Psoralen attenuates cigarette smoke extract-induced inflammation by modulating CD8+ T lymphocyte recruitment and chemokines via the JAK2/STAT1 signaling pathway.

Chronic obstructive pulmonary disease (COPD) is a respiratory inflammatory disease. Psoralen (PSO) is the main pharmacological component identified from Bu-Shen-Fang-Chuan formula which has been traditionally used in treatment of COPD, yet its efficacy in COPD inflammation were unreported. In this study, we aimed to elucidate the anti-inflammatory potential of PSO in COPD and unravel the underlying mechanisms, focusing on T lymphocyte recruitment and the modulation of chemokines, namely monokine induced by interferon-gamma (CXCL9), interferon inducible protein 10 (CXCL10), and interferon inducible T-Cell alpha chemoattractant (CXCL11). In vitro, RAW264.7 was stimulated by interferon (IFN)-γ + cigarette smoke extract (CSE) and were treated with PSO (2.5, 5, 10 µM), then the levels of chemokines and the activation of Janus kinase (JAK)/Signal transducer and activator of transcription 1 (STAT1) pathway were analyzed by real time PCR and western blot. In vivo, a murine model was established by intraperitoneal injection of CSE on day 1, 8, 15, and 22, then treated with PSO (10 mg/kg). Our experiments in vitro illustrated that PSO reduced the levels of CXCL9, CXCL10, and CXCL11, and decreased the protein phosphorylation levels of JAK2 and STAT1. Additionally, PSO effectively improved inflammatory infiltration and decreased the proportion of CD8+ T cells in CSE-exposed mice. Furthermore, PSO reduced the levels of CXCL9, CXCL10, and CXCL11 in bronchoalveolar lavage fluid (BALF) and lung tissue, and decreased the protein phosphorylation levels of JAK2 and STAT1. In conclusion, our results revealed the therapeutic potential of PSO for COPD inflammation, possibly mediated through the regulation of CD8+ T cell recruitment and chemokines via the JAK2/STAT1 signaling pathway.

Target analysis and identification of curcumin against vascular calcification.

To investigate the mechanism of curcumin (CUR) on vascular calcification (VC), we screen for common targets of CUR and atherosclerosis and verify the targets genes in vivo and in vitro experiments. The common targets of CUR and AS were screened and obtained using different databases. These target genes were analyzed by GO and KEGG pathway enrichment analysis. PPI network analysis was performed and to analyze the key targets. A rat VC model was constructed and CUR was fed for three weeks. The changes of vascular structure and calcium salt deposition were observed in H&E and Von Kossa staining. Further, the expression of these target proteins was detected in the primary VSMCs of VC. The 31 common targets were obtained. GO functional enrichment analysis obtained 1284 terms and KEGG pathway enriched 66 pathways. The key genes were identified in the cytoHubba plugin. The molecular docking analysis showed that CUR bound strongly to EGFR, STAT3 and BCL2. The animal experiments showed the deposition calcium salt reduced by the CUR administration. These proteins BMP2, RUNX2, EGFR, STAT3 and BAX expression were upregulated in VC group and CUR attenuated the upregulated expression. The signal protein Akt and p65 expression increased in VC group and decreased in CUR group. We identified some common target genes of CUR and AS and identified these key genes. The anti-VC effect of CUR was associated with the inhibition of upregulation of EGFR, STAT3 and RUNX2 expression in VSMCs.

Icariside â ¡ attenuates bleomycin-induced pulmonary fibrosis by modulating macrophage polarization.

ETHNOPHARMACOLOGICAL RELEVANCE: Numerous studies have provided evidence supporting the significant roles of icariin, in the prevention of multiple chronic diseases like diabetes, liver fibrosis, cardiac fibrosis, renal fibrosis, and pulmonary fibrosis. In particular, Icariside II (ISE II), a prominent flavonoid glycoside derived from Epimedium brevicornum Maxim, the principal metabolite of icariin, has demonstrated noteworthy anti-inflammatory and anti-oxidant properties, along with its ability to protect against lung remodeling. However, the research exploring ISE Ⅱ's application in treating pulmonary fibrosis remains limited. AIM OF THE STUDY: The aim of this study was to assess the therapeutic efficacy of ISE II in models of pulmonary fibrosis, while also investigating its potential mechanisms of action in cell signaling pathways. MATERIALS AND METHODS: An in vitro model of pulmonary fibrosis was established by treating NIH-3T3 cells with transforming growth factor-ß1 (TGF-ß1). Western blot, RT-qPCR, and scratch test were performed to assess the effect of ISE Ⅱ. In addition, a murine model of pulmonary fibrosis was induced by intratracheal instillation of bleomycin, and the therapeutic effect of ISE Ⅱ was tested by orally administering ISE Ⅱ at a dose of 10 mg/kg. Three weeks later, lung function, micro-CT, hydroxyproline content, pathological staining, and cytokines detection of BALF or serum were used to assess the anti-fibrosis effects of ISE Ⅱ. Next, immunofluorescence staining, flow cytometry, and in vivo transcriptomics were used to investigate the underlying mechanisms of action. RESULTS: Our data revealed a significant inhibitory effect of ISE Ⅱ on the upregulation of α-smooth muscle actin (α-SMA) and collagen production induced by TGF-ß1 in fibroblasts. Meanwhile, ISE Ⅱ exerted a therapeutic effect against bleomycin-induced pulmonary fibrosis in mice by improving lung function, decreasing collagen deposition, and reducing the expression of interleukin (IL)-1ß, tumor necrosis factor α (TNF-α), TGF-ß1 and platelet-derived growth factor (PDGF) in serum and bronchoalveolar lavage fluid (BALF). Additionally, ISE Ⅱ treatment effectively attenuated the infiltration of M2 macrophages, concurrently downregulating the expression level of M2 marker genes, such as CD206, arginase-1(Arg-1), and Chitinase-Like Protein 3 (YM-1). Importantly, we observed a statistically significant reduction in the M2 phenotype of interstitial macrophages (IMs). However, the impact of ISE Ⅱ on the M2 polarization of alveolar macrophages (AMs) did not reach statistical significance. Lastly, transcriptome sequencing results suggested that the anti-pulmonary fibrosis effects of ISE Ⅱ may be mediated by the suppression of the WNT/ß-catenin signaling pathway, which modulated M2 polarization in macrophages and contributed to the amelioration of pulmonary fibrosis. By immunohistochemical analysis, it was verified that ISE Ⅱ treatment dramatically inhibited the activation of ß-catenin in fibrosis murine. CONCLUSION: Our findings indicated that ISE Ⅱ exerted anti-fibrotic effects by inhibiting pro-fibrotic macrophage polarization. The underlying mechanism of action might be mediated by modulating the WNT/ß-catenin signaling pathway to inhibit the M2 program in IMs.

Identification of key genes and pathways in atherosclerosis using integrated bioinformatics analysis.

BACKGROUND: Atherosclerosis (AS) is a chronic inflammatory disease that might induce severe cardiovascular events, such as myocardial infarction and cerebral infarction. These risk factors in the pathogenesis of AS remain uncertain and further research is needed. This study aims to explore the potential molecular mechanisms of AS by bioinformatics analyses. METHODS: GSE100927 gene expression profiles, including 69 AS samples and 35 healthy controls, were downloaded from Gene Expression Omnibus database and indenfied for key genes and pathways in AS. RESULTS: A total of 443 differentially expressed genes (DEGs) between control and AS were identified, including 323 down-regulated genes and 120 up-regulated genes. The Gene ontology terms enriched by the up-regulated DEGs were associated with the regulation of leukocyte activation, endocytic vesicle, and cytokine binding, while the down-regulated DEGs were associated with negative regulation of cell growth, extracellular matrix, and G protein-coupled receptor binding. KEGG pathway analysis showed that the up-regulated DEGs were enriched in Osteoclast differentiation and Phagosome, while the down-regulated DEGs were enriched in vascular smooth muscle contraction and cGMP-PKG signaling pathway. Using the modular analysis of Cytoscape, we identified 3 modules mainly involved in Leishmaniasis and Osteoclast differentiation. The GSEA analysis showed the up-regulated gene sets were enriched in the ribosome, ascorbated metabolism, and propanoate metabolism. The LASSO Cox regression analysis showed the top 3 genes were TNF, CX3CR1, and COL1R1. Finally, we found these immune cells were conferred significantly higher infiltrating density in the AS group. CONCLUSIONS: Our data showed the pathway of Osteoclast differentiation and Leishmaniasis was involved in the AS process and we developed a three-gene model base on the prognosis of AS. These findings clarified the gene regulatory network of AS and may provide a novel target for AS therapy.

Increased Neuromedin B is Associated with a Favorable Prognosis in Glioblastoma.

BACKGROUND: Neuromedin B (NMB) is a neuropeptide that plays a key role in many physiological processes and is involved in the pathology of various diseases. Increased levels of NMB have been reported in solid tumors. Therefore, we investigated the prognostic value of NMB in glioblastoma (GBM). METHODS: Expression profiles of NMB mRNA were investigated in GBM and normal tissues using data from the cancer genome atlas (TCGA). NMB protein expression was obtained using data from the Human Protein Atlas. Receiver operating characteristic (ROC) curves were evaluated in GBM and normal tissues. The survival effect of NMB in GBM patients was evaluated using the Kaplan-Meier method. Protein-protein interaction networks were constructed using STRING, and the functional enrichment analyses were performed. The relationship between NMB expression and tumor-infiltrating lymphocytes was analyzed using the Tumor Immune Estimation Resource (TIMER) and the Tumor-Immune System Interaction database (TISIDB). RESULTS: NMB was overexpressed in GBM relative to normal biopsy specimens. The ROC analysis showed that the sensitivity and specificity of NMB in GBM were 96.4% and 96.2%, respectively. Kaplan-Meier survival analysis showed that GBM patients with high NMB expression had a better prognosis than those with low NMB expression (16.3 vs. 12.7 months, p = 0.002). Correlation analysis showed that NMB expression was associated with tumor-infiltrating lymphocytes and tumor purity. CONCLUSIONS: High expression of NMB was associated with increased GBM patient survival. Our study indicated that the NMB expression may be a biomarker for prognosis and that NMB may be an immunotherapy target in GBM.

Favorable Propylene-Incorporated Terpolymerization of Ethylene with CO Mediated by Cationic [P,O]-Pd and Ni Complexes.

Commercial polyketone materials are generally produced by palladium-catalyzed terpolymerization of ethylene and α-olefin with carbon monoxide (CO), and rare examples were reported regarding the incorporation of propylene into an ethylene/CO copolymer chain using a cost-effective nickel catalyst. In this study, we have developed a series of [P,O]-type cationic Pd and Ni complexes supported by a diphosphazane monoxide (PNPO) platform, and the electronic and steric effect on phosphine, amine, and phosphine oxide moieties is systematically investigated for terpolymerization in terms of activity, propylene/CO (C3) incorporation, and molecular weight control. It is observed that the melting temperature (Tm) is proportional to the number of C3 incorporations present in the polymer chain, and the incorporated propylene does not affect the degradation temperature substantially, thus broadening the processing temperature window of the resultant polyketones. Notably, in comparison with dppp-type catalysts, PNPO catalysts exhibited a higher preference for propylene consumption, which is of great importance for making more efficient use of α-olefin resources.

Migration study of phenolic endocrine disruptors from pacifiers to saliva simulant by solid phase microextraction with amino-functionalized microporous organic network coated fiber.

Parabens, bisphenols, and triclosan are used in many baby products, including pacifiers. However, the migration through oral saliva will result in a potential health risk. The present study proposes a sensitive and simple method for the analysis of these chemicals in saliva simulants by solid phase microextraction (SPME) with amino-functionalized microporous organic network (MON-NH2) coated fiber. The MON-NH2 showed an excellent adsorption ability for phenolic compounds. The adsorption isotherm fitted the Langmuir isotherm model and the adsorption kinetics followed the pseudo second-order model. The developed SPME method exhibited wide linear ranges (0.005-500 µg/L), good linearity, low limits of quantitation (0.005 µg/L), great recoveries (87.0-112.5 %), and excellent precision (RSD < 8.3 % for intra-day and RSD < 13.7 % for inter-day). Mathematical models based on Fick's second law were applied to predict migration from pacifiers into saliva simulants and a good fit between theoretical and experimental migration results was found. The daily exposure assessment results indicated that these chemicals in pacifiers do not pose unacceptable health risks to infants. However, exposure risks still should be monitored and appropriate precautions are still needed to protect infants from exposure to these chemicals.

Cationic P,O-Coordinated Nickel(II) Catalysts for Carbonylative Polymerization of Ethylene: Unexpected Productivity via Subtle Electronic Variation.

Transition-metal-catalyzed copolymerization of ethylene with carbon monoxide affords polyketones materials with excellent mechanical strength, photodegradability, surface and barrier properties. Unlike the widely used and rather expensive Pd catalysts, Ni-catalyzed carbonylative polymerization is very difficult since the strong binding affinity of CO to Ni deactivates the highly electrophilic metal center easily. In this study, various cationic P,O-coordinated Ni complexes were synthesized using the electronic modulation strategy, and the catalyst with strong electron-donating substituents exhibits an excellent productivity of 104  g polymer (g Ni)-1 , which represents a rare discovery that a Ni complex could operate with such exceptional efficiency in comparison with Pd catalysts. Notably, those Ni catalysts were also efficient for terpolymerization of ethylene, propylene with CO for producing commercial polyketone materials with low melting temperatures and easy processibility.

Hollow fiber membrane-protected amino/hydroxyl bifunctional microporous organic network fiber for solid-phase microextraction of bisphenols A, F, S, and triclosan in breast milk and infant formula.

Bisphenols and triclosan have been used in various products, and exposure to these chemicals may affect human health. The present study proposes a sensitive method for the determination of bisphenols A, F, S, and triclosan. The fiber was coated by amino/hydroxyl bifunctional microporous organic network and protected by polyvinylidene fluoride hollow fiber membrane for direct immersion solid phase microextraction. The limit of detection was 0.005 µg/L (µg/kg), and the recoveries were in the range of 76.7% to 107.5% (87.4% to 107.6%) for breast milk (infant formula), with intra-day and inter-day precisions <10.5% (7.3%) and 13.6% (8.4%), respectively. Fiber-to-fiber reproducibility of < 9.5% and a lifespan of >100 cycles were obtained. The 95th percentile estimated daily intake of total bisphenols was close to temporary tolerable daily intake for infants fed by human milk, which highlighted the needs for further attention on human exposure to BPA and its substitutes.

Determination of pesticide residues in chilli and Sichuan pepper by high performance liquid chromatography quadrupole time-of-flight mass spectrometry.

In this study, a multi-component analytical method for the detection of pesticide residues in chilli and Sichuan pepper by high performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (LC-Q-TOF/MS) was developed and validated. The sample preparation is based on an extraction step with acetonitrile followed by a cleanup step using primary secondary amine, C18, graphitized carbon black and anhydrous magnesium sulfate. Values of matrix effects ranged from -55.8 to 26.0 % for chilli, and -69.1 to 24.0 % for Sichuan pepper. The LOQ of ≤ 5 µg kg-1 was achieved for all the target pesticides. Applying the method to real samples, some pesticides were found at high concentrations, which were beyond the MRL set by the EU. The results showed that the developed method could be used for the quantitative analysis of target pesticides and non-target screening for potential metabolites in chilli and Sichuan pepper.

Intra-muscular extraskeletal myxoid chondrosarcoma of the thigh: a case report.

Soft tissue sarcomas are mesenchymal tumors that account for about 1% of all malignancies. We retrospectively analyzed a rare case of a painful intra-muscular extraskeletal myxoid chondrosarcoma in the thigh of a 35-year-old man, that had undergone excision. Histological and immunohistochemical analysis of the mass revealed extraskeletal myxoid chondrosarcoma. The patient proceeded to radiotherapy and chemotherapy after curative surgery and had a good outcome.

Involvement of phospholipase PLA2 in production of cellulase and xylanase by Penicillium oxalicum.

Phospholipases play vital roles in immune and inflammatory responses in mammals and plants; however, knowledge of phospholipase functions in fungi is limited. In this study, we investigated the effects of deleting predicted phospholipase genes on cellulase and xylanase production, and morphological phenotype, in Penicillium oxalicum. Individual deletion of nine of the ten predicted phospholipase genes resulted in alteration of cellulase and xylanase production, and the morphological phenotypes, to various degrees. The mutant ∆POX07277 lost 22.5 to 82.8% of cellulase (i.e., filter paper cellulase, carboxymethylcellulase, and p-nitrophenyl-ß-cellobiosidase) and xylanase production, whereas p-nitrophenyl-ß-glucopyranosidase production increased by 5.8-127.8 fold. POX07277 (P. oxalicum gene No. 07277) was predicted to encode phospholipase A2 and was found to negatively affect the sporulation of P. oxalicum. Comparative transcriptomic and quantitative reverse transcription-PCR analysis indicated that POX07277 dynamically affected the expression of cellulase and xylanase genes and the regulatory genes for fungal sporulation, under micro-crystalline cellulose induction. POX07277 was required for the expression of the known regulatory gene PoxCxrB (cellulolytic and xylanolytic regulator B in P. oxalicum), which is involved in cellulase and xylanase gene expression in P. oxalicum. Conversely, POX07277 expression was regulated by PoxCxrB. These findings will aid the understanding of phospholipase functions and provide novel insights into the mechanism of fungal cellulase and xylanase gene expression. KEY POINTS : • The roles of phospholipases were investigated in Penicillium oxalicum. • POX07277 (PLA2) is required for the expression of cellulase and xylanase genes. • PoxCxrB dynamically regulated POX07277 expression.

ARTP/EMS-combined multiple mutagenesis efficiently improved production of raw starch-degrading enzymes in Penicillium oxalicum and characterization of the enzyme-hyperproducing mutant.

BACKGROUND: Application of raw starch-degrading enzymes (RSDEs) in starch processing for biofuel production can effectively reduce energy consumption and processing costs. RSDEs are generally produced by filamentous fungi, such as Penicillium oxalicum, but with very low yields, which seriously hampers industrialization of raw starch processing. Breeding assisted by random mutagenesis is an efficient way to improve fungal enzyme production. RESULTS: A total of 3532 P. oxalicum colonies were generated after multiple rounds of mutagenesis, by atmospheric and room-temperature plasma (ARTP) and/or ethyl methanesulfonate (EMS). Of these, one mutant A2-13 had the highest RSDE activity of 162.7 U/mL, using raw cassava flour as substrate, a yield increase of 61.1%, compared with that of the starting strain, OXPoxGA15A. RSDE activity of A2-13 further increased to 191.0 U/mL, through optimization of culture conditions. Increased expression of major amylase genes, including the raw starch-degrading glucoamylase gene, PoxGA15A, and its regulatory gene, PoxAmyR, as well as several single-nucleotide polymorphisms in the A2-13 genome, were detected by real-time reverse transcription quantitative PCR and genomic re-sequencing, respectively. In addition, crude RSDEs produced by A2-13, combined with commercial α-amylase, could efficiently digest raw corn flour and cassava flour at 40 °C. CONCLUSIONS: Overall, ARTP/EMS-combined mutagenesis effectively improved fungal RSDE yield. An RSDE-hyperproducing mutant, A2-13, was obtained, and its RSDEs could efficiently hydrolyze raw starch, in combination with commercial α-amylase at low temperature, which provides a useful RSDE resource for future starch processing.