Publisher Correction: Chromatin analysis in human early development reveals epigenetic transition during ZGA.

In this Letter, the 'Open chromatin' label in Fig. 4a should have been centred above the first three columns, and the black horizontal line underneath the label should have been removed. In addition, there should have been a vertical black line between the last two sets of panels for consistency. Minor changes have also been made to Fig. 1 and to the legend of Fig. 3. These errrors have been corrected online, and see Supplementary Information to the accompanying Amendment for the original Fig. 4.

Chromatin analysis in human early development reveals epigenetic transition during ZGA.

Upon fertilization, drastic chromatin reorganization occurs during preimplantation development 1 . However, the global chromatin landscape and its molecular dynamics in this period remain largely unexplored in humans. Here we investigate chromatin states in human preimplantation development using an improved assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) 2 . We find widespread accessible chromatin regions in early human embryos that overlap extensively with putative cis-regulatory sequences and transposable elements. Integrative analyses show both conservation and divergence in regulatory circuitry between human and mouse early development, and between human pluripotency in vivo and human embryonic stem cells. In addition, we find widespread open chromatin regions before zygotic genome activation (ZGA). The accessible chromatin loci are readily found at CpG-rich promoters. Unexpectedly, many others reside in distal regions that overlap with DNA hypomethylated domains in human oocytes and are enriched for transcription factor-binding sites. A large portion of these regions then become inaccessible after ZGA in a transcription-dependent manner. Notably, such extensive chromatin reorganization during ZGA is conserved in mice and correlates with the reprogramming of the non-canonical histone mark H3K4me3, which is uniquely linked to genome silencing3-5. Taken together, these data not only reveal a conserved principle that underlies the chromatin transition during mammalian ZGA, but also help to advance our understanding of epigenetic reprogramming during human early development and in vitro fertilization.

Decreased NOTCH1 signaling activated autophagy in the mid-secretory endometrium of patients with recurrent implantation failure†.

Recurrent implantation failure severely impairs fertility in females of childbearing age, which poses a great challenge to assisted reproductive technology, and its etiology is still unclear. Several studies have demonstrated that endometrial autophagy takes an important part in human endometrial receptivity, but its role in recurrent implantation failure remains largely unknown. Here, we collected mid-secretory endometrial tissue from recurrent implantation failure patients and fertile controls during menstruation and early pregnancy. Immunohistochemistry, western blotting, and quantitative real-time PCR were performed to compare the expression of microtubule-associated protein 1 light chain 3B, sequestosome 1, NOTCH1 signaling pathway members, and endometrial receptivity markers between recurrent implantation failure and control groups. In addition, to assess endometrial autophagy, transmission electron microscopy was used to observe autophagosomes. By RNA interference, we further investigated the effects of NOTCH1 on autophagy in Ishikawa cells. We found that endometrial autophagy was upregulated in the mid-secretory and decidual phases than in the early-proliferative phase. Compared to the control group, more autophagosomes were observed in the mid-secretory endometrium of recurrent implantation failure patients, which was accompanied by the downregulation of NOTCH1 signaling pathway members and endometrial receptivity markers. Moreover, knockdown of NOTCH1 impaired the receptivity of Ishikawa cells via protein kinase B/mammalian target of rapamycin pathway-mediated autophagy activation. Our data suggested that abnormally elevated autophagy and decreased NOTCH1 signaling pathway activity were observed in the mid-secretory endometrium of patients with recurrent implantation failure, perhaps due to decreased NOTCH1 pathway-mediated autophagy activation in endometrial cells impairing receptivity.

Effective protection: the embryonic development and clinical outcomes of emergency vitrification of 1246 oocytes and Day 0-Day 5 embryos in a natural disaster.

STUDY QUESTION: Can emergency vitrification protect embryos and oocytes during natural disasters or other events that prevent normal practice to achieve satisfactory embryonic development and clinical outcomes at a later time? SUMMARY ANSWER: Emergency vitrification of oocytes and Day 0-Day 5 (D0-D5) embryos during disasters is a safe and effective protective measure. WHAT IS KNOWN ALREADY: When some destructive events such as floods, earthquakes, tsunamis, and other accidents occur, emergency vitrification in embryo laboratories to protect human embryos, oocytes, and sperm is one of the important measures of an IVF emergency plan. However, there are few detailed reports on emergency vitrification in a state of disaster, especially about oocytes and D0 zygotes. Therefore, the effectiveness and safety of emergency vitrification of oocytes and D0-D5 embryos in disaster states are still unclear. STUDY DESIGN, SIZE, DURATION: A retrospective study was made in the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University from January 2018 to November 2022. The record rainstorms in Zhengzhou, China, caused severe flooding, traffic disruptions, and power outages. From 17:30, 20 July 2021 to 17:30, 21 July 2021, 1246 oocytes and D0-D5 embryos of 155 patients were vitrified whilst the laboratory had only an emergency power supply. PARTICIPANTS/MATERIALS, SETTING, METHODS: As of 21 December 2021, 1149 emergency vitrified oocytes and D0-D5 embryos of 124 patients underwent frozen-thawed embryo transfer (FET). They were divided into the following four groups according to the days of embryo culture in vitro: oocyte group, Day 0-Day 1 (D0-D1) group, Day 2-Day 3 (D2-D3) group, and Day 4-Day 5 (D4-D5) group. Control groups for each were selected from fresh cycle patients who underwent IVF/ICSI from January 2018 to October 2021. Control and emergency vitrification patients were matched on criteria that included age, fertilization method, days of embryonic development, and number and grade of transferred embryos. A total of 493 control patients were randomly selected from the eligible patients and matched with the emergency vitrification groups in a ratio of 4:1. The results of assisted reproduction and follow-up of pregnancy were analyzed. The embryonic development, clinical outcomes, and birth outcomes in each group were statistically analyzed. MAIN RESULTS AND THE ROLE OF CHANCE: A significant difference was observed in fertilization rate (81% versus 72%, P = 0.022) between the oocyte group and the control group. Significant differences were also observed in the monozygotic twin pregnancy rate (10% versus 0%, P = 0.038) and ectopic pregnancy rate (5% versus 0%, P = 0.039) between the D0-D1 group and the control group. No significant differences (P > 0.05) were observed between vitrified oocytes/D0-D1 embryos/D2-D3 embryos and the control group on the number of high-quality embryos (3.17 ± 3.00 versus 3.84 ± 3.01, P = 0.346; 5.04 ± 3.66 versus 4.56 ± 2.87, P = 0.346; 4.85 ± 5.36 versus 5.04 ± 4.64, P = 0.839), the number of usable blastocysts (1.22 ± 1.78 versus 1.21 ± 2.03, P = 0.981; 2.16 ± 2.26 versus 1.55 ± 2.08, P = 0.090; 2.82 ± 3.23 versus 2.58 ± 3.32, P = 0.706), clinical pregnancy rate (56% versus 57%, P = 0.915; 55% versus 55%, P = 1.000; 40% versus 50%, P = 0.488), miscarriage rate (30% versus 15%, P = 0.496; 5% versus 11%, P = 0.678; 17% versus 20%, P = 1.000), and live birth rate (39% versus 49%, P = 0.460; 53% versus 50%, P = 0.772; 33% versus 40%, P = 0.635). No significant differences (P > 0.05) were observed between the D4-D5 group and the control group on clinical pregnancy rate (40% versus 55%, P = 0.645), miscarriage rate (0% versus 18%, P = 1.000), and live birth rate (40% versus 45%, P = 1.000). LIMITATIONS, REASONS FOR CAUTION: The retrospective study design is a limitation. The timing and extent of natural disasters are unpredictable, so the sample size of vitrified oocytes, zygotes, and embryos is beyond experimental control. WIDER IMPLICATIONS OF THE FINDINGS: This study is the first study analyzing embryonic development, clinical outcomes, and birth outcomes of large samples of oocytes, D0 zygotes, and D1-D5 embryos after emergency vitrification under the disaster conditions. The results show that emergency vitrification is a safe and effective protective measure applicable to oocytes and D0-D5 embryos. The embryology laboratories need to be equipped with an emergency uninterrupted power supply capable of delivering for 6-8 h at full load. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the National Natural Science Foundation of China (grant 81871206). The authors declare that they have no conflicts of interest. All authors have completed the ICMJE Disclosure form. TRIAL REGISTRATION NUMBER: N/A.

High sperm DNA fragmentation increased embryo aneuploidy rate in patients undergoing preimplantation genetic testing.

RESEARCH QUESTION: Is high sperm DNA fragmentation (SDF) associated with a high embryonic aneuploidy rate in patients undergoing intracytoplasmic sperm injection (ICSI)-preimplantation genetic testing (PGT)? DESIGN: This was a retrospective study of 426 couples with normal karyotypes undergoing ICSI-PGT at the authors' centre from March 2017 to March 2021. SDF was assessed using the sperm chromatin structure assay. The population was divided into low and high SDF groups according to cut-off values found by the receiver operating characteristic (ROC) curve. A 1:1 ratio propensity score matching (PSM) method was used to control for potential confounding factors, and a generalized linear mixed model was established to evaluate the relationship between SDF and the embryonic aneuploidy rate. RESULTS: The ROC curve indicated a threshold of 30%. In total, 132 couples were included after PSM, and the high SDF group (>30%) had significantly higher SDF (40.74% ± 9.78% versus 15.54% ± 7.86%, P < 0.001) and a higher embryo aneuploidy rate (69.36% versus 53.96%, P < 0.001) compared with the low SDF group (≤30%). The two pronuclear fertilization rate, cleavage rate, rate of high-quality embryos at day 3 rate, blastocyst rate, biochemical pregnancy rate, clinical pregnancy rate, miscarriage rate, live birth rate, caesarean section rate, preterm birth rate, singleton rate and low birthweight rate were similar in both groups (P > 0.05). After PSM, SDF > 30% was significantly correlated with an increased embryo aneuploidy rate after adjusting for all confounding variables (adjusted odds ratio 1.70, 95% CI 1.00-2.88, P = 0.049). CONCLUSIONS: SDF > 30% was associated with an increased embryo aneuploidy rate in couples with normal karyotypes undergoing PGT, but did not affect embryonic and clinical outcomes after transfer of euploid embryos.

Comparison of recombinant human FSH biosimilar QL1012 with Gonal-f® for ovarian stimulation: a phase-three trial.

RESEARCH QUESTION: Are QL1012 and Gonal-f® equivalent in women undergoing ovarian stimulation for assisted reproductive technology (ART)? DESIGN: This multicentre, randomized, assessor-blinded, phase-three trial was conducted at 13 centres in China. Eligible patients were infertile women; age 20-39 years; body mass index 18-30 kg/m2; regular menstrual cycles; and indication for ART. After successful pituitary downregulation, patients were randomly assigned (1:1) to receive QL1012 or Gonal-f®, stratified by age (initial dose of 75-150 IU for women younger than 30 years, 150-225 IU for women aged 30-34 years and 225-300 IU for women aged ≥35 years, subcutaneously, once daily). The primary end point was the number of oocytes retrieved. RESULTS: Between October 2018, and June 2019, 341 patients were included in the per-protocol set. The mean numbers of oocytes retrieved were 14.7 ± 7.0 in the QL1012 group (n = 169) and 13.4 ± 6.1 in the Gonal-f® group (n = 172). Adjusted by analysis of covariance model, the least-squares mean difference was 1.3 oocytes (95% CI -0.1 to 2.7; P = 0.0650), within the pre-defined equivalence margins of ±3.0. Similar results were observed in the full analysis set. Additionally, no statistical differences were found in secondary end points except oestradiol concentration (median 3948.0 pg/ml versus 3545.3 pg/ml; P = 0.0015). Ovarian hyperstimulation syndrome (12.4% versus 13.1 %) and other adverse events were similar between the two groups. CONCLUSIONS: Therapeutic equivalence and similar safety profiles were demonstrated between QL1012 and Gonal-f® in women undergoing ovarian stimulation for ART.

Comparisons of different treatment outcomes in IVF/ET patients with hydrosalpinx: a retrospective study.

AIMS: To explore suggestions for clinicians on the most effective treatment for hydrosalpinx undergoing IVF-ET. MATERIALS AND METHODS: We reviewed 936 women with hydrosalpinx and 6715 tubal infertile women without hydrosalpinx who underwent IVF/ICSI between January 2014 and August 2019 in our center. Hydrosalpinx patients received different treatments including laparoscopic surgery (only salpingectomy and proximal tubal occlusion/ligation were included), ultrasonic-guided aspiration and hysteroscopic tubal occlusion. Outcomes were analyzed by One-way ANOVA, Chi-Square test and logistic regression. RESULTS: The live birth rate (LBR) of laparoscopic surgery was significantly higher compared with hydrosalpinx aspiration (48.3% vs 39.6%, p = .024). The cumulative live birth rate (CLBR) of subsequent laparoscopic surgery was significantly higher compared with subsequent hysteroscopic occlusion (65.1% vs 34.1%, p = .001) and no subsequent treatment (65.1% vs 44.9%, p < .005). Subsequent laparoscopic surgery significantly improved the CLBR of hydrosalpinx patients who received ultrasonic-guided aspiration and didn't get clinical pregnancy in fresh cycles (Odds Ratio (OR) =1.875; 95%CI = 1.041-3.378, p = .036). CONCLUSIONS: Laparoscopic surgery leads to significantly higher LBR than ultrasonic-guided aspiration and significantly higher CLBR than hysteroscopic occlusion and no treatment.

Exogenous brassinosteroid alleviates calcium deficiency induced tip-burn by regulating calcium transport in Brassica rapa L. ssp. pekinensis.

Mini Chinese cabbage (Brassica rapa L. ssp. Pekinensis) plays an important role in the supply of summer vegetables on the plateau in western China. In recent years, tip-burn has seriously affected the yield, quality and commodity value of mini Chinese cabbage. Calcium (Ca2+) deficiency is a key inducer of tip-burn. As a new type plant hormone, brassinolide (BR) is involved in regulating a variety of biotic and abiotic stresses. To explore the alleviation role of BR in tip-burn caused by Ca2+ deficiency, a hydroponic experiment was conducted to study the relationship between BR and Ca2+ absorption and transport. The results showed that foliar spraying with 0.5 µM BR significantly reduced tip-burn incidence rate and disease index of mini Chinese cabbage caused by Ca2+ deficiency. Moreover, the dynamic monitoring results of tip-burn incidence rate showed that the value reached the highest on the ninth day after treatment. BR promoted the Ca2+ transport from roots to shoots and from outer leaves to inner leaves by increasing the activities of Ca2+-ATPase and H+-ATPase as well as the total ATP content, which provided power for Ca2+ transport. In addition, exogenous BR upregulated the relative expression levels of BrACA4, BrACA11, BrECA1, BrECA3, BrECA4, BrCAX1, BrCAS and BrCRT2, whereas Ca2+ deficiency induced down-regulation. In conclusion, exogenous BR can alleviate the Ca2+-deficiency induced tip-burn of mini Chinese cabbage by promoting the transport and distribution of Ca2+.

The Influence of External Parameters on the Ripeness of Pumpkins.

Growing pumpkins in controlled environments, such as greenhouses, has become increasingly important due to the potential to optimise yield and quality. However, achieving optimal environmental conditions for pumpkin cultivation requires precise monitoring and control, which can be facilitated by modern sensor technologies. The objective of this study was to determine the optimal placement of sensors to determine the influence of external parameters on the maturity of pumpkins. The greenhouse used in the study consisted of a plastic film for growing pumpkins. Five different sensors labeled from A1 to A5 measured the air temperature, humidity, soil temperature, soil humidity, and illumination at five different locations. We used two methods, error-based sensor placement and entropy-based sensor placement, to evaluate optimisation. We selected A3 sensor locations where the monitored data were close to the reference value, i.e., the average data of all measurement locations and parameters. Using this method, we selected sensor positions to monitor the influence of external parameters on the maturity of pumpkins. These methods enable the determination of optimal sensor locations to represent the entire facility environment and detect areas with significant environmental disparities. Our study provides an accurate measurement of the internal environment of a greenhouse and properly selects the base installation locations of sensors in the pumpkin greenhouse.

Nitric Oxide Induced by Ammonium/Nitrate Ratio Ameliorates Low-Light Stress in Brassica pekinesis: Regulation of Photosynthesis and Root Architecture.

Low-light intensity affects plant growth and development and, finally, causes a decrease in yield and quality. There is a need for improved cropping strategies to solve the problem. We previously demonstrated that moderate ammonium:nitrate ratio (NH4+:NO3-) mitigated the adverse effect caused by low-light stress, although the mechanism behind this alleviation is unclear. The hypothesis that the synthesis of nitric oxide (NO) induced by moderate NH4+:NO3- (10:90) involved in regulating photosynthesis and root architecture of Brassica pekinesis subjected to low-light intensity was proposed. To prove the hypothesis, a number of hydroponic experiments were conducted. The results showed that in plants exposed to low-light intensity, the exogenous donors NO (SNP) and NH4+:NO3- (N, 10:90) treatments significantly increased leaf area, growth range, and root fresh weight compared with nitrate treatment. However, the application of hemoglobin (Hb, NO scavenger), N-nitro-l-arginine methyl ester (L-NAME, NOS inhibitor), and sodium azide (NaN3, NR inhibitor) in N solution remarkably decreased the leaf area, canopy spread, the biomass of shoot and root, the surface area, and volume and tips of the root. The application of N solution and exogenous SNP significantly enhanced Pn (Net photosynthetic rate) and rETR (relative electron transport rates) compared with solo nitrate. While all these effects of N and SNP on photosynthesis, such as Pn, Fv/Fm (maximum quantum yield of PSII), Y(II) (actual photosynthetic efficiency), qP (photochemical quenching), and rETR were reversed when the application of Hb, L-NAME, and NaN3 in N solution. The results also showed that the N and SNP treatments were more conducive to maintaining cell morphology, chloroplast structure, and a higher degree of grana stacking of low-light treated plants. Moreover, the application of N significantly increased the NOS and NR activities, and the NO levels in the leaves and roots of mini Chinese cabbage seedlings treated with N were significantly higher than those in nitrate-treated plants. In conclusion, the results of this study showed that NO synthesis induced by the appropriate ammonia-nitrate ratio (NH4+:NO3- = 10:90) was involved in the regulation of photosynthesis and root structure of Brassica pekinesis under low-light stress, effectively alleviating low-light stress and contributing to the growth of mini Chinese cabbage under low-light stress.

Comprehensive evaluation on effect of planting and breeding waste composts on the yield, nutrient utilization, and soil environment of baby cabbage.

Treatment of the planting and breeding waste is becoming a big issue due to their significant quantities. Composting could be an effective alternative for planting and breeding waste management which could be used as fertilizer. The purpose of this research was to evaluate the effect of planting and breeding waste on baby cabbage growth and soil properties, to establish a suitable agricultural cycle model for semi-arid area in central Gansu Province. The planting and breeding wastes [sheep manure (SM), tail vegetable (TV), cow manure (CM), mushroom residue (MR) and corn straw (CS)] were used as the raw materials in this study, which were designed 8 compost formulas for composting fermentation. With no fertilization (CK1) and local commercial organic fertilizer (CK2) as the control, the comprehensive evaluation of planting and breeding waste composts on the yield of baby cabbage, fertilizer utilization rate, soil physical and chemical properties and microbial diversity were studied to select the best compost formula suitable for the growth of baby cabbage. And the material flow and energy flow analysis of the circulation model established by the formula were carried out. The results showed that the biological yield and economic yield of baby cabbage, absorption and recycling utilization of total phosphorus (TP) and total potassium (TK) reached the maximum under the formula of SM: TV: MR: CS = 6:2:1:1. Compared with CK2, the formula of SM: TV: MR: CS = 6:2:1:1 significantly increased the richness of soil bacteria and beneficial bacteria Proteobacteria, and decreased the relative abundance of harmful bacteria Olpidiomycota. Principal component analysis showed the comprehensive score of SM: TV: MR: CS = 6:2:1:1 was the best organic compost formula suitable for producing high-quality and high-yield baby cabbage and improving soil environment. Therefore, this formula can be used as a reference organic fertilizer formula for field cultivation of baby cabbage.

Comparison of embryo implantation potential between time-lapse incubators and standard incubators: a randomized controlled study.

RESEARCH QUESTION: What are the potential clinical benefits of embryo culture and assessment in a time-lapse incubator compared with a standard incubator using static assessment? DESIGN: This large multicentre, single-blinded, randomized controlled study included 1224 participants randomly assigned (1:1) to the time-lapse or standard incubator group. In all patients one or two embryos were transferred on day 3. The primary outcome was the implantation rate in the first embryo transfer cycle. Secondary outcomes included the cumulative implantation rate, live birth rate in the first embryo transfer cycle and cumulative live birth rate. RESULTS: Among 1224 participants recruited, 1182 underwent embryo transfer. The number of successfully implanted embryos in the first transfer cycle was significantly higher in the time-lapse incubator group (time-lapse group: 52.35%, standard incubator group: 47.11%, P = 0.014). The implantation rate in the first embryo transfer cycle was still significantly higher in the time-lapse group than the standard incubator group after adjusting for age, body mass index, medical centre and embryo status (relative risk 1.11, 95% confidence interval 1.02-1.20, P = 0.020). However, the cumulative implantation rate, live birth rate in the first embryo transfer cycle and cumulative live birth rate were not statistically different between the groups. CONCLUSIONS: The implantation rate in the first embryo transfer cycle was significantly improved in the time-lapse group, but the effect of the time-lapse system on the cumulative implantation rate or cumulative live birth rate was not significant. The embryo assessment method offered by time-lapse systems rather than an undisturbed environment may play an important role in improving the implantation rate in the first embryo transfer cycle. These results are only applicable to young patients.

Placensin is a glucogenic hormone secreted by human placenta.

FBN1 encodes asprosin, a glucogenic hormone, following furin cleavage of the C-terminus of profibrillin 1. Based on evolutionary conservation between FBN1 and FBN2, together with conserved furin cleavage sites, we identified a peptide hormone placensin encoded by FBN2 based on its high expression in trophoblasts of human placenta. In primary and immortalized murine hepatocytes, placensin stimulates cAMP production, protein kinase A (PKA) activity, and glucose secretion, accompanied by increased expression of gluconeogenesis enzymes. In situ perfusion of liver and in vivo injection with placensin also stimulate glucose secretion. Placensin is secreted by immortalized human trophoblastic HTR-8/SVneo cells, whereas placensin treatment stimulates cAMP-PKA signaling in these cells, accompanied by increases in MMP9 transcripts and activities, thereby promoting cell invasion. In pregnant women, levels of serum placensin increase in a stage-dependent manner. During third trimester, serum placensin levels of patients with gestational diabetes mellitus are increased to a bigger extent compared to healthy pregnant women. Thus, placensin represents a placenta-derived hormone, capable of stimulating glucose secretion and trophoblast invasion.

The effect of endometrial scratch on pregnancy outcomes of frozen-thawed embryo transfer: a propensity score-matched study.

AIMS: To study the effect of endometrial scratch (ES) performed on Days 2 to 5 of the menstrual cycle on pregnancy outcomes of frozen-thawed embryo transfer (FET). MATERIALS AND METHODS: We conducted a retrospective study including 5769 women undergoing FET from January 2018 to August 2019. The endometrial scratching group (group ES) consisted of 827 patients and the control group (group non-ES) comprised 4942 patients. A propensity score matching (PSM) method using the nearest neighbor matching at a proportion of 1:1 was performed. Subsequently, we compared the pregnancy outcomes of each group after PSM. RESULTS: After PSM, there were significant differences in implantation (36.1% vs. 47.0%, p = .005), clinical pregnancy (45.4% vs. 58.3%, p = .007), and live birth (33.5% vs. 47.2%, p = .003) rates in matched patients with two previous implantation failures. In the matched patients with at least three previous implantation failures, we also detected increase rates of implantation (31.0% versus 40.1%, p = .028), clinical pregnancy (39.8% versus 57.0%, p = .019), live birth (32.3% versus 47.3%, p = .036). In contrast, no significant difference in pregnancy outcomes was found in women with no previous implantation failure or only one failure. CONCLUSIONS: Endometrial scratch on day 2 to day 5 of the menstrual cycle in women with at least two previous implantation failures who underwent FET increased their implantation, clinical pregnancy, and live birth rates, and did not increase miscarriage or ectopic pregnancy rates compared to the non-ES group.

BR-Mediated Protein S-Nitrosylation Alleviated Low-Temperature Stress in Mini Chinese Cabbage (Brassica rapa ssp. pekinensis).

Brassinosteroids (BRs), a novel plant hormone, are widely involved in plant growth and stress response processes. Nitric oxide (NO), as an important gas signaling molecule, can regulate target protein activity, subcellular localization and function in response to various stresses through post-translational S-nitrosylation modifications. However, the relationship between BR and NO in alleviating low-temperature stress of mini Chinese cabbage remains unclear. The hydroponic experiment combined with the pharmacological and molecular biological method was conducted to study the alleviating mechanism of BR at low temperature in mini Chinese cabbage. The results showed that low temperature inhibited the growth of mini Chinese cabbage seedlings, as evidenced by dwarf plants and yellow leaves. Treatment with 0.05 mg/L BR and 50 µM NO donor S-nitrosoglutathione (GSNO) significantly increased the leaf area, stem diameter, chlorophyll content, dry and fresh weight and proline content. Meanwhile, the malondialdehyde (MDA) content in 0.05 mg/L BR- and 50 µM GSNO-treated leaves were significantly lower than those in other treated leaves under low-temperature conditions. In addition, BR and GSNO applications induced an increase in NO and S-nitrosothiol (SNO) levels in vivo under low-temperature stress. Similarly, spraying BR after the elimination of NO also increased the level of S-nitrosylation in vivo, while spraying GSNO after inhibiting BR biosynthesis decreased the level of NO and SNO in vivo. In contrast, the S-nitrosoglutathione reductase (BrGSNOR) relative expression level and GSNOR enzyme activity were downregulated and inhibited by BR treatment, GSNO treatment and spraying BR after NO clearance, while the relative expression level of BrGSNOR was upregulated and GSNOR enzyme activity was also increased when spraying GSNO after inhibiting BR synthesis. Meanwhile, the biotin switch assay showed that exogenous BR increased the level of total nitrosylated protein in vivo under low-temperature stress. These results suggested that BR might act as an upstream signal of NO, induced the increase of NO content in vivo and then induced the protein S-nitrosylation modification to alleviate the damage of mini Chinese cabbage seedlings under low-temperature stress.

Lower chromosomal abnormality frequencies in miscarried conceptuses from frozen blastocyst transfers in ART.

STUDY QUESTION: Are blastocyst culture and cryopreservation in ART associated with chromosomal abnormalities in miscarried products of conception (POC)? SUMMARY ANSWER: Frozen blastocyst transfer in women aged 35 years or older and frozen embryo transfer (ET) (including both cleavage-stage embryo and blastocyst) in women aged <35 years are associated with decreased frequencies of embryonic chromosomal abnormalities in miscarried POC. WHAT IS KNOWN ALREADY: Blastocyst culture and embryo cryopreservation have been previously associated with favorable ART treatment outcomes and widely applied in clinical practice. However, the association between these embryo manipulation procedures and embryonic chromosomal abnormalities has not been evaluated to the best of our knowledge. STUDY DESIGN, SIZE, DURATION: This retrospective study included a total of 720 patients who underwent IVF/ICSI, and the retained POC were obtained. A single-nucleotide polymorphism (SNP)-based chromosomal microarray analysis (CMA) of all miscarried conceptuses was performed. PARTICIPANTS/MATERIALS, SETTING, METHODS: This study was based on the Clinical Reproductive Medicine Management System/Electronic Medical Record Cohort Database (CCRM/EMRCD) at our center. In total, 720 miscarried POCs were collected from patients undergoing ART (including fresh cleavage-stage ET, fresh blastocyst transfer, frozen cleavage-stage ET and frozen blastocyst transfer), and the incidences and profiles of cytogenetic abnormalities in the miscarried conceptuses were measured via SNP-based CMA. MAIN RESULTS AND THE ROLE OF CHANCE: The chromosomal abnormality rate in POC varied from 33.7% to 66.7% among the different ET strategies. In the patients aged ≥35 years, frozen blastocyst transfer was significantly associated with a lower incidence of chromosomal aberrations in the POCs (adjusted odds ratio (aOR): 0.171 (95% CI: 0.040-0.738); P = 0.018) than fresh blastocyst transfer. In the patients aged <35 years, frozen ET was significantly associated with a lower incidence of chromosomal aberrations than fresh ET in both cleavage-stage ET cycles and blastocyst transfers cycles (aOR: 0.545 (0.338-0.879), P = 0.013; and aOR: 0.357 (0.175-0.730), P = 0.005, respectively). Trisomy was the most frequent abnormal embryonic karyotype in the different ET strategies, and its frequency significantly differed among strategies (P < 0.05). LIMITATIONS, REASONS FOR CAUTION: This study was retrospectively designed, and we cannot draw any definite conclusions from our results regarding the adequate safety of embryo cryopreservation in ongoing pregnancy. WIDER IMPLICATIONS OF THE FINDINGS: To our knowledge, this is the first study assessing the associations of ET strategies with the probability of miscarriage associated with embryonic chromosomal abnormalities. However, the underlying mechanism of these associations is unknown; this study may promote research concerning ET strategies and promote comprehensive consultations and recommendations for patients. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the Natural Science Foundation of China (Grant No.81571409), Science and Technology Research Project of Henan (Grant No. 172102310009) and Medical Science and Technology Research Project of Henan (Grant No. 201701005). The authors declare no competing interests. TRIAL REGISTRATION NUMBER: N/A.

Patients with IVF complicated by moderate-to-critical OHSS experience increased thrombosis, GDM and neonatal NICU admission but slightly shorter gestation compared with matched IVF counterparts: A retrospective Chinese cohort study.

BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is a common disease during controlled ovarian hyperstimulation treatment. However, the obstetric and neonatal outcomes of this group of patients are unknown. The aim of this study was to explore the effects of late moderate-to-critical OHSS on obstetric and neonatal outcomes. METHODS: This prospective observational study included 17,537 patients who underwent IVF/ICSI-fresh embryo transfer (ET) between June 2012 and July 2016 and met the inclusion criteria, including 7,064 eligible patients diagnosed with clinical pregnancy. Ultimately, 6,356 patients were allocated to the control group, and 385 patients who were hospitalized and treated at the center for late moderate-to-critical OHSS were allocated to the OHSS group. Then, propensity score matching analysis was performed, matching nine maternal baseline covariates and the number of multiple gestations; 385 patients with late moderate-to-critical OHSS were compared with a matched control group of 1,540 patients. The primary outcomes were the live birth rate, preterm delivery rate, miscarriage rate, gestational age at birth (weeks), obstetric complications and neonatal complications. RESULTS: The duration of gestation in the matched control group was significantly higher than that in the OHSS group. The live birth delivery rate did not significantly differ between the OHSS and matched control groups. The incidence rates of the obstetric complications venous thrombosis (VT) and gestational diabetes mellitus (GDM), neonatal complications and the number of neonates admitted to the NICU were significantly higher in the OHSS group than in the matched control group. CONCLUSIONS: Pregnant women undergoing IVF with fresh ET whose course is complicated by late moderate-to-critical OHSS appear to experience shortened gestation and increased obstetrical and neonatal complications compared with matched controls whose course is not complicated by OHSS. However, the live birth rate, average neonatal weight, and incidence rates of premature delivery, miscarriage, early abortion, hypertensive disorder of pregnancy (HDP), placenta previa (PP), intrahepatic cholestasis of pregnancy (ICP), and low neonatal birth weight (LBW) did not differ significantly between the two groups.

Effect of progesterone on hCG day-to-basal progesterone ratio on live birth rate in long agonist fresh IVF/ICSI cycles: a 5-year, single-center study of more than 10,000 cycles.

OBJECTIVE: To investigate whether the ratio of the serum progesterone level on the day of human chorionic gonadotrophin (hCG) administration to the basal serum progesterone level (PhCG/Pbasal) is a predictor of pregnancy outcome during in vitro fertilization (IVF). METHODS: A total of 12,708 cycles were performed in 9747 patients between 19 and 36 years of age who were undergoing controlled ovarian stimulation from October 2011 to July 2016 for their first or second attempts at IVF followed by fresh embryo transfer (ET). hCG was administered 36 h before oocyte retrieval to trigger final oocyte maturation. The serum progesterone level was measured on menstrual cycle days 2-4 (basal progesterone, Pbasal) and on the day of hCG administration (PhCG). PhCG/Pbasal was calculated. Live birth rates were compared among various ordinal PhCG/Pbasal intervals (< 0.5, 0.5-1.0, 1.0-1.5, 1.5-2.0, 2.0-2.5, > 2.5). RESULTS: The average age of the patients recruited was 29.9 years. The average basal progesterone level was 0.8 ng/ml, while the average progesterone level on the day of hCG administration was 0.9 ng/ml. The live birth rates (according to the abovementioned ordinal PhCG/Pbasal intervals) were 47.3, 49.9, 47.8, 46.3, 45.5 and 44.0%, respectively. The live birth rates were significantly higher for patients with PhCG/Pbasal between 0.5-1.0 (OR = 1.14, 95% CI, 1.02-1.27, p = .02). CONCLUSIONS: PhCG/Pbasal between 0.5-1.0 predicts a higher live birth rate in IVF. Both PhCG/Pbasal and P on hCG day is less predictive value for predicting live birth rate.

Ambient air pollutant exposure and in vitro fertilization treatment outcomes in Zhengzhou, China.

OBJECTIVE: To study the association between ambient air pollutant exposure during the follicular phase and in vitro fertilization (IVF) outcomes. DESIGN: A single-center retrospective analysis. SETTING: Henan Province, China. PATIENTS: Patients (n = 6659) living in Zhengzhou, Henan Province in central China who underwent their first IVF cycle at the First Affiliated Hospital of Zhengzhou University between 2013 and 2019 were included for analysis. INTERVENTION: None. MAIN OUTCOME MEASURE: The relationships between PM2.5, PM10, and AQI (Air Quality Index) with IVF outcomes during the follicular phase (period I, 85 days before oocyte retrieval; period II, gonadotrophin start to oocyte retrieval). RESULTS: Compared with the bottom tertile, exposure to the top PM2.5 and PM10 tertiles during period I was associated with decreased clinical pregnancy (PM2.5: adjusted odds ratio [OR], 0.838%, and 95% confidence interval [CI], 0.723 and 0.971; PM10: adjusted OR, 0.818%, and 95% CI, 0.705 and 0.950), and decreased live birth rate (PM2.5: adjusted odds ratio [OR], 0.852%, and 95% confidence interval [CI], 0.736 and 0.987; PM10: adjusted OR, 0.850%, and 95% CI, 0.733 and 0.986), and exposure to the top PM2.5 tertile during period II adversely affected clinical pregnancy and the live birth rate (adjusted OR, 0.824%, and 95% CI, 0.711 and 0.955; adjusted OR, 0.817%, and 95% CI, 0.706 and 0.945). Compared with the bottom PM10 tertile, exposure to the middle PM10 tertile in period II showed decreased clinical pregnancies and live births (adjusted OR, 0.844; 95% CI, 0.729 and 0.978, adjusted OR, 0.846; 95% CI, 0.731 and 0.979). The PM10 level during period II of the follicular phase tend to adversely affect live birth rate, but the tendency did not reach significance (P = 0.051). CONCLUSION: Exposure to PM2.5 and PM10 before oocyte retrieval has an adverse effect on IVF outcomes. CAPSULE: Exposure to PM2.5 and PM10 before oocyte retrieval has an adverse effect on IVF outcomes.

Nitric oxide is involved in the brassinolide-induced adventitious root development in cucumber.

BACKGROUND: Brassinolide (BR), as a new type of plant hormones, is involved in the processes of plant growth and stress response. Previous studies have reported the roles of BR in regulating plant developmental processes and also response tolerance to abiotic stresses in plants. The main purpose of our study was to explore whether nitric oxide (NO) plays a role in the process of BR-induced adventitious root formation in cucumber (Cucumis sativus L.). RESULTS: Exogenous application of 1 µM BR significantly promoted adventitious rooting, while high concentrations of BR (2-8 µM) effectively inhibited adventitious rooting. NO donor (S-nitroso-N-acerylpenicillamine, SNAP) promoted the occurrence of adventitious roots. Simultaneously, BR and SNAP applied together significantly promoted adventitious rooting and the combined effect was superior to the application of BR or SNAP alone. Moreover, NO scavenger (c-PTIO) and inhibitors (L-NAME and Tungstate) inhibited the positive effects of BR on adventitious rooting. BR at 1 µM also increased endogenous NO content, NO synthase (NOS-like) and Nitrate reductase (NR) activities, while BRz (a specific BR biosynthesis inhibitor) decreased these effects. In addition, the relative expression level of NR was up-regulated by BR and SNAP, whereas BRz down-regulated it. The application of NO inhibitor (Tungstate) in BR also inhibited the up-regulation of NR. CONCLUSION: BR promoted the formation of adventitious roots by inducing the production of endogenous NO in cucumber.