Objective:Analyzing the influence of morphological evaluation parameters of blastocysts, including days of blastocyst development [day 5 (D5) and day 6 (D6)], degree of blastocyst expansion (4, 5, 6), inner cell mass and trophectoderm grade, on the occurrence of chromosomal karyotype abnormalities of chorionic villi in missed abortion after in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment and fresh/frozen-thawed single blastocyst transfer. Methods:The clinical data of patients with missed abortion after IVF/ICSI treatment and fresh/frozen-thawed single blastocyst transfer from February 2015 to February 2023 in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University were included. Using a case-control study, the data were divided into two groups according to the detection results of chromosomal copy number variations (CNVs) in chorionic villi of missed abortion abnormal karyotype group ( n=139) and normal karyotype group ( n=82). The baseline data between the two groups were compared. Univariate logistic regression was used to investigate the effect of blastocyst morphological rating parameters on the occurrence of chromosomal karyotype abnormalities of chorionic villi in aborted tissues, and multivariate logistic regression was also used to adjust confounding factors. Results:Male age [(34.12±6.49) years], sperm morphology rate [5.00 (4.00,6.00)%] and female age [33.00 (30.00, 37.00) years] in abnormal karyotype group were higher than those in the normal karyotype group [(32.38±4.69) years, 4.00 (2.00,5.00)% and 31.50 (29.00,34.00) years], and the differences were statistically significant ( P=0.022, P=0.020, P=0.009). Univariate and multivariate logistic regression analyses showed that days of blastocyst development, degree of blastocyst expansion, inner cell mass and trophectoderm grade did not increase the risk of chromosomal karyotype abnormalities of chorionic villi (all P>0.05). Conclusion:There is no significant correlation between blastocyst morphological evaluation parameters and chromosomal karyotype abnormalities in chorionic villi of missed abortion after fresh/frozen-thawed single blastocyst transfer with IVF/ICSI treatment.

Objective:To explore the key copy number variation (CNV) regions, abortion candidate genes and signaling pathways associated with recurrent spontaneous abortion (RSA).Methods:A retrospective cohort study was conducted based on the data of 1 870 miscarriage cases of RSA patients who received CNV analysis by high-throughput sequencing technology in the Laboratory Medicine Department of the Third Affiliated Hospital of Zhengzhou University from January 2016 to September 2022. These cases were divided into different groups based on the age of miscarriage and gestational age of the pregnant women. Chi-square test or Fisher's exact test was used to analyze the distribution of chromosome abnormalities and CNV. Gene functions and signaling pathways in RSA-related CNV were identified by gene enrichment analysis.Results:Among the 1 870 tissues, 1 001 (53.53%) cases were detected with chromosomal abnormalities. A total of 140 CNVs were detected in 93 tissues (9.29%), including 34 submicroscopic CNVs (segment<10 Mb) and 106 large CNVs with segment≥10 Mb. Submicroscopic pathogenicity CNVs with statistical differences were involved 1p36.33p36.23, 2q37.3, 4p16.3, 22q11.21 (χ 2=6.99, P=0.008) in early RSA embryos (≤12 weeks). 16p11.2 and Xp11.23p11.22 microdeletion were firstly reported in abortion cases. Significantly recurrent large CNVs were mainly involved 18q22q23 (del/dup), 4p16p15, 9p24p22, 8p23p22, and Xp22.3 regions, and the candidate genes mainly concentrated on PI3K-Akt and JAK-STAT signaling pathway. Conclusion:Rare submicroscopic CNVs and recurrent large CNVs were associated with RSA in early pregnancy. GO and KEGG database analysis revealed potential abortion candidate genes and signaling pathways, providing new information for the genetic etiology of RSA.

Objective:Analyzing the influence of morphological evaluation parameters of blastocysts, including days of blastocyst development [day 5 (D5) and day 6 (D6)], degree of blastocyst expansion (4, 5, 6), inner cell mass and trophectoderm grade, on the occurrence of chromosomal karyotype abnormalities of chorionic villi in missed abortion after in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment and fresh/frozen-thawed single blastocyst transfer. Methods:The clinical data of patients with missed abortion after IVF/ICSI treatment and fresh/frozen-thawed single blastocyst transfer from February 2015 to February 2023 in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University were included. Using a case-control study, the data were divided into two groups according to the detection results of chromosomal copy number variations (CNVs) in chorionic villi of missed abortion abnormal karyotype group ( n=139) and normal karyotype group ( n=82). The baseline data between the two groups were compared. Univariate logistic regression was used to investigate the effect of blastocyst morphological rating parameters on the occurrence of chromosomal karyotype abnormalities of chorionic villi in aborted tissues, and multivariate logistic regression was also used to adjust confounding factors. Results:Male age [(34.12±6.49) years], sperm morphology rate [5.00 (4.00,6.00)%] and female age [33.00 (30.00, 37.00) years] in abnormal karyotype group were higher than those in the normal karyotype group [(32.38±4.69) years, 4.00 (2.00,5.00)% and 31.50 (29.00,34.00) years], and the differences were statistically significant ( P=0.022, P=0.020, P=0.009). Univariate and multivariate logistic regression analyses showed that days of blastocyst development, degree of blastocyst expansion, inner cell mass and trophectoderm grade did not increase the risk of chromosomal karyotype abnormalities of chorionic villi (all P>0.05). Conclusion:There is no significant correlation between blastocyst morphological evaluation parameters and chromosomal karyotype abnormalities in chorionic villi of missed abortion after fresh/frozen-thawed single blastocyst transfer with IVF/ICSI treatment.

Objective:To explore the key copy number variation (CNV) regions, abortion candidate genes and signaling pathways associated with recurrent spontaneous abortion (RSA).Methods:A retrospective cohort study was conducted based on the data of 1 870 miscarriage cases of RSA patients who received CNV analysis by high-throughput sequencing technology in the Laboratory Medicine Department of the Third Affiliated Hospital of Zhengzhou University from January 2016 to September 2022. These cases were divided into different groups based on the age of miscarriage and gestational age of the pregnant women. Chi-square test or Fisher's exact test was used to analyze the distribution of chromosome abnormalities and CNV. Gene functions and signaling pathways in RSA-related CNV were identified by gene enrichment analysis.Results:Among the 1 870 tissues, 1 001 (53.53%) cases were detected with chromosomal abnormalities. A total of 140 CNVs were detected in 93 tissues (9.29%), including 34 submicroscopic CNVs (segment<10 Mb) and 106 large CNVs with segment≥10 Mb. Submicroscopic pathogenicity CNVs with statistical differences were involved 1p36.33p36.23, 2q37.3, 4p16.3, 22q11.21 (χ 2=6.99, P=0.008) in early RSA embryos (≤12 weeks). 16p11.2 and Xp11.23p11.22 microdeletion were firstly reported in abortion cases. Significantly recurrent large CNVs were mainly involved 18q22q23 (del/dup), 4p16p15, 9p24p22, 8p23p22, and Xp22.3 regions, and the candidate genes mainly concentrated on PI3K-Akt and JAK-STAT signaling pathway. Conclusion:Rare submicroscopic CNVs and recurrent large CNVs were associated with RSA in early pregnancy. GO and KEGG database analysis revealed potential abortion candidate genes and signaling pathways, providing new information for the genetic etiology of RSA.

Objective:To preliminarily explore the related factors that affect chimera mosaicism in in vitro fertilization (IVF) treatment. Methods:A case-control study was conducted to retrospectively analyze the clinical data of 2252 blastocysts in 579 preimplantation genetic testing (PGT) cycles in the Reproductive Medicine Center of the Third Affiliated Hospital of Zhengzhou University from January 2017 to December 2020. Biopsy cells were analyzed by next generation sequencing (NGS). According to the analysis results, all embryos were divided into mosaicism group and non-mosaicism group. Mosaicism types included euploid-aneuploid mosaicism, aneuploid-aneuploid mosaicism and complex mosaicism. The population characteristics and laboratory-related parameters of the two groups of embryos were compared, and single-factor and multi-factor analysis of the incidence of mosaicism were performed to evaluate the related factors that affect the development of mosaic embryos.Results:A total of 2252 blastocysts in 579 cycles were included in this study, 905 embryos (40.2%) were euploid, 923 (41.0%) were aneuploid, and 424 (18.8%) were mosaicism. Among them, 228 (10.1%) were euploid-aneuploidy mosaicism, 59 (2.6%) were aneuploidy-aneuploidy mosaicism, and 137 (6.1%) were complex mosaicism. NGS technology was performed in 4 institutions, and the mosaicism rate fluctuated between 7.6% and 26.2%. After adjusting the confounding factors (the age of the male and female partners, the quality of the male partner's sperm, the ovarian stimulation protocols, the type of culture medium, the indications of PGT, the different biopsy operators and the developmental stage of the blastocyst), it was found that the blastocyst trophectoderm cell (TE) score (grade C vs. grade A, P=0.014) and the genetic testing institutions (institution 2 vs. early stage of institution 1, P<0.001; late stage of institution 1 vs. early stage of institution 1, P<0.001) had a significant effect on the occurrence of mosaicism. Compared with the TE score of grade A, the chance of mosaicism in grade C increased by 66% (a OR=1.66, 95% CI=1.11-2.50, P=0.014). Compared with the early stage of institution 1, the incidence of mosaicism in institution 2 and late stage of institution 1 was 2.28 times (a OR=2.28, 95% CI=1.71-3.04, P<0.001), and late stage of institution 1 was 2.17 times that of the early stage (a OR=2.17, 95% CI=1.41-3.34, P<0.001). Conclusion:The incidence of mosaicism during IVF treatment is related to NGS genetic testing institutions and the quality of trophectoderm cells

Objective:To preliminarily explore the related factors that affect chimera mosaicism in in vitro fertilization (IVF) treatment. Methods:A case-control study was conducted to retrospectively analyze the clinical data of 2252 blastocysts in 579 preimplantation genetic testing (PGT) cycles in the Reproductive Medicine Center of the Third Affiliated Hospital of Zhengzhou University from January 2017 to December 2020. Biopsy cells were analyzed by next generation sequencing (NGS). According to the analysis results, all embryos were divided into mosaicism group and non-mosaicism group. Mosaicism types included euploid-aneuploid mosaicism, aneuploid-aneuploid mosaicism and complex mosaicism. The population characteristics and laboratory-related parameters of the two groups of embryos were compared, and single-factor and multi-factor analysis of the incidence of mosaicism were performed to evaluate the related factors that affect the development of mosaic embryos.Results:A total of 2252 blastocysts in 579 cycles were included in this study, 905 embryos (40.2%) were euploid, 923 (41.0%) were aneuploid, and 424 (18.8%) were mosaicism. Among them, 228 (10.1%) were euploid-aneuploidy mosaicism, 59 (2.6%) were aneuploidy-aneuploidy mosaicism, and 137 (6.1%) were complex mosaicism. NGS technology was performed in 4 institutions, and the mosaicism rate fluctuated between 7.6% and 26.2%. After adjusting the confounding factors (the age of the male and female partners, the quality of the male partner's sperm, the ovarian stimulation protocols, the type of culture medium, the indications of PGT, the different biopsy operators and the developmental stage of the blastocyst), it was found that the blastocyst trophectoderm cell (TE) score (grade C vs. grade A, P=0.014) and the genetic testing institutions (institution 2 vs. early stage of institution 1, P<0.001; late stage of institution 1 vs. early stage of institution 1, P<0.001) had a significant effect on the occurrence of mosaicism. Compared with the TE score of grade A, the chance of mosaicism in grade C increased by 66% (a OR=1.66, 95% CI=1.11-2.50, P=0.014). Compared with the early stage of institution 1, the incidence of mosaicism in institution 2 and late stage of institution 1 was 2.28 times (a OR=2.28, 95% CI=1.71-3.04, P<0.001), and late stage of institution 1 was 2.17 times that of the early stage (a OR=2.17, 95% CI=1.41-3.34, P<0.001). Conclusion:The incidence of mosaicism during IVF treatment is related to NGS genetic testing institutions and the quality of trophectoderm cells