• Asian-Australasian Journal of Animal Sciences
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• 제30권1호
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• pp.20-33
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• 2017

Objective: Superstimulatory treatment of one-month-old lambs can achieve synchronous development of numerous growing follicles. However, these growing follicles cannot complete maturation and ovulation. Oocyte maturation and competence are acquired during follicular development, in which granulosa cells play an essential role. Methods: In this study, we applied RNA sequencing to analyze and compare gene expression between prepubertal and adult superstimulated follicle granulosa cells in sheep. Results: There were more than 300 genes that significantly differed in expression. Among these differently expressed genes, many extracellular matrix genes (EGF containing Fibulin Like Extracellular Matrix Protein 1, pentraxin 3, adrenomedullin, and osteopontin) were significantly down-regulated in the superstimulated follicles. Ingenuity pathway and gene ontology analyses revealed that processes of axonal guidance, cell proliferation and DNA replication were expressed at higher levels in the prepubertal follicles. Epidermal growth factor, T-Box protein 2 and beta-estradiol upstream regulator were predicted to be active in prepubertal follicles. By comparison, tumor protein P53 and let-7 were most active in adult follicles. Conclusion: These results may contribute to a better understanding of the mechanisms governing the development of granulosa cells in the growing follicle in prepubertal sheep.

• Asian-Australasian Journal of Animal Sciences
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• 제20권6호
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• pp.842-849
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• 2007

The purpose of the present study was to produce live offspring from in vitro fertilized goat embryos. Oocytes were collected from abattoir ovaries and kept in oocyte collection medium. Oocytes were washed 4-5 times with maturation medium containing medium-199 with 5 ${\mu}g/ml$ FSH, 100 ${\mu}g/ml$ LH, 1 ${\mu}g/ml$ estradiol-$17{\beta}$ 50 ${\mu}g/ml$ gentamycin, 10% inactivated estrus goat serum, and 3% BSA (fatty acid free). Oocytes were placed in 100 ${\mu}l$ drops of maturation medium containing granulosa cell monolayer and incubated in a 5% $CO_2$ incubator at $38.5^{\circ}C$ for 27 h. For capacitation of spermatozoa fresh semen was processed and mixed in 3 ml fertilization TALP medium containing 50 ${\mu}g/ml$ heparin and kept in the above incubator for 2 h. The capacitated spermatozoa were coincubated with matured oocytes for fertilization. Cleaved embryos were separated and cultured in embryo development medium with oviductal cells and 494 embryos were produced. Recipient goats were synchronized with two injections of 15 mg $PGF_{{2}{\alpha}}$/goat 10 days apart. Eighty early stage embryos were transferred into the uterotubal junction of 14 surrogate mothers using laparoscopy techniques. One recipient delivered twin kids, whereas another two recipients each.delivered a single kid The parentage of these kids was evaluated using highly polymorphic co-dominant microsatellites markers. From the present study, it was concluded that live goat kids can be produced from in vitro matured and fertilized goat embryos, to the best of our knowledge for the first time in India.

• Nutrition Research and Practice
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• 제8권3호
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• pp.292-296
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• 2014

BACKGROUND/OBJECTIVES: This study was conducted to assess the potential of St. John's Wort (Hypericum perforatum) to prevent obesity and abnormalities in lipid metabolism induced by ovariectomy in a rat model without stimulatory activity on uterus. MATERIALS/METHODS: Ovariectomized (OVX) rats were treated for 6 weeks with 70% ethanol extracts of Hypericum perforatum [HPEs: whole plant (WHPE) and flower and leaves (FLHPE)], ${\beta}$-estradiol-3-benzoate at a dose of $50{\mu}g/kg/day$ (E2) or vehicle (distilled water). RESULTS: As expected, OVX increased body weight gain and adiposity and showed higher food efficacy ratio. OVX also increased the serum cholesterol as well as insulin resistance, while reducing uterus weight and uterine epithelial proliferation rate. HPEs (WHPE and FLHPE) showed estrogen-like effect on body weight gain, adipose tissue weight and food efficacy ratio in OVX rats. HPEs prevented hypercholesterolemia induced by OVX more effectively than E2. E2 increased uterus weight and epithelial proliferation rate in OVX rats, while HPEs maintained them at the level of the sham-operated animals. CONCLUSIONS: Our finding demonstrates that HPEs can be considered as an effective agent to prevent OVX-induced obesity without stimulatory activity on uterus.

• 대한수의학회지
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• 제43권4호
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• pp.649-656
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• 2003

We have a cultured method to grow rat mammary epithelial cells (RMEC) for 1 to 14 days in 1:1 mixture of Dulbecco's Modified Eagle Medium: Nutrient and F-12 (DMEM/F-12) containing 10% fetal bovine serum (FBS), human EGF, insulin, hydrocortisone, human transferrin and $17{\beta}$-estradiol in vitro. We were able to isolate and distinguish two cell types, luminal epithelial cells and myoepithelial cells, from primary clutures of RMEC. Immunocytochemical stains were used to distingusih luminal epithelial cells and myoepithelial cells. Peanut lectin (PNA) was stained in most alveolar epithelail cells and luminal epithelial cells of rats, while Thy-1.1, a maker of potential rat mammary myoepithelial cells, was expressed in myoepithelial cells in the rat. Also, we examined the expression patterns of three types of gap junction proteins, connexin 26 ($C{\times}26$), connexins 32 ($C{\times}32$) and connexin 43 ($C{\times}43$) by immunocytochemistry and western blot analysis. In the cell types, the results show that at the early stage of culture, luminal epithelial cells were increased and these cells were surrounded by myoepithelial cells. At the late stage of culture, luminal epithelial cells were decreased, in contrast myoepithelial cells were increased. In the expression pattern of gap junction, $C{\times}26$ maintained it's expression until day 3, but afterwards gradually decreased in intensity. Expression of $C{\times}32$ remained until day 5, then decreased slightly. $C{\times}43$ gradually increased untill the middle time of culture then decreased in intensity. These results suggest that connexins may be important for the control of growth in rat mammary epithelial cell types.

• Plant Resources
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• 제6권1호
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• pp.27-35
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• 2003

In this study, as preliminary research for the development of natural estrogen supplement the chemical properties of Iranian black pomegranate juice concentrate and seeds were evaluated. Proximate compositions of pomegranate juice concentrate and seeds were as follows; crude lipid 0.4% and 8.2%, moisture 39.9% and 6.6%, crude protein 0.9% and 12.2%, ash 1.4% and 1.7%, and carbohydrate 42.0% and 84.5% respectively. Major amino acids are glutamic acid (1310.0ppm) and aspartic acid (896.2ppm) in juice concentrate, and glycine (611.1ppm) and arginin (401.6ppm) in seeds. Ascorbic acid has the highest concentration of 20.0mg/l00g in juice concentrate and 0.23mg/l00 in seeds. The compositions of unsaturated fatty acids such as linoleic acid and linolenic acid were higher than those of saturated fatty acids such as stearic palmitic acid. Major minerals were potassium, calcium and sodium, potassium was highest in both juice concentrate and seeds. Vitamins were composed of ascorbic acid (20.0mg/l00g), vitamin B$_1$(0.12mg/100g) and niacin (0.80mg/l00g) in juice concentrate, and only ascorbic acid(0.23mg/l00g) in seeds. Organic acids such as citric and L-malic acid were detected only in pomegranate juice concentrate. The contents of total polyphenols were 4.55g/L in juice concentrate and 3.5mg/l00g in seeds, respectively. Phytoestrogens detected in pomegranate juice concentrate and seeds were daidzein, quercetin, genistein and 17 $\beta$-estradiol.

• 혜화의학회지
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• 제20권1호
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• pp.91-104
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• 2011

Background : Although chronic non-bacterial prostatitis is increasing, it is hard to treat effectively. In western medicine, antimicrobials drug, ${\alpha}$-adreno-ceptor antagonists, anti-inflammatory drugs, tricyclic antidepressants and anticholinergic agents are used commonly, but chronic prostatitis/chronic pelvic pain syndromes is confusing and frustrating for urologist. IDS(Indongsoyeom-bang) is used in treatment of chronic prostatitis/chronic pelvic pain syndromes. And it is reported that GLS(Gleditsiae spina) and TOF(Toosendan fructus) components of IDS have significant effect on protection of the glandular epithelial cells. Objective : In this study was conducted to investigate the therapeutic effects and action machanism of IDS in the rat model of non-bacterial prostatitis induced by castration and testosterone treatment. Methods : We observed six experimental objects of normal group, control group, testosterone group, and IDS 50 mg/kg, 200mg/kg, 400mg/kg group. Rats were treated with 17 ${\beta}$-estradiol after castration for induction of experimental non-bacterial prostatitis, which is similar to human chronic prostatitis in histophatological profiles. IDS and testosterone were administered as an experimental specimen and a positive control, respectively. The prostates were evaluated by histological parameters including the epithelial score and epithelio-stromal ratio for glandular damage. Also, the prostates were observed by Hematological alterations of WBC, RBC, hemoglobin and platelet. Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with IDS-50 showed a diminished range of the tissue damage. Epithelial score was improved in IDS than that of the control. The epithelio-stromal ratio was lower in IDS when compared to that of the control. Also, the examination of bloods were not observed hematological change. Conclusion : These finding suggests that IDS may protects the glandular epithelial cells. We concluded that IDS could be a useful remedy agent for treating chronic non-bacterial prostatitis.

• 대한의생명과학회지
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• 제20권3호
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• pp.156-161
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• 2014

It has been reported that safflower seeds have preventive effects against osteoporosis. Recently, safflower buds (SB) were found to have more useful functional ingredients than safflower seeds. In the current study, we evaluated the anti-osteoporosis effects of SB diet in ovariectomized (OVX) rats. The rats were divided into five groups; sham operated group, OVX alone group, OVX plus $17{\beta}$-estradiol ($E^2$ $10{\mu}g/kg$, i.p.) and OVX plus SB diet feeding group (0.3% or 1%). Feeding of SB diet (0.3% or 1%) to OVX rats markedly increased bone mineral density (BMD) of femurs, compared to the OVX group. The OVX rats exhibited a marked increase in trabecular separation (Tb.Sp) and this change was inhibited by the feeding of SB diet, similar to that seen with OVX+E2 group. Moreover, feeding of SB diet to OVX rats decreased the markers of bone turnover, including osteocalcin and alkaline phosphatase (ALP). These results suggest that SB extract has a bone sparing effect in OVX-induced trabecular bone loss and prevents deterioration of bone microarchitecture by suppressing the rate of bone turnover. Therefore, SB may be useful for preserving bone mass and structure in estrogen deficient women with a potential role in reducing postmenopausal osteoporosis.

• Journal of Preventive Medicine and Public Health
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• 제43권4호
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• pp.301-308
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• 2010

Objectives: In most DEHP exposure assessment studies, single spot urine sample was used. It could not compare the exposure level among studies. Therefore, we are going to represent the necessity of selection of proper sampling time of spot urine for assessing the environmental DEHP exposure, and the association urinary DEHP metabolites with steroid hormones. Methods: We collected urine and plasma from 25 men. The urine sampling times were at the end of the shift (post-shift) and the next morning before the beginning of the shift (pre-shift). Three metabolites of DEHP {mono(2-ethylhexyl) phthalate [MEHP], mono-(2-ethyl-5-hydroxyhexyl)phthalate [MEHHP], and mono(2-ethyl-5-oxohexyl)phthalate [MEOHP]} in urine were analyzed by HPLC/MS/MS. Plasma luteinzing hormone, follicle stimulating hormone, testosterone, and $17{\beta}$- estradiol were measured at pre-shift using a ELISA kit. A log-transformed creatinine-adjusted urinary MEHP, MEHHP, and MEOHP concentration were compared between the post- and pre-shift. The Pearson’s correlation was calculated to assess the relationships between log-transformed urinary MEHP concentrations in pre-shift urine and hormone levels. Results: The three urinary metabolite concentrations at post-shift were significantly higher than the concentrations in the pre-shift (p<0.0001). The plasma hormones were not significantly correlated with log-transformed creatinine - adjusted DEHP metabolites. Conclusions: To assess the environmental DEHP exposure, it is necessary to select the urine sampling time according to the study object. There were no correlation between the concentration of urinary DEHP metabolites and serum hormone levels.

• Asian Pacific Journal of Cancer Prevention
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• 제16권17호
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• pp.7575-7582
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• 2015

Cyclin E, a key coordinator of the G1 to S transition in the cell cycle, may be deregulated in several malignancies, including breast cancer. The most significant aberration in cyclin E is its elastase mediated proteolytic cleavage into tumor specific low molecular weight isoforms (LMW-Es). LMW-Es are biochemically hyperactive and biologically drive tumorigenesis in transgenic mouse models. Additionally, expression of LMW-Es has been correlated with poor survival in breast cancer cases. Here we determine whether expression of LMW-Es in a breast cancer cell line that is naturally devoid of these deregulated forms would alter their progression through each phase of the cell cycle. The results revealed that LMW-Es expression resulted in an increased doubling time, concomitant with a predominant increase in the population in the S phase of the cell cycle. Moreover, downregulation of p53 in LMW-Es cells resulted in additional shortening of the doubling time and enrichment of cells in the S and G2/M phases of the cell cycle. Furthermore, expression of LMW-Es sensitized cells to ${\beta}$-estradiol (E2) mediated growth and changed expression patterns of estrogen receptor and Bcl-2. Intriguingly, expression of LMW-Es could surpass anti-apoptotic effects raised by p53 upregulation. Taken together these studies suggest that overexpression of LMW-Es in collaboration with p53 loss results in altered g rowth properties of MCF-7 cells, enhancing the oncogenic activity of these ER positive breast cancer cells.

• Archives of Pharmacal Research
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• 제26권11호
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• pp.917-924
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• 2003

This research aims to test a new drug candidate based on a traditional medicinal herb, F1, an herbal extract obtained from Astragalus membranaceus and its main ingredient, 1-monolinolein that may have fewer side effects and less uterine hypertrophy. In vitro experiments, human osteoblast-like cell lines, MG-63 and Saos-2, were analyzed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and an alkaline phosphatase (ALP) assays. Mouse osteoclasts were induced through a calcium-deficient diet and inhibition effects were measured. In vivo experiments were done using ovariectomized (OVX) rats for 9 weeks. At necropsy, uterus weights were measured, trabecular bone area (TBA) of tibia and lumbar vertebra were measured bone histomorphology. In results, cell proliferation and ALP activity in Saos-2 by ether F1 or 1-monolinolein did not increased significantly compared to the control. The F1 inhibited osteoclast development ($IC_{25}=3.37{\times}10^{-5}$mg/mL) less than 17$\beta$-estradiol. The OVX rats administered F1 (2 mg/kg/day and 10 mg/kg/day) showed an increase in TBA of the tibia significantly (136.3${\pm}4.2% and 138.5{\pm}$10.3% of control). In conclusions, the herbal extract, F1 inhibited tibia and lumbar bone loss and did not cause uterine hypertrophy. However, 1-monolinolein, the main ingredient of the herbal extract, did not inhibit bone loss.