Objective: To assess the impact of the sperm swim-up method on embryo aneuploidy rates patients undergoing Non-Invasive Chromosomal Screening in conventional in vitro fertilization. Design: Retrospective cohort study. Setting: Hospita district, China Population or Sample: Patients Undergoing Non-Invasive Chromosomal Screening in Conventional In Vitro Fertilization. Methods: A retrospective analysis was conducted on patients who underwent conventional IVF and NICS at our hospital from January 2022 to May 2024. Patients were grouped based on the semen processing method used: the SU group and the DGC group. We compared semen parameters before and after processing on the day of oocyte retrieval, as well as the number of oocytes retrieved, MII oocytes, and two pronuclei embryos. The fertilization rate, cleavage rate, Day 3 high-quality embryo rate, embryo ploidy rate, and aneuploidy rate between the two groups were also analyzed. Main Outcome Measures:embryo ploidy and aneuploidy rate Results: There were no significant differences between the DGC and SU groups in terms of the number of oocytes retrieved, number of MII oocytes, number of normal fertilizations, normal fertilization rate, abnormal fertilization rate, number of cleavages, cleavage rate, blastocyst formation rate, or blastocyst grading (all P > 0.05). However, the DGC group had a significantly higher D3 high-quality embryo rate than the SU group (53.9% vs. 48.20%, P = 0.011). The proportion of embryos rated as Grade A by NICS was significantly lower in the DGC group (31.8%, 262/824) compared to the SU group (39.4%, 119/302) ( P = 0.017). The euploidy rate was 21.7% (179/824) and the aneuploidy rate was 71.2% (587/824) in the DGC group; in the SU group, the euploidy rate was 25.8% (78/302) and the aneuploidy rate was 68.9% (208/302), with no significant differences between the groups ( P > 0.05). Conclusion: In this study, embryos derived from semen processed using the SU method had a higher proportion of Grade A embryos according to NICS compared to those processed by DGC. However, there was no reduction in the embryo aneuploidy rate.

Background: Severe congenital neutropenia (SCN) is an autoimmune disease clinically characterized by persistent and severe neutropenia. It is typically caused by a heterozygous mutation in the ELANE gene, which encodes neutrophil elastase. Procedure: Using one 3-year-old male diagnosed with severe congenital neutropenia, we aimed to assess the clinical and genetic characteristics of SCN caused by ELANE mutations and test whether the mutation can be corrected by CRISPR/Cas9-mediated gene editing. The proband underwent extensive clinical assessments, with exome sequencing and bioinformatics analysis to identify pathogenic genes. In addition, Sanger sequencing was used to verify the pedigrees. The cell line, 293-ELANE, harboring ELANE mutation was generated by lentiviral transduction and selection with puromycin from HEK-293. And the mutation was corrected by CRISPR/Cas9-mediated homology-directed repair (HDR). Results: The ELANE gene test in the proband unveiled a heterozygous de novo missense mutation: c. 248T>A (p.V83D), which is not detected in his asymptomatic parents with peripheral blood samples. Surprisingly, we found that 46.01% of his father’s sperm cells had the same mutation. These results demonstrated that the proband inherited the ELANE mutation from his father, who had a normal neutrophil count but was germline mosaic. In this study, the highest repair efficiency of CRISPR/Cas9-mediated HDR for 293-ELANE is 4.43%. Conclusions: We identified a missense mutation (p.V83D) in ELANE causing SCN, the first report on paternal semen mosaicism of an ELANE mutation. Our study also paves the way for preimplantation genetic diagnosis (PGD) based on ELANE mutation prevention and clinical treatment of congenital disabilities.