[Mechanism of bulleyaconitine A in inhibiting bone destruction of rheumatoid arthritis via Src/PI3K/Akt signaling pathway].

Based on the sarcoma receptor coactivator(Src)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) signaling pathway, the mechanism of action of bulleyaconitine A in the treatment of bone destruction of experimental rheumatoid arthritis(RA) was explored. Firstly, key targets of RA bone destruction were collected through GeneCards, PharmGKB, and OMIM databa-ses. Potential targets of bulleyaconitine A were collected using SwissTargetPrediction and PharmMapper databases. Next, intersection targets were obtained by the Venny 2.1.0 platform. Protein-protein interaction(PPI) network and topology analysis were managed by utilizing the STRING database and Cytoscape 3.8.0. Then, Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were conducted in the DAVID database. AutoDock Vina was applied to predict the molecular docking and binding ability of bulleyaconitine A with key targets. Finally, a receptor activator of nuclear factor-κB(RANKL)-induced osteoclast differentiation model was established in vitro. Quantitative real-time polymerase chain reaction(qRT-PCR) was used to detect the mRNA expression levels of related targets, and immunofluorescence and Western blot were adopted to detect the protein expression level of key targets. It displayed that there was a total of 29 drug-disease targets, and Src was the core target of bulleyaconitine A in anti-RA bone destruction. Furthermore, KEGG enrichment analysis revealed that bulleyaconitine A may exert an anti-RA bone destruction effect by regulating the Src/PI3K/Akt signaling pathway. The molecular docking results showed that bulleyaconitine A had better bin-ding ability with Src, phosphatidylinositol-4,5-diphosphate 3-kinase(PIK3CA), and Akt1. The result of the experiment indicated that bulleyaconitine A not only dose-dependently inhibited the mRNA expression levels of osteoclast differentiation-related genes cathepsin K(CTSK) and matrix metalloproteinase-9(MMP-9)(P<0.01), but also significantly reduced the expression of p-c-Src, PI3K, as well as p-Akt in vitro osteoclasts(P<0.01). In summary, bulleyaconitine A may inhibit RA bone destruction by regulating the Src/PI3K/Akt signaling pathway. This study provides experimental support for the treatment of RA bone destruction with bulleyaconitine A and lays a foundation for the clinical application of bulleyaconitine A.

[Mechanism of Yuxuebi Tablets in treating synovial inflammation in rheumatoid arthritis based on transcriptomics].

This study investigated the mechanism of Yuxuebi Tablets(YXB) in the treatment of synovial inflammation in rheumatoid arthritis(RA) based on transcriptomic analysis. Transcriptome sequencing technology was employed to analyze the gene expression profiles of joint tissues from normal rats, collagen-induced arthritis(CIA) rats(an RA model), and YXB-treated rats. Common diffe-rentially expressed genes(DEGs) were subjected to Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses. RA synovial inflammation-related target genes were retrieved from the OMIM and GeneCards databases. Venny 2.1 software was used to identify the intersection of YXB target genes and RA synovial inflammation-related target genes, and GO and KEGG enrichment analyses were performed on the intersecting target genes. Immunohistochemistry was used to assess the protein expression levels of the inflammatory factors interleukin-1ß(IL-1ß) and tumor necrosis factor-α(TNF-α) in rat joint tissues. Western blot analysis was employed to measure the expression levels of key proteins in the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) signaling pathway. A total of 2 058 DEGs were identified by intersecting the genes from the normal group vs model group and the model group vs YXB treatment group. A search in OMIM and GeneCards databases yielded 1 102 RA synovial inflammation-related target genes. After intersecting with the DEGs in the YXB treatment group, 204 intersecting target genes were identified, primarily involving biological processes such as immune response, signal transduction, and inflammatory response; cellular components including plasma membrane, extracellular space, and extracellular region; molecular functions like protein binding, identical protein binding, and receptor binding. These target genes were mainly enriched in signaling pathways such as PI3K/Akt, cytokine-cytokine receptor interaction, and Janus kinase/signal transducer and activator of transcription(JAK/STAT). Western blot results showed that YXB at low, medium, and high doses could significantly inhibit the expression levels of key proteins in the PI3K/Akt signaling pathway in rat joint tissues in a dose-dependent manner. Immunohistochemistry further confirmed these findings, showing that YXB not only suppressed the protein expression levels of the inflammatory factors IL-1ß and TNF-α in the joint synovial tissues of CIA rats, but also inhibited p-Akt protein expression. In conclusion, this study used transcriptomic analysis to uncover the key mechanisms of YXB in inhibiting synovial inflammation and alleviating the progression of RA, with a focus on its role in suppressing the PI3K/Akt signaling pathway.

Vertical migration behavior simulation and prediction of Pb and Cd in co-contaminated soil around Pb-Zn smelting slag site.

Heavy metal migration in soil poses a serious threat to the soil and groundwater. Understanding the migration pattern of heavy metals (HMs) under different factors could provide a more reasonable position for pollution evaluation and targetoriented treatment of soil heavy metal. In this study, the migration behavior of Pb and Cd in co-contaminated soil under different pH and ionic strength (NaCl concentration) was simulated using convective dispersion equation (CDE). We predicted the migration trends of Pb and Cd in soils after 5, 10, and 20 years via PHREEQC. The results showed that the migration time of Cd in the soil column experiment was about 60 days faster than that of Pb, and the migration trend was much steeper. The CDE was proved to describe the migration behavior of Pb and Cd (R2 > 0.75) in soil. The predicted results showed that Cd migrated to 15-20 cm of soil within 7 years and Pb stayed mainly in the top 0-6 cm of soil within 5 years as the duration of irrigation increased. Overall, our study is expected to provide new insight into the migration of heavy metal in soil ecosystems and guidance for reducing risk of heavy metal in the environment.

[Intervention effect of Qufeng Gutong Cataplasm on myofascial pain syndrome in rats and its mechanism].

This paper aims to investigate the intervention effect of Qufeng Gutong Cataplasm(QFGT) on myofascial pain syndrome(MPS) in rats and to preliminarily explain its mechanism from the perspective of improving muscle inflammation and pain. Male SD rats were divided into 6 groups, namely normal group, model group, positive control drug(Huoxue Zhitong Ointment, HXZT) group, and low, medium, and high-dose QFGT groups(75, 150, and 300 mg·d~(-1)). The rat model of MPS was established by striking combined with centrifugation for 8 weeks, during which QFGT and HXZT were used for corresponding intervention. Standard VonFrey fiber was used to evaluate the mechanical pain threshold, and acetone was used to detect the cold pain threshold. The electrophysiological activity of muscle at trigger point was detected, and the electromuscular analysis of trigger point was performed. CatWalk gait analyzer was used to detect pain-induced gait adaptation changes. The hematoxylin-eosin(HE) staining was used to observe the pathological changes in muscle and skin tissues at the trigger point of rats. Immunohistochemistry was used to detect the expression of capsaicin receptor transient receptor potential vanilloid 1(TRPV1) in muscle tissues and interleukin(IL)-33 in skin tissues at the trigger point. The protein expression levels of TRPV1, protein kinase B(Akt), phosphorylated protein kinase B(p-Akt), IL-1ß, and tumor necrosis factor-α(TNF-α) in muscle tissues at the trigger point were detected by Western blot. The results showed that as compared with the model group, the mechanical pain threshold and cold pain threshold of rats in other groups were increased after treatment with QFGT. The spontaneous electromyography(EMG) activity was observed in the model group, but QFGT alleviated the EMG activity in a dose-dependent manner. Gait analysis showed that standing duration, average intensity, swing speed, maximum contact point, maximum contact area, paw print length, paw print width, and paw print area were significantly improved in all QFGT groups. Pathological results showed that the disorder of muscle arrangement at the trigger point was decreased, muscle fiber adhesion and atrophy were reduced, and inflammatory cell infiltration was alleviated after treatment with QFGT. In addition, QFGT and HXZT both inhibited the protein expression of TRPV1, PI3K, Akt, p-Akt, IL-1ß, and TNF-α in the muscle tissues of rats with MPS. However, there was no significant difference in the pathological structure and expression of IL-33 in the treated skin as compared with the normal group. The related results have proved that QFGT can inhibit the release of inflammatory factors by inhibiting the TRPV1/PI3K/Akt signaling pathway in the muscle trigger point of rats with MPS and finally attenuate the atrophy and adhesion of local muscles and inflammatory infiltration, thereby relieving the muscle pain of rats with MPS, and local administration has no skin irritation.

[Mechanism of artesunate on bone destruction in experimental rheumatoid arthritis based on transcriptomics and network pharmacology].

The present study investigated the mechanism of artesunate in the treatment of bone destruction in experimental rheumatoid arthritis(RA) based on transcriptomics and network pharmacology. The transcriptome sequencing data of artesunate in the inhibition of osteoclast differentiation were analyzed to obtain differentially expressed genes(DEGs). GraphPad Prism 8 software was used to plot volcano maps and heat maps were plotted through the website of bioinformatics. GeneCards and OMIM were used to collect information on key targets of bone destruction in RA. The DEGs of artesunate in inhibiting osteoclast differentiation and key target genes of bone destruction in RA were intersected by the Venny 2.1.0 platform, and the intersection target genes were analyzed by Gene Ontology(GO)/Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment. Finally, the receptor activator of nuclear factor-κB(RANKL)-induced osteoclast differentiation model and collagen-induced arthritis(CIA) model were established. Quantitative real time polymerase chain reaction(q-PCR), immunofluorescence, and immunohistochemistry were used to verify the pharmacological effect and molecular mechanism of artesunate in the treatment of bone destruction in RA. In this study, the RANKL-induced osteoclast differentiation model in vitro was established and intervened with artesunate, and transcriptome sequencing data were analyzed to obtain 744 DEGs of artesunate in inhibiting osteoclast differentiation. A total of 1 291 major target genes of bone destruction in RA were obtained from GeneCards and OMIM. The target genes of artesunate in inhibiting osteoclast differentiation and the target genes of bone destruction in RA were intersected to obtain 61 target genes of artesunate against bone destruction in RA. The intersected target genes were analyzed by GO/KEGG enrichment. According to the results previously reported, the cytokine-cytokine receptor interaction signaling pathway was selected for experimental verification. Artesunate intervention in the RANKL-induced osteoclast differentiation model showed that artesunate inhibited CC chemokine receptor 3(CCR3), CC chemokine receptor 1(CCR1) and leukemia inhibitory factor(LIF) mRNA expression in osteoclasts in a dose-dependent manner compared with the RANKL-induced group. Meanwhile, the results of immunofluorescence and immunohistochemistry showed that artesunate could dose-dependently reduce the expression of CCR3 in osteoclasts and joint tissues of the CIA rat model in vitro. This study indicated that artesunate regulated the CCR3 in the cytokine-cytokine receptor interaction signaling pathway in the treatment of bone destruction in RA and provided a new target gene for the treatment of bone destruction in RA.

Contamination characteristics and source analysis of potentially toxic elements in dustfall-soil-crop systems near non-ferrous mining areas of Yunnan, southwestern China.

Potentially toxic elements (PTEs) in the dustfall-soil-crop system pose a serious threat to the ecological environment and agricultural production. However, there is still a knowledge gap in terms of better understanding the distinctive sources of PTEs by integrating various models and technologies. In this study, we comprehensively investigated the concentrations, distribution, and sources of seven PTEs in a dustfall-soil-crop system (424 samples in total) near a typical non-ferrous mining area, using absolute principal component score/multiple linear regression (APCS/MLR) combined with X-ray diffraction (XRD) and microscopy techniques. Our results showed that the mean values of As, Cd, Cr, Cu, Ni, Pb, and Zn in the soils were 211, 14, 105, 91, 65, 232, and 325 mg/kg, respectively. These values were significantly higher than the background soil values in Yunnan. Except for Ni and Cr, all elements in the soil were significantly higher than the screening values of agricultural lands in China. The spatial distribution of PTE concentrations was similar among the three media. The ACPS/MLR, XRD, and microscopy analyses further indicated that soil PTEs mainly originated from industrial activities (37 %), vehicle emissions and agricultural activities (29 %), respectively. Dustfall PTEs mainly originated from vehicle emissions and industrial activities, accounting for 40 % and 37 %, respectively. Crop PTEs mainly originated from vehicle emissions and soil (57 %), and agricultural activities (11 %), respectively. PTEs seriously threaten the safety of agricultural products and the ecological environment once they settle from the atmosphere to soil and crop leaves, further accumulate in crops, and spread through the food chain. Therefore, our study provides scientific evidence for government regulators to control PTE pollution and reduce their environmental risks in dustfall-soil-crop systems.

Characteristics and source analysis of potentially toxic elements pollution in atmospheric fallout around non-ferrous metal smelting slag sites-taking southwest China as an example.

More accurate source analysis of potentially toxic elements (PTEs) in atmospheric fallout that endanger biodiversity and human health remains needed. This study determined the concentrations of seven PTEs, including Pb, Cd, As, Cu, Zn, Ni, and Cr, by inductively coupled plasma mass spectrometry (ICP-MS), and the sources of PTE pollution were quantified using multivariate statistical analysis, including principal component analysis (PCA), cluster analysis (CA), and Pearson correlation analysis, and Moran index was applied for mutual verification and supplementation. PCA and CA revealed that the same mixed sources of Pb, Cd, As, Cu, and Zn were found in the atmospheric dust fall in the study area, while Ni and Cr had the same source of pollution. Pearson correlation analysis provided that there were strong correlations between Pb-Cd, Pb-As, Pb-Cu, Cd-As, Cd-Cu, As-Cu, and Ni-Cr, indicating commonality between the two sources of heavy metal pollution. Additionally, the Moran index showed that strong spatial correlations were observed between Pb, Cd, As, Cu, and Zn, whose sources were mainly related to non-ferrous metal processing smelter smelting slag sites and an environmental company in the study area. However, no spatial correlation was found between Ni and Cr, which mainly originated from the local geological background.

Roles of extracellular polymeric substances in uranium immobilization by anaerobic sludge.

The specific roles of extracellular polymeric substances (EPS) and how factors influenced EPS's roles during U(VI) immobilization are still unclear. In this study, high content of U with the main form of nanoparticles was detected in EPS, accounting for 10-42% of total U(VI) removal. EPS might be utilized as energy source or even as electron donors when external carbon source was unavailable. The influencing degree of each experimental parameter to uranium (U) removal process was elucidated. The influential priority to U(IV)/U(VI) ratios in sludge was as follows: acetate, U(VI), and nitrate. The influential priority to total EPS contents was as follows: U(VI), nitrate and acetate. The complex interaction mechanism between U(VI) and EPS in the U immobilization process was proposed, which might involve three ways including biosorption, bioreduction and bioprecipitation. These results indicate important and various roles of EPS in U(VI) immobilization.

[Stromal cells inhibit Kallikrein 7 expression in the epithelial cells of benign prostate hyperplasia].

OBJECTIVE: To investigate the influence of stromal cells on the Kallikrein 7 (KLK7) expression of epithelial cells in benign prostate hyperplasia (BPH). METHODS: We constructed a stromal-epithelial co-culture model after separating the two types of cells from BPH tissues and identifying them by cell morphology and chemiluminescent microparticle immunoassay (CMIA). The expression of KLK7 mRNA was detected by RT-PCR in the epithelial cells with or without the stromal cells, and that of the KLK7 protein (hK7) determined by Western blot. RESULTS: Stromal and epithelial cells were successfully separated and identified, and a stromal-epithelial co-culture model successfully established. RT-PCR showed that the mRNA expression of the KLK7 gene was higher in the epithelial cells co-cultured with stromal cells than in the epithelial cells alone, and the gray value of KLK7 to GAPDH was 1.41 +/- 0.041 in the former and 1.78 +/- 0.10 in the latter (P < 0.01). The results of Western blot were consistent with those of RT-PCR. CONCLUSION: Stromal cells can suppress the expression of the KLK7 gene in the epithelial cells in BPH. KLK7 may be involved in the change of epithelial cells stimulated by stromal cells.

[Characterization of memory CD4+ IL-21+ T cells in human PBMC].

AIM: To characterize IL-21-producing T cells in human PBMCs. METHODS: PBMCs from healthy individuals were stimulated with or without anti-CD3 (OKT3), OKT3 coupled with anti-CD28, or PMA coupled with ionomycin. The cell subsets of IL-21-producing T cells were determined by FACS. PBMCs, CD4+, CD4+CD45RA⁻, CD4+ CD45RA+ or CBMCs were stimulated with PMA coupled with ionomycin. The phenotype of CD4+ IL-21+ T cells and the correlation of IL-21-producing CD4+; T cells with Th1, Th2, Th17 and Th22 cell populations were analyzed by FACS. RESULTS: PMA and ionomycin induced the highest level of IL-21 production among the stimuli tested. CD4+ T cells but not CD8+ T cells mainly expressed IL-21. CD4+ IL-21+ T cells expressed CD45RO instead of CD45RA. Some of the CD4+ IL-21+ T cells expressed CCR6, CCR7 or CXCR5. CD4+ CD45RA⁻ T cells expressed much more IL-21 than CD4+ CD45RA+ T cells. Furthermore, CD4+ T cells from PBMCs but not CBMCs, expressed IL-21. Approximately 24% of CD4+ IL-21+ cells expressed IFN-γ. Less than 10% of CD4+ IL-21+ cells expressed IL-4, IL-17 or IL-22. CONCLUSION: IL-21 is induced from human PBMCs following various polyclonal stimulations. The majority of IL-21-producing cells in PBMCs are memory CD4+ T cells. In addition, some of the CD4+ IL-21+ T cells are distinct from Th1, Th2 and Th17 cells.