Does Post-Warming Extended Culture Duration Affect the Clinical and Obstetric Outcomes of Patients of Advanced Maternal Age? A SingleCenter Study

Background@#The single vitrified-warmed blastocyst transfer (SVBT) cycle has been increasingly utilized for assisted reproductive technology. Women of advanced maternal age (AMA) comprise a significant portion of patients who have undergone ‘freeze-all’ cycles. This study investigated the association between the post-warming extended culture duration and pregnancy outcomes in patients of AMA. @*Methods@#This retrospective cohort study analyzed the outcomes of 697 SVBT cycles between January 2016 and December 2017. The cycles were divided into 3 groups based on the age of the female partners: group I: < 35 years (n = 407), group II: 35–37 years (n = 176); and group III, 38–40 years (n = 114). Data are shown as the mean ± standard error of the mean. Data were analyzed using one-way ANOVA followed by Duncan’s multiple range test. Statistical significance was set at P < 0.001. @*Results@#The blastocyst rate, clinical pregnancy rate, and live birth rate (LBR) was significantly lower in the AMA groups. However, there were no significant differences in LBR in the transfer between the AMA and younger groups according to blastocyst morphology and post-warming extended culture duration. @*Conclusion@#Post-warming extended culture of blastocysts is not harmful to patients of AMA. It could be a useful parameter in clinical counseling and decision making for fertility treatments.

Correlation between Morphologic Grading and Euploidy Rates of Blastocysts, and Clinical Outcomes in In Vitro Fertilization Preimplantation Genetic Screening

BACKGROUND: The standard morphological evaluation has been widely used for embryo selection, but it has limitations. This study aimed to investigate the correlation between morphologic grading and euploidy rate of in vitro fertilization (IVF) preimplantation genetic screening (PGS) and compare the pregnancy rates in young and old ages. METHODS: This is a retrospective study using the medical records of patients who underwent IVF procedures with PGS between January 2016 and February 2017 in a single center. The embryo grades were categorized into 4 groups: excellent, good, fair, and poor. Basic characteristics, euploidy rates, clinical pregnancy (CP) rates and ongoing pregnancy rates were analyzed. RESULTS: The excellent group had significantly higher rate of euploid embryos than fair group (47.82% vs. 29.33%; P = 0.023) and poor group (47.82% vs. 29.60%; P = 0.005). When the four groups were recategorized into two groups (excellent and good vs. fair and poor), they also showed significant difference in euploidy rates (44.52% vs. 29.53%; P = 0.002). When the patients were divided into two groups by age 35, the CP rates for those under and over 35 years old were 44.74% and 47.83%, respectively, which showed no significant difference. CONCLUSION: The significant differences among the euploidy rates of different morphologic embryo grades demonstrated the positive correlations between the morphologic grading of the embryo and the euploidy rate of PGS. Additionally, there was no significant difference between the younger and older patients' CP rates. These findings emphasize the fact that old age patients might benefit from PGS whatever the indication of PGS is.

Management of Recurrent Pregnancy Loss

Recurrent pregnancy loss (RPL) is defined as three or more consecutive spontaneous abortions. It affects about 1% of couples attempting pregnancy. Most of these pregnancy losses are unrecognized. Among the pregnancies that are clinically recognized, loss occurs in 15% before 20 weeks of gestation. Only about 50% of women with RPL have an identifiable etiology. Genetic abnormalities are responsible for RPL in 2~4% of these couples, most with balanced translocation. Luteal phase defect might be present in 25~30% of women with RPL; however, progesterone supplementation for miscarriage prevention has not been adequately studied. Uterine abnormalities might also result in RPL. Inherited thrombophilias appear to be associated with fetal death, and antenatal thromboprophylaxis might be beneficial for some women with an inherited thrombophilia. Antiphospholipid syndrome is known to cause RPL, and antenatal thromboprophylaxis reduces the risk of miscarriage. The evidence on alloimmune incompatibility as a cause of RPL is preliminary and no immunotherapy has been shown to be effective in the prevention of miscarriage. Recently a series of new factors that exhibit an association with RPL have been identified, and they might exhibit an additive or multiplicative effect on the RPL risk.

An analysis of various factors affecting postpartum umbilical cord blood volume for hematopoietic stem cell transplantation / 대한산부인과학회지

OBJECTIVE: Umbilical cord blood is an effective alternative to bone marrow as a source of hematopoietic stem cells for transplantation. But the amount of collected umbilical cord blood and its contents are limited and obtaining an adequate volume of umbilical cord blood is essential for successful transplantation. The aim of this study was to identify factors that influence the volume of umbilical cord blood. METHODS: A retrospective analysis of the maternal, neonatal and placental factors that were obtained by medical record review was conducted. The variables that were evaluated for this study were mother's age, parity, gestational age, presence of maternal diabetes mellitus, route of delivery, multiple births, neonatal sex and birth weight, and placental weight. Total 484 deliveries were evaluated from March 2003 to April 2004. The statistical significance of observed differences was calculated using t-test and multiple regression analysis; p-value<0.05 was considered significant. RESULTS: Gestational age, neonatal birth weight, placental weight, parity, number of fetus and maternal diabetes mellitus were significantly associated with a greater volume of collected umbilical cord blood. Obstetric factors that influenced the total nucleated cell concentration were gestational age, neonatal birth weight, placental weight, number of fetus, and route of delivery. CONCLUSION: To prolong a gestational age as far as possible, at least beyond the 37 completed weeks of gestation, and modifying a method of vaginal delivery or cesarean section rather than conventional vaginal delivery method can increase significantly the volume of collected cord blood and the yield of the concentration of total nucleated cell.

Clinical efficacy of GnRH antagonist multiple dose protocol and GnRH agonist long protocol in controlled ovarian hyperstimulation using rFSH in IVF-ET patients / 대한산부인과학회지

OBJECTIVE: To compare the efficacy of GnRH antagonist multi dose protocol in controlled ovarian hyperstimulation (COH) for IVF-ET or ICSI with GnRH agonist long protocol. METHODS: From January 2003 to December 2004, total of 583 cycles which underwent IVF-ET or ICSI using r-FSH were enrolled in this study. 447 cycles of the study group were performed in controlled ovarian hyperstimulation by using GnRH antagonist multi dose protocol and 136 cycles of the control group were performed by using GnRH long protocol. We compared patients characteristics, controlled ovarian hyperstimulation outcomes and IVF-ET outcomes between two groups. RESULTS: Patients characteristics and baseline hormone levels were not different between the two groups. The duration of stimulation was significantly shorter in study group comparing with control group (12.8+/-1.5 days vs 13.7+/-1.7 days, p<0.05). There were no differences between the two groups in the number of follicles, endometrial thickness and serum E2 level on hCG day. The pregnancy rate seemed to be lower in the study group (32.4% vs 35.4%), but the difference was not statistically significant. There were also no differences in number of oocytes retrieved, matured oocytes, fertilized oocytes and transferred embryos between two groups. CONCLUSION: GnRH antagonist multi dose protocol in COH might be a simple and effective method compared with GnRH agonist long protocol.

Accumulation of mtDNA Deletion (Delta mtDNA4977) showing Tissue-Specific and Age-Related Variation / 대한불임학회지

OBJECTIVES: Controversial arguments exists on both the case for and against on the accumulation of mitochondrial DNA (mtDNA) deletion in association to tissue and age. The debate continues as to whether this mutation is a major contributor to the phenotypic expression of aging and common degenerative diseases or simply a clinical insignificant epiphenomenon. The objective of this study was to determine whether the accumulation of mtDNA deletion is correlated with age-related and tissue-specific variation. MATERIALS AND METHODS: One hundred and fifty-seven tissues from blood, ovary, uterine muscle, and abdominal muscle were obtained from patients ranging in age from 31~60 years. After reviewing the clinical reports, patients with mitochondrial disorder were excluded from this study. The tissues were obtained at gynecological surgeries with the consent of the patient. Total DNA isolated from blood, ovary, uterine muscle, and abdominal muscle was amplified by two rounds of PCR using two pairs of primers corresponding to positions 8225-8247 (sense), 13551-13574 (antisense) for the area around deleted mtDNA and 8421-8440 (sense), 13520-13501 (antisense) for nested PCR product. A statistical analysis was performed by c2-test. RESULTS: About 0% of blood, 94.8% of ovary, 71.4% of uterine muscle, and 86.1% abdominal muscle harbored mtDNA deletion. When we examined the proportion of deleted mtDNA according to age deletion rate was 90% of ovary, 63.6% of uterine muscle, 77.7% of abdominal muscle in thirties and 100% of all tissue in fifties. CONCLUSION: The findings of this study suggest that the mtDNA deletion is varied in tissue-specific pattern and increases with aging.

The Study of 5,10-Methylenetetrahydrofolate Reductase Variation (MTHFR C677T) in Infertile Females with Polycystic Ovarian Syndrome (PCOS) in Korea / 대한불임학회지

OBJECTIVE: To investigate the association of genetic background between MTHFR C677T genotype and infertile females with polycystic ovarian syndrome. MATERIALS AND METHODS: We compared 86 infertile females with polycystic ovarian syndrome (PCOS) with 100 healthy fertile females with one or more offspring. Pyrosequencing analysis for MTHFR C677T variation was performed on polymerase chain reaction (PCR) product of study group. To validate pyrosequencing data of C677T variation for randomly selected 50 samples, we compared the pyrosequencing result with the PCR-RFLP (Restriction Fragment Length Polymorphism) result of MTHFR C677T genotype. RESULTS: The prevalence of the C677T mutant homozygous (TT) was significantly lower (p=0.0085) in females with PCOS (8.14%) than in fertile females (21.00%). MTHFR 677 TT genotype had a decreased risk (3.7-fold) of PCOS compared with wild type (MTHFR 677 CC). CONCLUSION: Our data support a role for MTHFR mutant homozygous (677 TT) genotype in reducing risk in Korean infertile females with Polycystic ovarian syndrome.

The Analysis of SHP (Small Heterodimer Partner) Gene Mutation in Infertile Patients with Polycystic Ovary Syndrome (PCOS) in Korea / 대한불임학회지

OBJECTIVE: We inversigated Small Heterodimer Partner (SHP) gene mutation in Korean Polycystic Ovarian Syndrome (PCOS) patients. SHP protein regulates the activity of nuclear receptors which regulate the cellular development and differentiation. Recently, the mutation of SHP gene was found in the obesity and diabetes patients in Japanese group, and suggested that its mutation may involved in pathogenic mechanism of PCOS. METHODS: This study was performed in 20 PCOS patients and 20 normal women. The DNAs were extracted from the peripheral bloods, and amplified at each exon (1 and 2) of SHP gene by PCR method. Subsequently, each PCR product was digested with the restriction enzyme indicated below for studying restriction fragment length polymorphism (RFLP). After enzyme digestion, the results of RFLP were compared PCOS patients with control women to find any sequence variation. RESULTS: We examined 9 regions of exon 1 with Msp I, Pvu II, Dde I and 3 regions of exon 2 with Pst I, Dde I. There is no heterozygous or homozygous mutation in patients and control women at these restriction sites. CONCLUSION: The genetic analysis at our restriction sites in the SHP gene did not show any genetic variation in Korean PCOS patients. Our PCR-RFLP analysis was not covered the entire SHP gene (68 bp/ 1,006 bp), we need to further analysis of the entire SHP gene.

Intravenous immunoglobulin for patients with repeated failure of in vitro fertilization-embryo transfer / 대한산부인과학회잡지

OBJECTIVE: The implantation failure after embryo-transfer (ET) is a major continuing problem in in vitro fertilization (IVF). This study was undertaken to determine the effectiveness of intravenous immunoglobulin for treatment of individuals experiencing repeated unexplained in vitro fertilization-embryo transfer (IVF-ET) failure. METHODS: A total of nine consecutive infertile patients who failed to become pregnant after previous IVF-ET replacing at least three or more normal developed embryos each were included in our study. During the subsequent new IVF-ET cycle, each women received intravenous immunoglobulin 500mg/kg before the embryo transfer. RESULTS: Only one implantation occurred. There were no remarkable side effects. A specific effect of intravenous immunoglobulin for patients with repeated IVF-ET failure could not be demonstrated. CONCLUSION: High-dose intravenous immunoglobulin may not be useful for patients with repeated failure of embryo transfer.

A Case of Premature Ovarian Failure Assosiated with Gonadal Dysgenesis / 대한산부인과학회잡지

Prenature ovarian failure is a condition causing amenarrhea, hypoestrogenism, and elevated genadotropins in women younger than 40 years. A karyotype should be performed as part of basic laboratory evaluation for all patients with premature ovarian failure and prodromal premature ovarian failure. Development of a malignancy in a dysgenetic gonad is of major concern. The presence of a fragment of the Y chromosome is thought to be a key to the oncogenic potential of these gonads. The search for the testicular determining factor(TDF) has engendered much confusion about which part of the Y chromosome plays a role in malignancy. This was initially postulated to be the H- Y antigen. More recent data, however, localize the area near the centromere of the Y Chromosome, on the long arm(Yq). Malignant potential is clearly not linked to the testicular determining factor itself(SRY). This is a critical point in clinical medicine. Feilure to display SRY or a closely related sequence does not rule out the presence of the segment of the Y chromosome postulated to be associated with the development of malignancies. We have experienced a case of premature ovarian failure with chtomosomal abnormality involving Y chromosome fragment. So we report this case with a brief review of literatures.

Analysis of Chromosomal Abnormalities of Sperm from Patients with Advanced Age by FISH Method / 대한산부인과학회잡지

Determmation of the chomosomal constitution of human spermatozoa has been camed out though the human-hamster interspecific in vitro fertilization(IVF) system. In recent years, the introduction of fluorescence in-situ hybridization(FISH) technique has provided an alternative approach to evaluate the cbmmosomal constitution of human spermatozoa. The nuclei of mature spermatozoa are highly condensed with interpmtamine disulphide bridges, therefore the success of FISH on interphase human spermatozoa relies on partial decondensation of the sperm chromatin. In early studies, dithioothreitol(DTT) has been known as an efficient decondensation agent. Since then, several different decondensation methods using D1T have been establisdhed, and in terms of decondensation, we were tried to fix the optimal decondensation protocol using DlT. In our study, the optimal concentration and treatment time were 1-mM and 30 min, respectively. We examined chromosome complements of human sperm to investigate the effect of paternal age on the hequency of nondisjunction in human sperm. We investgated sperm karyotypes ftom two diffaent age groups)28+/-0.5, 46+/-6), A minimum of 1000 spermatozoa for one patient were analyzed. The mean frequencies of YY, XX, XY, 21-disamy spermatozoa ware 0.04%, 0.45%, 0.40%, 0.45% respectively in young age group and 1.06%, 0.62%, 1.06%, 0.76% in old ages. The mean frequency of disomy spermatozoa was higher in old age poup compare with those of young age group.

A Case of HAIR - AN Syndrome / 대한산부인과학회잡지

Polycystic ovary disease is a heterogenous endocrinopathy with many interacting causal factors. One potential such factor is chronic hyperinsulinemia. multiple, independent lines of evidence suppart the contention that chronic hyperinsulinemia causes ovarian hyperandragenism. This evidence includes: (1) mutations in the insulin receptor gene that cause severe hyperinsulinemia appear to be associated with ovarian hyperandrogenism, (2) insulin stimulates ovarian thecal and sttomal androgen seaetion in vitro, and (3) in some experimental models, manipulation of circulating insulin concentrations results in changes in circulating androgens. Although the association between hyperinsulinemia and hyperandrogenism remains to be fully explained at the molecular level, chronic hyperinsulinemia appears to be an important cause of hyperandrogenism. We have experienced a case of HAIR AN syndrome showing hyperandrogenism, insulin resistance and acanthosis nigricans in infertile patient. So we report this case with a brief review of literatures.

A Case of Azoospermia Associated with Y - Autosome Translocation / 대한산부인과학회잡지

Aneuploidy results from nondisjunction in either the meiotic division of the parents or the early cleavage divisions of the affected individuals. The sex chromosomes show a wide range of viable aneuploidy than do the autosomes. The incidence of 47,XXY and 47,XYY children increases with maternal age, as does that of autosomal trisomies, whereas the incidence of 45,X children does not increase with maternal age. In the group of sex chromosome aueuploidies, the 47,XXY and 47,XYY conditions occur with nearly equal hequency at birth. Translocations between X or Y chromosomes and an autosome or between an X chromosome and the Y chromosome cause sterility in human males. It has been assumed that a translocation involving either(or both) of the sex chromosomes would interfere with inactivation of the XY bivalent and thaeby disturb spermatogenesis. We bave experienced a case of Y-autosome translocation in azoospermic patient. So we report this case with a brief review of literatures.

Successful pregnancies using frozen thawed testicular sperm in ICSI program / 대한산부인과학회잡지

There are not much reports concerning with clinical results using frozen-thawed testivular sperm in ICSI program. It is speculated that the necessity of cryopreservation of testicular sperm to avoid repeating surgical procedure for obtaining sperm for ICSI. This study was carried out to confirm whether frozen-thawed testicular sperm could be fertilized and pregnancy could be achieved using embryos fertilized with frozen-thawed testicular sperm in ICSI program or not. Testicular sperm obtained from obstructive- or non-obstructive azoospermia patients were co-cultured for 3 days with Vero cells to improve sperm motility. By co-culturing with Vero cells for 3 days, O-ll% of sperm motility after thawing increased up to 8-42% after co-culturing. ICSI was performed using frozen-thawed, and co-cultured sperm with 66 oocytes obtained from 8 patients and 62 oocytes were survived and 49(79.0%) oocytes were fertilized normally. Embryo transfer was possible in 7 out of 8 patients, and pregnancy was achieved in 6 patients(85.7%). These results indicated that not only fresh testicular sperm but frozen-thawed testicular sperm can be used in ICSI program.

Preimplantation Genetic Diagnosis / 대한산부인과학회잡지

No abstract available.

Diagnosis of Fragile X Syndrome and DMD by using DIG System / 대한산부인과학회잡지

The aim of this study was to develop a rapid and safe non-radioactive DIG DNA labeling and detection for Southern blot analysis for fragile X syndrome and Duchenne muscular dystrophy (DMD). Southern blot analysis is accurate test showing expression of the (CGG)n repeat and abnormal methylation pattern of CpG island in hagile X syndrome, and good confirmative secondary test in case of deletion in DMD. But in terms of test rapidity, these conventional radioactive Southern analysis may not be feasible for rapid screening of prenatal samples and at-risk populations to determine their status and to provide genetic counseling to their families. As an alternative radioactive Southern blotting, DIG DNA labeling and detection system does not require handling of radioactive material nor require learning any new technology. The complete procedure of labeling the DNA and hybridization to detection of the first visible signal can be compbsbed witbin 7 days. In addition, hybridization solutions containing labeled DNA can be reused several times after renewed denaturation.

A Case of Associated with Autosomal Reciprocal Translocation / 대한산부인과학회잡지

Infertility, defined as 1 year of unprotected coitus without conception, affects approximately 10 to 15% for couples of reproductive age. Approximately 35% of these cases are attributable to male factor infertility. A major cause of male infertility is chromosome abnormality, such as 47 chromosomes with an XXY karyotype. Early surveys of infertile males showed that the incidence of major chromosome abnormality in infertile males in azoospermic patients. When patients are treated for male infertility, a chromosome analysis including a search for abnormality at the DNA level, should be performed. We have experienced a case of autosomal reciprocal translocation in azoospermic patient. So we report this case with a brief review of literatures.

Prenatal Cytogenetic Analysis of 145 Fetuses Conceived by Intracytoplasmic Sperm Injectiion ( ICSI ) / 대한산부인과학회잡지

Prenatal diagnoses were performed in 145 fetuses resulting from 73 singleton and 36 twin pregnancies, all established by intracytoplasmic sperm injection (ICS: amniocentesis in 108 patients and Chorionic villus sampling in one. The prenatal cytogenetic results were obtained from pregnancies after ICSI using ejaculated spermatozoa, epididymal spermatozoa, testicular spermatozoa and after the replacement of frozen-thawed embryos derived from ICSI. The Karyotypes were normal in 138 cases (95.2%) of the prenatal diagnoses and there were 2 cases (1.4%) de novo and 5 cases (3.4%) inherited chromosomal aberrations. The two cases of de novo abnormalities were: 46, XY, t(6;7)(q21;p22) and 47, XY, +21 (trisomy 21).

Midtrimester Amniocentesis-Cytogenetic Analysis of 1,274 Cases / 대한산부인과학회잡지

Diagnostic amniocentesis is one of the most useful technique for the prenatal detection of genetic disorders. Traditionally standard amniocentesis has been most commonly performed during the 2nd trimester from 16 to 20 weeks` gestation. Our laboratory has received 1,284 midtrimester amniotic fluid specimens during the past 5 year period for cytogenetic analysis and 1,274 were successfully cultured and yielded results. This study was based on data from 1,274 genetic amniocentesis performed at CHA General Hospital from Jan. 1991 to Dec. 1995. Chromosomal abnormalities were found in 61(4.8%) of the cases. There were 23 cases of aneuploidy, 37 cases of chromosomal rearrangemen t and 1 case of mosaicism.

Diagnosis of DMD/BMD by Multiplex PCR and Southern Blot Analysis / 대한산부인과학회잡지

Duchene and Becker muscular dystrophy(DMD/BMD) results from mutations in thedystrophin gene, and enormous genetic locus that spans more than two million base paris ofDNA on the human X chromosome. Some 60% of DMD patients exhibit deletions, which canbe found by cDNA hybridization or, were recently, by polymerase chain reaction analysis.We have used the multiplex PCR to identify deletion mutations in the human dystrophingene. By simultaneously amplifying genomic regions flanking 17 sepastrate exons inmutational hot spots, we were able to detect 16 exons in one family. The DNA encoding eachof the 17 exons in the dystrophin gene is copied a million fold to make it visible in anagarose gel. To be certain that the missing band is not artifact of the amplificationprocedure, the DNA from the blood sample was analyzed by Southern hybridization.