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*

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 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

Karyotypes were prepared from peripheral blood leukocytes in 18 couples with his-tories of habitual abortions. The Standard chromosome analysis and G-banding techniques were studied. The abnormal karyotypes seen were one case with 20% of 45, XX, -14, -15, t(14/15), one case of 46, XY/45, XY, -21 mosaicism, one case of 45, XX, -14, -21, t(14/21), one case of 46, XX/45, XO mosaicism and one case of 46, XYq+. Many other types of chromosomal abnormalities from many reports in couples with spontaneous abortions are discussed.
Abortion, Habitual/genetics* ; Adult ; Chromosome Aberrations* ; Female ; Human ; Leukocytes/ultrastructure ; Male ; Mosaicism ; Pregnancy ; Translocation (Genetics)

We report on a de novo centric fission of chromosome 11 in a healthy female referred for chromosome analysis due to recurrent miscarriages. Both fission products were mitotically stable. This centric fission of chromosome 11 appears to have no clinical significance for this patient other than recurrent miscarriages.
Humans ; Female ; *Chromosomes, Human, Pair 11 ; *Chromosome Aberrations ; Adult ; Abortion, Habitual/*genetics

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

Recurrent spontaneous abortion (RSA) defines as two or more consecutive losses at < or =20 weeks of gestation and affects an estimated 1 of every 100 couples wishing to have children. However, it remains a poorly understood phenomenon. Recent reports observed a significant association between highly skewed X chromosome and RSA, supporting that X chromosome inactivation might be an important and previously unknown cause of RSA. X-inactivation pattern, using polymeric X-linked women with idiopathic RSA and 80 control subjects with a single successful pregnancy and no history of spontaneous abortion. The ratio of heterozygotes was 68.2% (45/66) in women with RSA and 67.5% (54/80) in control group. Among 45 informative RSA cases, only 1 (2.2%) woman showed extreme skewed X inactivation (> or =90%) and 4 (8.9%) had mild skewed inactivation (> or =85%). In 54 heterozygous control subjects, 5 (9.3%) women showed extreme skewed X inactivation and 7 (13.0%) had mild one. The frequency of skewed X inactivation between RSA patients and control group was not significantly different (p>0.05). This finding suggests that skewed x romosome be not associated with unexplained RSA patients.
Abortion, Habitual/*genetics ; Abortion, Spontaneous/*genetics ; Adult ; DNA Methylation ; *Dosage Compensation (Genetics) ; Female ; Heterozygote ; Human ; Korea ; Linkage (Genetics) ; Lymphocytes ; Pregnancy ; Support, Non-U.S. Gov't

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

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*

OBJECTIVETo explore the association of nitric oxide synthase 3 (NOS3) gene variable number of tandem repeat(VNTR) polymorphism in intron 4 and an 894(G/T) mutation at exon 7 with recurrent early spontaneous abortion (RESA).

METHODSOne hundred and forty RESA women (patient group) and 140 healthy women with at least 1 pregnancy and without a history of pregnancy complications (control group) were included. The genotypes of NOS3 gene VNTR polymorphism were determined by polymerase chain reaction and agarose gel electrophoresis. The 894(G/T) mutation of genotypes of NOS3 gene at exon 7 was assessed by polymerase chain reaction-restrictive fragment length polymorphism.

RESULTSThe frequencies of aa and ba genotypes and a allele of NOS3 gene were higher in patient group than in control group (chi square: 4.51, P< 0.05; chi square: 4.29, P<0.05). The aa and ba genotypes were significantly associated with RESA (OR:1.8, 95% CI: 1.04-3.24). There was no significant difference in TT and GT genotypes and T allele of NOS3 gene between RESA patient group and control group (chi square: 1.16, P> 0.05; chi square:1.12, P> 0.05). 894(G/T) polymorphism may be not associated with RESA.

CONCLUSIONThe genetic polymorphism of NOS3 gene 27 bp VNTR was associated with RESA. The genetic polymorphism of NOS3 gene 894(G/T) may be not associated with RESA. These results support that a allele of the NOS3 gene may be susceptibility allele.

Abortion, Habitual ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Minisatellite Repeats ; genetics ; Nitric Oxide Synthase Type III ; genetics ; Polymorphism, Genetic ; genetics ; Pregnancy ; Time Factors