• 한국가시화정보학회:학술대회논문집
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• 2004.12a
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• pp.119-124
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• 2004

Experimental study was conducted to characterize the flow effect of particle migration in a microchannel which can be used to deliver small amount of liquids, drugs, biological agents and particles in microfluidic devices. Fluorescent particles of $1\{mu}m$ diameter were used to obtain velocity profiles of the fluid in which large particles of $10\{mu}m$ diameter were suspended at different volume fraction of 0.6 and $0.8\%$. Measurements were obtained by using micro-PIV system which contains a Nd:YAG laser with a light of 532-nm wavelength, an inverted epi-fluorescent microscope and a cooled CCD camera to record particle images. The volume fraction of $\phi$ and the particle Reynolds number $Re_p$Rep were used as a parameter to assess the influence of the velocity profile of the suspensions. To expect the slip velocity between the particle and fluids, experiments were carried out at low volume fraction. It was shown that the velocity profile was not influenced by Rep but influenced by the volume fraction, which is in similar trend with the previous study.

• Restorative Dentistry and Endodontics
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• v.25 no.4
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• pp.606-618
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• 2000

Dental pulp infection is most commonly caused by extensive dental caries, and some bacterial species invade root canals; bacterial components and products are thought to be associated with the pathogenesis of periapical periodontitis. A principle driving force behind pulpal disease response appears to lie in the host immune system's to bacteria and their products. We examined the production of interleukin $1{\beta}$ (IL-$1{\beta}$) and tumor necrosis factor ${\alpha}$(TNF-${\alpha}$) from human peripheral mononuclear cells, lymphocytes and monocytes stimulated by heat-killed Acitnobacillus actinomycetemcomitans (ATCC 29523), Porphyromonas gingivalis (ATCC 33277) and Prevotella intermedia (ATCC 25611), and also by their sonicated bacterial extracts (SBE), respectively. The effects of three strains of heat-killed bacteria and their SBEs on the morphology of cultured blood cell lines HL-60 (KCLB 10240) and J774A.1 (KCLB 40067) were observed under the inverted microscope. Ultrastructural changes of J774A.1 exposed to heat-killed P. intermedia and its SBE were investigated using transmission electron microscopy. Production of IL-$1{\beta}$ was reduced in human peripheral mononuclear cells after stimulation by sonic bacterial extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia. Heat-killed and sonic extract of P. gingivalis inhibited the production of TNF-${\alpha}$ in peripheral mononuclear cells. Production of TNF-${\alpha}$ was inhibited in peripheral monocytes after stimulation by sonic extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia. HL-60 and J 774A.1 cells showed granular degeneration after treatment with heat-killed and sonic extracts of A. actinomycetemcomitans, P. gingivalis, and P. intermedia Chromatin margination and shrinkage were observed in 774A.1 treated with heat-killed P. intermedia. Cell wall structure and organelles were destroyed and vacuoles were formed in cytoplasm in J774A.1 treated with P. intermedia sonic extract. These results suggest that A actinomycetemcomitans, P gingivalis and P intermedia may have an important role in the formation and progression of pulpal diseases via both modulation of production of IL-$1{\beta}$ and TNF-${\alpha}$ from blood mononuclear cells and cytopathic effects.

• Restorative Dentistry and Endodontics
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• v.29 no.6
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• pp.498-503
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• 2004

The properties of ideal root canal sealers include the ability of sealing the total root canal system and no toxic effects to periradicular tissues. Cytotoxicity test using cell culture is a common screening method for evaluation of the biocompatibility of root canal sealers. The purpose of this study was to investigate the cytotoxic effect of newly developed resin-based sealer (Adseal 1, 2, and 3) comparing with those commercial resin-based sealers (AH26 and AH Plus), ZOE-based sealers (Tubliseal EWT, Pulp Canal Sealer EWT) and calcium hydroxide based sealer (Sealapex), An indirect contact test of cytotoxicity by agar diffusion was performed according to the international standard ISO 10993-5. L929 fibroblast cells were incubated at $37^{\circ}C$ in humidified 5% $CO_2-containing$ air atmosphere. The freshly mixed test materials were inserted into glass rings of internal diameter 5 mm and height 5 mm placed on the agar. After the 24 hrs incubation period, the decolorization zones around the test materials were assessed using an inverted microscope with a calibrated screen. A Decolorization Index was determined for each specimen. Adseal 1. 2, and 3 did not exert any cytotoxic effects, whereas AH26, AH Plus, Tubliseal EWT, Pulp Canal Sealer EWT, and Sealapex produced mild cytotoxicity.

• Journal of Veterinary Clinics
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• v.22 no.4
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• pp.322-327
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• 2005

DLA class I complement-dependent cytotoxicity (CDC) cross-match method was established to control hyperacute rejections in organ transplantation. The aim of the present study is to investigate if DLA class 1 CDC corssmatch method is effective to prevent hyperacute rejections in canine renal allografts. Seven mongrel dogs of similar age and weight were used. Erythrocyte crossmatch was first performed and only the negatives were used. Among the same blood types, CDC cross-match was performed. Anti-dog serum, Hank's balanced salt solution(HBSS), and tile self-serum was used as a positive-, a negative-, and all auto-control respectively. After the reaction with class I complement, it was stained with eosin and scored by international cytotoxicity scoring system under inverted phase microscope. According to these results, kidneys oi CDC negatives among same blood types were cross-transplanted to observe the incidence of hyperacute rejections. One of four 1.2 B blood type dogs had autoantibodies. here were negative CDC results among each blood type, and also there were negative results between different blood types. Two pairs with the same blood types and negative CDC results underwent allo-transplantation each other. There were no hyperacute rejections. DLA cross-match method studied in this experiment for canine renal allograft can be effective to prevent hyperacute rejections. it may be applicable for the future studies of histocompatibility testing in canine renal allografts.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.2
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• pp.139-147
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• 2002

Objective : This study was to establish a reproducible differentiation system from the parthenogenetic mouse embryonic stem (P-mES02) cells into functional cardiomyocytes like as in vitro fertilization mouse embryonic stem (mES01) cells. Materials and Methods: To induce differentiation, P-mES02 cells were dissociated and aggregated in suspension culture environment for embryoid body (EB) formation. For differentiation into cardiomyocytes, day 4 EBs were treated with 0.75% dimethyl sulfoxide (DMSO) for another 4 days (4-/4+) and then were plated onto gelatin-coated dish. Cultured cells were observed daily using an inverted light microscope to determine the day of contraction onset and total duration of continuous contractile activity for each contracting focus. This frequency was compared with the results of DMSO not treated P-mES02 group (4-/4-) and mES01 groups (4-/4+ or 4-/4-). For confirm the generation of cardiomyocytes, beating cell masses were treated with trypsin-EDTA, dispersed cells were plated onto glass coverslips and incubated for 48 h. Attached cells were fixed using 4% paraformaldehyde and incubated with specific antibodies (Abs) to detect cardiomyocytes (anti-sarcomeric ? -actinin Ab, 1 : 100; anti-cardiac troponin I Ab, 1 : 2000) for 1 h. And the cells were finally treated with FITC or TRITC labelled 2nd Abs, respectively, then they were examined under fluorescence microscopy. Results: Rhythmically contracting areas in mES01 or P-mES02 cells were firstly appeared at 9 or 10 days after EBs plating, respectively. The highest cumulative frequency of beating EBs was not different in both treatment groups (mES01 and P-mES02, 4-/4+) with the results of 61.3 % at 13 days and 69.8% at 15 days, respectively. Also, the contracting duration of individual beating EBs was different from minimal 7 days to maximal 53 days. However, DMSO not treated groups (mES01 and P-mES02, 4-/4-) also had contracting characteristics although their frequency was a few compared to those of DMSO treated groups (6.0% and 4.0%). Cells recovered from the spontaneously contracting areas within EBs in both treated groups were stained positively with muscle specific anti-sarcomeric ? -actinin Ab and cardiac specific anti-cardiac troponin I Ab. Conclusion: This study demonstrated that the P-mES02 cell-derived cardiomyocytes displayed similarly structural properties to mES01 cell-derived cardiomyocytes and that the DMSO treatment enhanced the cardiomyocytes differentiation in vitro.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.18
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• pp.7849-7855
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• 2014

Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.