Azithromycin regulates Mettl3-mediated NF-κB pathway to enhance M2 polarization of RAW264.7 macrophages and attenuate LPS-triggered cytotoxicity of MLE-12 alveolar cells.

BACKGROUND: Azithromycin (AZM) has been proposed as a potential therapeutic drug in acute pulmonary injury due to its immunomodulatory and anti-inflammatory properties. However, its therapeutic mechanism remains not fully understood. METHODS: LPS was used to stimulate MLE-12 cells and RAW264.7 macrophages. Analyses of viability and apoptosis were performed by CCK-8 assay and flow cytometry, respectively. Protein analysis was performed by immunoblotting, and mRNA expression was tested by quantitative PCR. The secretion levels of TNF-α and IL-6 were detected by ELISA. MDA, GSH, ROS and Fe2+ contents were analyzed using assay kits. RESULTS: Administration of AZM or depletion of methyltransferase-like 3 (Mettl3) could attenuate LPS-triggered apoptosis, inflammation and ferroptosis in MLE-12 alveolar cells, as well as enhance M2 polarization of LPS-stimulated RAW264.7 macrophages. In LPS-exposed MLE-12 and RAW264.7 cells, AZM reduced Mettl3 protein expression and inactivated the NF-κB signaling through downregulation of Mettl3. Furthermore, Mettl3 restoration abated AZM-mediated anti-apoptosis, anti-inflammation and anti-ferroptosis effects in LPS-exposed MLE-12 cells and reversed AZM-mediated M2 polarization enhancement of LPS-exposed RAW264.7 macrophages. CONCLUSION: Our study indicates that AZM can promote M2 polarization of LPS-exposed RAW264.7 macrophages and attenuate LPS-triggered injury of MLE-12 alveolar cells by inactivating the Mettl3-mediated NF-κB pathway.

Urban park green space use analysis based on trajectory big data: Experience from a medium-sized city in China.

Regular visits to park green space offer remarkable benefits for the physical and mental health of urban residents. Achieving a comprehensive understanding of the utilization across the entire city is a prerequisite for improving the overall utilization rate of park green spaces. Traditional social survey methods are limited by their sample size and time-consuming nature, while methods based on geographic location big data are gaining momentum. This study focuses on Xuchang, a medium-sized city in China, and systematically analyzes the current state and influencing factors of park green space utilization by mining GPS trajectory big data from April 3 to 12, 2022. Results indicate that residents' choices of park green spaces are highly diverse. Approximately 20% of visitors on holidays and weekends, and about 25% of visitors on weekdays, prefer the park green space closest to their homes. Notably, the distance threshold for park green space visits on weekdays, weekends, and holidays is 3633, 3824, and 4127 m, respectively. These distances are significantly higher than the several hundred meters specified in planning documents or commonly used in accessibility analyses. For individuals who frequently visit park green spaces, distance is the most critical influencing factor. Conversely, for those who occasionally visit, distance is not the primary consideration. For individuals who rarely or never visit park green spaces, personal attitudes play an essential role. In comparison to weekdays, the number of visitors on holidays and weekends is larger, the travel distance is longer, and they are more inclined to choose larger parks. Visits are concentrated in the afternoon and evening, and weather changes remarkably affect park green space utilization. Importantly, no compensatory effect is observed between the frequency and duration of park green space visits. These findings hold important implications for urban planning, management, and the promotion of park green space utilization.

Network Pharmacology and Experimental Validation to Explore the Molecular Mechanisms of Compound Huangbai Liquid for the Treatment of Acne.

Background: Acne is a highly prevalent skin disease, and inflammation plays an important role. Compound Huangbai Liquid (CHL) is a classical traditional Chinese medicine (TCM) with remarkable clinical therapeutic effects on acne. However, a holistic network pharmacological approach to explain the mechanism of CHL in the treatment of acne has not been explored. Methods: In this study, active components and action targets of Compound Huangbai Liquid were assessed via BATMAN-TCM. The target genes related to acne were extracted from GeneCards, DisGeNet and OMIM databases. Venn diagrams to predict potential targets for the treatment of acne. Protein-Protein interaction (PPI) analysis was proceeded through String database to obtain the core protein, and the protein interaction network was constructed by Cytoscape 3.9.1. Gene Ontology (Go) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed on Metascape platforms and bioinformatics.com.cn. TCM-compound-target-disease network and disease-target pathway network were constructed using Cytoscape to give the visual result. Finally, the results were further verified by establishing the mouse acne animal model. Results: This approach identified 165 active compounds, 1117 gene targets, 156 acne-related targets, and 34 potential target proteins for the treatment of acne with CHL. The biological processes were primarily related to cellular response to lipid, response to lipopolysaccharide, and regulation of secretion. The CHL was significantly associated with ten pathways including the Chagas disease and pathways in cancer. Animal experiments showed that CHL could significantly alleviate the levels of inflammatory factors and TLR4/NF-κB/p38 MAPK signaling pathway in acne. Conclusion: This study revealed the multiple active components, multiple targets, and multiple pathways of CHL in the treatment of acne, which provided a new perspective for the study of the mechanism of traditional Chinese medicine in the treatment of acne.

Associations among ecosystem services from local perspectives.

Present works on the relationship between pairwise ecosystem services (ES) in one period are mostly based on global perspective, which ignores the influence of spatial heterogeneity. In view of this problem, this study proposes a static local association model for pairwise ES, and the applicability of this model is then tested in Ningxia Hui Autonomous Region, China. Four ES, namely, grain production, livestock production, soil conservation, and carbon sequestration, are selected. The global and local associations among these services are tested and compared using Spearman's correlation analysis and the proposed local static association model, respectively. Results showed that all six possible ES pairs were significantly correlated in the global scale. However, for strongly correlated pairwise ES (e.g., grain production and livestock production and carbon sequestration), a reverse trend still exists in some local areas. This inference is especially true for ES pairs with relatively low correlation coefficient. We also determine that some of the trade-offs or synergies are formed only by artifact, such as the synergy between grain and livestock production in the midwest, the trade-off between grain production and carbon sequestration concentrating on the southern mountains, and the trade-off between livestock production and soil conservation and carbon sequestration in the central area of the northern Yellow River irrigation district. In summary, local association analysis conveys the abstract overall relationship among specific spatial locations to identify the mechanism of the relationship among ES. This finding will provide new insights for the planning and management of ES, which include that local ES cannot be managed and planned according to the global relationship, local trade-offs or synergies caused by artifact cannot be applied to ES management, the proposed local association analysis helps identify ES problems in parts, and the high-value synergy area is an appropriate reference for ecosystem management.

Scale effect on spatial patterns of ecosystem services and associations among them in semi-arid area: A case study in Ningxia Hui Autonomous Region, China.

Identifying scale effect on spatial patterns of ecosystem services and associations among them has been recognized as critical to the sustainable management of ecosystem services. We proposed a method to conveniently integrate ecosystem services of small scale into different larger scales. Taking Ningxia Hui Autonomous Region in west China as an example, we analyzed the change regularity of spatial patterns of 7 ecosystem services under 22 different scales. Further, the tradeoffs and synergies among ecosystem services across all scales were compared. The results showed that all of the 7 ecosystem services had been characterized by stable spatial cluster patterns across all of the 22 scales in our study. However, the extent of aggregation decreased with the increase of scale owing to the 'peak cutting and valley filling' process of map scale synthesis. Most of the associations among ecosystem services were robust across scales. However, there was a trend that smaller scales had more pairwise correlations than larger scales. The formation of tradeoffs and synergies among ecosystem services can be attributed to one or more of the following three factors: land use conflict or consistency, common drivers, and interactions among ecosystem services. We attribute the change of relationships among ecosystem services with scales to that the role of factors causing tradeoffs and synergies among ecosystem services may change with scales. Food supply service synergizes with all of the 3 regulating services at almost all of the scales in our study area, indicating that well managed farmlands with high net profit are beneficial to soil conservation and organic matter accumulation in semi-arid area.

[Effects of Simvastatin on Expression of Multidrug Resistance Gene and Protein of SHI-1 Cells].

OBJECTIVE: To investigate the effects of simvastatin(SIM) and serum free medium(SFM) on the expression of multidrug resistance gene(MDR1) and protein of SHI-1 cells. METHODS: Trypan blue exclusion assay was used to detect the proliferation level and viability of SHI-1 cells after treatment with SIM and culture in SFM; The multi-drug resistant protein p-gp was measured by flow cytometry after culture in SFM for 1 to 3 days and treatment with various concentration of simvastatin. The effect of SFM culture and SIM treatment on the expression of MDR1 trascript was detected by qPCR; ELISA was used to measure the change of cellular cholesterol after culture in SIM and SFM. Chemosensitivity assay was performed after treatment with SIM for SHI-1 cells. RESULTS: Compared with control group, the growth of SHI-1 cells cultured in SFM decreased in a time-dependent manner. The growth-inhibitory effect was markedly increased when SHI-1 cells were treated with SIM and SFM. The mRNA level of MDR1 gene decreased after SIM treatment or/and culture in SFM. P-gp protein was downregulated in SHI-1 cells cultured in SFM or/and treated with SIM. The cellular cholesterol level increased when the cells were cultured in SFM. Total cellular cholesterol level decreased in SHI-1 cells treated with SIM and cultured in SFM. Chemosensitivity assays found that pre-treatment with SIM could increase the cytotoxicity of DNR to SHI-1 cells. CONCLUSION: Culture with SIM and SFM can downregulate the expression of MDR1 gene and p-gp protein in SHI-1 cells. SIM also can enhance the chemotherapeutic sensitivity of SHI-1 cells.

Absorption and metabolism characteristics of pristimerin as determined by a sensitive and reliable LC-MS/MS method.

In this research, a sensitive and reliable LC-MS/MS method was developed and applied to determine the concentration of pristimerin in rat plasma, cell incubation media and metabolism incubation mixtures. The absolute oral bioavailability of pristimerin is 28.4% at a dose of 1 mg·kg(-1), and the bioavailability was poor. The bidirectional transport of pristimerin across Caco-2 cells was studied in vitro. A markedly higher transport of pristimerin across Caco-2 cells was observed in the basolateral-to-apical direction and was abrogated in the presence of the P-gp inhibitor, verapamil. The result indicated that P-gp might be involved in the transport of pristimerin in intestine. The phase I and phase II metabolic stability was also investigated using human liver microsomes (HLM) and S9 fractions, respectively. Pristimerin was stable in S9 fractions but metabolized in HLM with a half-life of 20.4 min, which indicated that pristimerin could be mainly metabolized by phase I enzymes. In conclusion, the absolute oral bioavailability of pristimerin in plasma, transport across Caco-2 cell monolayers, and metabolic stability in HLM and S9 fractions were systematically investigated by using a sensitive and reliable LC-MS/MS method.

[Comparative investigation of molecules releasing from intra-hollow calcium-alginate capsules using fluorimetry].

In this paper, the authors employ three different types of dye molecules, Nile red, Rhodamine 6G, fluorescein and a fluorescent protein-R-phycoerythrin (R-PE). The Rhodamine 6G is positively charged molecules, fluorescein is negatively charged molecules, and Nile red is neutral molecules. The R-phycoerythrins have either a net positive or negative charge which is balanced at the isoelectric point (4.22). It is negatively charged molecules also under our experimental condition. The Nile red, rhodamine 6G, fluorescein and R-phycoerythrin are trapped into alginate calcium hollow capsule respectively. The diffusion processes of those molecules from calcium alginate capsule to solution are measured based on a fluorescence method. The results indicate that electrical characteristics of encapsulated molecules have effect on their diffusion behaviors. The positively charged rhodamine 6G is well accordance with a model of control release from porous polymer membranes. The neutral molecules not only can be released from porous polymer framework, they also can directly dissolve out through polymer membrane. The electrostatic repulsion between fluorescein and negatively charged calcium alginate membranes will accelerate the molecular motion, which is propitious to molecules directly dissolving out through polymer membrane. Based on Fick's law of diffusion, R-PEs can be releases from porous polymer framework It shows the longest equilibrium time. Comparing neutral molecules, negatively and positively charge molecules show the stronger interaction on electric polymer membrane, which results in that the diffusion coefficients of rhodamine 6G and fluorescein are less than that of neutral molecule Nile red. The consequences obtained here should readily explain analogous control releasing behaviors of other functional molecules.