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Improvement of Pregnancy Rates by Coculture of Human Embryos with Cumulus Cells in Glucose and Phosphate Free M-TALP Media (Glucose와 Phosphate가 제거된 M-TALP 배지에서의 난구세포 공배양에 의한 임신율 향상에 관한 연구)

  • Chung, B.S.;Chang, W.H.;Lee, M.H.;Kim, J.Y.;Bang, J.H.;Kim, K.H.;Suh, T.K.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.75-81
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    • 1999
  • The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.

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Chromosomal Abnormalities in Human Oocytes Fail to Fertilize after Insemination In Vitro (수정에 실패한 인간 난자에 있어서의 염색체의 수의 이상)

  • Son, Weon-Young;Lee, Kyung-Ah;Park, Sang-Hee;Han, Sei-Yul;Yoon, Tae-Ki;Jung, Hyung-Min;Kwak, In-Pyung;Cha, Kwang-Yul
    • Clinical and Experimental Reproductive Medicine
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    • v.22 no.2
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    • pp.203-210
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    • 1995
  • Many oocytes fail to fertilize and cleave in vitro and many embryos transferred back to uterus fail to implant or maintain implantation. Chromosomal abnormalities in the male and female gametes may contribute to this loss. The higher incidence of meiotic chromosomal abnormalities bas been found in oocytes than in sperm. The wide range of incidence of chromosomal abnormalities in unfertilized oocytes has been reported in human IVF program (26-63%). However, factors affecting chromosomal abnormalities are not well understood. The present study has been conducted to investigate effects of the method for ovarian hyperstimulation, women's age, and the number of oocytes retrieved per patients on the incidence of numerical chromosomal abnormalities. Five hundred eighty four unfertilized metaphase II oocytes were subjected to chromosomal analysis. Included unfertilized oocytes were from 220 patients (mean $age=32.7{\pm}3.0$) and three hundred thirty oocytes were legible for analysis. Two hundred fourty five oocytes out of 330 (73.3%) were normal, while 38 (11.5%) were hyperploidy, 35 (10.6%) were hypoploidy, and 12 (3.6%) were diploidy. Significant difference in chromosomal abnormalities was not found between two patient groups stimulated by follicular stimulating hormone/human menopausal gonadotrophin (FSH/HMG) (25.9%) and gonadotrophin-releasing hormone agonist/follicular stimulating hormone/human menopausal gonadotrophin (GnRHa/FSH/HMG) (28%). There was a tendency of increasing chromosomal abnormalities in unfertilized oocytes from older patients (<30 yrs: 20.3%, 30-34yrs: 26.9%, >34 yrs: 35.3%). The number of oocytes retrieved per patient had no effect the incidence of chromosomal abnormalities (1-5: 31. 4%, 6-10: 29.8%, 11-15: 28.6%, > 15: 16.5%). These results from the present study suggest that the chromosomal abnormalities observed in the unfertilized oocytes has not affected by the stimulation methods, patient's age, and the number of oocytes retrieved per patients.

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A Comparative Study on Clinical Effectiveness of Human Chorionic Gonadotropin and Progesterone on Luteal Support in Controlled Ovarian Hyperstimulatian far IVF (체외수정 시술시 과배란 유도에서 Luteal Phase Support에 Human Chorionic Gonadotropin과 Progesterone의 효용성에 관한 비교 연구)

  • Nah, O-Soon;Lee, Sang-Hoon;Bae, Do-Whan
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.3
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    • pp.233-240
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    • 1994
  • This study was conducted to compare the endocrine milieu, and pregnancy rates in In Vitro Fertilization and Embryo Transfer(IVF-ET) program employing combined with gonadotropin releasing hormone agonist(GnRH-a) and pergonal(LH 75lU+FSH 75lU) when either human chorionic gonadotropin(HCG) or progesterone were used for luteal phase support. A total number of 40 IVF-ET treatment cycles were prospectively studied. Ovarian hyperstimulation method was modified ultrashort protocol using GnRH-a. All patients started Decapeptyl at menstrual cycle day # 2, and HMG was started at # 3 days. When leading follicle was ${\geqq}$18mm or at least two follicles were ${\geqq}$14mm in diameter, HCG 10000lU intramuscularly was injected. After 36 hours HCG administration, oocytes were retrieved as usual guided by transvaginal ultrasound. Embryo were transfered 36-48 hours later. The patient's cycles were prospectively randomized to receive HCG(20cycles) or Progesterone (20cycles) for luteal support. The progesterone group received 25mg 1M starting from the day of ET. The HCG group received 1500IU 1M. on days 0, +2, +5 after ET. Estadiol($E_2$) and Progesterone($P_4$) were measured on the day of oocyte aspiration, ET day, and every 6 days thereafter. Results were follows as; 1. Estradiol, progesterone and LH levels on the day of HCG trigger, retrieved oocytes and number of transfered embryo were not significantly different in both groups. 2. On the day of aspiration and embryo transfered day, $E_2$, $P_4$ level were significantly higher in progesterone group than HCG group(p<0.01). 3. $E_2$, $P_4$ level on 6 days after ET were significantly higher in progesterone group than HCG group(p<0.01). But, $P_4/E_2$ ratio was not different in both groups. 4. $E_2$, $P_4$ level 12 days after ET were decreased abruptly in both groups and higher hormonal level appeared in HCG group(P<0.01). 5. The total pregnancy rate in the HCG group was 40% (8/20) and in the progesterone group 15%(3/20). 6. Comparing the pregnant and nonpregnant cases progesterone group was not different the hormonal status. In HCG group, pregnant cases appeared in higher $P_4$, $P_4/E_2$ ratio than nonpregnanct cases(P<0.01).

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Growth Rate of Dominant Follicles During Controlled Ovarian Hyperstimulation (과배란유도 여성에서 우성난포의 성장속도)

  • Hwang, Eun-Ju;Ku, Seung-Yup;Kim, Yong-Jin;Jee, Byung-Chul;Suh, Chang-Suk;Kim, Seok-Hyun;Choi, Young-Min;Kim, Jung-Gu;Moon, Shin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.3
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    • pp.253-259
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    • 2010
  • Objective: To evaluate if there is any correlation between the growth rate of dominant follicles and clinical characteristics or outcome variables in women undergoing controlled ovarian hyperstimulation (COH). Methods: This study was performed in 313 in vitro fertilization (IVF) cycles. Follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) were measured on day 3 of menstrual cycle, and serial ultrasonographic measurement of the diameter of growing follicles was performed. The growth rates of dominant follicles calculated by diameter difference divided by days were correlated with clinical characteristics and outcome variables. Results: There was no significant difference in the growth rate of the dominant follicles between gonadotropin releasing hormone (GnRH) agonist and antagonist cycles. No significant correlation was found between the growth rates and evaluated factors such as age, body mass index, LH, FSH, $E_2$, retrieved oocytes and fertilization rate. Conclusion: The Growth rate of dominant follicles seems to show an independent feature of basal characteristics and ovarian response.

Comparison of follitropin ${\beta}$ administered by a pen device with follitropin ${\beta}$ administered by a conventional syringe in patients undergoing IVF-ET

  • Kang, Hyuk-Jae;Kim, Chung-Hoon;Ahn, Jun-Woo;Lee, Hyang-Ah;Kim, Sung-Hoon;Chae, Hee-Dong;Kang, Byung-Moon
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.1
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    • pp.37-41
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    • 2011
  • Objective: To compare the effectiveness and convenience of a pen device for the self-administration of follitropin ${\beta}$ with a conventional syringe delivering follitropin ${\beta}$ solution in patients undergoing IVF-ET. Methods: GnRH agonist long protocol was used for controlled ovarian stimulation (COS) in all subjects. A total of 100 patients were randomized into the pen device group or the conventional syringe group on the first day of COS. Local tolerance reactions were assessed within 5 minutes, at 1 hour and at 3 hours after each injection. On the day of hCG injection, patients were asked to rate their overall pain and convenience experienced with self-injection on a visual anlaogue scale (VAS). Results: There were no differences in patients' characteristics between the two groups. The duration of COS was significantly shorter in the pen device group than in the conventional syringe group. Patients included in the pen device group needed a significantly smaller amount of follitropin ${\beta}$. However, no differences between the two groups were found in IVF results and pregnancy outcome. The incidence of local pain within 5 minutes, at 1 hour and at 3 hours after the injection was significantly lower in the pen device group. VAS scores indicated that injections using the pen device were significantly less painful and more convenient. Conclusion: The pen device for self-administration of follitropin ${\beta}$ is less painful, safer and more convenient for the patients, and can be more effective because of the shorter duration and smaller dose of follitropin ${\beta}$ when compared with the conventional syringe.

Seven-Day Continuous Abstinence Rate from Smoking at 1, 2, or 3 Years after the Use of Varenicline

  • Kim, Jin Se;Jang, Ju Young;Park, Eun Hye;Lee, Joo Young;Gu, Kang Mo;Jung, Jae Woo;Choi, Jae Chol;Shin, Jong Wook;Park, In Won;Choi, Byoung Whui;Kim, Jae Yeol
    • Tuberculosis and Respiratory Diseases
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    • v.78 no.2
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    • pp.92-98
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    • 2015
  • Background: Varenicline, a selective partial agonist/antagonist of the ${\alpha}4{\beta}2$ nicotinic receptor, has proven effectiveness for smoking cessation by several randomized, controlled trials. Because few studies have evaluated the long-term efficacy of varenicline, we tried to evaluate the smoking status of varenicline users up to 3 years after the initial prescription of the drug. Methods: We interviewed varenicline users who were prescribed the drug from June 2007 to May 2010 by telephone, from June 2010 to May 2011. Results: One-hundred and thirty-three of 250 varenicline users (53.2%) were available for the survey. Seven-day continuous abstinence from smoking was adhered to by 17 of 39 respondents (43.6%) at 1 year, and 11 of 36 (30.6%) and 19 of 58 (32.8%) at 2 and 3 years since the first use of varenicline, respectively. Compared to current smokers, successful quitters were older (55.0 years vs. 49.9 years, p=0.01), had better compliance to the 12-week course (27.7 vs. 9.3%, p=0.01), and had taken varenicline longer (10.1 vs. 5.9 weeks, p=0.01). Fifty-four of 71 current smokers (76.1%) were willing to stop smoking in the near future. The preferred ways to cease smoking were will-power (48.1%), varenicline (25.9%), nicotine replacement therapy (11.1%), and others (14.9%). Conclusion: Smokers should be encouraged to stick to the proven way for recommended period of time for successful cessation of smoking.

Effects of Morphine and Buprenorphine Intravenous Infusion for Postoperative Pain (Morphine과 Buprenorphine의 지속적 정주에 의한 술후 통증 억제효과의 비교)

  • Baek, Sun-Ki;So, Sang-Yoon;Han, Young-Jin;Choe, Huhn
    • The Korean Journal of Pain
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    • v.7 no.1
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    • pp.78-83
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    • 1994
  • Morphine, a $\mu$-receptor agonist, produces strong analgesic effect with some side effects such as nausea, vomiting, urinary retension, somnolence, and respiratory depression. Buprenorphine also provides strong analgesic effects, and hemodynamic changes after continuous infusion of morphine, or buprenorphine-ketorolac combination in gynecologic patients. Analgesic effect was assessed by visual analogue scale(VAS) and Prince Henry scale(PHS). Morphine group, initial 2 mg of bolus morphine was followed by 48 mg/96 ml of continuous infusion. Buprenorphine group, initial 0.1 mg of buprenorphine was followed up with infusion by 2.3 mg/100 ml. Half dose of both initial bolus and maintenance buprenorphine with ketorolac 15 mg for bolus and 60 mg for maintenance were infused in buprenorphine-ketorolac combination group. No significant hemodynamic changes were seen in any of the groups. VAS significantly decreased after 15 minutes of infusion in all three groups, and was significantly lower in morphine group than the other 2 groups, after 60 minutes. PHS was decreased significantly 15 minutes after infusion in the morphine group, and after 60 minutes in two other groups. The incidence of side effects were similar between morphine and buprenorphine groups, but significantly reduced in buprenorphine-ketorolac combination group. We concluded that buprenorphine could be an alternative to morphine for postoperative pain, and smaller dose of buprenorphine could be used if it is combined with ketorolac with less side effects.

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Signal Transduction of Eel Luteinizing Hormone Receptor (eelLHR) and Follicle Stimulating Hormone Receptor (eelFSHR) by Recombinant Equine Chorionic Gonadotropin (rec-eCG) and Native eCG

  • Byambaragchaa, Munkhzaya;Lee, So-Yun;Kim, Dae-Jung;Kang, Myung-Hwa;Min, Kwan-Sik
    • Development and Reproduction
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    • v.22 no.1
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    • pp.55-64
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    • 2018
  • Previous studies showed that recombinant equine chorionic gonadotropin ($rec-eCG{\beta}/{\alpha}$) exhibits both follicle-stimulating hormone (FSH) and luteinizing hormone (LH)-like activities in rat LHR- and FSHR-expressing cells. In this study, we analyzed signal transduction by eelFSHR and eelLHR upon stimulation with $rec-eCG{\beta}/{\alpha}$ and native eCG. The cyclic adenosine monophosphate (cAMP) stimulation in CHO-K1 cells expressing eelLHR was determined upon exposure to different doses (0-1,450 ng/mL) of $rec-eCG{\beta}/{\alpha}$ and native eCG. The $EC_{50$ values of $rec-eCG{\beta}/{\alpha}$ and native eCG were 172.4 and 786.6 ng/mL, respectively. The activity of $rec-eCG{\beta}/{\alpha}$ was higher than that of native eCG. However, signal transduction in the CHO PathHunter Parental cells expressing eelFSHR was not enhanced by stimulation with both agonist $rec-eCG{\beta}/{\alpha}$ and native eCG. We concluded that $rec-eCG{\beta}/{\alpha}$ and native eCG were completely active in cells expressing eelLHR, similar to the activity in the mammalian cells expressing LHRs. However, $rec-eCG{\beta}/{\alpha}$ and native eCG did not invoke any signaling response in the cells expressing eelFSHR. These results suggest that eCG has a potent activity in cells expressing eelLHR. Thus, we also suggest that $rec-eCG{\beta}/{\alpha}$ can induce eel maturation by administering gonadotropic reagents (LH), such as salmon pituitary extract.

Synthesis of Higenamine and its Cardiovascular Effects in Rabbit: Evidence for ${\beta}-Adrenoceptor$ agonist (Higenamine의 합성 및 가토의 심혈관계에 미치는 영향 : 베타-아드레날린성 효능 약물)

  • Chang, Ki-Churl;Lim, Jung-Kyoo;Park, Chan-Woong
    • The Korean Journal of Pharmacology
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    • v.22 no.2
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    • pp.96-104
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    • 1986
  • Higenamine, dl-1-( 4-hydroxybenzyl)-6, 7-dihydroxy-1 ,2, 3 ,4-tetrahydroisoquinoline has been synthesized and evaluated for hemodynamic actions using rabbits under pentobarbital anesthesia. Concentration-related fall of mean blood pressure was observed, where diastolic blood presure was significantly lowered at 10 ug/kg/min or above (p<.05), while the systolic blood pressure was slightly increased or unaffected, thereby, causing increment of pulse pressure. No significant change was occured in heart rate, however, carotid artery blood flow was significantly (p<.05) increased. These actions were inhibited with pretreatment of 0.3 mg/kg of propranolol, beta-adrenoceptor antagonist, 5 minutes before infusion of higenamine indicating that higenamine compete with propranolol for the so-called beta adrenergic receptor. As comparison, the same procedure was applied to isoproterenol as well, where typical antagonism of propranolol against isoproterenol was shown. From these findings the vasodilating and diastolic blood pressure lowing effects could be explained in terms of cardiac beta stimulating action, however, dopamine receptor activation could not be excluded because no significant changes observed in chronotropism.

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Influence of Intracerebroventricular Clonidine on the Rabbit Renal Function (가토신장기능에 미치는 측뇌실내 Clonidine의 영향)

  • Kook, Young-Johng;Lee, Young-Hoon;Choi, Bong-Kyu
    • The Korean Journal of Pharmacology
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    • v.20 no.2
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    • pp.59-71
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    • 1984
  • To explore the regulatory roles of CNS on the renal function, clonidine, a specific presynaptic ${\alpha}-adrenoceptor$ agonist, was administered into a lateral ventricle of the brain (icv) and the changes of renal function were studies in urethane-anesthetized rabbits. $5{\mu}g/kg$ icv elicited no significant changes in renal function. However, $15{\mu}g/kg$ induced marked natriuresis and kaliuresis for 20 min. Neither RPF nor GFR changed significantly. The fractional sodium reabsorption was significantly reduced, indicating that the renal action was of the tubular origin. Changes of systemic blood pressure were not contributory to the renal action. Yohimbine, a specific antagonist for presynaptic ${\alpha}-adrenoceptor$, when given icv in doses of $100{\mu}g/kg$ 20 min prior to clonidine, completely abolished the renal action of icv clonidine. Yohimbine icv did not produce any significant changes in renal function. Intravenous clonidine, $15{\mu}g/kg$, elicited antidiuresis and decrement of renal function immediately after administration, followed by a slight tendency toward natriuresis, but no natriuresis corresponding to those seen after the icv clonidine were observed, indicating that in the renal action of icv clonidine no direct action is involved. These observations indicate that the central sympathetic tone plays a role in the regulation of renal function in the rabbit.

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