Recurrent Pregnancy Loss Group Research Articles

Immune Checkpoints in Recurrent Pregnancy Loss: New Insights into a Detrimental and Elusive Disorder.

Recurrent pregnancy loss (RPL) refers to two or more miscarriages before 20 weeks gestation. Its prevalence is 1-2%; its pathogenesis remains unexplained in more than 50% of cases, in which the cause is thought to be abnormal immune activity during placentation leading to a lack of pregnancy-induced immune tolerance. It is unknown whether immune activity is deranged in the endometrium of women with RPL. We studied the gene expression and the quantitative tissue protein levels of three immune checkpoints (CD276, which enhances cytotoxic T-cell activity, cytotoxic T-lymphocyte-associated antigen-4 [CTL-4], which reduces Th1 cytokine production, and lymphocyte activation gene-3 [LAG-3], which shows suppressive activity on Tregs and CD4+ T-cells) in endometrial samples from 27 women with unexplained RPL and in 29 women with dysfunctional uterine bleeding and previous uneventful pregnancies as controls. RNA isolation, real-time PCR, protein isolation, and ELISA were performed. CD276 gene expression and protein tissue levels were significantly lower in the endometrium of the RPL group than in the controls, whereas both CTL-4 and LAG-3 were significantly higher. This difference suggests defective endometrial immune regulation and overactivation of immune response in women with a history of RPL, at least in relation to controls with dysfunctional uterine bleeding and previous normal reproductive history.

Gene Expression Levels of CSF-1 and CSF-1R Endometrial under The Influence of Prolactin Level in Unexplained Miscarriage: A Case-Control Study.

Hormones such as prolactin, by influencing expression of the endometrial genes, play a pivotal role in embryo implantation and development. The present study aimed to evaluate serum level of prolactin and its effect on altering expression level of colony-stimulating factor-1 (CSF-1) and colony-stimulating factor-1 receptor (CSF-1R) genes in endometrial tissue during in vitro fertilization (IVF) pregnancy in the infertile women and recurrent pregnancy loss (RPL), compared to fertile women, who lost their pregnancies at gestational age <20 weeks. In this case-control study, 40 infertile women, 40 IVF pregnant women with RPL and 40 fertile women who lost their pregnancies at <20 weeks of gestation for unknown reasons were selected. Prolactin serum level was assessed using ELISA technique and expression of CSF-1 and CSF-1R genes was determined in endometrial tissue, using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Mean prolactin level of the infertile group was 24.38 ± 1.43 ng/mL and it had statistically significant relationship with the fertile group (P<0.001). Expression level of the CSF-1 and CSF-1R genes were higher in the fertile than infertile groups by 2.88 times (P<0.0001) and 2.64 times (P<0.0001), while it was respectively 2.28 (P<0.0001) and 1.69 (P<0.0001) times higher compared to the RPL group. Risk factors for pregnancy loss, such as aging, increased body mass index (BMI), smoking and diabetes caused decreasing changes in gene expression (CSF-1 and CSF-1R ) and the differences were statistically significant, except in the infertile group. The present study showed a significant relationship of CSF-1 and CSF-1R expression levels with pregnancy loss. Risk factors such as aging, obesity, smoking and diabetes decreased both genes expression levels.

The effect of lymphocyte immunotherapy (LIT) in modulating immune responses in patients with recurrent pregnancy loss (RPL).

In order to prevent miscarriage in RPL patients, the goal of this study was to determine how well lymphocyte immunotherapy (LIT) works in modifying immunological responses produced by cells, cytokines, transcription factors, and microRNAs. 200 RPL patients and 200 healthy controls were included in the study. Using flow cytometry, it was possible to compare the frequency of cells before and after lymphocyte treatment. Real-time PCR was used to assess the gene expression levels of transcription factors, cytokines, and microRNAs. ELISA method was used to evaluate the level of secretion of cytokines in the serum. Primary evaluation of the immune profile between healthy controls and RPL cases showed a higher frequency of Th17, NK, B cells and a lower frequency of Treg cells in RPL cases. Also, pro-inflammatory cytokines showed increased expression at mRNA and protein levels in the RPL group in comparison with the control group. Whereas, anti-inflammatory cytokines showed decreased expression in RPL patients. Decreased and increased frequency of Th17 and Treg lymphocytes observed in RPL cases following LIT, respectively. The same results obtained for RORγt and FoxP3 mRNA expression as transcription factor of Th17 and Treg cells, respectively. NK cell cytotoxicity decreased after LIT in RPL patients. miR-326a and miR-155 expression after LIT reduced, but miR-146a and miR-10a expression increased in RPL instances. LIT in RPL cases causes to elevation and modulation of anti-inflammatory and pro-inflammatory cytokines. Our data showed that lymphocyte therapy can be proposed as an effective therapeutic agent in RPL patients with immunological background by a modulating inflammatory condition.

Association of plasma folic acid, vitamin-B12 and homocysteine with recurrent pregnancy loss. "A case control study".

To investigate the relationship of serum homocysteine, folic acid, and vitamin-B12 levels with recurrent pregnancy loss (RPL). A case-control study was conducted in the Department of Gynecology and Obstetrics, Dr. Ruth Pfau Civil Hospital, Karachi from July 2021 to June 2022. Total 124 participants were recruited from gynecology outpatient department after taking informed consent. The participants included 62 non-pregnant females with two or more consecutive unexplained RPLs and 62 healthy women having at least two successful deliveries without any pregnancy loss. Serum folic acid and vitamin-B12 levels were measured by chemiluminescent method and serum homocysteine was measured by enzyme-linked immunosorbent assay (ELISA). Comparison of quantitative variables with RPL cases and control was done by Mann-Whitney U-test (for non-normally distributed data) and independent sample t-test (for normally distributed data), while Pearson's chi-square test was used for the association of qualitative variables with RPL cases and control. Correlation of homocysteine with vitamin-B12 and folic acid was assessed in RPL cases. The median age of the study population was 27 (IQR 25-32) years. The median body mass index (BMI) was 26.25 (IQR 22-29) kg/m2. Cases had significantly lower serum folic acid (p-value=0.022), while vitamin-B12 was decreased in cases as compared to controls (p-value=0.295). Mean concentration of serum homocysteine was higher in RPL cases as compared to controls (p-value=0.094). There was significant association of serum vitamin B12 (p-value=0.001) and folic acid levels (p-value=0.004) with RPL. The homocysteine was not significantly correlated with vitamin-B12 (r=0.124, p=0.338) and folic acid (r=0.067, p=0.606) in the RPL group. Reduced level of serum folic acid and vitamin B12 was significantly associated with RPL cases, while homocysteine was marginally raised however the difference was not significant. Folic acid and vitamin B12 supplementation before next pregnancy in RPL patients is likely to be beneficial in improving pregnancy outcomes.

Identification of novel biomarkers and immune infiltration features of recurrent pregnancy loss by machine learning

Recurrent pregnancy loss (RPL) is a complex reproductive disorder. The incompletely understood pathophysiology of RPL makes early detection and exact treatment difficult. The purpose of this work was to discover optimally characterized genes (OFGs) of RPL and to investigate immune cell infiltration in RPL. It will aid in better understanding the etiology of RPL and in the early detection of RPL. The RPL-related datasets were obtained from the Gene Expression Omnibus (GEO), namely GSE165004 and GSE26787. We performed functional enrichment analysis on the screened differentially expressed genes (DEGs). Three machine learning techniques are used to generate the OFGs. A CIBERSORT analysis was conducted to examine the immune infiltration in RPL patients compared with normal controls and to investigate the correlation between OFGs and immune cells. Between the RPL and control groups, 42 DEGs were discovered. These DEGs were found to be involved in cell signal transduction, cytokine receptor interactions, and immunological response, according to the functional enrichment analysis. By integrating OFGs from the LASSO, SVM-REF, and RF algorithms (AUC > 0.880), we screened for three down-regulated genes: ZNF90, TPT1P8, FGF2, and an up-regulated FAM166B. Immune infiltration study revealed that RPL samples had more monocytes (P < 0.001) and fewer T cells (P = 0.005) than controls, which may contribute to RPL pathogenesis. Additionally, all OFGs linked with various invading immune cells to varying degrees. In conclusion, ZNF90, TPT1P8, FGF2, and FAM166B are potential RPL biomarkers, offering new avenues for research into the molecular mechanisms of RPL immune modulation and early detection.

P-507 Oxidative/Nitrative Stress Biomarkers Increased the Risk of Recurrent Pregnancy Loss

Abstract Study question Whether oxidative/nitrative stress biomarkers increased the risk of recurrent pregnancy loss (RPL)? Summary answer Patients with higher oxidative/nitrative stress biomarkers of 8-NO2Gua and HNE-MA had an increased risk of recurrent pregnancy loss compared with controls. What is known already In reproductive medicine, recurrent pregnancy loss (RPL) is a major problem including the most unknown and lack of evidence-based diagnostics and therapies. Certain oxidative stress markers play an important role in the progress of some diseases. In one study, subsequent increases in 8-OHdG levels in the RPL group indicated extensive DNA damage and the lipid peroxidation products dramatically increased before an abortion occurred. Large unknowns of other indicators of oxidative stress, such as markers of lipid peroxidation (8-isoPF 2α, HNE-MA, and MDA), and DNA damage (8-NO2Gua), are studied for their impacts on RPL. Study design, size, duration We used an established case-control study, the Taiwan Recurrent Pregnancy Loss and Environmental Study (TRIPLES), which included 514 reproductive age (20-50) women (including 397 cases and 117 controls) from obstetric clinics at National Cheng Kung University Hospital in southern Taiwan from 2013 to 2022. Participants/materials, setting, methods The physicians diagnosed two or more consecutive miscarriages before 20 weeks of pregnancy as RPL cases. They excluded uterine anomalies, maternal diseases, chromosomal abnormalities, polycystic ovarian syndrome (PCOS), premature ovarian failure, and endometriosis. As a control group, we included married women, who delivered at least one child without artificial reproduction, underwent laparotomy for other clinical reasons, and did not suffer from the above-mentioned gynecological diseases. Main results and the role of chance Recurrent pregnancy loss (RPL) groups were older (mean=35.13) and had more time to prepare for pregnancy (mean=10.66 months). Their marital status mainly was married, their household income exceeded around 33,000 USD, and they consumed coffee habits. The median oxidative/nitrative stress biomarkers of 8-NO2Gua and HNE-MA in the case group were significantly higher than those in the control group (6.15 and 30.12 vs. 3.77 and 21.54 ug/L, p&amp;lt;0.001), indicating that the oxidative stress biomarkers of 8-NO2Gua and HNE-MA could be an essential effect factor in RPL. By categorizing the data by tertile of oxidative/nitrative stress biomarkers, we found that the RPL risk was 3.19 times higher in the third tertile of log 8-NO2Gua than in the first tertile (OR: 3.19, 95% CI: 1.66–6.10) and that the RPL risk in log HNE-MA was higher in the third tertile (OR: 2.05, 95% CI: 1.12–3.74). After adjustment for age, time to pregnancy (months), and coffee drinking habits, the RPL risk in the third tertile of log 8-NO2Gua was 2.01 times significantly higher than that in the first tertile (AOR: 2.01, 95% CI: 1.00–4.04). Limitations, reasons for caution Limitation of a case-control study. The oxidative stress biomarkers 8-NO2Gua and HNE-MA were significantly higher in the RPL groups, however, the causes of oxidative stress (like multiple environmental exposures, etc.) remain more studies to elucidate. Wider implications of the findings We provided evidence for oxidative/nitrative stress biomarkers in RPL patients, which implies the reduction of oxidative/nitrative stress may improve the progress of RPL. As important information for preventive medicine, more studies are needed to understand the adverse outcome pathway of oxidative/nitrative stress in RPL. Trial registration number MOST 109-2314-B-400 −022 -MY3 and EM-112-PP-11

Evaluation of Selenium and Vitamin E Levels in Women with Recurrent Miscarriage

The most frequent pregnancy issue is a miscarriage, which is defined as the spontaneous end of pregnancy before the fetus reaches viability. The current research aims to identify the potential significance of selenium (Se) and vitamin E (VE) as predictive diagnostic biomarkers in recurrent pregnancy loss (RPL). Methods: A case control study that included three study groups; 35 women with RPL, 30 non-aborted pregnant (NAP) women, and 25 healthy women. After rigorously applying the eligibility requirements, patients were chosen from those visiting Bint Al-Huda Teaching Hospital and Suq Al-Shuyukh General Hospital in Thi-Qar Province (Iraq). Serum VE and serum Se levels were determined using the ELISA and atomic absorption spectrophotometer techniques, respectively. Results: The outcomes showed that the RPL group's serum VE and Se below normal levels were (88.6% and 62.9% respectively) lower than in the healthy control group (28% and 24% respectively). The level of VE was depleted in NAP group (93.3%) compared to HC group (28%) with significant differences, whereas the level of Se was significantly lower in RPL group (62.9%) compared NAP group (13.3%). Conclusions: A significant positive association of VE and Se in RPL and a significant negative association of VE and Se in NAP and HC.

Prevalence of hysteroscopic alterations in patients with recurrent pregnancy loss

ObjectiveTo determine the prevalence of uterine malformations and endometrial disorders diagnosed through office hysteroscopy in women with recurrent pregnancy loss (RPL). MethodsRetrospective analysis of medical records of 114 patients referred to a tertiary academic hospital in Belo Horizonte, Brazil, between January 2014 and August 2022, due to a history of two or more miscarriages (RPL group) and 1144 women preparing for in vitro fertilization due to couple infertility (control group). All the participants underwent a diagnostic office hysteroscopy in the proliferative phase of the menstrual cycle. The prevalence of hysteroscopic alterations was compared between the RPL and control groups and between women with two versus three or more miscarriages, using the chi-square test. ResultsIn comparison with the control group, the RPL group had a higher prevalence of uterine malformations (15.8% vs. 4.6%, p<0.001), endometrial adhesions (12.3% vs. 3.3%, p<0.001) and endometritis (10.5% vs. 3.7%), and a lower prevalence of endometrial polyps (6.1% vs. 14.9%, p=0.01). Congenital malformations were present in 18/114 patients (15.9%), of which 11 cases (9.7%) were a septate/bicornuate uterus. There was no difference between the prevalence of hysteroscopic alterations in patients with two versus three or more miscarriages (35.3% versus 42.8%, χ2=0.395, p=0.529). ConclusionThe frequency of uterine malformations and endometrial disorders can be high in patients with RPL, including those with only two previous miscarriages. This finding supports the investigation of uterine disorders in women with a history of RPL.

HSP70 regulates lipid metabolism of decidual macrophages to maintain normal pregnancy

Dysfunction of decidual macrophages (dMs) are closely associated with recurrent pregnancy loss (RPL) which brings great suffering to patients. Metabolism is essential for regulating macrophage function. Identifying molecules that regulate metabolism and function of dMs is important to revealing the pathogenesis of RPL. Single-cell sequencing data of decidual immune cells from control and RPL patients were downloaded from the GSA database and converted into feature-barcode matrices by Cell Ranger. After quality control, removal of double cell and clustering of all cells, 3579 macrophages were extracted for normalisation, scaling and re-clustering. Function and metabolism analyses were performed by R packages AddMoudleScore, scMetabolism and AUCell. Metabolism clustering based on metabolism-related genes to clarify the metabolic characteristics of macrophages clusters. These results indicated that macrophage characterised by lipid metabolism were reduced in RPL and differential expression genes analysis found that HSP70 was significantly decreased in the RPL group. Furthermore, immunofluorescence staining demonstrated that HSP70 was significantly downregulated in dMs of RPL patients compared to controls. In conclusion, HSP70 may maintain normal pregnancy by regulating lipid metabolism of dMs. This study provides new insights into the molecular mechanisms regulating the function of dMs and provides a theoretical basis for the development of new therapies for RPL.

Association of cytotoxic T-lymphocyte-associated protein 4 polymorphisms with recurrent pregnancy loss: A case-control study.

A large proportion of cases of recurrent pregnancy loss (RPL) are associated with immunological factors. This study investigated the association between single nucleotide polymorphisms of cytotoxic T-lymphocyte-associated protein (CTLA)-4 gene in women with a history of RPL compared to healthy women. A case-control study was performed on 2 groups consisting of 120 healthy women with no history of abortion and at least one delivery (control) and 120 women with a history of 2 or more primary RPLs (case). In addition, 5 mL of peripheral blood sample was taken from all subjects. The frequencies of CTLA-4 rs3087243 and rs231775 polymorphisms were assayed by restriction fragment length polymorphism polymerase chain reaction and rs5742909 using the high-resolution melting real-time polymerase chain reaction method. The mean age of the women in the control and RPL groups were 30.03 4.23 (range 21-37), and 28.64 3.61 yr (range 20-35), respectively. Pregnancy loss numbers ranged between 2-6 in women with a history of RPL, and between 1 and 4 in the successful pregnancy group. Statistical analysis showed a significant difference between the genotypes of GG and AG in the 2 groups in rs3087243 polymorphism (OR 1.00 for GG genotype and OR 2.87 for AG genotype, p = 0.0043). No significant difference was observed in the genotype frequencies of rs231775 and rs5742909 polymorphisms, of the 2 groups (p = 0.37, and p = 0.095), respectively. Our findings indicated that CTLA-4 polymorphism, rs3087243, might be associated with a risk of RPL in Iranian women.

Comparison of Blood Homocysteine Levels between Women with Recurrent Pregnancy Loss and Women with Normal Fertility

Background: Recurrent early pregnancy loss is a common but frequently unexplained obstetric problem. As it happens in early weeks of pregnancy, abnormal vasculogenesis and disordered cell multiplication are possible causes. Hyperhomocysteinemia is associated with both these pathological processes. Objective: To assess the association between hyperhomocysteinemia and recurrent pregnancy loss. Materials and Methods: This case-control observational study was conducted in the Department of Gynaecology and Obstetrics of Bangabandhu Sheikh Mujib Medical University, Dhaka during January to December 2015. Sixty patients were divided into two groups: 30 with recurrent pregnancy loss (RPL) as cases and 30 normal healthy mothers as controls. Results: All the subjects were matched in terms of age (p = 0.504). All were within normal BMI range (19.5 to 25 kg/square meter). Subjects in both groups were comparable in height, weight and BMI. 33.3% women of control group were in the low income stratum (monthly income 20000 taka or less) compared to50% of the RPL group. Control group had an equal mix of educational level from below Secondary School to Post-Graduates whereas there were fewer Post Graduate patients (3.3%) in the RPL group compared to control group (23.3%). The frequency of hyperhomocysteinemia (&gt;15 micromoles/L) was significantly higher in cases compared to controls (46.7 vs.16.7%, p= 0.012). Mean homocysteine level was also significantly higher in cases compared to controls (13.67+/-4.80 vs. 9.87+/-4.84 micromole/L; p= 0.003). Conclusion: This study shows that blood homocysteine level in recurrent pregnancy loss patients is significantly higher compared to normal fertile mothers. There is a strong association between hyperhomocysteinemia and RPL. J Enam Med Col 2021; 11(1): 10-17

Increased levels of nerve growth factor accompany oxidative load in recurrent pregnancy loss. Machine learning applied to FT-Raman spectra study.

The presented article is focused on developing and validating an efficient, credible, minimally invasive technique based on spectral signatures of blood serum samples in patients with diagnosed recurrent pregnancy loss (RPL) versus healthy individuals who were followed at the Gynecology department. A total of 120 participants, RPL disease (n = 60) and healthy individuals (n = 60), participated in the study. First, we investigated the effect of circulating nerve growth factor (NGF) in RPL and healthy groups. To show NGF's effect, we measured the level of oxidative loads such as Total Antioxidant Level (TAS), Total Oxidant Level (TOS), and Oxidative Stress Index (OSI) with Beckman Coulter AU system and biochemical assays. We find a correlation between oxidative load and NGF level. Oxidative load mainly causes structural changes in the blood. Therefore, we obtained Raman measurements of the participant's serum. Then we selected two Raman regions, 800 and 1800cm-1, and between 2700cm-1 and 3000cm-1, to see chemical changes. We noted that Raman spectra obtained for RPL and healthy women differed. The findings confirm that the imbalance between reactive oxygen species and antioxidants has important implications for the pathogenesis of RPL and that NGF levels accompany the level of oxidative load in the RPL state. Biomolecular structure and composition were determined using Raman spectroscopy and machine learning methods, and the correlation of these parameters was studied alongside machine learning technologies to advance toward clinical translation. Here we determined and validated the development of instrumentation for the Analysis of RPL patients' serum that can differentiate from control individuals with an accuracy of 100% using the Raman region corresponding to structural changes. Furthermore, this study found a correlation between traditional biochemical parameters and Raman data. This suggests that Raman spectroscopy is a sensitive tool for detecting biochemical changes in serum caused by RPL or other diseases.

Depression, Anxiety, and Stress in Kazakhstani Women with Recurrent Pregnancy Loss: A Case-Control Study.

Background: Recurrent pregnancy loss (RPL) is associated with increased incidence and severity of depression, anxiety, and stress, and screening for these comorbidities following miscarriages is beneficial for women with RPL who are planning future pregnancies. This study aims to investigate depression, anxiety, and stress among Kazakhstani women with RPL. Methods: This was a case−control study involving 70 women with confirmed RPL and 78 ethnically matched control women. Depression, anxiety, and stress were evaluated using the Depression Anxiety Stress Scales (DASS)-21 instrument. Linear regression and correlation analysis were used in assessing the association of RPL with symptoms of depression, and/or anxiety, and/or stress, after adjusting for key covariates. Results: Women with RPL were found to have significantly higher mean scores for depression (p < 0.001), anxiety (p < 0.001), and stress (p < 0.001) symptoms. Mild−moderate stress and mild−moderate and severe−extreme depression and anxiety symptoms were more frequent in the RPL group than in the control group. Regression analysis demonstrated that RPL was the only significant variable associated with anxiety, depression, and stress symptoms. Conclusion: The results of this study suggest that women with RPL are more likely to experience heightened symptoms of depression, anxiety, and stress. Proper psychological counseling is recommended for women with RPL, as well as their spouses.

Cytogenetics evaluation of 261 couples with first-trimester recurrent pregnancy loss: A prevalent case–control study

Introduction: Recurrent pregnancy loss (RPL) is a common occurrence which affects approximately 15-20% of couples. Chromosomal abnormality is an important cause of recurrent abortions especially if either of the partner is a carrier of balanced translocation. The current study aimed to determine the prevalence of chromosomal abnormalities in couples suffering from first trimester RPL and compare with normal control. Materials and Methods: A prospective case-control study, in which 261 couples with history of two or more abortions were evaluated for various chromosomal abnormalities; and compared with 190 healthy couples with no history of abortion and at least one normal biological child. Peripheral blood T-lymphocytes were cultured using RPMI-1640 medium for obtaining metaphases and chromosomal analysis. SPSS software and Student's t test were used. p value < 0.05 was considered statistically significant. Results: Among 261 couples in RPL group, 240(91.95%) had normal karyotype, 17(6.51%) had major chromosomal abnormalities and 04(1.53%) had polymorphic variants. Most of the couples had two abortions (39.8%). Females were more commonly affected with M:F=0.214. Structural abnormalities (n=12,70.59%) were more frequent than numerical abnormalities (n=5,29.41%). There was no statistical correlation between age, number of abortions and major chromosomal abnormalities (p=0.06). Conclusion: Chromosomal aberrations in carrier parents, predispose them to RPL and can also be transmitted to the offspring which may results in imbalance in their genetic constitution, thus justifying the requirement of cytogenetic testing in these patients.

Role of Sperm DNA Fragmentation Index in Semen Analysis in Couples with Unexplained Recurrent Pregnancy Loss: A Case-control Study

Introduction: Infertility has been on the rise, and male factor infertility has gained attention in cases of Recurrent Pregnancy Loss (RPL). Traditional Semen Analysis (SA) has been the main diagnostic tool, but with advancements in Assisted Reproductive Technology (ART) and the need for more accurate testing, there is a demand for improved diagnostic tests that correlate with reproductive outcomes. Sperm DNA Fragmentation Index (DFI) is a parameter used to assess the degree of sperm DNA damage and is considered crucial in evaluating semen quality. Aim: To investigate the role of Sperm DNA Fragmentation (SDF) in patients presenting with RPL compared to a control group. Materials and Methods: The study was conducted over a two-year period from January 2021 to December 2022 at the Centre for Infertility Management and Assisted Reproduction (CIMAR), Edappal Hospital, Edappal, Kerala, India. The control group (Group A; n=31) comprised males aged between 21-45 years whose partners had no history of recurrent abortions. The case group (Group B; n=31) included males with two or more pregnancy losses. Sperm DFI analysis and routine SA were performed in both groups to assess semen parameters such as volume, concentration, progressive and non progessive motility, morphology, and DFI. Statistical analysis was performed using the Independent student t-test. Results: In the present study the mean age of males in Group A was 37.03±5.416 years and in Group B was 35.44±4.552 years. There were no significant differences observed between the case and control groups in terms of sperm volume (p=0.301), concentration (p=0.155), progressive motility (p=0.207), non progessive motility (p=0.178), and morphology (p=0.362). However, a statistically significant difference was found between the RPL and control groups for DFI (p&lt;0.001), with a mean value of ±8.15 in the control group and ±19.35 in the case group. Conclusion: The present study demonstrates that SDF is an important factor in RPL, with couples experiencing RPL showing a higher incidence of SDF. Therefore, incorporating SDF analysis alongside routine SA should be considered, particularly in patients with a history of RPL.

Self-Reported Depression among Chinese Women with Recurrent Pregnancy Loss: Focusing on Associated Risk Factors.

(1) Background: To investigate whether women suffering from recurrent pregnancy loss (RPL) have a higher prevalence of self-reported depression than healthy controls and to assess the associated risk factors for RPL women self-reporting the symptom of depression. (2) Methods: A cross-sectional study investigating 247 women with histories of RPL and 193 healthy women was performed in Southwest China. The Zung Self-Rating Depression Scale (SDS) was used to measure self-reported depression, and the prevalence of self-reported depression was compared between the two groups. Sociodemographic data for the two groups and clinical information for the RPL group were collected by questionnaires. (3) Results: The prevalence of self-reported depression was higher in the RPL group than in the control group (45.3% vs. 30.1%, p &lt; 0.01). Subgroup analyses indicated that the statistical difference in the prevalence of self-reported depression was significant in the subgroups of women in the first trimester of gestation, age ≥ 36 years, BMI ≥ 18.5 kg/m2, working hours ≤ 8 h/day, university and higher education, and urban residence. Multivariable logistic analysis indicated that age ≥ 36 years, &gt;2 times of spontaneous miscarriages, and no history of live birth were independent risk factors for RPL women self-reporting depression. (4) Conclusions: A higher prevalence of self-reported depression was observed in RPL patients than in healthy women. The psychological status for RPL patients with age ≥ 36 years, &gt;2 times of spontaneous miscarriages, or without a history of a live birth needs to be further addressed.

Investigation of the role of X chromosome inactivation and androgen receptor CAG repeat polymorphisms in patients with recurrent pregnancy loss: a prospective case–control study

BackgroundPrevious research has revealed that skewed X chromosome inactivation (SXCI) and androgen receptor (AR) CAG polymorphisms are associated with increased risk of recurrent pregnancy loss (RPL); however, the results are conflicting, and the underlying mechanisms remain unclear. This study investigated the role of SXCI and AR CAG polymorphisms in patients with RPL and explored whether the underlying mechanisms were related to the ovarian reserve and preimplantation embryo aneuploidy.MethodsThis was a prospective case-control study carried out in a tertiary hospital-based reproductive medicine center. An external validation RPL cohort was recruited during the study period. Data on baseline and cycle characteristics were collected. X-chromosome inactivation (XCI) was measured using a human AR assay. AR polymorphisms were assessed using quantitative fluorescent polymerase chain reactions and direct sequencing. Blastocysts of the patients with RPL were tested by single nucleotide polymorphism microarray based preimplantation genetic testing for aneuploidy.ResultsIn total, 131 patients with idiopathic RPL and 126 controls were included for the case-control study. Patients with RPL exhibited a significantly more skewed XCI distribution pattern (67.71 ± 10.50 vs. 64.22 ± 10.62, p = 0.011), as well as significantly shorter bi-allelic mean (18.56 ± 1.97 vs. 19.34 ± 2.38, p = 0.005) and X-weighted bi-allelic mean (18.46 ± 2.02 vs. 19.38 ± 2.53, p = 0.001) of AR CAG repeats. Multivariate logistic regression models indicated that CAG repeat < 20, SXCI, and duration of stimulation were independently associated with the risk of RPL. However, SXCI and AR CAG polymorphisms were not associated with ovarian reserve or preimplantation embryo aneuploidy in the RPL group, and the same results were attained in a separate validation cohort of 363 patients with RPL.ConclusionSXCI and AR CAG polymorphisms are related to RPL; however, these two factors do not lead to RPL by affecting the ovarian reserve or increasing embryo aneuploidy. The roles of SXCI and AR CAG in RPL may involve other mechanisms that require further investigation.Trial registration:NCT02504281, https://www.clinicaltrials.gov (Date of registration, 21/07/2015; date of enrolment of the first subject, 30/07/2015).

Does a history of recurrent pregnancy loss affect subsequent obstetric outcomes and placental findings in in vitro fertilization?

To assess perinatal outcomes and placental findings in in vitro fertilization (IVF) patients with previous recurrent pregnancy loss (RPL). This was a retrospective cohort of live singleton births following IVF at a single university-affiliated center between 2009 and 2017. Outcomes were compared between patients with previous RPL, defined as two miscarriages (RPL group), and patients without RPL (control group). Placental examination was performed for all deliveries irrelevant of complications, and findings categorized according to the Amsterdam Placental Workshop Consensus. One hundred seventy-two deliveries of women with previous RPL (RPL group) were compared to 885 controls. Maternal age, 36.2 ± 4.2 vs. 35.4 ± 4.2years, p = 0.02, and rate of uterine fibroids, 12.7% vs. 7.3%, p = 0.01, were higher in the RPL group. The rate of nulliparity was lower in the RPL group, 63.3% vs. 74.1%, p = 0.003. Unexplained infertility and diminished ovarian reserve were more common in the RPL group and male factor infertility in controls. There was a lower rate of fresh embryo transfer in the RPL group, 50.5% vs. 64.7%, p < 0.001, and reduced endometrial thickness. Similar obstetric outcomes were noted in the groups after adjustment for confounders. Placental examinations were notable for lower placental thickness, β - 0.17cm, 95%CI - 0.30-(- 0.0), a lower rate of velamentous cord insertion, aOR 0.44, 95%CI 0.20-0.95, and a higher rate of villous infarction, aOR 2.82, 95%CI 1.28-6.20 in the RPL group. In IVF patients with a history of RPL, subsequent deliveries were associated with a limited number of placental lesions, yet with similar obstetric outcomes.

Microbial Diversity and Pathogenic Properties of Microbiota Associated with Aerobic Vaginitis in Women with Recurrent Pregnancy Loss.

Recurrent pregnancy loss (RPL) is a major reproductive problem that affects approximately 5% of couples. The objective of this study was to assess vaginal flora dysbiosis in women suffering from unexplained RPL and to investigate the pathogenic properties of the microbiota associated with aerobic vaginitis (AV). The study included one hundred fifteen women, 65 with RPL and 50 controls. The diversity of vaginal microbiota isolated was evaluated by molecular sequencing. Then, pathogenic factors, such as acid-resistance, antibiotics susceptibility, and biofilm formation were evaluated. The prevalence of AV was five-fold higher in the RPL group than in the controls (64.6% vs. 12.0%). The most prevalent isolates in the case group were Enterococcus spp. (52%) and Staphylococcus spp. (26%). All bacterial strains tolerate low pH. The prevalence of multidrug resistance (MDR) among all bacteria was 47.7%. Of all strains, 91.0% were biofilm producers. The presence of MDR was found to be related to biofilm formation. The results provide evidence supporting an increased presence of dysbiosis of the vaginal flora, especially AV, in women with RPL in Tunisia. The viability of the AV-associated bacteria and their persistence in the genitals may be due to their ability to resist low pH and to produce a biofilm.

A unique investigation of thallium, tellurium, osmium, and other heavy metals in recurrent pregnancy loss: A novel approach.

To study the impact of heavy metals especially tellurium, thallium, and osmium, in recurrent pregnancy loss (RPL) and to study their association with antioxidant status and DNA damage. This case-control study included women with RPL (n= 30) and healthy pregnant women as control (n= 30). Following blood collection, serum levels of thallium, tellurium, osmium, lead, mercury, and cadmium were estimated by inductively coupled plasma mass spectrophotometer. Women with RPL exhibited significantly higher levels of heavy metals (P< 0.001) when compared with control women. Intriguingly, increased levels of serum thallium, tellurium, osmium, and lead were negatively correlated with total antioxidant status (P< 0.05). Further, the RPL group demonstrated strong positive correlation between heavy metals (thallium, tellurium, osmium, lead) and DNA damage (P< 0.05). No significant correlation between other heavy metals and markers of cellular damage was noted. Enhanced levels of heavy metals in women with RPL and correlation of thallium, tellurium, osmium, and lead with markers of cellular damage reflect the role of heavy metal poisoning, especially thallium, tellurium, and osmium, as potential risk factor in the etiology underlying recurrent miscarriage.