The correlation between sperm percentage with a small acrosome and unexplained in vitro fertilization failure.

PURPOSE: Since the unexplained in vitro fertilization failure occurs frequently, it is of great importance and clinical value to identify potential underlying predictors. This study aimed to explore whether the percentage of sperm with a small acrosome was correlated with unexplained in vitro fertilization failure. METHODS: A new acrosomal function evaluation index (the percentage of sperm with a small acrosome) was introduced into the analysis of sperm morphology. The association between the index and acrosome function by acrosin activity detection test and acrosome reaction test was investigated. In addition, the correlation with unexplained in vitro fertilization failure was further explored. Finally, the ROC curve was used to analyze the diagnostic efficacy on the failure of in vitro fertilization and the cutoff value was calculated. RESULTS: As the increasing of the percentage of sperm with a small acrosome, the value of acrosin activity, acrosome reaction rate, and in vitro fertilization rate were reduced, with a statistically significant difference (P < 0.05). The index in the low fertilization rate group was significantly higher than that in the normal fertilization rate group (P < 0.05). Finally, the results of ROC curve found that when the index was 43.5%, the sensitivity and specificity were 74.2% and 95.3%, respectively. CONCLUSION: The percentage of sperm with a small acrosome was positively correlated with unexplained in vitro fertilization failure, which could be potentially used as a prognostic index for the failure of in vitro fertilization. TRIAL REGISTRATION: [Ethics review acceptance No IIT20210339B].

Value of serum and follicular fluid sirtuin (SIRT)1 and SIRT2 protein levels in predicting the outcome of assisted reproduction.

BACKGROUND: To explore whether serum and follicular fluid (FF), sirtuin 1 (SIRT1), and SIRT2 could predict the outcome of assisted reproduction. METHODS: All patients underwent in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) for the first time in the Reproductive Medicine Center of the First Affiliated Hospital of Zhejiang University Medical College from March 2018 to December 2018. According to cumulative clinical pregnancy outcomes, the patients were divided into a pregnancy group and non-pregnancy group. We measured the serum levels of SIRT1, SIRT2, anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) from the second to the fifth day of menstruation, and the levels of SIRT1 and SIRT2 in serum and FF on the day of human chorionic gonadotropin (HCG) injection and oocyte retrieval. RESULTS: A total of 125 patients met the inclusion criteria. The pregnancy group comprised 56 cases and non-pregnancy group 69 cases. There were significant differences in basal level SIRT2 (bSIRT2), AMH, antral follicle count (AFC), number of oocytes obtained, number of mature eggs, number of fertilized eggs, number of excellent embryos, number of blastocyst formations, and number of transferred high-quality embryos between the two groups. The area under the curve (AUC) values of bSIRT2, AFC, AMH, and age were significantly different from those under the opportunity reference line (P<0.05). In the subsequent correlation analysis, FFSIRT2, and HCG day serum SIRT2 were negatively correlated with age (r=-0.35, r=-0.19), and positively correlated with AFC (r=0.2, r=0.02). Serum SIRT1 on HCG day was negatively correlated with the number of blastocysts and the number of frozen embryos (r=-0.18, r=-0.21). Levels of FF SIRT1 and FF SIRT2 were significantly lower than those in serum SIRT1 and SIRT2, and there was no significant difference in serum SIRT1 and SIRT2 before and after ovulation promotion. CONCLUSIONS: The results suggest that bSIRT2 has significant statistical significance in predicting the cumulative number of pregnancies. When combined with AMH, AFC, and age, bSIRT2 can predict the cumulative pregnancy outcome. In addition, the level of serum SIRT1 and SIRT2 were not affected by ovulation promotion.

Comparative study of assisted reproductive outcomes between young patients with occult premature ovarian insufficiency and advanced-age patients.

OBJECTIVE: The purpose of this study was to compare the pregnancy outcomes among young patients with occult premature ovarian insufficiency (OPOI), advanced-age patients with diminished ovarian reserve (DOR), and advanced-age patients with normal ovarian reserve. METHODS: We retrospectively reviewed 324 women who underwent their first cycles of in vitro fertilization/intracytoplasmic sperm injection. The women were divided into the following groups: young women with OPOI, advanced-age women with DOR, and advanced-age women with normal ovarian reserve. The outcomes were compared among the different groups. RESULTS: The rates of live birth and embryo implantation in the young OPOI group were significantly higher than in the advanced-age DOR group, but comparable to those in the advanced-age normal ovarian reserve group. Moreover, the abortion rate was significantly lower in young OPOI patients compared with advanced-age patients with or without DOR. CONCLUSION: Higher embryo implantation and live birth rates and a lower abortion rate can be achieved in young patients with OPOI compared with older patients. The better outcomes in advanced-age patients with normal ovarian reserve compared with DOR may be related to egg quantity rather than quality.

P53 Plays an Important Role in the Early Stage of Autologous Heterotopic Transplantation of Ovaries into the Backs of Mice.

BACKGROUND: Subcutaneous ovarian transplantation has recently begun receiving increased attention. Fourteen days after transplantation is used as an important time point for assessing the recovery of ovarian function. The goal of this study is to determine the expression of apoptotic genes in the ovary at this time. METHODS: This study investigated follicle development and the expression of 3 apoptosis genes (Bax, Bcl2, and P53) after mouse ovaries were transplanted. Seven-week-old mouse ovaries were autologously transplanted into back muscle. The ovaries were harvested on day 14, morphology was observed by hematoxylin and eosin staining, and the distribution of 3 proteins was observed by immunohistochemistry. TUNEL staining showed where apoptosis occurred in the ovary. Finally, RT-PCR/Western blotting was used to analyze the differential expression of mRNA/proteins between the transplantation group and the control group. RESULTS: The results revealed follicles at different stages at the edge of the grafts. In immunohistochemical experiments, BAX, BCL2, and P53 were found to be extensively expressed in the transplant group and the control group. P53 was strongly expressed in the medulla of transplanted ovaries. Bax was strongly expressed in the antral follicles of both groups. The results were consistent with the results of the TUNEL experiments. Three genes (Bax, Bcl2, and P53) were downregulated in the transplanted groups. The results showed that significant differences were detected in Bax and P53 mRNA expression levels between the transplanted groups and the control group (P < .01). Bcl2 expression was not significantly different, but the Bax/Bcl2 ratio increased. The results of the protein experiments were the same. CONCLUSION: P53 may downregulate Bax in the early stage of transplantation. Follicle growth and atresia were regulated through modulation of Bcl2- and Bax-mediated apoptotic pathways in heterotopic ovarian transplantation.

Protective effects of l-carnitine on astheno- and normozoospermic human semen samples during cryopreservation.

This study was conducted to determine the effects of l-carnitine (LC), as an antioxidant, in preventing spermatozoa damage during the freezing-thawing process in both astheno- and normozoospermic human semen samples. Seventy semen samples (37 asthenozoospermic and 33 normozoospermic) were involved in this study. Cryopreservation medium supplemented with 1.0 g/l LC was mixed with semen at a ratio of 1:1 (v/v). Controls were cryopreserved with freezing medium only. Assessment of motility, viability (VIA), mitochondrial membrane potential (MMP) and DNA fragmentation index (DFI) were performed on aliquots of fresh semen, frozen-thawed control and frozen-thawed LC treated samples. Supplementation of the cryopreservation medium with LC induced a significant improvement in post-thaw sperm parameters in both the asthenozoospermic and normozoospermic semen samples, compared with those of the control, regarding sperm fast forward motility, forward motility, total motility and VIA. LC showed better protective effects towards asthenozoospermia for DFI (F = 115.85, P < 0.01) and VIA (F = 67.14, P < 0.01) than did normozoospermic semen samples. We conclude that supplementation with LC prior to the cryopreservation process reduced spermatozoa cryodamage in both asthenozoospermic and normozoospermic semen samples. LC had better protective effects for asthenozoospermic human semen samples. Future research should focus on the molecular mechanism for and the different protective effects of LC between asthenozoospermic and normozoospermic semen samples during cryopreservation.

The role of serum AMH and FF AMH in predicting pregnancy outcome in the fresh cycle of IVF/ICSI: a meta-analysis.

PURPOSE: This meta-analysis tries to find and confirm the true prognostic value of serum AMH and the follicle fluid AMH (FF AMH) on the outcome of ART. METHOD: We identified all studies published by March 2014 with data related to in vitro fertilization", "intracytoplasmic sperm injection", "assisted reproductive technology" and "antimullerian hormone" in Pubmed database. Studies were included if 2 × 2 tables for outcomes of pregnancy in IVF patients in relation to AMH could be constructed or studies which used T-tests to compare clinical indexes including AMH in pregnant and non-pregnant women. And all the patients were less than 46 years old. RESULTS: A total of 26 studies could be used for this meta-analysis. Of these articles 22 studies could be constructed 2 × 2 tables, with 15 for predicting pregnancy and 7 for non-pregnancy. 11 studies used the analysis of T-test, with 7 articles were duplicated. And of the 11 articles, 8 were for the analysis of serum AMH in prediction of pregnancy, 3 were for FF AMH. Because of heterogeneity among studies, calculation of a summary point estimate for sensitivity and specificity was not possible. For the analysis of serum AMH on non-pregnancy, the heterogeneity was moderate (I-squared of 65.9%), the curves indicated positive find (the AUROC is 0.73, 95% CI is 0.69-0.77.). In the T-test group of serum AMH, the DOR for women with pregnancy outcome was 0.232 (95% confidence interval (CI): 0.034-0.43), with less heterogeneity (I-squared of 45.1%). Unfortunately, the predictive value of FF AMH on pregnancy is still unclear because of large heterogeneity (I-squared of 90.5%). CONCLUSION: Serum AMH, as an independent parameter, can predict pregnancy outcome after assisted conception and the positive correlation with serum AMH and non-pregnancy should not be ignored either. The predictive value of FF AMH on pregnancy is still unclear.

The predictive value of anti-Mullerian hormone on embryo quality, blastocyst development, and pregnancy rate following in vitro fertilization-embryo transfer (IVF-ET).

PURPOSE: The objective of this study was to investigate the predictive value of anti-Mullerian hormone (AMH) on fertilization rate (FR), blastocyst development, embryo quality, the outcome of the pregnancy and the live birth rate (LBR) following in vitro fertilization-embryo transfer (IVF-ET)/intracytoplasmic sperm injection (ICSI). METHOD: In this prospective study outcomes were followed in 83 women undergoing cycles of IVF/ICSI within a university hospital. Basal serum AMH, follicle stimulating hormone (FSH), luteinizing hormone (LH) and antral follicle count (AFC) were measured on Day 3. Serum AMH (Gn6 AMH ) level was measured on Day 6 after the administration of gonadotrophin (Gn). AMH was measured in follicle fluid (FF AMH) on the day of ovum pick-up (dOPU). The numbers of retrieved and fertilized oocytes, good quality embryos and blastocysts were counted. Secondary outcome variables included clinical pregnancy rate (CPR) and LBR. RESULTS: Spearman correlation analysis indicated that the numbers of oocytes, good quality embryos and blastocysts were associated with AMH (P < 0.05) and that LBR was correlated with FF AMH (r = 0.495, P < 0.05). No associations were found between FR and AMH (P > 0.05). Receiver operating characteristic analysis showed that the sensitivity of FF AMH at predicting CPR was 91.2%; the specificity was 86.5% and ROC(AUC) was 0.893 (P < 0.0001). CONCLUSION: AMH parameters were correlated with good quality embryos and blastocysts, but only FF AMH showed a significant correlation with LBR and CPR.