• Animal Bioscience
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• v.34 no.6
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• pp.993-1005
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• 2021

Objective: Zearalenone (ZEA) has estrogen-like effects. Our previous study has shown that ZEA (0.5 to 1.5 mg/kg) could induce abnormal uterine proliferation through transforming growth factor signaling pathway. To further study the other regulatory networks of uterine hypertrophy caused by ZEA, the potential mechanism of ZEA on porcine endometrial epithelial cells (PECs) was explored by the Illumina Hiseq 2000 sequencing system. Methods: The PECs were treated with ZEA at 0 (ZEA0), 5 (ZEA5), 20 (ZEA20), and 80 (ZEA80) µmol/L for 24 h. The collected cells were subjected to cell cycle, RNA-seq, real-time quantitative polymerase chain reaction, immunofluorescence, and western blot analysis. Results: The proportion of cells in the S and G2 phases decreased (p<0.05), but the proportion of cells in the G1 phase increased (p<0.05) in the ZEA80 treatment. Data analysis revealed that the expression of Wnt pathway-related genes, estrogen-related genes, and mitogen-activated protein kinase pathway-related genes increased (p<0.05), but the expression of genetic stability genes decreased (p<0.05) with increasing ZEA concentrations. The relative mRNA and protein expression of WNT1, β-catenin, glycogen synthase kinase 3β (GSK-3β) were increased (p<0.05) with ZEA increasing, while the relative mRNA and protein expression of cyclin D1 (CCND1) was decreased (p<0.05). Moreover, our immunofluorescence results indicate that β-catenin accumulated around the nucleus from the cell membrane and cytoplasm with increasing ZEA concentrations. Conclusion: In summary, ZEA can activate the Wnt/β-catenin signaling pathway by up-regulating WNT1 and β-catenin expression, to promote the proliferation and development of PECs. At the same time, the up-regulation of GSK-3β and down-regulation of CCND1, as well as the mRNA expression of other pathway related genes indicated that other potential effects of ZEA on the uterine development need further study.

• Journal of Life Science
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• v.31 no.8
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• pp.755-760
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• 2021

Swine diarrhea is a livestock disease that causes huge economic losses to pig farms. In general, diarrhea occurs because of the proliferation of pathogenic Escherichia coli (E. coli). The toxins produced by the proliferated E. coli cause edema in pigs. Although the proliferation of these coliforms can be prevented by using a vaccine, the vaccines containing chemically produced dead bacteria are not very effective, making it difficult to control the proliferation of E. coli. Therefore, there is a need to develop new, more effective vaccines. In this study, we prepared killed F4+ and F18ab+ E. coli, which induce diarrhea and edema in pigs, using the extracts of immune-induced silkworms containing antimicrobial peptides and examined their availability as a killed-bacteria vaccine. First, the antimicrobial activity analysis of the prepared immune-induced silkworm extract was conducted using the radial diffusion assay. The results showed high activity against both F4+ and F18ab+ E. coli. The production efficiency of E. coli dead cells was determined using the colony-counting method. The concentration of the E. coli dead cells was the highest (50 mg/ml) when treated at 4℃. In addition, the analysis of the prepared dead cells using a transmission electron microscope confirmed that E. coli leaked out of the cytoplasm and the cell membrane remained intact. Therefore, F4+ and F18ab+ E. coli produced using immune-induced silkworms extract are considered to be highly available as bacterial ghost vaccines that can help prevent swine diarrhea and the resulting edema.

• The Korea Journal of Herbology
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• v.33 no.4
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• pp.59-67
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• 2018

Objectives : Ojeoksan, originally recorded in an ancient Korean medicinal book named "Donguibogam" and has been used for the treatment of circulation disorder of blood which was called blood accumulation (血積) in Korean medicine. Therefore, this study was carried out to investigate the beneficial effect of OJS on vascular inflammation in HUVECs. Methods : We evaluated the effect of OJS on the expression of cell adhesion molecules and protective role in HUVEC stimulated by TNF-${\alpha}$ by using Western blot. Results : Pretreatment with OJS decreased the adhesion of HL-60 cells to TNF-${\alpha}$-induced HUVEC. OJS suppressed TNF-${\alpha}$-induced expression level of cell adhesion molecules such as intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1(VCAM-1), and endothelial cell selectin (E-selectin). Moreover, OJS significantly decreased TNF-${\alpha}$-induced production of intracellular reactive oxygen species (ROS); and inhibited the phosphorylation of $I{\kappa}B-{\alpha}$ in the cytoplasm compared to the experimental group. Pretreatment with OJS inhibited the trans-location of NF-${\kappa}B$ p65 to the nucleus. OJS also inhibited phosphorylation of MAPKs compared to the experimental group. OJS significantly increased the protein expression of Nrf2 and HO-1. Conclusions : Ojeoksan has a protective effect on vascular inflammation, and might be a potential therapeutic agent for early atherosclerosis.

• Nutrition Research and Practice
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• v.16 no.3
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• pp.330-343
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• 2022

BACKGROUND/OBJECTIVES: Zanthoxylum schinifolium is traditionally used as a spice for cooking in East Asian countries. This study was undertaken to evaluate the anti-proliferative potential of ethanol extracts of Z. schinifolium leaves (EEZS) against human bladder cancer T24 cells. MATERIALS/METHODS: Subsequent to measuring the cytotoxicity of EEZS, the anti-cancer activity was measured by assessing apoptosis induction, reactive oxygen species (ROS) generation, and mitochondrial membrane potential (MMP). In addition, we determined the underlying mechanism of EEZS-induced apoptosis through various assays, including Western blot analysis. RESULTS: EEZS treatment concentration-dependently inhibited T24 cell survival, which is associated with apoptosis induction. Exposure to EEZS induced the expression of Fas and Fas-ligand, activated caspases, and subsequently resulted to cleavage of poly (ADP-ribose) polymerase. EEZS also enhanced the expression of cytochrome c in the cytoplasm by suppressing MMP, following increase in the ratio of Bax:Bcl-2 expression and truncation of Bid. However, EEZS-mediated growth inhibition and apoptosis were significantly diminished by a pan-caspase inhibitor. Moreover, EEZS inhibited activation of the phosphoinositide 3-kinase (PI3K)/Akt pathway, and the apoptosis-inducing potential of EEZS was promoted in the presence of PI3K/Akt inhibitor. In addition, EEZS enhanced the production of ROS, whereas N-acetyl cysteine (NAC), a ROS scavenger, markedly suppressed growth inhibition and inactivation of the PI3K/Akt signaling pathway induced by EEZS. Furthermore, NAC significantly attenuated the EEZS-induced apoptosis and reduction of cell viability. CONCLUSIONS: Taken together, our results indicate that exposure to EEZS exhibits anti-cancer activity in T24 bladder cancer cells through ROS-dependent induction of apoptosis and inactivation of the PI3K/Akt signaling pathway.

• Journal of Applied Biological Chemistry
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• v.65 no.4
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• pp.239-245
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• 2022

Although normal activation of platelets is important in the process of hemostasis, excessive or abnormal activation of platelets can lead to cardiovascular diseases. Therefore, the discovery of novel substances capable of regulating or inhibiting platelet activation may be helpful in the prevention and treatment of cardiovascular diseases. Artemether is a derivative of artemisinin, known as an active ingredient of Artemisia annua, which has been reported to be effective in treating malaria, and is known to function through antioxidant and metabolic enzyme inhibition. However, the role of artemether in platelet activation and aggregation and the mechanism of action of artemether in collagen-induced human platelets are not known until now. This study investigated the effects of artemether on platelet activation and thrombus formation induced by collagen. As a result, cAMP level was significantly increased by artemether, and VASP and IP3R, substrates of cAMP-dependent kinase, were phosphorylated. IP3R phosphorylation by Artemether inhibited Ca²⁺ recruitment into the cytoplasm, and phosphorylated VASP inhibited fibrinogen binding by inactivating αIIb/β3 located on the platelet membrane. Consequently, artemether inhibited thrombin-induced fibrin clot formation. Therefore, we propose that artemether can act as an effective prophylactic and therapeutic agent for cardiovascular diseases caused by excessive platelet activation and thrombus formation.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.78-78
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• 2002

The role of heat shock proteins in shielding organism from environmental stress is illustrated by the large-scale synthesis of these protein by the organism studied to date. However, recent evidence also suggests an important role for heat shock protein in fertilization and early development of mammalian embryos. Effects of elevated in vitro temperature on in vitro produced bovine embryos were analysed in order to determine its impact on the expression of heat shock protein 70 (HSP70) by control and frozen-thawed after in vitro fertilization (IVF) or nuclear transfer (NT). The objective of this study was to assess the developmental potential in vitro produced embryos with using of the various containers and examined expression and localization of heat shock protein 70 after it's frozen -thawed. For the vitrification, in vitro produced embryos at 2 cell, 8 cell and blastocysts stage after IVF and NT were exposed the ethylene glycol 5.5 M freezing solution (EG 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min, and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid, cryo-loop. However, survival rates by straw were relatively lower than other containers. Only, nuclear transferred embryos survived by using cryo-loop. After IVF or NT, in vitro matured bovine embryos 2 cell, 8 cell and blastocysts subjected to control and thawed conditions were analysed by semiquantitive reverse transcription polymerase chain reaction methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNA were higher thawed embryos than control embryos. Immunocytochemistry used to localization the hsp70 protein in embryos. Two, 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some embryos exposed frozen-thawed. However, under control condition, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform in distribution.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.30 no.4
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• pp.431-435
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• 1985

Nine hybrid rices crossed between Korean cytoplasmic-genetic male sterile rices having the WA cytoplasm and the Korean restorer lines or varieties, HR1619A/Nampungbyeo, HR1619A/Gayabyeo, HR1619A/Line234, TongilA/Nampungbyeo, TongilA/Cheongcheongbyeo, Suwon 296A/Line 209, Suwon 296A/Line 237, Suwon 296A/Line 252 and Line 201A/Line 234, and their parents were grown at Yeungnam University in 1984. The rough rice yield of the hybrids Line 201A/Line 234, TongilA/Nampungbyeo and HR1619A/Nampungbyeo were 939, 927 and 900 Kg/10a respectively. The heterosis(F$_1$/Midparent) of the above three hybrids was 40%, 20% and 19%, the heterobeltiosis(F$_1$/Better parent) was 36%, 17% and 10%, and the standard heterosis (F$_1$/Standard variety, Cheongcheongbyeo) was 19%, 17% and 14% respectively. The hybrids HR1619A/ Gayabyeo and Suwon 296A/Line 237 showed negative heterosis in grain yield. Significant heterobeltiosis for grain number per panicle was found while less or no heterobeltiosis was observed in panicle number per hill, 1000-grain weight and grain fertility. The bacterial leaf blight disease reaction of the hybrids tested was almost the same as that of one parent at least. The amylose content of the hybrids was medium to low the same as their parents. The protein content and alkali digestion value of the hybrids were almost the same as their parents.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.223-231
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• 2001

The success of nuclear transplantation with mammalian oocytes depends critically on the potential of oocytes activation, which mainly caused to prevent the re-accumulation of maturation promoting factor (MPF). This study was conducted to compare the effect of combined treatment of lonomycin with a Hl-histone kinase inhibitor (dimethylaminopurine, DMAP) or cdc2 kinase inhibitor (sodium pyrophosphate, SPP) on activation of bovine oocytes. In vitro matured bovine oocytes with the first polar body (PB) and dense cytoplasm were assigned to 3 experimental groups. For activation treatment, oocytcs were exposed to 5 $\mu$M lonomycin for 5 min (Group 1), and followed by 1.9 mM dimethylaminopurine (DMAP) for 3 h (Group 2) or followed by 2 mM sodium pyrophosphate (SPP) for 3 h (Group 3). The activation effects in the three treatments and the control group (untreated) were judged by the extrusion of the second PB and formation of a pronucleus (PN). Differences among groups were analysed using one-way ANOVA after arc-sine transformation of proportional data. All three treatments led to high activation rates (90% to 95%), with significant difference from the control. However, the extrusion of the second PB and the rate of PN formation differed remarkably among treatments. In Group I and 3, about 95% of the oocytes had extruded the second polar body, but one PN had formed in a higher proportion of oocytes in Group 3 than in Group 1 (90% vs. 5%). In experiment 2, the rates of cleavage and development into blastocysts in Group 1 were significantly lower than those of Group 2 and 3 (8.7% and 0% vs. 50.5% and 11.6%, and 44.6% and 7.2%, respectively, P<0.05). In experiment 3, ~80% of parthenotes in Group 1 were developed with haploid chromosomal sets. However, when ionomycin was followed immediately by DMAP (Group 2). only 20% of parthenotes were haploid. In Group 3, combined treatment with ionomycin and SPP, the appearance of abnormal chromosomal tracts was significantly (P〈0.05) reduced and the proportion of haploid parthenotes was increased to 85% (17/20) than in Group 2. These results demonstrate that SPP acted as a cdc2 kinase inhibitor and formed the haploidy in oocyte activation. Thus, the present study suggests that cdc2 kinase inhibitor, such as sodium pyrophosphate, may have an effective role in oocyte activation for the production of cloned embryos/animals by nuclear transplantation.

• The Korean Journal of Zoology
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• v.15 no.3
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• pp.111-131
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• 1972

Fine structures of retina of an ommatidium in dragonfly, having eyes of closed rhabdom type, were studied under light and electron microscopes. The ommatidia consisted of eight retinular cells distributed in a circular pattern and the retinular layer in turn can be divided into three sublayers according to the number of cells in the retina. Each retinular cell has different starting points in the retina and the length of retinular cells is varied greatly; the length of one distal retinular cell shows one half of that of others. In the middle layer, three proximal retinular cells interconnect the adjacent two rhabdoms which are triangular in the appearence of the cross section which in turn consisted of tubular, parallel and lamellated microvilli. The rhabdom is formed by three rhabdomeres, each of which is separated by $120^{\circ}$ between them, but they can be distinguished into two parts according to electron density. Around the outer part of microvilli composing rhabdom, electron density was much less than the inner part of the structure. The microvilli of the inner part appear to be connected to the cytoplasm of retinular cells. Rough endoplsmic reticulum with enlarged cisternae runs through the vacuoles in the outer part of distal retinular cells. Abundant mitochondria concentrated in the vicinity of rhabdom are found at the central part of the retinular cells, while in the area of immediate vicinity of the rhabdom, prominent vacuoles are observed. Above the rhabdom of an ommtidium stands a crystalline cone which is consisted of four cone cells arranged radially along the axis. The crystalline cone is surrounded by cells containing pigment granules. The outermost photoreceptor element of an ommatidium is corneal lens.

• The Korean Journal of Zoology
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• v.22 no.3
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• pp.103-114
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• 1979

The authors observed the ultrastructure of the granular glands in the amphibian skin with an electron microscope. The specimens from the experimental animals (Bombina orientalis, Bufo bufo gargarizans, Rana nigromaculata and Rana rugosa) were fixed in 2.5% glutaraldehyde-paraformaldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 1% osmium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with a LKB-ultramicrotome. the ultrathin sections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follws: 1. The granular gland in the amphibian skin consisted of the glandular epithelial and the myoepithelial cells. 2. The epithelial cells of the granular gland in the amphibian skin consisted of the dark cells but the light cells were also observed in that of Bombina orientalis. 3. The granular glands of the amphibian skin were in holocrine fashion. 4. The nuclei of the epithelial cells of the amphibian cutaneous granular glands were round or oval and showed small and large inforldings of nuclear envelope. Heterochromatins were mainly distributed near the nuclear envelope. Mitochondria were mainly distributed in the perinuclear portion and rough-surfaced endoplasmic reticulums were developed in the cytoplasm but smooth-surfaced endoplasmic reticulums were not well developed. 5. Secretory granules were round or oval and electron-dense and less electron-dense granules were observed. 6. The authors infer that the differences in electron density of the secretory granules in the granular glands of the amphibian skin are due to difference in the concentrations of secretory substances as related to the processes of its formation, and that those chemical components are identical.