Yih-Ron Lien

Effects of cryopreservation on meiotic spindles of oocytes and its dynamics after thawing: clinical implications in oocyte freezing--a review article.

Embryo freezing has been a successful practice, but oocyte cryopreservation formerly achieved poorer results. This was mainly due to low rates of survival, fertilization, and development. The major dissimilarities for oocytes to embryos are the character of the plasma membrane, the presence of cortical granules, at the metaphase of meiosis II with the spindle system. In addition, the oocytes must be fertilized by sperm at the appropriate time. To improve the survival rate, a refined slow freezing method with increased sucrose concentration would dehydrate oocytes more sufficiently. Vitrification is another approach to prevent ice crystal formation. Intracytoplasmic sperm injection is used to overcome possible zona hardening from the release of cortical granules. The microtubules of meiotic spindles are vulnerable to the thermal changes and would depolymerize. Cryopreserved oocytes exhibited serious disturbances of the microtubules immediately after thawing. Fertilization of oocytes with disorganized spindles could lead to chromosomal aneuploidy, digyny, and arrest of cleavage. After incubation, the microtubules would repolymerize in a time-dependent way. Normal fertilization and development of cryopreserved oocytes improved after appropriate incubation and timing of insemination, compatible with recovery of the spindles. With the improvement of survival, fertilization, and cleavage, oocyte cryopreservation would gain an imperative role.

Cryopreservation of mature human oocytes by vitrification with ethylene glycol in straws

OBJECTIVE To examine the effect of vitrification with ethylene glycol (EG) for mature human oocytes in straws. DESIGN Prospective, randomized, in vitro experiments. SETTING Reproductive unit of a university hospital. PATIENT(S) Immature oocytes from 110 patients undergoing intracytoplasmic sperm injection (ICSI). INTERVENTION(S) The immature oocytes were incubated to reach metaphase II (MII). The MII oocytes were treated with EG-based cryoprotectants and vitrified in straws. They were diluted in sucrose solutions, inseminated by ICSI, and cultured in vitro. MAIN OUTCOME MEASURE(S) Survival, fertilization, and embryo cleavage. RESULT(S) The survival rates were greater for oocytes pretreated with 1.5 M of EG (65% for 0 minute, 93% for 5 minutes, and 96% for 10 minutes). The oocytes vitrified in 60 and 90 seconds had a greater rate of fertilization than those vitrified in 120 seconds. There were no differences in survival and fertilization for vitrified oocytes diluted by three or four steps. The cleavage rates to the six- to eight-cell stage were comparable with controls. However, no blastocyst formation was observed in vitrified oocytes. CONCLUSION(S) Vitrification of human oocytes with EG in straws achieves a high rate of survival, fertilization, and early cleavage of embryos. Further studies should be conducted for the improvement of blastocyst formation.

Transmission of de novo mutations of the deleted in azoospermia genes from a severely oligozoospermic male to a son via intracytoplasmic sperm injection

OBJECTIVE To investigate the transmission of microdeletions in the deleted in azoospermia (DAZ) genes to a male offspring via intracytoplasmic sperm injection (ICSI). DESIGN Case report. SETTING Reproductive unit of a university teaching hospital. PATIENT(S) A 29-year-old, severely oligozoospermic male with microdeletions of the DAZ genes in Yq interval 6 and his son, who was conceived via ICSI. INTERVENTION(S) DNA screening for the microdeletions in Yq interval 6 with 24 sequence tagged sites with the use of polymerase chain reaction amplification for the patient, the patients father, and the patients son. Paternity identification was performed using nine hypervariable short tandem repeats. MAIN OUTCOME MEASURE(S) Deletion mapping of Yq interval 6 from sequence tagged sites and electropherogram of short tandem repeats for DNA fingerprinting. RESULT(S) The son had the same microdeletions of the DAZ genes as the patient, and the patients father had normal DAZ genes. The paternity of the patient, the patients father, and the patients son was verified. CONCLUSION(S) De novo DAZ microdeletions in an infertile male can be transmitted to a male offspring via ICSI. DNA screening tests for DAZ genes before ICSI may help in the genetic counseling of patients with idiopathic azoospermia or severe oligozoospermia.

The duration of pre-ovulatory serum progesterone elevation before hCG administration affects the outcome of IVF/ICSI cycles

STUDY QUESTION During controlled ovarian stimulation (COS), does the duration of premature serum progesterone (P) elevation before administration of hCG affect the outcomes of IVF/ICSI embryo transfer (-ET) cycles? SUMMARY ANSWER The duration of the premature serum P elevation is inversely related to the clinical pregnancy rate of IVF/ICSI-ET cycles. WHAT IS KNOWN AND WHAT THIS PAPER ADDS The majority of the previous studies only considered a single serum P measurement made on the day of hCG administration and the results of attempts to relate this to IVF/ICSI-ET outcomes were controversial. However, the effect of the duration of premature serum P elevation before the hCG administration on the outcomes of IVF/ICSI-ET cycles has not been studied well. Here we demonstrate that the duration of premature serum P elevation has a more significant inverse correlation than the absolute serum P concentration on the day of hCG administration with IVF/ICSI-ET outcomes. DESIGN It is a retrospective, single-centre cohort study. A total of 1784 IVF and/or ICSI-ET cycles were included from October 2005 to June 2011. PARTICIPANTS AND SETTING A total of 1784 patients underwent their IVF and/or ICSI-ET cycles in a university hospital IVF unit. The inclusion criteria include (i) age between 20 and 42 years and (ii) eligible indications for COS before IVF/ICSI. MAIN RESULTS AND THE ROLE OF CHANCE The duration of premature serum P elevation to >1 ng/ml is significantly inversely associated with the probability of clinical pregnancy (odds ratio = 0.773, 95% confidence interval: 0.660-0.891, P < 0.001), after adjustment for possible confounders with multivariate logistic regression analysis. However, the significance of inverse correlation between the absolute serum P concentration on the day of hCG administration with clinical pregnancy rate decreased after adjustment. BIAS, CONFOUNDING AND OTHER REASONS FOR CAUTION The cutoff value we chose to define premature serum P elevation (P > 1.0 ng/ml) might not be able to be applied to different immunoassay kits and study population. The retrospective nature of this study inevitably might be influenced by some selection bias. GENERALIZABILITY TO OTHER POPULATIONS Older patients (>42 years) are excluded from our study.

Schedule to inject in vitro matured oocytes may increase pregnancy after intracytoplasmic sperm injection

To ascertain the value of using immature oocytes in an intracytoplasmic sperm injection (ICSI) program, the authors designed a schedule, at 5 p.m. on day 1 (the day of oocyte retrieval) and at 8 a.m. and 2 p.m. on day 2, to recognize and inject the in vitro matured (IVM) oocytes. For the 1,166 oocytes retrieved in 107 ICSI cycles, 128 (11.0%) were at the stage of metaphase I (MI) and 113 (9.7%) at germinal vesicle. Routine ICSI for metaphase 11 oocytes was performed at 2 p.m. on day 1 (initial ICSI). In culture medium of human tubal fluid with 15% maternal serum, 85.1% (205/241) immature oocytes progressed to maturation in which 16.4% (21/128) of MI oocytes matured at 5 p.m. of day 1. The rate of normal fertilization for IVM oocytes (58.5%) was not significantly different from that of initial ICSI (64.0%). One patient received a transfer of two fertilized IVM oocytes alone that were injected at 5 p.m. of day 1, maturing from the MI stage, and achieved a normal pregnancy. The fertilized IVM oocytes were replaced along with the embryos from initial ICSI for 40 cycles that led to 14 (35%) clinical pregnancies. In 43 fertilized IVM oocytes donated for research, we observed that cleavage (95.3%) to the 2- to 4-cell stage was not distinct from that of initial ICSI (94.6%); however, the percentage of embryos of grade I and II morphology was significantly smaller (24.4% vs. 62.5%). Only five (11.6%) developed to blastocysts in vitro. Twenty-one fertilized IVM oocytes were frozen for future transfer. A schedule to inject IVM oocytes in ICSI cycles may generate more accessible embryos for fresh transfer or cryopreservation to increase the chance of pregnancy, although the embryo quality was relatively poor.To ascertain the value of using immature oocytes in an intracytoplasmic sperm injection (ICSI) program, the authors designed a schedule, at 5 p.m. on day 1 (the day of oocyte retrieval) and at 8 a.m. and 2 p.m. on day 2, to recognize and inject the in vitro matured (IVM) oocytes. For the 1166 oocytes retrieved in 107 ICSI cycles, 128 (11.0%) were at the stage of metaphase I (MI) and 113 (9.7%) at germinal vesicle. Routine ICSI for metaphase II oocytes was performed at 2 p.m. on day 1 (initial ICSI). In culture medium of human tubal fluid with 15% maternal serum, 85.1% (205/241) immature oocytes progressed to maturation in which 16.4% (21/128) of MI oocytes matured at 5 p.m. of day 1. The rate of normal fertilization for IVM oocytes (58.5%) was not significantly different from that of initial ICSI (64.0%). One patient received a transfer of two fertilized IVM oocytes alone that were injected at 5 p.m. of day 1, maturing from the MI stage, and achieved a normal pregnancy. The fertilized IVM oocytes were replaced along with the embryos from initial ICSI for 40 cycles that led to 14 (35%) clinical pregnancies. In 43 fertilized IVM oocytes donated for research, we observed that cleavage (95.3%) to the 2- to 4-cell stage was not distinct from that of initial ICSI (94.6%); however, the percentage of embryos of grade I and II morphology was significantly smaller (24.4% vs. 62.5%). Only five (11.6%) developed to blastocysts in vitro. Twenty-one fertilized IVM oocytes were frozen for future transfer. A schedule to inject IVM oocytes in ICSI cycles may generate more accessible embryos for fresh transfer or cryopreservation to increase the chance of pregnancy, although the embryo quality was relatively poor.

Effect of conjugated equine estrogen in combination with two different progestogens on the risk factors of coronary heart disease in postmenopausal Chinese women in Taiwan: a randomized one‐year study

Background.  To compare the effect of hormone replacement therapy (HRT) using estrogen plus dydrogesterone or estrogen plus medroxyprogesterone acetate (MPA) on the risk factors for coronary heart disease (CHD) in postmenopausal women.

Update on management of ovarian hyperstimulation syndrome

Ovarian hyperstimulation syndrome (OHSS) is a relatively common complication of ovarian stimulation and can be life threatening. The pathophysiology of OHSS is characterized by increased capillary permeability, leading to leakage of fluid from the vascular compartment, with third-space fluid accumulation and intravascular dehydration. The increased intra-abdominal pressure indicated that OHSS may be considered a compartment syndrome. Vascular endothelial growth factor, also known as vascular permeability factor, has emerged as one of the mediators intrinsic to the development of OHSS. Conventional management is focused on supportive care until the spontaneous resolution of the condition. The standard of care for treatment-monitoring of appropriate clinical parameters, fluid balance management, thrombosis prophylaxis, and ascites treatment-should prevent severe morbidity in most cases. This review will cover inpatient and outpatient management. The potential therapeutic approach targeting the vascular endothelial growth factor system will be discussed.

Prospective comparison of short and long GnRH agonist protocols using recombinant gonadotrophins for IVF/ICSI treatments

This is a prospective comparative study investigating cost and effectiveness of IVF/ intracytoplasmic sperm injection (ICSI) treatments after stimulation with recombinant gonadotrophins following either the short or long gonadotrophin-releasing hormone (GnRH) agonist protocol. Patients in the short protocol (n = 120) were administered buserelin nasal sprays from day 2 of the menstrual cycle and recombinant FSH from day 5. Patients in the long protocol (n = 120) were administered buserelin from the previous mid-luteal phase and recombinant FSH after achieving down-regulation. The average age and basal FSH concentrations of both groups were similar. The serum LH concentrations during ovarian stimulation were significantly higher with the short protocol. The total cost of recombinant gonadotrophins (US

Tuboovarian Abscesses in Postmenopausal Women

527 +/- 184 versus US

Twin Pregnancies Conceived by Assisted Reproductive Technology: Maternal and Perinatal Outcomes

795 +/- 244, P < 0.001) was significantly lower in the short protocol, but there was no significant difference in delivery rates (47.5 versus 36.7%) between the short and long protocols. LH flare-up during the short protocol does not seem to impair the treatment outcome. Using recombinant gonadotrophins, the short GnRH agonist protocol is an effective and cheaper choice for IVF/ICSI treatments.