• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.2
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• pp.428-435
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• 2006

Cortex Mori radicis has been used as a components of antidiabetics, antiasthma and diuresis in Oriental Medicine. This experiment examined the effect of an extracts, obtained from the acetone(CM-A) and aqueous(CM-W) extracts of Cortex Mori radicis, on hair growing activity of the C57BL/6N mice after topical application to skin. We investigated the number of hair follicle and mast cells, and changes of subpopulation of splenocytes and thymocytes in skin for 16 day. The results were as follows : The Hair growing effect in experimental groups was more increased in 85%(CM-A) and 90%(CM-W) than control group(10%) in hair depilated area. The number of hair follicle in experimental groups(CM-A and CM-W) was more increased than control group. Splenic B/T lymphocytes of CM-A group were decreased compare to control group. CD4/CD8 positive TH cells in splenic T lymphocytes of CM-W group were increased compare to control group. These results suggest that CM-A and CM-W may be used in treatment of alopecia areata.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.5
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• pp.632-636
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• 2005

This study was conducted to investigate the effects of removal of all ovarian follicles through repeated transvaginal follicle aspiration (TVFA) on the subsequent follicular populations in buffaloes. This information is crucial for determining the optimum time interval between successive aspirations for recovering oocytes from live buffaloes through Transvaginal Oocyte Retrieval (TVOR). The oestrus of cycling buffaloes (n=5) were synchronized by a single PGF injection schedule. All the ovarian follicles were removed once every 7 days for 6 weeks through TVFA, starting from Day 7 of the oestrous cycle (Day 0 = day of oestrus). The number and size of individual ovarian follicles was recorded at Day 3 and Day 5 (Day 0 = day of TVFA) through transrectal ultrasonography. The follicles were classified on the basis of their diameter as small (3-5 mm), medium (6-9 mm) and large ($\geq$10 mm). There was no difference in the number of small and medium follicles, and the number of total follicles between Day 3 and Day 5. However, the number of large follicles was significantly higher (p<0.05) at Day 5 than that at Day 3. There was a significant (p<0.05) decrease in the proportion of small follicles and an increase (p<0.05) in the proportion of large follicles from Day 3 to Day 5, with no change in the proportion of medium follicles. The number of total follicles at Day 3 or Day 5 did not differ during the 6 TVFA sessions. It can be concluded that an interval of 3 days is more suitable than that of 5 days between successive aspirations for recovering oocytes through TVOR in a twice weekly schedule and that repeated removal of follicles through TVFA does not adversely affect the number of total follicles 3 or 5 days after TVFA.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.2
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• pp.168-173
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• 2009

Human follicle-stimulating hormone (hFSH) is a pituitary glycoprotein that regulates follicular development and ovulation. Clinically, hFSH has been used to induce follicular growth in infertile women. The hormone is composed of heterodimers, including a common ${\alpha}$ subunit among the gonadotropin family and a hormone-specific ${\beta}$ subunit. Since assembly of the heterodimer is a rate-limiting step in the production of functional hFSH, transgenic clone cows carrying a single-chain hFSH transgene may efficiently produce functional hormone. Genes encoding the ${\alpha}$ and ${\beta}$ subunits of hFSH were linked using the C-terminal peptide sequence from the ${\beta}$ subunit of human chorionic gonadotropin. Bovine fetal fibroblasts were transfected with the gene construct, including the goat ${\beta}$-casein promoter and a single-chain hFSH coding sequence. Transfected fibroblasts were transferred into enucleated oocytes, and individual nuclear transfer (NT) embryos developed to the blastocyst stage were analyzed for the transgene by polymerase chain reaction. Seventy eight blastocysts (30.8%) were developed from 259 reconstructed embryos. Among these blastocysts, the hFSH gene was detected in 70.8% (34/48) of the embryos. Subsequent transfer of hFSH-transgenic clone embryos to 31 recipients results in 11 (35.5%) early pregnancies. However, all fetuses were lost before reaching day 180 of gestation. The results from this study demonstrated that bovine NT embryos carrying single-chain hFSH could be produced, and further extensive studies in which NT embryos are transferred to more recipients may give rise to single chain hFSH-transgenic cows for biomedical applications.

• Toxicological Research
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• v.29 no.4
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• pp.241-247
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• 2013

This study was carried out to examine the action mechanism of Chamaecyparis obtusa oil (CO) on hair growth in C57BL/6 mice. For alkaline phosphatase (ALP) and ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GT) activities in the skin tissue, at week 4, the 3% minoxidil (MXD) and 3% CO treatment groups showed an ALP activity that was significantly higher by 85% (p < 0.001) and 48% (p < 0.05) and an ${\gamma}$-GT activity that was significantly higher by 294% (p < 0.01) and 254% (p < 0.05) respectively, as compared to the saline (SA) treatment group. For insulin-like growth factor-1 (IGF-1) mRNA expression in the skin tissue, at week 4, the MXD and CO groups showed a significantly higher expression by 204% (p < 0.05) and 426% (p < 0.01) respectively, as compared to the SA group. At week 4, vascular endothelial growth factor (VEGF) expression in the MXD and CO groups showed a significantly higher expression by 74% and 96% (p < 0.05) respectively, however, epidermal growth factor (EGF) expression in the MXD and CO groups showed a significantly lower expression by 66% and 61% (p < 0.05) respectively, as compared to the SA group. Stem cell factor (SCF) expression in the MXD and CO groups was observed by immunohis-tochemistry as significant in a part of the bulge around the hair follicle and in a part of the basal layer of the epidermis. Taking all the results together, on the basis of effects on ALP and ${\gamma}$-GT activity, and the expression of IGF-1, VEGF and SCF, which are related to the promotion of hair growth, it can be concluded that CO induced a proliferation and division of hair follicle cells and maintained the anagen phase. Because EGF expression was decreased significantly, CO could delay the transition to the catagen phase.

• Journal of Embryo Transfer
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• v.15 no.1
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• pp.1-8
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• 2000

The ultrasound-guided oocytes cllection (ovum pick-up ; OPU) has become a substitution for superovlation in cattle. The objective of this study was to examine the effect of OPU frequency on the in vitro production of embryos in Hanwoo cattle. Six cycling Hanwoo cows were distributed into two groups for either once or twice weekly OPU sessions. Oocytes were collected by ultrasound-guided follicle aspiration(SA600) using a 6.5HMz transducer and attached with 18 gauge needle, with vacuum pressure of 40 mmHg. The cumulus-oocyte complexes (COCs) collected from each donor were matured in TCM 199 supplemented with 10% fetal bovine serum at 5% CO2 in air at 38.5$^{\circ}C$ for 22h and in vitro matured oocytes were co-incubated with sperm(separated by Percoll gradient) for 6h. The zygotes were co-cultured on cumulus cell monolayer in 10ul droplets in the same culture medium and conditions used for IVM for 7 days. On Day 7 of culture, development to blastocysts was examined. Although the number of oocytes collected was variable depending on individuals, overall embryo production in the twice per week OPU sessions was better that in the once per week sessions(6~21 vs 2~7 blastocysts produced, respectively). Two cows(E, A) were good oocyte donors and embryo production was superior in cow C ; however, cow F was a poor donor as compared to the others. In conclusion, these results suggest that for embryo production, twice weekly OPU sessions were better than once per week for producing embryos in vitro from Hanwoo cattle.

• Korean Journal of Clinical Laboratory Science
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• v.40 no.1
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• pp.31-40
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• 2008

Despite of the importance of the primordial follicle (PMF) recruitment, factors and mechanisms for process are poorly understood. To evaluate expression and role of the follicular transition from PMF to PMF/primary follicles (PMIF) in the present study, we evaluated expression of lats1, lats2, cyclin A1, and cyclin A2 mRNA and protein, and elucidated and role of lats1-cyclin A in the follicular transition from PMF to PRIF. To analysis of differential expression in PMF and PMIF, each stage follicles were collected by day1 and day5 of immuno-compromised rats (ICR) and analyzed by real-time PCR for the genes. For localization of mRNAs and proteins of the genes, in situ hybridization and immunohistochemistry were performed. We confirmed that the lats1, lats2, cyclin A1, and cyclin A2 mRNA were more expressed in PMF than PMIF. Localization of the four genes expression were observed in nuclei of oocytes from the arrested primordial, and in the surrounding granulosa cells of the growing follicles. The mRNA expressions were gradually decreased with follicular development. From immunohistochemistry studies, Cyclin A1 protein expression were observed in oocyte cytoplasmas of early stage follicles, while observed in granulose cells and oocyte nucleoli during growing follicles. This study suggested that the presence of lats gene family might perform negatively regulation of cell proliferation by modulation of the CDC2/Cyclin A complex activity. lats-cyclin A genes in oocytes of the early stage follicles might play a role in the meiotic cell cycle arrest of the primary oocytes at the primordial follicle stage as well as the follicular growth.

• Korean Journal of Medicinal Crop Science
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• v.22 no.2
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• pp.113-120
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• 2014

These studies were conducted to discriminate Cynanchum wilfordii, C. auriculatum, Metaplexis japonica, and Polygonum multiflorum by morphological characters. Cynanchum wilfordii, C. auriculatum, and Metaplexis japonica(Asclepiadaceae) were easily distinguished from Polygonum multiflorum by twining stem to the left, producing opposite leaves, having laticiferous stems and leaves at cutting cross-sectionally, and lacking lignification of the stem. Four species had different morphological characters of flower. Comparing four species to fruit type, three species of Asclepiadaceae had follicle, while Polygonum multiflorum had achene with three wings. Follicle shape was lanceolate, narrowly lanceolate, and widely lanceolate in C. wilfordii, C. auriculatum, and Metaplexis japonica, respectively. Whereas seed color of four species was dark brown similarly, seed shape and 1,000 seed weight were useful characters to discrimate between three species of Asclepiadaceae and Polygonum multiflorum. Shape and color of root were thickened and yellowish brown in C. wilfordii and C. auriculatum. They were thin and long and white in Metaplexis japonica, and fusiform and reddish brown in Polygonum multiflorum. They would be useful characters to distinguish Cynanchum spp., Metaplexis japonica, and Polygonum multiflorum, however, impossible characters to discriminate between C. wilfordii and C. auriculatum.

• Reproductive and Developmental Biology
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• v.33 no.4
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• pp.189-194
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• 2009

The objective of this study was to determine effects of different culture media. Preantral follicles were mechanically extracted from bovine ovaries and cultured for 16 days in tissue culture medium (TCM)-199, DMEM or alpha-minimal essential medium ($\alpha$-MEM) + 10% FBS + 0.1 mg/ml sodium pyruvate + 100 mIU/ml FSH. The collected primary follicles from ovary were higher than the primary and secondary follicles. The survival rates of the follicles in TCM-199 were significantly higher (p<0.05) than those in DMEM and $\alpha$-MEM. The diameter of the follicles progressively increased during 12 days of culture. The maximum size ($139.1{\pm}5.4\;{\mu}m$) reached on Day 12 of the in vitro culture and decreased on Day 16. These results suggest that in a culture of bovine preantral follicles, TCM-199 is an optimal medium and a longer-term culture of preantral follicles (>12 days) may be needed to form antra.

• Proceedings of the Korea Society of Environmental Biology Conference
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• 2002.11a
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• pp.123-126
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• 2002

Pollutants that are persistent, bioaccurnulative, and toxic have been linked to numerous adverse effects in human and animals, PBTs include heavy metals, polychlorinated biphenyls (PCBs), dioxins, polycyclic aromatic compounds (PACs) in addition to pesticides. This study focuses on toxic effects of the PBTs except pesticides on insects. Eight PBTs were selected from subgroups: three heavy metals (Pb, Hg, and Cd), two PCB mixtures (Aroclor mixtures 1 and 2), 2,3,7,8-tetrachlorodibenzo-p-dioxin, two monophenols (4-octylphenol and 4-nonylphenol), and tetrabutyltin, Beet armyworm, Spodoptera exigua, was used as test target insect species. Three physiological markers (metamorphosis, immune reaction, and follicle patency) were assessed in each exposure to different doses of the PCBs. Heat-shock proteins as molecular markers were also analyzed in response to the PCBs. All tested PBTs were toxic to metamorphosis from larvae to pupae when they were applied with diet. Two PCB mixtures were the most toxic compounds in this assay by giving significant toxicity at 0.005 ppm, while others had from 10 to 1000 ppm. Dioxin (0.1 ppb), tetrabutyltin (0.1 ppb), Pb (10 ppb), and Hg (0,01 ppb) were potent to inhibit immune reactions analyzed by inducing phenoloxidase activity and blocked phospholipase $A_2$ enzyme, Tetrabutyltin and dioxin significantly induced follicle cell patency, but their effects were lower than that of endogenous juvenile hormone, Dioxin, Pb, Hg, and Cd could induce the expression of heat shock proteins that were detected by immunoblotting against human HSP70 monoclonal antibody. HSP78 and HSP80 were upregulated in response to the PBTs. This expression was detected from the fat body and epidermis at as fast as 4h after injection. All these results clearly suggest that PBTs give significant ecotoxicity to insects that are valuable organisms in our environment.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.395-402
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• 2005

Biologically active recombinant human follicle stimulating hormone (rhFSH) was produced in Chinese hamster ovary cells and purified by a series of chromatographic steps. The chromatographic steps included anion-exchange chromatography (DEAE Sepharose F/F, Q Sepharose F/F), hydrophobic interaction chromatography (Source 15 PHE), and hydroxyapatite chromatography (Macro-Prep ceramic hydroxyapatite type I). A distinctive step of the purification process developed was the use of ZnCl$_2$ for the removal of non-glycosylated or lowly-glycosylated FSH and impurities through co-precipitation with Zn$^{2+}$. Purified rhFSH was identified and characterized by several physicochemical and biological methods such as gel electrophoresis, high-performance liquid chromatography, amino acid analysis, carbohydrate analysis, and biological activity. The overall yield of the purification was ~$30\%$. The rhFSH preparation obtained showed high purity (>$99\%$) and high in vivo potency (>16,000 IU/mg). Carbohydrate analysis suggested that the purified rhFSH contained approximately $40\%$ (w/w) carbohydrate with di­or tri-antennary structure on average, which is somewhat more heavily sialylated than commercially available rhFSH. In conclusion, the results of these analyses established an identity of the purified rhFSH with natural FSH from human pituitary glands, and furthermore, the purified rhFSH preparation showed higher in vivo potency and was slightly more heavily sialylated than commercially available rhFSH.