NAT10-mediated N4-acetylcytidine modification is required for meiosis entry and progression in male germ cells.

Post-transcriptional RNA modifications critically regulate various biological processes. N4-acetylcytidine (ac4C) is an epi-transcriptome, which is highly conserved in all species. However, the in vivo physiological functions and regulatory mechanisms of ac4C remain poorly understood, particularly in mammals. In this study, we demonstrate that the only known ac4C writer, N-acetyltransferase 10 (NAT10), plays an essential role in male reproduction. We identified the occurrence of ac4C in the mRNAs of mouse tissues and showed that ac4C undergoes dynamic changes during spermatogenesis. Germ cell-specific ablation of Nat10 severely inhibits meiotic entry and leads to defects in homologous chromosome synapsis, meiotic recombination and repair of DNA double-strand breaks during meiosis. Transcriptomic profiling revealed dysregulation of functional genes in meiotic prophase I after Nat10 deletion. These findings highlight the crucial physiological functions of ac4C modifications in male spermatogenesis and expand our understanding of its role in the regulation of specific physiological processes in vivo.

Revisiting ZAR proteins: the understudied regulator of female fertility and beyond.

Putative RNA-binding proteins (RBPs), zygote arrested-1 (ZAR1), and ZAR2 (also known as ZAR1L), have been identified as maternal factors that mainly function in oogenesis and embryogenesis. Despite divergence in their spatio-temporal expression among species, the CxxC structure of the C-terminus of ZAR proteins is highly conserved and is reported to be the functional domain for the activity of the RBPs of ZAR proteins. In oocytes from Xenopus laevis and zebrafish, ZAR proteins have been reported to bind to maternal transcripts and inhibit translation in immature growing oocytes, whereas in fully grown mouse oocytes, they promote the translation during meiotic maturation. Thus, ZAR1 and ZAR2 may be required for the maternal-to-zygotic transition by stabilizing the maternal transcriptome in oocytes with partial functional redundancy. In addition, recent studies have suggested non-ovarian expression and function of ZAR proteins, particularly their involvement in tumorigenesis. ZAR proteins are potentially associated with tumor suppressors and can serve as epigenetically inactivated cancer biomarkers. In this review, studies on Zar1/2 are systematically summarized, and some issues that require discussion and further investigation are introduced as perspectives.

[Pathological characteristics and prognosis of 664 patients with epithelial ovarian cancer: a retrospective analysis].

OBJECTIVE: To analyze clinicopathologic features and risk factors associated with the recurrence and prognosis of epithelial ovarian cancer (EOC). METHODS: 710 EOC patients treated at the West China University Second Hospital from Jan. 2006 to Jun. 2011 were recruited in this study retrospectively. Univariate and multivariate logistic regression models were constructed to evaluate the risk of factors. Kaplan-Meier and log-rank methods were adopted for survival analyses. RESULTS: The final sample included 664 patients with complete and well-documented data. The participants had a mean age of (49.35 +/- 11.58) yr. and 79.07% (525/664) were older than 40 year-old. CA125 was tested in 550 patients before surgery and 82.55% showed abnormal values. Over half (55.57%) of the patients were classified as serious EOC, which was followed by clear cell EOC (12.35%), endometrioid EOC (10. 09%), mucinous EOC (7.68%), and others (14.31%). Stage I (FIGO) comprised 30. 72% of the cases. The patients were followed up on average of (37.48 +/- 12.51) months and 303 died with a mean survival length of (28.54 +/- 9.56) months. Recurrence was found in 126 patients at a median of (26.10 +/- 5.75) months. For the 361 survived, 81. 72% lived without detectable cancer. All patients received surgical operations, including 483 undergoing retroperitoneal lymphadenectomy. The univariate analysis identified older age, advanced FIGO stage, suboptimal debulking, abnormal CA125 values before surgery, undifferentiated, mixed type and serous pathologic subtypes, poor-differentiation and pathogenesis of tumor as risk factors associated with survival prospect. The multivariate logistic regression models confirmed that poor prognosis was associated with older age, advanced FIGO stage, suboptimal debulking and undifferentiated, mixed type and serous pathologic subtype. CONCLUSION: Older age, advanced FIGO stage, high grade differentiation, suboptimal debulking, lymphnode metastasis, and type II EOC are associated with poor prognosis of EOC patients.

Exosomal circNFIX promotes angiogenesis in ovarian cancer via miR-518a-3p/TRIM44 axis.

Ovarian cancer (OC) is a gynecological cancer with high mortality. OC-derived exosomal circRNAs can regulate angiogenesis. This study aims to explore the role and mechanism of exosomal circRNA nuclear factor I X (CircNFIX) derived from OC cells in angiogenesis. Quantitative real-time polymerase chain reaction was employed to evaluate the levels of circNFIX, miR-518a-3p, and tripartite motif protein 44 (TRIM44) in OC and adjacent tissues. Exosomes from the ovarian surface epithelial cell (HOSEpiC) and OC cells (SKOV3 or OVCAR3) were isolated by differential centrifugation. Exosomes were cocultured with the human umbilical vein endothelial cells (HUVECs). The angiogenesis capacity was analyzed by Tube formation assay. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and Transwell assays were used to determine the cell viability and migration ability. The dual-luciferase report, RNA immunoprecipitation (RIP), and RNA pull-down assays were applied to validate the gene's interaction. CircNFIX and TRIM44 expression were higher and miR-518a-3p was lower in OC tissues than in the adjacent tissues. Upregulated circNFIX and TRIM44 were significantly correlated with the tumor size and International Federation of Gynecology and Obstetrics (FIGO) stage of OC patients. HUVECs treated OC-derived exosomes had higher proliferation, migration, and angiogenesis capacities than the control group. While OC-derived exosomal circNFIX silencing restrained HUVECs' proliferation, migration, and angiogenesis, compared with the OC-derived exosomes group. OC-derived exosomal circNFIX positively regulated TRIM44 expression by targeting miR-518a-3p in HUVECs. OC-derived exosomal circNFIX promoted angiogenesis by regulating the Janus-activated kinase/signal transducer and activator of transcription 1 (JAK/STAT1) pathway via miR-518a-3p/TRIM44 axis in HUVECs.

CircNFIX stimulates the proliferation, invasion, and stemness properties of ovarian cancer cells by enhancing SH3RF3 mRNA stability via binding LIN28B.

We aimed to study the regulatory roles and mechanism of circular nuclear factor IX (circNFIX) in cancer growth and stemness properties of ovarian cancer (OC). CircNFIX and SH3RF3 levels in OC tissues and cells were tested by quantitative real-time PCR. RNase R treatment quantified circNFIX RNA stability. Molecular interaction among circNFIX, LIN28B, and SH3RF3 was predicted by bioinformatics software and validated through RNA immunoprecipitation (RIP) assay. The gain- or loss-experiments of circNFIX on capabilities of metastasis and stemness in vitro were assessed using Cell Counting Kit-8, Transwell, western blot, and sphere-formation assays. CircNFIX and SH3RF3 were markedly elevated in OC tissues and OC cells. Knocking down circNFIX repressed the proliferation, migration, invasion, and stemness properties of A2780 and SKOV3 cells. The RIP assay verified the direct binding relationship between LIN28B, circNFIX, and SH3RF3. Additionally, overexpression of circNFIX elevated the SH3RF3 expression, while this effect was reversed by LIN28B silence. Rescue experiments demonstrated that the overexpression of SH3RF3 reversed the knockdown of circNFIX on OC cells' proliferation, metastasis, and stemness properties. CircNFIX improved the mRNA stability and translation of SH3RF3 via recruiting LIN28B, thus promoting the proliferation, invasion, and stemness properties of OC cells in vitro.

Bisphenol A Exposure Disrupts Organelle Distribution and Functions During Mouse Oocyte Maturation.

Bisphenol A (BPA) is one of the ubiquitous environmental endocrine disruptors (EEDs). Previous studies have shown that the reproduction toxicity of BPA could cause severe effects on the mammal oocytes and disturb the quality of mature oocytes. However, the toxic effects of BPA on the organelles of mouse oocytes have not been reported. In this study, to investigate whether BPA can be toxic to the organelles, we used different concentrations of BPA (50, 100, and 200 µM) to culture mouse oocytes in vitro. The results showed that 100 µM BPA exposure could significantly decrease the developmental capacity of oocytes. Then, we used the immunofluorescence staining, confocal microscopy, and western blotting to investigate the toxic effects of BPA on the organelles. The results revealed that mitochondrial dysfunction is manifested by abnormal distribution and decreased mitochondrial membrane potential. Moreover, the endoplasmic reticulum (ER) is abnormally distributed which is accompanied by ER stress showing increased expression of GRP78. For the Golgi apparatus, BPA-exposed dose not disorder the Golgi apparatus distribution but caused abnormal structure of Golgi apparatus, which is manifested by the decrease of GM130 protein expression. Moreover, we also found that BPA-exposed led to the damage of lysosome, which were shown by the increase of LAMP2 protein expression. Collectively, our findings demonstrated that the exposure of BPA could damage the normal function of the organelles, which may explain the reduced maturation quality of oocytes.

Enhancing Multiple Jets in Electrospinning: The Role of Auxiliary Electrode.

An auxiliary electrode introduced in traditional spinneret electrospinning is an effective and powerful technique to improve the production rate of nanofibers. In this work, the effects of the arrangement of auxiliary electrode, applied voltage, injection speed, and the distance between the electrode tip and the spinneret tip (ESD) on the jet number and the morphology of polyvinyl alcohol (PVA) nanofibers were investigated systematically. The results showed that the number of jets firstly increased and then decreased with the increase of applied voltage and ESD, respectively, while increasing with the injection speed in both the auxiliary electrode in the vertical position and parallel position. The average nanofiber diameter decreased with increasing of applied voltage and injection speed, but decreasing in ESD in these two positions. The numerical simulation results revealed that the auxiliary electrode primarily influenced the electric field intensity in the spinning area. This work provides a deep understanding of multiple jets in electrospinning.

Multi-Jet Electrospinning with Auxiliary Electrode: The Influence of Solution Properties.

Multiple jets ejection in electrospinning has been a major approach to achieving a high production rate of ultrafine fibers, also known as nanofibers. This work studies the effect of solution parameters-including dielectric constant, polarity, conductivity and surface tension-on the jet number and jet evolution in the auxiliary electrode electrospinning approach. The results show that it is easier to generate 2⁻6 jets with short stable jet length (1.7⁻6.9 mm) under low voltage (5.03⁻7.13 kV) when solutions have higher dielectric constant (32.2⁻78.6) and larger surface tension (31.8⁻41.29 mN/m). The influence of solution properties on stable jet length and the influence of applied voltage to produce multiple jets are discussed in detail. This work provides a new perspective for understanding jet evolution and mass production of nanofibers in electrospinning.

Hierarchical Porous Polyamide 6 by Solution Foaming: Synthesis, Characterization and Properties.

Porous polym er materials have received great interest in both academic and industrial fields due to their wide range of applications. In this work, a porous polyamide 6 (PA6) material was prepared by a facile solution foaming strategy. In this approach, a sodium carbonate (SC) aqueous solution acted as the foaming agent that reacted with formic acid (FA), generating CO2 and causing phase separation of polyamide (PA). The influence of the PA/FA solution concentration and Na2CO3 concentration on the microstructures and physical properties of prepared PA foams were investigated, respectively. PA foams showed a hierarchical porous structure along the foaming direction. The mean pore dimension ranged from hundreds of nanometers to several microns. Low amounts of sodium salt generated from a neutralization reaction played an important role of heterogeneous nucleation, which increased the crystalline degree of PA foams. The porous PA materials exhibited low thermal conductivity, high crystallinity and good mechanical properties. The novel strategy in this work could produce PA foams on a large scale for potential engineering applications.

Hyperbaric Oxygenation Protects Against Ischemia-Reperfusion Injury in Transplanted Rat Kidneys by Triggering Autophagy and Inhibiting Inflammatory Response.

BACKGROUND Ischemia-reperfusion injury (IRI) is a clinically common pathologic process defined as the inability to improve neuronal function. This study aimed to investigate the pathological mechanism of IRI and to explore effects of hyperbaric oxygenation (HBO) on autophagy and inflammatory response in IRI. MATERIAL AND METHODS Ninety Sprague-Dawley (SD) rats were randomly divided into a Sham group, a kidney transplant group (Trans), and a kidney transplant plus HBO treatment group (Trans+HBO). The kidney was harvested from the donor and transplanted to recipient rats according to a previously reported study. Rats were anesthetized using pentobarbital-natrium, and the kidney was resected and fixed in 4% paraformaldehyde. Serum creatinine (Scr) was detected using an automatic biochemical analyzer. The interleukin-6 (IL-6) level was assessed using enzyme-linked immunosorbent assay (ELISA). LC-3 was examined using indirect immunofluorescence assay and immunochemistry assay. LC-3 mRNA levels were analyzed using real-time PCR (RT-PCR). RESULTS The kidney transplant IRI model was successfully established. Scr and IL-6 levels were significantly increased in the Trans group (P<0.05). HBO significantly enhanced Scr and IL-6 levels. Scr was positively correlated with IL-6 levels (r-0.607, P<0.05). HBO increased LC-3 protein and mRNA expression in kidney-transplanted rats compared to the Sham and Trans group (P<0.05). Moreover, immunofluorescence assay also showed that LC-3 protein mainly distributes along renal tubular epithelial cells in a linear manner. CONCLUSIONS Autophagy dysfunction and inflammatory response after renal transplantation play important roles in processes of IRI. HBO treatment protects against the renal injury of IRI in renal tissues at the early stage, which may be triggered by the IL-6 pathway.

Pretreatment thrombocytosis as a prognostic factor in women with gynecologic malignancies: a meta-analysis.

BACKGROUND: This study was performed to analyze the prognostic implications of pretreatment or preoperative thrombocytosis in women with gynecologic malignancies. MATERIAL AND METHODS: We surveyed 2 medical databases, PubMed and EMBASE, to identified all relevant studies. A total of 14 (n=3,490) that evaluated the link between thrombocytosis and 5-year survival were included. REVMAN version 5.1 was used for our analysis and publication bias was evaluated using the Begg's funnel plot and tested by STATA 11.0. Risk ratios (RRs) with 95% confidence intervals (CIs) generated by the random effect model were used to assess the strength of any association. RESULTS: 709(20.3%) of the 3,490 patients exhibited thrombocytosis (platelet counts >400?109/L) at primary diagnosis, and their mortality was 1.62-fold higher compared with the others (RR=1.62, 95%CI= [1.28- 2.05], p<0.0001). Thrombocytosis failed to have a stronger effect on the survival of advanced patients of stages III to IV in our study (n=478, RR=1.29, 95% CI= [1.13-1.48], p=0.0003), nor in women with cervical cancer in stage IB (n=1371, RR= 1.73, 95% CI= [1.71-2.58], p=0.007). In addition, when adjusted for different carcinoma, it was associated with worse prognosis for all except the ones with vulvar cancer (n=201, RR= 0.43, 95% CI= [0.14-1.29], p=0.13). CONCLUSIONS: This meta-analysis indicated that thrombocytosis might be associated with a worse prognosis for patients with gynecologic malignancies but without specificity or sensitivity for the ones in advanced stage. When adjusted for different gynecologic malignancies, it showed a significant effect on survival of all except vulvar cancers.