• Korean Journal of Clinical Laboratory Science
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• v.51 no.2
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• pp.125-133
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• 2019

A microscope is the fundamental research and diagnostic apparatus for clinical investigation of signaling transduction, morphological changes and physiological tracking of cells and intact tissues from patients in the biomedical laboratory science. Proper use, care and maintenance of microscope with comprehensive understanding in mechanism are fully requested for reliable image data and accurate interpretation for diagnosis in the clinical laboratory. The standard operating procedure (SOP) for light microscopes includes performance procedure, brief information of all mechanical parts of microscopes with systematic troubleshooting mechanism depending on the laboratory capacity. Maintenance program encompasses cleaning objective, ocular lenses and inner optics; replacement and calibration of light source; XY sample stage management; point spread function (PSF) measurement for confocal laser scanning microscope (CLSM); quality control (QC) program in fluorescent microscopy; and systematic troubleshooting. Laser safety is one of the concern for medical technologists engaged in CLSM laboratory. Laser safety guideline based on the laser classification and risk level, and advisory lab wear for CLSM users are also expatiated in this overview. Since acquired image data presents a wide range of information at the moment of acquisition, well-maintained microscopes with proper microscopic maintenance program are impulsive for its interpretation and diagnosis in the clinical laboratory.

• Nuclear Medicine and Molecular Imaging
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• v.41 no.3
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• pp.226-233
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• 2007

Purpose: Dual reporter gene imaging has several advantages for more sophisticated molecular imaging studies such as gene therapy monitoring. Herein, we have constructed hepatoma cell line expressing dual reporter genes of sodium iodide symporter (NIS) and enhanced green fluorescence protein (EGFP), and the functionalities of the genes were evaluated in vivo by nuclear and optical imaging. Materials and Methods: A pRetro-PN vector was constructed after separating NIS gene from pcDNA-NIS. RSV-EGFP-WPRE fragment separated from pLNRGW was cloned into pRetro-PN vector. The final vector expressing dual reporter genes was named pRetro-PNRGW. A human hepatoma (HepG2) cells were transfected by the retrovirus containing NIS and EGFP gene (HepG2-NE). Expression of NIS gene was confirmed by RT-PCR, radioiodine uptake and efflux studies. Expression of EGFP was confirmed by RT-PCR and fluorescence microscope. The HepG2 and HepG2-NE cells were implanted in shoulder and hindlimb of nude mice, then fluorescence image, gamma camera image and I-124 microPET image were undertaken. Results: The HepG2-NE cell was successfully constructed. RT-PCR showed NIS and EGFP mRNA expression. About 50% of cells showed fluorescence. The iodine uptake of NIS-expressed cells was about 9 times higher than control. In efflux study, $T_{1/2}$ of HepG2-NE cells was 9 min. HepG2-NE xenograft showed high signal-to-background fluorescent spots and higher iodine-uptake compared to those of HepG2 xenograft. Conclusion: A hepatoma cell line expressing NIS and EGFP dual reporter genes was successfully constructed and could be used as a potential either by therapeutic gene or imaging reporter gene.

• Journal of Biomedical Engineering Research
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• v.41 no.2
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• pp.67-74
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• 2020

Brain tumors or gliomas are fatal cancer species with high recurrence rates due to their strong invasiveness. Therefore, the goal of surgery is complete tumor resection. However, the surgery is difficult to distinguish the border because tumors and blood vessels have the same color tone and shape. The fluorescein sodium is used as a fluorescence contrast agent for boundary separation. When the external light source is irradiated, yellow fluorescence is expressed in the tumor, which helps distinguish between blood vessels and tumor boundaries. But, the fluorescence expression of fluorescence sodium depends on the concentration of fluorescein sodium and such analytical data is insufficient. The unclear fluorescence can obscure the boundaries between blood vessels and tumors. In addition, reduce the efficiency of fluorescence sodium use. This paper proposes a protocol of concentration range for fluorescence expression conditions. Fluorescent expression was observed using a near-infrared (NIR) color camera with corresponding dilution using normal saline in 1 ml microtube. The flunoresence emission density range is 1.00 mM to 0.15 mM. The fluorescence emission begin to 1.00 mM and the 0.15 mM discolor. The discolor is difficult to fluorescence emission condition obserbation. Thus, the maximum density range of the bright fluoresecein is 0.15 mM to 0.30 mM. When the concentration range of fluorescein sodium is analyzed based on the gradient of fluorescence expression and the power measurement, the brightest fluorescence is expected to facilitate the complete resection of the tumor. For the concentration range protocol, setting concentration ranges and analyzing fluorescence expression image according to saturation and brightness to find optimal fluorescence concentration are important. Concentration range protocols for fluorescence expression conditions can be used to find optimal concentrations of substances whose expression pattern varies with concentration ranges. This study is expected to be helpful in the boundary classification and resection of brain tumors and glioma.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.6
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• pp.511-520
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• 2010

Purpose: This study was conducted in order to compare the efficacy of osseointegration of three different calcium metaphosphate (CMP) coated implants in the rabbit's femur. Materials and Methods: Twenty four rabbits and three different type of CMP coated implants and RBM implants were used in this study. The animals were divided into 4 groups on the basis of implant surface characteristics. Two implants were installed into the condyle of femur of each rabbits. The animals were sacrificed at 2 and 4 weeks after installation. The undecalcified specimens were prepared for histological, radiological examination and histomorphometric analysis of implant-bone contact ratios (BIC) and bone area ratio (BA). Results: Two implants were failed to osseointegrate and implant success rate was 95.2%. There were not any significant inflammatory response in all groups. Fluorescent image at 4 weeks shows that remodeling is slower in RBM group than CMP group. CMP III showed more active remodeling than CMP I, II. In histomorphologic analysis, BIC ratio at 2 weeks was lower than 4 weeks. Conclusion: The results suggest that the ratios of CMP coated implants were higher than that of RBM control group but there is no significantly difference between RBM group and CMP group. In conclusion, CMP coated implant had more clinical availability than RBM implants.

• Archives of Plastic Surgery
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• v.34 no.6
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• pp.679-684
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• 2007

Purpose: DMH(1,2-dimethylhydrazine) has been known to induce vascular neoplasm such as malignant endothelioma in animal experiment, through induction of abnormal proliferation of HUVECs. In our previous studies, 11 types of PKC isoenzymes were determined by RT-PCR and the expression of $PKC{\alpha}$, and ${\mu}$ was more prominent than other PKC isoenzymes in the DMH-treated group. However, this result was not based on objective assessment. In this study, we further evaluated the role of $PKC{\alpha}$ on the DMH-induced abnormal proliferation of HUVECs by two different methods to identify its presence with high relevance in objective view. $PKC{\mu}$ will be investigated in further study. Methods: The study was conducted with the cultured HUVECs group(control) and the $0.75{\times}10^{-9}M$ DMH-treated group. After processing protein extraction in 0 and 24 hour, extracted protein was treated of quantitative test through BCA protein assay. In the western blot analysis, electrophoresis was performed in the order of gel preparation, sample preparation, and gel running. Electrotransfer to nitrocellulose membrane and reaction with antibody were done. Detection of $PKC{\alpha}$ was achieved through "Gel Image Analysis System". In the fluorescence immunocytochemical analysis, the grading of radiance of the intracellular $PKC{\alpha}$ particles was detected with confocal microscope after treating with primary and fluorescent secondary antibody in 0 and 24 hours. Results: The Western blot analysis showed increased $PKC{\alpha}$ expression from the specimen obtained in 24 hour of the DMH treatment group when compared to those in control group. Under confocal fluorescence microscope, the emitting radiance in the DMH treated group was brighter at 24 hours as well. Conclusion: We believe that $PKC{\alpha}$ plays a role in DMH-induced abnormal proliferation of the vascular endothelium, which may provide insights in understanding the vascular neoplasm.

• The Journal of Korean Academy of Prosthodontics
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• v.39 no.6
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• pp.659-681
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• 2001

Platelet-rich plasma(PRP) has been known to increase the rate and degree of bone formation by virtue of growth factors in concentrated platelets. Although its great healing effect on bone defect or pre-implantation site preparation in conjunction with bone substitute has been reported, the effect associated with implant is unknown. The purpose of this study was to investigate the effect of PRP on rapid osseointegration of endosseous dental implants in the rabbit tibiae. Twenty two adult female New Zealand white rabbits, weighing approximately 2.7-3.3kg, were used for this study. Twelve of the 22 animals were used for histomorphometric analysis and ten of the 22 were for removal torque test. Each animal received two implants in each tibia (two treated with PRP and two as control) and was given fluorochrome intramuscularly. For histomorphometric analysis, rabbits were divided into four groups according to the healing period. At 1 week, 2 weeks, 4 weeks and 8 weeks postoperatively, each three animals were sacrificed serially and the amount and rate of bone formation around dental implant were examined on the undecalcified sections under fluorescent microscope, polarized microscope and light microscope connected to a personal computer equipped with image analysis system. For removal torque test, rabbits were divided into two groups and removal torque tests were performed at 4 weeks, 10 weeks after implant placement. In total, 88 screw shaped, commercially pure titanium implants (Neoplant, Neobiotech, Seoul, Korea) were used in this study. Labeling pattern reflected differences of two groups in bone formation rate at each period. Histomorphometrically, PRP group showed significantly higher bone volume within threads compared to control group at 2 weeks ($70.30{\pm}4.96%$ vs. $50.68{\pm}6.33%$; P < .01) and 4 weeks ($82.59{\pm}5.94%$ vs. $72.94{\pm}4.57%$; P < .05 ). PRP group at 1, 2 and 4 weeks revealed similar degree of bone volume formation comparable to control group at 2, 4 and 8 weeks, respectively. On the other hand, while PRP group showed higher bone-implant contact ($47.37{\pm}8.09%$) than control group ($33.16{\pm}13.47%$) at 2 weeks, there were no significant differences between PRP group and control group for any experimental period. Removal torque values also showed no significant differences between PRP group and control group at any experimental period (P > .05). These findings imply that PRP could induce rapid, more bone formation around implant during early healing period and get faster secondary stability for reducing healing period, though it has not induced bone maturation enough to resist functional loading.

• Journal of Korean Biological Nursing Science
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• v.1 no.1
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• pp.25-41
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• 1999

Physiological activity of osteoblast including bone formation is known to be closely related to the increase of intracellular $Ca^{2+}$ activity($[Ca^{2+}]_i$) in osteoblast. $Ca^{2+}$ is an important intracellular messenger in diverse cellular functions, and regulation of its level is mediated by the transmembrane $Ca^{2+}$ movement via $Ca^{2+}$ channels, $Na^+-Ca^{2+}$ exchange, and by intracellular $Ca^{2+}$ movement through the intracellular stores. The purpose of this study is to investigate how the intracellular $Ca^{2+}$ is regulated in osteoblast-like cells(OLCs) by measuring $Ca^{2+}$ activity with cell imaging technique. OLCs were isolated from femur and tibia of neonatal rats, and cultured for 7 days. Cultured OLCs were loaded with a $Ca^{2+}$-sensitive fluorescent dye, Fura-2, and fluorescence images were monitored with a cooled CCD camera. The images were processed and analyzed with an image analyzing software. The results were as follows. (1) $[Ca^{2+}]_i$ of OLC decreased as the $Ca^{2+}$ concentration in the superfusing Tyrode solution was lowered. When $Na^+$ concentration in the superfusing solution was decreased, $[Ca^{2+}]_i$ increased.. These suggest that $Ca^{2+}$ flux occurs via the $Na^+-Ca^{2+}$ exchange mechanism. (2) When $Na^+$ in the superfusing solution was removed. a transient $Ca^{2+}$, increase($Ca^{2+}$ spike) was occasionally observed. However, $Ca^{2+}$ spike was not observed after adding 1 ${\mu}M$ thapsigargin. This implies that the generation of $Ca^{2+}$ spike is mediated by the release of $Ca^{2+}$ from endoplasmic reticulum(ER). (3) As the $Ca^{2+}$ concentration in the superfusing solution was raised, the frequency of 0mM $Na^+$-induced $Ca^{2+}$ spike increased, suggesting that $Ca^{2+}$-induced $Ca^{2+}$ release(CICR) mechanism exists. (4) After $[Ca^{2+}]_i$ was decreased with the superfusion of $Ca^{2+}$-free solution containing thapsigargin, the recovery of $[Ca^{2+}]_i$ with reperfusion of 2.5mM $Ca^{2+}$ solution transiently exceeded the control level, suggesting that the depletion of $Ca^{2+}$ in ER induces $Ca^{2+}$ influx from extracellular medium via store-operated $Ca^{2+}$ influx(SOCI) mechanism. (5) $[Ca^{2+}]_i$ was not affected by the superfusion of 25mM $K^+$ Tyrode solution. These results suggest that intracellular $Ca^{2+}$ activity in osteoblast is regulated by transmembrane $Ca^{2+}$ flux via $Na^+-Ca^{2+}$ exchange, $Ca^{2+}$ release from the internal store (ER) via $Ca^{2+}$-induced $Ca^{2+}$ release, and store-operated $Ca^{2+}$ influx across the cell membrane.

• Science of Emotion and Sensibility
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• v.17 no.2
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• pp.35-44
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• 2014

The purpose of this paper is to investigate the readability of Nearly White background color in display environment. To perform the experiment, warmup experiment was carried out to for cognitive of whiteness using a Tablet PC. The experiment environment was by a indoor window seat under a mixture of fluorescent light and sunlight. The result of previous test was that the most whiteness perception was a color white a medium wavelength and followed by the short wavelength. The main experiment was to find readability, color combination and visual strains using 4 nearly whites background colors (Reddish, Greenish, Blueish, Yellowish) and one white background color, also text color is limited on black. As a result, readability and color combination had the highest score against a Greenish white background. Visual strain is also the lowest against a Greenish background. On the contrary, readability and color combination preference has the lowest score against the Reddish background. Visual strain is also the highest against a Reddish background. We propose the background color, which improves readability and reduces visual strain in visual display terminals environment through the research results.

• Journal of Veterinary Clinics
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• v.27 no.3
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• pp.216-219
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• 2010

The purpose of this study is to establish experimental canine skin barrier disruption model, the study was designed to observe calcium ion in skin frozen tissue of canine skin and also the modulation of calcium ion distribution of normal skin with disrupted skin such as clipping, acetone, tape stripping damages according to time. To compare the changes of calcium ion gradient after damages, the distribution of calcium ion in the canine epidermis was visualized by blotting to gel containing chemical indicator (Calcium Green-1) with fluorescent microscope and the effects of skin barrier damages were examined according to time. Three mins and 1hr after acetone damage, the gradations of epidermis and hair follicle showed more radiant and disappeared after 48 hrs. On the contrary, 3mins and 1hr after tape stripping damage, the gradations showed more radiant than those of acetone damage, and these gradations were stabilized after 48 hrs. The method we presented here could show the visual image of the calcium ions in frozen tissue without further preparation, and it might be useful to investigate the role of calcium ion in the canine epidermal barrier recovery, however, it might be need further methodological improvement to get accurate quantitative information.

• Restorative Dentistry and Endodontics
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• v.25 no.1
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• pp.1-16
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• 2000

It is difficult to treat the endodontic apical perforation successfully. In this study, we hypothesized that the application of PDGF-BB and IGF-I into periapical perforation site may accelerate periapical healing and lead to bone deposition. And the specificity of osteonectin in periapical healing was investigated. The experiments were performed on the upper and lower 51 premolar teeth of 4 beagle dogs. The pulp chamber of each tooth was opened and the dental plaque was inserted into the canal for developing the periapical lesion for 5 weeks. Then, the roots were artificially perforated at the apex with the number 4 profile of .06 taper. In each step, standard periapical radiographs were taken to compare the size of lesion each other. The radiographs were scanned and analyzed by image analysis system. The mean and standard deviation of periradicular radiolucency ratios were calculated in each group. ANOVA was used for comparison. 51 premolars were grouped into 3 groups; control group, calcium hydroxide-treated group and calcium hydroxide plus growth factors-treated group. In the control group, the apical perforations were not sealed and obturated with gutta-percha and ZOE sealer by lateral condensation technique. In the experimental groups, the apical perforation were sealed with calcium hydroxide and with/without $4{\mu}g$ of PDGF-BB & IGF-I in cellulose gel and obturated by lateral condensation technique. Fluorescent bone markers were used to measure new bone formation. Following 2, 4, 12 weeks after experiment the dogs were sacrificed and histologic sections were prepared. Each tooth block including periapical lesion was sectioned mesiodistally. One half of the sections were decalcified with 6% nitric acid and processed by standard paraffin embedding technique. The sections were stained by hematoxylin and eosin, and immunostained for osteonectin. Histomorphometrical measurement of neoformed bone was performed using a light microscope. And the other half of the sections were prepared by undecalcified preparation, and confocal laser scanning microscopic investigations were done.