• Journal of the Korea Academia-Industrial cooperation Society
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• v.15 no.4
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• pp.2189-2198
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• 2014

Paraquat (PQ) is a very effective and widely used herbicide that was commercially introduced in 1962. In this study, instead of using antioxidants like in the past, to inhibit the formation of PQ-induced ROS, we attempted to reduce the oxygen concentration by using non-lethal hypoxia therapy. Therefore, we studied the toxicity of PQ in vivo, analyzed the major effects of ROS on the targeted lung tissue and compared the results with the gross histological changes after the cell protective effect of non-lethal hypoxia therapy. In vivo studies demonstrated that low-concentration oxygen therapy (i.e., 10-12% oxygen) in rats administered with PQ was associated with a higher survival rate than in rats that received only PQ. In vivo non-lethal hypoxia treatment showed better survival and less lung tissue damage. Using a hypoxic/anaerobic incubator with integrated multifaceted molecular analysis, including MDA assay, glutathione assay, and SOD assay, we established an optimal, significantly reduced in vivo non-lethal hypoxia treatment by exploiting the PQ-induced cytotoxicity responses.

• Journal of Embryo Transfer
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• v.29 no.1
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• pp.13-19
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• 2014

This study was carried out to study the survival rate of thawed Hanwoo embryos frozen by the slow-rate freezing or the cryotop vitrification method. Hanwoo cumulus-oocyte complexes were recovered from ovaries at a slaughter house, matured for 20~22 hours, fertilized with Hanwoo semen for 5~6 hours, and cultured for 7~9 days in $38.5^{\circ}C$, 5% $CO_2$ incubator. For freezing, Day 7~9 blastocysts were collected. Embryos for the slow-rate freezing were equilibrated in 1.8 M ethylene glycol (EG) with Dulbecco's phosphate-buffered saline (D-PBS). Programmable cell freezer was precooled down to $-7^{\circ}C$, and the straw was seeded during 8 minutes-holding time, and was cooled to $-35^{\circ}C$ at the cooling rate of $0.3^{\circ}C/min$, and then was plunged and stored in liquid nitrogen. Embryos for the cryotop vitrification were treated in TCM199 with 0.5 M sucrose, 16% EG, 16% dimethylsulfoxide (DMSO). Embryos were then loaded individually onto cryotop and plunged directly into liquid nitrogen. The survival rates of embryos frozen by these two freezing methods were evaluated at 12 to 24h post-thawing. The survival rates of frozen/thawed Hanwoo embryos by the cryotop vitrification method ($56.86{\pm}26.53%$) were slightly higher than those by the slow-rate freezing method ($55.07{\pm}26.43%$) with no significant difference. Using the cryotop vitrification and the slow-rate freezing of Hanwoo blastocysts on Day 7 following in-vitro fertilization (IVF) treatment, the survival rates of frozen/thawed Hanwoo embryos were $72.65{\pm}18.3%$ and $79.06{\pm}17.8%$, respectively. The survival rates by the cryotop vitrification were higher than those by the slow-rate freezing on both Day 8 and 9 with significantly higher survival rate on Day 9 (p<0.05). Using the cryotop vitrification and the slow-rate freezing of Hanwoo embryos to compare between three different blastocyst stages, the survival rates of the blastocyst stage embryos were $66.22{\pm}18.8%$ and $45.76{\pm}12.8%$, respectively with higher survival rate by the vitrification method (p<0.05). And the survival rate of expanded blastocysts was higher than those of early blastocysts and blastocysts in two freezing methods with significantly higher survival rate by the slow-rate freezing method (p<0.05).

• The korean journal of orthodontics
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• v.26 no.4
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• pp.449-454
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• 1996

The propose of this study was to evaluate the difference of cellular activity dependent on intermittent compressive force by determining the alkaline phosphatase activity. Alkaline phosphatase activity was measured on control and experimental groups every 24, 48, 72hours. Experimental groups consisted of continous and intermittent compressive group which were compressed by $300gm/cm^2$ of diaphram pump. The intermittent compressive group was connected by timer which was worked on 10 minutes an off 10 minutes. The results were as follows; 1. The alkaline phosphatase activity between control and experimental groups showed not significant difference at compressed 24 hours. 2. The alkaline phosphatase activity of experimental groups were more increased than control group at compressed 48 hours. 3. The alkaline phosphatase activity of intermittent compressive group showed significant increased to control group. Whereby continuous compressive group showed not significant difference to control at 72 hours. 4. The alkaline phosphatase activity of intermittent compressive group were stringly increased than continuous compressive groups. 5. Between experimental groups and control group no other morphologic changes were detected by microscopic findings.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.419-424
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• 1998

The present study was carried out to examine the efficiency of cloning of transgenic embryos by nuclear transfer(NT) using gene-injected rabbit embryos. The rabbit embryos at pronuclear stage were microinjected with methallothionein-human growth hormone(MT-hGH) gene and cultured to 8- and 16-cell in TCM-199 containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% $CO_2$incubator. The recipient oocytes were collected from the oviducts 14~16 h after hCG injection. The oocytes were enucleated and activated with 5$\mu$M ionomycin and 2mM 6-dimethylaminopurine. Blastomeres form gene-injected embryos were transferred into the enucleated oocytes by micromanipulation. The nuclear transplant oocytes were electrofused and co-cultured with rabbit oviductal cells. Following 120 h of culture, blastocysts were prepared for gene analysis by polymerase chain reaction(PCR). In previous experiment, the rate of gene-positive embryos detected by the nested PCR analysis was significantly decreased while developing to blastocyst(25%)(Kang et al., 1998). The fusion rate of gene-injected blastomeres was significantly(P＜0.05) lower than non-injected blastomeres(66% vs 80%). However, the NT embryos that were derived from gene-injected donor embryos did not differ from control embryos in development to the blastocyst stage(39% vs 31%). Of the 43 NT blastocysts developed from the gene-injected donor embryos, twelve(28%) were positive for the injected DNA. The results indicate that NT with gene-injected embryos can be successfully used for cloning and multiplication of transgenic embryos, furthermore applicable to improvement of transgenic animal production.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.14 no.12
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• pp.6413-6419
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• 2013

This study examined the irradiation effect of high energy x-rays on the hatching process of fertilized eggs, particularly with regard to malformation and blood cells change. The experimental groups were five day old fertilized eggs irradiated with x rays at doses of 5, 7.5, 10 Gyusing alinear accelerator. The control group was not irradiated. After three weeks, hatched chicks were sacrificed and examined for blood sampling. The survival rate of the x-ray irradiated groups were significantly lower than that of the control group (46.7vs 80%). The malformation rate of the experimental groups was60%, whereas no congenital malformations were observed in the control group. The experimental groups had a significantly higher malformation rate. The types of malformation were left wing defect, proptosis, microcephaly, cervical spine, and feet anomaly. The incidence of malformation increased with increasing radiation dose. The white blood cell count of control group and eachexperimental groups (5 and 7.5 Gy) were 87.64, 100.76 and 81.42 ($10^3/{\mu}L$), respectively. X-ray irradiation of 5 day old chick embryos increased the rate of death and malformation significantly.The malformations were estimated to have occurred by chromosomal abnormalities. Further genetic studies will be needed to confirm the correlation between high energy X-rays and the cause of malformations.

• The Journal of the Korean bone and joint tumor society
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• v.7 no.1
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• pp.1-12
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• 2001

Purpose : To elucidate possibility of local chemotherapy from adraimycin-impregnated bone cement. Materials and Methods : Authors used 4 kinds of bone cements, Palcos R, LVC, CMW 3, Simplex P for this experimental model, included 2.5mg, 5mg, 25mg of adriamycin, respectively. We compared the differences of eluted-adriamycin concentrations between the cylindrical shape and the flat shape of bone cements, between ddH2O, 0.45% saline, 0.9% saline, and 3% saline as one of environmental conditions. Osteosarcoma cell line, Saos-2 were cultured under $37^{\circ}C$, 5% $CO_2$ in the humidified incubator with three different concentrations of adriamycinimpregnated bone cements and cellular toxicity of adriamycin eluted from bone cement was analysed according to MTT assay. Results : Authors noticed the flat shape of bone cement eluted more concentrations of adriamycin than the cyclindrical shape, bone cement immersed in 3% saline, more than 0.9% or 0.45% saline. Concentrations of adriamycin eluted from CMW 3 or Simplex R were more than Palacos R or LVC. Saos-2 were cultured with 2.5mg, 5mg, 25mg of adriamycin-impregnated bone cement, respectively, and their cellular toxicity were 95%, 98%, 99%, each. Conclusion : Adriamycin-impregnated bone cement can be one of anticancer-drug delivery sytems as possible local chemotherapy.

• Journal of Periodontal and Implant Science
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• v.31 no.1
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• pp.179-193
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• 2001

previous studies have demonstrated an increase in bone mass and density with use of bisphosphonate in osteoporosis. This agent acts as an inhibitor of osteoclastic activity and results in increase of net osteoblastic activity. The purpose of the present study was to examine the effect of the bisphosphonate on osteoblastic activity of the human periodontal ligament cells in vitro. Periodontal ligament cells were primarily obtained from extracted healthy third molars. Cells of 4th to 6th passage were cultured in Dulbecco's modified Eagle's medium containing alendronate sodium or etidronate disodium at the concentration of $10^{-12}{\sim}10^{-6}mol/L$ in 5% $Co_2$ incubator at $37^{\circ}C$. Cell count and MTT assay for cellular activity were done at 2 to 7 days of culture. Alkaline phosphatase activity at 4 to 7 days of culture and formation of mineralized nodules at 28 days of culture with addition of $50{\mu}g/m{\ell}$ ascorbic acid, 10 nM${\beta}-glycerophosphate$, $10^{-7}M$ dexamethasone were evaluated. 1. Alendronate sodium Compared to the control, the proliferation of periodontal ligament cells was generally increased and the cellular activity was maintained at 2 days of culture and generally decreased at 7 days of culture. Alkaline phosphatase activity of periodontal ligament cells was inceased and the formation of mineralized nodules by periodontal ligament cells was enhanced compared to the control. 2. Etidronate disodium The proliferation of periodontal ligament cells was increased at 2 days of culture and decreased or maintained at 7 days of culture. Compared to the control, the cellular activity of periodontal ligament cells was generally decreased. Alkaline phosphatase activity of peridontal ligament cells was increased and the formation of mineralized nodules by periodontal ligament cells was enhanced compared to the control. These results suggest that alendronate sodium and etidronate disodium may have a potential effect on osteoblastic lineage of periodontal ligament cells, distinct from their inhibitory action on osteoclasts and could contribute to enhance periodontal regeneration and alveolar bone regeneration.

• Journal of Periodontal and Implant Science
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• v.24 no.3
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• pp.661-670
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• 1994

It is well known that the application of dressings after periodontal surgery have benefits to provide the comforts to patient and to promote the healing process with action of bleeding control and temporary stabilization for the operated mobile teeth. But until recently the relationship between periodontal dressings and cells which are composed of periodontium has not been clear. The purpose of this study was to evaluate the cytotoxic effect of soluble extracts from the four different kinds of periodontal dressings, two of them were eugenol type (K.H.pack, Wondrpak) and the others were non-eugenol type (Coe-pak, Periocare), on the human gingival fibroblasts in vitro. Human gingival fibroblasts were primarily cultured from gingiva around third molar during the extraction for preventive purposes. Extracts solution were prepared with culture medium by means of imersing the consistent size of periodontal dressing made from plastic mold. Cell were inoculated into the 24 well plate with $3\;{\times}\;10^4\;cells/well$ of medium at $37\;^{\circ}C$, 100% of humidity, 5% of $CO_2$, incubator for 24 hours. After discard of the supernatant of medium, those cells were cultured with original, 1/2, 1/5, 1/10 diluted soluble extract for 24, 48 and 72 hours, and counted the number of cells using the hemocytometer at each designed time and concentration. Also, the cytotoxic effect of soluble extract was measured by Wataha's MTT assay method. In briefly, cells were inoculated and cultured into 96 well culture plate with $2\;{\times}\;10^4\;cells/well$ for 24 hours. Soluble extracts were applied to cultured cells and incubated for 48 hours at same condition. $50\;{\mu}l$ of MTT solution and DMSO were added into each well for the detection of absorbance with ELISA reader. The measured data were calculated by value of colorimetric assay for survival rate. The results were as follows ; In the case of eugenol type of dressing, original, 1/2 and 1/5 diluted extracts of K.H.pack showed very low survival rate. And original extract of Wondrpak showed strong cytotoxic effect and 1/2 diluted extract showed moderate cytotoxic effect. In the case of Non-eugenol type of dressings, only original extract of Coe-pak revealed strong cytotoxic effect and Periocare had little cytotoxic effect. It is concluded that eugenol type of dressings showed more cytotoxic effect than non-eugenol types. This study suggest that use of non-eugenol dressings after periodontal surgery is recommended.

• Applied Microscopy
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• v.38 no.3
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• pp.259-264
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• 2008

The stabilized $ClO_2$ gas has been used for many years by the food industry as a strong oxidizing and sanitizing agent that has broad and high biocidal effectiveness. Therefore, "stabilized $ClO_2$" gas may be used in fields of disinfectant and sterilization. But, there have been few studies on the decomposition-inhibition effect of stabilized $ClO_2$ gas with passage of time. The main purpose of this study was to examine the decomposition-inhibition effect of stabilized $ClO_2$ gas and the morphological change of kidney by measuring of the light and electron microscope. Sprague-Dawley (SD) rats weighting from 230 gm to 250 gm were used as experimental animals. Under ether anesthesia, the right kidney of rat was obtained. Put each sample in $37^{\circ}C$ and humidity $80{\pm}5%$ incubator, we obtained each sample after 0 day, 1 day, 2 days, 3 days, 4 days and 5 days. We proceeded the observation of light and electron microscope. The results obtained in this study reveal that stabilized $ClO_2$ gas is an effective decomposition inhibitor until 2 days that was conducted at $37^{\circ}C$ and humidity $80{\pm}5%$ conditions.

• Korean Journal of Animal Reproduction
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• v.14 no.3
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• pp.183-190
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• 1990

These studies were carried out ot investigate the effects of estrous cow serum(ECS), fetal calf serum(FCS), bovine follicular fluid(BFF) and matured cumulus cell(MCC) on in vitro maturation and fertilization of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluid from the visible follicles of diameter 3-5mm. The follicular oocytes were cultured in TCM-199 medium containing hormones, FCS, ECS, BFF and MCC for 24~48 hrs. in a incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 18$^{\circ}C$20 hrs. with motile capacitated sperm in the TCF(Tyrode calcium-free) solution containing 200$\mu\textrm{g}$/ml of heparin. The results obtained in these experiments were summarized as follows : 1. The oocytes classified as "A, B, C, D and Degenerative" depending morphological integrity and those were 61.4%, 12.1%, 19.2%, 4.2% and 3.0% of the total oocytes harvested, respectively. The maturation and fertilization rate of the A, B, C class follicular oocytes, cultured in the TCM-199 medium supplemented with 10% FCS were 89.1%, 78.0%, 52.6% and 78.1%, 33.3%, respectively. 2. The maturation and fertilization rate of the follicular oocytes cultured in TCM-199 medium supplemented with 5%~20% ECS and FCS were 74.0%~80.6, 26.2%~30.0% and 71.7%~76.9%, 51.9%~58.0%, and those values were higher the supplement of ECS than FCS. 3. The maturation rate(68.0%~64.6%) and fertilization rate(59.6%~60.4%) of follicular oocytes cultured in TCM-199 medium supplemented with 10% FCS and 20~30% BFF were higher than those of follicular oocytes cultured TCM-199 medium supplemented with 10% FCS and 10% and 50% BFF. 4. The maturation rate(76.5%) and fertilization rate(61.7%) of follicular oocytes cultured in TCM-199 medium supplemented with 10% FCS and 1$\times$106/ml cumulus cells were higher than those of follicular oocytes cultured in TCM-199 medium supplemented with 10% FCS and 1$\times$104~5/ml and 1$\times$108/ml cumulus cells.lus cells.