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Objective To evaluate the therapeutic effect of evodiamine (Evo) on atopic dermatitis (AD) in rat models by regulating the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/cAMP response element binding protein (CREB) signaling pathway. Methods A rat model of AD was established by administration of multiple doses of 2,4-dinitrochlorobenzene (DNCB).The animals were randomly divided into AD group, Evo-low-dose (Evo-L, 5 mg/kg) group, Evo-high-dose (Evo-H, 10 mg/kg) group, Evo-H+H-89 (5 mg/kg) group and dexamethasone (0.1 mg/kg) group. Normal rats were used as the control group and then the degree of skin damage of rats in each group was scored. Abdominal blood was taken to detect the levels of interleukin-4 (IL-4), cAMP, and tumor necrosis factor-α(TNF-α) in serum; Skin lesion tissue was collected to detect pathological change, counting of mast cells, PKA/CREB related protein expression and expression of IL-4 and TNF-α mRNA in the tissue. Results Compared with control group, the level of cAMP in serum, the expression of p-PKA/PKA, and p-CREB/CREB in skin lesions of AD group were reduced, the severity score of skin lesions, level of IL-4 and TNF-α, epidermal thickness, number of mast cells and mRNA expression of IL-4 and TNF-α in skin lesion tissues were all significantly increased (P＜0.05). Compared with AD group, the level of cAMP in serum, the expression of p-PKA/PKA, and p-CREB/CREB in skin lesions in Evo-L group, Evo-H group, and dexamethasone group were increased, the severity score of skin lesions, level of IL-4 and TNF-α, epidermal thickness, number of mast cells, and mRNA expression of IL-4 and TNF-α in skin lesion tissues all reduced (P＜0.05). Compared with the Evo-H group, the level of cAMP in serum, the expression of p-PKA/PKA, and p-CREB/CREB in skin lesions in Evo-H+H-89 group was reduced and the severity score of skin lesion, level of IL-4 and TNF-α, epidermal thickness, number of mast cells, and mRNA expression of IL-4 and TNF-α in skin lesion tissues significantly increased(P＜0.05). Conclusions Evo inhibits inflammatory response and pathological damage through regulation of cAMP/PKA/CREB signaling pathway in AD rat models.

Objective: To investigate the protective effect and mechanism of docosahexaenoic acid-enriched phosphatidylserine (DHA-PS) against cyclophosphamide (CTX)-induced renal injury in mice. Methods: Thirty-two male institute of cancer research (ICR) mice were randomly divided into four groups: control (CON), model (MOD), 50 mg/kg mb DHA-PS (50 DHA-PS), and 100 mg/kg mb DHA-PS (100 DHA-PS), with eight mice in each group. The renal injury model was established by intraperitoneal injection of 80 mg/kg mb CTX. Five days later, the mice in the MOD and DHA-PS groups were gavaged for 7 days with an equal volume of physiological saline and DHA-PS, respectively and then sacrificed. Kidney indexes, serum biochemical indicators, the levels of renal oxidative stress and inflammatory factors, histopathological changes, and non-targeted metabolomics of renal tissues were evaluated. Results: Compared with the MOD group, DHA-PS significantly decreased kidney indexes, creatinine, blood urea nitrogen, cystatin C, and kidney injury molecule-1 levels (P < 0.05), increased the activities of antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) (P < 0.05), reduced the content of malondialdehyde (MDA) (P < 0.05), downregulated the expression of pro-inflammatory cytokines including interleukin (IL)-6, IL-1β, tumor necrosis factor-α (P < 0.05), and upregulated the level of the anti-inflammatory cytokine IL-10 (P < 0.05). Staining results showed a significant restoration of kidney tissue structure in the DHA-PS group. Additionally, the results of renal tissue metabolomics indicated that DHA-PS mitigated CTX-induced metabolic disorders in mouse kidneys mainly by affecting glycerophospholipid metabolism, purine metabolism, and their metabolites. Conclusion: DHA-PS can alleviate CTX-induced renal injury in mice through reducing oxidative stress and inflammatory response, and regulating the glycerophospholipid and purine metabolism pathways.

Abstract Background Paulownia, an ecologically and economically valuable plant species native to China, is notable for its excellent timber quality and strong adaptability. Among them, Paulownia catalpifolia displays the ability to survive in cold climate, a trait associated with northern China. Yet, the molecular information for its cold-tolerance has not been explored. This study was to investigate the changes in physiological indices and transcript levels of P. catalpifolia following cold exposure, which could provide evidence for revealing whether there were differences in the genetic basis of inducing physiological perturbations between moderate low temperature (MLT) and extreme low temperature (ELT). Results The detection of physiological indices under diverse degrees of chilling stress showed similar patterns of alteration. Enhanced accumulation of osmoregulatory substances, such as soluble sugar and soluble protein, were more conducive under ELT compared to MLT in P. catalpifolia. Moreover, we observed leaf wilting symptoms distinctly after exposure to ELT for 48 h, while this effect was not obvious after MLT exposure for 48 h. Comparative transcriptomic analysis between MLT and ELT demonstrated 13,688 differentially expressed genes (DEGs), most of them appeared after 12 h and 48 h of treatment. GO and KEGG analyses elucidated prominent enrichment in aromatic-L-amino-acid decarboxylase activity term and carbohydrate metabolism pathways. Therefore, it was speculated that the DEGs involved in the above processes might be related to the difference in the contents of soluble protein and soluble sugar between MLT and ELT. Time series clustering analyses further highlighted several key genes engaged in the ‘Glycosyltransferases’, ‘Galactose metabolism’ and ‘Starch and sucrose metabolism’ pathways as well as the ‘tyrosine decarboxylase activity’ term. For instance, cellulose synthase-like A (CLSA2/9), raffinose synthase (RafS2), β-amylase (BAM1) and tyrosine/DOPA decarboxylase (TYDC1/2/5) genes, diverging in their expression trends between MLT and ELT, might significantly affect the soluble sugar and soluble protein abundance within P. catalpifolia. Conclusion Between MLT and ELT treatments, partial overlaps in response pathways of P. catalpifolia were identified, while several genes regulating the accumulation of osmotic adjustment substances had disparate expression patterns. These findings could provide a novel physiological and molecular perspective for P. catalpifolia to adapt to complex low temperature habitats.

Abstract Objectives Rheumatoid arthritis (RA) is a chronic autoimmune disease with complex causes and recurrent attacks that can easily develop into chronic arthritis and eventually lead to joint deformity. Our study aims to elucidate potential mechanism among control, new-onset RA (NORA) and chronic RA (CRA) with multi-omics analysis. Methods A total of 113 RA patients and 75 controls were included in our study. Plasma and stool samples were obtained for 16S rRNA sequencing, internally transcribed spacer (ITS) sequencing and metabolomics analysis. And PBMCs were obtained for RNA sequencing. We used three models, logistic regression, least absolute shrinkage and selection operator (LASSO), and random forest, respectively, to distinguish NORA from CRA, and finally we validated model performance using an external cohort of 26 subjects. Results Our results demonstrated intestinal flora disturbance in RA development, with significantly increased abundance of Escherichia-Shigella and Proteobacteria in NORA. We also found that the diversity was significantly reduced in CRA compared to NORA through fungi analysis. Moreover, we identified 29 differential metabolites between NORA and CRA. Pathway enrichment analysis revealed significant dysregulation of glycerophospholipid metabolism and phenylalanine metabolism pathways in RA patients. Next, we identified 40 differentially expressed genes between NORA and CRA, which acetylcholinesterase (ACHE) was the core gene and significantly enriched in glycerophospholipid metabolism pathway. Correlation analysis showed a strong negatively correlation between glycerophosphocholine and inflammatory characteristics. Additionally, we applied three approaches to develop disease classifier models that were based on plasma metabolites and gut microbiota, which effectively distinguished between new-onset and chronic RA patients in both discovery cohort and external validation cohort. Conclusions These findings revealed that glycerophospholipid metabolism plays a crucial role in the development and progression of RA, providing new ideas for early clinical diagnosis and optimizing treatment strategies.

By choosing appropriate candidate reference genes (CRGs) and standardizing qPCR data, more accurate experimental data can be obtained. Herein, the expression stability of alpha-tubulin1 (TUA1), beta-tubulin (TUB), beta-tubulin 1 (TUB1), beta-tubulin 5 (TUB5), actin 1 (ACT1), actin 97 (ACT97), molecular chaperone dnaj (DNAJ), adenine phosphoribosyl transferase (APT), and histone H4 (HIS4) genes from Paulownia fortunei (Seem.) Hemsl. under different experimental conditions (different tissues, drought, salinity, Cd, and Cr treatments) was assessed with four statistical tools: RefFinder, BestKeeper, NormFinder, and geNorm. Notably, TUA1 and TUB5 were identified as CRGs for different tissues, ACT97 and TUB1 for drought treatment, ACT97 and APT for salinity treatment, TUB1 and ACT97 for Cd treatment, and DNAJ, TUB1 and TUB5 for Cr treatment. Furthermore, the results of "total" group, V4/V5 > 0.15 and V5/V6 < 0.15 revealed that the CRGs or gene combinations, which could meet all the test conditions, were not easy to identify. To further verify the reliability of CRGs, the expression levels of paulownia fortunei cellulose synthase A catalytic subunit2 (PfCesA2) and paulownia fortunei glutathione reductase (GR) genes were analysed. The expression patterns were different when the unstable CRGs were used for normalization compared to when the stable CRGs and combination were used for normalization. This study will lay a foundation for study on the expression levels of key genes from P. fortunei seedlings.

Abstract Background Rheumatoid arthritis (RA) is a chronic, systemic autoimmune inflammatory disease, the pathogenesis of which is not clear. Clinical remission, or decreased disease activity, is the aim of treatment for RA. However, our understanding of disease activity is inadequate, and clinical remission rates for RA are generally poor. In this study, we used multi-omics profiling to study potential alterations in rheumatoid arthritis with different disease activity levels. Methods Fecal and plasma samples from 131 rheumatoid arthritis (RA) patients and 50 healthy subjects were collected for 16S rRNA sequencing, internally transcribed spacer (ITS) sequencing, and liquid chromatography-tandem mass spectrometry (LC–MS/MS). The PBMCS were also collected for RNA sequencing and whole exome sequencing (WES). The disease groups, based on 28 joints and ESR (DAS28), were divided into DAS28L, DAS28M, and DAS28H groups. Three random forest models were constructed and verified with an external validation cohort of 93 subjects. Results Our findings revealed significant alterations in plasma metabolites and gut microbiota in RA patients with different disease activities. Moreover, plasma metabolites, especially lipid metabolites, demonstrated a significant correlation with the DAS28 score and also associations with gut bacteria and fungi. KEGG pathway enrichment analysis of plasma metabolites and RNA sequencing data demonstrated alterations in the lipid metabolic pathway in RA progression. Whole exome sequencing (WES) results have shown that non-synonymous single nucleotide variants (nsSNV) of the HLA-DRB1 and HLA-DRB5 gene locus were associated with the disease activity of RA. Furthermore, we developed a disease classifier based on plasma metabolites and gut microbiota that effectively discriminated RA patients with different disease activity in both the discovery cohort and the external validation cohort. Conclusion Overall, our multi-omics analysis confirmed that RA patients with different disease activity were altered in plasma metabolites, gut microbiota composition, transcript levels, and DNA. Our study identified the relationship between gut microbiota and plasma metabolites and RA disease activity, which may provide a novel therapeutic direction for improving the clinical remission rate of RA.