• Development and Reproduction
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• v.22 no.2
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• pp.155-163
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• 2018

This study aimed to investigate changes in the activity and mRNA expression of plasminogen activators (PAs) induced by $17{\beta}$-estradiol ($E_2$), human chorionic gonadotropin (hCG), and interleukin-$1{\beta}$ ($IL-1{\beta}$) in porcine endometrial cells. Endometrial cells were isolated from the epithelium and cultured to 80% confluence. They were then treated for 24 h with $E_2$ (0.2, 2, 20, and 200 ng/mL), $IL-1{\beta}$ (0.1, 1, 10, and 100 ng/mL), and hCG (0.5, 1, 1.5 and 2 IU/mL). mRNA expressions of urokinase-type (uPA) and tissue-type (tPA) PAs were analyzed using reverse transcription PCR, and activities were measured using a PA activity assay. mRNA expressions of uPA and tPA increased with $E_2$ treatment; however, this was not significant. Similarly, treatment with hCG did not influence the mRNA expressions of PAs. Interestingly, treatment with 0.1 ng/mL $IL-1{\beta}$ significantly reduced the mRNA expression of uPA, but did not affect that of tPA. Treatment with 2, 20, and 200 ng/mL $E_2$ increased PA activity compared with the control group; treatment with 0.1 and 1 ng/mL $IL-1{\beta}$ significantly increased PA activity compared with the other $IL-1{\beta}$ treatment groups, whereas treatment with 10 and 100 ng/mL $IL-1{\beta}$ decreased. Treatment with 2 IU/mL hCG increased PA activity compared with the other treatment groups, although there were no significant differences between the hCG and control groups. In conclusion, the activity and mRNA expression of PAs were differently regulated by the hormone/cytokine and its concentration in porcine endometrial cells. Therefore, understanding PA regulatory mechanisms may help to improve the reproductive potential of domestic animals.

• Applied Microscopy
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• v.31 no.2
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• pp.109-116
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• 2001

Safflower has been cultivated in Korea and thought to have excellent effects on bone in oriental medicine and folk remedy and has been taken for a long time. Safflower is thought to be helpful for the development and sustenance of bones according to the result of recent assay of its components. Otherwise, any reliable experimental data have not been suggested so far. We have carried out this study to examine the prophylactic effects of safflower-seed-powder on the prevention of osteoporosis induced by the ovariectomy 12 Weeks-old Sprague-Dawley rats weighing about 230 g was kept in the experimental condition and used in this study. Animals were taken 0.3 g of safflower-seed-powder once a day for 1, 3, 5, and 7 weeks after ovariectomizing both ovaries and observed the fine structure of tibia. Tissues were fixed with traditional SEM preparation methods and decalcified for 10 hours with 10% nitric acid and dehydration, drying, and gold-coating were followed by the routine procedures and observed with scanning electron microscope (Hitachi, S-450). Loss of bone was started just after ovariectomy and thickness of bone from the medullary cavity to the compact bone was reduced and the extension of medullary cavity was serious in the control group of 7 weeks. Experimental groups taken safflower-seed-powder showed similar findings from 1 week to 7 weeks. These results suggest that the safflower-seed-powder is thought to be efficient for the prevention of osteoporosis owing to the deficiency of female sex hormone.

• Journal of Veterinary Clinics
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• v.20 no.3
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• pp.376-383
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• 2003

To establish the differential diagnosis and treatment method in bovine ovarian cysts, specially ovarian cysts with corpus luteum, serum progesterone concentration, rectal palpation and ultrasonography for measuring the cystic wall thickness and diameter of cyst and corpus luteum were investigated from 1,188 dairy cows with ovarian cysts. The plasma progesterone concentrations were 0.3$\pm$0.4 (mean$\pm$SD) ng/ml in 629 cows with follicular cysts, 3.7$\pm$1.1 ng/ml in 431 cows with luteal cysts, and 3.8$\pm$1.2 ng/ml in 128 cows with coexist of ovarian cysts and corpus luteum, respectively. The cystic wall thickness by ultrasonography were 1.6$\pm$0.4 mm in 629 cows with follicular cysts, 4.2$\pm$1.5 mm in 431 cows with luteal cysts, and 1.6$\pm$0.6 mm in 128 cows with coexist of ovarian cysts and corpus luteum, respectively. The days from initial treatment to insemination in follicular cysts were 28.1$\pm$6.9 days in treatment of GnRH alone, 15.9$\pm$2.9 days in combination of GnRH and dinoprost, and 15.1$\pm$3.1 days in combination of GnRH and cloprostenol. The percentages of cows conceived within 100 days after initial treatment were 61 %, 68% and 73% in treatment of GnRH alone, combination of GnRH and dinoprost, and combination of GnRH and cloprostenol, respectively. The days from initial treatment to insemination in luteal cysts were 3.8$\pm$0.6 days in treatment of dinoprost alone and 3.8$\pm$0.7 in cloprostenol alone. The percentages of cows conceived within 100 days after initial treatment were 69.5% and 68.5% in treatment of dinoprost and cloprostenol, respectively. The days from initial treatment to insemination in coexist of cysts and corpus luteum were 3.7$\pm$0.7 days in treatment of dinoprost alone and 3.8$\pm$0.6 in cloprostenol alone. The percentages of cows conceived within 100 days after initial treatment were 87% and 84% in treatment of dinoprost and cloprostenol, respectively. These results suggest that the best choice for treatment agents in ovarian cysts were combination of GnRH and PGF$_2$$\alpha$ in follicular cysts, and the PGF$_2$$\alpha$ in luteal cysts and in coexist of cysts and corpus luteum, respectively. In conclusion, it is suggest that ultrasonography is useful diagnostic tool for diagnosis and selection of treatment remedy in cystic ovaries of bovine.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.21 no.4
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• pp.332-344
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• 1999

Cytokines are hormone-like proteins which mediate and regulate inflammatory and immune responses. Interleukin-6 (IL-6) is involved in the final differentiation of B cells into antibody-producing cells. Interleukin-8 (IL-8) is a neutrophil chemotactic factor that plays an important role in the recruitment of neutrophil to inflammatory loci. Inflammatory mediators by cells in the gingiva have been implicated in the initiation and progression of periodontitis and oral infection. The purpose of this study was conducted to investigate the effect of lipopolysaccharide (LPS), staphylococcus enterotoxin B (SEB) on production of IL-6 and IL-8 by human gingival and facial dermal fibroblasts. Primary cultured human gingival and facial dermal fibroblasts were incubated with LPS (0.01, 0.1, $1.0{\mu}g/ml$), SEB (0.01, 0.1, $1.0{\mu}g/ml$) or LPS $(0.1{\mu}g/ml)$ plus SEB $(0.1{\mu}g/ml)$. Culture supernatants were collected at 24, 48, and 72 hrs and assessed for IL-6 and IL-8 production by enzyme-linked immunosorbent assay. IL-6 production in gingival fibroblasts stimulated with LPS was higher than that with SEB. IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-6 production in facial dermal fibroblasts was increased only by stimulation with a high concentration of LPS $(1.0{\mu}g/ml)$. Its production in facial dermal fibroblasts by exposure with SEB was decreased in comparison with control, nontreated cells. Therefore, gingival fibroblasts showed higher sensitivity than facial dermal fibroblasts in response to low concentration of LPS. Also, IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-8 production in gingival fibroblasts was enhanced greatly only by stimulation of high concentration of LPS $(1.0{\mu}g/ml)$. That by exposure with SEB was increased only in 24 hrs cultivation. IL-8 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-8 production in facial dermal fibroblasts was decreased by LPS and increased only in 48 hrs cultivation by SEB. IL-8 production by double exposure with LPS plus SEB was enhanced only in 48 hrs cultivation in comparison with single exposure of LPS or SEB. therefore, IL-6 and IL-8 production were released at various quantities according to bacterial toxin applied and site of fibroblast harvested. These results suggest that gingival fibroblasts may be concerned with IL-6 and IL-8 related inflammatory response more than facial dermal fibroblasts.

• Journal of Embryo Transfer
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• v.28 no.4
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• pp.315-318
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• 2013

This study was conducted to investigate the changes of hormone levels of serum progesterone (P4) and estradiol-$17{\beta}$ (E2) in sows of Landrace (L), Yorkshire (Y) and F1 (L ${\times}$ Y) (respectively n=3) with excellent ability, and to provide a baseline data for improving reproductive performance. In this experiment, the sows at the age of 12 months or more were used. The sows were fed by two way methods, one is conventional methods and the other is 3 days-flushing feed before estrus. Each pig's blood was collected in 3, 6, 9, 12, 15 and 18 days after the estrus for the analyses of P4 and E2. Serum was separated by centrifugation for 15 min. with 3,000 rpm. Progesterone and estradiol-$17{\beta}$ were measured by immunochemical assay (ELIZA test). In conventional feeding, serum progesterone levels were significantly (p<0.01) higher in F1 than in L and Y. No significant differences in P4 concentrations were seen between the L and Y of sows. Serum E2 levels were similar the serum progesterone levels. In the case of flushing feed, the tendency of hormonal changes were similar to conventional methods. But almost of hormonal levels were a little higher than that of conventional methods. P4 level of L and Y in flushing feed were significantly different (p<0.01). Serum E2 level of Y in flushing feed was significantly different among the breeds (p<0.01). These results were similar to the tendency of hormonal changes in general sows and moreover, flushing feed is known to develop the swine production, these results proved the fact of the methods. And these results suggested that more studies about hormonal changes in sows according to seasonal and nutritional factors should be needed.

• Journal of the Korean Applied Science and Technology
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• v.34 no.4
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• pp.995-1003
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• 2017

Whitening and anti-oxidant effects were observed in order to investigate the biological activations of Salvia plebeia herb ethanol extracts. No toxicity was found in both B16F10 melanoma cells and Raw 264.7 cells exposed to Salvia plebeia herb ethanol extracts for 48 hour. The extracts showed significant antioxidant activity in cell-free and cell-cultured system. In the DPPH radical assay, it removed dose-dependently DPPH radicals and showed 77.6% at $100{\mu}g/mL$. In the cells, it also significantly removed silica-induced ROS generation and LPS-induced NO production in a dose dependent manner. Using L-DOPA and L-tyrosine as a substrate, tyrosinase activity was inhibited using Salvia plebeia herb ethanol extracts in a dose-dependent manner. The supression occurred to be in the B16F10 melanoma cells, where dose-dependently inhibited Salvia plebeia herb ethanol extracts of $1{\mu}g/M$ ${\alpha}$-melanocyte stimulated hormone-induced melanin production and the inhibitory effect was 30.7% at a concentration of $100{\mu}g/mL$. This suggests that the Salvia plebeia herb ethanol extracts are usable for cosmetic product developments for anti-oxidant and whitening effects.

• Korean Journal of Plant Resources
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• v.31 no.4
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• pp.312-321
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• 2018

Angelica tenuissima, also known as Ligusticum tenuissimum, is classified as a food-related plant and has been used as traditional medicines treating headache and anemia in Asia. However, its anti-melanogenic effect has not been reported in detail. When the extract of Angelica tenuissima (ATE) was prepared by the extraction with 70% EtOH at $80^{\circ}C$ (final yield = 22%), the contents of decursin and Z-ligustilide in ATE were determined 0.06% and 8.43%, respectively. Total flavonoid and phenolic content in mg ATE were $5.52{\pm}0.07{\mu}g$ quercetin equivalents and $237.27{\pm}13.24{\mu}g$ gallic acid equivalents, respectively. Antioxidant capacity of ATE determined by DPPH and ABTS assay was increased with a dose dependent manner up to $1000{\mu}g/m{\ell}$. The amount of melanin synthesis followed by ${\alpha}-melanocyte$ stimulating hormone on B16F10 cells were significantly reduced in the presence of ATE (250 to $1000{\mu}g/m{\ell}$, p<0.05). ATE (125 to $1000{\mu}g/m{\ell}$, p<0.05) suppressed the tyrosinase activity but did not show any significant effect on ${\alpha}-glucosidase$ activity at the same condition. Taken together, ATE possesses tyrosinase inhibitory potential with significant antioxidant capacities. These effects of ATE might be involved in suppression of melanin synthesis, at least, in B16F10 cells. The anti-melanogenic potential of ATE will provide an insight into developing a new skin whitening product.

• The Korea Journal of Herbology
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• v.32 no.2
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• pp.57-64
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• 2017

Objectives : Yam bean (Pachyrhizus erosus) possess various nutrients, it has been widely used as traditional cosmetic material in Indonesia. The aim of this study was to investigate the anti-oxidant activity and the anti-melanogenic effect of Yambean (Pachyrhizus erosus) extract and its fractions. Methods : The anti-oxidant activity of yam bean extract assessed based on total polyphenol, flavonoid contents, DPPH and ABTS radical scavenging assay. To evaluate anti-melanogenic effects and cytotoxicity of Yambean extract and its fractions, B16F10 melanoma cell was used. Results : In results, total polyphenol content of yam bean water extract (YW) and Yambean 70% ethanol extract (YE) were $1.18{\pm}0.03mg/g$ (mg of gallic acid/g of sample), $1.16{\pm}0.01mg/g$. Total flavonoid contents of YW, YE were $3.55{\pm}0.06mg/g$ (mg of naringin/g of sample), $1.78{\pm}0.03mg/g$. Moreover, YE scavenged DPPH and ABTS effectively in $4mg/m{\ell}$ compared to YW. Cytotoxicity of YE and its fractions in B16F10 melanoma cell was measured using MTT assays. It had no cytotoxicity up to $500{\mu}g/m{\ell}$. Melanin accumulation in B16F10 melanoma cell was induced using alpha-melanocyte stimulating hormone (${\alpha}-MSH$) and 3-isobutyl-1-methylxanthine (IBMX). B16F10 melanoma cell treated with $10-500{\mu}g/m{\ell}$ YE and hexane, ethyl acetate, butanol, $H_2O$ fractions for 24h. Non treated B16F10 melanoma cell (Control) markedly increased melanin contents. In contrast, YE ethylacetate fraction effectively suppressed melanin accumulation in a dose-dependent manner. Conclusion : In conclusion, these results suggest that Yambean extract has the potential as a cosmetic material which possess anti-oxidant and anti-melanogenic activities.

• The Korea Journal of Herbology
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• v.33 no.4
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• pp.77-85
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• 2018

Objective : This study was intended to examine the effects of water extract of Prunellae Spica (PS), which is a herb with 'cold' nature based on hot and cold theory of traditional Korean medicine. Methods : Hyperthyroidism was induced in SD rats by LT4 (0.5 mg/kg, i.p.) daily for four weeks. After 2 weeks of LT4 injection, rats were divided randomly into four groups; normal, LT4-induced hyperthyroid control, PS extract (500 mg/kg, p.o.)-treated group, and propylthiouracil (PTU, 10 mg/kg, s.c.)-treated positive group. After 2 weeks of drug treatment, all rats were sacrificed and harvested blood samples and thyroid tissues. The changes of body weight, food and water intake, and body temperature were measured weekly. Serological markers were analyzed in sera using an enzyme-based assay, and thyroid tissues were stained with Hematoxylin & Eosin (H&E). Brain and dorsal root ganglion (DRG) tissues were isolated and analyzed the expression of transient receptor potential (TRP) channels by Western blot. Results : PS extract administration attenuated the loss of body weight and the increase of body temperature in LT4-induced hyperthyroidism rats. PS extract increased the level of thyroid stimulating hormone (TSH) and decreased tiiodothyronine (T3) and tetraiodothyronine (T4). In action mechanism, PS extract regulated the expression of transient receptor potential channel subfamily V member 1 (TRPV1) and transient Receptor Potential channel subfamily M member 8 (TRPM8), the thermoregulators. Conclusion : To conclude, PS extract can improve the symptoms of hyperthyroidism through regulation of the thyroid hormones imbalance and thermoregulation via TRP channels.

• Animal Bioscience
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• v.35 no.12
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• pp.1850-1859
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• 2022

Objective: The quantitative reverse transcription polymerase chain reaction (qPCR) is the most accurate and reliable technique for analysis of gene expression. Endogenous reference genes (RGs) have been used to normalize qPCR data, although their expression may vary in different tissues and experimental conditions. Verification of the stability of RGs in selected samples is a prerequisite for reliable results. Therefore, we attempted to identify the most stable RGs in the hypothalamic-pituitary-gonadal (HPG) axis in sows. Methods: The cycle threshold values of nine commonly used RGs (18S, HPRT1, GAPDH, RPL4, PPIA, B2M, YWHAZ, ACTB, and SDHA) from HPG axis-related tissues in the domestic sows in the different stages of estrus cycle were analyzed using two RG-finding programs, geNorm and Normfinder, to rank the stability of the pool of RGs. In addition, the effect of the most and least stable RGs was examined by normalization of the target gene, gonadotropin-releasing hormone (GnRH), in the hypothalamus. Results: PPIA, HPRT1, and YWHAZ were the most stable RGs in the HPG axis-related tissues in sows regardless of the stages of estrus cycle. In contrast, traditional RGs, including 18S and ACTB, were found to be the least stable under these experimental conditions. In particular, in the normalization of GnRH expression in the hypothalamus against several stable RGs, PPIA, HPRT1, and YWHAZ, could generate significant (p<0.05) elevation of GnRH in the preovulatory phase compared to the luteal phase, but the traditional RGs with the least stability (18S and ACTB) did not show a significant difference between groups. Conclusion: These results indicate the importance of verifying RG stability prior to commencing research and may contribute to experimental design in the field of animal reproductive physiology as reference data.