• Clinical and Experimental Reproductive Medicine
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• v.34 no.4
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• pp.275-283
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• 2007

Objective: This study was performed to investigate the expressions of endometriosis related genes in shed endometrial tissues from menstrual blood of patients with or without endometriosis. Methods: The shed endometrial tissues were collected on 2$^{nd}$ or 3$^{rd}$ day of menstrual cycle with Wallace catheter in patients with endometriosis (n=16) and without endometriosis (n=26). The mRNA expressions of twelve kinds of endometriosis related genes were compared between two groups using semi-quantitative RT-PCR. Results: The collected shed endometrium was confirmed by histological observation. Expressions of telomerase, c-kit and aromatase mRNA were not detected by RT-PCR in shed endometrial tissues. The mRNA expressions of apoptosis related genes (fas, fas ligand, bcl-2, bax), stem cell factor, estrogen receptor-$\alpha$/$\alpha$, endometriosis protein-I and secretory leukocyte protease inhibitor gene were similar between shed endometrial tissues with endometriosis and without endometriosis. Conclusion: We could not find the difference of mRNA expressions of tested endometriosis related genes between shed endometrial tissues with or without endometriosis by semi-quantitative RT-PCR analysis. It may be related to the dynamical changes of gene expressions in the endometrium with menstrual cycle.

• Polymer(Korea)
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• v.27 no.1
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• pp.9-16
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• 2003

Ipriflavone (IP) stimulates proliferation and differentiation of osteoblast and also enhances calcitonin secretion in the presence of estrogen. Poly(lactide-co-glycolide) (PLCA) due to its controllable biodegradability and relatively good biocompatibility is one of the most significant candidates for the study of drug controlled release system. In this study, IP-loaded PLGA microspheres (MSs) was prepared by using conventional O/W solvent evaporation method. The size of MSs was in the range of 5~200 $mu extrm{m}$. The morphology of MSs was characterized by SEM. And, in vitro release amounts of IP were analyzed by HPLC. The highest encapsulation efficiency were obtained by using gelatin and polyvinyl alcohol (PVA) as emulsifiers. The morphology, size distribution, and in vitro release pattern of MSs were changed by several preparation parameters such as molecular weights (8, 20, 33 and 90 kg/mol), polymer concentrations (2.5, 5, 10 and 20%), emulsifier types (PVA, gelatin and Tween 80), initial drug loading amount (5, 10, 20 and 30%) and stirring speed (250, 500 and 1000 rpm). The release of IP in vitro was more prolonged over 30 days, with close to zero-order pattern by controlling the preparation parameters. The physicochemical properties of IP-loaded PLGA MSs were investigated by XRD and DSC.

• Korean Journal of Food Science and Technology
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• v.32 no.1
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• pp.25-30
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• 2000

Phytoestrogen has been used as a suppliment of estrogen in order to treat osteoporosis. The representative phytoestrogens, isoflavones, are daidzein, genistein and formononetin which were present highly in our traditional soybean foods. The quantitative analysis of the isoflavone was done with a high performance liquid chromatography(HPLC) using a UV/VIS detector for the contents of the isoflavones in Astragali Radix, soybean sprouts, bean-curd(Tofu), soybean, soybean oil, pea, kidney pea, black bean(Yak-kong), soybean sauce(Ganjang). soybean paste(Doenjang), and fermented soybean(Maejoo). The content of free daidzein in soybean sprouts was $43.49{\pm}3.41\;mg/kg$$ which was much higher than that in soybean, $14.52{\pm}0.58\;mg/kg$, although total daidzein of fermented soybean was lower than that in soybean (Table 2. P<0.01). The amounts of free genistein in soybean sprouts, fermented soybean. and soybean paste were $27.63{\pm}1.66\;mg/kg,\;291.52{\pm}6.81\;mg/kg,\;and\;18.75{\pm}1.33\;mg/kg$, respectively. The level of free formononetin in soybean paste was the highest among the soybean products (P<0.01). The content of formononetin in Aatragali Radix, $9629.73{\pm}0.57\;{\mu}g/kg$, was about 160 times higher than that in soybean. Thus Korean traditional soybean products, black bean(Yak-Kong) and Astragali Radix can be a good choice of phytoestrogens.

• Korean Journal of Food Science and Technology
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• v.44 no.1
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• pp.82-88
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• 2012

Chrysanthemum indicum L. (Asteraceae) is a traditional herbal medicine that has been used for the treatment of inflammation, hypertension, and respiratory diseases due to its strong antagonistic activity against inflammatory cytokines. The effects of Chrysanthemum indicum L. Extract (CIE) for increasing cell growth, alkaline phosphatase (ALP) activity, and collagen content were totally inhibited, suggesting that the effect of CIE might be partly involved with estrogen activity. Furthermore, the protective effects of CIE on the response of osteoblasts to oxidative stress were evaluated. Osteoblastic MC3T3-E1 cells were incubated with hydrogen peroxide and/or CIE, and markers of osteoblast function and oxidative damage were examined. CIE significantly increased cell survival, ALP activity, and calcium deposition, and decreased the production of Reactive Oxygen Species (ROS) and Tumor Necrosis Factor-${\alpha}$ (TNF-${\alpha}$) in osteoblasts. Taken together, these results indicate that the enhancement of osteoblast function by CIE may prevent osteoporosis and inflammatory bone diseases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.6
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• pp.833-839
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• 2005

This study was conducted to demonstrate whether dietary Ca and the soy isoflavones supplementation could reduce the bone loss associated with estrogen deficiency. Nine week-old female rats (SD) were ovariectomized and then fed on diet of low $(0.1\%)$ or normal $(0.5\%)$ Ca supplemented with soy isoflavones (80 and 160 ppm) for 6 weeks. The ovariectomized and sham-operated rats showed normal serum Ca and P levels, and dietary Ca and soy isoflavones did not changed them. The serum alkaline phosphatase activity was increased in all ovariectomized rats, especially in the rats fed low Ca diet regardless of isoflavone supplementation. The serum tartrate-resistance acid phosphatase was not significantly different among the ovariectomized rats and were not changed by dietary Ca and isoflavones. Breaking force of femur was higher in the rats fed the nomral Ca diets and not statistically changed by soy isoflavone supplementation. Femoral and lumbar Ca and P contents decreased in the ovariectomized rats and the soy isoflavones-80 ppm supplementation significantly enhanced bone minerals, but the soy isoflavones-160 ppm supplementation did not. Dietary Ca increased lumbar Ca and P contents. The results of this study have suggested that the soy isoflavones 80 ppm supplementation could be sufficient to prevent bone loss in ovariectomized rats and normal Ca supplementation could enhance the effect of soy isoflavones on bone protection.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.3
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• pp.521-526
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• 2002

A number of experimental and epidemiological studies have implicated that antiestrogenic effects of estrogen-like compounds in legumes and plant seeds are responsible for lowering breast cancer risk in human. However, few studies have been conducted to illustrate the possible chemopreventive effects of Korean traditional food materials. This study was performed to determine the cytotoxic and apoptotic effects of yellow soybeans, black soybeans and brown rice extracts on hormone-dependent and hormone-independent human breast cancer cells. Methanol-or acetone-soluble fractions of soybeans or brown rice were incubated with hormone-dependent cells (MCF-7) or hormone-independent cells (MDA-MB-231). Cell cytotoxicity was measured by MTT assay at 24, 48 and 72 hrs of incubation. Apoptotic effects of these extracts toward breast cancer cells were also determined at 48 hrs of incubation by measuring DNA fragmentation. Results indicated that the acetone-soluble fraction of brown rice exerted strongest cytotoxic effect on MCF-7 ceIls, although other fractions also reduced the number of viable MCF-7 cells after 48 hrs of incubation. Both acetone and methanol soluble fractions of all samples exerted a significant cytotoxicity towards MDA-MB-231 cells after 24 hrs of incubation, and acetone and methanol soluble fractions of brown rice were especially effective in these cells. At 48 hrs of incubation, methanol fractions of all three samples induced apopotosis of MDA-MB-231 cells. These results indicate methaol or acetone soluble fractions of yellow soybeans, black soybeans and brown rice induce cytotoxicity in both hormone-dependent and hormone-independent breast cancer cells. Therefore, possible mechanisms of cell cytotoxicity do not necessarily include antiestrogenic effects of soybean or brown rice extract. A possible anticarcinogenic effect of brown rice methanol-soluble fraction may mediated through their apoptotic effect. Further studies are requried to elucidate responsible compounds and mechanisms involved in observed anticarcinogenesis.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.207-219
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• 2014

Three-dimensional (3D) culture system is useful technique for study of in vivo environment and it was used various experiments. This study was investigated to establish of embryo co-culture system and changes of PAs activity in 3D cultured endometrial cells of pigs. In results, growth of stromal cells into gel matrix were detected only with endometrial and myometrial cells. The most rapid growth of stromal cells were confirmed in $2.5{\times}10^5cells/ml$ and gel matrix containing 15% FBS. Expression of urokinase-PA (uPA) after treatment of hCG (0.5, 1.0, 1.5 and 2.0 IU/ml) were higher than without hCG, but, there are not significant difference among the treatment. On the other hand, expression of uPA after treatment of $IL-1{\beta}$ (0.1, 1, 10 and 100 ng/ml) were higher than without $IL-1{\beta}$, but, there are not significant difference. Expression of uPA after treatment of estrogen (0.2, 2, 20 and 200 ng/ml) were not difference, but PA activity was significantly decreased (p<0.05). Blastocyst was producing in PZM-3 medium containing FBS and endometrial cells were grown in PZM-3 medium. When embryos development with cultured endometrial cells, cleavage rates were not significant difference and blastocyst were not produced in co-culture with stromal cells and 3D culture system. 3D culture system had similar activity to in vivo tissue and these features are very useful for study of in vivo physiology. Nevertheless 3D culture system was not proper in embryo co-culture system. Therefore, we suggest that 3D culture system with embryo co-culture need continuous research.

• Journal of Animal Science and Technology
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• v.56 no.7
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• pp.26.1-26.10
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• 2014

Background: Endocrine disruptors are exogenous substance, interfere with the endocrine system, and disrupt hormonal functions. However, the effect of endocrine disruptors in different species has not yet been elucidated. Therefore, we investigated the possible effects of $17{\beta}$-estradiol (E2), progesterone (P4), genistein (GEN) and 4-tert-octylphenol (OP), on capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. In this in vitro trial, spermatozoa were incubated with $0.001-100{\mu}M$ of each chemical either 15 or 30 min and then assessed capacitation status using chlortetracycline staining. Results: E2 significantly increased capacitation and the acrosome reaction after 30 min, while the acrosome reaction after 15 min incubation in mouse spermatozoa. Simultaneously, capacitation and the acrosome reaction were induced after 15 and 30 min incubation in porcine spermatozoa, respectively. Capacitation was increased in porcine spermatozoa after 15 min incubation at the lowest concentration, while the acrosome reaction was increased in mouse spermatozoa after 30 min (P < 0.05). E2 significantly increased the acrosome reaction in porcine spermatozoa, but only at the highest concentration examined (P < 0.05). P4 significantly increased the acrosome reaction in bovine and mouse spermatozoa treated for 15 min (P < 0.05). The same treatment significantly increased capacitation in porcine spermatozoa (P < 0.05). P4 significantly increased capacitation in mouse spermatozoa treated for 30 min (P < 0.05). GEN significantly increased the acrosome reaction in porcine spermatozoa treated for 15 and 30 min and in mouse spermatozoa treated for 30 min (P < 0.05). OP significantly increased the acrosome reaction in mouse spermatozoa after 15 min (P < 0.05). Besides, when spermatozoa were incubated for 30 min, capacitation and the acrosome reaction were higher than 15 min incubation in E2 or GEN. Furthermore, the responsiveness of bovine, mouse and porcine spermatozoa to each chemical differed. Conclusions: In conclusion, all chemicals studied effectively increased capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. Also we found that both E2 and P4 were more potent than environmental estrogens in altering sperm function. Porcine and mouse spermatozoa were more responsive than bovine spermatozoa.

• The Korean Journal of Physiology and Pharmacology
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• v.20 no.5
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• pp.547-556
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• 2016

Myometrial relaxation of mouse via expression of two-pore domain acid sensitive (TASK) channels was studied. In our previous report, we suggested that two-pore domain acid-sensing $K^+$ channels (TASK-2) might be one of the candidates for the regulation of uterine circular smooth muscles in mice. In this study, we tried to show the mechanisms of relaxation via TASK-2 channels in marine myometrium. Isometric contraction measurements and patch clamp technique were used to verify TASK conductance in murine myometrium. Western blot and immunehistochemical study under confocal microscopy were used to investigate molecular identity of TASK channel. In this study, we showed that TEA and 4-AP insensitive non-inactivating outward $K^+$ current (NIOK) may be responsible for the quiescence of murine pregnant longitudinal myometrium. The characteristics of NIOK coincided with two-pore domain acid-sensing $K^+$ channels (TASK-2). NIOK in the presence of $K^+$ channel blockers was inhibited further by TASK inhibitors such as quinidine, bupivacaine, lidocaine, and extracellular acidosis. Furthermore, oxytocin and estrogen inhibited NIOK in pregnant myometrium. When compared to non-pregnant myometrium, pregnant myometrium showed stronger inhibition of NIOK by quinidine and increased immunohistochemical expression of TASK-2. Finally, TASK-2 inhibitors induced strong myometrial contraction even in the presence of L-methionine, a known inhibitor of stretch-activated channels in the longitudinal myometrium of mouse. Activation of TASK-2 channels seems to play an essential role for relaxing uterus during pregnancy and it might be one of the alternatives for preventing preterm delivery.

• Korean Journal of Pharmacognosy
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• v.46 no.2
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• pp.167-172
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• 2015

Aleurone layer of Black rice (Oryza sativa L.) is enriched with anthocyanin that could increase bone density. This study was conducted to investigate the osteoporosis-preventing effects of the aleurone layer extract (BRE) on bone loss of ovariectomized (OVX) rats. OVX (or sham-operated) rats were assigned to three groups (n=8 per group): sham operated group (Sham); OVX control group (OVX); OVX-BRE group, OVX rats treated with BRE at 90 mg/kg B.W. The deionized water alone or deionized water with BRE was orally administrated to Sham, OVX or OVX-BRE groups, respectively for 12 weeks. High fat diet with 45 kcal% fat and water were fed to all rats ad libitum. Body weight was significantly decreased in the OVXBRE group compared to the OVX group (p<0.05). The bone mineral density and bone length of tibia were significantly higher in the OVX-BRE group compared to the OVX group and breaking force was significantly higher for the both tibia and femur bones. Serum estradiol concentration and calcium concentration of femur were higher in the OVX-BRE group than those of OVX group. However, serum alkaline phosphatase activity and parathyroid hormone concentration were decreased in the OVXBRE group compared to the OVX group. The results suggest that aleurone layer of Black rice is a potentially useful ingredient to protect against estrogen deficiency or menopause related osteoporosis.