• IMMUNE NETWORK
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• v.2 no.3
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• pp.166-174
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• 2002

Background: Human umbilical cord bloods, which could be taken during the delivery are utilized as a source of hematopoietic stem cells. Also in cord blood, there are several kinds of stem cells such as endothelial and mesenchymal stem cells. Methods: We isolated the mesenchymal stem cells from human umbilical cord bloods and confirmed the differentiation of these cells into osteoblast progenitor cells. The mesenchymal stem cells derived from umbilical cord blood have the ability to differentiate into specific tissue cells, which is one of characteristics of stem cells. These cells were originated from the multipolar shaped cells out of adherent cells of the umbilical cord blood mononuclear cell culture. Results: The mesenchymal stem cells expressed cell surface antigen CD13, CD90, CD102, CD105, ${\alpha}$-smooth muscle actin and cytoplasmic antigen vimentine. Having cultrued these cells in bone formation media, we observed the formation of extracellular matrix and the expression of alkaline phosphatase and of mRNA of cbfa-1, ostoecalcin and type I collagen. Conclusion: From these results we concluded that the cells isolated from the umbilical cord blood were mesenchymal stem cells, which we could differentiate into osteoblast when cultured in bone formation media. In short, it is suggested that these cells could be used as a new source of stem cells, which has the probability to alternate the embryonic stem cells.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.20 no.3
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• pp.250-255
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• 1998

The purpose of this study was to evaluate the histologic change of the inferior alveolar nerve according to distraction amount following mandibular lengthening. Seven rabbits weighing about 2 kg were used. Corticotomy was performed on the mandibular body anterior to the right first premolar region and unilateral external fixation device was placed. Every effort was made to preserve the inferior alveolar nerve during the corticotomy. The rabbits were then allowed to heal for 7 days without distraction of the device. The mandible was lengthened 0.36 mm/day, 0.76 mm/day, or 1.0 mm/day. Corticotomy and lengthening of mandible were not performed in control group. After the completion of the lengthening process, a 14-day-consolidation period was allowed. After consolidation, rabbits were sacrificed, and histologic examination of the inferior alveolar nerve was performed. The results obtained were as follows : 1. In the control group, normal trifascicular pattern of inferior alveolar nerve was observed. Epineurium, perineurium, endoneurium, and axon with myelin sheath were observed in normal appearance. 2. In 0.36 mm/day distraction group, the trifascicular pattern was normally shown, and there was no destruction in epineurium, perineurium, and endoneurium. The mild changes including myelin attenuation, axoplasmic swelling and darkening were observed. 3. In 0.72 mm/day distraction group, it was possible to differentiate the epineurium from the perineurium. Two normal fascicles and one injuried fascicle were observed with a partially destructed perineurium. Most of the axons had axoplasmic swelling and darkening. 4. In 1 mm/day distraction group, it was difficult to differentiate the nerve structures such as fascicles, epineurium, perineurium, and endoneurium. The axons were severely destroyed, except few which showed decreases in size and changes in shape. Some collagen matrices were observed around the axons. These results suggest that the higher the distraction amount, the more severe the injury to the inferior alveolar nerve, fascicles, axons. Although distraction osteogenesis may be useful, the amount of distraction should be carefully selected.

• The korean journal of orthodontics
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• v.42 no.6
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• pp.307-317
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• 2012

Objective: The purpose of this study was to investigate the isolation and characterization of multipotent human periodontal ligament (PDL) stem cells and to assess their ability to differentiate into bone, cartilage, and adipose tissue. Methods: PDL stem cells were isolated from 7 extracted human premolar teeth. Human PDL cells were expanded in culture, stained using anti-CD29, -CD34, -CD44, and -STRO-1 antibodies, and sorted by fluorescent activated cell sorting (FACS). Gingival fibroblasts (GFs) served as a positive control. PDL stem cells and GFs were cultured using standard conditions conducive for osteogenic, chondrogenic, or adipogenic differentiation. Results: An average of $152.8{\pm}27.6$ colony-forming units was present at day 7 in cultures of PDL stem cells. At day 4, PDL stem cells exhibited a significant increase in proliferation (p < 0.05), reaching nearly double the proliferation rate of GFs. About $5.6{\pm}4.5%$ of cells in human PDL tissues were strongly STRO-1-positive. In osteogenic cultures, calcium nodules were observed by day 21 in PDL stem cells, which showed more intense calcium staining than GF cultures. In adipogenic cultures, both cell populations showed positive Oil Red O staining by day 21. Additionally, in chondrogenic cultures, PDL stem cells expressed collagen type II by day 21. Conclusions: The PDL contains multipotent stem cells that have the potential to differentiate into osteoblasts, chondrocytes, and adipocytes. This adult PDL stem cell population can be utilized as potential sources of PDL in tissue engineering applications.

• Korean Journal of Plant Resources
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• v.14 no.2
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• pp.157-162
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• 2001

Korean native Orostachys species collected from 22 regions investigated morphological characters.O. japonicus species(No.3∼No.9) were similar leaf morphology and characterized by lanceolate leaves, cuspidate leaf apex.O. malacophyllus species(No.12∼No. 17) were morphologically characterized by obovate leaves and acute leaf apex. This species could be differentiate from other species by no thorn and leaf outlines formed by densely populated red dots.O. iwarenge species(No.18∼No.22) were diversified, such as obovate and elliptical leaves with acute, obtuse and round leaf apices. However, this species could be differentiate from other species bl'no thorn and grey powdery green colored leaves. Species collected from Maemuldo(No. 10) and Pocheon(No. 11) was assumed that these 2 species could be the new species which were not named classified.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.104-104
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• 2003

As an effort to direct differentiation of human embryonic stem (hES, MB03) cells to dopamine-producing neuronal cells, Nurr1 was transfected using conventional transfection protocol into MB03 and examined the expression of tyrosine hydroylase (TH) after differentiation induced by retinoic acid (RA) and ascorbic acid (AA). Experimentally, cells were transfected with linearized Nurr1 cDNA in pcDNA3.1 (+)-hygovernight followed by selection in medium containing hygromycin-B (150 $\mu$/ml). Expression of Nurr1 mRNA was confirmed by RT-PCR and protein by immunocytochemistry in the drug resistant clones. In order to study the effect of Nurr1 protein on the differentiation pattern of ES cells, one of the positive clones (MBNr24) was allowed to form embryoid body (EB) for 2 days and were induced to differentiate for another 4 days using RA (1 $\mu M$) and AA (50 mM) (2-/4+ protocol) followed by selection in N2 medium for 10 or 20 days. After 10 days in N2 medium, cells immunoreactive to anti-GFAP, anti-TH, or anti-NF200 antibodies were 38.8%, 11%, and 20.5%, respectively. After 20 days in N2 medium, cells expressing GFAP, TH, or NF200 were 28%, 15% and 44.8%, respectively but approximately 9% of MB03 expressed TH protein when the cells were induced to differentiate using a similar prorocol, These results suggest that ectopic expression of Nurr1 enhances generation of TH+ cells as well as neuronal cells when hES cells were differentiated by 2-/4+ protocol.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.60-60
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• 2003

Embryonic stem (ES) cells have property of self-renewal and can differentiate into the cells of all three primary germ layers. Recently, many growth factors, alteration of culture condition and gene modifications have been used to differentiate mouse and human ES cells into specific cell types. This study was performed to evaluate the differentiation protocol for human ES cells to the endodermal lineage cells. Human ES cells (Miz-hESl ) were cultured on STO feeder layer mitotically inactivated with mitemycin C, and embryoid bodies (EBs) were formed by suspension culture. Differentiation protocol of EBs consisted of three steps: stage I, culture of EBs for 6 days with ITSFn medium; stage II, culture of stage I cells for 8 days with N2 medium ; stage III, culture of stage II cells for 22 days with N2 medium. mRNA levels of the endodermal lineage differentiation genes were analyzed by semi- quantitative RT-PCR. The Oct-4 expression, a marker of the pluripotent state, was detected in undifferentiated human ES cells but progressively decreased after EBs formation. Differentiating human ES cells expressed marker genes of endodermal differentiation and pancreatic islet cells. GATA4, a-fetoprotein, Glut-2, and Ngn3 were expressed in all stages. However, albumin and insulin were expressed in only stage III cells. The human ES cells can be differentiated into endodermal lineage cells by multiple step culture system using various supplements. We are developing the more effective protocols for guided differentiation of human ES cells.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.97-97
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• 2003

Embryonic stem (ES) cells, derived from preimplantation embryos, are able to differentiate into various types of cells consisting the whole body, or pluripotency. In addition to the plasticity, ES cells are expected to be different from terminally differentiated cells in very many ways, such as patterns of gene expressions, ability and response of the cells in confronting environmental stimulations, metabolism, and growth rate. As a model system to differentiate these two types of cells, human ES (hES, MB03) cells and terminally differentiated cells (HeLa), we examined the ability of these two types of cells in confronting a severe oxidative insult, that is $H_2 O_2$. Ratio of dying cells as determined by the relative amount of dye neutral red entrapped within the cells after the exposures. Cell death rates were not significantly different when either MB03 or HeLa were exposed up to 0.4 mM $H_2 O_2$. However, relative amount of dye entrapped within the cells sharply decreased down to 0.12% in HeLa cells when the cells were exposed to 0.8 mM $H_2 O_2$, while it was approximately 54% in MB03. Pretreatment of cells with BSO (GSH chelator) and measurement of GSH content results suggest that cellular GSH is the major defensive mechanism of hES cells. Induction of apoptosis in hES cell was confirmed by DNA laddering, induction of Bax, and chromatin condensation. In summary, hES cells 1) are extremely resistant to oxidative stress, 2) utilize GSH as a major defensive mechanism. and 3) experience apoptosis upon exposure to oxidative stress.

• IMMUNE NETWORK
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• v.14 no.1
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• pp.54-65
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• 2014

Bone marrow-derived mesenchymal stem cells (MSCs) are multipotent, with the ability to differentiate into different cell types. Additionally, the immunomodulatory activity of MSCs can downregulate inflammatory responses. The use of MSCs to repair injured tissues and treat inflammation, including in neuroimmune diseases, has been extensively explored. Although MSCs have emerged as a promising resource for the treatment of neuroimmune diseases, attempts to define the molecular properties of MSCs have been limited by the heterogeneity of MSC populations. We recently developed a new method, the subfractionation culturing method, to isolate homogeneous human clonal MSCs (hcMSCs). The hcMSCs were able to differentiate into fat, cartilage, bone, neuroglia, and liver cell types. In this study, to better understand the properties of neurally differentiated MSCs, gene expression in highly homogeneous hcMSCs was analyzed. Neural differentiation of hcMSCs was induced for 14 days. Thereafter, RNA and genomic DNA was isolated and subjected to microarray analysis and DNA methylation array analysis, respectively. We correlated the transcriptome of hcMSCs during neural differentiation with the DNA methylation status. Here, we describe and discuss the gene expression profile of neurally differentiated hcMSCs. These findings will expand our understanding of the molecular properties of MSCs and contribute to the development of cell therapy for neuroimmune diseases.

• Phonetics and Speech Sciences
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• v.2 no.3
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• pp.157-164
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• 2010

This study investigated the characteristics of two aerodynamic indices, PTP (Phonation threshold pressure) and PTF (Phonation threshold flow) in patients with ADSD (adductor spasmodic dysphonia) and to see if two new aerodynamic indices can differentiate between normal and ADSD group. Additionally, PTP and PTF values were compared in terms of overall severity of ADSD in the patient group. The severity of ADSD was rated on a 7-point rating scale by two experienced speech language pathologists. The Kay Elemetrics Phonatory Aerodynamic System (PAS) (Kay Elemetrics Corp., Lincoln Park, NJ) was used to collect PTP and PTF measurements from 16 female normal subjects, 31 female patients with ADSD. Significantly lower PTF values (P< 0.05) were observed in ADSD when compared to those of normal control. Also, significantly lower PTF values in severe ADSD patients (P<.001). However, PTP could not distinguish patients with ADSD from control groups (P=0.119) and among the ADSD groups according to the severity (P=0.177). Consequently, PTF was more sensitive than PTP which might differentiate between normal speakers and ADSD and among different levels of severity within ADSD, suggesting that PTF could be a useful diagnostic parameter to measure the aerodynamic function of ADSD and provide the neurolaryngeal dysfunction in patients with ADSD.

• Clinical and Experimental Pediatrics
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• v.61 no.1
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• pp.12-16
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• 2018

Purpose: To differentiate adenoviral pharyngoconjunctival fever (PCF) from acute Kawasaki disease (KD) using laboratory tests before results of virus-real time polymerase chain reaction and ophthalmologic examination are obtained. Methods: Baseline patient characteristics and laboratory measurements were compared between 40 patients with adenovirus infection and 123 patients with KD. Results: The patients with adenovirus infection were generally older than those with KD (median: 3.9 years vs. 2 years, P=0.000). White blood cell and, platelet count, and aspartate aminotransferase, alanine aminotransferase, and N-terminal pro-brain natriuretic peptide (NT-proBNP) levels showed significant differences between the 2 groups, but the C-reactive protein (CRP) levels did not ($6.8{\pm}3.0mg/dL$ vs. $8.3{\pm}5.8mg/dL$, P=0.126). In the adenovirus infection group, the CRP levels were <1, <3, <10, and ${\geq}10mg/dL$ in 2 (5%), 3 (7.5%), 30 (75%), and 5 patients (12.5%), respectively. The cutoff NT-proBNP level was 265 pg/mL. Discrepancy was defined as CRP and NT-proBNP levels of ${\geq}3$ or <3 mg/dL, and <265 or ${\geq}265pg/mL$, respectively. Among the 35 patients with adenovirus infection whose CRP levels were ${\geq}3mg/dL$, 29 (82.9%) showed a discrepancy. Conversely, of the 103 patients with KD whose CRP levels were ${\geq}3mg/dL$, 83 (80.6%) showed no discrepancy. Between the groups, a significant difference in discrepancy rate was observed (P=0.000). None of the patients with adenovirus infection had CRP and NT-proBNP levels of <3 mg/dL and ${\geq}265pg/mL$, respectively. Conclusion: With a sensitivity of 82.9% and a specificity of 80.6%, CRP and NT-proBNP levels may differentiate between adenoviral PCF and acute KD.