• Clinical and Experimental Reproductive Medicine
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• v.26 no.1
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• pp.83-87
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• 1999

Since the blastocyst is broken and spreads out on a flat plastic culture dish (two dimensional culture) during in vitro development, it has been difficult to study the implantation process. It also has been difficult to analyse the interactions between endometrial epithelial and stromal cells because of the lack of a long-term in vitro model which can stimulate in vivo characteristics, as these cells eventually fail to proliferate or cease to express differentiated functions. Recently nontransformed cell lines, CUE-P and CUS-V2, derived from rat endometrial epithelium and stroma were reported. In this study, morphology of CUE-P and CUS-V2 was examined and oxytocin gene expression by CUE-P cells was demonstrated by RT-PCR. The CUE-P cells have a cuboidal morphology and CUS-V2 cells resemble fibroblast and exhibit a spindle-like morphology. In RT-PCR, same size of PCR products of oxytocin gene at hypothalamus, uterus and CUE-P cells were demonstrated. These results showed three dimensional culture system could be made by using the new cell lines.

• IMMUNE NETWORK
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• v.7 no.3
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• pp.117-123
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• 2007

Background: This study was designed to investigate the role of the hepatocyte growth factor (HGF) with regards to differentiation of somatic stem cells originating from the human umbilical cord blood (UCB) into hepatic lineage cells in vitro culture system. Methods: Mononuclear cells from UCB were cultured with and without HGF based on the fibroblast growth factor (FGF)-1, FGF-2, and stem cell factor. The cultured cells were confirmed by immunofluorescent staining analysis with albumin (ALB), cytokeratin-19 (CK-19), and proliferating cell nuclear antigen (PCNA) MoAb. ALB and CK-18 mRNA were also evaluated by reverse transcription-polymerase chain reaction. In order to observe changes in proliferating capacity with respect to the cultured period, CFSE with affinity to proliferating cells were tagged and later underwent flow cytometry. Results: In the HGF-treated group, cultured cells had a large oval shaped appearance with adherent, but easily detachable characteristics. In the HGF-non treated group, these cells were spindle-shaped with strong adherent characteristics. Expressions of ALB and CK-19 were evident in HGF-treated group compared to non-expression of those in to HGF-non treated group. Dual immunostaining analysis of the ALB producing cells showed presence of PCNA in their nuclei, and ALB and CK-18 mRNA were detected on the 21st day of cultured cells in the HGF-treated group. Conclusion: Our findings suggest that HGF has a pivotal role in differentiating somatic stem cells of human UCB into hepatic lineage cells in vitro.

• Polymer(Korea)
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• v.32 no.5
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• pp.409-414
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• 2008

The fabrication of gelatin nanofibers by electro spinning has been examined using the TFE/DW co-solvent system. It has been found that no beads-on-string structure was formed for the solution containing ionic salts. The resulting fibers exhibited a uniform diameter ranging from 110 to 125 nm. As the concentration of ionic salts increases, the beads become smaller and more spindle like, due to the increase of viscosity and conductivity. The addition of ionic salts induces a higher charge density on the surface of ejected jet during spinning, leading that higher elongation forces are applied to the jet. The higher enhancement of viscosity and conductivity was observed in gelatin solutions by the use of divalent salt. However, the concentration of ionic salts scarcely affected the variation of fiber diameter. While very low crystallinity was observed from XRD pattern for the sample containing no ionic salt, which increased with increasing the concentration of ionic salts.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.3
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• pp.175-184
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• 2022

Objective: The aim of this study was to evaluate the impacts of platelet-rich plasma (PRP) and conditioned medium (CM) derived from endometrial stromal cells on mouse preantral follicle culture in a two-dimensional system to produce competent mature oocytes for fertilization. Methods: In total, 240 preantral follicles were isolated from female mouse ovarian tissue and divided into four groups. The preantral follicles were isolated three times for each group and then cultured, respectively, in the presence of alpha minimum essential medium (control), PRP, CM, and PRP+CM. The in vitro growth, in vitro maturation, and cleavage percentage of the preantral follicles were investigated. Immunocytochemistry (IHC) was also conducted to monitor the meiotic progression of the oocytes. Additionally, the mRNA expression levels of the two folliculogenesis-related genes (Gdf9 and Bmp15) and two apoptosis-related genes (Bcl2 and Bax) were investigated using real-time polymerase chain reaction. Results: In the PRP, CM, and PRP+CM groups, the preantral follicle maturation (evaluated by identifying polar bodies) were greater than the control group. The cleavage rate in the CM, and PRP+CM groups were also greater than the control group. IHC analysis demonstrated that in each treatment group, meiotic spindle was normal. In the PRP+CM group, the gene expression levels of Bmp15, Gdf9, and Bcl2 were greater than in the other groups. The Bax gene was more strongly expressed in the PRP and control groups than in the other groups. Conclusion: Overall, the present study suggests that the combination of CM and PRP can effectively increase the growth and cleavage rate of mouse preantral follicles in vitro.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.3
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• pp.177-184
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• 2023

Objective: Reconstructed oocytes after polar body genome transfer constitute a potential therapeutic option for patients with a history of embryo fragmentation and advanced maternal age. However, the rescue of genetic material from the first polar body (PB1) through introduction into the donor cytoplasm is not yet ready for clinical application. Methods: Eighty-five oocytes were obtained following in vitro maturation (IVM) and divided into two groups: PB1 nuclear transfer (PB1NT; n=54) and control (n=31). Following enucleation and PB1 genomic transfer, PB1 fusion was assessed. Subsequently, all fused oocytes underwent intracytoplasmic sperm injection (ICSI) and were cultured in an incubator under a time-lapse monitoring system to evaluate fertilization, embryonic morphokinetic parameters, and cleavage patterns. Results: Following enucleation and fusion, 77.14% of oocytes survived, and 92.59% of polar bodies (PBs) fused. However, the normal fertilization rate was lower in the PB1NT group than in the control group (56.41% vs. 92%, p=0.002). No significant differences were observed in embryo kinetics between the groups, but a significant difference was detected in embryo developmental arrest after the four-cell stage, along with abnormal cleavage division in the PB1NT group. This was followed by significant between-group differences in the implantation potential rate and euploidy status. Most embryos in the PB1NT group had at least one abnormal cleavage division (93.3%, p=0.001). Conclusion: Fresh PB1NT oocytes successfully produced normal zygotes following PB fusion and ICSI in IVM oocytes. However, this was accompanied by low efficiency in developing into cleavage embryos, along with an increase in abnormal cleavage patterns.

• Journal of Veterinary Science
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• v.24 no.1
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• pp.4.1-4.16
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• 2023

Background: In vitro culture of preantral follicles is a promising technology for fertility preservation. Objectives: This study aims to investigate an optimized three-dimensional (3D) fetal bovine serum (FBS)-free preantral follicle culture system having a simple and easy operation. Methods: The isolated follicles from mouse ovaries were randomly divided in an ultra-low attachment 96-well plates supplement with FBS or bovine serum albumin (BSA) culture or encapsulated with an alginate supplement with FBS or BSA culture. Meanwhile, estradiol (E₂) concentration was assessed through enzyme-linked immunosorbent assay of culture supernatants. The diameter of follicular growth was measured, and the lumen of the follicle was photographed. Spindle microtubules of oocytes were detected via immunofluorescence. The ability of oocytes to fertilize was assessed using in vitro fertilization. Results: The diameters were larger for the growing secondary follicles cultured in ultra-low attachment 96-well plates than in the alginate gel on days 6, 8, and 10 (p < 0.05). Meanwhile, the E2 concentration in the BSA-supplemented medium was significantly higher in the alginate gel than in the other three groups on days 6 and 8 (p < 0.05), and the oocytes in the FBS-free system could complete meiosis and fertilization in vitro. Conclusions: The present study furnishes insights into the mature oocytes obtained from the 3D culture of the preantral follicle by using ultra-low attachment 96-well plate with an FBS-free system in vitro and supports the clinical practices to achieve competent, mature oocytes for in vitro fertilization.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.16 no.3
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• pp.321-347
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• 1994

Muscle spindle afferents from masseter muscle were labelled by the intra-axonal HRP injection and were processed for light microscopic reconstruction. Regions containing terminal arbors scattered in the central portion of the masseteric motor neuron pool (type I a) and those restricted to 2-3 small portion of it (type II) were selected and processed for electronmicroscopic analysis with serial sections. The shape of the labelled boutons was dome or elongated shape. Scalloped or glomerulus shape with peripherial indentation containing pre or postsynaptic neuronal propiles, which is occasionally found in the trigeminal main sensory nucleus and spinal dorsal horn, was not observed. Both type Ia and type II boutons had pale axoplasm and contained clear, spherical vesicles of uniform size(dia : 49-52nm) and occasionally large dense cored vesicles(dia : 87-118nm). The synaptic vesicles were evenly distributed throughout the boutons although there was a slight tendency of vesicles to accumulate at the presynaptic site. The average of short and long diameter(short D. + long D./2) of type I a bouton was smaller than that of type II bouton. All the labelled boutons, which showed prominent postsynaptic density, large synaptic area and multiple synaptic contact, made asymmetrical synaptic contact with postsynaptic neuronal propiles. Most of the type Ia and type II boutons made synaptic contact with only one neuronal propile and boutons which shows synaptic contact or more neuronal propiles was not observed. Most of the type Ia boutons(87.2%) were presynaptic to the soma or proximal dendrite and a few remainder(12.8%) made synaptic contact with dendritic shaft or distal dendrite. In contrast, majority of type II boutons showed synaptic contact with dendritic shaft and remainder with soma or proximal dendrite. In conclusion, terminal boutons which participate in the excitatory monosynaptic jaw jerk reflex made synaptic contact with more proximal region of the neuron, and showed very simple synaptic connection, compared with those from the primary afferenst in the other region of the central nervous system such as spinal dorsal horn and trigeminal main sensory nucleus which assumed to be responsible for the mediating pain, tactile sensation, sensory processing or sensory discrimination.

• Journal of the Korea Concrete Institute
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• v.16 no.2 s.80
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• pp.241-248
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• 2004

The rheological properties of cement paste system mixed with mineral admixtures (MAs) used to increase the strength and improve durability and fluidity of concrete were investigated. And cement paste systems were designed as one-, two- and three-ingredients blended paste systems. The rheological properties of paste systems were assessed by Rotovisco RT 20 rheometer (Hakke inc.) having a cylindrical serrated spindle. The rheological properties of one-ingredient paste systems were improved with increasing the dosage of superplasticizer. For two-ingredients paste systems, as increasing the replacement ratio of blast furnace slag (BFS) and fly ash (FA), the yield stress and plastic viscosity were decreased compared with non-replacement. In the ordinary portland cement (OPC)-silica fume (SF) paste systems, in accordance with an increase in the replacement ratio of SF, the yield stress and plastic viscosity were increased steeply. For three-ingredients paste systems, both OPC-BFS-SF and OPC-FA-SF paste systems, the rheological properties were improved compared with the only replacement of SF. In the case of both two-and three-ingredients paste systems, the rheological properties using BFS were improved more than FA.

• Korean Journal Plant Pathology
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• v.1 no.3
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• pp.199-206
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• 1985

Viroids are the smallest. well-characterized infectious agents presently known. and so far viroids have been found only in higher plants. The structures of viroid-molecules are single-stranded, covalently closed circular RNA molecules with a range of 240 to 380 nucleotides according to the various viroids. Viroids are remarkable not only as a new category of pathogen, which cause economically important diseases, but also as an excellent model system for biochemical and biophysical investigations because of their small size, relative stability and their self-replication. Four different patato spindle tuber viroid isolates, which express the different symptoms on the same host plant exchange only 2 to 6 nucleotides in the total number of 359 nucleotides, but now the mechanism of viroid pathogenicity is not explained fully. Viroid-melecules are replicated without any special viroid-associated proteins, and during the process of viroid replication oligomeric viroid-associated RNAs are detected at nuclei of viroid infected leaf tissue. The mechanism of viroid replication can now be illustrated according to a possible explanation of rolling-circle system. Although the rapid progress have been made in elucidation of the biochemical and biophysical properties of PSTV and other viroids, the mechanism of viroid replication and pathogenicity is less known and is still a matter of speculation. When these problems can be sufficiently explained, the viroid molecule could play an important role as an available vector in plant genetic engineering.

• Korean Journal of Heritage: History & Science
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• v.38
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• pp.285-303
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• 2005

Through the excavated excellent articles of the Tang era, we have considered the building techniques and styles of a square pavilion, an octagonal pavilion, a miniature hill, a pond, etc., could read building techniques of the scenic structures on ancient literatures including records and additionally about their poetical life at that time and inquired into the arrangement, structure, building techniques, etc. of a miniature hill, a pond, a pavilion, a flowerplant, etc., on Nakyangmyungwonki. Thus, under the research on the private landscape architecture, especially the nobility gardening, around excavated excellent articles and literatures, 'A miniature hill(假山) and a pond for landscaping views formed the center of a garden, and additionally a pavilion was built and flowerplants were set. The miniature hill of laying stones and having a carven, steeping, stratifying, looking like dyed green and birds' singing among hills and eating water on the lakeshore${\ldots}$' was expressed. The pond of digging in the ground and conducting water had its water system developed. There were several kinds of pavilions such as 廳, 堂, 館, 亭, 臺, 樓, 閣, 榭, etc. As examples of landscaping plants, there were a bamboo, a lotus flower, a peony, aromatic trees a pine, a korean spindle tree, a big cone pine, an empress tree, a wild walnut, a peach, a plum, a Japanese apricot tree, an apricot tree, a chrysanthemum, arrowroot vines, etc. Thus, the garden of the Tang era, abundant, diverse and excellent, enjoyed the prime of the period of prosperity. Due to cultural exchange, it is supposed that the period of united Shilla of the same age would meet with the period of prosperity in the developmental history of Korea landscape, based on the nobility garden system '4 different dwelling-houses every season on a record "四節游宅"'.