Vascular Endothelial Growth Factor in Papillary Carcinomas of the Thyroid Gland (갑상선 유두상 암종에서 종양혈관형성 및 혈관내피성장인자 발현의 예후인자적 의의)

  • Kang Hun-Dae;Kim Seong-Bae;Kim Tae-Hyun;Oh Sang-Hoon;Yoon Hye-Kyong;Kim Sang-Hyo
    • Korean Journal of Head & Neck Oncology
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    • v.20 no.2
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    • pp.135-142
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    • 2004
  • Objectives: The purpose of this study was to evaluate for prognostic significance of VEGF expression and tumor angiogenesis in papillary carcinomas of the thyroid. Materials and Methods: The materials were 79 cases of papillary thyroid carcinomas, and age, sex, tumor size, multiplicity of tumor, capsular invasion, lymph node metastasis, recurrence, TNM stage, DeGroot stage and AMES scale were evaluated. An immunohistochemical stains for CD 34 to estimate microvessel density (MVD), and VEGF were done. MVD was defined as an average count of vessels per ${\times}400$ power field in the most vascularized area. VEGF expression was interpreted as 1+ and 2+ according to staining intensity and percentages of positive cells. Results: Mean score of MVD was $39.7{\pm}16.9.$ MVD were significantly higher in cases with capsular invasion (p=0.0001), lymph node metastasis (p=0.0001), TNM stage III (p=0.0022), DeGroot stage III (p=0.0163) and high risk group by AMES scale (p=0.0001). VEGF 2+ expression rate was significantly increased in cases with capsular invasion and lymph node metastasis (p=0.0006, p=0.0013), and in cases with TNM stage III, DeGroot stage III and high risk group by AMES scale (p=0.0236, p=0.0003, p=0.0293). In VEGF 2 + expression group, MVD was significantly higher than in VEGF 1 + group (p=0.0008), and MVD showed positive relation to VEGF 2 + expression (r=0.4616). Conclusion: VEGF expression and high MVD were significantly correlated to capsular invasion, lymph node metastasis, TNM stage III, DeGroot stage III and high risk group by AMES scale. The expression of VEGF and high MVD could be considered to be one of prognostic factor in papillary thyroid carcinomas.

A Study on the Removal of Heavy Metals from Groundwater Using Permeable Reactive Barriers Based on Nano FeS (나노 FeS를 이용한 투과성반응벽체의 중금속 오염 지하수 처리에 관한 연구)

  • Jung, Gwan-Ju;Choi, Sang-Il;Lee, Jai-Young
    • Journal of Soil and Groundwater Environment
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    • v.14 no.6
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    • pp.19-28
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    • 2009
  • The acid mine drainage (AMD) and landfill leachates released into the subsurface environment can result in serious environmental problems like soil and groundwater contamination. The AMD and the leachates of landfill were known to contain many heavy metals. In this study, the author assessed the reactivity and ability of the FeS coated-ALC for the removal of contaminants (As, Cd, Cu, Pb, Ni, Zn, Al) in AMD and leachates in landfill. The synthetic nano-FeS and Autoclaved Lightweight Concrete (ALC) were used as reactive materials in the permeable reactive barriers(PRBs). The result of batch test indicated that synthetic nano-FeS can remove 99% of heavy metals for the 1hr of reaction time except for As and Ni(about 90%). However, the 80% of As and Ni was removed in column 1(FeS coated-ALC). The column 2(Ore FeS) removed more than 99% of heavy metals. The pH of the column 1 was increased from 3.51 to 6.39~6.50, and the pH with column 2 was increased from 3.51 to 9.20. As the result of this study, the author can surmise that the synthetic nano-FeS coated ALC will use as a very good reactive material of the PRBs to treat the contaminated groundwater with AMD and leachate of landfill.

Medico-Legal Consideration of Hemopneumothorax - Closing Claim Study- (${\cdot}$기흉과 관련된 의료법학적 문제에 대한 고찰 -종결된 사건을 중심으로-)

  • Bae, Hyu-Na;Cheon, Young-Jin
    • Journal of Chest Surgery
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    • v.39 no.2 s.259
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    • pp.117-126
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    • 2006
  • Background: The purpose of this study is to describe the characteristics of malpractice claims related to hemopneumothorax and to identify the causes and potential preventability of such claims. Material and Method: A retrospective study was performed by reviewing the records in the Lawnb website and Lx CD-rom: the records on closed malpractice claims involving hemopneumothorax were abstracted from the files available for analysis. The records were reviewed and were analysed to determine the etiology of hemopneumothorax, patient age, results of lawsuit and indemnity payment, underlying diseases, cause of death or complications, and the factors associated with a successful defense. Result: Seven closed claim involving hemopneumothorax were founded in the data for malpractice. Three claims were supreme court decision, one was a high court decision and three claims were district court decision. The most common cause of death was tension pneumothorax. Four of which resulted in indemnity payments. Conclusion: While malpractice claims involving hemopneumothorax were uncommon, they resulted in a high rate and amount of indemnity payments. Claims are more common in pediatric patients. In case of iatrogenic hemopneumothorax, post-procedural X-ray can improve patient outcome and is also associated with decreased indemnity risks. Informed consent is also important.

Dependence of $O_2$ Plasma Treatment of ITO Electrode on Electrical and Optical Properties of Polymer Light Emitting Diodes (ITO 투명전극의 $O_2$ 플라즈마 처리가 고분자 유기발광다이오드의 전기.광학적 특성에 미치는 영향)

  • Gong, Su-Cheol;Back, In-Jea;Yoo, Jea-Huyk;Lim, Hun-Sung;Yang, Sin-Huyk;Shin, Sang-Bea;Shin, Ik-Seup;Chang, Gee-Keun;Chang, Ho-Jung
    • Journal of the Korean institute of surface engineering
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    • v.39 no.3
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    • pp.93-97
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    • 2006
  • Polymer light emitting diodes (PLEDs) are expected to be commercialized as next generation displays by advantages of the fast response time, low driving voltage and easy manufacturing process for large sized flexible display. Generally, the electrical and optical properties of PLEDs are affected by the surface conditions of transparent electrode. The PLED devices with ITO/PEDOT:PSS/PVK/PFO-poss/LiF/Al structures were prepared by using the spin coating method. For this, PEDOT:PSS(poly(3,4-ethylenedioxythiophene):poly(styrene sulfolnate)) Al 4083 and PVK(N-vinylcabozole) were used as hole injection and transport layers. The PFO-poss(poly(9,9-dioctylfluorene)) was used as the emitting layer. The dependence of $O_2$ plasma treatment of ITO electrode on the electrical and optical properties of PLEDs were investigated. The sheet resistances increased slightly with an improved surface roughness of ITO electrode as the RF power increased during $O_2$ plasma treatment. The PLED devices prepared on the ITO/Glass substrates, which were plasma-treated at 40 watt in RF power for 30 seconds under 40 mtorr $O_2$ pressure, showed the maximum external emission efficiency of 0.86 lm/W and the maximum luminance of $250\;cd/m^2$, respectively. The CIE color coordinates are ranged $X\;=\;0.13{\sim}0.18$ and $Y\;=\;0.10{\sim}0.16$, showing blue color. emission.

Isolation of New CHO Cell Mutants Defective in CMP-Sialic Acid Biosynthesis and Transport

  • Shin, Dong-Jun;Kang, Ji Young;Kim, Youn Uck;Yoon, Joong Sik;Choy, Hyon E;Maeda, Yusuke;Kinoshita, Taroh;Hong, Yeongjin
    • Molecules and Cells
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    • v.22 no.3
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    • pp.343-352
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    • 2006
  • Sialic acid is a sugar typically found at the N-glycan termini of glycoproteins in mammalian cells. Lec3 CHO cell mutants are deficient in epimerase activity, due to a defect in the gene that encodes a bifunctional UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE). Sialic acid modification on the cell surface is partially affected in these cells. We have mutagenized Lec3 CHO cells and isolated six mutants (termed C2m) deficient in the cell surface expression of polysialic acid (PSA). Mutant C2m9 was partially defective in expression of cell-surface PSA and wheat germ agglutinin (WGA) binding, while in the other five mutants, both cell-surface PSA and WGA binding were undetectable. PSA expression was restored by complementation with the gene encoding the CMP-sialic acid transporter (CST), indicating that CST mutations were responsible for the phenotypes of the C2m cells. We characterized the CST mutations in these cells by Northern blotting and RT-PCR. C2m9 and C2m45 carried missense mutations resulting in glycine to glutamate substitutions at amino acids 217 (G217E) and 256 (G256E), respectively. C2m13, C2m39 and C2m31 had nonsense mutations that resulted in decreased CST mRNA stability, and C2m34 carried a putative splice site mutation. PSA and CD15s expression in CST-deficient Lec2 cells were partially rescued by G217E CST, but not by G256E CST, although both proteins were expressed at similar levels, and localized to the Golgi. These results indicate that the novel missense mutations isolated in this study affect CST activity.

Mucosal Immunity Related to FOXP3+ Regulatory T Cells, Th17 Cells and Cytokines in Pediatric Inflammatory Bowel Disease

  • Cho, Jinhee;Kim, Sorina;Yang, Da Hee;Lee, Juyeon;Park, Kyeong Won;Go, Junyong;Hyun, Chang-Lim;Jee, Youngheun;Kang, Ki Soo
    • Journal of Korean Medical Science
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    • v.33 no.52
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    • pp.336.1-336.12
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    • 2018
  • Background: We aimed to investigate mucosal immunity related to forkhead box P3 ($FOXP3^+$) regulatory T (Treg) cells, T helper 17 (Th17) cells and cytokines in pediatric inflammatory bowel disease (IBD). Methods: Mucosal tissues from terminal ileum and colon and serum samples were collected from twelve children with IBD and seven control children. Immunohistochemical staining was done using anti-human FOXP3 and anti-$ROR{\gamma}t$ antibodies. Serum levels of cytokines were analyzed using a multiplex assay covering interleukin $(IL)-1{\beta}$, IL-4, IL-6, IL-10, IL-17A/F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, interferon $(IFN)-{\gamma}$, soluble CD40L, and tumor necrosis factor-${\alpha}$. Results: $FOXP3^+$ Treg cells in the lamina propria (LP) of terminal ileum of patients with Crohn's disease were significantly (P < 0.05) higher than those in the healthy controls. $ROR{\gamma}t^+$ T cells of terminal ileum tended to be higher in Crohn's disease than those in the control. In the multiplex assay, serum concentrations (pg/mL) of IL-4 ($9.6{\pm}1.5$ vs. $12.7{\pm}3.0$), IL-21 ($14.9{\pm}1.5$ vs. $26.4{\pm}9.1$), IL-33 ($14.3{\pm}0.9$ vs. $19.1{\pm}5.3$), and $IFN-{\gamma}$ ($15.2{\pm}5.9$ vs. $50.2{\pm}42.4$) were significantly lower in Crohn's disease than those in the control group. However, serum concentration of IL-6 ($119.1{\pm}79.6$ vs. $52.9{\pm}39.1$) was higher in Crohn's disease than that in the control. Serum concentrations of IL-17A ($64.2{\pm}17.2$ vs. $28.3{\pm}10.0$) and IL-22 ($37.5{\pm}8.8$ vs. $27.2{\pm}3.7$) were significantly higher in ulcerative colitis than those in Crohn's disease. Conclusion: Mucosal immunity analysis showed increased $FOXP3^+$ T reg cells in the LP with Crohn's disease while Th17 cell polarizing and signature cytokines were decreased in the serum samples of Crohn's disease but increased in ulcerative colitis.

Changes of Kidney Injury Molecule-1 Expression and Renal Allograft Function in Protocol and for Cause Renal Allograft Biopsy (이식신 계획생검 및 재생검에서 Kidney Injury Molecule-1 표현과 이식신 기능 변화)

  • Kim, Yonhee;Lee, A-Lan;Kim, Myoung Soo;Joo, Dong Jin;Kim, Beom Seok;Huh, Kyu Ha;Kim, Soon Il;Kim, Yu Seun;Jeong, Hyeon Joo
    • Korean Journal of Transplantation
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    • v.28 no.3
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    • pp.135-143
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    • 2014
  • Background: Kidney injury molecule-1 (KIM-1) is known as a good ancillary marker of acute kidney injury (AKI) and its expression has also been observed in acute rejection and chronic graft dysfunction. We tested usefulness of KIM-1 as an indicator of acute and chronic renal graft injury by correlating KIM-1 expression with renal graft function and histology. Methods: A total of 133 zero-time biopsies and 42 follow-up biopsies obtained within 1 year posttransplantation were selected. Renal tubular KIM-1 staining was graded semiquantitatively from 0 to 3 and the extent of staining was expressed as the ratio of KIM-1 positive/CD10 positive proximal tubules using Image J program. Results: KIM-1 was positive in 39.8% of zero-time biopsies. KIM-1 positive cases were predominantly male and had received grafts from donors with older age, deceased donors, and poor renal function at the time of donation, compared with KIM-1 negative cases. KIM-1 expression showed correlation with delayed graft function and acute tubular necrosis. In comparison of KIM-1 expression between stable grafts (n=23) and grafts with dysfunction (n=19) at the time of repeated biopsy, the intensity/extent of KIM-1 staining and renal histology at zero-time did not differ significantly between the two groups. Histologically, KIM-1 expression was significantly increased with both acute and chronic changes of glomeruli, tubules and interstitium, peritubular capillaritis, and arteriolar hyalinosis. Conclusions: KIM-1 can be used as an ancillary marker of AKI and a nonspecific indicator of acute inflammation and tubulointerstitial fibrosis. However, KIM-1 expression at zero-time is not suitable for prediction of long-term graft dysfunction.