• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.9-14
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• 2000

Objective: The sperm acrosome reaction is a $Ca^{2+}$-dependent exocytotic event that is triggered by adhesion to the mammalian egg's zona pellucida. Previous studies suggested a role of $Ca^{2+}$ channels in acrosome reactions. This study was conducted to investigate the T-type calcium channel is operated in acrosome reaction of human spermatozoa. Method: Human semen samples were obtained from healthy donors with normal criteria. The spermatozoa were divided into five groups: Group 1 were non-treated as a control; Group 2 where spermatozoa were exposed to 5 ${\mu}M$ $Ca^{2+}$ A23187 $(Ca^{2+}i)$; Group 3 where spermatozoa were exposed 5 ${\mu}M$ $Ca^{2+}i$ and mibefradil; Group 4 where spermatozoa were exposed 5 ${\mu}M$ $Ca^{2+}i$ and nifedipine, and Group 5 where spermatozoa were treated with 5 ${\mu}M$ $Ca^{2+}i$ and both of mibefradil and nifedipine. Spermatozoa in all groups were retrieved after incubation for 15 and 30 minutes at $37^{\circ}C$. After staining with PSA-FITC, fluorescence was observed under a fluorescence microscope, and AR was evaluated on a total>100 spermatozoa/side. Result and Conclusion: We observed on acrosome reaction inhibition rate in human spermatozoa the various of concentration of mibefradil, nifedipine. Maximum response was noted with 1.0 ${\mu}M$ mibefradil and the decrease of acrosome reaction inhibition rate 45%. Nifedipine in acrosome reaction inhibition rate was only about 25%. The $Ca^{2+}i$-induced AR of spermatozoa was significantly suppressed by mibefradil. Incidence of the suppression was depending on concentration of mibefradil. Results from the present study suggest that the human spermatozoa possess T-type channel. The observation that reversible inhibitor of T channels in male germ cells provides a new mechanism of contraceptive action.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.2
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• pp.165-172
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• 2000

Objective: In vitro fertilization (IVF) and a prolonging the time of culture may be helpful in establishing a viable pregnancy through a selection effect. Some embryos do not develop beyond the 4-cell stage and some may not develop to the blastocyst stage. We have evaluated the safety of SET and the outcomes of pregnancy. Methods: Sperms were treated with Ham's F-10 supplemented with 10% human follicular fluid (hFF). oocytes or fertilized oocytes were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% or 20% hFF respectively. Up to five oocytes were inseminated with approximately 200,000 sperm cells/2 ml in each well. Fertilization was examined in the following morning and fertilized oocytes were co-cultured until embryo transfer. Vero cells for co-culture were prepared in Tissue Culture Medium - 199 (TCM-199) with 10% fetal bovine serum. At the two to four cell and blastocyst on day 2 and day 5, embryo and blstocyst grading were evaluated. Pregnancy rate was determined after transfer of human embryos at the two to four cell stage on day 2 (Group I) or subsequent transfer of embryos on day 2 and at the blastocyst stage on day 5 (Group II). For statistical analysis, Student's t-test and Chi-square (${\chi}^2$_test) were used. Results were considered statistically significant when p value was less than 0.05. Results: No differences was found in the fertilization between Group I (81.0%, 98/121) and Group II (81.8%, 180/220). In case of cleavage rate, no difference was found in Group I (95.9%, 94/98) and Group II (97.8%, 174/178). However, the rate of-clinical pregnancy was significantly higher (p=0.014) in Group II (66.7%, 12/18) than in Group I (26.3%, 5/19). Conclusion: The results of this study showed that SET is safe and effective, and significantly increases the pregnancy rate.

• Research in Community and Public Health Nursing
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• v.7 no.1
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• pp.138-153
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• 1996

Intending to offer basic information for a prospective health services in Korea, this study is to investigate the complication caused by sterilization in goverment family planning program from 1962 to 1995. The results are as follows: 1. Total number of sterilization performed during the period from 1962 to 1995 were 1.367,772 cases of male sterilization and 2,889,635 cases of female sterilization. 2. Incidence of the complication caused by sterilization operation from 1980 to 1995 were 1,883(0.20%) out of 925,801 cases in vasectomies and 15,866(0.70%) out of 2,256,020 cases in tubal sterilizations. 3. Major complications in vasectomy were epididymities of 658 cases (34.9%), vas recanalization of 326 cases(17.3%), hematoma of 266 cases(14.1%), scrotal abscess of 184 cases(9.8%), sperm granuloma of 76 cases(4.0%),and other of 373 cases(19.8%). On the other hand, in tubal sterilization, ectopic pregnancy was the most significant complication of 15,078 cases (95.0%) among 15,866 total complications, followed by pelvic inflammatory diseases of 155 cases(0.9%), peritonities of 96 cases(0.6%), ovarian & tubal bleeding of 31 cases(0.2%), intestinal perforation of 16 cases (0.1%), uterine bleeding of 14 cases(0.1%), uterine cervix laceration of 1 case (0.1%), and other of 271 cases(1.7%), while 161 pregnancies(0.1%) were terminated and 43 cases(0.3%) with normal delivery. 4. The occurrence rate of the complication for each period showed that most of the complication cases by vasectomy occurred in a year after the operation -the cases were 1,256 (66.7%). 254 cases(13.5%) occurred between the next year and the 2nd year, 138 cases (7.3%) between the 2nd year and the 3rd year, 73 cases(3.9%) between the 3rd year and the 4th year, 52 cases(2.8%) between the 4th year and the 5th year, 31 cases(1.6%) between the 5th year and the 6th year, 79 cases(4.2%) over the 6th year. Tubal sterilization indicated that the occurred complication cases in a year were 2,175 cases(13.7%), 2,113 cases(13.3%) occurred between the next year and the 2nd year, 2,082 cases(13.1%) between the 2nd year and the 3rd year, 2,049 cases (12. 9%) between the 3rd year and the 4th year, 1,819 cases(11.5%) between the 4th year and the 5th year, 621 cases(10.2%) between the 5th year and the 6th year, 4,007 cases(25.3%) over the 6th year. 5. For the cost of complication treatment, total \7,928,229,000 were paid as medical expenditure in which \609,438,000 for vasectomy and \7,318,791,000 for tubal sterilization. Accordingly per capita expenses were \345,000 for vasectomy and \467,000 for tubal sterilization. As the proportion of government sterilization program was decreased after 1988, that of private sterilization program would be increased. So it is recommended to set a guideline for the private sterilization program and to continue government sterilization program for the lower class.

• Journal of Aquaculture
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• v.10 no.1
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• pp.25-32
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• 1997

Develoment precess and characteristics of eggs of the geoduck clam, Panope japonica are reporting in this study. Eggs and sperm were excised from gonad, artificially fertilized in an aquarium, reared under various temperature regimes, and record and record the larval period and the time need to reach a certain larval stage from ferilization. Unfertilized eggs of P. japonica appeared to be oval with a mean diameter of $70\mu$m and they became spherical after fertilization. The eggs of P. japonica can be classified as demersal. At a constant water temperature of $ 11^{citc}C$, it took 4 hours form fertilization to become four-cell stage, two days to become trochophore larvae, three days to become D-shape larvae, twenty-three days to become umbo stage, and thirty-six days to become fully grown veliger ready form settlement. A negative correlation was observed between the water temperature and the larval period of P. japonica. From fertilization to D-shape larvae, it took five days at 8$^{\circ}C$, while it was only two days to become D-shape larvae at $ 17^{citc}C$. Time required to D-shape larvae from fertilization was proportional to temperature, and the relationships were expressed as follows : To 8-cell stage, 1/t=0.0209 w-0.1167 (r=0.9967) To blastula stage, 1/t=0, 0055 w-0.0192 (r=0.9825) To trochophore stage, 1/t=0.0034 w-0.0155 (r=0.9907) To D-shape larvae stage, 1/t=0.0014 w-0.0023 (r=0.9843) (t, time in hours ; w, water temperature) Bioligical minimum temperature for egg development was calculated as 3.82$^{\circ}C$ in average.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.3
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• pp.339-344
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• 1999

Objective: To evaluate incidence of microbial growth from the tip of the embryo transfer catheter after embryo transfer in relation to clinical pregnancy rate following in-vitro fertilization and embryo transfer. Method: This study was performed prospectively at the time of transcervical embryo transfer following conventional in-vitro fertilization and intracytoplasmic sperm injection procedures. Sixty three patients were enrolled in this study. Microbiological cultures were performed on endocervical swabs and embryo transfer catheter tips. Results: Positive microbial growths were observed from endocervical swabs in 45 (71.4%) women and from catheter tips in 30 (47.6%) women. There was no statistically significant difference seen in the mean number of oocytes fertilized or number and grade of embryos transferred between the group of patients without growth and the group of patients with positive microbial growth from catheter tips. The clinical pregnancy rate were 30.3% in the group of patients without growth and 13.3% in the group with positive microbial growth from catheter tips. This difference in clinical pregnancy rates was statistically significant. Conclusion: Our finding is that microbial contamination at embryo transfer may influence implantation rates. The major questions arising from our finding are whether eradication of endocervical micro-organisms is possible and whether their eradication will improve implantation rates.

• Korean Journal of Ichthyology
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• v.29 no.2
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• pp.101-108
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• 2017

Egg development and early life history of the Korean natural monument fish Hemibarbus mylodon (Pisces: Cyprinidae) were investigated to provide basic data on biological characteristics and ecological recovery. Adult fish were collected from nature and transferred to the laboratory. For the first time, artificial maturation of females and males succeeded after 15 months of indoor culture. Mature eggs and sperm were obtained using Ovaprim injections (0.5 mL/kg) and then the eggs were fertilized using the dry method in the laboratory. The mature eggs were adhesive, turbid, and greyish; they averaged $2.21{\pm}0.06mm$ (n=30) in diameter. The embryos began to hatch about 78 h after fertilization at a water temperature $20{\pm}1^{\circ}C$, and the newly-hatched larvae were $6.6{\pm}0.75mm$ in total length (TL). At 14 days after hatching, the post-larval individuals were $13.5{\pm}0.23mm$ (TL), and their yolk sacs were completely absorbed. Twenty one days after hatching, they entered the juvenile stage and reached $13.5{\pm}0.23mm$ (TL). At 100 days after hatching, their band patterns, external form, and a pair of barbels were similar to those of adults, and they averaged $33.0{\pm}4.25mm$ (TL). The breeding technology and characteristics of early life history obtained in this study will be very helpful for conservation of H. mylodon in nature.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.4
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• pp.333-340
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• 2003

Objective: This study was performed to examine the maturation and the development to the blastocyst stage of immature oocytes collected from patients with high risk of ovarian hyperstimulation syndrome (OHSS). Materials and Methods: Cumulus-oocyte complexes (COCs) were collected following only HCGpriming for non stimulated IVF-ET cycles of the patients. At the time of oocyte collection, COCs were classified into three groups in accordance with their appearance (Group I: oocytes with dispersed cumulus cells; Group II: oocytes with compacted cumulus cells; Group III: oocytes with sparse cumulus cells). The in vitro maturation and blastocyst development rates of the COCs were compared among these groups. From August 2001 to June 2002, 48 IVM/IVF-ET cycles from 42 patients (mean age: $32.4{\pm}3.8$ years) were performed. To prevent the occurrence of OHSS, the patients were primed with 10, 000 IU HCG alone 36 h before oocyte collection without gonadotropin stimulation. Oocytes were aspirated on cycle days from 7 to 13. The normal COCs were classified into three groups according to their appearance. The aspirated immature oocytes were cultured in YS maturation medium containing 30% (v/v) human follicular fluid (HFF), 1 IU/ml FSH, 10 IU/ml HCG and 10 ng/ml rhEGF. Fertilization was induced by intracytoplasmic sperm injection (ICSI). All zygotes were co-cultured with cumulus cells in $10{\mu}l$ YS medium containing 10% HFF until day 7 after oocyte collection. Blastocyst transfer was performed on day 5 after ICSI. Results: Th e mean number of oocytes cultured in the IVM/IVF cycles was $24.7{\pm}10.6$. Of 1185 COCs, those assigned to Group I, II and III were 470 (39.7%), 414 (35.0%) and 301 (25.4%), respectively. The maturation rate (94.5%, 444/470, p<0.05) in Group I was significantly higher than those of Group II (62.8%, 260/414) and Group III (73.1%, 220/301). Especially, 30.9% of COCs in Group I (145/470) was matured on the day of oocyte aspiration. There were no differences in the rates of fertilization and cleavage among the three groups. The development rate to the blastocyst stage in Group I (54.6%, 206/377, p<0.05) was also significantly higher than those in Group II (33.0%, 68/206) and Group III (30.1%, 52/173). Twenty-four clinical pregnancies (50.0%) was obtained and 22 pregnancies (45.8%) are ongoing. Implantation rate in the present study was 24.6%. Conclusion: These results suggest that there is a positive correlation between the appearance of COCs and the developmental competence of the immature oocytes in non stimulated IVM/IVF cycles.

• 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.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.1
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• pp.31-36
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• 2011

Objective: To determine whether the serum ${\beta}$-human chorionic gonadotropin (hCG) profile following preimplantation genetic diagnosis (PGD) is lower than that of intracytoplasmic sperm injection (ICSI) cycles. Methods: A total of 129 PGD cycles and 1,161 age-matched ICSI cycles, which resulted in pregnancy (serum ${\beta}-hCG{\geq}5$ mIU/mL) on post-ovulation day (POD) 12 were included. We compared the mean serum ${\beta}$-hCG levels on POD 12, 14, 21, and 28, doubling time of serum hCG, and created a cut-off value for predicting a singleton pregnancy in each group. Results: The mean serum ${\beta}$-hCG concentration of the PGD group was significantly lower than that of the control group on POD 12, 14, and 21. The doubling time of serum ${\beta}$-hCG at each time interval showed no significant difference. The cut-off-value of serum ${\beta}$-hCG for predicting a single viable pregnancy was 32.5 mIU/mL on POD 12 and 113.5 mIU/mL on POD 14 for the PGD group, which was lower than that for the control group. Conclusion: Blastomere biopsy may decrease the ${\beta}$-hCG-producing activity of the trophoblasts, especially in early pregnancy. Setting a lower cut-off value of serum ${\beta}$-hCG for predicting pregnancy outcomes in PGD may be needed.

• Korean Journal of Medicinal Crop Science
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• v.2 no.1
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• pp.81-85
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• 1994

This experiment was carried out to investigate the fertilization and the size of mature female and male gametophytic parts of Pulsatilla koreana after artificial pollination. The size of pollen is $26.5{\mu}m$ at the time of anther dehiscence, that is, about $3{\sim}4$ days after pollination. Synergid nucleus, egg nucleus, and polar nucleus are 10.0, 15.0, and $32.5{\mu}m$ respectively at the time of completing egg apparatus formation, that is, about 2 days after pollination. Poller tubes germinate on stigma about 10 hours, passing lower part of style about 30 hours, penetrating into micropyle about 35 hours after pollination. Sperm nucleus penetrates into polar nucleus about 40 hours and egg cell about 48 hours after pollination. But, there seems to be different among the individuals. Multinuclei and multinucleoli are formed in egg cell, synergid, and polar nucleus about the time of fertilization. Proembryo is formed about 4 days, being changed to large globular form about $6{\sim}8$ days after pollination. Endosperm nuclei divide into free nuclei after fertilization and change to cotyledon in gymnosperm. There seems to be same phenomena in Pulsatilla koreana.