Objective To study the amount of rhizospheric microorganisms and soil enzyme activity influenced by Paridis Rhizoma in different locations and of different strains. Methods The amount of rhizospheric microorganisms, soil enzyme activity and their correlation were researched through field survey and collection of rhizospheric soil in Paridis Rhizoma cultivated in Three Gorges Reservoir Region and by microbial dilution plate culture method. Results The amount of rhizospheric microorganisms in Paridis Rhizoma from different habitats showed significant differences. The dominant species in soil microflora was bacteria; the second one was actinomycetes; the fewest one was fungus. The variation trend of the amount of rhizospheric microorganisms was not consistent with the variation trend of rhizospheric microorganisms diversity index. The activity of soil phosphatase, invertase and pepsin in Paridis Rhizoma from different habitats varied. The correlation analysis showed that the correlation between the soil enzyme activity and the amount of rhizospheric microorganisms existed. Conclusion Choosing the suitable strains and habitats of Paridis Rhizoma is beneficial to enhancing the amount of rhizospheric microorganisms and soil enzyme activity, which can create good micro-ecological environment for growth and cultivation of Paridis Rhizoma.

This study was aimed to examine the expression of apoptosis-associated gene Fas in HeLa cell, explore the effects of the co-immobilized cytokines (tumor necrosis factor-alpha and interferon-gamma), and probe the potential mechanism of action. The preparation and application of the research couple IFN-gamma and TNF-alpha to the polystyrene cell culture plate were performed using the Photo-immobilization method, with different doses (20 ng/well and 200 ng/well) and synthesized optical active material. HeLa cells were treated with cytokines for two dose and 1, 3, 6 days. The result showed that the free cytokines induced HeLa apoptosis quickly, yet the HeLa apoptosis induced by co-immobilized cytokines had longer effect.
Apoptosis ; drug effects ; genetics ; Drug Synergism ; HeLa Cells ; Humans ; Immobilized Proteins ; chemistry ; pharmacology ; Interferon-gamma ; chemistry ; pharmacology ; Tumor Necrosis Factor-alpha ; chemistry ; pharmacology ; Up-Regulation ; fas Receptor ; metabolism

BACKGROUND: The proliferation of peripheral blood stem cells among peripheral blood mononuclear cells (PBMCs) invitro remains unclear. There is no optimal marker for tracing PBMCs transplanted in vivo.OBJECTIVE: To observe the degree of PBMC proliferation in stem cell medium by EdU labeling and to explore thefeasibility of EdU-labeled peripheral blood stem cells.METHODS: New Zealand rabbit PBMCs were isolated and cultured for 1 to 5 days in stem cell medium supplementedwith EdU. The cells were observed and counted at 0, 1, 2, 3, 4 and 5 days in culture. The cells were harvested at eachtime point and stained with EdU fluorescent reagents. Then, confocal microscopy and flow cytometry were used to detectEdU-labeled cells.RESULTS AND CONCLUSION: (1) Freshly isolated rabbit PBMCs were rounded and showed clear outline. After 1 dayculture, most of the cells were suspended in the medium, spherical or round. There were also a few cell clusters andadherent cells scattered in a triangle or polygon shape; after 2 days culture, more cell debris were observed, and mostcells were round; when cultured for 3-5 days, increased cell debris, smaller cell mass and decreased cell densitysignificantly were observed. (2) With the prolongation of culture time, the cell count decreased gradually. (3) Whencultured for 1 day, EdU labeled cells in red were scattered. The number of cells marked with EdU red label increasedsignificantly at day 2 and remained unchanged after 3 days of culture. At 5 days of culture, the number of red cellsmarkedly decreased; the highest positive rate of EdU-labeled cells was (2.38±0.10)% at 2 days after culture. To conclude,these results showed that the proportion of proliferating cells in rabbit PBMCs was very low. EdU is capable of labelingproliferative cells among PBMCs.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide and macrophage polarization plays an important role in its pathogenesis. However, which molecule regulates macrophage polarization in NAFLD remains unclear. Herein, we showed NAFLD mice exhibited increased 17β-hydroxysteroid dehydrogenase type 7 (17β-HSD7) expression in hepatic macrophages concomitantly with elevated M1 polarization. Single-cell RNA sequencing on hepatic non-parenchymal cells isolated from wild-type littermates and macrophage-17β-HSD7 knockout mice fed with high fat diet (HFD) for 6 weeks revealed that lipid metabolism pathways were notably changed. Furthermore, 17β-HSD7 deficiency in macrophages attenuated HFD-induced hepatic steatosis, insulin resistance and liver injury. Mechanistically, 17β-HSD7 triggered NLRP3 inflammasome activation by increasing free cholesterol content, thereby promoting M1 polarization of macrophages and the secretion of pro-inflammatory cytokines. In addition, to help demonstrate that 17β-HSD7 is a potential drug target for NAFLD, fenretinide was screened out from an FDA-approved drug library based on its 17β-HSD7 dehydrogenase inhibitory activity. Fenretinide dose-dependently abrogated macrophage polarization and pro-inflammatory cytokines production, and subsequently inhibited fat deposition in hepatocytes co-cultured with macrophages. In conclusion, our findings suggest that blockade of 17β-HSD7 signaling by fenretinide would be a drug repurposing strategy for NAFLD treatment.