The effect of an extended culture period on birth weight among singletons born after single or double vitrified embryo transfer.

Aim: To evaluate the effect of an extended culture period on birth weight among singletons born after vitrified-warmed embryo transfer. Methods: A retrospective cohort study was performed among 12400 women who gave birth to 1015, 1027, 687, and 9671 singletons after single blastocyst transfer, single cleavage-stage embryo transfer, double blastocyst transfer, and double cleavage-stage embryo transfer, respectively. Results: The unadjusted birth weight of singletons born after vitrified blastocyst transfer were heavier than those born after cleavage-stage transfer (ß=30.28, SE=13.17, P=0.022), as were the adjusted birth weights (ß=0.09, SE=0.03, P=0.007). In addition, there was a 37% increased odd of having an infant with high birth weight after vitrified blastocyst transfer compared with vitrified cleavage stage transfer (OR=1.37, 95% CI:1.07-1.77). Conclusion: The unadjusted and adjusted birth weight and odds of having an infant with high birth weight significantly increased after blastocyst transfer compared with cleavage-stage embryo transfer in vitrified-warmed cycles.

The impact of blastomere loss on pregnancy and neonatal outcomes of vitrified-warmed Day3 embryos in single embryo transfer cycles.

BACKGROUND: Blastomere loss is a common phenomenon that occurs following cryopreservation. To date, studies have drawn conflicting conclusions regarding the impact of blastomere loss on pregnancy outcomes. Besides, limited information is available concerning the neonatal safety of embryos with blastomere loss. In the present study, we aimed to investigate the impact of blastomere loss on pregnancy and neonatal outcomes of vitrified/warmed Day3 cleavage-stage embryos in single embryo transfer cycles. METHODS: This retrospective cohort study included all vitrified/warmed D3 cleavage-stage single frozen-thawed embryo transfer (FET) cycles between April 2015 and February 2021. We compared pregnancy and subsequent neonatal outcomes between the intact embryos group and the blastomere loss group in single FET cycles. RESULTS: A total of 6287 single FET cycles were included in the study, in which 5873 cycles were classified into the intact embryo group and 414 cycles were classified into the blastomere loss group. The outcomes of the blastomere loss group were significantly inferior to those of the intact embryo group, in terms of implantation/biochemical pregnancy/clinical pregnancy/ongoing pregnancy rate and live birth rate per embryo transfer cycle/per clinical pregnancy. Further binary logistic regression confirmed that blastomere loss was negatively associated with live birth. Moreover, the blastomere loss group presented with an elevated early miscarriage rate. The neonatal conditions were broadly similar between the two groups. Additionally, multiple binary logistic regression analysis demonstrated that primary infertility and intracytoplasmic sperm injection (ICSI) were common influencing factors of blastomere loss (aOR 1.447, 95% CI 1.038-2.019, P = 0.029; aOR: 1.388, 95% CI: 1.044-51.846, P = 0.024). CONCLUSIONS: The transfer of vitrified/warmed D3 embryos with blastomere loss is related to impaired embryo developmental potentials and reduced probabilities of conception. Moreover, even if the embryos with blastomere loss have implanted and reached clinical pregnancies, they present with a lower possibility of developing to live birth owing to a higher early miscarriage rate. However, once the embryos with blastomere loss result in a live birth, no adverse neonatal outcomes are observed. Primary infertility and ICSI were found to be risk factors for blastomere loss.

The Effect of Spontaneous LH Surges on Pregnancy Outcomes in Patients Undergoing Letrozole-HMG IUI: A Retrospective Analysis of 6,285 Cycles.

Background: To date, no consensus has been reached on whether to wait for spontaneous luteinizing hormone (LH) surge to occur or to trigger ovulation regardless of the presence of an LH surge for achieving higher success rate in intrauterine insemination (IUI) cycles. Therefore, we hope to investigate the effect of the presence of a spontaneous LH surge on pregnancy outcomes in letrozole-human menopausal gonadotropin (LE-HMG) IUI cycles. Methods: In this retrospective cohort study, a total of 6,285 LE-HMG IUI cycles were included between January 2010 and May 2021. Cycles were categorized into three groups: the trigger + LH surge group, the trigger only group, and the LH surge only group. The primary outcome measure was the clinical pregnancy rate. A logistic regression analysis was performed to explore other risk factors affecting the clinical pregnancy rate. Results: No significant differences were observed in biochemical pregnancy rate (P =0.640), clinical pregnancy rate (P =0.702), ongoing pregnancy rate (P =0.842), and live birth rate (P =0.951) among the three groups. The binary logistic regression analysis also confirmed that the existence of an LH surge was not associated with clinical pregnancy. There was a difference in ectopic pregnancy rates (P =0.045), but logistic regression showed that the presence of a spontaneous LH surge has no association with ectopic pregnancy. Nonetheless, patients with lead follicles within 18.1-20.0 mm/20.1-22.0 mm and a long duration of LE treatment were less likely to get ectopic pregnant compared with patients with 14.1-16.0 mm lead follicles and shorter LE treatment (OR: 0.142, 95% CI: 0.023-0.891, P =0.037; OR: 0.142, 95% CI: 0.022-0.903, P =0.039; OR: 0.445, 95% CI: 0.235-0.840, P = 0.013). Conclusions: The presence of a spontaneous LH surge in triggered LE-HMG IUI cycles does not appear to improve pregnancy rates. Thus, we suggest that waiting for an LH surge to occur is not necessary in triggered LE-HMG IUI cycles.

Nintedanib Treatment After Ovulation is an Effective Therapeutic Strategy for the Alleviation of Ovarian Hyperstimulation Syndrome (OHSS) in a Mouse Model.

PURPOSE: Ovarian hyperstimulation syndrome (OHSS) is a serious complication of controlled ovarian hyperstimulation. In this study, we hope to explore whether nintedanib, a tyrosine kinase inhibitor, can inhibit OHSS by blocking signaling of vascular endothelial growth factor in a mouse model. Considering that nintedanib been approved for the treatment of some diseases. We believe that nintedanib has important potential in the treatment of OHSS. METHODS: Female ICR mice aged 6-8 weeks with similar initial weights were used to establish the OHSS model. At 12 and 24 hours after human chorionic gonadotropin (hCG) trigger, we administered nintedanib by subcutaneous injection and analyzed the OHSS-related physiological characteristics and biochemical indices of the model mice within 48 hours after hCG-trigger. RESULTS: Nintedanib significantly alleviated the symptoms of OHSS after hCG-trigger compared with those of OHSS group (weight change, P < 0.0001; ovarian weight, P < 0.0001, peritoneal exudation level, P < 0.01). Further investigation proved that the corpus luteum (number, P < 0.001; diameter, P < 0.0001) and luteal vessel (P < 0.0001) development were inhibited in the nintedanib administration group. Then, the vascular permeability test showed that the capillary bleeding points (P < 0.0001) were also significantly reduced in nintedanib administration group. Gene expression tests demonstrated that the intercellular connection-related genes expression in the nintedanib administration group was similar to that in the no-OHSS induced group. Further detection of coagulation and thrombosis indices indicated that the nintedanib administration in the OHSS model did not increase the risk of thrombosis or bleeding. CONCLUSION: Our study demonstrated that nintedanib can alleviate and manage the symptoms of OHSS in a mouse model. These findings identify a feasible scheme for the prevention and treatment of OHSS in clinical practice in the future. Moreover, since the scheme can be implemented after ovulation, it will not cause potential adverse effects on gametogenesis, fertilization or embryonic development.

YTHDF2 is essential for spermatogenesis and fertility by mediating a wave of transcriptional transition in spermatogenic cells.

The dynamic and reversible regulation roles of m6A modification and the characterization of m6A readers have provided new insights into spermatogenesis at the post-transcriptional level. YTHDF2, as an m6A reader, has been reported to mediate the m6A-containing transcript decay during the mouse oocyte maturation, embryonic stem cell differentiation, neural development, and zebrafish maternal-to-zygotic transition. However, the roles of YTHDF2 in mammalian spermatogenesis are uncertain. Here, we generated germ cell-specific Ythdf2 mutants (Ythdf2-vKO) at a C57BL/6J background and demonstrated that YTHDF2 is essential for mouse spermatogenesis and fertility. Ythdf2-vKO provides oligoasthenoteratozoospermia phenotype with increased apoptosis in germ cells. High-throughput RNA-seq analysis showed that a group of mRNAs is upregulated in Ythdf2-vKO mouse testis; further analysis and MeRIP-qPCR data showed that most of the upregulated genes in Ythdf2-vKO mouse testis are modified with m6A and are YTHDF2 candidate binding genes. Interestingly, RNA-seq analysis combined with our previous single-cell transcriptomics data of mouse spermatogenesis pointed out the failure of a wave of transcript transition during the spermatogenesis of Ythdf2-vKO mice, which was confirmed by gene expression analysis using qPCR of diplotene spermatocytes and round spermatids obtained through fluorescence-activated cell sorting. Our study demonstrates the fundamental role of YTHDF2 during mouse spermatogenesis and provides a potential candidate for the diagnosis of male infertility with the oligoasthenoteratozoospermia syndrome.

Effect of Day 3 and Day 5/6 Embryo Quality on the Reproductive Outcomes in the Single Vitrified Embryo Transfer Cycles.

Objective: To explore the live birth rate and neonatal outcome after single vitrified blastocyst transfer versus single vitrified cleavage-stage embryo transfer at different grades of embryo quality. Methods: A retrospective cohort study including 6077 single vitrified-thawed embryo transfer cycles was performed in the time-period from January 2013 to December 2018. Results: After controlling for potential confounding variables, there are 161% increased odds of a live birth after transfer of single good quality embryo at day 5, 152% increased odds of a live birth after transfer of single poor quality embryo at day 5, 60% increased odds of a live birth after transfer of single good quality embryo at day 6 compared with transfer of single good quality embryo at day 3. Results from the generalized estimated equation regression showed significant relationship of unadjusted birth weight with development stage of embryo and embryo quality (good quality embryo on day 5 vs. Good quality embryo on day 3:ß=108.55, SE=34.89, P=0.002; good quality embryo on day 6 vs. Good quality embryo on day 3:ß=68.80, SE=33.75, P=0.041). However, no significant differences were seen in birth weight between transfer single poor quality embryo on day 5, 6 and transfer single good quality embryo on day 3. Conclusion: A significant increase in live birth rate and birth weight after transfer of single good quality embryo on day 5 and day 6 compared with transfer of single good quality embryo on day 3 in the vitrified embryo transfer cycles.

Protein Arginine Methyltransferase 6 (Prmt6) Is Essential for Early Zebrafish Development through the Direct Suppression of gadd45αa Stress Sensor Gene.

Histone lysine methylation is important in early zebrafish development; however, the role of histone arginine methylation in this process remains unclear. H3R2me2a, generated by protein arginine methyltransferase 6 (Prmt6), is a repressive mark. To explore the role of Prmt6 and H3R2me2a during zebrafish embryogenesis, we identified the maternal characteristic of prmt6 and designed two prmt6-specific morpholino-oligos (MOs) to study its importance in early development, application of which led to early epiboly defects and significantly reduced the level of H3R2me2a marks. prmt6 mRNA could rescue the epiboly defects and the H3R2me2a reduction in the prmt6 morphants. Functionally, microarray data demonstrated that growth arrest and DNA damage-inducible, α, a (gadd45αa) was a significantly up-regulated gene in MO-treated embryos, the activity of which was linked to the activation of the p38/JNK pathway and apoptosis. Importantly, gadd45αa MO and p38/JNK inhibitors could partially rescue the defect of prmt6 morphants, the downstream targets of Prmt6, and the apoptosis ratios of the prmt6 morphants. Moreover, the results of ChIP quantitative real time PCR and luciferase reporter assay indicated that gadd45αa is a repressive target of Prmt6. Taken together, these results suggest that maternal Prmt6 is essential to early zebrafish development by directly repressing gadd45αa.

[Roles of matrix metalloproteinases, tissue inhibitor of matrix metalloproteinase and hs-CRP in Kawasaki disease].

OBJECTIVE: To examine the expression of matrix metalloproteinases (MMP)-2, -9, tissue inhibitor of matrix metalloproteinase (TIMP)-1 and hs-CRP, and their relationship with coronary artery in children with Kawasaki disease. METHODS: One hundred and fifty-one children with Kawasaki disease (111 cases with coronary artery damage and 40 cases without) and 60 healthy children were enrolled. The expression of MMP-2, MMP-9 and TIMP-1 was detected using ELISA, and the hs-CRP concentration was measured using the endpoint nephelometry. RESULTS: There were significant differences in the level of MMP-2, MMP-9 and hs-CRP between the patients with or without coronary artery damage and the healthy children (p<0.05). The levels of MMP-2, MMP-9 and hs-CRP were the highest in the cardiovascular damage group (p<0.05). There were positive correlations between MMP-2, MMP-9 and TIMP-1 in children with Kawasaki disease. CONCLUSIONS: MMP-2, MMP-9, TIMP-1 and hs-CRP may play important roles in the development of Kawasaki disease. The combined measurement of MMP-2, MMP-9 and hs-CRP may be useful in the evaluation of the severity in children with Kawasaki disease.

[Choline chloride protects cell membrane and the photosynthetic apparatus in cucumber seedling leaves at low temperature and weak light].

Choline chloride 1.07 mmol/L treatment diminished the saturated lipid contents of the fatty acid components mainly the phosphatidylglycerol (PG) resulting in the decrease of the saturation of lipid (Table 1), declined the permeability of cell membrane and the production of MDA from lipid peroxidation (Fig.2) in the cucumber seedling leaves under low temperature and weak light (6 degrees C, PFD 100 micromol m(-2) s(-1)). Furthermore, the choline chloride treatment alleviated the degradation of chlorophyll pigments especially chlorophyll b, the decrease in maximum photochemical efficiency (Fv/Fm), the capture efficiency of excited energy (Fv'/Fm'), the photochemical quenching coefficient (q(p)) and the actual photochemical efficiency (Phi PSII) of PSII (Table 2, Fig.3A, B & C), and decreases in activity of antioxidant enzymes such as POD, APX and CAT (Fig.4) in chilled leaves under weak light. In addition, choline chloride treatment increased the non-photochemical quenching (NPQ) (Fig.3D) and the proline content (Fig.5) in chilled leaves under weak light. The above results indicate that choline chloride protected the cell membrane and the photosynthetic apparatus in cucumber seedling leaves from chilling stress in weak light.

[Effects of root-applied glycinebetaine on the composition and function of wheat thylakoid membrane under drought stress].

Two wheat cultivars (Triticum aestivum L.), HF9703 tolerant to drought and SN215953 sensitive to drought, were used to study the effects of glycinebetaine on the composition and function of thylakoid membrane. The wheat seedlings with two leaves were pretreated with Hoagland solution containing 1.5 mmol/L glycinebetaine (GB) for 72 h, then cultured with Hoagland solution containing 15% PEG-6000 for 48 h. The seedling leaves were used for mensuration. The results indicated that the chlorophyll, monogalactosyl diaylglycerol (MGDG), digalactosyl diacylglycerol (DGDG) and phosphatidylglycerol (PG) contents of the two wheat cultivars decreased significantly (P<0.05) under drought stress. GB alleviated their decrease. The sulfoquinovosyl diacylglycerol (SQDG) content, trans-hexadecenoic [16:1(3t)] and saturated fatty acid content of MGDG in HF9703 increased significantly (P<0.05), while in SN215953, the sulfoquinovosyl diacylglycerol (SQDG) and trans-hexadecenoic [16:1(3t)] contents decreased significantly under drought stress, but the saturated fatty acid content of MGDG increased slightly. These differences between the two wheat cultivars might account for much of the difference in drought tolerance between them. Ca2+-ATPase activity of thylakoid membrane, Hill-reaction activity and photosynthesis were decreased markedly (P<0.05) by drought stress. GB ameliorated these effects on thylakoid membrane, and the effect of GB on SN215953 was stronger than on HF9703. Discussion was made on the possible mechanism of the alleviating effect of root-applied GB on the composition and function of thylakoid membrane.