• International journal of advanced smart convergence
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• v.8 no.1
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• pp.184-195
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• 2019

This study investigates the hair restoration efficacy of selected radish saponin extracts on nude mice. Nude mice genetically predisposed to pattern balding were used in this study. Our study revealed the underlying mechanism of stimulating hair growth in athymic nude mice by repair the nu/nu follicular keratin differentiation defect. Thus, the topical application of radish saponin may represent a novel strategy for the management and therapy of certain forms of alopecia. The term of hair density of PEE treated nude mice were significantly increase as compared with of control nude mice. Histological observation of skin sample showed no hair follicle or only distorted hair follicles were observed in the control samples, in contrast, by the PEE treatment groups showed a fully formed and increased the number of hair follicles up to three times higher than that of control group in terms of the number of hair follicles in nude mouse skin.PEE treated mice the number of BrdU-labeled keratinocytes per anagen follicle increased significantly, especially in the follicular bulbs and outer root sheath compared with the control mice. Moreover, PEE-treated nude mice also exhibited a significant increase in the number of BrdU-labeled epidermal keratinocyte proliferation.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.95-95
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• 2003

Endocrine factors, such as steroid hormones and growth factors, regulate egg productivity including the number of egg production, egg weight, sexual maturity, and the number of small yellow follicles. Especially, insulin-like growth factor-I (IGF-I) is involved in the regulation of ovulation rate and ovarian follicular development in chickens, and the relationship between IGF-I genotype and egg weight was reported. However, the effect of grwoth factors on egg productivity in Korean Native Ogol Chicken (KNOC) has not been studied. Therefore this study was conducted to identify the relationship among endocrine factors, IGF-I genotypes, and egg productivity. IGF-I genotypes (AA, AB, BB) were represented to 12.6%, 34%, and 53.4%, respectively. AB genotype stimulates the secretion of estradiol and progesterone in serum (30 and 40 week), regulates growth and proliferation of follicles at 60 weeks, and is positively associated with the number of small yellow follicles. Therefore, these results suggest that there are possibility to IGF-I genotypes for a genetic marker in egg productivity of KNOC.

• Journal of Embryo Transfer
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• v.10 no.1
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• pp.45-51
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• 1995

In order to determine the optimum condition and timing for in vitro maturation of oocytes to metaphase of meiosis II (M II), the immatured follicular oocytes were recovered by puncturing the large(1.0~1.5 mm in diameter) and small(<1.0 mm in diameter) follicles in the ovaries of rabbits treated intramuscularly with a single dose of 100 TU PMSG 68 hours previously. The follicular oocytes were classified into three grades by the attachment of cumulus cells. The Grade I and II follicular oocytes from large follicles were cultured in BO-DM medium with 10% FCS, 35 $\mu$g /nl of FSH, 10 $\mu$g /ml of LH and 1 $\mu$g /ml of estradiol-17$\beta$ at 39t in a 5% $CO_2$ incubator for 11 to 23 hours. In 3 hours interval during the culture period, the oocytes were harvested and their cumulus cells were removed with hyaluronidase. The denuded oocytes were stained with Hoechst 33342 dye and their meiotic status and extrusion of the first polar body (PB) were examined under a fluorescence microscope. Also the fragmentation of the first PB and the distance between the first PB and nucleus were examined. The results obtained were as follows: 1. The mean recovery rate of follicular oocytes from the large and small follicles was 59. 9 and 31.3%, respectively. The mean number of oocytes recovered per rabbit and the Grade I percentage were 14.6 and 94.4% in large follicles, but 2.1 and 61.1% in small follicles, respectively. All the parameters examined were different significantly (p<0.05) between both the folliclular size. 2. Most of the follicular oocytes(86.8%) were matured in vitro to M II phase in 14 hours in Grade I oocytes, but the significantly(p<0.05) less oocytes(45.5%) were matured in Grade II oocytes. 3. The first PB was extruded in most of the oocytes(94.7%) in 14 hours of culture with the fragmentation rate of 29.6%, but the fragmentation rate of the first PB increased significantly (p<0.05) as the culture period for maturation was longer to 20 hours(63.5%). 4. The distance between the first PB and nucleus was increased linearly (p<0.05) as the maturation time passed from 14(7.1$\mu$rn) to 23 hours(58.4$\mu$m). 5. From the above results it was concluded that the optimum time for in vitro maturation culture might be 14 hours in the follicular oocytes from rabbit primed with PMSG for 68 hours, expecially when these follicular oocytes were used for recipient cytoplasms in embryo cloning.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.9
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• pp.1205-1209
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• 2000

The present study was undertaken to investigate the effect of stimulation of follicular development with eCG on the peripheral levels of inhibin and FSH in Murrah buffaloes. Estrus was synchronized in five normally cycling females by insertion of Crestar (Intervet, Boxmeer, Holland) implants for nine days. Estradiol valerate was administered i.m. on the day of implant insertion. On the 10th day of the induced estrous cycle a single dose of 3000 IU eCG (Folligon, Intervet, Boxmeer, Holland) was given, followed by treatment with 25 mg of $PGF_2$ alpha (Lutalyse, Upjohn, Belgium) 48 h later. Blood samples were obtained during the induced estrus, on cycle day 10 (luteal phase), at the superovulatory estrus (43 h after PGF) and during the periovulatory period (64 h after PGF). Ultrasonography was done daily to monitor follicular development. Plasma concentrations of inhibin and FSH were determined by specific radioimmunoassays. Differences between $mean{\pm}SEM$ values of different phases of the cycle were compared by ANOVA. The mean number of small (2-5 mm), medium (6-9 mm) and large (>10 mm) follicles observed two days after eCG treatment and on the day of superovulatory estrus was $2.8{\pm}0.31$, $5.2{\pm}0.30$ and $1.4{\pm}0.09$ and $1.9{\pm}0.21$, $2.8{\pm}0.40$ and $5.0{\pm}0.83$, respectively. The mean number of ovulations was $3.6{\pm}0.37$ and the mean number of unovulated follicles was $6.1{\pm}0.47$. Most of the follicles >10 mm in diameter had ovulated (72%). The mean ${\pm}SEM $ of plasma inhibin concentration $(2584.15{\pm}17.92pg/ml)$ during the superovulatory estrus was significantly higher $(p{\leq}0.05)$ than during the induced estrus $(749.87{\pm}17.29pg/ml)$, the luteal phase $(1099.54{\pm}24.98pg/ml)$ and periovulatory period $(1682.71{\pm}29.88pg/ml)$, respectively. $Mean{\pm}SEM$ plasma FSH concentration during the induced estrus $(10.35{\pm}0.41ng/ml)$ was not different from that during the superovulatory estrus $(8.52{\pm}0.39ng/ml)$, but was significantly higher $(p{\leq}0.05)$ than during the luteal phase $(2.81{\pm}0.42ng/ml)$ and periovulatory period $(5.7{\pm}0.28ng/ml)$. These data indicate that treatment with eCG in buffaloes for inducing superovulation results in a significant elevation in plasma inhibin levels and a decrease in plasma FSH levels during the superovulatory estrus. Thus, we suggest that the elevated plasma inhibin coming from fully developed follicles continued for a long time which results in inhibition of FSH leading to poor ovulation in the remaining follicles, which may be the cause of suboptimal superovulatory response.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.101-111
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• 2001

The present study was conducted to develop an in vitro culture system that would support bovine follicle growth from preantral to antral stage, oocyte maturation, fertilization, and embryonic development. Bovine preantral follicles (150$\pm$1.2 ${\mu}{\textrm}{m}$) surrounded by theca cell were isolated ezymatically and mechanically from ovarian cortical slides in Leibovitz L-l5 medium containing 1 mg/$m\ell$ collagenase and 0.2 mg/$m\ell$ DNase I and cultured for 25 days in the presence of different concentrations of bovine FSH and LH in $\alpha$MEM medium. The survival and growth rates of follicles cultured in the presence of FSH (10~150 ng/$m\ell$) were significantly higher than those of control group (P ＜ 0.001), but no significant differences were observed in survival and growth rates of follicles between the LH treatment groups (1~125 ng/$m\ell$) and the control. The survival (40%) and growth (244 $\pm$ 0.5 $\mu\textrm{g}$) of follicles cultured with FSH (90 ng/$m\ell$) and LH (25 ng/$m\ell$) were higher than those of control (25%, 160 $\pm$1.0 $\mu\textrm{g}$). Finally, 50% percent of healthy antral follicles were obtained, and almost 60% of them has complete meiotic division with 1st polar body (18.1%) and 10.0% have developed to the cleaved embryo and blastocyst stage. These results suggest that bovine preantral follicle with intact theca cell can grow to the antral stage using these culture conditions, and that oocytes from in vitro-matured bovine preantral follicle may acquire meiotic competence and can undergo fertilization and development.

• The Journal of Korean Medicine
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• v.31 no.2
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• pp.48-63
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• 2010

Objectives: Yonnyuniksoogobon-dan (Yan Nian Yi Shou Gu Ben Dan 延年益壽固本丹) is composed of 11 herbs (Polygoni Mutiflori Radix, Lycii Radicis Cortex, Polia, Rehmanniae Radix, Rehmanniae Radix Preparat, Asparagi Radix, Liriopis Tuber, Lycii Fructus, Acori Graminei Rhizoma, Angelicae Acutiloba Radix, and Pini Folium) based on Yonryunggobon-dan (Yan Ling Gu Ben Dan 延齡固本丹) and Yonnyuniksoobulrho-dan (Yan Nian Yi Shou Bu Lao Dan 延年益壽不老丹). This study evaluated hair growth promoting effect of Yonnyuniksoogobon-dan on the shaved C57BL/6 mice. Methods: Yonnyuniksoogobon-dan was treated by oral administration (Sample I) and oral administration plus skin application (Sample II) once a day for 12 days. Hair regrowth was photographically and histologically determined during the experimental period. Hair growth cycle related factors (EGF, TGF-${\beta}1$) and vascular factors (VEGF, iNOS) were also determined with immunohistochemistry. Results: 1. On gross observation of hair regrowth, Sample I and Sample II groups demonstrated acceleration of hair regrowth. 2. The hair regrowth index of the Sample I group increased significantly from 7 days (P<0.05) to 12 days (P<0.01) after the shave while that of the Sample II group significantly increased at 12 days (P<0.05). 3. On histological observation, both Sample I and Sample II groups demonstrated histological improvement and increases of number and diameter of the hair follicles. 4. EGF expressions on the root sheath of hair follicles were up-regulated in both Sample I and Sample II groups. 5. TGF-${\beta}1$ expressions on the root sheath of hair follicles were not regulated in Sample I or Sample II groups. 6. VEGF expressions in the surrounding tissues of hair follicles were up-regulated in both Sample I and Sample II groups. 7. iNOS expressions in the surrounding tissues of hair follicles were down-regulated in both Sample I and Sample II groups. Conclusions: These results suggest that Yonnyuniksoogobon-dan has hair growth-promoting activity and these effects relate to up-regulations of EGF and VEFG expressions and down-regulations of TGF-${\beta}1$ and iNOS expressions on hair roots.

• The Journal of Korean Medicine
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• v.29 no.5
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• pp.77-89
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• 2008

Objectives : This study evaluates the hair growth promoting effect of Yonnyuniksoogobon-dan on shaved C57BL6 mice. Methods : Yonnyuniksoogobon-dan was administered orally (Group I) and both orally and by skin application (Group II) once a day for 12 days. The experimental groups were compared to Control, which was orally administered physiological saline solution. Hair regrowth was photographically and histologically determined during the experimental period. The levels of hair growth cycle related factors (EGF, TGF-${\beta}$1) and vascular factors (VEGF, iNOS) were also determined by immunohistochemistry. On gross observation of hair growth, both Group I and Group II shaved C57BL6 mice showed accelerated hair regrowth. Results : The hair regrowth index of Group I increased significantly from 7 days (P<0.05) to 12 days (P<0.01) after shaving and that of Group II was significantly higher at 12 days (P<0.05). On histological observation, both Group I and Group II demonstrated histological improvement and increases in the number and diameter of the hair follicles. EGF expression on the root sheath of hair follicles was up-regulated in both Group I and II. TGF-${\beta}$1 expression on the root sheath of hair follicles was unchanged in both Group I and II. VEGF expression in the tissues surrounding hair follicles was up-regulated in both groups. iNOS expression in the tissues surrounding hair follicles was down-regulated in both groups. Conclusions : These results suggest that Yonnyuniksoogobon-dan promotes hair growth and this effect is related to up-regulation of EGF and VEFG expression and down-regulation of TGF-${\beta}$1 and iNOS expression on hair roots.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.6
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• pp.638-642
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• 2001

To investigate the production of $C_{21}$-steroids during the spawning period of spotted sea bass, Lateolabrax maculatus, we have incubated maturing and ovulating follicles with radiolabeled pregnenolone and $17\alpha$-hydroxyprogesterone for 24 hours. The resulting metabolites were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), When maturing follicles ($700\sim800{\mu}m$ in diameters) were incubated with radiolabeled precursors, $C_{21}$-metabolites were corticosteroids and $17\alpha$-hydroxy, $20\beta$-dihydroprogesterone ($17\alpha20\beta OHP$). When ovulation follicles ($1,000\sim1,150{\mu}m$ in diameters) were incubated with radiolabeled precursors, the major $C_{21}$-metabolites were $17\alpha20\beta OHP$, $17\alpha$,$20\beta$, 21-trihydroxy-4-pregnen-3-one ($17\alpha20\beta21P$), and corticosterone. Additional chromatography by TLC and HPLC confirmed the presence of radioactive $17\alpha20\beta OHP$ in the maturing follicles, and $17\alpha20\beta OHP$,$17\alpha20\beta21P$ and corticosterone in ovulating follicles. Although $17\alpha20\beta OHP$ was found in a small peak, the synthesis of this steroid suggests that it may play a role in regulating the oocyte maturation process. Whereas ovulation is regulated by both $17\alpha20\beta OHP$ and $17\alpha20\beta21P$ in the spotted sea bass. In addition, an unusual finding was the biosynthesis of corticosterone. Whether this production is responsible for the ovulation, and is an area requiring continued research.

• The Korean Journal of Zoology
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• v.31 no.3
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• pp.177-184
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• 1988

The pattern of progesterone production and secretion of frog(R. dybowskii) follicles was investigated in follicle culture in vitro. Involvement of cAMP in the regulation of the steroid production by the follicles was also investigated by manipulating endogeneous cAMP level with forskolin and/or 3-isoburyl- 1 - methylxanthine(IBMX). Endogeneous follicular progesterone level increased rapidly in one hour of culture by treatment of frog pituitary homogenate(FPH) and reached peak level at 2 hours or later. But the absolute amount of progesterone produced (60-300 pg/follicle) or the peak time of the honnone level was different between individual animals. Basal level of progesterone in untreated sister follicles was very low (around 10 pg/follicle) and nearly undetectable in most cases regardless of culture lime. Secretion level of progesterone by the follicles obtained by measunng the honnone in the culture media was just the reflection of the intrafollicular level. Exogeneously added forskolin, an adenylate cyclase stimulator, and/or IBMX, a phosphodiesterase inhibitor, could mimic FPH action in terms of progesterone production and secretion. Thus, it seems clear that FPH regulates progesterone production via cAMP system in the follicle cells.

• Korean Journal of Animal Reproduction
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• v.14 no.4
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• pp.263-271
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• 1990

This study was carried out to investigate the factors affecting maturation in vitro of follicular oocytes in Korean Native Cattle. The bovine ovaries were obtained at a slaughter house and the follicular oocytes were recovered by aspirating the follicular fluid from the visible follicles of 3~6mm. The bovine oocytes were matured in vitro in TCM-199 containing FCS and hormones. The effects of TCM-199 salt type, number of oocytes per drop, incubation time and co-culture with granulosa cells on maturation of oocytes, were investigated. The results obtained are summarized as follows. 1. The maturation rates of follicular oocytes cultured for 22, 25 and 28 hours in Hank's TCM-199 or Earle's TCM-199 were 59.3, 59.6, 80% and 80.0, 90.0, 93.7%, respectively. The maturation rate of follicular oocytes in Earle's TCM-199 was faster and higher than in Hank's TCM-199(P<0.05). 2. The maturation rates of oocytes were 54.5, 62.5 and 62.0% when cultured the oocytes number 10, 20 and 40 per 200${mu}ell$ drop for 18 hours. 3. The maturation of follicular oocytes in the Korean Native Cattle was induced at 14 hours culture, by giving the maturation rate of 90.0% after 20 hours. 4. The maturation rates were 63.3% and 66.7%, respectigely when the oocytes were co-cultured with granulosa cells from medium-size follicles used immediately after recovery in the presence or absence of hormones added(0.02AU/ml FSH, 10$\mu\textrm{g}$/ml LH and 1$\mu\textrm{g}$/ml estradiol 17-$\beta$). When the oocytes were co-cultured with granulosa cells from medium-size follicles cultured for 3 days, the maturation rates were 20.8% and 76.2%, respectively(P<0.05). 5. The maturation rate were 88.0% and 70.0%, respectively when the oocytes were co-cultured with granulosa cells from large-size follicles used immediately after recovery in the presence or absence of hormones added(0.02AU/ml FSH, 10$\mu\textrm{g}$/ml LH and 1$\mu\textrm{g}$/ml estradiol 17-$\beta$)(P<0.05). When the hormones added(0.02AU/ml FSH, 10$\mu\textrm{g}$/ml LH and 1$\mu\textrm{g}$/ml estradiol 17-$\beta$)(P<0.05). When the oocytes were co-cultred with granulosa cells from large-size follicles cultured for 3 days, the maturation rates were 41.2% and 73.3%, respectively(P<0.05).