OBJECTIVE: Linkage analysis is a very useful method for prenatal diagnosis of Hemophilia B, especially when a mutation was not identified. Seven polymorphic markers were studied in Korean populations to evaluate the efficiency for prenatal and carrier diagnosis. METHODS: Subjects of this study was 100 healthy Korean women (200 X-chromosomes). Polymerase chain reacton-restriction fragment length polymorphism (PCR-RFLP) method was used to detect SalI, MseI, NruI, DdeI, XmnI, TaqI and HhaI polymorphisms. RESULTS: SalI (-) allele showed the frequency of 0.355 and SalI(+) allele 0.645. MseI(-) allele was 0.645 in frequency and MseI(+) allele was 0.355. SalI and MseI polymorphisms were in complete linkage disequilibrium. And no increase was expected in overall heterozygosity with these two polymorphisms. NruI(-) allele frequency was 0.855 and NruI(+) was 0.145. There was no polymorphism of DdeI, XmnI and TaqI marker systems in Korean population. In HhaI polymorphism, allele frequencies were estimated that HhaI(-) is 0.82 and HhaI(+) is 0.18. CONCLUSION: Only SalI, NruI and HhaI polymorphisms are useful for the diagnosis of hemophilia B in Korean population. Expected heterozygosity for above 3 poylmorphic markers was estimated to be 0.723, and 71 of 100 female subjects were heterozygous for at least one marker system. Korean population showed relatively low extent of polymorphisms compared to Caucasians, Blacks and Japanese. For the effective prenatal diagnosis of hemophilia B with linkage analysis, other polymorphic markers should be evaluated.
African Continental Ancestry Group ; Alleles ; Asian Continental Ancestry Group ; Diagnosis ; Factor IX* ; Female ; Gene Frequency ; Hemophilia B ; Humans ; Linkage Disequilibrium ; Prenatal Diagnosis

No abstract available.
Abortion, Habitual* ; Female ; Immunization* ; Lymphocytes* ; Pregnancy

SUMMARY: The present study was carried out to evaluate whether the coculture system of human embryos with Vero cells can improve the quality of embryo or overcome the repetitive implantation failures in order to obtain pregnancy. From January to December 1996, a total 202 cases which patients with the problems of repetitive implantation failures (group I) or those with the poor embryonic quality in their previous cycles (group II) was analysed. The quality of cocultured embryo, pregnancy, on-going and implantation rates between coculture and control groups were compared. Of 93 cases in group I, coculture was performed in 34 cases and conventional IVF for the rest. Of 109 cases in group II, 36 for coculture and 73 for conventional IVF. In group I, pregnancy, on-going and implantation rates in coculture group (14/34 (41.2%), 9/34 (26.5%), 16/81 (19.8%), respectively) were higher than those of control (11/59 (18.6%), 8/59 (13.6%), 12/152 (7.9%), respectively). There is significance in the pregnancy and implantation rates (p=0.028 and p=0.015). In group II, pregnancy, on-going and implantation rates in coculture group (8/36 (22.2%), 5/36 (13.9%), 8/87 (9.2%), respectively) were higher than those of control (5/73 (6.8%), 3/73 (4.1%), 3/158 (1.9%), respectively). Like the result of group 1, there is significance in the pregnancy and implantation rates (p=0.028 and p=0.022). Coculture system with Vero cells works well in the groups of the two indications. Although the case of 3 day-coculture was small as 15 cases in group II, 3 day-coculture improved pregnancy rate (4/15 (26.7%)). Therefore, 3 day-coculture with assisted hatching is recommended to the patients with poor embryonic quality. In conclusion, coculture system with Vero cells can be suggested as an effective method which improves pregnancy rate in those who have repetitive implantation failures or whose embryonic quality was poor in their previous cycles.
Coculture Techniques* ; Embryonic Structures ; Humans ; Humans* ; Pregnancy ; Pregnancy Rate ; Prognosis* ; Vero Cells

Intracytoplasmic sperm injection (ICSI) recently has been utilized widely as the most successful technique to overcome the unfertilization problem in cases of severe male infertility in couples who could not be treated by conventional IVF. Recently, indications of ICSI have been extended further and more fertilized oocytes become available. Thus, it is necessary to examine the efficiency of freezing the surplus embryos obtained from ICSI. We compared the survival rate and the future outcome of cryopreserved embryos obtained either after conventional IVF or ICSI during the same period. After ICSI or IVF, five best-quality embryos from each patient were transferred in the stimulation cycle and the surplus pronuclear (PN) stage oocytes or multicellular embryos were cryopreserved by slow freezing protocol with 1,2-propanediol (PROH) as a cryoprotectant.4 total of 792 embryos from ICSI trial were thawed and 65.2% (516/792) survived. The survival rates of PN stage oocyte, multicellular embryo and PN + multicellular embryo were 63.5%, 68.2%, 64.0%, respectively. After 111 transfers, 34 pregnancies were achieved, corresponding to a clinical pregnancy rate of 30.6% per transfers. We thawed 1033 embryos from IVF trials and 57.5% (594/1033) survived. In IVF cycle, the survival rates of PN stage oocyte, multicellular embryo and PN + multicellular embryo were 58.2%, 65.2%, 40.2%, respectively. Thirty eight clinical pregnancies were established after 134 transfers, corresponding to a pregnancy rate of 28.4% per transfer The cleavage rate of thawed PN stage oocytes from ICSI trial (61.3%) was significantly higher than those from conventional IVF (53.4%). The developmental rates of good embryo (> or = grade II) in thawed PN stage oocytes obtained from conventional IVF and ICSI were 63% and 65%, respectively. We concluded that PN stage oocytes, multicellular embryos resulting from ICSI procedure can be successfully frozen/thawed with reasonable clinical pregnancy rates comparable to those of IVF.
Embryonic Structures* ; Family Characteristics ; Fertilization in Vitro* ; Freezing ; Humans ; Infertility, Male ; Male ; Oocytes ; Pregnancy ; Pregnancy Rate ; Propylene Glycol ; Sperm Injections, Intracytoplasmic* ; Survival Rate

This study was carried to determine the possibility of finding motile spermatozoa and fertilization, pregnancy rate after testicular sperm extraction(TESE) with ICSI in obstructive and non-obstructive azoospermic patients. In 154 cases(132 patients), obstructive azoospermia was 77 cases and non-obstructive azoospermia was 77 cases. In obstructive azoospermia, patients generally showed normal spermatogenesis and included vas agenesis(n=8), multiple vas obstruction(n=7), epididymal obstruction (n=54). Total of 982 retrieved oocytes were obtained and 84.4% were injected. The fertilization rates with 2 PN and cleavage rate were 72.5% and 62.3%, .respectively. 30 pregnancies(38.9%) were achieved and the ongoing pregnancies were 22 cases (28.6%). In non-obstructive azoospermia, patients showed hypospermatogenesis(n=49), maturation arrest(n=4), Sertoli cell only syndrome (n=24). The various stages of spermatogenic cell could be retrieved by TESE and could be reached normal fertilization and embryo development with ICSI. Total of 1072 retrieved oocytes obtained and 80.2% were injected. The fertilization rates with 2 PN and cleavage rate were 52.8% and 68.9%, respectively. 22 pregnancies(30.1%) were achieved and the ongoing pregnancies were 19 cases(26.0%). Conclusively, the combination of TESE with ICSI using testicular spermatozoa can achieve normal fertilization and pregnancy rate and effective method in obstructive and non-obstructive azoospermic patients.
Azoospermia ; Embryonic Development ; Female ; Fertilization* ; Humans ; Oocytes ; Pregnancy Rate* ; Pregnancy* ; Sertoli Cell-Only Syndrome ; Sperm Injections, Intracytoplasmic ; Spermatogenesis ; Spermatozoa*

Irreparable obstructive azoospermic patients can be treated successfully with microsurgical epididymal sperm aspiration(MESA) o. testicular sperm extraction (TESE) by intracytoplasmic sperm injection(ICSI). Obstructive azoospermic patients generally have normal spermatogenesis. The aim of this study was to see if any spermatozoa could be retrieved from non-obstructive azoospermia and to assess the efficacy of ICSI with TESE in germinal failure. 42 non-obstructive azoospermic patients revealed no spermatozoa at all in their ejaculates, even after centrifuge. The histology of 42 patients revealed 15 Sertoli cell only Syndrome, 4 maturation arrest and 23 severe hypospermatogenesis. All patients underwent extensive multiple testicular biopsy for sperm retrieval. These patients were scheduled for ICSI using testicular spermatozoa. In 25 out of 42 non-obstructive azoospermic patients, spermatozoa were recovered from multiple testicular biopsy specimen and 11 ongoing pregnancies were achieved. There are usually some tiny foci of spermatogenesis which allow TESE with ICSI in non-obstructive azoospermia. Also these patients may have sufficient sperm in the testes for ICSI, despite extremely high FSH level and small testes.
Azoospermia* ; Biopsy ; Humans ; Male ; Oligospermia ; Pregnancy ; Sertoli Cell-Only Syndrome ; Sperm Injections, Intracytoplasmic ; Sperm Retrieval ; Spermatogenesis ; Spermatozoa ; Testis

OBJECTIVES: The aims of this study are 1) to determine if GAST is a better indicator in predicting ovarian response to COH compared with patient's age or basal FSH level and 2) to evaluate its role in detecting abnormal ovarian response. DESIGN : Prospective study in 118 patients undergoing IVF-ET using GnRH-a short protocol during May-September 1995. MATERIALS AND METHODS: After blood sampling for basal FSH and estradiol (E2) on cycle day two, 0.5 ml (0.525 mg) GnRH agonist (Suprefact, Hoechst) was injected subcutaneously. Serum E2 was measured 24 hours later. Initial E2 difference (deltaE2) was defined as the change in E2 on day 3 over the baseline day 2 value. Sixteen patients with ovarian cyst or single ovary or incorrect blood collection time were excluded from the analysis. The patients were divided into three groups by deltaE2; group A (n=30):deltaE2<40 pg/ml, group B (n=52): 40 pg/ml< or =deltaE2<100 pg/ml, group C (n=20): deltaE2< or =100 pg/ml. COH was done by GnRH agonist/HMG/hCG and IVF-EF was followed. Ratio of E2 on day of hCG injection over the number of ampules of gonadotropins used (E2hCGday/Amp) was regarded as ovarian responsiveness. Poor ovarian response and overstimulation were defined as E2 hCGday less than 600 pg/ml and greater than 5000 pg/ml, respectively. RESULTS: Mean age (+/-SEM) in group A, B and C were 33.7+/-0.8*, 31.5 +/-0.6 and 30.6+/-0.5*, respectively (*: p<0.05). Mean basal FSH level of group A (11.1+/-1.1 mIU/ml) was significantly higher than those of B (7.4+/- 0.2 mIU/ml) and C (6.8+/-0.4 mIU/ml) 0<0.001). Mean E2hCGday of group A was significantly lower than those of group B or C, i.e., 1402.1+/-187.7 pg/ml, 3153.2+/- 240.0 pg/ml, 4078.8+/-306.4 pg/ml respectively (p<0.0001). The number of ampules of gonadotropins used in group A was significantly greater than those in group B or C: 38.6+/-2.3, 24.2+/-1.1 and 18.5+/-1.0 (p<0.0001). The number of oocytes retrieved in group A was significantly smaller than those in group B or C: 6.4+/-1.1, 15.5+/-1.1 and 18.6+/-1.6, respectively (p<0.0001). By stepwise multiple regression, only deltaE2 showed a significant correlation (r=0.68, p<0.0001) with E2HCGday/Amp, while age or basal FSH level were not significant. Likewise, only deltaE2 correlated significantly with the number of oocytes retrieved (r=0.57, p<0.001). All four patients whose COH was canceled due to poor ovarian response belonged to group A only (Fisher's exact test, p<0.01). Whereas none of 30 patients in group A (0%) had overstimulation, 14 patients among 72 patients (19.4%) in group B and C had overstimulation (Fisher's exact test, p<0.01). CONCLUSIONS: These data suggest that initial E2 difference after GAST may be a better prognostic indicator of ovarian response to COH than age or basal FSH level. Since initial E2 difference demonstrates significant association with abnormal ovarian response such as poor ovarian response necessitating cycle cancellation or overstimulation, GAST may be helpful in monitoring and consultation of patients during COH in IVF-ET cycle.
Estradiol ; Female ; Gonadotropin-Releasing Hormone* ; Gonadotropins ; Humans ; Oocytes ; Ovarian Cysts ; Ovary ; Ovulation Induction* ; Prospective Studies

Uterine anomalies have been reported in 4% of women with infertility and in up to 15% of those with recurrent abortion. One of the major intrauterine disorder associated with infertility and recurrent abortions is intrauterine septum, The reproductive outcome of 41 patients of intrauterine septum (7 complete, 34 incomplete) with repeated abortions or infertility was assessed after the uterine septotomy. 5 of 7 patients with comlete uterine septum undergone uterine septotomy (3; hysteroscopic metroplasty, 2; abdominal metroplasty) had total 6 pregnancies and all of them had live biths. 28 patients with incomplete uterine septum got the hysteroscopic intrauterine septotomy and the viable pregnancy rate was 62% (3 ongoing pregnancies, 13 live biths of total 26 pregnancies). 6 patients with incomplete uterine septum had not the operation and 5 patients had 5 live births after total 6 pregnancies with 1 spontaneus abortion. Even though, the number of cases were small, the live birth rate in the group of septotomy of the patients of complete uterine septum (100%, 6/6) was higher than that in the group of not-done (50%, 1/2). The live birth rate in the group of not-done of the patients with incomplete uterine septum (83%, 5/6) was higher than that in the group of hysteroscopic uterine septotomy (62%, 16/26), but 5 of 6 had short uterine septal length (<1 cm), 1 had 1.5 cm septal length in the group of not-done. All the patients with successful pregnancy outcome had no other co-factors at the diagnostic laparoscopy, but the 5 primary infertility patients with no live birth even after treatment (all were with incomplete septum; 3 undergone hysteroscopic septotomy, 2 not-done with one abortion) had other co-factors such as endometriosis, peritoneal or tubal facor. In conclusion, hysteroscopic uterine septotomy would be useful for the patients with habitutal abortion or infertility and more advanced managemnet protocols should be applied to the patients having other co-factors if there was no pregnancy even after the uterine septotomy.
Abortion, Habitual* ; Endometriosis ; Female ; Humans ; Hysteroscopy* ; Infertility* ; Laparoscopy ; Live Birth ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate

In our previous study, we observed that hydrosalpingeal fluid (HSF) adversely effect mouswe embryo development and hatching. The aim of this study was to evaluate the effect of HSF as assessed by the blastocyst development rate (BDR) and by cell counting in vitro HSF was collected from nine patients undergoing salpingoneostomy to correct hydrosalpinx. Two-cell embryos were obtained from superovulated ICR mice. T6 medium and T6+/-0.4% bovine serum albumin were used as control media. T6 medium containing 10% or 50% HSF and 100% HSF from each patient were used as test media. Nine to 15 embryos were cultured in microdrops prepared from each of these media. To assess the total cell number within each blastocyst, the blastocysts were fixed and stained with Hoechst 33342 to facilitate cell counting. The mean BDR in two control media were 88.89% and 85.40%. The mean BDR in media containing 10%, 50%, 100% HSF were 85.87%, 89.58% and 75.57%, respectively (*: p<0.05). The overall mean cell count (+/-SEM) in control media were 87.6+/-9.65 and 90.12+/-11.38. The BDR was affected adversely only by 100% HSF and not in media containing 10% or 50% HSF. Mean cell counts were decreased significantly only in blastocysts cultured 100% HSF (63.8+/-13.66; p<0.01) but not in blastocysts cultured in 10% or 50% HSF (91.3+/-12.44 and 82.9+/-18.27, respectively). Thus, it is concluded that HSF has no embyotoxic effect but has a mildly negatively effect on embryonic growth and development.
Animals ; Blastocyst ; Cell Count ; Embryonic Development ; Embryonic Structures* ; Female ; Growth and Development ; Humans* ; Mice* ; Mice, Inbred ICR ; Pregnancy ; Serum Albumin, Bovine

BACKGROUND: Antihypertensive agents such as verapamil and esmolol are well known for their effects of hemodynamic stabilization on tracheal intubation. However, our previous study, Verapamil and esmolol did not attenuate heart rate and blood pressure. The aim of the present study was to evaluate the efficacy of combined administration of these drugs for controlling hemodynamic responses to tracheal intubation. METHODS: Forty-eight patients, ASA physical status I or II, were randomly assigned to one of four groups (n = 12 each):normal saline (control), verapamil 0.1 mg/kg, esmolol 1 mg/kg, and verapamil 0.05 mg/kg mixed with esmolol 0.5 mg/kg. Anesthesia was induced with thiopental 5 mg/kg intravenously, and then saline, verapamil, esmolol or the mixed drugs were administered as an intravenous bolus, and immediately followed by succinylcholine 1.5 mg/kg. Tracheal intubation was performed 90 s after intravenous injection of experimental drugs. Systolic and diastolic blood pressure and heart rate were measured before induction and every minute for 5 minutes after tracheal intubation. RESULTS: There was a significant attenuation in systolic blood pressure after tracheal intubation in the verapamil and mixed groups compared to the control and esmolol groups. Heart rates were significantly lower in the esmolol and mixed groups than in the verapamil groups after tracheal intubation. CONCLUSIONS: Combined administration of Verapamil 0.05 mg/kg with esmolol 0.5 mg/kg attenuated increases in blood pressure and heart rate after tracheal intubation.
Anesthesia ; Antihypertensive Agents ; Blood Pressure* ; Heart Rate* ; Heart* ; Hemodynamics ; Humans ; Injections, Intravenous ; Intubation* ; Succinylcholine ; Thiopental ; Verapamil*