• Asian-Australasian Journal of Animal Sciences
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• v.21 no.8
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• pp.1116-1126
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• 2008

The objective of this study was to investigate the expression and localization of heat shock protein 70 (Hsp70) and its mRNA in the heart, liver, and kidney of acutely heat-stressed broilers at various stressing times. Male AA broilers (n = 100) were randomly divided into 5 groups of 20 birds per group. After 30 d of adaptive feeding at ambient temperature, 80 experimental broilers were suddenly heat stressed by increasing the environmental temperature from $22{\pm}1^{\circ}C$ to $37{\pm}1^{\circ}C$. The 4 groups were heat stressed for 2, 3, 5, and 10 h, respectively. The localizations of Hsp70 protein and mRNA, determined by immunohistochemical staining and in situ hybridization, respectively, were demonstrated to be tissue dependent, implying that different tissues have differential sensibilities to heat stress. Intense Hsp70 staining was identified in the vascular endothelial cell of heart, liver and kidney, suggesting an association between expression of Hsp70 in vascular endothelial cell and functional recovery of blood vessels after heat shock treatment. Ante-mortem heat stress had a significant effect on the expression of Hsp70 protein and mRNA. The quantitation of Hsp70 protein and mRNA were both time and tissue dependent. During the exposure to heat stress, the heart, liver and kidney of broiler chickens exhibited increased amounts of Hsp70 protein and mRNA. The expression of hsp70 mRNA in the heart, liver and kidney of heat-stressed broilers increased significantly and attained the highest level after a 2-h exposure to elevated temperatures. However, significant elevations in Hsp70 protein occurred after 2, 5, and 3 h of heat stressing, respectively, indicating that the stress-induced responses vary among different tissues.

• The Korean Journal of Cytopathology
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• v.8 no.2
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• pp.129-134
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• 1997

To distinguish reactive mesothelial cells from malignant cells in body fluid, we applied silver staining of nucleolar organizer regions(AgNORs) to ethanol fixed cytologic preparations. Fifty aspirated samples of benign(22 cases) and malignant(26 cases) body fluids were studied using the one step silver staining method. Two cytologically atypical samples were also included in the study. In malignant cases the mean AgNOR count was $3.56{\pm}0.81$, while in benign cases the mean AgNOR count was $2.02{\pm}0.33$. The difference of AgNOR counts between these two groups were statistically significant(p<0.001). The mean of atypical cases was 2.91. Both were diagnosed as malignant in follow-up cytology. In malignant effusions, there is statistically significant difference in AgNOR counts between cells forming complex papillae or clusters and singly scattered cells(p<0.05), $3.29{\pm}0.95\;and\;3.83{\pm}0.55$, respectively. We concluded that AgNOR count appears to be useful as a diagnostic tool especially when the cytologic differentiation is difficult.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.21
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• pp.9071-9075
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• 2014

Background: microRNAs are small non-coding RNA that control gene expression by mRNA degradation or translational inhibition. These molecules are known to play essential roles in many biological and physiological processes. miR-205 may be differentially expressed in head and neck cancers; however, there are conflicting data and localization of expression has yet to be determined. Materials and Methods: miR-205 expression was investigated in 48 cases of inflammatory, benign and malignant tumor tissue array of the neck, oronasopharynx, larynx and salivary glands by Locked Nucleic Acid in situ hybridization (LNA-ISH) technology. Results: miR-205 expression was significantly differentially expressed across all of the inflammatory, benign and malignant tumor tissues of the neck. A significant increase in miR-205 staining intensity (p<0.05) was observed from inflammation to benign and malignant tumors in head and neck tissue array, suggesting that miR-205 could be a biomarker to differentiate between cancer and non-cancer tissues. Conclusions: LNA-ISH revealed that miR-205 exhibited significant differential cytoplasmic and nuclear staining among inflammation, benign and malignant tumors of head and neck. miR-205 was not only exclusively expressed in squamous epithelial malignancy. This study offers information and a basis for a comprehensive study of the role of miR-205 that may be useful as a biomarker and/or therapeutic target in head and neck tumors.

• Korean journal of dermatology
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• v.56 no.10
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• pp.631-635
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• 2018

Microvenular hemangioma (MVH) is a rare acquired benign vascular neoplasm, which presents commonly as a solitary purple-to-red nodule or plaque measuring approximately 10 mm in diameter. MVH occurs primarily on the extremities or the trunk. Most lesions are solitary, and multiple lesions are rare. Histopathological features of MVH include numerous, scattered, thin and irregularly branching small vessels in the dermis and endothelial cells without atypia. Owing to similarities in clinical morphology and histopathological features, MVH may often be indistinguishable from the early patch stage of Kaposi sarcoma. Immunohistochemical (IHC) analysis helps differentiate between the 2 diseases. The results of IHC tests in patients with MVH show positive staining for CD31 and smooth muscle actin and typically, negative staining for the human herpes virus 8 antigen. We report a rare case of multiple MVH clinically mimicking the early patch stage of Kaposi sarcoma in a 63-year-old woman who presented with a 3-year history of slowly growing, compressible, soft, bluish-purple macules and plaques on the trunk and right arm.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.26 no.3
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• pp.36-53
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• 2013

Objectives : The purpose of this study is to investigate the effects of DOGO phreatic water containing sulphur on Atopic Dermatitis in NC/Nga mouse. Methods : We made DOGO phreatic water removed sulphur using Twin Alternating Sulfate Eater. After making atopic dermatitis caused by sensitizing NC/Nga mouse to DNCB(dinitrochlorobenzene), we made mouse swim in tanks each filled with distilled water, tap water, DOGO phreatic water(contain sulphur), DOGO phreatic water(remove sulphur) for 30minutes everyday. 3weeks later, we analyzed skin clinical score, total IgE levels(by ELISA), WBC differential counting(Neutrophils, Monocytes), absolute cell number of $Neutrophil^+Gr-1^+$, CCR3 mRNA expressions(by Real-time PCR), IL-4, IFN-${\gamma}$ production levels(by ELISA), histologic test(by H&E staining, toluidine blue staining). Results : The results of making NC/Nga mouse induced atopic dermatitis swim in tanks filled with DOGO phreatic water(contain sulphur) are as follows. 1. Skin clinical scores were decreased significantly in comparison to control group. 2. Total IgG levels were decreased significantly in comparison to control group. 3. WBC differential counting(Neutrophils, Monocytes) were decreased significantly in c.mparison to control group. 4. Absolute cell number of $Neutrophil^+Gr-1^+$ were decreased significantly in comparison to control group. 5. CCR3 mRNA expressions were decreased significantly in comparison to control group. 6. IL-4, IFN-${\gamma}$ production levels were decreased significantly in comparison to control group. 7. The epithelial tissue thickness, leucocytes infiltration, erythema, edema, excoriation, scaling, mast cells infiltrations in dorsal skin were decreased in comparison to control group. Conclusions : These results indicate that DOGO phreatic water(contain sulphur) can be used for helping treat atopic dermatitis.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.1
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• pp.1-7
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• 2004

연구 목적: 본 연구의 목적은 마우스에서 배양액의 용량이 배반포 배 형성과 세포수에 미치는 영향을 조사하기 위하여 실시하였다. 연구 재료 및 방법: $3{\sim}4$주령 ICR 암마우스에게 48시간 간격으로 5 IU PMSG와 hCG 주사 후 (hCG 주사 후 수컷과 동숙) $46{\sim}50$시간에 난관으로부터 총 138개의 2-세포기 배를 회수하여 2 ml (group I) 또는 $50{\mu}l$ (group II)의 배양액 (Dulbecco's Modified Eagle Medium + 20% human follicular fluid)에서 72시간 동안 배반포기까지 배양하였다. 배반포 배는 zona-intact (ZiB)와 zona-escape (ZeB)로 등급을 구분하고 나서, propidium iodide와 bisbenzimide를 이용한 differential staining 방법으로 염색하여 평균 세포수, 내세포괴(ICM) 세포수, 영양배엽(TE) 세포수, 총 세포수에 대한 ICM의 비율 (%ICM) 및 ICM:TE 비율을 조사하였다. 결과에 대한 유의성 검정은 $X^2$ test와 t-test를 이용하였으며, p<0.05일 때 통계적인 차이가 있는 것으로 하였다. 결 과: Group I과 II에서, 총 배반포 ($62.3{\pm}20.7%$ vs. $63.8{\pm}22.9%$), ZiB ($31.9{\pm}24.0%$ vs. $30.4{\pm}18.2%$)와 ZeB 형성율 ($30.4{\pm}20.8%$ vs. $33.3{\pm}22.3%$)은 차이가 없었다. 87개의 배반포 배를 염색 시도하였는데, 명확하게 differential staining된 41개의 배반포 배만을 대상으로 세포수를 조사하였다. 평균 세포수 ($61.6{\pm}19.5$ vs. $63.7{\pm}26.8$), ICM 세포수 ($13.0{\pm}10.6$ vs. $12.8{\pm}10.5$), TE 세포수 ($49.0{\pm}19.0$ vs. $47.8{\pm}18.7$), %ICM ($21.0{\pm}12.6%$ vs. $21.1{\pm}13.2%$) 및 ICM:TE 비율 (1:$3.77{\pm}4.9$ vs. 1:$3.72{\pm}4.8$)에서도 group I과 II에서 차이가 없었다. 결 론: 마우스에서 배 발생 능력의 척도로 쓰이는 배반포 배 형성율 배반포 배의 등급 세포수 및 %ICM 등이 20% 난포액을 첨가한 MEM 배양액의 용량에 따라 영향을 받지 않았다.

• Korean Journal of Microbiology
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• v.39 no.1
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• pp.27-35
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• 2003

No currently available methods to monitor pathogenic protozoa, Cryptosporidium and Giardia in environmental water come close to acceptable sensitivity, specificity and reproducibility, and so it has to be accompanied by thorough quality control and performance evaluation to credibly predict the distribution of them. We collected surface water samples from the Han River and spiked our prepared (oo)cysts, determined Matrix Spike recoveries using USEPA Method 1623 and considered what factors influence MS recovery and validity. As a result, average 46% of spiked oocysts and 60% of spike cysts were recovered, but repetitive sampling and statistical approach seemed to be necessary to determine the environmental pollution level of two protozoa as their variation coefficients was so much as 35oio and 26%. And MS recoveries with two acid dissociations during immunomagnetic separation were improved more 10% than that with one dissociations and the use of spiked suspension enumerated by flow cytometry instead of manual preparation enhanced the validity and reliability in spiking tests. Because fluorescence characteristics of (oo)cysts stained on well slides with FITC-labeled monoclonal antibodies and DAPI was not always same, well Elides from spiked field samples were helpful to evaluate the performance of staining. We found many (oo)cyst-like objects with typical fluorescence, not (co)cysts, from the Han River water samples, and then it was concluded that nuclei staining by DAPI (4',6-diamidino-2-phenylindole) and examination by Differential Interference Contrast Microscope should be critical for valid identification.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.4
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• pp.237-243
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• 2002

Objective : The aim of this study was to evaluate the influence of different media on blastulation, mean cell number, percentage of inner cell mass (ICM) of total cells and ICM : trophectoderm (TE) ratio in mice. Materials and methods: A total 552 two cell embryos were retrieved from ICR female mice (4 weeks old) at 48 hr after hCG injection (mated just after hCG injection) and cultured in MEM (n=276) or TCM (n=276) supplemented with 20% hFF. The grading of blastocysts from zona-intact (ZiB) to -escape (hatching and hatched, ZeB) was performed at 72 hours after culture. Total, TE and ICM cell numbers were analyzed by differential staining of blastocyst. Statistical analysis was performed using t-test with SigmaPlot-2001. P-values < 0.05 were accepted as statistically significant. Results: The blastulation rate in MEM ($64.9{\pm}4.95%$) was significantly higher (p=0.0031) than that in TCM ($57.2{\pm}5.22%$). No differences were found in the number of ZiB and ZeB between MEM ($31.9{\pm}2.62$, $33.0{\pm}4.58%$), and TCM ($27.2{\pm}4.28$, $30.1{\pm}4.58%$). A total 314 blastocysts (MEM=166; TCM=148) were stained differentially. Mean cell number of blastocysts was significantly higher (p=0.0002) in TCM ($73.1{\pm}3.3$) than in MEM ($61.7{\pm}2.5$). Differential staining was successfully performed in 155 blastocysts (MEM=77; TCM=78). The percentage of ICM was significantly higher in MEM than in TCM ($20.9{\pm}1.3$ vs. $17.1{\pm}1.2%$, p=0.0281). The ICM : TE ratio was higher in TCM than in MEM (1 : $4.85{\pm}0.68$ vs. 1 : $3.78{\pm}0.78$, NS). Conclusion: These results show that MEM increase the blastocyst formation and percentage of ICM of total cells comparing with TCM in mice.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.27 no.4
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• pp.141-157
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• 2014

Objectives : The purpose of this study is to investigate the effects of DOGO phreatic water containing germanium on Atopic Dermatitis in NC/Nga mouse. Methods : We made DOGO phreatic water added germanium. After making atopic dermatitis caused by sensitizing NC/Nga mouse to DNCB(dinitrochlorobenzene), we made mouse swim in tanks each filled with distilled water, tap water, DOGO phreatic water, DOGO phreatic water(added germanium) for 30minutes everyday. 3weeks later, we analyzed skin clinical score, total IgE levels(by ELISA), WBC differential counting(Neutrophils, Monocytes), absolute cell number of $Neutrophil^+Gr-1^+$, CCR3 mRNA expressions(by Real-time PCR), IL-4, IFN-${\gamma}$ production levels(by ELISA), histologic test(by H&E staining, toluidine blue staining). Results : The results of making NC/Nga mouse induced atopic dermatitis swim in tanks filled with DOGO phreatic water(contain germanium) are as follows. 1. Skin clinical scores were decreased significantly in comparison to control group. 2. Total IgG levels were decreased significantly in comparison to control group. 3. WBC differential counting(Neutrophils, Monocytes) were decreased significantly in comparison to control group. 4. Absolute cell number of $Neutrophil^+Gr-1^+$ were decreased significantly in comparison to control group. 5. CCR3 mRNA expressions were decreased significantly in comparison to control group. 6. IL-4, IFN-${\gamma}$ production levels were decreased significantly in comparison to control group. 7. The epithelial tissue thickness, leucocytes infiltration, erythema, edema, excoriation, scaling, mast cells infiltrations in dorsal skin were decreased in comparison to control group. Conclusions : These results indicate that DOGO phreatic water(contain germanium) can be used for helping treat atopic dermatitis.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.68-68
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• 2001

Blastocyst formation, consisting of the inner cell mass (ICM) and trophectoderm (TE), is the first differentiation process during embryonic development in mammals. It has been hypothesized that the proportion of ICM to TE in the blastocyst may be crucial for subsequent developmental competence of early embryos, which it may be expressed as a sensitive indicator for evaluating in vitro systems. In this study ICM/total cell ratio of nuclear transfer (NT) embryos was compared with IVF-derived and in vivo embryos. Somatic cell nuclei obtained from a fetus at Day 40 of gestation were transferred into the enucleated oocyte and then cultured in NCSU 23 medium for 6 days as previously described (Koo et al., Biol. Reprod. 2000; 63:986-992). ICM and TE cells of blastocysts were determined by using a differential staining method (Han et al., Biol. Reprod. 1999; 60:1110-1113). Development rate (9.8$\pm$2.5%, 23/225) to the blastocyst stage of NT embryos was lower than IVF embryos (23.8$\pm$2.7%, 53/223). Thus, a difference was detected in the in vitro developmental rate to blastocyst stage between NT and IVF-derived embryos (P<0.05). In the next experiment, we investigated ICM and TE nuclei to assess the quality of blastocysts that produced by NT, IVF and in vivo, respectively. NT blastocysts (27.6$\pm$8.3) showed a smaller total cell number than IVF-derived (42.6$\pm$17.4) and in vivo embryos (283.9$\pm$103.5) (P<0.05). Ratios of ICM/total cells in NT, IVF and in vivo blastocysts were 15.1$\pm$ 18.6% (n=56), 12.3$\pm$9.2% (n=57) and 30.4$\pm$6.8% (n=40), respectively. Individual blastocysts for the ratio of ICM/total cells were assigned to 3 groups (I; <20%, II; 20 to 40% and III;>40%). As the results, most in vivo blastocysts (97.5%, 39/40) were distributed into group II while most NT (78.6%, 44/56) and IVF-derived blastocysts (82.5%, 47/57) were allocated to group I. Thus, our data show that NT or IVF-derived embryos have aberrant morphology during early development in vitro systems, suggesting that these anomalies may result in developmental failures of the NT embryos to term.