Co-circulation of Hantavirus, Pathogenic Leptospira spp., and Bartonella spp. in Rodents in the Wanzhou Section of the Three Gorges Reservoir Region, 2021-2023.

Background: Rodent is a reservoir of various zoonotic pathogens. Wanzhou section of the Three Gorges reservoir region (TGRR) is a superior habitat for rodents, and the situation of rodent-borne zoonotic pathogens in this region has not been surveyed in recent years. Materials and Methods: Rodents were night trapped with mousecage or mousetrap in urban and surrounding towns' indoor or outdoor areas of the Wanzhou section of the TGRR, and nucleic acid was extracted from their lung or a mixture of liver, spleen, and kidney. Commercialized qPCR kits for pathogenic Leptospira spp., Rickettsia typhi, Anaplasma phagocytophilum, Bartonella spp., Orientia tsutsugamushi, and Francisella tularensis and qRT-PCR kits for hantavirus (HV), and severe fever with thrombocytopenia syndrome virus (SFTSV) were used for the detection of associated pathogens in collected rodents. Results: From 2021 to 2023, 604 rodents belonging to 10 species were collected. HV and pathogenic L. spp. were detected positive, with infection rates of 0.66% (4/604) and 1.32% (8/604), respectively. B. spp. were detected positive with an infection rate of 4.73% (19/402) in the rodents trapped in 2022 and 2023. Other five pathogens were all detected negative. Conclusion: This study showed that the Wanzhou section of the TGRR had HV, pathogenic L. spp., and B. spp. co-circulation in rodents. Hence, more attention should be paid to the prevention and control of associated rodent-borne diseases.

Effectiveness of predicting the spatial distributions of target contaminants of a coking plant based on their related pollutants.

The prediction accuracy of the spatial distribution of soil pollutants at a site is relatively low. Related pollutants can be used as auxiliary variables to improve the prediction accuracy. However, little relevant research has been conducted on site soil pollution. To analyze the prediction accuracy of target pollutants combined with auxiliary pollutants, Cu, toluene, and phenanthrene were selected as the target pollutants for this study. Based on geostatistical analysis and spatial analysis, the following results were obtained. (1) The reduction in the root mean square errors (RMSEs) for Cu, toluene, and phenanthrene with multivariable cokriging was 68.4%, 81.6%, and 81.2%, respectively, which are proportional to the correlation coefficient of the relationship between the auxiliary pollutants and the target pollutants. (2) The RMSEs calculated for the multivariable cokriging were lower than those obtained by only combining one related pollutants, and two co-variables should be better. (3) The predicted results for Cu, phenanthrene, and toluene and their corresponding related pollutants are more accurate than the results obtained not using the related pollutants. (4) In the interpolation process, the RMSEs for Cu, toluene, and phenanthrene with multivariable cokriging basically increase as the neighborhood sample data increases, and then they become stable. (5) When 84, 61, and 34 sample points were removed, the RMSEs for Cu, toluene, and phenanthrene, respectively, with multivariable cokriging were close to the RMSEs of the target pollutants based on the total samples. The results are of great significance to improving the prediction accuracy of the spatial distribution of soil pollutants at coking plant sites.

Identification of small extracellular vesicle subtypes in follicular fluid: Insights into the function and miRNA profiles.

The study of small extracellular vesicles (sEVs) heterogeneity is one of the main problems that must be solved, and the different sEV subtypes in follicular fluid are still unclear, limiting our understanding of their function. This study first separated sEV subtypes from follicular fluid using differential ultracentrifugation combined with iodixanol density gradient flotation and then evaluated their miRNA profile and effects on the proliferation and apoptosis of granulosa cells (GCs). We also performed Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of potential target genes of differentially expressed miRNAs (DEMs) and KEGG analysis of potential target genes of non-DEMs. Low-density sEVs (sEV_F6) were enriched in TSG101, while high-density sEVs (sEV_F8) were enriched in CD63. The miRNA profiles of sEV_F6 and sEV_F8 were heterogeneous, and the differential signaling pathways were mainly related to the adhesion and hypoxic stress pathways, while the same signaling pathways were mainly related to cell proliferation, apoptosis, cell cycle, and autophagy pathways. In addition, the highly expressed miRNAs in both subtypes were mainly related to cell proliferation and apoptosis. Both subtypes transferred their miRNAs into GCs and promoted the proliferation ability of the GCs and inhibited their apoptosis. The results showed for the first time that there are different subtypes of sEVs in follicular fluid and that the miRNA profiles of subtypes are heterogeneous.

[Effect of extracellular vesicles and microRNAs in follicular fluid on follicular development].

Extracellular vesicles (EVs) refer to bilayer membrane transport vesicles secreted by cells. EVs can take macromolecules from cells and transfer them to receptor cells. Among these macromolecular substances, the most studied are microRNAs (miRNAs). miRNA is non-coding RNA involved in the regulation of gene expression. It has been confirmed that there are different non-coding RNAs in mammalian follicular fluid EVs. EVs carrying miRNA can act as an alternative mechanism for autocrine and paracrine, affecting follicular development. This paper systematically introduced the kinds, characteristics and methods of isolation and identification of EVs, focusing on the effects of EVs and miRNAs on follicular development, including early follicular development, oocyte maturation, follicular dominance and effects on granulosa cell function. At the same time, the authors prospected the future research of EVs and microRNAs in follicular fluid, and provided ideas and directions for the research and application of EVs and miRNA functions in follicular fluid.

Evidence that downregulation of Wilms' tumor 1 (WT1) is involved in cortical stromal cell differentiation into theca cells in adult bovine ovaries.

Bovine theca cells are thought to differentiate from cortical stromal cells, and ovary-derived Wilms' tumor 1+ (WT1+ ) cells are the primary source of mouse theca cells. However, it is not known whether the differentiation of cortical stromal cells is regulated by WT1. Here, we identified WT1 in the cortical stroma and theca layer of the bovine ovary and analyzed the theca cell functional markers in cortical stromal cells and theca cells; in addition, we determined the effects of this gene on the secretion of androstenedione and progesterone by cortical stromal cells and the responsiveness of cortical stromal cells to luteinizing hormone (LH) in vitro. We used quantitative reverse-transcription polymerase chain reaction (RT-qPCR), western blot analysis, and immunohistochemistry to discover that the cortical stroma had higher WT1 expression than the theca layer. We used RT-qPCR and ELISA analyses to determine that the cortical stromal cells had lower levels of androstenedione and progesterone secretion and LHR messenger RNA expression than the levels of the theca cells. In cultured bovine cortical stromal cells, we found that WT1 downregulation increased androstenedione and progesterone secretion but had no effect on the LH responsiveness. Notably, the increase in androstenedione and progesterone secretion was associated with an increase in 3-ß-hydroxysteroid dehydrogenase expression. In conclusion, the results suggest that WT1 is involved in the differentiation of cortical stromal cells into cells with characteristics similar to theca cells of antral follicles in adult bovine ovaries.

Melatonin Stimulates STAR Expression and Progesterone Production via Activation of the PI3K/AKT Pathway in Bovine Theca Cells.

Melatonin is present in mammalian follicular fluid and plays an important role in regulating steroidogenesis in follicular cells. In this study, we report the effect of melatonin on steroidogenesis in the theca interna (TI) in small bovine follicles and theca cells (TCs) cultured in vitro. Treatment with melatonin significantly increased the expression of steroidogenic acute regulatory protein (STAR) and the production of progesterone in both TI and in TCs. Melatonin stimulated the phosphorylation of AKT but not ERK1/2, and the addition of luzindole (a nonspecific MT1 and MT2 inhibitor) or 4P-PDOT (specific MT2 inhibitor) reduced melatonin-induced STAR expression, progesterone secretion, and PI3K/AKT pathway activation. The effect of melatonin on the TI in follicles was more obvious than on the TCs in vitro. Results indicate that melatonin stimulates the steroidogenesis of TCs mainly via the activation of the PI3K/AKT pathway by MT1 and MT2.

The Wilms tumor gene (WT1) (+/-KTS) isoforms regulate steroidogenesis by modulating the PI3K/AKT and ERK1/2 pathways in bovine granulosa cells†.

The Wilms tumor (WT) gene WT1 encodes the splicing variants WT1(+KTS) and WT1(-KTS). Recent data suggest that WT1 plays an important role in the development of mice follicles. However, the mechanism through which WT1 influences ovarian steroidogenesis remains unknown. This study identified WT1 and evaluated the impact of splicing variants WT1(+KTS) and WT1(-KTS) on steroidogenesis using adult bovine granulosa cells (GCs). Using RT-qPCR and western blotting, we found that the ratio between WT1(+KTS) and WT1(-KTS) was stabilized. WT1 expression, however, decreased gradually in bovine GCs in response to follicle enlargement or atresia. The downregulation of WT1 increased the secretion of basal and follicle-stimulating-hormone-induced progesterone (P4), but decreased the secretion of basal-induced estradiol (E2). This was associated with an increase in the expression of 3ß-HSD, and a decrease in the expression of CYP19A1. In addition, WT1(-KTS) overexpression suppresses the secretion of E2 and P4 compared with WT1(+KTS) overexpression. This was associated with a decrease in the expression of CYP19A1, CYP11A1, and 3ß-HSD in cultured bovine GCs. Of note, the downregulation of WT1 suppresses the phosphorylation levels of AKT and p-ERK1/2. However, WT1(-KTS) overexpression promotes the phosphorylation levels of AKT and suppresses p-ERK1/2 levels. LY294002 (AKT inhibitor) increases MKP3 mRNA expression levels but decreases the level of p-AKT and p-ERK1/2. Collectively, WT1 significantly suppresses the mRNA expression of CYP11A1 and 3ß-HSD and the secretion of P4 in bovine GCs. Moreover, it regulates CYP19A1 mRNA expression and E2 secretion with complex networks, at least in part, by modulating AKT and ERK1/2 signaling. The effect of WT1(-KTS) was more pronounced than that exerted by WT1(+KTS).