• 대한수의학회지
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• 제28권1호
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• pp.203-211
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• 1988

The present study was carried out to investigate the estrous behavior and vaginal smear after induction of estrus with exogenous hormones in the premature, metestrous and anestrous bitches. A total of 21 bitches(Mixed breed: 16, Jindo breed: 5), from 10 months to 5 years of age and weighing 8 to 15 kg was studied the change of vaginal smear and the estrous behavior before and after induction of estrus. The results obtained are as follows: 1. In the treatment A(They were given the $PGF_2{\alpha}$, estrone, estradiol-$17{\beta}$, PMSG and HCG) proestrus commenced in $10.16{\pm}1.44$($Mean{\pm}SEM$) days after treatment, The mean duration of proestrus, and estrus was $7.50{\pm}1.44$ and $13.50{\pm}3.44$ days, respectively. In the treatment B(They were given the PMSG and HCG) proestrus commenced in $5.53{\pm}0.59$ days after treatment. The mean duration of proestrus and estrus was $6.60{\pm}0.71$ and $14.60{\pm}1.14$ days, respectively. 2. All of the 6 bitches in the treatment A showed vulval swelling and vaginal discharge, 14 of the 15 bitches in the treatment B showed vulval swelling and vaginal discharge. However, all of the treatment A and B showed male acceptance. 3. The main change of vaginal smear in proestrus and estrus after induction of estrus was a increase in the proportion of anuclear and superficial cells associated with a decrease in small intermediate and parabasal cells. By the estrous behavior and vaginal smear the estrus was induced in all the premature, metestrous and anestrous bitches.

• 대한본초학회지
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• 제22권4호
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• pp.145-153
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• 2007

Objective : Although chronic non-bacterial prostatitis is a common disease, it is very difficult to treat effectively. Toosendan fructus(TOF) has been traditionally used in treatment of abdominal pain and parasite. In this study, we investigated the therapeutic effects and action mechanism of Toosendan fructus (TOF) in the rat model of non-bacterial prostatitis induced by castration and testosterone treatment. Method : Eight-month-old 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. Toosendan fructus(TOF) and testosterone were administered as an experimental specimen and a positive control, respectively. The prostates were evaluated by histopahological parameters including the epithelial score and epithelio-stromal ratio for glandular damage. Also, the prostates were observed by hematological alterations of WBC, RBC, hemoglobin, hematocrit and platelet. Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with Toosendan fructus(TOF) showed a diminished range of the tissue damage. Epithelial score was improved in Toosendan fructus(TOF) than that of the control. The epithelio-stromal ratio was lower in Toosendan fructus(TOF) when compared to that of the control. Also, the examination of bloods were not observed hematological change. Conclusions : These findings suggest that Toosendan fructus(TOF) may protect the glandular epithelial cells and may take hematological safety. We concluded that Toosendan fructus(TOF) may could be a useful remedy agents for treating the chronic non-bacterial prostatitis.

• 한국가축번식학회지
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• 제10권2호
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• pp.211-217
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• 1986

This experiment was conducted to determine the effect of FSH on in vitro maturation and in vitro fertilizing ability of oocytes recovered from normal follicles of different sizes in superovulated rabbits. Follicular oocytes recovered were cultured in modified Ham's F12 medium containing 0, 0.1, 1.0 and 10$\mu\textrm{g}$ FSH/ml for 18 hours and investigated the degree of cumulus cells expansion and nuclear maturation, which were fertilized with in vivo capacitated rabbit sperm. 1. The number of normal follicles<1.5mm, 1.6 to 2.5mm and> 2.5mm in diameter at 16 to 18hrs after HCG administration was 4.8 (38.8%), 5.5(45.4%) and 3.3(15.8%), respectively. Average percent of oocytes recovered was 69.7% and larger follicles tended to have a higher percent, recovery rate than smaller follicles. 2. The degree of cumulus expansion in medium containing 0.1$\mu\textrm{g}$ FSH/ml was similar to that of control, but markedly decreased under the level of above 1$\mu\textrm{g}$ FSH/ml. The proportions of oocytes which reached the second meiotic metaphase were 57.1, 61.5, 43.8 and 45.0% in medium containing 0, 0.1, 1.0 and 10$\mu\textrm{g}$ FSH/ml, respectively. Oocytes from larger follicles showed a higher nuclear maturation than that from smaller follicles. 3. In vitro fertilization rate of oocytes matured under 1$\mu\textrm{g}$ FSH/ml was slightly, not significant, higher than that of others. 4. Progesterone level in follicular fluid was about 67 to 71ng/ml with no difference in follicular sizes and estradiol-17$\beta$ level was under 25pg/ml.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.71-71
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• 2003

Human umbilical cord blood cells(HUCBC) are rich in mesenchymal progenitor cells, endothelial cell precursors and hematopoietic cells. HUCBC have been used as a source of transplantable stem and progenitor cells. However, little is known about survival and development of HUCBC transplantation in the CNS. Estrogen has a neuroprotective potential against oxidative stress-induced cell death so has an effect on reducing infarct size of ischemic brain. We investigated the potential use of HUCBC as donor cells and tested whether estrogen mediates intravenously infused HUCBC enter and survive in ischemic brain. PKH26 labeled mononuclear fraction of HUCBC were injected into the tail vein of ischemic OVX rat brain with or without $17\beta$-estradiol valerate(EV). Under fluorescence microscopy, labeled cells were observed in the brain section. Significantly more cells were found in the ischemic brain than in the non-ischemic brain. HUCBC transplanted into ischemic brain could migrate and survive. Some of cells have shown neuronal like cells in hippocampus, striatum and cortex tissues. These result suggest that estrogen reduces ischemic damage and increases the migration of human umbilical cord blood cells. This Study was supported by the Korea Science and Engineering Foundation(KOSEF) though the Biohealth Products Research Center(BPRC), Inje University, Korea.

• Molecular & Cellular Toxicology
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• 제1권3호
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• pp.149-156
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• 2005

Di (n-ethylhexyl) phthalate (DEHP) is thought to mimic estrogens in their action, and are called endocrine disrupting chemicals. DEHP is used in numerous consumer products, especially those made of flexible polyvinyl chloride and have been reported to be weakly estrogenic. In this study, DEHP were tested for estrogenic properties in vitro models and with microarray analysis. First, the E-screen assay was used to measure the proliferation of DEHP in MCF-7 cells, a human breast cancer cell line. DEHP induced an increase in MCF-7 cell proliferation at concentration of $10^{-4}M$. Second, we carried out a microarray analysis of MCF-7 cells treated with DEHP using human c-DNA microarray including 401 endocrine system related genes. Of the genes analyzed, 60 genes were identified showing significant changes in gene expression resulting from DEHP. Especially, 4 genes were repressed and 4 genes were induced by DEHP compared to $17{\beta}-estradiol$. Among these genes, trefoil factor 3 (intestinal), breast cancer 1, early onset and CYP1B1 are involved in estrogen metabolism and regulation. Therefore it suggests that these genes may be associated with estrogenic effect of the DEHP on transcriptional level. The rationale is that, as gene expression is a sensitive endpoint, alterations of these genes may act as useful biomarkers to define more precisely the nature and level of exposure to kinds of phthalates.

• 한국가축번식학회지
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• 제21권2호
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• pp.95-102
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• 1997

The studies on the carried out to investigate the effective concentration of cryoprotectant agents and sucrose by one-step straw method of bovine embryos. The follicular oocytes were cultured in TCM-199 medium containing 10 IU/ml PMSG(Sigma, USA), 10 IU/ml hCG(Sigma, USA), 1$\mu\textrm{g}$/ml $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The bovine embryos following dehydration by cryoprotective agents and various concentrations of sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water. Survival and in vitro developmental rate was defined as devellpmental rate on in vitro culture or FDA-test. The results are smmarized as followes : 1. The high in vitro developmental rates of bovine frozen embryos after rapidly thawed in freezing medium was attained 2.0M glycerol, 2.0M DMSO, 1M or 2.0M propanediol. 2. The high in vitro developmental rates of bovine frozen embryos after rapidly thawed in freezing medium was obtained single cryoprotectant(6.7~17.4%) than mixed cryoprotectants(6.7~16.7%). 3. In vitro developmental rate of bovine embryos after rapid frozen-thawing in the freezing medium added 0.25M and 0.50M sucrose were higher cleavage rate than those of sucrose concentration of 0.75M and 1.00M. 4. The freezing methods on in vitro developemental rates of bovine embryos was attained slow freezing method(9.70~15.6%) higher than rapid freezing method(9.4~13.3%).

• 한국가축번식학회지
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• 제21권2호
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• pp.111-116
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• 1997

We determined the effects of follicular fluid fractions in the maturation medium on bovine oocyte maturation, fertilization and subsequent development, as well as on number of cells in blastocysts following culture. Follicular fluid and oocytes from bovine follicles less than 5 mm in diameter were collected from the ovaries of slaughtered cows. Follicular fluid was separated into different molecular weight fractions by untrafiltration through a membrane using a centrifuge at 500$\times$g, for 2h. For the maturation medium, follicular fluid fractions (30%, v/v), whole fluid (30%) or PVP(3mg/ml) were added to TCM 199(0.1$\mu\textrm{g}$/ml estradiol-17$\beta$, 100IU hCG). After maturation for 24h, oocytes were fertilized in vitro with bull frozen-thawed spermatozoa and cultured on a monolayer of granulosa cells for 9 days after fertilization. There were no differences in maturation rates or fertilization rates among any maturation conditions. The rates of development to >2-cell stage of the oocytes were significantly decreased when fraction of follicular fluid below 10,000 MW were added into maturation medium, compared with control and fraction above 10,000 MW(26.0% vs 40.8% to 64.0%, respectveily. p<0.01). Likewise, the rates of development to blastocysts of fertilized oocytes were significantly decreased in maturation medium containing fraction of follicular fluid (<10,000 MW). The average cell number of blastocysts derived from oocytes that matured in the fraction(>10,000 MW) of follicular fluid was 154.7$\pm$13.7. These embryos contained more cells than those matured in whole follicular fluid, or the fraction(<10, 000 MW) of follicular fluid or control(107.0$\pm$8.4, 91.8$\pm$11.8 and 95.8$\pm$6.2, respectively). In conclusion, we found that fractions of follicular fluid contained factors stimulating or inhibiting oocyte cytoplasmic matruation. These suggest that a factor(s) inducing cytoplasmic maturation of oocytes may exist in >10,000 MW fraction of follicular fluid.

• 한국수정란이식학회지
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• 제7권2호
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• pp.125-131
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• 1992

The nudear changes of bovine oocytes during 24 hrs. of culture for mejotic maturation were examined. Bovine oocytes were collected from small(<2 mm), medium(2~6 mm) and large(>6mm) follicles and classified into three grades by their morphological characteristics. A total of 242 oocytes collected were obtained:from 184 small, 157 medium and 1 large follicles, respectively and were classified into 95 grade I, 155 grade H and 92 grade III oocytes. All the bovine oocytes collected and graded were washed with a basal medium and incubated in groups of 10 for 24 hrs in 5% $CO_2$ and 39$^{\circ}C$. The basal medium used was composed of TCM-199 supplemented with sodium bicarbonate, sodium pyruvate, streptomycin, penicillin G and 10% FCS. The oocytes were cultured in drops of 50,$\mu$l basal medium supplemented with 35$\mu$g /ml FSH, 10$\mu$g /ml LH and 1$\mu$g /ml estradiol-17$\beta$. The oocytes were fixed and examined on their chromosomal status by 1% acetorcein staining in the interval of 3 hrs. Most of the grade I oocytes developed to germinal vesicule stage at 0 to 3 hrs., germinal vesicle breakdown at 6 hrs., metaphase I at 9 to 15 hrs., anaphase I and telophase I at 18 hrs., and metaphase II and the first polar body at 24 hrs. after culture for meiotic maturation. However, it was found that compared to grade I oocytes, grade H and W oocytes reached earlier to germinal vesicle breakdown and most of them developed earlier to M II stage at 21 hrs. after culture.

• 한국수정란이식학회지
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• 제23권4호
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• pp.229-237
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• 2008

Recently, parabens have been believed to act as xenoestrogens, an identified class of endocrine disruptors (EDs). These environmental compounds are the most well-known as preservatives in many commercial products, including food, cosmetics and pharmaceutical industries. It has been demonstrated that the human health risks of parabens result from a long-term exposure to skin in which this chemical group is rapidly absorbed through the skin. On the other hand, parabens are also completely absorbed from gastrointestinal tract. It has reported that these substances possess several biological effects in which inhibitory property involved in membrane transports and mitochondrial functions is considered to be important for their action. Testing of parabens has revealed that estrogen-like activities of these chemicals are much less potent than natural estrogen, $17{\beta}$ estradiol (E2). Additionally, the estrogenicity of individual paraben- compounds is distinct depending upon their biochemical structure. Recent findings of paraben-estrogenic activities have shown that these compounds may affect breast cancer incidence in women, suggesting adverse ecological outcomes of this environmental group on human and animal health. Although the biological and toxicological effects of parabens have been demonstrated in many previous studies, possible mechanism(s) of their action are required to be explored in order to bring the better understanding in the detrimental impacts of parabens in human and wildlife. There have several different types of parabens which are the most widely used as preservatives. These include methyl-paraben, ethylparaben, propylparaben, butylparaben and p-hydroxybenzoic acid, a major metabolite of parabens. In this review, we summarize current database based on in vitro and in vivo assays for estrogenic activities and health risk assessment of paraben- EDs which have been published previously.

• 대한수의학회지
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• 제29권3호
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• pp.253-262
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• 1989

This study was performed to investigate the patterns of progesterone secretion after induction of estrus in premature, metestrous and anestrous bitches. A total of 22 bitches were used. Of them 18 bitches were treated with hormone to induced estrus and 4 bitches were untreated and served as controls. Estrus was induced with $PGF_{2{\alpha}}$, estrone, estradiol-$17{\beta}$, PMSG and HCG(Treatment A), and with PMSG and HCG(Treatment B). Blood samples were collected via the cephalic vein at 2 to 5 days interval. Blood samples were centrifuged (1,200g, 10min.) within 30 minutes after collection and plasma was stored at $-20^{\circ}C$ until analyzed for the progesterone concentrations. Plamsa progesterone concentrations were measured by radioimmunoassay. The results of estrous induction were determined by estrous signs, ovarian response, egg recovery and progesterone patterns. The results obtained were as follows; 1. All bitches in treatment A showed estrous signs, however the ovarian response and egg recovery were not detectable and the levels of progesterone were nearly same as before. 2. In the treatment B, premature and metestrous bitches showed only estrous signs, however 5 of 7 anestrous bitches (71.4%) showed estrous signs, ovarian response and changes of progesterone levels. In conclusion, clinical estrous behavior can be induced during any phase of the estrous cycle, but ovulation should be induced only if induction occur approximately 4 months or more after the previous estrus.