Effect of follicle size on pregnancy outcomes in patients undergoing first letrozole-intrauterine insemination.

BACKGROUND: Letrozole has been proven to be an effective method for inducing ovulation. However, little attention has been paid to whether the lead follicle size will affect the success rate of intrauterine insemination (IUI) with ovulation induction with alone letrozole. Therefore, we hope to investigate the effect of dominant follicle size on pregnancy outcomes on human chorionic gonadotropin (hCG) day of the first letrozole-IUI. METHODS: A retrospective cohort study design was employed. We included patients with anovulation or unexplained infertility undergoing first IUI treatment with letrozole for ovarian stimulation. According to the dominant follicle size measured on the day of hCG trigger, patients were divided into six groups (≤ 18 mm, 18.1-19.0 mm, 19.1-20.0 mm, 20.1-21.0 mm, 21.1-22.0 mm, > 22 mm). Logistic models were used for estimating the odds ratios (ORs) with their 95% confidence interval (CIs) for achieving a clinical pregnancy or a live birth. A restricted cubic spline was drawn to explore the nonlinear relationship between follicle size and IUI outcomes. RESULTS: A total of 763 patients underwent first letrozole-IUI cycles in our study. Fisher exact test showed significant differences among the six follicle-size groups in the rates of pregnancy, clinical pregnancy and live birth (P < 0.05 in each group). After adjusting the potential confounding factors, compared with the follicles ≤ 18 mm in diameter group, 19.1-20.0 mm, 20.1-21.0 mm groups were 2.3 or 2.56 times more likely to get live birth [adjusted OR = 2.34, 95%CI (1.25-4.39); adjusted OR = 2.56, 95% CI (1.30-5.06)]. A restricted cubic spline showed an inverted U-shaped relationship between the size of dominant follicles and pregnancy rate, clinical pregnancy rate, and live birth rate, and the optimal follicle size range on the day of hCG trigger was 19.1-21.0 mm. When the E2 level on the day of hCG trigger was low than 200 pg/mL, the clinical pregnancy rates of 19.1-20.0 mm, 20.1-21.0 mm groups were still the highest. CONCLUSIONS: The optimal dominant follicle size was between 19.1 and 21.0 mm in hCG-triggered letrozole-IUI cycles. Either too large or too small follicles may lead to a decrease in pregnancy rate. Using follicle size as a predicator of pregnancy outcomes is more meaningful when estrogen on the day of hCG trigger is less than 200 pg/ml.

Hyperlipidemia is negatively associated with pregnancy outcomes in patients following frozen-thawed embryo transfer: A retrospective study.

Background and Aims: It is demonstrated that lipid metabolism disorders are associated with the reproductive performances of assisted reproductive technology. However, it is little known whether hyperlipidemia is associated with the endometrial receptivity and pregnancy outcomes of patients undergoing frozen-thawed embryo transfer (FET). Methods: This was a retrospective analysis involving 554 infertile women undergoing FET. The patients were divided into the hyperlipidemia group (n = 224) and control group (n = 320) based on the levels of serum lipids. The clinical and laboratory indexes between the two groups were compared. Meanwhile, the stratified analysis based on body mass index (BMI) and endometrial preparation protocols was performed. The independent samples t-test, Mann-Whitney U test, χ2 test and multiple logistic regression analysis were used to compare and analyze the data. Results: The patients with hyperlipidemia had significantly higher serum lipids levels and BMI and lower clinical pregnancy and implantation rates than those with normal blood lipids (p < 0.05). The impact of hyperlipidemia on pregnancy outcomes was independent of BMI. The multiple logistic regression analysis showed that higher cholesterol was associated with lower pregnancy rate and implantation rate (p < 0.05). Regardless of blood lipid levels, the patients undergoing the hormone replacement therapy (HRT) protocol had higher estradiol levels and lower progesterone levels compared with the stimulated cycles (STC) (p < 0.05). Moreover, the clinical pregnancy rate and implantation rate of the HRT protocol were higher than those of the STC, although there was no significant difference between the two. Conclusion: Hyperlipidemia especially higher cholesterol has a negative effect on the pregnancy outcomes of the patients undergoing FET. Actively implementing lipid-lowering treatment and the HRT protocol seem more friendly for these patients.

Effect of interval time between hysterosalpingography and intrauterine insemination on the pregnancy outcome of infertile patients.

Background: Hysterosalpingography (HSG) is the most commonly applied tubal patency test in clinical practice. Although some studies have found an increased pregnancy rate after HSG, no studies to date have specifically characterized the effect of interval time between HSG and IUI on pregnancy outcome. Objectives: To investigate the effect of interval time between HSG and intrauterine insemination (IUI) on live birth rates of infertile patients. Methods: Retrospective cohort study. The reproductive medical record system was used to identify patients who completed ≥1 IUI cycle between January 2017 and October 2021. According to the interval time between HSG and IUI, patients were divided into three groups: <6months interval group,6-12 months interval group and >12 months interval group. The generalized estimating equation with Poisson distribution was used to estimate the risk ratios (RRs) and 95% confidence intervals (CIs) of different groups. Results: A total of 413 patients completed 701 IUI cycles during the study period, <6months interval group, 415 cycles; 6-12 months interval group, 138 cycles; >12 months interval group, 148 cycles. The live birth rate of <6 months group was higher than other two groups (17.35% vs. 12.32% vs. 8.11%, P=0.017); Similarly, the clinical pregnancy rate of <6 months group was also higher than other two groups (19.76% vs. 14.49% vs.11.49%, P=0.049). When adjusted separately for FSH, AMH, infertility type, duration of infertility, infertility diagnosis, total motile count (TMC) of sperm, medications, endometrium size and dominant follicle size, the live birth rate of >12 months group severally significantly decreased by 60% (adjusted RR = 0.40, 95% CI [0.19-1.40]). The cumulative clinical pregnancy and live birth rates of <6 months group were higher than other two groups (P<0.05), but the cumulative pregnancy rate among three groups were not statistically different (log rank test: P=0.06). Conclusion: The interval time between hysterosalpingography and IUI is related to pregnancy outcome. The clinical pregnancy and live birth rates were the highest when the time interval was less than 6 months. Therefore, IUI should be recommend as soon as possible after HSG if the patient couple meets the IUI indication.

Performance evaluation of sperm concentration, motility, and morphological analysis for GSA-810 series of sperm quality analysis system.

BACKGROUND: The performance evaluation of each computer-assisted sperm analysis (CASA) system may provide a basis for the interpretation of clinical results and further improvement of the CASA system. METHODS: The accuracy of the GSA-810 CASA system was evaluated by detecting latex bead quality control products. The precision of sperm concentration, morphology, and percentages of progressively motile sperm (PR) were evaluated by coefficient of variation (CV). Three samples with sperm concentration of about 100 × 106 /mL were diluted to evaluate the linear range. RESULTS: The detection values of latex beads were within the range of target values. The CVs of sperm concentration and PR were significantly and negatively correlated with sperm concentration (r = -0.561, p = 0.001) and PR value (r = -0.621, p < 0.001), respectively. The R2 values of the linear range of sperm concentration were ≥0.99. There was no significant difference in sperm motility and PR within 1-10 min at 36.5°C ± 0.5°C. The coincidence rates of sperm morphology and sperm head morphology for 36 semen samples analyzed by the GSA-810 system and manual method were 99.40% and 99.67%, respectively. The CVs of the percentage of sperm with abnormal morphology and percentage of sperm with abnormal head morphology were less than 5%. CONCLUSION: The GSA-810 system can accurately analyze normal semen samples, but the repeatability of the results is poor for oligozoospermia and asthenozoospermia samples. The future CASA system for analyzing sperm morphology should focus on recognizing the middle and tail segments of a spermatozoon.

DTLR-CS: Deep tensor low rank channel cross fusion neural network for reproductive cell segmentation.

In recent years, with the development of deep learning technology, deep neural networks have been widely used in the field of medical image segmentation. U-shaped Network(U-Net) is a segmentation network proposed for medical images based on full-convolution and is gradually becoming the most commonly used segmentation architecture in the medical field. The encoder of U-Net is mainly used to capture the context information in the image, which plays an important role in the performance of the semantic segmentation algorithm. However, it is unstable for U-Net with simple skip connection to perform unstably in global multi-scale modelling, and it is prone to semantic gaps in feature fusion. Inspired by this, in this work, we propose a Deep Tensor Low Rank Channel Cross Fusion Neural Network (DTLR-CS) to replace the simple skip connection in U-Net. To avoid space compression and to solve the high rank problem, we designed a tensor low-ranking module to generate a large number of low-rank tensors containing context features. To reduce semantic differences, we introduced a cross-fusion connection module, which consists of a channel cross-fusion sub-module and a feature connection sub-module. Based on the proposed network, experiments have shown that our network has accurate cell segmentation performance.

Effect of hyperlipidemia on the outcome of in vitro fertilization in non-obese patients with polycystic ovary syndrome.

Introduction: It is little known whether hyperlipidemia alone has adverse effects on the outcome of in vitro fertilization (IVF) in patients with polycystic ovarian syndrome (PCOS). Methods: The PCOS patients with body mass index (BMI) < 30 kg/m2 were performed IVF or intracytoplasmic sperm injection treatment, including 208 fresh cycles and 127 frozen embryo transfer (FET) cycles. All the patients were divided into hyperlipidemia and control groups, and embryo quality and pregnancy outcomes between the two groups were compared. Results: In the fresh cycles, total gonadotropin dosage in the control group was significantly lower than that in the hyperlipidemia group, and serum estradiol levels on trigger day were reversed (P < 0.05). The embryo fragment score was positively correlated with serum low-density lipoprotein level (r = 0.06, P < 0.05) and negatively with serum high-density lipoprotein (HDL) and lipoprotein A levels (r = -0.489 and -0.085, P < 0.01). Logistic regression analysis found that HDL was beneficial for clinical pregnancy (OR = 0.355, 95% CI: 0.135-0.938, P < 0.05). In the FET cycles, there were no differences in pulse index, systolic/diastolic ratio and serum estradiol and progesterone levels between the two groups, but resistance index in the hyperlipidemia group was significantly higher than that in the control group (P < 0.05). Conclusion: Hyperlipidemia may increase the dosage of gonadotropin and have adverse effect on the embryo quality, endometrial receptivity, and clinical outcomes of lean PCOS patients. It is recommended that the non-obese patients with hyperlipidemia and PCOS perform lipid-lowering treatment before undergoing embryo transfer.

Association between gut microbiota and benign prostatic hyperplasia: a two-sample mendelian randomization study.

Background: Recent researches have shown a correlation between the gut microbiota (GM) and various diseases. However, it remains uncertain whether the relationship between GM and benign prostatic hyperplasia (BPH) is causal. Methods: We carried out a two-sample Mendelian randomization (MR) analysis, utilizing data from the most extensive GM-focused genome-wide association study by the MiBioGen consortium, with a sample size of 13,266. Data for BPH, encompassing 26,358 cases and 110,070 controls, were obtained from the R8 release of the FinnGen consortium. We employed multiple techniques, such as inverse variance weighted (IVW), constrained maximum likelihood and model averaging methods, maximum likelihood, MR-Pleiotropy RESidual Sum and Outlier (MRPRESSO),MR-Egger, and weighted median methods, to investigate the causal relationship between GM and BPH. To evaluate the heterogeneity among the instrumental variables, Cochran's Q statistics were employed. Additionally, the presence of horizontal pleiotropy was assessed through the application of both MR-Egger and MR-PRESSO tests. The direction of causality was scrutinized for robustness using the MR-Steiger directionality test. A reverse MR analysis examined the GM previously linked to BPH through a causal relationship in the forward MR assessment. Results: According to the analysis conducted using IVW,Eisenbergiella (odds ratio [OR]=0.92, 95% confidence interval [CI]: 0.85-0.99,P=0.022) and Ruminococcaceae (UCG009) (OR=0.88, 95% CI: 0.79-0.99, P=0.027) were found to reduce the risk of BPH, while Escherichia shigella (OR=1.19, 95% CI: 1.05-1.36, P=0.0082) appeared to increase it. The subsequent reverse MR analysis revealed that the three GM were not significantly influenced by BPH, and there was no noticeable heterogeneity or horizontal pleiotropy among the instrumental variables.Conclusion: These results indicated a causal relationship between Eisenbergiella, Ruminococcaceae (UCG009), and Escherichia shigella and BPH. Further randomized controlled trials are needed to explore more comprehensively the roles and operational mechanisms of these GM in relation to BPH.

The causal association between systemic inflammatory regulators and primary ovarian insufficiency: a bidirectional mendelian randomization study.

BACKGROUND: Recent studies have suggested a potential link between systemic inflammatory regulators and primary ovarian insufficiency (POI); however, a causal relationship between them remains unclear. In this study, we explored the causal link between systemic inflammatory regulators and POI risk using a bidirectional, two-sample Mendelian randomization (MR) strategy. RESULTS: This approach utilized the most extensive genome-wide association study involving 41 systemic inflammatory regulators in a sample of 8,293 Finnish individuals and POI data from the FinnGen consortium (254 cases vs. 118,228 controls). The inverse variance weighting approach served as a primary MR method, and four additional MR techniques (Maximum Likelihood, MR-Egger, Weighted Median, and constrained maximum likelihood and model averaging Bayesian information criterion ) were applied to support and validate results. Cochran's Q statistics were used to assess the heterogeneity of instrumental variables, whereas the MR-Egger and MR Pleiotropy Residual Sum and Outlier tests detected horizontal pleiotropy. The MR Steiger test evaluated the strength of a causal association. Our findings suggest that lower levels of vascular endothelial growth factor (odds ratio [OR] = 0.73, 95% confidence interval [CI]: 0.54-0.99, P = 0.046) and interleukin-10 (OR = 0.54, 95% CI: 0.33-0.85, P = 0.021) are associated with an increased risk of POI. Reverse MR analysis revealed no significant effect of POI on the expression of these 41 systemic inflammatory regulators. No notable heterogeneity or horizontal pleiotropy was observed in the instrumental variables. CONCLUSIONS: This study revealed a causal association between 41 systemic inflammatory regulators and POI, demonstrating that decreased levels of VEGF and IL-10 are linked to an elevated risk of POI. Further investigations are necessary to assess the potential of these biomarkers as early predictors, preventive strategies, and therapeutic targets for POI.

Association between gut microbiota and primary ovarian insufficiency: a bidirectional two-sample Mendelian randomization study.

Background: Recent studies have indicated a potential correlation between intestinal bacteria and primary ovarian insufficiency (POI). However, the causal relationship between the gut microbiota (GM) and POI remains unclear. Methods: A bidirectional two-sample Mendelian randomization (MR) study was conducted to investigate the relationship between the GM and POI. Data on the GM were based on the MiBioGen consortium's summary statistics from the most comprehensive genome-wide association study meta-analysis to date (n=13,266), and POI data were obtained from the R8 release of the FinnGen consortium, containing a total of 424 cases and 181,796 controls. A variety of analytical methods, including inverse variance weighting, maximum likelihood, MR-Egger, weighted median, and constrained maximum likelihood and model averaging and Bayesian information criterion, were utilized to explore the connection between the GM and POI. The Cochran's Q statistics were used to evaluate the heterogeneity of instrumental variables. The MR-Egger and MR-pleiotropy residual sum and outlier (PRESSO) methods were used to identify the horizontal pleiotropy of instrumental variables. The MR Steiger test was used to evaluate the strength of causal relationships. A reverse MR study was performed to investigate the causal relationship between POI and the targeted GMs which were indicated to have a causal relationship with POI in the forward MR evaluation. Results: The inverse variance weighted analysis indicated that Eubacterium (hallii group) (odds ratio [OR]=0.49, 95% confidence interval [CI]: 0.26-0.9, P=0.022) and Eubacterium (ventriosum group) (OR=0.51, 95% CI: 0.27-0.97, P=0.04) had protective effects on POI, and Intestinibacter (OR=1.82, 95% CI: 1.04-3.2, P=0.037) and Terrisporobacter (OR=2.47, 95% CI: 1.14-5.36, P=0.022) had detrimental effects on POI. Results of the reverse MR analysis indicated that POI had no significant influence on the four GMs. No significant heterogeneity or horizontal pleiotropy was observed in the performance of the instrumental variables. Conclusion: This bidirectional two-sample MR study revealed a causal link between Eubacterium (hallii group), Eubacterium (ventriosum group), Intestinibacter, and Terrisporobacter and POI. Additional clinical trials are needed to gain a clearer understanding of the beneficial or detrimental effects of the GMs on POI and their mechanisms of action.

Analysis of selected sperm samples by a computer-assisted system with high frame rate: A prospective study.

Background and Aims: Due to the rapid motility of the selected sperm, sperm parameters cannot be accurately determined by the manual method. So, the application of a computer-assisted sperm analysis system with a high frame rate (HFR-CASA, 85 Hz) in sperm selection is investigated. Methods: A total of 177 semen samples were collected for sperm selection with density gradient centrifugation. Then, the manual method and HFR-CASA method will be used to observe and analyze the sperm concentration, motility, and percentage of progressively motile sperm (PR) of the selected sperm samples. The quality control of sperm concentration was performed with microballoons. Two selected sperm samples were analyzed 10 times repeatedly to evaluate the accuracy of the HFR-CASA. Results: The results of microballoons analyzed with the HFR-CASA were in control. The coefficients of variation of sperm concentration, motility, and PR from two selected sperm samples were all below 10%. The results of 177 selected sperm samples analyzed by the manual method and HFR-CASA showed that although there were significant positive correlations in sperm concentration, motility, and PR between them (p < 0.001), the manual method significantly underestimated sperm concentration (p = 0.002) but overestimated sperm motility and PR (p < 0.001). When sperm concentration was below 50 × 106/mL, the manual method significantly overestimated sperm concentration (p < 0.05). However, when sperm concentration was more than 100 × 106/mL, the manual method significantly underestimated sperm concentration (p < 0.001). Regardless of sperm concentration, the manual method consistently overestimated sperm motility and PR (p < 0.001). When sperm concentration was more than 20 × 106/mL, there was no correlation in sperm PR between them (p > 0.05). When sperm concentration was below 50 × 106/mL, the correct rate of captured sperm by the HFR-CASA was more than 98%. Conclusion: The HFR-CASA method is more accurate than the manual method in analyzing the selected sperm samples.

Correlations of joint detection of 22 vaginal microbes with routine examination results of vaginal secretions and assisted reproductive outcomes.

The correlations of joint detection of 22 vaginal microbes with routine examination results of vaginal secretions and assisted reproductive outcomes were investigated. There were 37 samples with abnormal vaginal microecology in 107 vaginal secretion samples. The top 5 detection rates of microorganisms were Ureaplasma urealyticum (73.83%), Prevotella sp. (70.09%), Gardnerella vaginalis (53.27%), L. crispatus (52.34%) and L. inerts (51.40%). When the levels of Bacillus and hydrogen peroxide in vaginal secretions decreased or pH increased, the abnormal rates of vaginal microecology increased significantly (P < 0.01). The clinical pregnancy rate (53.66%, 22/41) in the women with normal vaginal microecology was higher than that (37.5%, 9/24) with abnormal vaginal microecology. In conclusions, the joint detection of 22 vaginal microbes can quickly and effectively determine whether the vaginal microecology is normal or not. The evaluation of vaginal microecology may be valuable in predicting the assisted reproductive outcomes of infertile women.

Personalized versus standard frozen-thawed embryo transfer in IVF/ICSI cycles: a systematic review and meta-analysis.

PURPOSE: To investigate whether personalized embryo transfer (pET) protocol guided by an endometrial receptivity array (ERA) can improve clinical outcomes of assisted reproduction. METHODS: We searched PubMed, Embase, Web of Science, and the Cochrane library for studies in which analytical comparisons of outcomes of pET and standard embryo transfer (sET) groups were undertaken. The references to the included studies were also manually searched. The primary outcome was clinical pregnancy rate (CPR), and the secondary outcomes were live birth rate (LBR), human chorionic gonadotropin (HCG) positivity, biochemical pregnancy rate (BPR), miscarriage rate (MR), implantation rate (IR), and ongoing pregnancy rate (OPR). RESULTS: Ten studies were included in the meta-analysis, including one randomized controlled trial (RCT) and nine cohort studies. We observed no significant difference in the primary outcome of CPR between the pET and sET groups in unselected patients (RR = 1.07; 95% confidence interval [CI], 0.87-1.30; P = 0.53; I2 = 89%). In terms of secondary outcomes, we likewise noted no significant differences between the groups. Further subgroup analyses indicated that the pET protocol not only significantly reduced the MR for poor-prognosis patients, but it also reduced the CPR in donor cycles, elevated the BPR for good-prognosis patients, non-preimplantation genetic testing (PGT), and programmed cycles, and decreased the proportion of women showing HCG positivity in non-PGT cycles. CONCLUSIONS: This meta-analysis revealed that ERA appears to possess limited guidance in embryo transfer. More high-quality RCTs are therefore needed to investigate the clinical validity and feasibility of ERA in the future.

Assisted biomimetic electrostimulation therapy can improve the clinical pregnancy rate of patients with abnormal endometrial receptivity undergoing frozen-thawed embryo transfer cycles.

This study aimed to explore whether assisted biomimetic electrostimulation (BES) therapy can improve clinical outcomes in patients with abnormal endometrial receptivity undergoing frozen-thawed embryo transfer (FET) cycles. We retrospectively collected data from 132 patients who underwent FET cycles and divided them into the BES (n = 86) and non-BES (NBES) groups (n = 46). The clinical pregnancy rate (55.8 vs. 37.0%), biochemical pregnancy rate (59.3 vs. 41.3%), and live birth rate (44.2 vs. 23.9%) of the BES group were significantly higher than those of the NBES group (p < 0.05). No significant difference between the two groups was observed in endometrial thickness at FET day, embryo implantation rate, and early abortion rate (p > 0.05). The logistic regression analysis indicated that blastocyst transfer (adjusted OR = 3.617; 1.337-9.783; p = 0.011) and BES (adjusted OR = 2.398; 1.094-5.256; p = 0.029) were positively associated with the clinical pregnancy rate. These results suggested that assisted BES therapy can improve clinical outcomes in patients with diseases affecting endometrial receptivity.Impact statementWhat is already known on this subject? Biomimetic electrostimulation (BES) therapy can increase endometrial thickness in patients with thin endometria undergoing embryo transfers and to some extent improve their clinical outcomes.What do the results of this study add? Assisted BES therapy can improve clinical pregnancy rates in patients with abnormal endometrial receptivity undergoing FET cycles (55.8 vs. 37.0%, p = 0.039). After adjusting for covariates, BES was still positively associated with the clinical pregnancy rate (adjusted OR = 2.398; 1.094-5.256; p = 0.029).What are the implications of these findings for clinical practice and/or further research? BES therapy can improve endometrial receptivity. Further studies are needed to understand its specific mechanisms.

Screening of sperm antigen epitopes by phage display technique and its preliminary clinical application.

BACKGROUND: At present, there is a lack of standardized preparation methods of sperm antigen for the detection of antisperm antibody (AsAb). To screen sperm antigen mimotopes from a phage display random peptide library and use them to establish an enzyme-linked immunosorbent assay (ELISA) for the detection of AsAb, immunoglobulins were extracted from the sera of rabbits with positive AsAb and negative AsAb, respectively, by the saturated ammonium sulfate method, and a phage display 12-mer peptide library was affinity panned by the extracted immunoglobins coated on the ELISA plate. Then, the obtained positive phage clones were identified by ELISA and sent for sequencing and peptides synthesis. Last, a diagnostic ELISA was established to detect clinical serum and seminal plasma samples. RESULTS: A total of sixty phage clones were chosen by affinity panning, and sixteen of them reacted positively with AsAb in indirect ELISA and sandwich ELISA. Following DNA sequencing and translation, the peptide sequences of the sixteen positive clones were obtained. By comparison in Blast database, four of sixteen positive clones were found to be closely related to male reproduction. Two (#1 and #25) of four mimotopes were synthesized, and an ELISA method was established using the two mimotopes as sperm specific antigens. One hundred and thirty-four serum samples and seventy-four seminal plasma samples from infertile couples were analyzed by the established ELISA with #1 and #25 mimotopes, respectively. The positive rates of AsAb in serum samples were 20.15% (27/134) for #1 and 11.19% (15/134) for #25, respectively, and the coincidence rate between them was 91.04% (122/134). The positive rates of AsAb in seminal plasma samples were 1.35% (1/74) for both #1 and #25, and the coincidence rate was 100%. CONCLUSION: Sperm antigen mimotopes can be obtained successfully by the phage display technique, and can be used as standard sperm specific antigens to establish an ELISA method for the detection of AsAb.

RéSUMé: CONTEXTE: À ce jour, il n'existe pas de méthodes normalisées de préparation d'antigènes spermatiques pour la détection des anticorps anti-spermatozoïdes (ACAS). Dans le but d'élaborer un tel test ELISA (enzyme-linked immunosorbent assay), nous avons extrait de sérum de lapins des anticorps anti-spermatozoïdes humains via la technique du sulfate d'ammonium saturé et en ayant recours à une librairie phagique de peptides (12-mer). Les clones positifs ont été identifiés par ELISA, séquencés à façon et les peptides correspondants ont été synthétisés. In fine, un test ELISA diagnostic a été conçu pour être utilisé avec des échantillons cliniques de sérum et de plasmas séminaux. RéSULTATS: Au total, soixante clones de phages ont été sélectionnés, et seize d'entre eux se sont avérés interagir avec les ACAS en ELISA indirect comme en ELISA sandwich. Les séquences peptidiques de ces seize clones positifs ont été obtenues. Par comparaison avec les bases de données (Blast), quatre de ces seize clones positifs se sont révélés être étroitement liés à la reproduction masculine. Deux des quatre mimotopes (#1 et #25) ont été synthétisés, et un test ELISA a été généré en utilisant ces deux mimotopes comme antigènes spécifiques des spermatozoïdes. Cent trente-quatre échantillons de sérum et soixante-quatorze échantillons de plasma séminal de patients de couples infertiles ont alors été analysés avec ce test ELISA. Respectivement, les échantillons sériques se sont révélés positifs à 20,15% (27/134) pour le mimotope #1 et à 11,19% (15/134) pour le mimotope #25, avec un taux de coïncidence de 91,04% (122/134). Seul un échantillon de plasma séminal (1/74, soit 1, 35%) s'est révélé positif à la fois pour le mimotope #1 et #25 (coïncidence 100%). CONCLUSION: La technique « phage display¼ nous a permis d'identifier des mimotopes d'antigènes spermatiques qui ont pu être utilisés afin de générer un test ELISA pour la détection d'anticorps anti-spermatozoïdes.

CircRNA circFADS2 is Downregulated in Endometritis and its Overexpression Promotes miR-643 Maturation in Human Endometrial Epithelial Cells to Suppress Cell Apoptosis.

CircRNA circFADS2 suppresses LPS-induced inflammation, which plays a critical role in endometritis. Our preliminary sequencing analysis revealed a positive correlation between circFADS2 and miR-643, which also play protective roles in LPS-induced inflammation. Therefore, this study was performed to explore the involvement of circFADS2 in endometritis with a focus on its interaction with miR-643. RT-qPCR was performed to analyze the levels circFADS2, mature miR-643, and premature miR-643 in plasma samples from endometritis patients (n = 66) and healthy controls (n = 66). Pearson's correlation coefficient was applied to analyze correlations between these genes. The effect of circFADS2 on miR-643 maturation was analyzed by measuring miR-643 and premature miR-643 levels in circFADS2-overexpressed human endometrial epithelial cell line HEnEpCs. The role of circFADS2 and miR-643 in HEnEpC apoptosis under LPS treatment was analyzed by cell apoptosis assay. CircFADS2 was downregulated in endometritis and was positively correlated with mature miR-643, but not premature miR-643. CircFADS2 overexpression in HEnEpCs increased the level of mature miR-643 but not premature miR-643. Cell apoptosis analysis showed that circFADS2 and miR-643 overexpression protected HEnEpCs from LPS-induced cell apoptosis, and miR-643 inhibition reduced the effect of circFADS2 overexpression. CircFADS2 is downregulated in endometritis, and it overexpression promotes miR-643 maturation in HEnEpCs to suppress cell apoptosis.

Gut microbiota dysbiosis in polycystic ovary syndrome: association with obesity - a preliminary report.

The objective was to explore if and how the microbiota changed in polycystic ovary syndrome (PCOS) women compared with healthy women. Eight obese PCOS (PO group), 10 nonobese PCOS (PN group), and nine healthy normal weight women (control) (C group) were enrolled. Insulin (INS), testosterone (T), follicle-stimulating hormone (FSH), luteinizing hormone (LH), estrogen (E2), and dehydroepiandrosterone (DHEA) were detected with radioimmunoassay. Antimullerian hormone (AMH), fasting glucose, and hemoglobin A1c (HbA1c) were determined by a chemiluminescence immunoassay, glucose oxidase method, and HPLC, respectively. Gut microbiota composition was evaluated by PCR. Alpha diversity was assessed using Chao1 and the Shannon index. PCOS women showed significantly higher T, LH, and LH/FSH and lower FSH levels than the C group (p < 0.05). The AMH level was significantly higher in the PO than in the PN group (p < 0.05). The PO group presented a significantly higher fasting INS level and HMOA-IR scores than the other groups, lower observed SVs and alpha diversity than the C group, higher beta diversity than the PN group (p < 0.05), and decreased abundances of genera (mainly butyrate producers). Regression analysis showed that decreased abundances of several genera were correlated with higher circulating T and impaired glucose metabolism. PCOS is associated with changes in the gut microbiota composition. Obesity has a driving role in the development of dysbiotic gut microbiota in PCOS.

Alpinetin ameliorates inflammatory response in LPS-induced endometritis in mice.

Alpinetin has been reported to have anti-inflammatory effects. However, whether alpinetin has anti-inflammatory effects in LPS-induced endometritis has not been thoroughly elucidated to date. The aim of this study was to investigate the protective effects of alpinetin on LPS-induced endometritis in mice. A mouse model of endometritis was induced by LPS and alpinetin was given 1 h before LPS treatment. According to the results, alpinetin protected mice against LPS-induced endometritis by attenuating uterine histological changes and myeloperoxidase (MPO) activity. The LPS-induced inflammatory response was inhibited by alpinetin as confirmed by the inhibition of TNF-α, IL-1ß, and IL-6 production. Furthermore, LPS-induced TLR4 expression and NF-κB activation were significantly suppressed by alpinetin. In addition, the expression of PPAR-γ was dose-dependently increased by the treatment of alpinetin. Taken together, the results of this study showed that alpinetin had protective effects against LPS-induced endometritis in mice, and the beneficial effects were occurred through the activation of PPAR-γ and inhibition of the TLR4 signaling pathway.

Analysis of human sperm DNA fragmentation index (DFI) related factors: a report of 1010 subfertile men in China.

BACKGROUND: Many factors may lead to sperm DNA damage. However, it is little known that the correlations of sperm DNA damage with obesity-associated markers, and reproductive hormones and lipids levels in serum and seminal plasma. METHODS: In our prospective study, a total of 1 010 subfertile men, aged from 18 to 50 years old, were enrolled from August 2012 through June 2015. Their obesity-associated markers, semen parameters, sperm acrosomal enzyme activity, seminal plasma biochemical markers, and reproductive hormones and lipids levels in serum and seminal plasma were detected. Sperm DNA fragmentation index (DFI) was determined by sperm chromatin structure assay. The correlations between DFI and each of the above-mentioned variables were analyzed. RESULTS: Spearman correlation analysis showed that sperm DFI was positively related to age and abstinence time (P<0.001). Sperm DFI was also positively related to semen volume and percent of abnormal sperm head (P<0.001), while negatively related to sperm concentration, progressive motility (PR), sperm motility, total normal-progressively motile sperm count (TNPMS), percent of normal sperm morphology (NSM), percent of intact acrosome and acrosomal enzyme activity (P<0.001). Sperm DFI was positively related to seminal plasma zinc level (P<0.001) but unrelated to seminal plasma total α-glucotase, γ-glutamyl transpeptidase (GGT) and fructose levels. There was no any correlation between sperm DFI and obesity-associated markers such as body mass index (BMI), waist-to-hip ratio (WHR), waist circumference (WC) and waist-to-height ratio (WHtR), and serum lipids levels, but there was positive correlation between sperm DFI and seminal plasma triglyceride (TG) and total cholesterol (TC) levels (P<0.001). Sperm DFI was positively related to serum luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels and seminal plasma FSH and estradiol (E2) levels (P<0.001), but unrelated to serum and seminal plasma testosterone (T) levels. The multivariate regression analysis for the variables which were significantly correlated with sperm DFI in Spearman correlation analysis showed that age, semen volume, sperm concentration, progressive motility, TNPMS and intact acrosome were independently correlated with sperm DFI. CONCLUSIONS: There are many potential factors associated with sperm DFI, including age, abstinence time, spermatogenesis and maturation, seminal plasma lipids and reproductive hormones levels. However, the potential effects of seminal plasma lipids and reproductive hormones on sperm DNA damage need still to be demonstrated by the studies with scientific design and a large size of samples.

Advanced glycation end product concentrations in follicular fluid of women undergoing IVF/ICSI with a GnRH agonist protocol.

The accumulation of advanced glycation end products (AGE) is associated with ovarian dysfunction. This study examines whether the accumulation of AGE in follicular fluid affects ovarian responsiveness and embryo quality during IVF/intracytoplasmic sperm injection (IVF/ICSI) with a gonadotrophin-releasing hormone (GnRH) agonist protocol. The levels of AGE in follicular fluid were measured in 127 women undergoing IVF/ICSI in GnRH agonist cycles. Plasma hormones were also measured. Embryos were graded using standard approaches. There were inverse associations between follicular fluid AGE concentration and number of oocytes retrieved, number of fertilized oocytes, number of high-quality embryos, fertilization rate and high-quality embryo rate, adjusted for potential confounders. AGE concentration in follicular fluid was significantly higher in women with an ovarian response below the target (<7 oocytes) compared with those reaching the target (7-15 oocytes) or above the target (>15 oocytes). The cut-off value of 15.3 µg/ml for follicular fluid AGE showed 84.6% sensitivity and 55.5% specificity in evaluating the response to ovarian stimulation as below the target. The results suggest that ovarian responsiveness and embryo quality are related to intraovarian exposure to AGE.

[Value of sperm acrosin activity detection in selecting the method of assisted reproduction for patients with unexplained infertility].

OBJECTIVE: To investigate the clinical significance of sperm acrosin activity detection in selecting the method of assisted reproduction for patients with unexplained infertility (UI). METHODS: This retrospective study included 49 UI couples treated by IVFET (49 cycles) after three failures in intrauterine insemination (IUI) and another 95 couples with uterine tube obstruction (UTO) treated by IVF (131 cycles). We analyzed the laboratory data, clinical outcomes and sperm acrosin activity in the two groups of patients. According to the level of sperm acrosin activity of the males, we further divided the UI patients into two subgroups, a < 36 IU/106 sperm group (20 cycles) and a ≥36 IU/106 sperm group (29 cycles), and compared the fertilization rates between the two groups. RESULTS: Compared with UI couples treated by IVFET, the UTO couples treated by IVF had a significantly lower rate of fertilization (67.0% vs 76.4%, P < 0.05) and a higher rate of remedial intracytoplasmic sperm injection (ICSI) (20.4% vs 6.1%, P < 0.05), but showed no statistically significant differences in the rates of MII oocytes, available embryos, highquality embryos, implantation, and clinical pregnancy from the latter group (P >0.05). The sperm acrosin activity was remarkably lower in the UI than in the UTO patients (36.03 vs 61.98 IU/106, P < 0.01), and so was the fertilization rate in the < 36 IU/106 than in the ≥36 IU/106 sperm subgroup (47.7% vs 80.3%, P < 0.01). CONCLUSIONS: The low fertilization rate caused by decreased sperm acrosin activity may be the main cause of infertility and the potential factor of UI. When sperm acrosin activity is < 36 IU/106 sperm, IVF plus shortterm fertilization by remedial ICSI should be preferred to IUI.