Inflammatory shift alterations of proinflammatory and anti-inflammatory cytokines in unexplained recurrent miscarriage patients.

This study aimed to investigate the pro-inflammatory and anti-inflammatory cytokines status in the peripheral blood of uRM patients. The plasma pro-inflammatory (IFN-γ, IL-6, IL-1ß, and TNF-α) and anti-inflammatory (TGF-ß1, IL-10, and IL-4) cytokines of 25 patients with uRM were compared to 33 women with a successful pregnancy. It was concluded that patients with uRM have an excess pro-inflammatory cytokines status.

Intrauterine perfusion of dexamethasone improves pregnancy outcomes in recurrent reproductive failure patients with elevated uterine natural killer cells. A retrospective cohort study.

OBJECTIVE: To determine the effect of intrauterine perfusion of dexamethasone (DXM) on pregnancy outcomes in recurrent reproductive failure (RRF) patients with elevated uNK cells. METHODS: This retrospective cohort study included 132 RRF patients with elevated uNK cells: 56 patients received DXM treatment and 76 patients refused it in the frozen-thawed embryo transfer cycles. To determine the efficacy of intrauterine perfusion of DXM, multivariate logistic regression models and diagnosis-based subgroup analysis were performed. We also compared the pregnancy outcomes of patients with different responsiveness to DXM treatment. RESULTS: Intrauterine perfusion of DXM significantly improved clinical pregnancy rate (aOR: 3.188, 95% CI: 1.395-7.282, P = .006) and live birth rate (aOR: 3.176, 95% CI: 1.318-7.656, P = .010) in RRF patients with elevated uNK cells, but there was no significant association with miscarriage rate. Subgroup analysis revealed that intrauterine perfusion of DXM in patients with recurrent implantation failure (RIF) showed significant improvement in clinical pregnancy rate (aOR: 6.110, 95% CI: 1.511-24.713, P = .011) and live birth rate (aOR: 9.904, 95% CI: 1.963-49.968, P = .005), but there was insufficient evidence of benefit in recurrent pregnancy loss (RPL) patients. Additionally, uNK cell levels dropped to normal range was achieved in only 35.90% of RRF patients after DXM treatment, no significant difference was found in pregnancy outcomes among patients with different responsiveness to DXM treatment (all P > .05). CONCLUSION: Intrauterine perfusion of DXM was a promising and effective treatment to enhance clinical pregnancy rate and live birth rate in RRF women with abnormally elevated uNK cells, and RIF patients are more likely to benefit than RPL patients.

Platform for Quantitative Detection of Endometrial Immune Cells Based on Immunohistochemistry and Digital Image Analysis.

To evaluate the endometrial immune microenvironment of patients with recurrent miscarriage (RM), a digital immunohistochemistry image analysis platform was developed and validated to quantitatively analyze endometrial immune cells during the mid-luteal phase. All endometrium samples were collected during the mid-luteal phase of the menstrual cycle. Paraffin-embedded endometrial tissues were sectioned into 4 µm thick slides, and immunohistochemistry (IHC)staining was carried out for detecting endometrial immune cells, including CD56+ uNK cells, Foxp3+ Tregs, CD163+ M2 macrophages, CD1a+ DCs, and CD8+ T cells. The panoramic slides were scanned using a digital slide scanner and a commercial image analysis system was used for quantitative analysis. The percentage of endometrial immune cells was calculated by dividing the number of immune cells in the total endometrial cells. Using the commercial image analysis system, quantitative evaluation of endometrial immune cells, which are difficult or impossible to analyze with conventional image analysis, could be easily, and accurately analyzed. This methodology can be applied to quantitatively characterize the endometrium microenvironment, including interaction between immune cells, and its heterogeneity for different reproductive failure patients. The platform for quantitative evaluation of endometrial immune cells may be of important clinical significance for the diagnosis and treatment of RM patients.

The effectiveness and safety of intrauterine infusion of autologous regulatory T cells (Tregs) in patients with recurrent pregnancy loss and low levels of endometrial FoxP3+ cells: A retrospective cohort study.

PROBLEM: Regulatory T cells (Tregs) are a specialized type of T cells that help maintain immune tolerance and homeostasis. The potential of Tregs cell-based therapies in treating diseases has been demonstrated in several clinical trials, which have shown promising outcomes and high safety in autoimmune diseases, transplant rejection, and graft-versus-host disease. However, their effectiveness and safety in improving endometrial receptivity and reducing pregnancy loss in human reproduction are unknown. METHOD OF STUDY: The study used a retrospective design and included patients with recurrent pregnancy loss (RPL) and lower levels of endometrial FoxP3+ Tregs. Patients in the Tregs group (n = 33) received intrauterine Tregs infusion three times during the follicular phase, while the control group (n = 28) did not receive any intrauterine infusion. RESULTS: The intrauterine infusion of autologous Tregs increased the levels of FoxP3+ Tregs and CD56+ NK cells. Patients in the Treg group had higher live birth rates and lower miscarriage rates, especially early miscarriage rates. However, the two groups had no differences in the implantation rate, clinical pregnancy rate, and percentage of preterm delivery. CONCLUSIONS: The findings suggest that intrauterine Tregs infusion may be a potential therapeutic approach for RPL. Further research in larger clinical trials is needed to confirm these findings.

γδ-T cell with high toxic potential was associated with recurrent miscarriage.

PROBLEM: RM is a common clinical disease in reproduction, affecting approximately 1%-3% of women worldwide. Previous studies have shown the role of peripheral blood γδ-T cells during physiological pregnancy. However, the relationship between the immune status of peripheral blood γδ-T cells and RM is still not well defined. METHOD OF STUDY: In this study, mid-luteal peripheral blood from 51 RM patients and 40 healthy women was collected to determine the immune status of γδ-T cells. The percentage of peripheral blood γδ-T cells, and the molecules mediating their toxic potential, including cytotoxic granules (perforin, granzyme B, and granulysin) and receptors (NKG2D, CD158a, and CD158b), were detected by flow cytometry. RESULTS: Compared to healthy control, an increase in the proportion of total CD3+ T cells in lymphocytes and a decrease in the ratio of γδ-T cells to CD3+ T cells were observed in patients with RM. The percentages of granzyme B+ γδ-T cells and CD158a+ γδ-T cells in total γδ-T cells or lymphocytes were significantly increased in patients with RM, compared with healthy control. Conversely, CD158b+ γδ-T cells in total γδ-T cells or lymphocytes were significantly decreased in the RM group. CONCLUSION: Increased peripheral blood γδ-T cell with high toxic potential was associated with RM.

Comparing the peri-implantation endometrial T-bet/GATA3 ratio between control fertile women and patients with recurrent miscarriage: establishment and application of a reference range.

STUDY QUESTION: Is the ratio of endometrial T-box expressed in T cell (T-bet) and GATA-binding protein 3 (GATA3) changed in patients with recurrent miscarriage (RM) compared to fertile controls? SUMMARY ANSWER: Our study showed a significantly higher T-bet/GATA3 ratio in patients with RM compared with fertile controls. WHAT IS KNOWN ALREADY: The endometrial T-bet (Th1 lineage-committed transcription factor)/GATA3 (Th2 lineage-committed transcription factor) ratio could represent the Th1/Th2 balance, which is particularly important for healthy pregnancy. However, a reliable reference range for the T-bet/GATA3 ratio during the peri-implantation period has not yet been established for use in clinical practice. STUDY DESIGN, SIZE, DURATION: This was a retrospective study carried out in a private fertility center. The control group included 120 women in couples undergoing IVF treatment for male infertility, who had experienced a live-birth baby following the first IVF cycle. The study group included 93 women diagnosed with RM that experienced at least two consecutive clinically spontaneous miscarriages before gestational week 12. The ratio of T-bet/GATA3 was calculated in the control group and RM group. PARTICIPANTS/MATERIALS, SETTING, METHODS: Endometrium samples were collected at mid-luteal phase of the menstrual cycle prior to IVF treatment or pregnancy. The percentage of T-bet+ and GATA3+ cells in total endometrial cells was analyzed using immunohistochemical staining and quantitative analysis. MAIN RESULTS AND THE ROLE OF CHANCE: Using the 95th percentile to define the upper limits of the endometrial T-bet/GATA3 ratio during the mid-luteal phase, the reference range of control fertile women was ≤0.22. Compared with the control group, the RM group exhibited a significantly higher T-bet/GATA3 ratio (P = 0.02), and 19.4% (18/93) women with RM exhibited a T-bet/GATA3 ratio above the reference range in the mid-luteal phase. LIMITATIONS, REASONS FOR CAUTION: All patients were recruited from a single center. The stability and clinical value of the endometrial T-bet/GATA3 ratio require further investigation. WIDER IMPLICATIONS OF THE FINDINGS: The present study suggests that an abnormal endometrial T-bet/GATA3 ratio may be one of the risk factors of RM. Further studies are needed to follow up the pregnancy outcomes in patients with RM with normal and abnormal endometrial T-bet/GATA3 ratio according to the reference range. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by Shenzhen Fundamental Research Program (JCYJ20180228164631121, JCYJ20190813161203606, JCYJ20220530172817039). There are no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Impact of body mass index on peripheral and uterine immune status in the window of implantation in patients with recurrent reproductive failure.

This study aimed to investigate whether maternal obesity affects the immune status of peripheral blood and endometrium in patients with recurrent reproductive failure classified according to their body mass index (BMI). A total of 228 repeated implantation failure (RIF) and 266 recurrent miscarriage (RM) patients were enrolled in the study and further subdivided into three groups according to their BMI: (i) normal weight (18.5≤ BMI <23); (ii) overweight (23≤ BMI <25); and (iii) obese (BMI ≥25). Peripheral blood and endometrium samples were collected in the mid-luteal phase before IVF treatment or natural pregnancy. Peripheral immunocytes were analyzed by flow cytometry, while uterine immune cells were subjected to immunohistochemistry. In RM patients, significantly increased peripheral helper T cells and decreased cytotoxic T cells, NK cells were observed in the obese group compared with the normal-weight group. Meanwhile, in the endometrium, the percentage of NK cell, macrophage cell, M2 macrophage cell, and Treg cell significantly reduced with increased BMI in RIF patients, and the percentage of NK cell and M2 macrophage cell significantly decreased with increased BMI in RM patients. In conclusion, obesity may cause endometrial immune disorder in recurrent reproductive failure women, but was only associated with the peripheral immune change in RM patients.

Establishment of reference intervals of endometrial immune cells during the mid-luteal phase.

This study aimed to develop reference intervals (RIs) of endometrial immune cells in control infertile women during the mid-luteal phase, and compare with the proportion of endometrial immune cells in recurrent reproductive failure (RRF) patients. Endometrial tissue sections were obtained from 113 fertile women and 79 patients with RRF, including 40 patients who had suffered recurrent miscarriage (RM) and 39 patients with repeated implantation failure (RIF) during the mid-luteal phase of the menstrual cycle. Immunohistochemical staining and quantitative analysis of CD56+, Foxp3+, CD163+, CD1a+ and CD8+ cells were performed in endometriums. RIs of endometrial immune cells in control infertile women were as follows: CD56+ uterine natural killer cells (uNK) cells, 1.785-8.712%, forkhead box P3 (Foxp3)+ Tregs, 0.041-0.154%, CD163+ M2 macrophages, 0.298-1.492%, CD1a+ dendritic cells (DCs), 0.006-0.081% and CD8+ T cells, 0.674-2.504%. Compared with control infertile women, the percentage of endometrial CD56+ uNK cells, CD163+ M2 macrophages, CD1a+ DCs and CD8+ T cells were significantly increased in patients with RRF. Moreover, Foxp3+ Tregs levels were decreased in patients with RRF, and were statistically significant only in patients with RM. In conclusion, the RIs of endometrial immune cells were established in control infertile women during the mid-luteal phase, and a disordered endometrial immune microenvironment was observed in patients with RRF. The RIs of endometrial immune cells may be of important clinical significance for the treatment of RRF.

Chronic endometritis impairs embryo implantation in patients with repeated implantation failure: A retrospective study.

OBJECTIVE: To assess whether chronic endometritis (CE) affects embryo implantation in patients with repeated implantation failure (RIF). MATERIALS AND METHODS: We retrospectively analyzed 126 RIF patients who were never diagnosed with CE and received no prior antibiotic therapy. Endometrial specimens obtained by endometrial scratching during mid-luteal phase were immunostained by CD138, a hallmark plasmacyte marker, to identify CE. Pregnancy outcome in RIF patients who underwent IVF-ET frozen-embryo within transfers 6 months after endometrial scratching was compared between women with and without CE. RESULTS: The prevalence of CE in patients with RIF was found to be 11.9% (15/126). Moreover, a significantly reduced clinical pregnancy rate was observed in RIF patients with CE (20% vs. 46.85%; p = 0.04). The live birth rate also exhibited a decreasing trend in RIF patients with CE, although there was no statistically significant difference (20% vs. 41.58%; p = 0.109). CONCLUSIONS: CE may be involved in the failure of embryo implantation and reduced clinical pregnancy outcome in patients with RIF.

Intrauterine infusion of human chorionic gonadotropin improves the endometrial FoxP3+ Tregs level and pregnancy outcomes in patients with lower endometrial FoxP3+ Tregs.

Intrauterine infusion of human chorionic gonadotropin (hCG) is suggested to have the capacity to recruit regulatory T cells (Tregs) in the endometrium. However, the pregnancy outcome for infertile women with a lower level of Tregs after hCG intrauterine infusion remains unknown. By examining the expression of FoxP3+ Tregs in the endometrium, this study aimed to evaluate the effectiveness of hCG intrauterine infusion in infertile women with lower endometrial Tregs in improving the Tregs level and the pregnancy outcome. This cohort study included 150 women aged 38 and younger with a lower FoxP3+ Tregs level in the mid-luteal phase. Patients were divided into the control group (n = 73), and hCG group (n = 77). Patients in the hCG group received three times of hCG intrauterine infusion during the follicular phase in the next biopsy and the embryo transfer cycles. The results showed that the endometrial FoxP3+ Tregs level increased significantly after hCG intrauterine infusion (P < 0.001). The effective rate in rescuing the endometrial Tregs level was 77.92% (60/77). The clinical pregnancy rate was increased significantly in the hCG group than the control group (54.8% vs. 74.0%, P = 0.014). The logistic regression result showed that hCG intrauterine infusion [adjusted odds ratio (aOR) (95% confidence interval (CI)) = 2.347 (1.119, 4.923), P = 0.024] and blastocyst transfer [aOR (CI) = 2.630 (1.090, 6.346)] are two independent factors contributing to the improved pregnancy outcome. These data highlighted the immune regulatory role of hCG intrauterine infusion and might facilitate the personal immunotherapy progress for unexplained infertility in patients with a lower endometrial Tregs level.

Transcriptomic Analysis Reveals Endometrial Dynamics in Normoweight and Overweight/Obese Polycystic Ovary Syndrome Women.

The aim of this work was to identify the transcriptomic characteristics of the endometrium in normoweight and overweight/obese polycystic ovary syndrome (PCOS) potentially underlying the pathogenesis. This study included 38 patients undergoing in vitro fertilization: 22 women with PCOS and 16 matched controls. Each of the groups was subdivided into normoweight (body mass index (BMI) < 25 kg/m2) and overweight/obese (BMI ≥25 kg/m2) subgroups. Endometrium samples were collected in the secretory phase from controls or in a modeled secretory phase using daily administration of progesterone from women with PCOS before in vitro fertilization treatment. Transcriptome profiles were assessed by high-throughput RNA sequencing to investigate distinct endometrial gene expression patterns in PCOS. Bioinformatics analyses revealed that the endometrium from PCOS expresses significantly different transcripts encoding endometrial receptivity, inflammatory response, angiogenesis, and energy metabolism. Additionally, our study demonstrated that the differentially expressed genes between normoweight and overweight/obese PCOS are involved in fatty acid metabolism, endometrial decidualization, and immune response. For the first time, we have described the transcriptome characteristics of normoweight and overweight/obese PCOS endometria. Our results indicate different endometrial gene expressions between different subtypes of PCOS and non-PCOS women, which might affect endometrial functions in PCOS patients.

Predictive role of endometrial T-bet/GATA3 ratio during mid-luteal phase for live birth in patients undergoing in vitro fertilization: A retrospective observational study.

OBJECTIVES: To investigate whether endometrial T-bet (Th1 lineage-committed transcription factor)/GATA3 (Th2 lineage-committed transcription factor) ratio has predictive potential for embryo implantation in infertile women undergoing in vitro fertilization-embryo transfer (IVF-ET). STUDY DESIGN: We performed a retrospective observational study. In total, this study included 319 infertile women (253 women as the development cohort and 66 women as the validation cohort). Samples were obtained by endometrial scratching in the mid-luteal phase before IVF-ET treatment. MAIN OUTCOME MEASURES: Immunohistochemistry was utilized to analyze the expression levels of T-bet and GATA3 in the endometrium. Predictive value of endometrial T-bet/GATA3 for live birth were analyzed. RESULTS AND CONCLUSIONS: In the development cohort, the T-bet/GATA3 ratio was significantly lower in women with live birth than those patients with non-live birth [0.148 (0.101, 0.212) vs. 0.246 (0.170, 0.399), P＜0.0001]. In the validation cohort, changes in endometrial T-bet/GATA3 were similar among these groups. The endometrial T-bet/GATA3 ratio was an independent predictor of live birth after correction for patient age, anti-Mullerian hormone (AMH), quality of embryos transferred and other clinical characteristics (aOR = 0.280, 95 % CI: 0.169-0.462, P＜0.001). We developed and validated that an endometrial T-bet/GATA3 ratio at the cut-off of 0.22 had significant predictive value for live birth (developmental cohort: AUC = 0.76, 95 % CI: 0.70-0.81, P < 0.0001. validation cohort: AUC = 0.85 95 % CI: 0.76-0.95, P < 0.0001). Our results suggest that elevated endometrial T-bet/GATA3 ratio is an independent marker of live birth in infertile patients.

A History of Endometriosis Is Associated With Decreased Peripheral NK Cytotoxicity and Increased Infiltration of Uterine CD68+ Macrophages.

Women with endometriosis may have a defective immune system. However, evidence of the immune responses of endometriosis patients with a history of endometriosis surgery is lacking, and the association between the location of endometriosis lesions and immune responses is unclear. This retrospective study included 117 females with reproductive failure and a history of endometriosis and 200 females with reproductive failure but without endometriosis to analyze their endometrial and peripheral immune responses. The results show that endometriosis was associated with decreased peripheral natural killer (NK) cytotoxicity and increased uterine macrophages. Peripheral NK cytotoxicity at effector-to-target ratios of 25:1 and 50:1 was significantly reduced in women with a history of endometriosis from that of the control group (26.6% versus 33.3% and 36.1% versus 43.3%, respectively, both P < 0.001). Furthermore, after further division of patients into three subgroups according to the location of endometriosis lesions, we observed that NK cytotoxicity in the endometriosis subgroups, especially the mixed endometriosis group, was strongly decreased from that of the controls (P = 0.001). The endometrial CD68+ macrophage proportion in the mixed endometriosis subgroup was higher than that in the control group (2.8% versus 2.1%, P = 0.043). In addition, the baseline estradiol (E2) level was weakly correlated with the percentage of endometrial macrophages (r = 0.251, P = 0.009), indicating a potential association among the endocrine system, endometrial immune environment, and endometriosis. This study indicated that peripheral NK cytotoxicity and endometrial immune cell profiles could be useful for diagnosing and treating endometriosis and endometriosis-related reproductive diseases.

Evaluation of endometrial immune status of polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is described as a low-grade chronic inflammatory state. However, there are limited studies on the specific endometrial immune status of PCOS patients. Whether this endometrial immune cell pattern is intrinsic to PCOS or the consequence of PCOS-associated obesity is a subject of debate. This study retrospectively included one hundred women diagnosed with PCOS and ninety-five normal fertile controls, which further divided into four groups (normoweight PCOS; overweight PCOS; normoweight control; overweight control) based on body mass index. The percentages of endometrial CD68+ macrophages (1.97 % vs. 1.17 %; P < 0.001), CD163+ M2 macrophages (2.30 % vs. 1.83 %; P = 0.001), CD1a+ iDCs (0.044 % vs. 0.029 %; P = 0.002), CD83+ mDCs (1.72 % vs. 1.07 %; P < 0.001) and CD8+ T cells (2.82 % vs. 1.95 %; P < 0.001) were significantly higher in normoweight PCOS women than normoweight controls. The percentage of CD68+ macrophages (2.09 % vs. 1.15 %; P < 0.001) was significantly higher in overweight PCOS women compared with overweight controls. In multivariant linear regression analysis, participants' PCOS status was the main predictors of endometrial CD68+ macrophages, CD163+ M2 macrophages, CD1a+ iDCs, CD83+ mDCs and CD8+ T cells in the whole study population. Additionally, in PCOS group, positive correlations were found between endometrial CD56+ NK, CD163+ M2 macrophages and QUICKI, indicating there was an association between endometrial immune cells and insulin resistance in PCOS women. Our study suggests that women with PCOS have altered endometrial immune cells, which may reflect a state of chronic low grade inflammation. The chronic inflammation, independent of obesity, may help understand the pathophysiologic mechanisms of intrinsic PCOS.

The low cytotoxic activity of peripheral blood NK cells may relate to unexplained recurrent miscarriage.

PROBLEM: Unexplained recurrent miscarriage (uRM) is defined as two or more spontaneous abortions prior to 20 weeks of gestation with unknown etiology. Peripheral blood natural killer (pNK) cells contact with the villus and exert important role in normal pregnancy. However, it is still controversial about the association between pNK cytotoxicity and uRM, and the underlying mechanism remains unknown so far. METHOD OF STUDY: In this study, we aim to compare the percentage, immunophenotype, and function of pNK cells between patients with uRM and fertile controls. The peripheral blood was collected from 49 patients with uRM and 11 fertile women in their middle luteal phase of the menstrual cycle. pNK cells were co-cultured with K562 cells at different cell ratios to measure the cytotoxicity. The percentage of CD3- CD56+ , CD3- CD56bright , and CD3- CD56dim pNK was analyzed by flow cytometry and quantified to evaluate the expression of cytotoxic granules (granzyme B, granulysin, and perforin), and the cell surface receptors related to pNK cell cytotoxicity (NKG2D, NKp30, NKp46, CD158a, and CD158b) were also detected. RESULTS: The general linear model analysis showed that pNK cell cytotoxicity in patients with uRM was significantly lower than that in fertile controls. In addition, the ratios of NKG2D/CD158a, NKp30/CD158a, and NKp46/CD158a in CD3- CD56bright pNK subsets were significantly lower in uRM group than that in fertile control. The logistical regression analysis showed that the reduced NKp30/CD158a, NKp46/CD158a ratios in CD3- CD56bright pNK subsets were significantly associated with uRM. CONCLUSION: Our results suggested that a low pNK cytotoxicity, which is mediated by inhibitory signals, might be associated with uRM.

Association between endometrial indoleamine 2,3-dioxygenase expression level and pregnancy outcomes in women undergoing first in vitro fertilization treatment.

BACKGROUND: Indoleamine 2,3-dioxygenase (IDO) has been reported to play a key role in placental development during normal pregnancy. However, the question of whether endometrial IDO expression affects in vitro fertilization (IVF) pregnancy outcomes remains unclear. The current study was undertaken to investigate whether there was any association between endometrial IDO immunohistochemical staining and IVF treatment outcome. METHODS: This retrospective study was designed to compare pregnancy outcomes among women with different endometrial IDO expression levels under their first IVF treatment. A total of 140 women undergoing their IVF treatment were selected from January 2017 to December 2017. Endometrial samples were collected during mid-luteal phase before IVF cycle. The endometrial IDO expression levels were analyzed by immunohistochemistry, and compared between women who were pregnant or not. A logistic regression analysis was performed to determine the impact of endometrial IDO staining on live birth. RESULTS: No significant differences in the endometrial IDO immunohistochemical staining were found between women who had clinical pregnancy and those who failed (P>0.05). However, the endometrial IDO staining was significantly higher among women who had live birth compared with those who had no live birth (P=0.031). Additionally, after adjusting for differences in maternal age, BMI and duration of gonadotropin stimulation, women with higher IDO expression level had an increased live birth rate (adjusted odds ratio [aOR] 2.863, 95% confidence interval [CI] 1.180-6.947). CONCLUSIONS: Higher endometrial IDO expression level during mid-luteal phase is associated with an increased live birth rate in women undergoing their first IVF treatment.

Diagnosis of chronic endometritis: How many CD138+ cells/HPF in endometrial stroma affect pregnancy outcome of infertile women?

PROBLEM: The definition of chronic endometritis (CE) differs among studies, and currently, there is no accepted consensus. This study aimed to establish the minimum number of immunohistochemical analysis of CD138+ plasma cells to identify a clinically relevant CE. METHOD OF STUDY: We performed a retrospective study on 716 infertile patients who never did CE analysis and respective antibiotic treatment before. Samples were obtained by endometrial scratching in the mid-luteal phase before IVF-ET treatment. The number and distribution of CD138+ cells were analyzed by immunohistochemistry. Thirty high-power fields (HPF) were evaluated for each sample. Patients were classified in 2 main groups: (a) CD138low (<5 CD138+ cells in all HPFs), (b) CD138high (≥5 CD138+ cells in at least one HPF). Pregnancy outcome was compared among the groups. RESULTS: In the CD138high group, ß-hCG positive rate, clinical pregnancy rate and live birth rate were significantly decreased (P = .04, P = .01, P = .04, respectively). Also after adjusting for patient age, body mass index (BMI), and clinical characteristics, the ß-hCG positive rate (P = .05), clinical pregnancy rate (P = .01) and live birth rate (P = .02) were significantly lower in the CD138high than those in the CD138low group. Within the CD138low group, these parameters were not significantly different between patients without any plasma cells and patients with up to 4 plasma cells/HPF. CONCLUSION: We conclude that immunohistochemical analysis of CD138+ cells is a reliable method to detect CE which can be identified by the presence of ≥5 plasma cells in at least one out of 30 HPF.

Pregnancy Outcomes of Women With Polycystic Ovary Syndrome for the First In Vitro Fertilization Treatment: A Retrospective Cohort Study With 7678 Patients.

Background: The risk of adverse pregnancy outcomes is increased by having a polycystic ovary syndrome (PCOS) diagnosis. However, the confounders in previous studies preclude firm conclusions, and further studies are warranted. Objectives: To investigate whether PCOS affects pregnancy outcomes and complications in infertile women undergoing their first in vitro fertilization (IVF) treatment, taking into account important confounders. Methods: We performed a retrospective cohort study of 7,678 infertile women, including 666 women with PCOS and 7,012 controls undergoing their first IVF treatment at a private fertility center from January 2010 to December 2017. Our main outcome was the impact of PCOS on adverse pregnancy outcomes (miscarriage, preterm delivery, pregnancy-induced hypertension) and pregnancy outcomes (live birth rate, clinical pregnancy rate, implantation rate). PCOS effects were summarized by adjusted odds ratios (aORs) with 95% confidence intervals (CIs) after controlling for maternal characteristics. Results: After adjusting for differences in maternal age, BMI, infertility duration, total dose of gonadotropin, serum E2 and endometrial thickness on the day of hCG trigger, number of fertilized occytes, number of embryos transferred, embryo type (cleavage-stage embryo or blastocyst) and quality, women with PCOS had an increased risk of developing unfavorable pregnancy complications, including miscarriage (aOR 1.629, 95% CI 1.240-2.141), very preterm delivery (< 32 weeks) (aOR 2.072, 95% CI 1.133-3.791). For pregnancy outcomes, PCOS was associated with higher clinical pregnancy rate (aOR 1.248, 95% CI 1.038-1.501) and implantation rate (aOR 1.238, 95% CI 1.030-1.489) after adjusting for the above-mentioned confounders. Conclusions: Women with PCOS are at increased risk of adverse pregnancy outcomes after adjusting for differences in maternal characteristics. These women may need more frequent medical consultants and management during pregnancy and parturition.

Abnormal Expression of Indoleamine 2, 3-Dioxygenase in Human Recurrent Miscarriage.

Indoleamine 2, 3-dioxygenase (IDO), an immunosuppressive enzyme that mediates the conversion of tryptophan to kynurenine, was shown to play a key role in placental development during normal pregnancy. However, little is known about the pattern of IDO expression in the endometrium and its attendant functional significance in pregnancies complicated with recurrent miscarriage (RM). Immunohistochemical studies of IDO, Foxp3, CD56, and CD163 expression were performed in endometrial samples from women with RM and healthy fertile controls. Our study found that IDO was localized in glandular epithelial cells, surface epithelial cells, and a small number of cells within the stromal compartment (including stromal cells and leukocytes) in endometrium. Indoleamine 2, 3-dioxygenase expression in the RM group was significantly lower than control group. The Foxp3 and CD56 expression were significantly increased with the elevated IDO expression in controls but not in RM. The percentage of Foxp3 + Tregs was significantly correlated with the level of IDO expression in the control group. Comparatively, no correlation was found between the percentage of CD56 + cells, CD163 + cells, and the level of IDO expression, no matter in controls and RM patients. This study demonstrated that the downregulation of IDO expression and noncoordinated association between IDO and other endometrial immune cells were associated with RM. Our findings provide insights into the contribution of IDO in immune regulation to maintain normal pregnancy, which could be used to develop potential therapeutic methods for RM.