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Hydrogen sulfide (H2S), a gaseous intracellular signal transducer, participates in multiple physiological and pathological conditions, including reproductive conditions, and disrupts spermatogenesis. The blood-testis barrier (BTB) plays a vital role in spermatogenesis. However, the effect of H2S on the BTB and the underlying mechanism remain unclear. Herein, we examined the effect of H2S and omega-3 polyunsaturated fatty acids (ω-3 PUFAs) on the BTB and testicular functions. ICR male mice were randomly divided into the following groups: control, H2S exposure, and H2S exposure with ω-3 PUFAs intervention. The sperm parameters (sperm concentration and sperm motility) declined in the H2S group and improved in the ω-3 intervention group. BTB integrity was severely disrupted by H2S, and the BTB-related gene levels (ZO-1, Occludin, Claudin 11) decreased; ω-3 supplementation could alleviate BTB disruption by upregulating BTB-related genes, and TM4 Sertoli cells had a similar trend in vitro. p38 MAPK phosphorylation was upregulated in the Na2S treatment group and downregulated after ω-3 cotreatment. These findings suggest that H2S can impair the BTB and that ω-3 PUFAs supplementation can attenuate H2S toxicity in the male reproductive system. Our study elucidated the relationship between a gasotransmitter (H2S) and the BTB and identified the potential therapeutic effect of ω-3 PUFAs.

Exosomes are a type of extracellular vesicles derived from cells and mediators of intercellular communication. Different cell types have their own unique exosomes for exchanging information. We previously found that SASH1, a tumor suppressor, was lowly expressed or absent in glioma tissues and glioma C6 cells, but the structure and function of the corresponding exosomes had been unclear. Hence, we aimed to investigate whether exosomes generated from normal glial cells and glioma cells form different protein patterns and whether those derived from normal glial cells affect SASH1 expression in glioma cells. We collected exosomes from astrocytes and C6 cells and identified their exosomal proteins through mass spectrometry. We also performed gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analyses, whose results showed that both the total and unique exosomal proteins from each cell type were similar. Moreover, the KEGG analysis revealed different clusters of unique exosomal proteins in glial cells and glioma cells. In the normal glial cells, the top clusters were mainly involved in processes with RNA transcripts and proteins, whereas in glioma cells the clusters were attributed to PI3K-Akt signaling, cell adhesion, and cancer-related pathways. Western blot analysis showed that HMGB1 exists in exosomes derived from cultured astrocytes, although its expression was higher in glioma C6 cells. Furthermore, we found that exosomes extracted from astrocytes could increase SASH1 expression in C6 cells (P = 0.040), whereas those derived from HMGB1-depleted astrocytes could not (P = 0.6133). The expression levels of SASH1 decreased after the addition of extracellular recombinant HMGB1 protein, whereas that of TLR4 increased. Our study is the first to demonstrate that HMGB1 plays different roles depending on its form: as an extracellular protein, HMGB1 decreases SASH1 expression, but as an exosomal protein, HMGB1 increases SASH1 expression. Nevertheless, the mechanism, which partly depends on the TLR4 pathway, behind these opposing effects requires further study. Our novel findings on the structure-dependent roles of the cytokine HMGB1 in promoting or inhibiting cancer provide a fresh insight into the interactions of cancer cells with the microenvironment.

Circular RNAs (circRNAs) are important for the development and regeneration of the nervous system. We investigated the differential expression profiles of circRNA induced by spinal cord injury and reported that circRNA_01477 facilitates spinal astrocyte proliferation and migration after injury in rats. In this study, we further clarified the function and possible mechanism of action of circRNA_01477 in neurons. Fluorescence in situ hybridization assay revealed that circRNA_01477 is mainly localized in the neuronal cytoplasm. Knockdown of circRNA_01477 significantly increased axonal length. The circRNA_01477/microRNAs (miRNA)/messenger RNA (mRNA) interaction network was investigated using RNA sequencing. miRNA-3075 showed a remarkable increase after circRNA_01477 depletion, and either overexpression of miRNA-3075 or downregulation of its target gene FosB significantly promoted axonal growth. Luciferase reporter assay showed that miRNA-3075 could directly bind to the 3'UTR of FosB and negatively regulated FosB transcription. Dual silencing of circRNA_01477 and miR-3075 revealed that miR-3075 inhibition rescued the increased axon length caused by siCircRNA_01477. Finally, we verified that the Stat3 pathway was activated after FosB protein depletion in rat spinal neurons, while the NF-κB pathway was not altered. In summary, our study is the first to report that circRNA_01477 contributes to axon growth by functioning as miRNA sponge by regulating the miRNA-3075/FosB/Stat3 axis.

A metamaterial equipment is designed and experimental verified in the near-infrared with two reflectivity dips. The metamaterial equipment shows independent of polarization. Simulated results indicate that the reflectivity dip is excited by the coupling of localized surface plasmon (LSP) modes. The metamaterial equipment can work as a refractive index detection sensor with high figure of merit (FOM) value. This proposed metamaterial sensor can be applied in detecting different biochemical liquid.

In this paper, a high performance metamaterial absorber is designed and experimental studied. Measured results indicate that a perfect absorption band and a short-wavelength absorption peak are achieved in the near-infrared spectrum. Current strength distributions reveal that the absorption band is excited by the cavity resonance. And electric field distributions show that the short-wavelength absorption peak is excited by the horizontal coupled of localized surface plasmon (LSP) modes near hole edges. On the one hand, the absorption property of the measured metamaterial absorber can be enhanced through optimizing the structural parameters ( a, w , and H ). On the other hand, the absorption property is sensitive to the change of refractive index of environmental medias. A sensing scheme is proposed for refractive index detecting based on the figure of merit (FOM) value. Measured results indicate that the proposed sensing scheme can achieve high FOM value with different environmental medias (water, glucose solution).

Operation energy consumption of heat pump systems is one of the key problems for system design. This study establishes the models of transportation energy consumption ratio and transportation distance limit and puts forward the concepts of new economic specific frictional resistance. According to existing calculation formulas of specific frictional resistance, calculation formulas for more economic specific frictional resistance are deduced by combining with economic indexes. In this way, a range (30–400 Pa/m) of new economic specific frictional resistance is obtained to determine the transportation energy consumption ratio of water. On this basis, the pipe network is designed to reduce and control the transportation energy consumption ratio of pumps to 15%-20%.

This study aims to evaluate the effects of immersion vaccination through hyperosmotic treatment, and determine the optimum hyperosmotic salinity for largemouth bass (Micropterus salmoides) by determining immune responses and protective efficacies post vaccination. For hyperosmotic immersion (HI), M. salmoides were immersed in three hyperosmotic salinities (5, 10 and 20‰ salinity) respectively, then transferred into normal freshwater containing inactivated Aeromonas veronii. For direct immersion (DI) group, fish were directly immersed in freshwater with the same concentration of bacterin. Non-specific immune responses were evaluated based on the analysis of nonspecific humoral substances and immune related genes (interleukin 1β, CD40, IL-8 and tumor necrosis factor α), the results showed that serum lysozyme and myeloperoxidase activities were appreciably up-regulated earlier and maintained higher levels in HI groups than DI group, highest activities were observed in 10‰ salinity HI group. The expression dynamics of immune related genes had similar trend with nonspecific humoral substances. For specific cellular immune responses, the percentages of surface membrane immunoglobulin-positive (sIg+) cells in DI group did not show any significant increase until at 14 day post immunization (dpi) and peaked at 35 dpi (23.17%), sIg+ cells in HI groups showed a marked rise 7 days earlier than DI group, then steadily increased and peaked at 28 dpi (5‰ salinity: 25.83%; 10‰ salinity: 32.03%; 20‰ salinity: 24.17%). The serum specific antibody level showed obvious increase in all vaccination groups since 7 dpi, it reached the peak at 28 dpi in HI groups (5‰ salinity: OD = 0.49; 10‰ salinity:OD = 0.71; 20‰ salinity:OD = 0.50), one week earlier than DI group. Both sIg+ cells and specific antibody level were highest in the 10‰ salinity HI group. The relative percent survival (RPS) was 68.42%, 89.42% and 63.16% in 5, 10 and 20‰ salinity HI groups, respectively, higher than this in DI group (47.37%). These results demonstrated that HI at 10‰ salinity could efficiently enhance the immune responses of M. salmoides and provide higher RPS. This has significant value for hyperosmotic immersion with bacterin in M. salmoides.

Precise diagnosis and effective treatment of cancer metastasis remain a great challenge in the clinic. Theranostic agents that consist of both diagnostic and therapeutic components have gained great interest in cancer therapy. Herein, we report an intraoperative theranostic strategy for the accurate live detection of micrometastases with selective chemotherapeutic activity using a small multifunctional agent (CBHAc). Combining histone deacetylase (HDACs) targeting and pH-responsive fluorescence characteristics, CBHAc provided highly sensitive and selective real-time fluorescence diagnosis for microtumors in zebrafish and hepatic micrometastases in mice. Furthermore, CBHAc demonstrated significant tumor inhibition against colon cancer and hepatic matastasis, prolonging the survival of tumor-bearing mice through attenuating epithelial-mesenchymal transition (EMT). Therefore, this cancer-targeting fluorescence agent delivers a new platform for both intraoperative diagnosis and suppression of cancer metastasis in surgery, broadening the impact of multifunctional agents for theranostic precision medicine.

Cyprinid herpesvirus 2 (CyHV-2) is the causative agent of haematopoietic necrosis disease in farmed gibel carp (Carassius auratus gibelio). It appears to be spreading cross the world, and has resulted in massive mortality and great economic losses. Although many sensitive nucleic acid-based diagnostic methods have been developed, effective immunodiagnosis and neutralization approaches based on monoclonal antibodies (MAbs) against CyHV-2 membrane protein are still scarce to study. In this study, ORF25 gene was chemically synthesized and cloned into a pET-28a expression vector to produce rORF25 (recombinant ORF25) in E. coli BL21 (DE3). The purified rORF25 was served as an immunogen to prepare monoclonal antibodies (MAbs), and five efficient hybridoma cell lines were selected by enzyme-linked immunosorbent assay (ELISA). Among them, MAbs 7H10-1B6 and 1F8-1B6 belonged to the IgG2b isotype, while the other three MAbs designed as 8D9-1H6, 3H2-1G5 and 2C3-1E6 belonged to the IgG1 isotype. Western blotting results indicated that MAbs could specifically identify the rORF25. MAb 2C3-1E6 could specifically detect ORF25 in CyHV-2 infected Fathead Minnow cells and gible carp tissues by ELISA, indirect immunofluorescence and immunohistochemistry. In vitro neutralization assay showed that MAb 2C3-1E6 exhibited a titer-dependent neutralization effect on CyHV-2 infection in FHM cells. The results indicated that MAbs against rORF25 could serve as effective detection probes and potential neutralizing antibody for haematopoietic necrosis disease in gibel carp.

Lymphocystis disease virus (LCDV) enters and infects the gill cells of flounder (Paralichthys olivaceus) via a 27.8 kDa membrane protein receptor. In the present study, immunohistochemistry was performed to locate the tissue distribution of this molecule in healthy flounder and showed that it was widely distributed in the tissues tested. Indirect enzyme-linked immunosorbent assay (ELISA) showed that the expression of the receptor in healthy flounder was highest in the gills and stomach, then in the skin, intestine and liver, followed by the spleen, head kidney, heart, ovary and brain and finally the kidney. On LCDV infection, ELISA indicated that the expression of the receptor, as determined by ELISA, was significantly upregulated in all tissues of LCDV-infected flounder compared with controls, but this expression decreased over the 4 weeks post infection. In contrast, real-time quantitative polymerase chain reaction demonstrated that the copy number of the LCDV gene in the tissues increased with time post infection, and that viral loads were higher in the tissues with higher expressions of the receptor. These results point to a correlation between high expression of the 27.8 kDa receptor and efficient LCDV propagation. The wide tissue distribution of the receptor might be one reason why LCDV can infect various tissues leading to systemic infection.

Histone H3 lysine 27 acetylation (H3K27ac) is an active epigenetic modification which has been revealed to be associated with active gene expression. It was hypothesized that H3K27ac might also participate in the porcine somatic reprogramming process during early development of SCNT-derived embryos. The spatial and temporal expression profiles of H3K27ac were investigated at different developmental stages in SCNT embryos compared with in vitro fertilization (IVF) and parthenogenetic activation (PA) counterparts. Specifically, results showed that amounts of H3K27ac gradually decreased from the earliest pronuclear stage to 8-cell stage, corresponding to the major embryonic genome activation (EGA), followed by re-acetylation of H3K27 from the morula stage onwards accompanying the first cell lineage specification in IVF embryos. Similar dynamic patterns of H3K27ac signal was observed at all developmental stages of porcine SCNT and PA embryos except for the hatched stage in which amounts of H3K27ac in SCNT and PA embryos was slightly less than that in IVF counterparts. Moreover, the gradual decrease of H3K27ac before EGA was demonstrated to be an active process independent of DNA replication, RNA and protein synthesis. The expression of HDAC1, HDAC2, MBD3 and CBP genes were well correlated with the dynamic changes of H3K27ac mark. Overall, these results indicate that H3K27ac is only defective in late SCNT blastocysts, and that the dynamic changes of this marker might also underlie the EGA and initial cell lineage specification during early embryo development.

DNA active demethylation is an important epigenetic phenomenon observed in porcine zygotes, yet its molecular origins are unknown. Our results show that 5-methylcytosine (5mC) converts into 5-hydroxymethylcytosine (5hmC) during the first cell cycle in porcine in vivo fertilization (IVV), IVF, and SCNT embryos, but not in parthenogenetically activated embryos. Expression of Ten-Eleven Translocation 1 (TET1) correlates with this conversion. Expression of 5mC gradually decreases until the morula stage; it is only expressed in the inner cell mass, but not trophectoderm regions of IVV and IVF blastocysts. Expression of 5mC in SCNT embryos is ectopically distinct from that observed in IVV and IVF embryos. In addition, 5hmC expression was similar to that of 5mC in IVV cleavage-stage embryos. Expression of 5hmC remained constant in IVF and SCNT embryos, and was evenly distributed among the inner cell mass and trophectoderm regions derived from IVV, IVF, and SCNT blastocysts. Ten-Eleven Translocation 3 was highly expressed in two-cell embryos, whereas TET1 and TET2 were highly expressed in blastocysts. These data suggest that TET1-catalyzed 5hmC may be involved in active DNA demethylation in porcine early embryos. In addition, 5mC, but not 5hmC, participates in the initial cell lineage specification in porcine IVV and IVF blastocysts. Last, SCNT embryos show aberrant 5mC and 5hmC expression during early porcine embryonic development.

Human bactericidal/permeability-increasing protein (hBPI) is the only antibacterial peptide which acts against both gram-negative bacteria and neutralizes endotoxins in human polymorphonuclear neutrophils; therefore, hBPI is of great value in clinical applications. In the study, we constructed a hBPI expression vector (pBC1-Loxp-Neo-Loxp-hBPI) containing the full-length hBPI coding sequence which could be specifically expressed in the mammary gland. To validate the function of the vector, in vitro cultured C127 (mouse mammary Carcinoma Cells) were transfected with the vector, and the transgenic cell clones were selected to express hBPI by hormone induction. The mRNA and protein expression of hBPI showed that the constructed vector was effective and suitable for future application in producing mammary gland bioreactor. Then, female and male goat fibroblasts were transfected with the vector, and two male and two female transgenic clonal cell lines were obtained. Using the transgenic cell lines as nuclear donors for somatic cell nuclear transfer, the reconstructed goat embryos produced from all four clones could develop to blastocysts in vitro. In conclusion, we constructed and validated an efficient mammary gland-specific hBPI expression vector, pBC1-Loxp-Neo-Loxp-hBPI, and transgenic hBPI goat embryos were successfully produced, laying foundations for future production of recombinant hBPI in goat mammary gland.

This paper is intended for studying the computational accuracy and the fitting problem of “uncracked stress” in pipe-shaped structure details using isoparametric boundary element method. At first, the influences of different kinds of discretion plan on computational accuracy are studied by considering the simplest pipe-shaped structure—a thick wall cylinder, then the “Uncracked stress” distribution of typical pipe-shaped structures—“—shaped” and “T-shaped” junctions is calculated. In order to improve the fitting accuracy of “uncracked stress”, more internal nodes are added to the local area while more interpolation points are added to boundary element where the local stress gradient is great. The study shows that the computational accuracy is satisfactory even a few boundary elements and internal nodes are used, when the eight—node isoparametric boundary element method is adopted.