Changhong Xiao

Background: Xinfeng Capsule (XFC) is a traditional Chinese medicine compound preparation that has been clinically used to treat rheumatoid arthritis (RA) for more than 20 years. It has demonstrated clear therapeutic effects, including improving pulmonary function and reducing lung injury in patients with RA. However, the precise mechanism underlying its protective effect against lung injury remains unclear. This study aims to explore the potential mechanisms of XFC in the treatment of lung injury. Methods: Liquid chromatography-mass spectrometry (LC-MS) analysis was conducted to determine the chemical composition of XFC. Proteomic and bioinformatic analyses of differentially expressed proteins (DEPs) in rat lung tissue were performed using tandem mass tag labeling. A rat adjuvant arthritis (AA) model was established using Freund’s complete adjuvant to observe pathological changes in synovial and lung tissues, as well as alterations in lung function. In addition, a cell model was constructed by inducing lung fibroblasts with transforming growth factor-β1 (TGF-β1) to investigate the effects of XFC-containing serum on oxidative stress and pulmonary fibrosis through the peroxisome proliferator-activated receptor gamma (PPARγ)/3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2) pathway. Results: LC-MS analysis identified a total of 867 compounds in XFC, of which 25 unique compounds were closely associated with pulmonary fibrosis and lung injury. Proteomic analysis suggested that XFC may regulate PPAR signaling pathway-related proteins and alleviate lung injury in AA rats. Animal experiments showed that XFC significantly inhibited immune inflammation, synovial hyperplasia, and oxidative stress in AA rats, while reducing lung injury and improving lung function. Furthermore, XFC-containing serum suppressed TGF-β1–induced proliferation of lung fibroblasts, promoted PPARγ expression, and significantly decreased the levels of interleukin-6, tumor necrosis factor-α, reactive oxygen species, nicotinamide adenine dinucleotide phosphate oxidase 4, HMGCS2, collagen type I α 1, collagen type III α 1, and α-smooth muscle actin ( P < 0.01). In addition, XFC partially reversed the effects of the PPARγ antagonist GW9662, activated the PPARγ signaling pathway, inhibited oxidative stress and inflammatory responses, and exerted anti-fibrotic effects similar to those of the PPARγ agonist rosiglitazone. Conclusion: XFC inhibits inflammation and oxidative stress by regulating the PPARγ/HMGCS2 pathway, thereby attenuating fibrosis and alleviating lung injury.

Introduction: Persistent inflammatory refractory rheumatoid arthritis (PIRRA) presents a major clinical challenge, and its underlying molecular mechanisms remain inadequately understood.Methods: athogenesis. Synovial joint tissues were collected from 30 TgTC mice and 30 Friend virus B (FVB) control mice. Of these, 18 mice per group were used for transcriptomic, proteomic, and metabolomic analyses; 6 for pathological examination and microCT imaging; and 6 for validation experiments. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, protein-protein interaction networks, and KEGG Markup Language (KGML) network analysis were employed to characterize the functional roles of differentially expressed genes (DEGs), proteins, metabolites, and associated biological pathways. Notably, five genes/proteins—macrophage-expressed gene 1 (Mpeg1), ectonucleotide pyrophosphatase/phosphodiesterase 2 (Enpp2), toll-like receptor 2 (Tlr2), cluster of differentiation 14 (CD14), and lysozyme 2 (Lyz2)—were validated by quantitative reverse transcription PCR (qRT-PCR), Western blotting, and immunohistochemistry.Results: A total of 2,410 DEGs, 366 differentially expressed proteins, and 120 significantly altered metabolites (P < 0.05) were identified between the model (TgTC ) and control (FVB) groups. These molecules were mainly associated with Golgi apparatus dysfunction, lipid metabolism, and immune-inflammatory responses. Integrative multi-omics analysis further revealed that these molecular alterations are involved in the activation of the PI3K-AKT-mTOR signaling pathway, as well as disruptions in tryptophan and lipid metabolism. Among the metabolites, phosphatidylinositol (PI) (12:0/12:0), N-docosahexaenoyl tryptophan, and PI (22:1(11Z)/0:0) were identified as key metabolic signatures of persistent joint synovitis in TgTC mice. In addition, the expression of Mpeg1, Enpp2, Tlr2, CD14, and Lyz2 was evaluated in synovial samples from patients with PIRRA and classical RA. Notably, Mpeg1, Enpp2, and Lyz2 were significantly upregulated in PIRRA, whereas Tlr2 and CD14 did not show statistically significant differences between groups.Discussion: Our findings highlight the critical role of altered gene, protein, and metabolite expression in the pathogenesis of PIRRA, offering new insights into its molecular basis and potential therapeutic targets.

Background: Persistent synovial hyperplasia with inflammation in rheumatoid arthritis is one of the main pathogeneses of refractory rheumatoid arthritis (RRA). Photodynamic therapy (PDT) causes less trauma than steroid injections or arthroscopic synovectomy while providing stronger targeting and more durable curative effects. The aim of this trial was to evaluate the short-, medium-, and long-term clinical efficacy of PDT when applied as a treatment for RRA synovial hyperplasia and synovitis. Methods and analysis: This protocol is for a single-center, randomized, double-blind, blank-controlled prospective trial. A sample of 126 RRA patients will be randomly divided into 3 groups: the control group, the “PDT once” group, and the “PDT twice” group, with 42 participants per group. The trial will be conducted by the Rheumatology and Immunology Department of the Integrated Hospital of Traditional Chinese Medicine, Southern Medical University. The Ultrasound Compound Score of Synovitis (UCSS) has been selected as the primary outcome measure. The secondary outcome measures include knee joint clinical assessments, ratio of relapse, duration of remission, Disease Activity Score in 28 joints (DAS28), inflammation indexes, serum concentrations of specific antibodies, and changes in articular structures as detected by X-ray scans in the 48th week. The improvement ratios of the UCSS at the 8th, 24th, and 48th weeks (compared with baseline) reflect short-, medium-, and long-term time frames, respectively. Ethics and dissemination: The protocol was approved by the Medical Ethics Committee of the Integrated Hospital of Traditional Chinese Medicine, Southern Medical University, China (Approval No. granted by the ethics committee: NFZXYEC-2017-005) and then entered in the Chinese Clinical Trials Registry under registration number ChiCTR1800014918 (approval date: February 21, 2018). All procedures are in accordance with Chinese laws and regulations and with the Declaration of Helsinki by the World Medical Association (WMA). Any modifications of this protocol during execution will need additional approval from the Ethics Committee of our hospital. Trial registration number: ChiCTR1800014918.

Background Persistent synovial hyperplasia with inflammation in rheumatoid arthritis is the main cause of refractory rheumatoid arthritis (RRA). As a means of local treatment, photodynamic therapy (PDT) confers less trauma, stronger targeting, and more durable curative effects than steroid injections or arthroscopic synovectomy. The aim of this trial will be to evaluate the short-, medium- and long-term clinical efficacy of PDT in the treatment of RRA synovial hyperplasia and synovitis. Methods and analysis This is a single-centre, randomised, double-blind, blank-controlled, prospective trial. A sample of 126 RRA patients will be randomly divided into 3 groups: the control group, the PDT once group, and the PDT twice group, 42 per group. The trial will be conducted at the Rheumatology and Immunology Department of Integrated Hospital of Traditional Chinese Medicine, Southern Medical University. Assessments at baseline, the first operation, the second operation (4th week), and then at three follow-ups (8th week, 24th week, 48th week) will be performed. The Ultrasound Compound Score of Synovitis (UCSS), knee joint clinical assessments, Disease Activity Score in 28 Joints (DAS28), serological inflammation indexes and specific antibody levels, pathological biopsies of synovial tissue and X-ray assessments of bone destruction will be evaluated. An improvement in the UCSS will be the main endpoint, and the UCSS at the 8th week versus the baseline value will reflect the short-term outcome of the operation. The results of the 24th week and 48th week follow-up will reflect the medium- and long-term curative effects, respectively. Ethics and dissemination The protocol was approved by the Medical Ethics Committee of Integrated Hospital of Traditional Chinese Medicine, Southern Medical University, China (Approval No. of the ethics committee: NFZXYEC-2017-005) and later registered in the Chinese Clinical Trials Registry with Registration number ChiCTR1800014918 (approval date: February 21, 2018). All procedures will be in accordance with Chinese laws and regulations, as well as the WMA Declaration of Helsinki. Any modifications to the protocol will be approved by the Ethics Committee of our hospital. Trial registration number ChiCTR1800014918.

Objectives. To determine differences in AIM2 inflammasome expression levels between rheumatoid arthritis (RA) and osteoarthritis (OA) and to investigate the role of AIM2 in RA fibroblast-like synoviocytes (RA-FLS). Methods. Serum AIM2 levels among health controls (HC, ), OA ( ), and RA (n =49) patients were compared via ELISA. The different expression levels of AIM2, ASC, caspase-1, and IL-1β between RA and OA synovium were semiquantified by qRT-PCR and immunohistochemical (IHC) staining. IHC staining was recorded by scores, and its correlation with the ESR and CRP levels of RA patients was determined. SiRNA AIM2 was transferred to RA-FLS and its effects on the proliferation and migration via CCK-8 assay and Transwell test, respectively. Results. In RA sera, the HC expressed higher level of AIM2 than OA and RA patients, and ASC, caspase-1, and IL-1β expressed higher in RA patients than HC; no significant differences were observed between sera of OA and RA patients. However, in affected knee synovium, AIM2, ASC, caspase-1, and IL-1β were expressed higher in RA than that of OA. Moreover, the scores of AIM2, ASC, and IL-1β were positively correlated with the ESR and CRP levels in RA patients. The proliferation of FLS was significantly inhibited after transferring with AIM2 siRNA to FLS. There were no differences in apoptosis and migration assay between the si-AIM2 group and the control group. Conclusion. AIM2 inflammasome pathway involves in the pathogenesis of RA. si-AIM2 inhibits the proliferation of RA-FLS, which may be a promising therapeutic strategy for the treatment of RA.

Objective To explore the relationships of plasma adenylate-activated protein kinase α1 (AMPKα1) and phosphorylated AMPK (p-AMPK) levels of rheumatoid arthritis (RA) with inflammatory indexes. Methods The study recruited 15 health controls (HCs), 17 patients with osteoarthritis (OA) and 61 patients with RA[including 22 cases of very active (VRA), 20 cases of moderate active (MRA) and 19 cases of inactive (IRA) based on the disease activity score in 28 joints-erythrocyte sedimentation rate (DAS28-ESR)]. The levels of ESR and C-reactive protein (CRP) in the patients were measured; the levels of plasma AMPKα1 and p-AMPK as well as interleukin-17 (IL-17) and tumor necrosis factor-α (TNF-α) were detected by ELISA. Pearson r test was used to compare the correlations of AMPKα1 and p-AMPK levels with the DAS28 score, inflammation indexes and inflammatory cytokines. Results Compared with the HCs, the plasma AMPKα1 and p-AMPK levels of the OA patients and RA patients with different disease activities were higher in a certain extent. P-AMPK levels showed a moderately positive correlation with DAS28 score (r=0.27, 95% CI: 0.01-0.49). AMPKα1 showed a moderately positive correlation with TNF-α levels (r=0.46, 95% CI: 0.24-0.64) in the RA patients. Conclusion The p-AMPK level may increase in the plasma of RA patients, and it has a positive correlation with clinical disease activity and inflammation indicators.

Background To determine whether any differences of AIM2 inflammasome expression levels between rheumatoid arthritis (RA) and osteoarthritis (OA) and investigate the effects of AIM2 when transferred into RA fibroblast-like synoviocytes (RA-FLS).Methods Serum AIM2 levels between OA and RA patients were compared by ELISA. Different expression levels of AIM2, ASC, Caspase-1 and IL-1β between RA and OA synovium were semi-quantified by RT-qPCR and immunohistochemical (IHC) staining. IHC staining were recorded by H scores, and determine the correlation with ESR and CRP levels of RA patients. SiRNA AIM2 was transferred to RA-FLS and observe its effects on proliferation and migration by MTT assay and transwell test respectively.Results In RA sera, no significant difference was observed between OA and RA patients. However, in affected knee synovium, AIM2, ASC, Caspase-1 and IL-1β were expressed higher in RA than that of OA. Plus, H score of AIM2, ASC, and IL-1β were positively correlated to ESR and CRP levels in RA patients. After transferred AIM2 siRNA to FLS and incubation for 48 hours, the proliferation of FLS were significantly inhibited, and the apoptosis rate were significantly increased compared to FLS in control group. However, no effect on migration was detected.Conclusions AIM2 participated in the proliferation of FLS, and might be a potential target for therapy.