Lipidomics profiles of human spermatozoa: insights into capacitation and acrosome reaction using UPLC-MS-based approach.

Introduction: Lipidomics elucidates the roles of lipids in both physiological and pathological processes, intersecting with many diseases and cellular functions. The maintenance of lipid homeostasis, essential for cell health, significantly influences the survival, maturation, and functionality of sperm during fertilization. While capacitation and the acrosome reaction, key processes before fertilization, involve substantial lipidomic alterations, a comprehensive understanding of the changes in human spermatozoa's lipidomic profiles during these processes remains unknown. This study aims to explicate global lipidomic changes during capacitation and the acrosome reaction in human sperm, employing an untargeted lipidomic strategy using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). Methods: Twelve semen specimens, exceeding the WHO reference values for semen parameters, were collected. After discontinuous density gradient separation, sperm concentration was adjusted to 2 x 106 cells/ml and divided into three groups: uncapacitated, capacitated, and acrosome-reacted. UPLC-MS analysis was performed after lipid extraction from these groups. Spectral peak alignment and statistical analysis, using unsupervised principal component analysis (PCA), bidirectional orthogonal partial least squares discriminant analysis (O2PLS-DA) analysis, and supervised partial least-squares-latent structure discriminate analysis (PLS-DA), were employed to identify the most discriminative lipids. Results: The 1176 lipid peaks overlapped across the twelve individuals in the uncapacitated, capacitated, and acrosome-reacted groups: 1180 peaks between the uncapacitated and capacitated groups, 1184 peaks between the uncapacitated and acrosome-reacted groups, and 1178 peaks between the capacitated and acrosome-reacted groups. The count of overlapping peaks varied among individuals, ranging from 739 to 963 across sperm samples. Moreover, 137 lipids had VIP values > 1.0 and twenty-two lipids had VIP > 1.5, based on the O2PLS-DA model. Furthermore, the identified twelve lipids encompassed increases in PI 44:10, LPS 20:4, LPA 20:5, and LPE 20:4, and decreases in 16-phenyl-tetranor-PGE2, PC 40:6, PS 35:4, PA 29:1, 20-carboxy-LTB4, and 2-oxo-4-methylthio-butanoic acid. Discussion: This study has been the first time to investigate the lipidomics profiles associated with acrosome reaction and capacitation in human sperm, utilizing UPLC-MS in conjunction with multivariate data analysis. These findings corroborate earlier discoveries on lipids during the acrosome reaction and unveil new metabolites. Furthermore, this research highlights the effective utility of UPLC-MS-based lipidomics for exploring diverse physiological states in sperm. This study offers novel insights into lipidomic changes associated with capacitation and the acrosome reaction in human sperm, which are closely related to male reproduction.

Impact of SARS-CoV-2 infection on clinical outcomes of in vitro fertilization treatments: a systematic review and meta-analysis.

The influence of SARS-CoV-2 infection on clinical outcomes in patients undergoing in vitro fertilization has been uncertain. Therefore, this systematic review and meta-analysis aimed to evaluate the impact of past SARS-CoV-2 infection on IVF outcomes. A comprehensive search of PubMed, EMBASE, and Cochrane Library databases was conducted from December 2019 to January 2023. Included studies comparing IVF outcomes between patients with prior SARS-CoV-2 infection and controls without previous infection were analyzed. Study quality was assessed using the Newcastle-Ottawa Quality Assessment Scale. Sensitivity analysis, publication bias, and heterogeneity were also examined. The review protocol was registered with PROSPERO (CRD42023392007). A total of eight studies, involving 317 patients with past SARS-CoV-2 infection and 904 controls, met the inclusion criteria. The meta-analysis revealed no significant differences between the infection group and controls in terms of clinical pregnancy rate (OR 0.97, 95% CI 0.73-1.29; P = 0.82), implantation rate (OR 0.99, 95% CI 0.67-1.46; P = 0.96), or miscarriage rate (OR 0.64, 95% CI 0.15-2.65; P = 0.53). Subgroup analyses based on transfer type demonstrated comparable clinical pregnancy rates between the two groups in both fresh embryo transfer (OR 0.97, 95% CI 0.69-1.36; P = 0.86) and frozen embryo transfer (OR 0.96, 95% CI 0.38-2.44; P = 0.94). In conclusion, this meta-analysis suggests that previous SARS-CoV-2 infection does not have a detrimental impact on clinical outcomes in IVF patients. These findings provide valuable insights into assessing the influence of prior SARS-CoV-2 infection on successful pregnancy outcomes in IVF treatment. The systematic review was performed based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. This review was prospectively registered with the International Prospective Register of Systematic Reviews (ID CRD42023392007) on January 16, 2023.

Applications of laser technology in the manipulation of human spermatozoa.

The application of laser technology in the field of assisted reproductive technology (ART) has experienced rapid growth over the past decades owing to revolutionary techniques such as intracytoplasmic sperm injection (ICSI), preimplantation genetic testing (PGT), and in vitro manipulation of gametes and embryos. For male gametes, in vitro manipulation techniques include spermatozoa selection, sorting, immobilization, and quality assessment. A number of studies have been conducted to investigate the application of different laser technologies in the manipulation of human spermatozoa. However, there is a lack of a unified understanding of laser application in the in vitro manipulation of sperm and safety considerations in ART and, subsequently, the inability to make clear and accurate decisions on the clinical value of these laser technologies. This review summarizes the advancements and improvements of laser technologies in the manipulation of human spermatozoa, such as photobiomodulation therapy, laser trap systems for sperm analysis and sorting, laser-assisted selection of immotile sperm and laser-assisted immobilization of sperm prior to ICSI. The safety of those technologies used in ART is also discussed. This review will provide helpful and comprehensive insight into the applications of laser technology in the manipulation of human spermatozoa.

Clinical outcomes of frozen-thawed blastocysts from zygotes with no or one pronucleus for in vitro fertilization and intracytoplasmic sperm injection cycles.

PURPOSE: To investigate the clinical outcomes of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles using frozen-thawed blastocyst transfers derived from zygotes with no (0PN) or one pronucleus (1PN). METHODS: This retrospective study included 7084 0PN, 2238 1PN, and 72,266 two pronuclear (2PN) embryos cultured to the blastocyst stage from 19,631 IVF and 12,377 ICSI cycles between March 2018 and December 2021. Developmental potential and clinical outcomes of 0PN, 1PN, and 2PN embryos were analyzed. A total of 290 0PN-, 92 1PN-, and 1906 2PN-derived single frozen-thawed blastocyst transfers were performed. Chromosome euploid rates of 0PN-, 1PN-, and 2PN-derived blastocysts were analyzed by next-generation sequencing. Euploid 0PN- and 1PN-derived blastocysts underwent subsequent Infinium Asian Screening Array gene chip analysis to detect ploidy alterations. RESULTS: Available blastocyst rates of 0PN and 1PN embryos were significantly lower than those of 2PN embryos in both IVF and ICSI cycles. Single 0PN and 1PN blastocysts transferred in frozen-thawed cycles resulted in a similar clinical pregnancy rate, miscarriage rate, live birth rate, and neonatal outcome to 2PN blastocysts in IVF and ICSI cycles. Genetic analysis showed that euploid rates of 0PN- and 1PN-derived blastocysts used for ICSI cycles were similar to that of 2PN-derived blastocysts. CONCLUSION: Our study indicated that 0PN- and 1PN-derived blastocysts resulted in similar clinical outcomes to 2PN-derived blastocysts. The 0PN- and 1PN-derived blastocysts from ICSI cycles can be transferred as well as those from IVF cycles when the number of 2PN-derived blastocysts is insufficient.

Pregnancy outcomes following in vitro fertilization treatment in women with previous recurrent ectopic pregnancy.

OBJECTIVES: The aim of this study was to investigate the impact of a history of recurrent ectopic pregnancy (EP) on pregnancy outcomes of subsequent in vitro fertilization (IVF) treatment. METHODS: A retrospective cohort study involving 457 women with a history of recurrent EP (REP group), 912 women with a history of single EP (SEP group), and 1169 women with a history of intrauterine pregnancy (IUP group) as the control group, was conducted. IVF outcomes were compared for each cohort. RESULTS: The incidence of EP in the REP group after IVF treatment was significantly lower than those in the SEP group (2.4% vs. 6.8%, P = 0.011), and similar to those in the IUP group (2.4% vs. 2.1%, P = 0.830). No significant differences were observed in the clinical pregnancy rate, miscarriage rate, and live birth rate among the three groups. There was no statistically significant difference in the recurrent EP rate between the salpingectomy and salpingostomy treatments. Adjusting for maternal and treatment factors did not influence live birth rates for women with previous REP compared with women with previous SEP and those with IUP. The odds of EP were 82.2% lower (OR 0.178, 95% CI 0.042-0.762; P = 0.020) in women who had blastocyst transfer compared with cleavage embryo transfer in the SEP group. The odds of EP were over six times (OR 6.260, 95% CI 1.255-31.220; P = 0.025) in women who underwent double embryo transfer as opposed to single embryo transfer in the IUP group. CONCLUSION: Our results indicate that women with previous recurrent EP have a lower risk of EP after IVF in comparison with women with previous single EP. Previous EP has no significant adverse effect on the main IVF outcomes. The salpingostomy and salpingectomy treatments of EP do not significantly affect the incidence of recurrent EP after IVF.

Time to pregnancy in women with previous ectopic pregnancy undergoing in vitro fertilization treatment: a retrospective cohort study.

We aimed to investigate the difference in the time to pregnancy (TTP) between women with previous ectopic pregnancy (EP) and control women following in vitro fertilization (IVF) treatment and the association between TTP and the number of oocytes retrieved and embryos available. A retrospective study involving 1097 women, 547 of which had previous EP and 550 were control women whose previous pregnancy were abortion, was conducted. Women in the EP group had significantly longer median TTP than those in the control group (36; range, 12-252 vs 28; range, 12-220; P = 0.019). For women with previous EP, > 48 months TTP was most likely associated with low numbers of oocytes retrieved and embryos available compared to TTP of ≤ 24 months or 25-48 months, and women with younger age had a shorter TTP, higher numbers of oocytes retrieved and embryos available. A Cox proportional hazards model showed that maternal age was significantly related to the pregnancy over the TTP (adjusted hazard ratio, 0.934; P < 0.001). In conclusion, women with previous EP have a significantly increased TTP than control women with previous abortion. For women with previous EP, TTP is negatively associated with the numbers of oocytes retrieved and embryos available.

Gene mutations associated with fertilization failure after in vitro fertilization/intracytoplasmic sperm injection.

Fertilization failure during assisted reproductive technologies (ART) is often unpredictable, as this failure is encountered only after in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) have been performed. The etiology of fertilization failure remains elusive. More and more mutations of genes are found to be involved in human fertilization failure in infertile patients as high throughput sequencing techniques are becoming widely applied. In this review, the mutations of nine important genes expressed in sperm or oocytes, PLCZ1, ACTL7A, ACTL9, DNAH17, WEE2, TUBB8, NLRP5, ZP2, and TLE6, were summarized and discussed. These abnormalities mainly have shown Mendelian patterns of inheritance, including dominant and recessive inheritance, although de novo mutations were present in some cases. The review revealed the crucial roles of each reported gene in the fertilization process and summarized all known mutations and their corresponding phenotypes. The review suggested the mutations might become promising targets for precision treatments in reproductive medicine. Moreover, our work will provide some helpful clues for genetic counseling, risk prediction, and optimizing clinical treatments for human infertility by supplying the useful and timely information on the genetic causes leading to fertilization failure.

The association of rs11457523 in HSP90AA1 with idiopathic male infertility in the Chinese population.

The association of single nucleotide polymorphisms (SNPs) in heat shock protein 90 (HSP90) genes with idiopathic male infertility remains unclear. In this study, the five selected SNPs in HSP90AA1 namely rs10133307, rs10873531, rs11547523, rs11621560 and rs7145597 were genotyped in 116 idiopathic infertile males and 185 ethnically matched fertile males using the Sequenom MassARRAY assay. The role of these SNPs in male infertility was then studied using multiple genetic models. We observed that genotype distribution (p = .028) and allelic frequency (p = .032) of rs11547523 were significantly different between the infertile and fertile groups. In particular, A genotype of rs11547523 was associated with an increased risk of infertility in the allele (OR = 2.508, p = .048), dominant (OR = 2.733, p = .030) and additive models (OR = 0.366, p = .031). However, there were no significant differences in semen parameters including seminal volume (p = .452), sperm concentration (p = .727), total sperm number (p = .588), motility (p = .282) and morphology (p = .975) between A and A/G genotypes of rs11547523. These results indicate that rs11547523 in HSP90AA1 may be associated with idiopathic male infertility in the Chinese population. The outcome of this study contributes to the development of the diagnosis of male infertility.

Association of the duration of post-thaw culture with clinical outcome after vitrified-warmed day 3 embryo transfer in 10,464 cycles: A retrospective cohort study.

This study aimed to investigate the effect of the duration of embryo culture on clinical outcome in vitrified-warmed cycles.This retrospective cohort study enrolled 10,464 infertile patients, with a total of 18,843 vitrified-warmed day 3 embryos from 2012 to 2017 at a single center. The patients were divided into 2 groups: 9470 cycles in the short-term culture group (0.5-8 hours of post-thaw culture) and 994 cycles in the 48 to 72 hours culture group. The independent effect of the following variables on clinical outcomes was determined: duration of post-thaw culture, maternal age, transferred embryos, embryo quality, and endometrial thickness.We found that the pregnancy rate was positively associated with the post-thaw culture time. Ordinary least square regression analyses showed that the duration of post-thaw culture was positively associated with implantation and live birth rates overall. However, the implantation and live birth rates were not significantly associated with the post-thaw culture time in the short-term culture group. Additionally, maternal age and the number of transferred embryos were independent predictors of the implantation and live birth rates. Moreover, the duration of post-thaw culture did not affect live birth weight.These results indicated that the pregnancy rate is positively associated with the duration of post-thaw culture. Therefore, under the condition of not affecting work shifts, properly prolonging the duration of post-thaw culture to improve the outcome of frozen-thawed embryo transfer should be considered.

The effect on twinning rate of transferring double vitrified-warmed embryos in women of advanced reproductive age: a retrospective study.

Twin pregnancies are associated with greater risk of neonatal morbidity and mortality than a singleton. This study was performed to investigate the twin pregnancy rate when two vitrified-warmed embryos are transferred in women of advanced reproductive age (≥35 years at the time of oocyte retrieval) and to evaluate the implications of findings in selecting candidates for elective single embryo transfer (eSET). A retrospective analysis of data which included 2,038 women aged 35-45 years, who underwent vitrified-warmed double embryo transfer (DET), from January 2013 to December 2016 was undertaken. Pregnancy and twin rates were estimated after stratifying by prognostic profile. The twin pregnancy rate was lower in women with poor prognosis (12/96, 12.5%) as compared with that in women with favorable prognosis (102/374, 27.3%) and average prognosis (78/346, 22.5%) with significant differences (P < 0.05). The twin rate for women with favorable prognosis was 29.2% (70/240) in the cycles of women aged 35-37 years, 26.8% (26/97) in the cycles of women aged 38-40 years and 16.2% (6/37) in the cycles of women >40 years. The twin rate for women with average prognosis was 25.8% (51/198) in the cycles of women aged 35-37 years, 22.0% (22/100) in the cycles of women aged 38-40 years and 10.4% (5/48) in the cycles of women >40 years. The twin rate for women with poor prognosis was 15.3% (9/59) in the cycles of women aged 35-37 years, 10.3% (3/29) in the cycles of women aged 38-40 years and 0% (0/8) in the cycles of women >40 years. From these results, it was concluded that women with a favorable or average prognosis have a high risk of twin pregnancies. The finding can be used to guide future practice: that is, performing eSET in women with favorable or average prognosis and DET in women with poor prognosis.

Freeze-all embryo strategy in poor ovarian responders undergoing ovarian stimulation for in vitro fertilization.

This retrospective study determined the efficacy of the 'freeze-all' embryo strategy in poor ovarian responders undergoing ovarian stimulation for in vitro fertilization (IVF). A total of 559 poor responders who met Bologna criteria between January 2012 and December 2014 were included in this study: 256 in the fresh embryo transfer group and 303 in the freeze-all group. Vitrification and warming of day 3 embryos were performed using the Cryotop method. The poor responders treated with fresh embryo transfer and those treated with freeze-all strategy showed similar live birth rates per cycle (12.1% vs. 16.2%, p = .172) and per transfer (15.9% vs. 20.9%, p = .182). Multivariate logistic regression analysis showed that maternal age at retrieval (odds ratio, 0.919; 95% confidence interval, 0.865-0.977; p = .006) and number of good-quality embryos transferred (odds ratio, 1.953; 95% confidence interval, 1.346-2.835; p < .001) were significantly associated with the live birth rate. Freeze-all cycle is an acceptable treatment in poor ovarian responders, and it should be suggested by physicians as an alternative to cycle cancelation in case in which a fresh transfer would not be advantageous.

Slow freezing should not be totally substituted by vitrification when applied to day 3 embryo cryopreservation: an analysis of 5613 frozen cycles.

PURPOSE: This study aimed to compare slow freezing (SF) and vitrification (VT) techniques for day 3 embryo cryopreservation in infertile couples. METHODS: This retrospective cohort study enrolled 5613 infertile patients, with 7862 frozen-thawed day 3 embryos and 3845 vitrified-warmed day 3 embryos, from 2010 to 2014, at a single center. The rates of embryo survival, pregnancy, implantation, miscarriage, live birth, and live birth weight were compared between the two groups. RESULTS: A total of 5613 cycles with 5520 transfers were analyzed. Using SF, the rates of overall embryo survival and fully intact blastomeres were lower than those in VT (91.5 vs. 97.4 % and 68.7 vs. 92.3 %, respectively). The rate of good quality embryos after thawing/warming was lower in SF than in VT. In single frozen embryo transfer cycles (FETs), the pregnancy and implantation rates were similar between the two groups (35.0 vs. 40.8 % and 34.6 vs. 35.9 %, respectively). In double FETs, the pregnancy rate per cycle was also similar between the groups (58.8 vs. 58.4 %). The implantation rate per embryo transfer was significantly higher with SF than with VT (38.8 vs. 34.6 %). With adjustment for maternal age and the number of good quality embryos, differences in implantation rate remained significant (adjusted P value, SF vs. VT P < 0.05). No independent effect was found for the method of cryopreservation on the pregnancy rate. No significant differences in the rates of miscarriage, live birth, and live birth weight were observed between the two techniques. CONCLUSIONS: Despite the significantly low embryo survival rate, fully intact blastomere rate, and good quality embryo rate in SF, the pregnancy and implantation rates were not adversely affected in single and double FETs. SF yielded an equivalent miscarriage rate, live birth rate, and live birth weight compared with VT. The SF protocol to cryopreserve day 3 embryos still should be considered.

S100P regulates trophoblast-like cell proliferation via P38 MAPK pathway.

S100P was originally isolated from the placenta, and is expressed in very high levels in trophoblast cells, but its role on trophoblast cells proliferation has not yet been studied. In this study, we aimed to investigate the potential role of S100P in human placental development, and the impact of its expression regulation on cellular function as well as molecular mechanisms involved in trophoblast-like cells. We found that the expression of S100P in first trimester placenta was significantly reduced in spontaneous abortion patients with respect to normal pregnant women. Up-regulation of S100P in JAR cells promoted JAR cells proliferation, and increased the expression of phosphorylated P38 (p-P38) mitogen-activated protein kinase (MAPK) and p-ERK MAPK. However, the effects of S100P on JAR cells proliferation were prevented by P38 inhibitor-SB203580, but not by ERK inhibitor-PD98059. These results showed that S100P may have a physiological role in normal pregnant development, and regulate trophoblast-like cell proliferation via modulating the P38 MAPK pathway.

Intracytoplasmic Sperm Injection Fertilization Rate Does Not Depend on the Proportion of Round-headed Sperm, Small-acrosomal Sperm, or Morphologically Normal Sperm in Patients with Partial Globozoospermia.

BACKGROUND: Generally, intracytoplasmic sperm injection (ICSI) may be the preferable method to treat partial globozoospermia, but whether there exist some correlations between ICSI fertilization rate and the proportion of round-headed sperm or morphologically normal sperm remains open. This study was to explore the correlation between ICSI fertilization rate and the sperm morphology in patients with partial globozoospermia. METHODS: Thirty-four patients diagnosed with partial globozoospermia accepted the following assisted fertilization treatments - 2 cases accepted in-vitro fertilization (IVF) alone, 26 cases accepted ICSI alone, and 6 accepted split IVF/ICSI. Detailed morphological characteristics were described using Diff-Quik rapid staining. Sixty cases accepting IVF or ICSI treatment in our reproductive center were considered as the control group after being matched by relevant criteria. Fertilization rate, embryo quality, embryo implantation rate and clinical pregnancy rate were calculated. RESULTS: Besides very high proportion of round-headed sperm, partial globozoospermia also showed very high proportion of small-acrosomal sperm and very low proportion of morphologically normal sperm. Fertilization rate of IVF (IVF alone plus split IVF) was very low in partial globozoospermia (25.4% ± 17.4%), but ICSI (ICSI alone plus split ICSI) achieved satisfying fertilization rate compared with the control group (66.2% ± 22.5% vs. 68.8% ± 29.4%, P > 0.05). In patients with partial globozoospermia, there were no correlations between ICSI fertilization rate and the proportion of round-headed sperm, small-acrosomal sperm, or morphologically normal sperm. CONCLUSIONS: There was high proportion of small-acrosomal sperm in partial globozoospermia. For patients with partial globozoospermia, ICSI is more preferable than IVF. ICSI fertilization rate does not depend on the proportion of round-headed sperm, small-acrosomal sperm, or morphologically normal sperm.

Heat shock protein 90 has roles in intracellular calcium homeostasis, protein tyrosine phosphorylation regulation, and progesterone-responsive sperm function in human sperm.

Heat shock protein 90 plays critical roles in client protein maturation, signal transduction, protein folding and degradation, and morphological evolution; however, its function in human sperm is not fully understood. Therefore, our objective in this study was to elucidate the mechanism by which heat shock protein 90 exerts its effects on human sperm function. By performing indirect immunofluorescence staining, we found that heat shock protein 90 was localized primarily in the neck, midpiece, and tail regions of human sperm, and that its expression increased with increasing incubation time under capacitation conditions. Geldanamycin, a specific inhibitor of heat shock protein 90, was shown to inhibit this increase in heat shock protein 90 expression in western blotting analyses. Using a multifunctional microplate reader to examine Fluo-3 AM-loaded sperm, we observed for the first time that inhibition of heat shock protein 90 by using geldanamycin significantly decreased intracellular calcium concentrations during capacitation. Moreover, western blot analysis showed that geldanamycin enhanced tyrosine phosphorylation of several proteins, including heat shock protein 90, in a dose-dependent manner. The effects of geldanamycin on human sperm function in the absence or presence of progesterone was evaluated by performing chlortetracycline staining and by using a computer-assisted sperm analyzer. We found that geldanamycin alone did not affect sperm capacitation, hyperactivation, and motility, but did so in the presence of progesterone. Taken together, these data suggest that heat shock protein 90, which increases in expression in human sperm during capacitation, has roles in intracellular calcium homeostasis, protein tyrosine phosphorylation regulation, and progesterone-stimulated sperm function. In this study, we provide new insights into the roles of heat shock protein 90 in sperm function.

The effect of progesterone level prior to oocyte retrieval on the numbers of oocytes retrieved and embryo quality in IVF treatment cycles: an analysis of 2,978 cycles.

PURPOSE: The study was designed to evaluate the relationship between serum progesterone (P4) response after hCG administration and the number of oocytes retrieved and the embryo quality in fresh IVF cycles. METHODS: We conducted a retrospective cohort study of women aged 24-43 years who underwent first fresh IVF cycle from 2011 to 2013 at a single practice. We compared the post-hCG serum P4 level with values on the day of hCG trigger. Patients were analyzed in long and short protocols independently. In addition, patients were stratified by post-hCG P4 response. Number of oocytes retrieved and embryo quality were the primary outcomes of interest. Ordinary least square regression models and logistic regression analysis models were created to identify predictive factors associated with embryological outcomes while adjusting for potential confounders. RESULTS: Among the 2,978 IVF cycles, 2,484 patients were in long protocols, and 494 patients were in short protocols. After adjusting for patient age, rFSH duration, and basal FSH levels, the associations between P4 response after hCG administration and number of oocytes retrieved (P < 0.001) remained statistically significant in both long and short protocols. Additionally, mature oocyte rate, fertilization rate, good quality embryo rate, pregnancy rate and implantation rate were not significantly associated with the P4 increase when adjusting for the same factors. However, pregnancy rate and implantation rate from frozen-thawed cycles increased gradually across the seven groups. CONCLUSIONS: Post-hCG P4 levels were positively associated with the number of oocytes retrieved, but did not affect oocyte or embryo quality. Our study suggests that the change in the post-hCG P4 level is another parameter that can be used by clinicians to assess the number of oocytes retrieved, and may further to estimate the pregnancy rate and live birth rate indirectly.

Effect of vitrification versus slow freezing of human day 3 embryos on ß-hCG levels.

PURPOSE: The study was designed to investigate the effect of vitrification and slow freezing for the cryopreservation of human day 3 embryos on serum ß-hCG levels in pregnancies established after frozen embryo transfer (FET). METHODS: Of the 1384 FET cycles initiated, 1131 embryo transfers met study criteria and assigned to one of two groups: 797 slow-freezing embryo transfers or 334 vitrified embryo transfers. Median values of ß-hCG and outcome of all pregnancies were compared between the two groups. Predictive values of serum ß-hCG on day 12 after embryo transfer for establishing ongoing pregnancy and pregnancy failure were determined by receiver operating characteristic (ROC) curve analysis. RESULTS: In the slow-freezing group, 383 ongoing pregnancies occurred (48.1 %), and transfers of vitrified embryos resulted in 154 pregnancies (46.1 %). Median ß-hCG values (279.2 IU/L) for ongoing pregnancies after transfer of vitrified embryos were significantly lower than that of slow frozen embryos (320.5 IU/L). The median values of ß-hCG for singleton in the two groups was statistically significant (P <0.05). For slow-freezing embryo transfers, the cut-off value of ß-hCG in predicting ongoing pregnancy was 147 IU/L (sensitivity 88.3 %, specificity 80.7 %). For vitrified embryo transfers, the value was 135 IU/L (sensitivity 84.4 %, specificity 76.3 %). CONCLUSIONS: Day 12 ß-hCG levels after FET are significantly affected by the methods of embryo cryopreservation for ongoing pregnancies. Furthermore, when using ß-hCG cut-off value to assess pregnancy outcome, the cryopreservation methods should be taken into account.

Effect of cumulus cell removal 4 h post-insemination on fertilization and embryo quality: a prospective randomized sibling-oocyte study.

PURPOSE: The study was designed to evaluate whether cumulus cell removal 4 h post-insemination could influence fertilization and embryo quality. METHODS: The study included 61couples undergoing standard long down regulation protocol from July 2011 to May 2012. Sibling oocytes of each patient were randomly assigned to either the 4 h group or the 20 group. For the 4 h group, cumulus cells were removed 4 h after gamete coincubation; for the 20 group, cumulus cells removal was performed 20 h after insemination. Fertilization rate, embryo quality, pregnancy rate and implantation rate were assessed. RESULTS: A total of 801 sibling cumulus-oocyte complexes (COCs) were randomized to the 4 h group (421 COCs) or 20 h group (380 COCs). There was no difference in the two pronuclei, one pronucleus and grade 1-2 embryo rate. Three pronuclei rate was significantly higher in the 4 h group compared to the 20 h group (12.6 % vs. 8.2 %, P = 0.041). Comparison of embryo transfer cycles in which either embryos from the 4 h group or 20 h group were transferred did not reveal any statistically significant differences in pregnancy or implantation rates. CONCLUSION: The results of the present study indicate that cumulus cell removal 4 h post-insemination may increase the percentage of tripronuclear zygotes. However, normal fertilization rate, embryo development, clinical pregnancy rate and implantation rates are not influenced by the timing of cumulus cell removal.

Prognostic value of total testosterone for pregnancy during treatment in patients with clomiphene-citrate-resistant polycystic ovary syndrome: a pilot study.

PURPOSE: Reduction of serum total testosterone (TT) is associated with pregnancy rate in polycystic ovary syndrome (PCOS) women receiving metformin, but most of the studies focus on the changes of basal levels of TT. The aim of this study was to evaluate whether the TT level around the ovulation period is related to the outcome of pregnancy in women with PCOS. METHODS: In total, 30 non-obese PCOS women with clomiphene citrate (CC) resistance from the Medical College's Reproductive Health Center were enrolled and randomly assigned to be treated with placebo (Group 1) or metformin (850 mg) (Group 2) twice daily for 3 months as the pre-treatment. Then, metformin alone was administered with CC, human menopausal gonadotropin (HMG) and human chorionic gonadotropin (HCG) to induce ovulation for 3 months in Group 1. In Group 2, CC/HMG/HCG was used to induce ovulation for 3 months without metformin. Follicle-stimulating hormone, luteinizing hormone, estradiol and TT levels before and after ovulation in pregnant cycles and non-pregnant cycles were evaluated over the course of treatment. RESULTS: A total of 26 subjects completed 65 cycles. The TT levels after ovulation in the pregnant cycles were significantly lower than in the non-pregnant cycles in both groups (P = 0.001 and P < 0.001, respectively). CONCLUSIONS: The level of TT after ovulation may be of prognostic value for pregnancy in non-obese women with PCOS and CC resistance during treatment.

[Calmodulin antagonists are involved in mouse sperm capacitation].

OBJECTIVE: To investigate the possible involvement of calmodulin in mouse sperm capacitation. METHODS: Calmodulin antagonists W7 at the concentrations of 50, 100 and 200 micromol/L and calmidazolium (CZ) at the concentrations of 10, 20 and 30 micromol/ L, were coincubated with mouse sperm for 2 hours, respectively. The percentage of pattern B sperm was measured by chlorotetracycline staining. Then the sperm were coincubated with 100 micromol/L W7 or 10 micromol/L calmidazolium (CZ) before acrosome reaction was induced by 5 micromol/L progesterone and evaluated by the same method. RESULTS: After the treatment of the sperm with different concentrations of CZ or W7, the percentages of pattern B sperm decreased in a dose-dependent manner, significantly different from the control (P < 0.05). There was a statistic difference in the rate of acrosome reaction between the experiment and the control group (P < 0.01). CONCLUSION: Calmodulin is a key protein involved in mouse sperm capacitation.