• Clinical and Experimental Pediatrics
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• v.52 no.9
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• pp.1015-1020
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• 2009

Purpose:The aim of this study is to explore the effect of the Toll-like receptor 9 (TLR9) expressed in plasmacytoid dendritic cells (pDCs) that respond to antigen to Th2 immune deviation in allergic patients. Methods:Subjects consisted of 19 allergic patients and 17 healthy volunteers. Skin prick tests and nasal provocation tests were performed for the two groups. Peripheral blood mononuclear cells (PBMCs) were collected from subjects and analyzed for the Lineage Cocktail (CD3, CD14, CD16, CD19, CD20, CD56) (-), HLA-DR (+), and CD123 (+) using flow cytometry. In addition, we analyzed TLR9 mRNA by reverse transcriptase-polymerase chain reaction. The level of $interferon-{\alpha}$ ($IFN-{\alpha}$) of the PBMCs following stimulation with the TLR9 ligand CpG-ODN 2216 was also evaluated. Results:Analyses of CD123 (+) revealed a nearly similar distribution for the classical pDC markers in the allergic group ($0.1%{\pm}0.04%$) and in the controls ($0.25%{\pm}0.23%$). The mRNA levels of TLR9 on PBMCs were not different between the allergic group and the controls ($1.29{\pm}0.41$ vs. $1.25{\pm}0.23$, respectively). Additionally, the level of $IFN-{\alpha}$ in PBMCs exposed to stimuli of the TLR9 ligand CpG-ODN 2216 was not significantly different between the two groups ($911{\pm}829$ vs. $1,095{\pm}888pg/mL$, respectively). Conclusion:We found no evidence that TLR9-dependent immune responses in human pDCs are associated with allergic status.

• The Journal of Korean Physical Therapy
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• v.22 no.3
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• pp.79-85
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• 2010

Purpose: Many trials for new therapeutic approaches such as stem cell-based transplantation have been conducted to improve the repair and regeneration of injured cord tissue and to restore functions following spinal cord injury (SCI) in animals and humans. Adipose tissue-derived stromal cells (ATSCs) have multi-lineage potential to differentiate into cells with neuron-like morphology. Most studies of stem cell transplantation therapy after SCI are focused on cellular regeneration and restoration of motor function, but not on unwanted effects after transplantation such as neuropathic pain. This study was focused on whether transplantation of ATSCs could facilitate or attenuate hindpaw pain responses to heat, cold and mechanical stimulation, as well as on improvement of locomotor function in a rat with SCI. Methods: A spinal cord injury rat model was produced using an NYU impactor by dropping a 10 g rod from a height of 25 mm on to the T9 segment. Human ATSCs (hATSCs; approximately $5{\times}10^5$ cells) or DMEM were injected into the perilesional area 9 days after the SCI. After transplantation, hindpaw withdrawal responses to heat, cold and mechanical allodynia were measured over 7 weeks. Motor recovery on the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale and on the inclined plane test were also evaluated. Results: The present study demonstrated that increased hindpaw withdrawal responses to cold allodynia was observed in both groups after transplantation, but the development of cold-induced allodynia in the hATSC transplantation group was significantly larger than in the control group. The difference between the two groups in locomotor functional improvement after SCI was also significant. Conclusion: Careful consideration not only of optimal functional benefits but also of unintended side effects such as neuropathic pain is necessary before stem cell transplantation therapy after SCI.

• Clinical and Experimental Reproductive Medicine
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• v.2 no.2
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• pp.13-26
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• 1975

Seven aspects of attitude toward marriage in Korea are examined to better understand present and future marriage patterns. Also, various facets of current marriage practice are compared with attitudes. The study comprises three groups of roughly 600 women each, selected by random sampling from a rural, an urban, and a semi-urban area. A carefully designed and pretested questionnaire was checked for reliability by a reinterview in a 15% subsample. The great majority of Korean women support traditional attitudes that one must or should marry. The small group who recommend that one should not marry are mostly the very young or the never married, whose attitudes still may change. However, there are important and probably predictive shifts in favor of more individual decision, especially among the better educated, the young, and the more urban. Traditional reasons for marriage such as "custom" and procreation are ranked first by a majority, but there is a large shift to more contemporary or liberal desire for companionship and love, also primarily among the better educated, the urban, the young, and the never married. The traditional attitude that parents should have the sole or major role in mate selection is still held by a bare majority; the educated, urban, young, and never married are more liberal. Only 6% opt for each of the two extremes: That the parent alone or the respondent alone should decide. The remainder prefer one of the two middle-of-the-road positions where parent and child together decide. The proportions of respondents who classed specified criteria as moat important for selecting a husband, arranging the criteria in order from traditional to contemporary were: Lineage, etc., 23%; personal attributes, 40%; health and education, 27%; and love, 10%. The changing attitudes are suggested by the fact that love was ranked first by only 3% of the poorly educated rural poulation versus 23% of urban college level and 31% of the urban never married. There has been a substantial rise in the ideal age of marriage over the past twelve or more years, but there also is evidence that the ideal age is at or near a ceiling. Knowledge about legal age of marriage is minimal; the implications of this for proposed legislation are discussed. Three-fifthes to four-fifths of all respondents married husbands of the same religious, residential, and economic backgrounds as themselves. Almost all of them married men of the same or higher educational level. These evidences of traditional influences in mate selection are contrasted with the low priority given some of those items in earlier questions on reasons for marriage and criterion for selecting husband. Contrary to the expressed attitudes as to who should select the husband, we find that marriages of the study sample were stated to be arranged by parents alone in 62%; and in another 23%, the parents made the decision but asked the respondent's views. Such arrangements were most frequent among the rural, the less educated, and the older respondents and less common in the urban and more educated. The implications of these and related findings are discussed.

• Reproductive and Developmental Biology
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• v.36 no.1
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• pp.33-37
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• 2012

Differential capacity of the parthenogenetic embryonic stem cells (PESCs) is still under controversy and the mechanisms of its neural induction are yet poorly understood. Here we demonstrated neural lineage induction of PESCs by addition of insulin-like growth factor-2 (Igf2), which is an important factor for embryo organ development and a paternally expressed imprinting gene. Murine PESCs were aggregated to embryoid bodies (EBs) by suspension culture under the leukemia inhibitory factor-free condition for 4 days. To test the effect of exogenous Igf2, 30 ng/ml of Igf2 was supplemented to EBs induction medium. Then neural induction was carried out with serum-free medium containing insulin, transferrin, selenium, and fibronectin complex (ITSFn) for 12 days. Normal murine embryonic stem cells derived from fertilized embryos (ESCs) were used as the control group. Neural potential of differentiated PESCs and ESCs were analyzed by immunofluorescent labeling and real-time PCR assay (Nestin, neural progenitor marker; Tuj1, neuronal cell marker; GFAP, glial cell marker). The differentiated cells from both ESC and PESC showed heterogeneous population of Nestin, Tuj1, and GFAP positive cells. In terms of the level of gene expression, PESC showed 4 times higher level of GFAP expression than ESCs. After exposure to Igf2, the expression level of GFAP decreased both in derivatives of PESCs and ESCs. Interestingly, the expression level of $Tuj1$ increased only in ESCs, not in PESCs. The results show that IGF2 is a positive effector for suppressing over-expressed glial differentiation during neural induction of PESCs and for promoting neuronal differentiation of ESCs, while exogenous Igf2 could not accelerate the neuronal differentiation of PESCs. Although exogenous Igf2 promotes neuronal differentiation of normal ESCs, expression of endogenous $Igf2$ may be critical for initiating neuronal differentiation of pluripotent stem cells. The findings may contribute to understanding of the relationship between imprinting mechanism and neural differentiation and its application to neural tissue repair in the future.

• Biomolecules & Therapeutics
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• v.20 no.2
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• pp.158-164
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• 2012

Inhibiting the bioactivities of circulating endothelial progenitor cells (EPCs) results in significant inhibition of neovessel formation during tumor angiogenesis. To investigate the potential effect of phloroglucinol as an EPC inhibitor, we performed several in vitro functional assays using $CD34^+$ cells isolated from human umbilical cord blood (HUCB). Although a high treatment dose of phloroglucinol did not show any cell toxicity, it specifically induced the cell death of EPCs under serum free conditions through apoptosis. In the EPC colony-forming assay (EPC-CFA), we observed a significant decreased in the small EPC-CFUs for the phloroglucinol group, implying that phloroglucinol inhibited the early stage of EPC commitment. In addition, in the in vitro expansion assay using $CD34^+$ cells, treatment with phloroglucinol was shown to inhibit endothelial lineage commitment, as demonstrated by the decrease in endothelial surface markers of EPCs including $CD34^+$, $CD34^+/CD133^+$, $CD34^+/CD31^+$ and $CD34^+/CXCR4^+$. This is the first report to demonstrate that phloroglucinol can inhibit the functional bioactivities of EPCs, indicating that phloroglucinol may be used as an EPC inhibitor in the development of biosafe anti-tumor drugs that target tumor angiogenesis.

• Journal of Life Science
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• v.26 no.7
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• pp.868-874
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• 2016

Receptor-interacting protein kinase 1 (RIPK1) and RIPK3 are members of the serine or threonine protein kinase superfamily that phosphorylates the hydroxyl group of serine or threonine through the highly conserved kinase region. The RIPK family plays a crucial role not only in inflammation and innate immunity, but also in mediating programmed cell death, such as apoptosis and necroptosis. The interaction between RIPK1 and other TNFR1-related proteins has been shown to assemble a signaling complex I that controls activation of the pro-survival transcription factor NF-κB upon binding of cytokines to TNF receptor 1 (TNFR1). Moreover, RIPK1 and RIPK3 interact through their RIP homotypic interaction motifs (RHIMs) to mediate programmed necrosis, which has long been considered an accidental and uncontrolled cell death form with morphological characteristics differing from those of apoptosis. Highly conserved sequences of RHIM in RIPK1 and RIPK3 were shown to regulate their binary interaction, leading to assembly of a cytosolic amyloid complex termed the “necrosome”. The necrosome also contains mixed lineage kinase domain-like protein (MLKL), which has been found recently to be a substrate of RIPK3 to mediate downstream signaling. This review provides an overview of the functional and physiological characteristics of RIPKs and MLKL in TNF signaling.

• Clinics in Shoulder and Elbow
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• v.15 no.2
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• pp.65-72
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• 2012

Purpose: To evaluate the differentiation potential of stem cells and their immunophenotype from 3 different sources. Methods: Our study involved three stem cell sources-subacromial bursal tissue, bone marrow, and umbilical cord blood. We obtained the subacromial bursal tissue and bone marrow from the patients undergoing shoulder surgery. After collecting the sample, we applied specific induction media for neurogenic, adipogenic and osteogenic differentiation. Also, flow-cytometry analysis was done to reveal the cell surface antigens. Results: We obtained 100% (8 cases) neural and adipogenic differentiation, but 62.5% (5 of 8 cases) osseous differentiation among the subacromial bursal tissue group. Bone marrow derived cells showed 100% neural (6 cases) and adipogenic (5 cases) differentiation, but 80% (4 of 5 cases) osseous differentiation. Umbilical cord blood derived cells revealed 97% (65 of 67 cases) neural, 53.7% (29 of 54 cases) adipogenic and 68.4% (39 of 57 cases) osseous differentiation. Immunophenotype analysis revealed that surface markers of bone marrow, subacromial bursal cell and umbilical cord blood derived mesenchymal stem cells are different from each other. Conclusions: Mesenchymal stem cells are potential agents in regenerative medicine and are characterized by expression of surface markers and by their differentiation potential. Our study with stem cells from subacromial bursal tissue, bone marrow and umbilical cord discovered that each stem cell has unique differentiation potential and function based on its origin. Various stem cells show multi-lineage differentiations in vitro which can be correlated to in vivo conditions.

• The korean journal of orthodontics
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• v.31 no.6 s.89
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• pp.589-600
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• 2001

Insulin-like growth factor I (IGF-I) has the local tissue regulating actions. In bone, IGF-I increases the replication of osteoblastic lineage, probably preosteoblasts, and enhances osteoblastic collagen synthesis and matrix composition rates. The purpose of this study was to investigate the local regulatory effect of IGF-I on periodontium totally, both in an autocrine and paracrine manner. To examine the effect of IGF-I directly on osteoblast (OB) of test rats, and indirectlv on OB via periodontal ligament fibroblast (PDLF), and the effect of gingival fibroblast (GF) on OB via cellular paracrine manner for the understanding of humoral action of adjacent tissue, GF and PDLF were obtained from male Sprague-Dawley rats of six to eight weeks of age. OB was obtained iron frontal and parietal calvarial bone of Sprague-Dawley 21day-old-fetus. After each tell was Incubated 24 hours, for collecting conditioned medium, different concentrations of IGF-I (1,10,100 ng/ml,1ml/well) was adding in the GF, PDLF cells, and the supernatant from these cultures was put into the primary OB culture with $1{\times}10^4$cell/ml/well. The experimental group was divided into six groups control OB, IGF-I treated OB, OB culture with conditioned medium from PDLF, OB culture with conditioned medium from IGF-I treated PDLF, OB culture with conditioned medium from GF, OB culture with conditioned medium from IGF-I treated GF. After final IGF-I treatment, OB was Incubated for 24 hours, and alkaline phosphatase activity assay, BMP expression, cell proliferation measurement using MTT assay, total protein measurement, Collagen synthesis assay using western blot, and examination of bone nodule synthesis were done. Alkaline phosphatase expressions were increased in the group of PDLF-IGF-I supernatant treatment. Direct IGF-I treatment with concentrations of 100ng/m1 showed increased viable tell number measured by MTT assay. And IGF-I treatment did not increase total protein amount. The entire experimental group showed BMP2, 4 expression in western blot, and there was no significant difference between control and experimental groups. These results suggested that supernatant from PDLF effects on increasing cellular activities of OB regardless of IGF-I, and at high concentration, IGF-I increases OB tell proliferation.

• Journal of the korean academy of Pediatric Dentistry
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• v.28 no.2
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• pp.238-246
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• 2001

Bone morphogenetic proteins(BMPs), members of the transforming growth factor $\beta$(TGF-$\beta$) superfamily were first identified as the factors that induce ectopic bone formation in vivo, when implanted into muscular tissue. Especially BMP-2 inhibits terminal differentiation of C2C12 myoblasts and converts them into osteoblast lineage cells. In the molecular mechanism of the signal transduction of TGF-$\beta$ and related factors, intracellular signaling proteins were identified as Smad. In previous study, it has been reported that Smad 1 and Smad 5, which belong to the R-Smad family mediate BMP signaling, were involved in the induction of osteoblast differentiation in C2C12 cells. To understnad the role of Smads involved in osteogenic transdifferentiation in C2C12 cell, in present study, after we stably transfected C2C12 cells with each. Smad(Smad 1,Smad 5) expression vector, cultured for 3 days and stained for alkaline phophatase activity. ALP activity positive cells appeared in the Smad 1, Smad 5 stably transfected cell even in the abscence of BMP. After transiently co-transfected C2C12 cells with each Smad expression vector and ALP promoter, it was examined that Smad 1 and Smad 5 expression vector had increased about 2 fold ALP promoter activity in the abscence of BMP. These result suggested that both Smad 1 and Smad 5 were involved in the intracellular BMP signals which induce osteoblast differentiation in C2C12 cells. The effect of BMP on C2C12 cells with Smad 1, Smad 5 transfected were studied by using northern blot analysis. the treatment of BMP upregulated ALP mRNA level in three groups, especially upregulation of ALP was larger in Smad 1, Smad 5 transfected cell than control group. Pretreatment with cycloheximide($10{\mu}g/ml$), a protein synthesis inhibitor resulted in blocking the ALP gene expression even in BMP(100ng/ml) treated cell. These results suggested that Smad increased the level of ALP mRNA via the synthesis of a certain transcriptional regulatory protein.

• Journal of the Korean Institute of Traditional Landscape Architecture
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• v.34 no.3
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• pp.78-97
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• 2016

The objectives of this study are to examine the context of the establishment of Suhoe Gugok, Byeokgye Gugok Vally, and Nosan Palkyung, which have been established in Seojong-myeon of Yangpyeong-gun, by literature review and site investigations, and to determine the sceneries of Byeokgye scenic site as enjoyed and managed during the period of Hwaseo Lee, Hang-ro(華西 李恒老). The results of the study are as follows. First, Byeokgye Gugok Vally(黃蘗九曲) and Nosan Palkyung(蘆山八景), which have been established after the period of Hwaseo and theorized to have been established around key scenic areas associated with Hwaseo's activities, the analysis results showed that they were collecting sceneries of modern times. The extensive overlap between Byeokgye Gugok Vally and concentrated scenic elements of Suhoe Gugok(水回九曲), and the artificial configuration from the end point of Suhoe Gugok to the beginning point of Nosan Palkyung, reveal the pattern of space conflict and hegemony between Byeokgyes of Suip-ri and Nomun-ri. This is likely to be caused by the conflict between the historicity of the group that enjoyed Byeokgye prior to Hwaso's period and the strong territoriality of the space filled with the image of Hwaseo. Second, Byeokgye Gugok Vally was the secondary spatial system created by selecting the most scenic sites in Suip-ri while expanding the area of Nosan Palkyung. After establishment of Byeokgye Gugok Vally, the spatial identity of the entire Byeokgyecheon area was effectively established. This was a "Hwaseo-oriented" move, including the complete exclusion of the scenic sites from the pre-Hwaseo period such as Cheongseo Gujang and Suhoe Gugok's Letters Carved on the Rock. Consequently, the entire Byeokgyecheon area was reorganized into a cultural scenic site with Heoseo's influence. Third, Fifth, creations of Gugok(九曲) to determine the lineage of the Hwaseo School from Juja(朱子) to Yulgok(栗谷) to Uam(尤庵) to Hwaseo is likely to be an opportunity of birth and external motivation of the establishment of new Gugok Palkyung. In other words, Nosan Palkyung and Byeokgye Gugok Vally are likely to have been created as a reaction to the change of the center of the Hwaseo School to Okgyedong, and with strategic orientation based on the motivation and needs such as creation of the connecting space between Mui Gugok, Gosan Gugok, and Okgye Gugok, and the elevation of Hwaseo's status. Fourth, from the Hwaseo's Li-centric point of view, all revered sites in Beokwon(蘗園) that he managed existed as the spatial creative work to experience the existence of "li" through the objects in the landscape and the boundary of the spirit of emptiness of the aesthetic self. This clearly shows how Byeokgye Gugok Vally or Nosan Palkyung must be defined, and furthermore, appreciated and approached, prior to discussing it as the space associated with Hwaseo. Fifth, Nosan Palkyung was composed of cultural scenic landscapes of Gokjungkyung(曲中景) with eight scenic sites where Hwaseo gave his teachings and spend time around, in the Byeokgye of Nomun-ri area of Byeokgye Gugok Vally. The sceneries is, however, collected by depending on Hwaseo's Letters Carved on the Rock and poetry. Consequently, an inner exuberance of Nosan Palkyung is satisfied beside Byeokgye Gugok Vally, but its conceptual adequacy leaves room for questions.