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Previous studies have shown that increasing estradiol concentrations had a toxic effect on the embryo and were deleterious to embryo adhesion. In this study, we evaluated the physiological impact of estradiol concentrations on endometrial cells to reveal that serum estradiol levels probably targeted the endometrium in controlled ovarian hyperstimulation (COH) protocols. An attachment model of human choriocarcinoma (JAr) cell spheroids to receptive-phase endometrial epithelial cells and Ishikawa cells treated with different estradiol (10−9 M or 10−7 M) concentrations was developed. Differentially expressed protein profiling of the Ishikawa cells was performed by proteomic analysis. Estradiol at 10−7 M demonstrated a high attachment rate of JAr spheroids to the endometrial cell monolayers. Using iTRAQ coupled with LC–MS/MS, we identified 45 differentially expressed proteins containing 43 significantly upregulated and 2 downregulated proteins in Ishikawa cells treated with 10−7 M estradiol. Differential expression of C3, plasminogen and kininogen-1 by Western blot confirmed the proteomic results. C3, plasminogen and kininogen-1 localization in human receptive endometrial luminal epithelium highlighted the key proteins as possible targets for endometrial receptivity and interception. Ingenuity pathway analysis of differentially expressed proteins exhibited a variety of signaling pathways, including LXR/RXR activation pathway and acute-phase response signaling and upstream regulators (TNF, IL6, Hmgn3 and miR-140-3p) associated with endometrial receptivity. The observed estrogenic effect on differential proteome dynamics in Ishikawa cells indicates that the human endometrium is the probable target for serum estradiol levels in COH cycles. The findings are also important for future functional studies with the identified proteins that may influence embryo implantation.

Self-ordering Anodic Aluminum Oxide Films Using Multi-step Anodizing MA Di,, LI Shu-bai, DONG Xin-yan HU Xiu-ying, WANG Bin, CHEN Xiang-yan (1School of Chem. & Environ. Engin., Jiangsu Univ. of Tech., Changzhou, Jiangsu, 213001 E-mail: amandama2008@gmail.com) (2Dept. of Chem. Engin., Changzhou Inst. of Engin. Tech., Changzhou 213164) (3 School of Biotech. & Environ. Engin., Ningbo Inst. of Tech., Zhejiang University, Ningbo 315100) Anodic aluminum oxide(AAO) film is known to have highly ordered pore arrangement, controllable pore diameter and well-characterized morphology when produced in acidic electrolytes. As a self-organized fine structure with a nanohole array, porous AAO has been used as a host material for many applications. Nevertheless, a full recognition of the potential of AAO a template for nano-fabrication only developed in the last few years since the pioneering work of Masuda’s group. A variety of nanostructures can be created using AAO templates by depositing metals, semiconductors, organics, or their combinations into the pores. For example, metallic nanowires and hollow nanotube arrays, were synthesized by controlled electrodeposition of a metal salt in an AAO template. In the present work, we studied the self-ordering process of the AAO films during multi-step anodization. During the leaching process, the porous alumina film was dipped in phosphoric acid and chromic acid solution for etching film. Each anodization process was followed by this leaching process.

The surface of porous anodic aluminum oxide (AAO) film anodizing in malonic acid, which is characterized by Scanning Electron Microscope (SEM) and ImageJ software. There are disorderly tiny pores or stripes on the first once anodizing surface. Pore diameter, pore density and porosity are decided by the first anodizing process. With anodizing step increased, pore diameter of the membrane decreased. Two-step anodization improves the order of PAA membrane greatly, which is processed on the basic of the ordered array pits at the aluminum that is observed after removing membrane of the one-step anodization. According to the experiments, porous anodic aluminum oxide (PAA) was prepared in 1.0 mol/L malonic acid, its pore diameter increased and porosity decreased with anodizing voltage increased.

Accumulating evidence suggests a role of bisphenol A (BPA) in metabolic disorders. However, the underlying mechanism is still unclear. Using a mouse BPA exposure model, we investigated the effects of long-term BPA exposure on lipid metabolism and the underlying mechanisms. The male mice exposed to BPA (0.5 μg BPA /kg/day, a human relevant dose) for 10 months exhibited significant hepatic accumulation of triglycerides and cholesterol. The liver cells from the BPA-exposed mice showed significantly increased expression levels of the genes related to lipid synthesis. These liver cells showed decreased DNA methylation levels of Srebf1 and Srebf2, and increased expression levels of Srebf1 and Srebf2 that may upregulate the genes related to lipid synthesis. The expression levels of DNA methyltransferases were decreased in BPA-exposed mouse liver. Hepa1-6 cell line treated with BPA showed decreased expression levels of DNA methyltransferases and increased expression levels of genes involved in lipid synthesis. DNA methyltransferase knockdown in Hepa1-6 led to hypo-methylation and increased expression levels of genes involved in lipid synthesis. Our results suggest that long-term BPA exposure could induce hepatic lipid accumulation, which may be due to the epigenetic reprogramming of the genes involved in lipid metabolism, such as the alterations of DNA methylation patterns.

Valone has a chronic and toxic anticoagulant rodenticide that has widely used in China and has resulted in some accidental and intentional intoxication in recent years. The literature reported so far lacks sensitive and selective method for the confirmation of valone. The purpose of this study was to establish a novel assay for the identification and quantification of valone in serum by ion chromatography–electrospray ionization tandem mass spectrometry (IC–MS/MS). After serum sample was extracted with methanol/acetonitrile (10:90, v/v) and cleaned by Oasis ® HLB solid-phase extraction cartridge, chromatographic separation was performed on an Ionpac ® AS11 column with an eluent of methanol/30 mmol/L KOH (10:90, v/v). The overall extraction efficiency was >81.0%, and the limit of quantification was 0.5 ng/mL for valone. Regression analysis of the calibration data revealed good correlation ( r 2 > 0.99) for valone. Intra- and inter-day precisions for quality-control samples were less than 8.0 and 13.7%, respectively. The proposed method enables the identification and quantification of valone in both clinical and forensic specimens.