• Journal of Embryo Transfer
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• v.9 no.3
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• pp.243-247
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• 1994

최근 의약적으로 유용한 단백질을 대량 생산키 위한 실현 가능한 방법이 유전자변환 가축의 이용과 관련되어 발전되어 왔다. 이러한 유전자 변환동물은 이종의 단백질을 유즙속으로 분비시키는 생체반응기로서 이용되고 있다. 이러한 전략적 목적을 위해 현재 유전자 변환동물의 생산을 위한 이용에 있어 여러 가지 방법들이 보고되고 있다. 그러나 ES 세포의 사용이 이러한 방법들 사이에서 가장 실질적인 것으로 추정되고 있다. 본 실험에서는 유전자 구축을 위해 사람 황체 호르몬(human luteinizing hormone; hLH)의 전사를 유도하기 위해 각각 2.2 및 0.5 kb의 토끼 $\beta$-casein pronoter 단편을 이용하여 생쥐의 유선에 hLH를 발현시키도록 조절하고 발현이 thynidine kinase(TK) pronoter에 의해 좌우되는 neo 유전자를 selectable marker로서 plasnid속에 삽입하였다. 그 결과 생긴 구축 유전자는 각각 pCas 2.2와 pCas 0.5로 명명하였다. 구축된 유전자로 2$\times$107의 TT-2 ES세포를 170V, 550$\mu$F로 100$\mu$g의 선상 plasmid에 의해 electroporation 시켰다. 감염된 colony들은 250$\mu$g/$m\ell$ G418을 함유하는 ESM 배양액에서 선별 7일 이후에 회수하여 성공적으로 감염된 ES세포는 PCR 및 Southern blot에 의해 확인되었고 그들 중 나머지는 trypsin 처리 후 각각 미세조작과 공배양 기술을 사용하여 ICR 생쥐의 8세포기 수정란 속에 도입하였다. 결국 24시간 동안 37$^{\circ}C$, 5% $CO_2$에서 배양된 배반포를 chimera의 생산을 위해 위임신 유기된 G418 선발처리 이후 400 및 275개의 ES 세포 colony가 생존하였으며, 3개의 ES 세포으 colony 의 genome 속에 임의적으로 plamid가 삽입된 것을 Southern blot에 의해 확인되었다. 총 13 chimera 생쥐가 3 colony로부터 생산되었으나 germ-line chimera는 현재 조사중이다. chimera 생산빈도는 공배양 기술보다 주입방법에서 현저히 높았다.

• Journal of Animal Science and Technology
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• v.59 no.10
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• pp.21.1-21.6
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• 2017

Background: The aim of the study is to evaluate the effectiveness of thermographic monitoring, using the temperature changes of perianal and perivulvar areas for the determination of estrus in Anatolian Shepherd bitches. Fifteen bitches were used in the study. Blood and vaginal smear samples were collected and thermographic monitoring of perianal and perivulvar areas were carried out starting from proestrus to early diestrus. Also, external signs of estrus were investigated. Smear samples were evaluated by light microscopy after Diff-Quik staining method and superficial and keratinized superficial cells were determined as percentage (S + KS%). Progesterone and luteinizing hormone measurements were done by radioimmunoassay. The difference in temperature between perianal and perivulvar areas was evaluated through thermographic images by FLIR ResearchIR Software. Results: According to the results obtained from the study, differences between progesterone and S + KS% were statistically significant (P < 0,05). Although temperature showed increase and decrease with progesterone and S + KS%, the differences were not important statistically (P > 0,05). Serum luteinizing hormone levels did not sign any difference (P > 0,05). Conclusions: As a result, thermographic monitoring alone is not enough for estrus detection in Anatolian Shepherd bitches. However, it can be used to assist the actual estrus detection technique in terms of providing some foreknowledge by evaluating the differences in temperature.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.2
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• pp.91-96
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• 2002

Objective : To evaluate luteinizing hormone (LH) in patients with infertility and recurrent spontaneous abortion. Material and Method: LH was tested by solid phase immunoradiometric assay based on monoclonal and polyclonal anti-LH antibodies. Results: Of 100 infertile patients, the number of less than 5 mIU/ml, 5$\sim$10 mIU/ml, greater than 10 mIU/ml in LH level was 67 (67%), 22 (22%), 11 (11%), respectively. Of 100 patients with recurrent spontaneous abortion, the number of less than 5 mIU/ml, 5$\sim$10 mIU/ml, greater than 10 mIU/ml in LH level was 79 (79%), 18 (18%), 3 (3%), respectively. There was a significant difference between patients with infertility and recurrent spontaneous abortion only in the group with LH level greater than 10 mIU/ml (p=.0325). Conclusions: High LH in the follicular phase is known to decrease pregnancy rate and increase abortion rate. But in this study the incidence of high LH in patients with recurrent spontaneous abortion was low. On the contrary, there was a significant increase of LH in infertile patients. So recurrent spontaneous abortion does not seem to be related to high LH level.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.3
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• pp.383-387
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• 1999

Objective: There is increasing evidence for the expression of rat in gene in several extrapituitary sites including testis and ovary. We also have demonstrated that the local LH expression in the rat epididymis and uterus, the major accessory sex organs in male and female reproductive system, respectively. Design: The present study was undertaken to elucidate whether the gene for LH receptor is expressed in rat uterus and whether the expressions of uterine LH and its receptor are differentially regulated during estrous cycle. Presence of the transcripts for rat LH receptor in the rat uterine tissue were confirmed by touchdown reverse transcription-polymerase chain reaction (RT-PCR). Results: In $LH{\beta}$ semi-quantitative RT-PCR, the highest expression level was shown in estrus stage. The level of ill receptor transcripts was also fluctuated during estrous cycle. In ovariectomized rats (OVX + Oil), the expressions of both uterine LH and LH-R were markedly reduced when compared to those from normal rats. Supplement with estradiol $17{\beta}$ to the ovariectomized rats (OVX + $E_2$) restored the expression levels of LH and its receptor to the levels in uteri from normal rats. Conclusion: Our findings indicated that 1) LH and its receptor gene are expressed in the rat uterus from cycling rats, 2) the expression of uterine LH and its receptor is mainly, if not all, under the control of ovarian sex steroid(s). These results suggested that the uterine LH may act as a local regulator with auto and/or paracrine manner, though the posibility that the pituitary LH may act directly on the regulation of uterine functions could not be discarded.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.52-52
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• 2003

For the evaluation of hormonal control of spermiation in fish, a method to quanify the spermiation response or mature Rhynchocypris oxycephalus (Sauvage and Dabry) to hormonal therapy is described. Spermatocrit was determined after 7 min centrifugation at 18,000 $\times$ｇ and sperm density was estimated by a standard hemocytormer method. Sperm density can be predicted from spermatocrit since their relationship is linear as described by the regression equation, Y=3.68X-27,18 ($R^2$=0.82, N=50), where Y is spermatocrit and X is sperm density. Milt production by mature R. oxycephalus was highest at 24 h after injection of 1,000 IU human chorionic gonadotropin (HCG) and 50 $\mu\textrm{g}$ luteinizing hormone-releasing hormone analogue (LHRHa) per kg body weight. Increased milt production coincided with low spermotocrit and sperm density levels. These results demonstrate that spermition in mature R. oxycephalus can be reliably evaluated by a spermatocrit method and that HCG and LHRHa are effecove in stimulating of spermiation in this species.

• Molecular & Cellular Toxicology
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• v.4 no.3
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• pp.230-234
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• 2008

Epidemiological studies demonstrate an adverse effect of several environmental and occupational exposures on male sex hormone. Bisphenol A (BPA) is a weak estrogen and a widely used industrial chemical. Epoxy resin painters may be continuously exposed to BPA at high concentrations. The effect of occupational exposure of BPA on male reproduction was examined by measuring the urinary BPA, testosterone and gonadotropic hormones of epoxy resin painters in the shipyard. The painters had significantly higher concentrations of urinary BPA (2.61${\pm}$1.08 ${\mu}g$/g creatinine) than controls (1.38${\pm}$0.5 9 ${\mu}g$/g creatinine). In serum, the testosterone level of painters was significantly decreased but the luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels of painters were significantly higher than controls. Occupational exposure to BPA influences testosterone and gonadotropic hormones in male workers.

• Journal of Embryo Transfer
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• v.33 no.4
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• pp.271-280
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• 2018

Here, we evaluated the mode of programmed cell death during porcine oocyte maturation by comparing the two major pathways associated with programmed cell death, apoptosis (type I), and autophagy (type II). We investigated the expression and localization of major genes involved in autophagy and apoptosis at mRNA and protein levels. Furthermore, the effect of hormonal stimulation on autophagy and apoptosis was analyzed. We found that the activity of autophagy-associated genes was increased in the cumulus-oocyte complexes (COCs) following follicle-stimulating hormone (FSH) treatment, while the addition of luteinizing hormone (LH) reversed this effect. The expression of proteins associated with autophagy was the highest in FSH-treated COCs. On the other hand, caspase-3 protein level was maximum in COCs cultured with LH. The treatment with rapamycin resulted in the effect similar to that observed with FSH treatment and increased autophagy activity. Thus, hormonal stimulation of pig oocytes resulted in distinct patterns of maturation. The high-quality oocytes majorly relied on the type II pathway (autophagy), while the type I pathway (apoptosis) was more prominent among poor-quality oocytes. Further investigation of this distinction may allow the development of techniques to produce high-quality oocytes in porcine in vitro fertilization.

• The Journal of Korean Obstetrics and Gynecology
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• v.36 no.2
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• pp.1-19
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• 2023

Objectives: The purpose of this health promotion project for Korean medicine is to promote the health of climacteric women in Boeun-gun. Methods: During the project, pressure needle acupuncture treatment, herbal medicine treatment, meditation pore therapy, and health education were conducted. To evaluate the results, basic health surveys (body height, body composition test, blood pressure etc.), female hormone tests, Kupperman's index (KI), Menopause-specific quality of life questionnaire (MENQOL), and Perceived stress scales (PSS) were conducted. Results: No significant change was observed in the body composition test and blood pressure after the project. Follicle stimulating hormone (FSH) was significantly increased after the project, but Luteinizing Hormone (LH) and total estrogen levels were not significantly changed. KI and MENQOL scores significantly decreased after the project, and significantly decreased in 4 out of 11 items of KI and 3 out of 4 domains of MEMQOL. No significant change was observed in the PSS score after the project. Conclusions: From the results of this project, it can be seen that the Korean medicine health promotion project can help alleviate symptoms of climacteric syndrome and postmenopausal syndrome.

• Animal cells and systems
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• v.1 no.4
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• pp.595-602
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• 1997

Treatment of newborn female rats with gonadal steroids induces permanent sterility in adulthood. We investigated the alteration in expression patterns of gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) in neonatally estrogenized sterile rats (ESR). Newborn female rats received daily injections of 17${\beta}$-estradiol (E, 10 ${\mu}$g) from the day of birth (day 1) to postnatal day 5. Controls were subjected to vehicles over the same period. All animals were sacrificed on week 7 after birth. Hypothalamic GnRH mANA levels were markedly higher in all ESR than in controls, while hypothalamic GnRH contents in ESR increased in proportion to the frequency of daily administration of E. However, both pituitary LH6 mRNA and serum LH levels were inversely decreased by the same treatment. The data indicate that neonatal exposure of E equally elevates the expression of GnRH gene, but reduces the secretion of GnRH, accordingly leading to attenuation of LH6 gene expression and circulating LH levels. The temporal effect of E and/or progesterone (P) on GnRH and LH6 mRNA levels was also examined in ESR. Newborn female rats were daily injected with E (10 ${\mu}$g) or vehicle for five successive days from day 1 and ovariectomized at week 5. They were implanted with E (235 ${\mu}$g/ml) two days prior to week 7, injected with P (1 mg) 42 h later, and sacrificed 7 h after P administration. In ovariectomized controls, hypothalamic GnRH mRNA levels were dropped to half by treatment of E and restored by subsequent treatment of P. The negative feedback action of E on GnRH mRNA levels observed in ovariectomized rats was completely blocked by neonatal exposure of E. The change in pituitary LH mRNA levels was similar to that in hypothalamic GnRH mRNA levels. Taken together, the results suggest that neonatal treatment of E alters the synthesis and release of GnRH in adulthood and furthermore blocks the negative feedback regulation of E which occurs normally after ovariectomy.

• Clinical and Experimental Reproductive Medicine
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• v.15 no.2
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• pp.135-147
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• 1988

To compare the stimulation effect of the ratio in follicle stimulating hormone and luteinizing hormone in induction of multiple follicular growth, the serum $E_2$ level, the diameter of follicle, number of aspirated follicles and cleavage rate of in vitro fertilized preovulatory oocytes as well as the pregnancy rate were evaluated. Forty one patients with irreparable tubal disease were stimulated by hMG(n=24) or FSH/hMG(n=17) for the purpose of in vitro fertilization and embryo transfer. The following results were obtained. 1. Serum estradiol($E_2$) levels on the day of hCG administration were $921.0{\pm}353.3\;pg/ml$ in hMG group and $1272.9{\pm}1060.6\;pg/ml$ in FSH/hMG group. The serum $E_2$ value of hMG group was significantly lower than that of FSH/hMG group. 2. The diameter of leading follicle by ultrasonogram on the day of hCG administration were $16.2{\pm}2.0\;mm$ in hMG group and $16.2{\pm}2.6\;mm$ in FSH/hMG group. No significant difference of follicle diameter between two groups was demonstrated. 3. The number of follicles with diameter above 10 mm by sonogram on the day of hCG injection were $3.91{\pm}2.32$ in hMG group and $6.52{\pm}3.86$ in FSH/hMG group. There was significant difference of number of follicles between two groups, (p< 0.01). 4. The number of oocytes found per patient at aspiration were $2.59{\pm}1.00$ in hMG group and 3. $76{\pm}2.31$ in FSH/hMG group. There was significant difference of number of aspirated oocytes between two groups. (p< 0.05). 5. The detection rate of preovulatory oocyte at aspiration were 68.4%(39/57) in hMG group (n=22) and 77.6%(38/49) in FSH/hMG group (n=13). 6. The cleavage rate of preovulatory oocyte at 44 hours after insemination were 74.4%(29/39) in hMG group(n=22) and 81.6%(31/38) in FSH/hMG group (n=13). When only hMG was used, one pregnancy was established in 15 patients to whom 29 zygotes were transferred. And a full term normal female baby was delivered by elective cesarean section. In the FSH/hMG group, five pregnancies out of 9 transferred patients were confirmed by serum ${\beta}-hCG$. Two pregnancies were spontaneously aborted before the 6th week of pregnancy. One patient aborted her baby at the 18th week of pregnancy because of incompetent internal os of the cervix. Two patients delivered two full term babies by elective cesarean section. From the above findings, paralell with the increase in the ratio of exogenous follicle stimulating hormone to luteinizing hormone, an increase in oocyte recovery was observed as well as an improvements in pregnancy rate. It was concluded that FSH enrichment early in the follicular phase had a beneficial effect in the controlled ovarian hyperstimulation.