OBJECTIVE: To explore the genetic etiology and related factors in 1 065 women with spontaneous abortions. METHODS: All patients have presented at the Center of Prenatal Diagnosis of Nanjing Drum Tower Hospital from January 2018 to December 2021. Chorionic villi and fetal skin samples were collected, and the genomic DNA was assayed by chromosomal microarray analysis (CMA). For 10 couples with recurrent spontaneous abortions but normal CMA results for abortive tissues, non-in vitro fertilization-embryo transfer (IVF-ET) pregnancies and no previous history of live births and no structural abnormalities of the uterus, peripheral venous blood samples were collected. Genomic DNA was subjected to trio-whole exome sequencing (trio-WES). Candidate variants were verified by Sanger sequencing and bioinformatics analysis. Multifactorial unconditional logistic regression analysis was carried out to analyze the factors that may affect chromosomal abnormality in spontaneous abortions, such as the age of the couple, number of previous spontaneous abortions, IVF-ET pregnancy and history of live birth. The incidence of chromosomal aneuploidies in spontaneous abortions during the first trimester was compared in young or advanced-aged patients by chi-square test for liner trend. RESULTS: Among the 1 065 spontaneous abortion patients, 570 cases (53.5%) of chromosomal abnormalities were detected in spontaneous abortion tissues, which included 489 cases (45.9%) of chromosomal aneuploidies and 36 cases (3.4%) of pathogenic/likely pathogenic copy number variations (CNVs). Trio-WES results have revealed one homozygote variant and one compound heterozygote variants in two pedigrees, both of which were inherited from the parents. One likely pathogenic variant was detected in the patient from two pedigrees. Multifactorial unconditional Logistic regression analysis suggested that age of patient was an independent risk factor of chromosome abnormalities (OR = 1.122, 95%CI: 1.069-1.177, P < 0.001), the number of previous abortions and IVF-ET pregnancy were independent protective factors for chromosomal abnormalities (OR = 0.791, 0.648; 95%CI: 0.682-0.916, 0.500-0.840; P = 0.002, 0.001), whilst the age of husband and history of live birth were not (P > 0.05). The incidence of aneuploidies in the abortive tissues has decreased with the number of previous spontaneous abortions in young patients (χ² = 18.051, P < 0.001), but was not significantly correlated with the number of previous spontaneous abortions in advanced-aged patients with spontaneous abortions (P > 0.05). CONCLUSION Chromosomal aneuploidy is the main genetic factor for spontaneous abortion, though CNVs and genetic variants may also underlie its genetic etiology. The age of patients, number of previous abortions and IVF-ET pregnancy are closely associated with chromosome abnormalities in abortive tissues.
Pregnancy ; Humans ; Female ; Aged ; Abortion, Spontaneous/genetics* ; DNA Copy Number Variations ; Chromosome Aberrations ; Chromosome Disorders/genetics* ; Aneuploidy ; Abortion, Habitual/genetics*

OBJECTIVES: The mechanism for traditional Chinese medicine in treating of recurrent spontaneous abortion is not clear. This study aims to explore the mechanism of baotaiyin in the treatment of recurrent abortion by regulating the immune inflammatory axis of interleukin (IL)-23/helper T cell (Th)17. METHODS: Spontaneous abortion model mice were randomly divided into a model group, 3 dose (low, medium, and high) groups of baotaiyin, with 10 mice in each group. After 14 days of medication, the levels of IL-17, IL-23, IL-10, and TGF-β in serum were detected with enzyme-linked immunosorbent assay. The proportion of Th17 and regulatory T cells (Treg) cells in spleen lymphocytes was tested with flow cytometry. The expressions of (retinoid-related orphan receptor γt, ROR-γt) and forkhead box P3 (FOXP3) mRNA in decidua tissues was detected with RT-PCR. Embryo absorption rate was counted. RESULTS: Compared with the model group, the absorption rate of embryo and Th17/Treg cell ratio in baotaiyin medium- and high-dose groups were decreased significantly (all P<0.05); the levels of IL-17 and IL-23 in serum were decreased (both P<0.05), while the levels of TGF-β and IL-10 in baotaiyin medium- and high-dose groups were increased (P<0.05, P<0.01, respectively); the expression of ROR-γt mRNA was decreased and the expression of FOXP3 mRNA was increased (all P<0.01) in decidua tissues of baotaiyin medium- and high-dose groups. CONCLUSIONS Baotaiyin inhibits the positive feedback cycle of IL-23/Th17 immune inflammatory axis, which regulates Th17/Treg cell balance, mediates the maternal and fetal immune tolerance, and prevents the recurrent abortion.
Mice ; Animals ; Female ; Humans ; Pregnancy ; Interleukin-23 ; Interleukin-17/genetics* ; Interleukin-10 ; Abortion, Habitual ; Transforming Growth Factor beta/genetics*

Here we describe a Syrian couple having recurrent pregnancy loss in the first trimester, fetal malformations, and/or neonatal death. The father had a balanced chromosomal translocation t(5;15), an sY125 microdeletion of locus b in the azoospermia factor (AZF) gene, and an MTHFR C677T homozygous polymorphism with normal phenotype. Interestingly, his healthy wife had another MTHFR A1298C homozygous polymorphism. The couple experienced two pregnancy losses and had two stillborn children with severe malformations due to partial trisomy of the short arm of chromosome 5. The couple does not have any living offspring after 10 years of marriage.
Abortion, Habitual ; genetics ; Azoospermia ; genetics ; Chromosome Aberrations ; Chromosomes, Human, Pair 5 ; Female ; Fetal Death ; etiology ; Homozygote ; Humans ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Polymorphism, Genetic ; Pregnancy ; Translocation, Genetic ; Trisomy

BACKGROUNDGenetic factors are the main cause of early miscarriage. This study aimed to investigate aneuploidy in spontaneous abortion by fluorescence in situ hybridization (FISH) using probes for 13, 16, 18, 21, 22, X and Y chromosomes.

METHODSA total of 840 chorionic samples from spontaneous abortion were collected and examined by FISH. We analyzed the incidence and type of abnormal cases and sex ratio in the samples. We also analyzed the relationship between the rate of aneuploidy and parental age, the rate of aneuploidy between recurrent abortion and sporadic abortion, the difference in incidence of aneuploidy between samples from previous artificial abortion and those from no previous induced abortion.

RESULTSA total of 832 samples were finally analyzed. 368 (44.23%) were abnormal, in which 84.24% (310/368) were aneuploidies and 15.76% (58/368) were polyploidies. The first was trisomy 16 (121/310), followed by trisomy 22, and X monosomy. There was no significant difference in the rate of aneuploidy in the advanced maternal age group (≥ 35 years old) and young maternal age group (<35 years old). However, the rate of trisomy 22 and the total rate of trisomies 21, 13, and 18 (the number of trisomy 21 plus trisomy 13 and trisomy 18 together) showed significantly different in two groups. We found no skewed sex ratio. There was no significant difference in the rate of aneuploidy between recurrent miscarriage and sporadic abortion or between the samples from previous artificial abortion and those from no previous artificial abortion.

CONCLUSIONSAneuploidy is a principal factor of miscarriage and total parental age is a risk factor. There is no skewed sex ratio in spontaneous abortion. There is also no difference in the rate of aneuploidy between recurrent abortion and sporadic abortion or between previous artificial abortion and no previous induced abortion.

Abortion, Habitual ; genetics ; Abortion, Spontaneous ; genetics ; Adult ; Aneuploidy ; Female ; Humans ; In Situ Hybridization ; Middle Aged ; Pregnancy ; Sex Ratio

OBJECTIVETo analyze the characteristics of complex chromosomal rearrangement (CCR) in Chinese male carriers and its influence on male fertility.

METHODSUsing the G band technique, we conducted karyotype analysis on the peripheral blood lymphocytes of 1,625 Chinese males with reproductive problems. We also searched CNKI and Wanfang database for CCR-related literature published between January 1984 and November 2013, followed by statistical analysis on the CCR characteristics and reproduction-related data of the CCR carriers.

RESULTSTwo CCR carriers were found among the 1,625 males and another 47 cases identified from the databases. Among the 49 CCR carriers, there were 17 three-way exchange cases (34.7%), 17 double two-way exchange cases (34.7%), and 15 exceptional cases (30.6%), with no statistically significant differences in the incidence of the three types (P > 0.05). Azoospermia- or oligospermia-induced infertility was found in 19 (38.8% ) of the CCR carriers. A total of 87 pregnancies were achieved in the other 30 (61.2%), among which spontaneous abortion occurred in 75.9% (66/87), dead fetus and malformed infant death in 9.2% (8/87), and phenotypically normal offspring in 14.9% (13/87). Recurrent abortion was associated frequently with breakpoints on CCR-involved chromosomes 6, 7, 8, 11, and 16, while dyszoospermia mostly with breakpoints on CCR-involved chromosomes 10 and 14. The breaking occurred more than 3 times at 1p22, 1q25, 2q31, 5p13, 5q35, 6q23, 8q13, and 20p13. Moreo- ver, the breakpoints at 2q31, 5q35, and 8q13 were particularly related to recurrent abortion, while that at 1p22 only to dyszoospermia.

CONCLUSIONCCR is extremely rare. Male CCR carriers are often identified through reproductive problems and have high risks of infertility and abnormal pregnancy and a very low rate of normal newborns. In addition, chromosomes and breakpoints involved in CCR may affect the fertility of male CCR carriers, and some particular chromosomal breakpoints may play a key role in gametogenesis.

Abortion, Habitual ; Azoospermia ; genetics ; Chromosome Aberrations ; Chromosome Banding ; Chromosome Breakpoints ; Female ; Fertility ; genetics ; Heterozygote ; Humans ; Infertility, Male ; genetics ; Karyotyping ; Male ; Oligospermia ; genetics ; Pregnancy ; Reproduction ; Translocation, Genetic

OBJECTIVETo investigate the association between IL-18 polymorphisms and the risk of unexplained recurrent spontaneous abortion (URSA).

METHODSThe polymorphism of rs187238, rs360718 and rs360717 in IL-18 was determined by PCR in combination with DNA sequencing in 207 patients with URSA and 144 women with normal pregnancy.

RESULTSThe frequency of gene types GG, GC+CC of rs187238 (-137 G/C) in URSA group and control group was 77.3%, 22.7%, and 95.8%, 4.2%, respectively (χ²=22.767, P<0.001). The frequency of allele C in URSA group was significantly higher than that in control groups (13.04% vs 2.1%, χ²=26.102, P<0.001) . The risk of spontaneous abortion in C allele carriers was 7.050 times higher than that in G allele carriers (OR=7.050, 95%CI: 2.990-16.622). No significant difference in genotype frequency and allele frequency of rs360718 and rs360717 polymorphism was noticed between URSA group and control group (χ²=1.497, P=0.221; χ²=0.858, P=0.354).

CONCLUSIONGC+CC genotype and C allele of Rs187238 in IL-18 gene are associated with the susceptibility of recurrent spontaneous miscarriage. Rs360718 and rs360717 in IL-18 may not be associated with URSA.

Abortion, Habitual ; genetics ; Adult ; Female ; Genetic Predisposition to Disease ; Humans ; Interleukin-18 ; genetics ; Polymorphism, Genetic ; Young Adult

OBJECTIVETo evaluate and compare standard sperm parameters and sperm DNA fragmentation in seminal ejaculates from men whose partners had a history of recurrent pregnancy loss (RPL) and a control group of men who had recently established their fertility.

METHODSSemen samples from 85 patients with a history of RPL and 20 men with proven fertility were analyzed according to World Health Organization guidelines. Sperm DNA fragmentation was detected by sperm chromatin dispersion test (SCD).

RESULTSA significant difference (P< 0.05) was observed in sperm motility but not other parameters between the two groups. The mean number of sperm cells with fragmented DNA, represented as DNA fragmentation index, was significantly increased in the RPL group [(34.99± 14.62)%] compared with controls [(10.82± 4.80)%].

CONCLUSIONThis study has indicated that sperm from men with a history of RPL have a higher incidence of DNA damage and poor motility compared with fertile males.

Abortion, Habitual ; etiology ; genetics ; Adult ; DNA Damage ; DNA Fragmentation ; Female ; Humans ; Male ; Pregnancy ; Sperm Motility

BACKGROUNDExperimental evidence indicates that cyclooxygenase-2 (COX-2) plays a critical role in blastocyst implantation; however, little is known of the role of COX-2 in unexplained recurrent spontaneous abortion (URSA).

METHODSWe evaluated the expression level and potential signaling pathway of COX-2 in 30 cases of URSA who were excluded the abnormality of chromosomes, anatomy, endocrine, infectious, autoimmune diseases and in 30 normal pregnancies.

RESULTSThe mRNA and the protein expression level of COX-2 in the URSA group (-0.238 +/- 0.848, 0.368 +/- 0.089, respectively) were significantly lower than that in the control group (1.943 +/- 3.845, 1.046 +/- 0.108, respectively) (both, P < 0.01). The expression of prostaglandins PGF(2a), PGD(2), PGE(2), and PGI(2), in the URSA group ((2326.0 +/- 295.6) pg/ml, (2164.0 +/- 240.5) pg/ml, (238.7 +/- 26.4) pg/ml, (2337.0 +/- 263.0) pg/ml, respectively) were significantly lower than that in the control group ((3450.0 +/- 421.7) pg/ml, (3174.0 +/- 415.6) pg/ml, (323.5 +/- 43.8) pg/ml, (3623.0 +/- 460.4) pg/ml, respectively) (P < 0.05). The mRNA expression level of PPARbeta and RXRalpha (0.859 +/- 0.653, -0.172 +/- 0.752, respectively) in URSA group was significantly lower than that in the control group (1.554 +/- 1.735, 0.777 +/- 2.482, respectively) (both P< 0.05). The mRNA and protein expression levels of vascular endothelial growth factor-A (VEGF-A) in the URSA group (2.010 +/- 1.522, 0.35 +/- 0.46) was significantly lower than that in the control group (4.569 +/- 2.430, 0.750 +/- 0.350) (both P < 0.05).

CONCLUSIONSCOX-2 and the COX-2-derived PGI(2) signaling pathway possibly play an important role in successful embryo implantation, and their decreased expression may result in URSA. The decreased expression may influence the expression of VEGF-A which interferes with placental angiogenesis causing failure of embryo implantation, leading to spontaneous abortion.

Abortion, Habitual ; enzymology ; genetics ; Adult ; Blotting, Western ; Cyclooxygenase 2 ; genetics ; metabolism ; Dinoprost ; metabolism ; Dinoprostone ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Epoprostenol ; metabolism ; Female ; Humans ; Polymerase Chain Reaction ; Pregnancy ; Prostaglandin D2 ; metabolism ; Signal Transduction ; genetics ; physiology ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo study the relationship between azoospermia factor (AZF) microdeletions of the Y-chromosome and recurrent spontaneous abortion.

METHODSWe collected 26 chorionic villus samples from abortive male embryos and 51 blood samples from the husbands whose wives had recurrent spontaneous abortion, extracted genomic DNA from the samples and detected 12 STSs in the AZFa, AZFb and AZFc regions of Yq11.2 by amplification multiplex PCR.

RESULTAZF microdeletions were found neither in the chorionic villus samples nor in the blood samples.

CONCLUSIONThere is no relationship between AZF microdeletions and recurrent spontaneous abortion.

Abortion, Habitual ; genetics ; Abortion, Spontaneous ; genetics ; Chromosome Deletion ; Chromosomes, Human, Y ; Female ; Genetic Loci ; Humans ; Male ; Polymerase Chain Reaction ; methods ; Pregnancy ; Seminal Plasma Proteins ; genetics

OBJECTIVETo investigate changes in sperm chromosome and sperm DNA integrity of infertile males.

METHODSThe level of DNA fragmentation was determined by Sperm Chromatin Dispersion (SCD) test in infertile males with idiopathic severe oligoasthenozoospermia (ISOA, n= 19), couples with unexplained recurrent miscarriage (URM, n= 38) and adult healthy fertile men (control group, n= 32). Multi-color fluorescence in situ hybridization (FISH) was performed with probes specific for chromosomes 13, 18, 21, X and Y in the control group (n= 5), the ISOA (n= 10) and the URM (n= 12).

RESULTSPatients with ISOA and URM showed a significantly higher abnormality with total rate of 4.02% (n= 19) and 3.91%(n= 38) for chromosomes 13, 18 and 21, and 2.03%, 1.98% for chromosomes X and Y, respectively, in their spermatozoa compared to control (1.29% and 0.61%, P< 0.01). A significantly higher proportion of total sperm DNA fragmentation was detected in patients with ISOA (40.7%+/- 17.8%) and URM (22.1%+/- 10.3%) of sperm compared to the control group (12.1%+/- 5.2%, P< 0.01). Moreover, a positive correlation was found between the rate of sperm chromosomal aberration and the rate of sperm DNA fragmentation (gamma = 0.874, P< 0.01, n= 27). There were significant correlation between sperm DNA fragmentation and sperm density, sperm motility and abnormal sperm (gamma = - 0.571, gamma = - 0.616 and gamma = 0.637, respectively, P< 0.01).

CONCLUSIONThe result indicates that spermatozoa from patients with ISOA and URM contain greater DNA fragmentation and chromosomal aneuploidy and may lead to male infertility. Screening for sperm DNA damage may provide useful information in the diagnosis of male idiopathic infertility.

Abortion, Habitual ; pathology ; Adult ; Chromatin ; metabolism ; Chromosome Aberrations ; DNA Fragmentation ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Infertility, Male ; genetics ; pathology ; physiopathology ; Male ; Sperm Count ; Sperm Motility ; Spermatozoa ; metabolism ; pathology