Immunotoxicity and uterine transcriptome analysis of the effect of zearalenone (ZEA) in sows during the embryo attachment period.

Zearalenone is a mycotoxin and a pollutant that is commonly found in crops. Once ingested, ZEA can cause disturbances in the immune system and produce immunotoxicity. However, there is little research on the effect of ZEA exposure on the relationship between immune regulation and embryo implantation in the uteri of sows. Embryo implantation relies upon the fact that the relationship between the maternal and fetal immune systems is balanced. This balance is provided by the joint regulation of immune organs, cytokines, and uterine immunity. In this study, we investigated 20 sows with an initial weight of 100.00 ± 5.00 kg and 200 days in age. The sows were fed with diets containing ZEA at concentrations of 0 mg/kg, 1 mg/kg, 2 mg/kg, and 10 mg/kg, respectively, from 8 to 14 days of gestation. We studied immunotoxicity and the uterine transcriptomics associated with the effect of ZEA in sows during embryo attachment. Following ZEA treatment, serum biochemical analysis and RT-qPCR were used to detect the concentration and mRNA expression levels of immunoglobulin IgA, IgG, and IgM, in the serum and spleen, respectively. The same analysis was carried out for a range of cytokines in the serum and spleen: IL-1, IL-2, IL-6, IL-10, and TNF. Uterine transcriptome analysis revealed 75, 215, and 81 genes that were differentially expressed in the 0 mg/kg vs 1 mg/kg treatment, 0 mg/kg vs 10 mg/kg treatment, and 1 mg/kg vs 10 mg/kg treatment, respectively. GO terms analysis showed that the up-regulated genes related to the immune system were highly expressed. KEGG pathway analysis further revealed the importance of several metabolic pathways, including drug metabolism-cytochrome P450, the cytokine-cytokine receptor interaction pathway, and calcium signaling pathways. The differentially expressed genes were confirmed by quantitative real-time PCR. These findings expand our understanding of the gene expression profiles and signaling pathways associated with the immune response to ZEA exposure in sows during the embryo implantation window. This study provides valuable information for clarifying the molecular mechanism of ZEA's immunotoxicity to early pregnant sows in the future.

Zearalenone Blocks Autophagy Flow and Induces Cell Apoptosis During Embryo Implantation in Gilts.

Zearalenone (ZEA) has been proved to be toxic, particularly to the reproductive system of gilts. The effect of ZEA on gilts during embryo implantation window period is of particular interests. Here, we observed window stage dysontogenesis of gilts treated with ZEA. In endometrial tissues and cells, autophagosomes increased significantly and mitochondria were damaged with increasing ZEA concentration. Addition of autophagy inhibitor confirmed that ZEA blocks the autophagic flow in the fusion of autophagosomes and lysosomes. In conclusion, ZEA exposure during embryo implantation results in endometrium inflammation by activating autophagy while blocking autophagy flow at the same time, leading to the significant accumulation of autophagosomes. The aforementioned effects of ZEA induce the apoptosis of primary endometrial cells through the caspase3 pathway, which would break the uterus environment balance and finally lead to embryo implantation failure and dysontogenesis in gilts.

mRNA/lncRNA expression patterns and the function of fibrinogen-like protein 2 in Meishan pig endometrium during the preimplantation phases.

Embryonic implantation involves a complex and well-coordinated interaction between the developing conceptus and maternal uterus, and the preimplantation period has a major impact on litter size in pigs. The present study aimed to investigate the vital messenger RNAs (mRNAs) and long noncoding RNAs (lncRNAs) that regulate preimplantation in Meishan pigs. The enriched Gene Ontology terms were all related to "binding." Furthermore, "ECM-receptor interaction" was predicted as an important pathway that regulated the success of implantation. We speculated that the differentially expressed mRNAs S100A9, ANXA8, MUC16, and FGL2 and the differentially expressed lncRNAs TCONS_11206566, TCONS_09904861, and TCONS_1252933 may play vital roles in the process of implantation. Furthermore, this study verified that FGL2 was highly expressed on Day 12 of pregnancy, and we also investigated the function of FGL2 during preimplantation in vivo. In conclusion, this study provides useful information for further analyses of the molecular mechanisms of implantation in Chinese domestic pigs.

Degradation of methylene blue by dielectric barrier discharge plasma coupled with activated carbon supported on polyurethane foam.

The degradation of methylene blue (MB) using a novel dielectric barrier discharge plasma reactor coupled with activated carbon supported polyurethane foam (AC/PUF) was investigated in this paper. The plasma reactor combining a glass bead-packed bed and a microporous plate was developed. The AC/PUF provided sufficient contact area between carbon media and pollutants and hence revealed a good MB removal capacity. The effects of input voltage and initial MB solution concentration on MB degradation efficiency were examined. Kinetic study indicated that plasma and AC/PUF in the coupled system had a good synergistic effect in MB degradation. The degradation efficiency of 100 ppm MB solution could reach 97.9% with 10 min treatment in the coupled system, which was close to that obtained by plasma treatment alone for 30 min (97.5%). The COD removal in the plasma and AC/PUF coupled system (90.7%) was much higher than that obtained by plasma treatment followed by AC/PUF adsorption (58.3%). In addition, the energy yield (G 50) of the coupled system was up to 38.3 g kW-1 h-1, suggesting great energy efficiency of the system. Moreover, repeated use experiments of AC/PUF showed the good utilization potential of the coupled system. Finally, a possible degradation pathway of MB was proposed.

Sequence analysis of microRNAs during pre-implantation between Meishan and Yorkshire pigs.

Embryonic implantation in sows is a coordinated interaction between the implantation-competent blastocyst and receptive uterus. In addition, microRNAs are small endogenous non-coding RNAs which are involved in post-transcriptional gene regulation of several biological processes including embryonic implantation. However, the mechanisms of miRNAs involved in embryonic implantation of sows remain largely unknown. Here, we analyzed miRNAome of endometrium on day 9, 12 and 15 of pregnancy and on day 12 of non-pregnancy in Meishan and Yorkshire pigs by Illumina sequencing. From 24 libraries, we identified 312 known microRNAs and 211 potential novel miRNAs. Bioinformatics analysis showed that differentially expressed microRNAs on day 12 of pregnancy between the two breeds may play critical roles by involving "p53 signaling pathway" and "Wnt signaling pathway". Furthermore, our results demonstrated that ssc-miR-21, ssc-miR-451, ssc-miR-204, ssc-miR-199a-5p and ssc-miR-199b-5p would play crucial roles for implantation. The data generated in this study were expected to elucidate the influence of microRNAs during pre-implantation in pigs.

Identification of non-coding and coding RNAs in porcine endometrium.

One of the most critical periods of embryonic loss in pig is day 12 of pregnancy, when implantation begins. Here, we analyzed the gene expression on day 12 of pregnancy and non-pregnancy in the porcine endometrium using RNA sequencing (RNA-seq). 237 mRNAs, 34 lncRNAs and 1 miRNA were significantly differentially expressed between the two groups. Further functional analyses were conducted to identify these differentially expressed transcripts. The results demonstrated that they participate in various biological processes, such as cell adhesion, binding, nucleic and metabolic processes. In addition, our results showed that the differentially expressed genes (IL1R, FGF9, DUPS10, DUPS4, CD14 and MAP4K4) in MAPK pathway, and lncRNAs of XLOC_2604764 and XLOC_2604756 may play a vital role in regulating embryo implantation. Besides, we investigated the lncRNA-ssc-miR-132-mRNA interactions, aiming to explain the regulatory networks of coding and non-coding genes that contributes to the establishment of the maternal pregnancy.

Determination of protein by resonance light scattering technique using dithiothreitol-sodium dodecylbenzene sulphonate as probe.

The resonance light scattering (RLS) spectra of bovine serum albumin (BSA)-dithiothreitol (DTT)-sodium dodecylbenzene sulphonate (SDBS) and its analytical application were investigated. The RLS intensity of this system can be effectively enhanced in the presence of BSA. Based on the enhanced RLS intensity, a simple assay for BSA was developed. The experimental results indicate that the enhanced RLS intensity is proportional to the concentration of BSA in the range from 1.0 x 10(-8) to 7.5 x 10(-7) mol L(-1) with the determination limit of 5.0 x 10(-9) mol L(-1). The effects of pH, concentration of SDBS and DTT on the RLS enhancement were discussed. Most metal ions have little interference on the determination of BSA. Some synthetic and real samples were analyzed, and the results obtained were in good agreement with those obtained by Bradford method.