The impact of previous conservative treatment of atypical hyperplasia on pregnancy outcomes after IVF/ICSI-embryo transfer: a propensity score-matched retrospective cohort study.

STUDY QUESTION: Do women have worse pregnancy and neonatal outcomes of IVF/ICSI-fresh embryo transfer (ET) after conservative treatment of atypical hyperplasia (AH)? SUMMARY ANSWER: AH has no impact on live birth but is associated with increased risks of pregnancy loss and preterm delivery (PTD). WHAT IS KNOWN ALREADY: AH is a precancerous lesion of endometrial cancer. Several recognized AH risk factors include nulliparity, increased body mass index, ovulation disorders, diabetes mellitus, and others. As such, patients are suggested to attempt conception upon achieving AH regression. Recently, successful pregnancies with IVF/ICSI have been increasingly reported. STUDY DESIGN, SIZE, DURATION: Forty-two patients with AH regression and 18 700 women with no evidence of endometrial abnormality, who underwent their first autologous oocytes' retrieval and fresh ET cycles of IVF/ICSI in the Center for Reproductive Medicine, Shandong University, from May 2008 to July 2021, were retrospectively enrolled. PARTICIPANTS/MATERIALS, SETTING, METHODS: First, 42 AH patients were propensity score matched with control women (n = 168) at a 1:4 ratio. Reproductive outcomes and maternal/neonatal complications were compared between the matched pairs. Binary logistic regression analyses were conducted to assess odds ratios (ORs) of AH for live birth, pregnancy loss, and PTD from AH women and all 18 700 eligible controls. MAIN RESULT AND THE ROLE OF CHANCE: Patients with AH achieved a numerically lower live birth rate (LBR) as compared to the matched controls, but without significant difference (26% versus 37%, P = 0.192). However, compared with the matched controls, AH patients showed significantly higher rates of pregnancy loss (52% versus 21%, P = 0.003) and PTD (45% versus 16%, P = 0.041). Further analyses revealed a statistically significantly increased rate of late pregnancy loss (17% versus 3%, P = 0.023), but not early miscarriage (35% versus 18%, P = 0.086), in the AH group. Furthermore, after correcting for potential confounders, the likelihood of a live birth in AH patients narrowly failed to be statistically significantly different from controls (adjusted OR [aOR]: 0.51, 95% CI: 0.25-1.04, P = 0.064). Nonetheless, the logistic regression reconfirmed that AH was an independent risk factor for pregnancy loss (aOR: 3.62, 95% CI: 1.55-8.46, P = 0.003), late pregnancy loss (aOR: 9.33, 95% CI: 3.00-29.02, P < 0.001), and PTD (aOR: 5.70, 95% CI: 1.45-22.38, P = 0.013). LIMITATIONS, REASONS FOR CAUTION: Selection bias was an inherent drawback of this study. First, because of the low AH prevalence among women receiving IVF/ICSI treatment, and consequently, limited sample size, the relationship between AH with LBR and adverse complications might be concealed and underestimated. Hence, the results should be interpreted cautiously. Similarly, the impacts of diverse clinical features of AH patients on the pregnancy outcomes need further studies in a larger population. Second, although most data used in this study were obtained by reviewing the medical records, missing data did exist and so did the recall bias. Third, although the propensity score matching and multivariable logistic models were performed collectively in order to minimize potential confounders between AH and controls, the intrinsic disadvantages of the retrospective nature of this study could not be avoided completely, and additional confirmation bias might be induced with reduplication of statistical analyses. WIDER IMPLICATION OF THE FINDINGS: Our results highlight the necessity of adequate counseling and intensive pregnancy monitoring for AH individuals and their families. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by grants from the National Key Research & Developmental Program of China (2022YFC2703800), the Natural Science Foundation of Shandong Province (ZR2022MH009), and Projects of Medical and Health Technology Development Program in Shandong Province (202005010520, 202005010523). There are no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Conformally Large-Area Single-Crystal Piezocomposites with High Performance for Acoustic Transducers.

Large-area and conformal piezoelectric elements are highly desired for acoustic transducers to possess a large power source level and wide detecting range. To date, single-crystal piezocomposites attract much attention on enhancing the power source level and bandwidth for next-generation acoustic transducers, owing to their higher piezoelectric and electromechanical coupling properties compared to traditional piezocomposites. Unfortunately, it is still challenging to achieve large-area and conformal single-crystal piezocomposites because of the fragile nature, large anisotropy, and the limited grown size of piezoelectric single crystals. Here, we successfully fabricate the conformally large-area single-crystal piezocomposite with an area of 160 × 50 mm2 and a bending angle of 162° by a modified 3D-printing-assisted inserting method. The single-crystal piezocomposite exhibits a high thickness electromechanical coupling factor kt of 85% and a large piezoelectric coefficient d33 of 1150 pC/N, surpassing those of the reported large-area piezocomposites. The influence of the volume fraction and curvature radius of single-crystal PCs and acoustic transducers was investigated. Furthermore, we designed an acoustic transducer based on the conformal single-crystal piezocomposite. Benefiting from the excellent piezoelectric and electromechanical properties of the single-crystal piezocomposite, the transducer indicates a high maximum transmitting voltage response of 171.8 dB. Especially, its bandwidth (-3 dB) achieves 60 kHz with a resonant frequency of 292 kHz, which is about 1.8 times superior to the conformal acoustic transducer based on the ceramic piezocomposite with a similar resonant frequency. This work may benefit the future design and fabrication of high-performance and complex-shape piezoelectric composites as key materials for next-generation transducers.

Relationship Between Plasma Aldosterone Concentrations and Non-Alcoholic Fatty Liver Disease Diagnosis in Patients with Hypertension: A Retrospective Cohort Study.

Objective: To investigate the association between plasma aldosterone concentration (PAC) and non-alcoholic fatty liver disease (NAFLD) diagnosis in Chinese hypertensive patients. Methods: We conducted a retrospective study of all patients diagnosed with hypertension between January 1, 2010, and December 31, 2021. We included 3713 hypertensive patients based on the criteria for inclusion and exclusion. PAC measurement was performed using a radioimmunoassay. NAFLD was diagnosed using abdominal ultrasonography. Cox regression analysis was used to estimate the hazard ratios (HRs) and 95% confidence intervals (CIs) for univariable and multivariable models. A generalized additive model was used to identify nonlinear relationships between PAC and NAFLD diagnosis. Results: A total of 3713 participants were included in the analysis. Over a median follow-up of 30 months, 1572 hypertensive individuals developed new-onset NAFLD. When PAC was used as a continuous variable, the risk of NAFLD increased by 1.04 and 1.24-fold for each 1 ng/dL and 5 ng/dL increase in PAC, respectively. When PAC was considered a categorical variable, the HR for tertile 3 was 1.71 (95% CI, 1.47-1.98, P < 0.001) compared to tertile 1. Overall, there was a J-shaped relationship between PAC and new-onset NAFLD. By fitting a two-piecewise linear regression model and using a recursive algorithm, we identified a PAC inflection point at 13 ng/dL (log-likelihood ratio test, P = 0.005). In adjusted model 3, for PAC ≥ 13 ng/dL, a 5 ng/dL increase in PAC was associated with a 30% increase in the risk of new-onset NAFLD (95% CI, 1.25-1.35, P < 0.001). Conclusion: The study revealed a non-linear relationship between elevated PAC levels and the incidence of NAFLD in hypertensive patients. Notably, the risk of new-onset NAFLD was significantly increased when PAC levels were ≥13 ng/dL. Larger, prospective studies are necessary to confirm these findings.

Association between the sarcopenia index and the risk of stroke in elderly patients with hypertension: a cohort study.

The purpose of this study was to investigate the relationship between the sarcopenia index (SI) and stroke risk in elderly patients with hypertension. This study included 5145 stroke-free elderly hypertensive patients. We used Cox regression models to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) of incident stroke. Over a median follow-up of 38 months, we identified 607 (11.80%) individuals with total stroke, of whom 507 (9.85%) had ischemic stroke and 93 (1.81%) had hemorrhagic stroke. The risk of developing stroke decreased with each quartile of SI; after adjustment for multiple confounders, the HRs for the Q4 group versus the Q1 group were 0.46 (95% CI, 0.35-0.59) for total stroke, 0.46 (95% CI, 0.35-0.61) for ischemic stroke, and 0.33 (95% CI, 0.17-0.64) for hemorrhagic stroke. Restricted cubic spline analysis also demonstrated a cumulative increase in the risk of total stroke with decreases in the SI. The addition of SI to the conventional model for total stroke improved (ΔC-statistics = 0.02), an integrated discrimination improvement of 0.03 (95% CI, 0.02-0.04), and a net reclassification improvement of 0.17 (95% CI, 0.10-0.23). Similar results were observed for ischemic stroke and hemorrhagic stroke. This study found that elevated SI was negatively associated with the risk of stroke in elderly patients with hypertension. Uncovering the causality behind the relationship requires further prospective study.

The effects of unexpected follicular growth and ovulation in artificial cycles: a retrospective cohort study of frozen, single-blastocyst transfer.

OBJECTIVE: To study the effects of unexpected follicular development and ovulation in artificial cycles (ACs) on pregnancy outcomes. DESIGN: A retrospective cohort study. SETTING: A university-affiliated fertility center. PATIENT(S): A total of 1,427 patients who underwent a single, frozen-thawed blastocyst transfer with AC regimens from January 2014 to December 2020 at a university-affiliated fertility center were included. INTERVENTION(S): Unexpected follicular development and ovulation in ACs. MAIN OUTCOME MEASURE(S): Live birth rate (LBR), biochemical pregnancy rate, clinical pregnancy rate, and ongoing pregnancy rate. RESULT(S): A total of 161 patients with unexpected follicular development and ovulation in ACs (ovulation group) and 1,266 patients without growing follicles in ACs (control group) were enrolled. The patients in the ovulation group were older and had higher levels of serum follicle-stimulating hormone and lower levels of serum antimüllerian hormone. After propensity score matching, the baseline characteristics between the 2 groups were comparable and no significant difference was observed in the LBR (ovulation group, 39.0% vs. control group, 39.0%), biochemical pregnancy rate (ovulation group, 60.3% vs. control group, 58.2%), clinical pregnancy rate (ovulation group, 53.4% vs. control group, 50.7%), or ongoing pregnancy rate (ovulation group, 42.5% vs. control group, 40.4%). Moreover, the patients in the ovulation group showed a lower risk of hypertensive disorders of pregnancy (HDP) (1.6% vs. 15.3%). A subgroup analysis of women who delivered singleton live-born babies also demonstrated that unexpected follicular development and ovulation in ACs was associated with a decreased risk of HDP (adjusted odds ratio, 0.070; 95% confidence interval, 0.007-0.712) and an increased risk of large-for-gestational-age infants (adjusted odds ratio, 4.046; 95% confidence interval, 1.319-12.414). CONCLUSION(S): Women with unexpected follicular development and ovulation during single frozen-thawed blastocyst transfer with AC regimens had a similar LBR and a reduced risk of HDP compared with those with routine AC regimens, and singleton neonates had an increased risk of being large for gestational age.

Association between the geriatric nutritional risk index and the risk of stroke in elderly patients with hypertension: A longitudinal and cohort study.

Objective: We aimed to investigate the association between the GNRI and the risk of stroke in elderly patients with hypertension. Methods: A total of 5312 elderly hypertensive patients free of history of stroke were included. Multivariate Cox models were used to calculate hazard ratios (HRs) and their 95% confidence intervals (CIs) for stroke and its subtypes. Results: The average time of follow-up was 3.8 years, and the median time was 3.2 years. We identified 640 individuals with stroke, of whom 526 had an ischemic stroke (IS) and 114 had a hemorrhagic stroke (HS). After adjusting for confounding variables, compared with participants in the lowest quartile of the GNRI, those in the third and fourth quartiles were associated with a decreased risk of stroke (adjusted HR 0.72, 95% CI 0.58-0.90, and adjusted HR 0.58, 95% CI 0.46-0.74, respectively, P for trend < 0.001). Similar results were found for IS and HS. Moreover, there were L-shaped associations of GNRI with new-onset HS (P for non-linearity = 0.034). Multiple sensitivity analyses and stratified analyses did not materially change the results. Conclusions: In summary, we found that a lower GNRI was associated with a higher risk of incident stroke in elderly hypertensive patients. Additional prospective data collection is required to confirm our findings.

Relationship of the metabolic score for insulin resistance and the risk of stroke in patients with hypertension: A cohort study.

Background: The current status of the dose-response relationship between the metabolic score for insulin resistance (METS-IR) and new-onset stroke in hypertensive patients and its subtypes is unclear. This study aimed to determine the association between METS-IR and incident stroke and its subtypes within a cohort of Chinese hypertensive patients. Methods: A total of 14032 hospitalized patients with hypertension from January 1, 2010, to December 31, 2021, were included in this retrospective cohort study. Cox models and restricted cubic splines were applied to determine the association between METS-IR and the risk of stroke. Results: During a median follow-up of 4.80 years, 1067 incident stroke cases occurred. Patients in the highest quartile group of METS-IR levels exhibited a higher risk of stroke (HR, 1.80; 95% CI, 1.50-2.17) and ischemic stroke (HR, 1.96; 95% CI, 1.60-2.42) than those in the lowest quartile group. However, no significant associations were observed between METS-IR and the risk of hemorrhagic stroke. Restricted cubic spline analysis suggested a nearly J-shaped association between METS-IR and risk of stroke and ischemic stroke (P for nonlinearity < 0.001). METS-IR did produce a significant improvement in the C statistic when added to the basic model (from 0.637 to 0.664, P < 0.001). Notably, the addition of METS-IR to the basic model resulted in a significant improvement in predicting incident total stroke and ischemic stroke. Conclusions: This cohort study suggests a relationship between METS-IR and the risk of stroke and ischemic stroke. Further studies are required to elucidate the underlying mechanisms.

The Use of Aspirin Increases the Risk of Major Adverse Cardiac and Cerebrovascular Events in Hypertensive Patients with Obstructive Sleep Apnea for the Primary Prevention of Cardiovascular Disease: A Real-World Cohort Study.

(1) Background: Hypertensive patients with obstructive sleep apnea (OSA) are at high risk for cardiovascular diseases (CVDs), and the utility of aspirin for primary cardiovascular prevention in this population remains uncertain. (2) Methods: In this retrospective cohort study using data from the Urumchi Hypertension Database (UHDATA), hypertensive patients older than 18 years old with a first-time diagnosis of OSA were divided into three groups depending on aspirin history. Major adverse cardiac and cerebrovascular events (MACCE) were the primary outcome. Secondary outcomes included MACCE components, ischemic events, cardiac events, cerebrovascular events, and gastrointestinal bleeding risk. The inverse probability of treatment weighting (IPTW) method was used to balance the confounding factors among the groups, and the Cox proportional hazards model was used to calculate the hazard ratio (HR) and 95% confidence interval (CI). (3) Results: In persistent aspirin users, the risk of MACCE events (HR 2.11, 95%CI 1.23-3.63), ischemic events (HR 2.58, 95%CI 1.42-4.69), cerebrovascular events (HR 2.55, 95%CI 1.44-4.51), and non-fatal cerebral infarction (HR 3.14, 95%CI 1.69-5.84) was significantly elevated. (4) Conclusions: Continuous aspirin use increases the incidence of cardiovascular adverse events in hypertensive patients with OSA receiving aspirin for primary prevention of cardiovascular disease.

Association between first-trimester subchorionic hematoma detected at 6-8 weeks of gestation and pregnancy outcomes after fresh embryo transfers: a propensity score-matching cohort study.

PURPOSE: This study aimed to investigate the association between first-trimester subchorionic hematoma (SCH) detected at 6-8 weeks of gestation after fresh embryo transfers and adverse pregnancy outcomes. METHODS: We performed a retrospective cohort involving 3074 patients. All of them acquired singleton pregnancies after fresh embryo transfers in the first in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. According to first-trimester ultrasound examinations at 6-8 weeks of gestation, we divided patients into SCH and non-SCH groups and compared their perinatal outcomes. Symptomatic patients with vaginal bleeding and asymptomatic patients were analyzed separately, and propensity score matching (PSM) and multivariable regression were adopted to control potential confounding factors. RESULTS: The incidence of SCH was 17.1% in 3074 women, and vaginal bleeding occurred in 92 SCH patients and 215 control patients. In the asymptomatic cohort, 415 women with SCH and 807 women without SCH were finally included after PSM. No significant differences were observed in livebirth rate (91.3% vs 92.9%, P = 0.314), miscarriage rate (8.4% vs 6.7%, P = 0.267), and preterm birth rate (4.8% vs 5.7%, P = 0.519) between two groups. Secondary outcomes including gestational hypertension or preeclampsia, gestational diabetes mellitus (GDM), gestational age (GA) at delivery, mode of delivery, sex of newborns and birthweight of newborns were comparable. For symptomatic patients, both univariable and multivariable regression analysis showed no significant association between SCH and pregnancy outcomes. A subgroup analysis including patients with SCH illustrated the symptom of vaginal bleeding rather than hematoma size was associated with livebirth. CONCLUSION: First-trimester SCH detected at 6-8 weeks of gestation was not associated with adverse pregnancy outcomes in singleton pregnancies after fresh embryo transfers. Vaginal bleeding was the risk factor of pregnancy loss for patients with SCH.

Hepatic Steatosis Index and the Risk of Type 2 Diabetes Mellitus in China: Insights from a General Population-Based Cohort Study.

Purpose: In the Chinese population, we looked at the relationship between the hepatic steatosis index (HSI) and the risk of type 2 diabetes mellitus (T2DM). Methods: To evaluate the association between HSI and the risk of T2DM, Cox regression models were employed. Hazard ratios (HR) and 95 percent confidence intervals (CI) were computed. A stratified analysis with interaction testing was also carried out. Additionally, we evaluated the incremental predictive value of the HSI over the established risk factors using the C-statistic, the IDI, and the NRI. Results: During a median follow-up period of 2.97 years, 433 (1.97%) participants developed new-onset T2DM. The smoothing curve fit plot showed a positive correlation between HSI and the risk of T2DM. After adjusting for all noncollinear variables, the risk of T2DM increased by 62% for every 1 standard deviation (SD) increase in HSI. Subgroup analysis indicated that higher HSI levels were associated with a higher risk of T2DM in those aged < 40 years. The addition of HSI enhanced the reclassification and discrimination of established risk factors, with an IDI of 0.027 and an NRI of 0.348 (both P < 0.001). Conclusion: Our findings suggest that an elevated HSI is substantially associated with a greater risk of T2DM in the Chinese population. HSI has the potential to be an available and supplementary monitoring method for the management of T2DM risk stratification in the Chinese population.

Associations of the Cardiometabolic Index with the Risk of Cardiovascular Disease in Patients with Hypertension and Obstructive Sleep Apnea: Results of a Longitudinal Cohort Study.

Purpose: We aimed to explore the relationship between the cardiometabolic index (CMI) and cardiovascular disease (CVD) and its subtypes (coronary artery disease and stroke) in patients with hypertension and obstructive sleep apnea (OSA). Methods: We conducted a retrospective cohort study enrolling 2067 participants from the Urumqi Research on Sleep Apnea and Hypertension study. The CMI was calculated as triglyceride to high-density lipoprotein cholesterol ratio × waist-to-height ratio. Participants were divided into three groups (T1, T2, and T3) according to the tertile of CMI. The Kaplan-Meier method helped to calculate the cumulative incidence of CVD in different groups. We assessed the association of CMI with the risk of CVD and CVD subtypes by estimating hazard ratios (HRs) and 95% confidence intervals (CIs) using Cox models. Results: During a median follow-up of 6.83 years (interquartile range: 5.92-8.00 years), 326 incident CVD were identified, including 121 incident stroke and 205 incident coronary heart disease (CHD). Overall, after adjusting for confounding variables, CMI was positively associated with the risk of new-onset CVD (per SD increment, adjusted HR: 1.31; 95% CI: 1.20, 1.43), new-onset CHD (per SD increment, adjusted HR: 1.33; 95% CI: 1.20, 1.48), and new-onset stroke (per SD increment, adjusted HR: 1.27; 95% CI: 1.10, 1.47). Similar results were obtained in various subgroup and sensitivity analyses. Adding CMI to the baseline risk model for CVD improved the C-index (P < 0.001), continuous net reclassification improvement (P < 0.001), and integrated discrimination index (P < 0.001). Similar results were observed for CHD and stroke. Conclusion: There was a positive association between CMI levels and the risk of new-onset CVD in patients with hypertension and OSA. This finding suggests that CMI may help identify people at high risk of developing CVD.

Autophagy regulates differentiation of ovarian granulosa cells through degradation of WT1.

Ovarian granulosa cells (GCs) proliferate and differentiate along with follicular growth, and this is indispensable for oocyte development and female fertility. Although the role of macroautophagy/autophagy in ovarian function has been reported, its contribution to the regulation of GC characteristics remains elusive. The siRNA-mediated knockdown of two key autophagy-related genes ATG5 and BECN1 and the autophagy inhibitor chloroquine were used to interfere with autophagy in GCs. Inhibition of autophagy both genetically and pharmacologically resulted in decreased expression of genes associated with GC differentiation, including CYP19A1/Aromatase and FSHR, as well as in reduced estradiol synthesis. Mechanistically, when autophagy was disrupted, the transcription factor WT1 accumulated in GCs due to its insufficient degradation by the autophagic pathway, and this inhibited GC differentiation. Finally, decreased expression of several autophagy-related genes, as well as reduced LC3-II:LC3-I and elevated SQSTM1/p62 protein levels, which are indications of decreased autophagy, were detected in GCs from biochemical premature ovarian insufficiency patients. In summary, our study reveals that autophagy regulates the differentiation of ovarian GCs by degrading WT1 and that insufficient autophagy might be involved in ovarian dysfunction.Abbreviations: ATG: autophagy related; bPOI: biochemical premature ovarian insufficiency; CHX: cycloheximide; Co-IP: co-immunoprecipitation; CQ: chloroquine; E2: estradiol; FSH: follicle stimulating hormone; FSHR: follicle stimulating hormone receptor; GC: granulosa cell; LIR: LC3-interacting region; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; POI: premature ovarian insufficiency; RAP: rapamycin; siRNA: small interfering RNA; WT1: WT1 transcription factor.

lncRNA DDGC participates in premature ovarian insufficiency through regulating RAD51 and WT1.

The list of long non-coding RNAs (lncRNAs) that participate in the function of ovarian granulosa cells (GCs) is rapidly expanding, but the mechanisms through which lncRNAs regulate GC function are not yet fully understood. Here, we recognized a minimally expressed lncRNA RP4-545C24.1 (which we named DDGC) in GCs from patients with biochemical premature ovarian insufficiency (bPOI). We further explored the role of lncRNA DDGC in GC function and its contribution to the development of bPOI. Mechanistically, silencing DDGC downregulated RAD51 by competitively binding with miR-589-5p, and this resulted in significant inhibition of DNA damage repair capacity. In addition, decreased expression of DDGC promoted ubiquitin-mediated degradation of Wilms tumor 1 (WT1) protein through interactions with heat shock protein 90 (HSP90), which led to aberrant differentiation of GCs. Moreover, DDGC was able to ameliorate the etoposide-induced DNA damage and apoptosis in vivo. Taken together, these findings provide new insights into the contribution of lncRNAs in POI pathogenesis.

Preimplantation genetic testing is not a preferred recommendation for patients with X chromosome abnormalities.

STUDY QUESTION: Should women with X chromosome abnormalities (XCAs) be recommended to have embryos selected by both morphological and cytogenetic assessment through preimplantation genetic testing (PGT) rather than morphological assessment only in conventional IVF/ICSI treatment? SUMMARY ANSWER: PGT is not a preferred recommendation for women with XCAs in the absence of other PGT indications. WHAT IS KNOWN ALREADY: XCAs are the most frequent sort of chromosomal aberrations in infertile women. Patients with a complete or partial absence of one X chromosome, diagnosed as Turner Syndrome (TS), demonstrate low spontaneous pregnancy rates (5-7%) and high miscarriage rates (22.8-30.8%), as well as high chances of birth defects (20%). PGT is known to improve pregnancy rates and decrease the incidence of miscarriage in couples with chromosomal aberrations such as Robertsonian and reciprocal translocations and Klinefelter Syndrome. STUDY DESIGN, SIZE, DURATION: A retrospective cohort study was conducted with 394 women with XCAs and undergoing their first oocyte retrieval and first embryo transfer cycle from June 2011 to August 2019 in the Reproductive Hospital Affiliated to Shandong University. PARTICIPANTS/MATERIALS, SETTING, METHODS: Pregnancy outcomes were compared between the conventional IVF/ICSI group (n = 284) and the PGT group (n = 110) in the first fresh or frozen embryo transfer cycle for each woman with XCAs. Three platforms were applied in PGT: fluorescence in situ hybridisation (FISH, n = 34), array comparative genomic hybridisation (aCGH, n = 24) and next-generation sequencing (NGS, n = 51). The embryo aneuploidy rate and distribution of embryonic chromosomal aberrations revealed by aCGH or NGS were analysed and stratified by maternal age and type of XCAs to assess the effect of maternal XCAs on embryo karyotypes. MAIN RESULT AND THE ROLE OF CHANCE: The live birth rate (LBR) per embryo transfer was similar between the PGT group and IVF/ICSI group both in the first cycle of fresh or frozen embryo transfer respectively (39.13% in PGTFISH vs 42.58% in IVF/ICSI, Padj=0.558; 66.67% in PGTFISH vs 52.08% in PGTaCGH/NGS vs 53.06% in IVF/ICSI, Padj=0.756), as was the clinical pregnancy rate (60.87% in PGTFISH vs 50.97% in IVF/ICSI, Padj =0.672; 88.89% in PGTFISH vs 58.33% in PGTaCGH/NGS vs 69.39% in IVF/ICSI, Padj =0.480) and the pregnancy loss rate (35.71% in PGTFISH vs 16.46% in IVF/ICSI, Padj =0.136; 12.50% in PGTFISH vs 10.71% in PGTaCGH/NGS vs 23.53% in IVF/ICSI, Padj =0.352). The rates of maternal and neonatal complications were also comparable between the PGT and IVF/ICSI groups with fresh and frozen transfers respectively (10.00% vs 8.85%, P = 1.000; 21.74% vs 14.55%, P = 0.272). Intriguingly, the distribution of embryonic chromosome abnormalities was more frequent on autosomes 22 (20.39%), 21 (18.45%) and 16 (17.47%), compared with the X chromosome (8.73%). LIMITATIONS, REASONS FOR CAUTION: Selection bias is an inherent drawback of a retrospective study. First, our participants hosted 4.84% X chromosome mosaicism with few typical somatic anomalies of TS. Second, the incidences of history of recurrent miscarriage and abnormal offspring in the PGT group were higher than in IVF/ICSI group although binary logistic regression analysis was performed to attenuate the modifying effect of confounding factors. Third, FISH performed in this study only used X/Y probes and lacked the reference of autosome, which might have resulted in misdiagnosis and bias. Finally, intrinsic disadvantages could not be totally avoided due to the retrospective nature of this study. WIDER IMPLICATION OF THE FINDINGS: In the current study, comparable pregnancy outcomes were revealed among a large cohort of women with XCAs undergoing their first cycles of PGT or conventional IVF/ICSI treatment. Moreover, the X chromosome abnormality was illustrated to cause no higher frequency of aberrations in embryos. Our data provided perspectives for genetic and reproductive counselling to XCAs individuals and their families. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by National Research and Development Plan (2016YFC1000604 and 2017YFC1001100), the National Natural Science Foundation of China (81701406), Shandong Science Fund for Distinguished Young Scholars (JQ201720), Taishan Scholars Program for Young Experts of Shandong Province (tsqn20161069) and Projects of Medical and Health Technology Development Program in Shandong Province (202005010520, 202005010523 and 2016WS0368). There is no conflict of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Treg deficiency-mediated TH 1 response causes human premature ovarian insufficiency through apoptosis and steroidogenesis dysfunction of granulosa cells.

Immune dysregulation has long been proposed as a component of premature ovarian insufficiency (POI), but the underlying mediators and mechanisms remain largely unknown. Here we showed that patients with POI had augmented T helper 1 (TH 1) responses and regulatory T (Treg ) cell deficiency in both the periphery and the ovary compared to the control women. The increased ratio of TH 1:Treg cells was strongly correlated with the severity of POI. In mouse models of POI, the increased infiltration of TH 1 cells in the ovary resulted in follicle atresia and ovarian insufficiency, which could be prevented and reversed by Treg cells. Importantly, interferon (IFN) -γ and tumor necrosis factor (TNF) -α cooperatively promoted the apoptosis of granulosa cells and suppressed their steroidogenesis by modulating CTGF and CYP19A1. We have thus revealed a previously unrecognized Treg cell deficiency-mediated TH 1 response in the pathogenesis of POI, which should have implications for therapeutic interventions in patients with POI.

LncRNA ZNF674-AS1 regulates granulosa cell glycolysis and proliferation by interacting with ALDOA.

Granulosa cell (GC) is a critical somatic component of ovarian follicles to support oocyte development, while the regulatory role of long noncoding RNA (lncRNA) in GCs is largely unknown. Here, we identified a down-regulated lncRNA ZNF674-AS1 in GCs from patients with biochemical premature ovarian insufficiency (bPOI), and its expression correlates with serum levels of clinical ovarian reserve indicators. Functional experiments showed that ZNF674-AS1 is induced by energy stress, and regulates the proliferation and glycolysis of GCs, which possibly leads to follicular dysfunction. Mechanistically, low-expressed ZNF674-AS1 reduced the enzymatic activity of aldolase A (ALDOA), concomitant with promoting the association between ALDOA and v-ATPase to activate the lysosome localized AMP-activated protein kinase (AMPK). These findings identified a new lncRNA-ALDOA complex through which ZNF674-AS1 exerts its functions, expanding the understanding of epigenetic regulation of GCs function and POI pathogenesis.

lncRNA GCAT1 is involved in premature ovarian insufficiency by regulating p27 translation in GCs via competitive binding to PTBP1.

Dysfunction of granulosa cells (GCs) leading to follicle atresia has been extensively studied as a major cause of premature ovarian insufficiency (POI), but the regulatory role of long non-coding RNAs (lncRNAs) in this process is still poorly understood. Here, we show that the lncRNA LINC02690 or GCAT1 (granulosa cell-associated transcript 1) is downregulated in GCs from patients with biochemical POI (bPOI), and we show a significant correlation between downregulated GCAT1 and serum levels of follicle-stimulating hormone and anti-Müllerian hormone. Downregulation of GCAT1 inhibited G1/S cell cycle progression and thus inhibited the proliferation of GCs. Mechanistically, we show that GCAT1 competes with cyclin-dependent kinase inhibitor 1B (CDKN1B) mRNA for polypyrimidine tract-binding protein 1 (PTBP1) binding, and thus decreased GCAT1 might promote PTBP1 binding to CDKN1B mRNA and thereby initiate CDKN1B protein (p27) translation. Together, our results suggest that downregulation of GCAT1 under conditions of bPOI inhibits the proliferation of GCs through PTBP1-dependent p27 regulation, thus suggesting a novel form of lncRNA-mediated epigenetic regulation of GC function that contributes to the pathogenesis of POI.

MiR-148a-3p may contribute to flawed decidualization in recurrent implantation failure by modulating HOXC8.

PURPOSE: To evaluate whether miR-148a-3p overexpression is associated with disrupted decidualization of recurrent implantation failure (RIF). METHODS: Endometrial miRNA and mRNA expression profiles during the implantation window derived from women with and without RIF were identified using microarray and RT-qPCR. Immortalized human endometrial stromal cells (HESCs) were cultured for proliferation and in vitro decidualization assays after enhancing miR-148a-3p expression or inhibiting putative target gene homeobox C8 (HOXC8) expression. RT-qPCR, western blot, and luciferase reporter assays were used to confirm the relationship between miR-148a-3p and HOXC8 gene. RESULTS: MiR-148a-3p was significantly upregulated in RIF endometrial tissues. Forced expression of miR-148a-3p notably attenuated HESC in vitro decidualization. Mechanistic studies revealed that miR-148a-3p directly bounds to the HOXC8 3' untranslated region (3'UTR) and suppressed HOXC8 expressions in both mRNA and protein levels. Further investigations demonstrated that inhibition of HOXC8 in HESCs induced similar effects on decidual process as those induced by miR-148a-3p overexpression. CONCLUSION: Taken together, our findings suggested that elevated miR-148a-3p might account for flawed decidualization in RIF by negatively regulating HOXC8, raising the possibility that miR-148a-3p might be a novel therapeutic target in RIF.

MicroRNA-127-5p impairs function of granulosa cells via HMGB2 gene in premature ovarian insufficiency.

Distinct microRNA (miRNA) profiles have been reported in premature ovarian insufficiency (POI), but their functional relevance in POI is not yet clearly stated. In this study, aberrant expressions of miR-127-5p and high mobility group box 2 (HMGB2) were observed by microarrays in granulosa cells (GCs) from biochemical POI (bPOI) women and further confirmed by a quantitative reverse-transcription polymerase chain reaction. Immortalized human granulosa cell line and mouse primary ovarian GCs were used for functional validation. Orthotopic mouse model was established to examine the role of miR-127-5p in vivo. Finally, the expression of miR-127-5p was measured in the plasma of bPOI women. The receiver operating characteristic curve analysis was performed to determine the indicative role of miR-127-5p for ovarian reserve. Results showed the upregulation of miR-127-5p was identified in GCs from bPOI patients. It inhibited GCs proliferation and impaired DNA damage repair capacity through targeting HMGB2, which was significantly downregulated in GCs from the same cohort of cases. miR-127-5p was confirmed to attenuate DNA repair capability via HMGB2 in mouse ovary in vivo. Intriguingly, the upexpression of miR-127-5p was also detected in plasma of bPOI individuals, suggesting that miR-127-5p could be a promising indicator for bPOI. Taken together, our results discovered the deleterious effects of miR-127-5p on GCs function and its predictive value in POI process. The target gene HMGB2 could be considered as a new candidate for POI. This study highlights the importance of DNA repair capacity for ovarian function and sheds light on the epigenetic mechanism in the pathogenicity of POI.