Quantitative proteomic analysis of hepatic tissue in allotetraploid hybridized from red crucian carp and common carp identified crucial proteins and pathways associated with metabolism and growth rate.

Allotetraploid is a new species produced by distant hybridization between red crucian carp (Carassius auratus red var., abbreviated as RCC) and common carp (Cyprinus carpio L., abbreviated as CC). There is a significant difference in growth rate between allotetraploid and its parents. However, the underlying molecular mechanism is largely unknown. In this study, to find direct evidence associated with metabolism and growth rate in protein level, we performed quantitative proteomics analysis on liver tissues between allotetraploid and its parents. A total of 2502 unique proteins were identified and quantified by SWATH-MS in our proteomics profiling. Subsequently, comprehensive bioinformatics analyses including gene ontology enrichment analysis, pathway and network analysis, and protein-protein interaction analysis (PPI) were conducted based on differentially expressed proteins (DEPs) between allotetraploid and its parents. The results revealed several significant DEPs involved in metabolism pathways in liver. More specifically, the integrative analysis highlighted that the DEPs ACSBG1, OAT, and LDHBA play vital roles in metabolism pathways including "pentose phosphate pathway," "TCA cycle," and "glycolysis and gluconeogenesis." These could directly affect the growth rate in fresh water fishes by regulating the metabolism, utilization, and exchange of substance and energy. Since the liver is the central place for metabolism activity in animals, we firstly established the comprehensive and quantitative proteomics knowledge base for liver tissue from freshwater fishes, our study may serve as an irreplaceable reference for further studies regarding fishes' culture and growth.

Loss of Wnt16 Leads to Skeletal Deformities and Downregulation of Bone Developmental Pathway in Zebrafish.

Wingless-type MMTV integration site family, member 16 (wnt16), is a wnt ligand that participates in the regulation of vertebrate skeletal development. Studies have shown that wnt16 can regulate bone metabolism, but its molecular mechanism remains largely undefined. We obtained the wnt16-/- zebrafish model using the CRISPR-Cas9-mediated gene knockout screen with 11 bp deletion in wnt16, which led to the premature termination of amino acid translation and significantly reduced wnt16 expression, thus obtaining the wnt16-/- zebrafish model. The expression of wnt16 in bone-related parts was detected via in situ hybridization. The head, spine, and tail exhibited significant deformities, and the bone mineral density and trabecular bone decreased in wnt16-/- using light microscopy and micro-CT analysis. RNA sequencing was performed to explore the differentially expressed genes (DEGs). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis found that the down-regulated DEGs are mainly concentrated in mTOR, FoxO, and VEGF pathways. Protein-protein interaction (PPI) network analysis was performed with the detected DEGs. Eight down-regulated DEGs including akt1, bnip4, ptena, vegfaa, twsg1b, prkab1a, prkab1b, and pla2g4f.2 were validated by qRT-PCR and the results were consistent with the RNA-seq data. Overall, our work provides key insights into the influence of wnt16 gene on skeletal development.

Targeting the STAT3/IL-36G signaling pathway can be a promising approach to treat rosacea.

BACKGROUND: Rosacea is an inflammatory skin disorder characterized by the release of inflammatory mediators from keratinocytes, which are thought to play a crucial role in its pathogenesis. Despite an incidence of approximately 5.5%, rosacea is associated with a poor quality of life. However, as the pathogenesis of rosacea remains enigmatic, treatment options are limited. OBJECTIVES: To investigate the pathogenesis of rosacea and explore new therapeutic strategies. METHODS: Transcriptome data from rosacea patients combined with immunohistochemical staining were used to investigate the activation of STAT3 in rosacea. The role of STAT3 activation in rosacea was subsequently explored by inhibiting STAT3 activation both in vivo and in vitro. The key molecules downstream of STAT3 activation were identified through data analysis and experiments. Dual-luciferase assay and ChIP-qPCR analysis were used to validate the direct binding of STAT3 to the IL-36G promoter. DARTS, in combination with experimental screening, was employed to identify effective drugs targeting STAT3 for rosacea treatment. RESULTS: STAT3 signaling was hyperactivated in rosacea and served as a promoter of the keratinocyte-driven inflammatory response. Mechanistically, activated STAT3 directly bind to the IL-36G promoter region to amplify downstream inflammatory signals by promoting IL-36G transcription, and treatment with a neutralizing antibody (α-IL36γ) could mitigate rosacea-like inflammation. Notably, a natural plant extract (pogostone), which can interact with STAT3 directly to inhibit its activation and affect the STAT3/IL36G signaling pathway, was screened as a promising topical medication for rosacea treatment. CONCLUSIONS: Our study revealed a pivotal role for STAT3/IL36G signaling in the development of rosacea, suggesting that targeting this pathway might be a potential strategy for rosacea treatment.

Twin defect-rich Pt ultrathin nanowire nanozymes alleviate inflammatory skin diseases by scavenging reactive oxygen species.

Nanozymes with superior antioxidant properties offer new hope for treating oxidative stress-related inflammatory skin diseases. However, lacking sufficient catalytic activity or having complex material designs limit the application of current metallic nanozymes in inflammatory skin diseases. Here, we report a simple and effective twin-defect platinum nanowires (Pt NWs) enzyme with multiple mimetic enzymes and broad-spectrum ROS scavenging capability for the treatment of inflammatory skin diseases in mice (including psoriasis and rosacea). Pt NWs with simultaneous superoxide dismutase, glutathione peroxidase and catalase mimetic enzyme properties exhibit cytoprotective effects against ROS-mediated damage at extremely low doses and significantly improve treatment outcomes in psoriasis- and rosacea-like mice. Meanwhile, these ultrasmall sizes of Pt NWs allow the nanomaterials to effectively penetrate the skin and do not produce significant biotoxicity. Therefore, Pt NWs have potential applications in treating diseases related to oxidative stress or inflammation.

Mendelian randomization and transcriptome analysis identified immune-related biomarkers for osteoarthritis.

Background: The immune microenvironment assumes a significant role in the pathogenesis of osteoarthritis (OA). However, the current biomarkers for the diagnosis and treatment of OA are not satisfactory. Our study aims to identify new OA immune-related biomarkers to direct the prevention and treatment of OA using multi-omics data. Methods: The discovery dataset integrated the GSE89408 and GSE143514 datasets to identify biomarkers that were significantly associated with the OA immune microenvironment through multiple machine learning methods and weighted gene co-expression network analysis (WGCNA). The identified signature genes were confirmed using two independent validation datasets. We also performed a two-sample mendelian randomization (MR) study to generate causal relationships between biomarkers and OA using OA genome-wide association study (GWAS) summary data (cases n = 24,955, controls n = 378,169). Inverse-variance weighting (IVW) method was used as the main method of causal estimates. Sensitivity analyses were performed to assess the robustness and reliability of the IVW results. Results: Three signature genes (FCER1G, HLA-DMB, and HHLA-DPA1) associated with the OA immune microenvironment were identified as having good diagnostic performances, which can be used as biomarkers. MR results showed increased levels of FCER1G (OR = 1.118, 95% CI 1.031-1.212, P = 0.041), HLA-DMB (OR = 1.057, 95% CI 1.045 -1.069, P = 1.11E-21) and HLA-DPA1 (OR = 1.030, 95% CI 1.005-1.056, P = 0.017) were causally and positively associated with the risk of developing OA. Conclusion: The present study identified the 3 potential immune-related biomarkers for OA, providing new perspectives for the prevention and treatment of OA. The MR study provides genetic support for the causal effects of the 3 biomarkers with OA and may provide new insights into the molecular mechanisms leading to the development of OA.

Transcriptomic analysis reveals shared gene signatures and molecular mechanisms between obesity and periodontitis.

Background: Both obesity (OB) and periodontitis (PD) are chronic non-communicable diseases, and numerous epidemiological studies have demonstrated the association between these two diseases. However, the molecular mechanisms that could explain the association between OB and PD are largely unclear. This study aims to investigate the common gene signatures and biological pathways in OB and PD through bioinformatics analysis of publicly available transcriptome datasets. Methods: The RNA expression profile datasets of OB (GSE104815) and PD (GSE106090) were used as training data, and GSE152991 and GSE16134 as validation data. After screening for differentially expressed genes (DEGs) shared by OB and PD, gene enrichment analysis, protein-protein interaction (PPI) network construction, GeneMANIA analysis, immune infiltration analysis and gene set enrichment analysis (GSEA) were performed. In addition, receiver operating characteristic (ROC) curves were used to assess the predictive accuracy of the hub gene. Finally, we constructed the hub gene-associated TF-miRNA-mRNA regulatory network. Results: We identified a total of 147 DEGs shared by OB and PD (38 down-regulated and 109 up-regulated). Functional analysis showed that these genes were mainly enriched in immune-related pathways such as B cell receptor signalling, leukocyte migration and cellular defence responses. 14 hub genes (FGR, MNDA, NCF2, FYB1, EVI2B, LY86, IGSF6, CTSS, CXCR4, LCK, FCN1, CXCL2, P2RY13, MMP7) showed high sensitivity and specificity in the ROC curve analysis. The results of immune infiltration analysis showed that immune cells such as macrophages, activated CD4 T cells and immune B cells were present at high infiltration levels in both OB and PD samples.The results of GeneMANIA analysis and GSEA analysis suggested that five key genes (FGR, LCK, FYB1, LY86 and P2RY13) may be strongly associated with macrophages. Finally, we constructed a TF-miRNA-mRNA regulatory network consisting of 233 transcription factors (TFs), 8 miRNAs and 14 mRNAs based on the validated information obtained from the database. Conclusions: Five key genes (FGR, LCK, FYB1, LY86, P2RY13) may be important biomarkers of OB and PD. These genes may play an important role in the pathogenesis of OB and PD by affecting macrophage activity and participating in immune regulation and inflammatory responses.

Investigation of the chemical structure of anti-amyloidogenic constituents extracted from Thamnolia vermicularis.

ETHNOPHARMACOLOGICAL RELEVANCE: Thamnolia vermicularis (Sw.) Schaer (T. vermicularis) is known to have therapeutic effects on various diseases in Southwest China. Recent research has highlighted that T. vermicularis may suppress Aß level and Tau hyperphosphorylation to improve the pathological characteristics of Alzheimer's disease, indicating that it might have the potential to treat Alzheimer's disease. AIM OF THE STUDY: The objective of this study was to evaluate the inhibitory effect of T. vermicularis on the fibril formation of a typical amyloidogenic protein, hen egg white lysozyme (HEWL), and to identify the effective components that could potentially enable an extract of T. vermicularis to be used in the development of novel therapeutic agents. MATERIALS AND METHODS: A water extract was prepared from T. vermicularis (TVWE) and its inhibitory effect on amyloid fibrillation in vitro was investigated using thioflavin T and 8-anilinonapthalene-1-sulfonic acid spectrofluorometric analyses. The anti-amyloidogenic components of TVWE were separated and qualitatively analyzed using thin layer chromatography (TLC), supercritical carbon dioxide extraction (SFE-CO2), and liquid chromatography-mass spectrometry. Finally, the effect of the bioactive components on the structure of HEWL in the early stages of fibrillogenesis was determined by molecular docking simulation. RESULTS: TVWE strongly inhibited the ability of HEWL to form an amyloid fibril, yielding an IC50 of 0.018 mg/mL for the inhibition of fibrillogenesis. The chemical constituents in the various TVWE fractions resolved by TLC were qualitatively identified by liquid chromatography-quadrupole/time-of-flight mass spectrometry (LC-Q-TOF-MS). The target components were predicted by reviewing the existing literature on T. vermicularis, in which the components of T. vermicularis, along with three small molecules (molecular weight: 182) were preliminarily identified. Molecular docking simulation showed that these small molecules were bound to the core region of HEWL, affecting its stability. Finally, the active anti-amyloidogenic components were extracted from whole T. vermicularis using SFE-CO2 and then identified. CONCLUSION: The potential components of TVWE that could prevent HEWL fibrillogenesis were primarily identified using TLC, LC-Q-TOF-MS, and SFE-CO2. The candidate small-molecule compounds were further predicted by combining the LC-Q-TOF-MS results with molecular docking analysis. The effective components of T. vermicularis were extracted using SFE-CO2. Together, these methods could constitute a practical strategy for the isolation and identification of anti-amyloidogenic components from a traditional Chinese medicine.