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.

Influence of endometrial thickness on treatment outcomes following in vitro fertilization/intracytoplasmic sperm injection.

BACKGROUND: The study was designed to investigate the roles of endometrial thickness (EMT) at the day of human chorionic gonadotropin (hCG) administration on pregnancy outcomes in a large patient population. METHODS: This retrospective cohort study included 9,952 patients undergoing their first IVF/ICSI with autologous oocytes from January 2011 to January 2015. Patients were divided into three groups based on the EMT (group A:≤8 mm; group B: 9-14 mm and group C:≥15 mm). Live birth rate (LBR), clinical pregnancy rate (CPR), early miscarriage rate (EMR), and ectopic pregnancy rate (EPR) were analyzed. Additionally, the live birth rate was analyzed for patients with single or double gestational sacs. RESULTS: Significant differences (p < 0.05) were detected in the LBRs (30.38%, 45.73% and 54.55% for groups A, B, and C, respectively), CPRs (38.57%, 55.04% and 64.32%, respectively), and EPRs (5.58%, 3.48% and 2.19%, respectively), with thicker endometrial thickness favoring all three parameters. However, no differences were found in the EMRs among the three groups (15.64%, 13.44% and 13.05%, respectively, p > 0.05). After adjusting for female age, body mass index (BMI) and endometrial pattern, the multivariate logistic regression analysis demonstrated that the associations between EMT and LBR (adjusted OR: 2.645; 95% CI 2.020-3.464; p < 0.01), CPR (adjusted OR 2.693 95% CI 2.012-3.605 p < 0.01), and EPR (adjusted OR: 0.298 95% CI 0.101-0.713; p < 0.05) were significant. Additionally, live birth rates in the double gestational sac group were different (p < 0.05) among patients with different EMT (72.73%, 87.28%, and 87.36%, respectively), whereas no difference was found in the single gestational sac group. In the double gestational sac group, LBR was positively correlated with increasing endometrial thickness only in patients with twin pregnancies but not in patients with singletons. CONCLUSIONS: Our study shows that endometrial thickness at the day of hCG administration has an effect on LBR, CPR and EPR, with all three parameters increasing with the EMT. Furthermore, successful twin pregnancies are associated with a thicker endometrium.

[MicroRNA expression and its role in the cell cycle regulation in decidualized endometrial stromal cells in vitro].

OBJECTIVE: To study microRNA (miRNA) expression and role of cell cycle regulation in decidualized endometrial stormal cells (ESC) in vitro. METHODS: ESC was induced decasualization in vitro and matched with non-decidualized cells as controls. The expression repertoire of miRNA was measured by microarray chip and was validated by real-time PCR. Flow cytometry was used to identify ESC cycle during decidual reaction in vitro and after miRNA222 inhibitor was transfected into it. RESULTS: (1) Between decidualized and undecidualized stromal cells, there were 49 miRNAs significantly different expression by microarray chip, including 16 miRNA up-regulation and 33 miRNA down-regulation.hsa-miR-27b, 30c, 143, 101, 181b, 29b, 30d, 507, 23a, 222, 221 exhibited significantly differential expression between decicualized and undecidualized stromal cells by real-time PCR (P < 0.05). (2) After miRNA222 inhibitor (NC-FAM) transfection to decidual ESC, ESC were cultured by FBS medium for 24 hours, the rate of transfection was 70%. ESC were transfected with miRNA 222 inhibitor and cultured for 48 hours, the percentage of ESC at S-phase of (6.2 ± 0.7)% were significantly lower than (10.9 ± 0.8)% in control group (P < 0.05);the percentage of ESC at G(0)/G(1) phase increased at transfection group [(77.5 ± 1.3)% vs. (73.0 ± 1.6)% at control group], but there was no significant difference (P > 0.05). Decasualization ESC were transfected with miRNA 222 inhibitor and cultured for 48 h, the percentage of ESC at S-phase was (3.3 ± 0.6)% in transfection group, which were significantly lower than (7.8 ± 0.9)% in control group (P < 0.05). The percentage of ESC at G(0)/G(1) phase was (80.7 ± 1.6)% in transfection group and (74.9 ± 1.1)%. In control group, which did not reached statistical difference (P > 0.05). CONCLUSION: miRNA was involved in ESC decidual process in vitro by regulating cell cycle.

Nuclear receptor coactivator 6 promotes HTR-8/SVneo cell invasion and migration by activating NF-κB-mediated MMP9 transcription.

OBJECTIVES: NCOA6 is a transcription coactivator; its deletion in mice results in growth retardation and lethality between 8.5 and 12.5 dpc with defects in the placenta. However, the transcription factor(s) and the mechanism(s) involved in the function of NCOA6 in placentation have not been elucidated. Here, the roles of NCOA6 in human cytotrophoblast invasion and migration were studied. MATERIALS AND METHODS: Human placenta tissues were collected from normal pregnancies and pregnancies complicated by early-onset severe preeclampsia (sPE). Immunofluorescence, RT-qPCR and Western blotting were used to determine NCOA6 expression. Transwell invasion/migration assays were performed to explore whether NCOA6 knockdown affected human placenta-derived HTR-8/SVneo cell invasion/migration. Gelatin zymography was performed to examine the change in the gelatinolytic activities of secreted MMP2 and MMP9. Luciferase reporter assays were used to explore whether NCOA6 coactivated NF-κB-mediated MMP9 transcription. RESULTS: NCOA6 is mainly expressed in the human placental trophoblast column, as well as in the EVTs. HTR-8/SVneo cell invasion and migration were significantly attenuated after NCOA6 knockdown, and the secretion of MMP9 was decreased due to transcriptional suppression. NCOA6 was further found to coactivate NF-κB-mediated MMP9 transcription. Moreover, expression of NCOA6 was impaired in placentas of patients complicated by early-onset sPE. CONCLUSIONS: Thus, we demonstrated that NCOA6 is important for cytotrophoblast invasion/migration, at least partially, by activating NF-κB-mediated MMP9 transcription; the downregulation of NCOA6 may contribute to the pathogenesis of early-onset sPE.

Factors predicting clinical pregnancy rate of in vitro fertilization-embryo transfer (a STROBE-compliant article).

The aim of this study was to investigate the factors predicting clinical pregnancy rate of in vitro fertilization-embryo transfer (IVF-ET).The data of 9960 patients receiving IVF-ET fresh cycle at our Reproductive Center from January 2009 to December 2017 were first divided into pregnant group and non-pregnant group to find the clinical pregnancy rate-related factors. According to the serum HCG levels at 36 hours and 12 hours after HCG trigger, all patients were divided into 4 groups including <50 mIU/ml, ≥50 and <100 mIU/ml, ≥100 and <200 mIU/ml, and ≥200 mIU/ml groups to know whether the HCG levels at 36 hours and 12 hours affect the pregnancy rate. According to the serum HCG ratio at 36 hours to 12 hours (36 h/12 h) after HCG trigger, all patients were divided into three groups including <0.88, 0.88-1.06 and >1.06 groups to observe whether the serum HCG ratio (36 h/12 h) affects the clinical pregnancy rate. According to different assisted pregnancy modes, all patients were divided into 3 groups including IVF, ICSI, and IVF/ICSI groups to observe whether the assisted pregnancy mode affects the clinical pregnancy rate. The correlation of the clinical pregnancy rate with pregnancy rate-related factors obtained above was analyzed using logistic regression analysis model.The clinical pregnancy rate significantly increased (P < .01) in the HCG ratio (36 h/12 h) >1.06 group as compared with the HCG ratio (36 h/12 h) < 0.88 and 0.88-1.06 groups. The serum estrogen (E2) level at 36 hours was significantly lower and the number of retrieved oocytes was significantly higher in the HCG ratio (36 h/12 h) >1.06 group than in the HCG ratio (36 h/12 h) <0.88 and 0.88-1.06 groups (P = .000).The serum HCG ratio (36 h/12 h) may be used as a predictor of IVF-ET clinical pregnancy rate. High clinical pregnancy rate is probably associated with E2 down-regulation in the HCG ratio (36 h/12 h) >1.06 group.

Impact of Female Obesity on Cumulative Live Birth Rates in the First Complete Ovarian Stimulation Cycle.

Background: Female overweight/obesity has been reported to be associated with compromised pregnancy outcomes in fresh embryo transfer cycles. It is unclear whether the cumulative live birth rate (CLBR) is adversely affected after all viable embryos are transferred from the first ovarian stimulation cycle. Objectives: To investigate whether the CLBR was compromised in obese women. Method: A total of 9,772 young women underwent their first IVF/ICSI cycles from January 2012 to October 2017. Pregnancy outcomes were compared according to female BMI. Results: Among 1,671 women with polycystic ovary syndrome (PCOS), those with a BMI ≥ 28 kg/m2 had a lower cumulative clinical pregnancy rate (CCPR) and CLBR during the first complete ovarian stimulation cycle. Additionally, the pregnancy loss rate was increased in this group, although the difference was not significant. Among the 8,101 women without PCOS, the CCPR and CLBR of obese patients was also significantly decreased, and this group also showed increased pregnancy loss rates. Moreover, overweight women also had a decreased CLBR. Conclusions: Female obesity adversely affected the CLBR after utilizing the viable embryos from first oocytes retrieval.

miR-181a-5p suppresses invasion and migration of HTR-8/SVneo cells by directly targeting IGF2BP2.

Pre-eclampsia is a pregnancy-related disease that may cause maternal, neonatal and fetal morbidity and mortality and exists in 3-5% of pregnancies worldwide. The discovery of dysregulated microRNAs and their roles in placental development has provided a new avenue for elucidating the mechanism involved in this pregnancy-specific disorder. Here, the roles of human miR-181a-5p, a microRNA that is increased in both the plasma and placenta of severe pre-eclamptic patients, in invasion and migration of trophoblasts were investigated. Ectopic-expression of miR-181a-5p impaired the invasion and migration of HTR-8/SVneo cells, whereas miR-181a-5p inhibition had the opposite effects. IGF2BP2, which harbors a highly conserved miR-181a-5p-binding site within its 3'-UTR, was identified to be directly inhibited by miR-181a-5p. Moreover, siRNAs targeting IGF2BP2 imitated the effects of overexpressed miR-181a-5p on HTR-8/SVneo cell invasion and migration, whereas restoring IGF2BP2 expression by overexpressing a plasmid encoding IGF2BP2 partially reversed the studied inhibitory functions of miR-181a-5p. Thus, we demonstrated here that miR-181a-5p suppresses the invasion and migration of cytotrophoblasts, and its inhibitory effects were at least partially mediated by the suppression of IGF2BP2 expression, thus shedding new light on the roles of miR-181a-5p in the pathogenesis of severe pre-eclampsia.

[Effects of Ca2+ on nitric oxide-induced adventitious rooting in cucumber under drought stress]. / å¹²æ—±æ¡ä»¶ä¸‹é’™ç¦»å­å¯¹ä¸€æ°§åŒ–æ°®è¯±å¯¼é»„ç“œä¸å®šæ ¹å‘ç”Ÿçš„å½±å“.

Cucumber (Cucumis sativus L. 'Xinchun 4') was used to explore the relationship between nitric oxide (NO) and calcium (Ca2+) during adventitious rooting under drought stress. Rooting parameters, endogenous Ca2+ fluorescent intensity and the antioxidant enzymes activity (SOD, CAT and APX) in cucumber explants under drought stress were investigated. The results showed that treatment with 200 µmol·L-1 CaCl2 and 0.05% PEG significantly improved the number and length of adventitious root in cucumber explants under drought stress, while the application of Ca2+ chelating agent (EGTA) and channel inhibitor (BAPTA/AM) significantly decreased NO-induced number and length of adventitious root under drought stress. Under drought stress, the fluorescence intensity of Ca2+ in hypocotyls treated with NO and CaCl2 was improved, however, the Ca2+ fluorescence intensity in the hypocotyls treated with NO scavenger (cPTIO) was significantly lower than that in the hypocotyls treated with NO. Under drought stress, the activities of antioxidant enzymes in the cucumber explants were significantly promoted by the treatments with NO and CaCl2, however, Ca2+ chelating agent and channel inhibitor significantly decreased the activity of antioxidant enzymes induced by NO. In conclusion, Ca2+ might be involved in the process of NO-adjusted antioxidant enzymes activity during adventitious rooting under drought stress, which alleviated the negative effects of drought on the adventitious rooting and promoted the formation of adventitious roots.

[Effects of exogenous salicylic acid and brassinolide on photosynthesis of cucumber seedlings under low temperature stress.] / æ°´æ¨é ¸å’Œæ²¹èœç´ å† é ¯å¯¹ä½Žæ¸©èƒè¿«ä¸‹é»„ç“œå¹¼è‹—å ‰åˆä½œç”¨çš„å½±å“.

An experiment was conducted to investigate the mechanism of the regulation of salicylic acid (SA) and 2,4-epibrassionolide (EBR) on photosynthesis in cucumber (Cucumis sativa) seedlings under low temperature stress. Taking cucumber cultivar 'Youbo1-5' as material, the seedlings were pre-treated with 1 mmol·L-1 SA or 0.1 µmol·L-1EBR (sprayed once a day), and then were exposed to chilling temperature (10 ℃/5 ℃, PFD 80 µmol·m-2·s-1) after being pre-treated 2 days. The results showed that the growth and net photosynthetic rate (Pn) of cucumber seedlings were decreased under low temperature stress. However, the Pn, maximal photochemical efficiency of PS2 (Fv/Fm), actual photochemical efficiency of PS2 (ΦPS2) and photochemical quenching coefficient (qP) were significantly improved in SA- and EBR-pretreated seedlings, and the increase range of non-photochemical quenching coefficient (NPQ) was decreased. Moreover, the activities of ribulose 1,5-biphosphate carboxylase/oxygenase (Rubisco), sedoheptulose-1,7-bisphosphatase (SBPase), fructose-1,6-bisphosphate aldolase (FBA) and transketolase (TK) were signi-ficantly increased. These findings suggested that SA and EBR improved photosynthesis of cucumber seedlings by promoting the activities of key enzymes and increased low temperature tolerance.

Molecular karyotype single nucleotide polymorphism analysis of early fetal demise.

We explored the application of single nucleotide polymorphism microarray (SNP array) in molecular karyotype analysis for early spontaneous abortion detection in assisted reproductive technology (ART). SNP array was performed in 81 cases. Of the 81 cases, 16 experienced natural conception (NC) and 65 were pregnant by ART. Of the 65 cases, 4 underwent artificial insemination (AI), 32 fresh in vitro fertilization-embryo transfer (IVF-ET), 9 fresh intracytoplasmic sperm injection (ICSI), and 20 thawed embryo transfer. In the 81 cases examined 69.1% displayed an abnormal molecular karyotype. In the subjects greater than 35 years of age, the abnormal molecular karyotype rate was 87.5% higher compared to 61.4% in younger individuals (P < 0.05). There was no significant difference in the abnormal molecular karyotype rate or type between ART (64.6%) and NC (87.5%). Compared with traditional cytogenetic diagnosis, the SNP array can identify a greater number of abnormal karyotypes.

Relationship between processed total motile sperm count of husband or donor semen and pregnancy outcome following intrauterine insemination.

We retrospectively analyzed 6,360 artificial insemination cycles of husband's semen through intrauterine insemination (AIH-IUI) or artificial insemination with donor semen through intrauterine insemination (AID-IUI) in patients with infertility between August, 1998 and August, 2010. The relationship between processed total motile sperm count (PTMS) and pregnancy outcome was determined. The study was divided into 6 groups according to PTMS. Group 1: ≤ 2.0 million, Group 2: 2.1-4.0 million, Group 3: 4.1-6.0 million, Group 4: 6.1-8.0 million, Group 5: 8.1-10.0 million, and Group 6: >10.0 million. There was no statistically significant difference in age, duration of infertility, unilateral tubal patency, induced ovulation, and single IUI or double IUI between the 6 groups in both AIH-IUI and AID-IUI. The total clinical pregnancy rate of AIH-IUI was 10.81 % and AID-IUI was 27.52 %. Among the 6 groups, the clinical pregnancy rate was the lowest in Group 1 (P < 0.05) in both AIH-IUI and AID-IUI. With the increased PTMS, the clinical pregnancy rate of IUI was improved. However, a statistical difference between groups was only observed for Group 1. When PTMS is ≤ 2 × 10(6) the clinical pregnancy rate of IUI is significantly decreased. In this case in vitro fertilization (IVF) should be adopted.