Introduction. Osteoporosis (OP) is characterized by microstructural degeneration of bone tissue, low bone mass, bone fragility and even brittle fracture (osteoporotic fracture, OPF). OP and OPF are common and there are many disadvantages to the current medications for OP/OPF. Osteoking is a traditional Chinese medicine (TCM) originating from the Yi nationality (Yunnan, China) that has been used to treat bone diseases for decades. Hypothesis/Gap Statement. This study will reveal the changes in the intestinal microbiota of OP rats after 70 days of osteoking treatment. Method. With duplication of sham and OP rats, eight groups were established, with six rats in each group. The intestinal microbiotas were analysed by 16S rDNA sequencing. Results. The results showed that osteoking changed the intestinal microbiota of sham rats and OP rats. The mechanism by which osteoking improves OP is related to the functions of the intestinal microbiota. After 70 days of treatment with osteoking, the contents of Pseudonocardia, Pedomicrobium, Variovorax, Niastella and Actinosynnema were decreased in OP rats. The functions of the above intestinal microbiota related to iron metabolism affected calcifediol and 25(OH)D, and measuring these bone metabolic indicators is required for further study. Conclusion. Osteoking changes the intestinal microbiota to improve OP, and further study which reveals these intestinal microbiota and mechanism is needed.

Background: Primary biliary cholangitis (PBC) is a chronic autoimmune liver disease characterized by cholestasis caused by intrahepatic small bile duct injury. Promoting the molecular mechanism of OTU deubiquitinase 5 (OTUD5) in the treatment of PBC requires further exploration. This study unraveled the molecular underpinnings of PBC through bio-informatics analysis and experimental verification for the development of targeted therapeutic strategies. Methods: This study screened immune-related genes and validated their expression patterns in whole blood of patients with PBC using microarray based on GEO datasets. The expression level of OTU deubiquitinase 5 (OTUD5) was validated in peripheral blood samples using RT-qPCR and immunofluorescence. Subsequently, proteomic bioinformatics analyses were conducted utilizing STRING and InBio Discover databases to predict interactions with the mitochondrial antiviral signalling protein (MAVS). Furthermore, immunochemical and immunofluorescence analyses of MAVS expression in liver tissues were conducted with a thorough analysis of immune cell infiltration specific to the disease by utilizing single-cell RNA sequencing (scRNA-seq) technology in peripheral blood mononuclear cells (PBMCs) derived from both patients with PBC and healthy controls. Results: Compared with those of healthy controls, the liver tissues of patients with PBC presented increased NK cell activation, monocyte/mast cell numbers, and eosinophil numbers. Compared with those in 10 healthy controls, the expression of OTUD5 and MAVS was increased in 16 tissues of patients with PBC. High expression of OTUD5-MAVS in subpopulation 11 mononuclear macrophages was screened by PBMC scRNA-seq, and mononuclear cells with the subgroup 11 phenotype presented highly differentiated characteristics. The expression of OTUD5 and MAVS was inhibited in RAW264.7 cells when OTUD5 was knocked down (P < 0.05). Conclusion: This study focused on the overexpression of OTUD5 and its interaction with MAVS within macrophage subset 11 in patients with primary biliary cholangitis (PBC). The expression of OTUD5 and MAVS is increased in patients with PBC and is a potential target for the diagnosis and treatment of PBC.

Background Knee osteoarthritis (KOA) is a common degenerative bone disease, and transcutaneous electrical nerve stimulation (TENS) is an alternative and complementary therapy (ACM). This study revealed the role of TENS in regulating the intestinal microbiota in KOA rats. Methods This study concentrated on the intestinal microbiota of KOA rats which were treated with TENS for 1, 2, and 3 weeks. Three intensities of TENS were used to treat KOA rats, and the expressions of IL6/8, PI3K-AKT were measured. The intestinal microbiota was analyzed by 16 S rDNA sequencing. Results Compared with the Model Control group, TENS could improve symptoms of KOA rats and inhibit the expressions of IL6/8 by down-regulating the PI3K-AKT expression. After 3 weeks of treatment with TENS, compared with the Model Control group, the abundances of Bacteroidetes, Bacteroidetes, and Thermodesulfobacteria increased in the TENS groups; the abundances of Ficmicutes, Campylobacter, and Verruca decreased in the TENS groups. Conclusion TENS could improve the histomorphology of knee and inhibit inflammation in KOA rats. After treatment with TENS, the intestinal microbiota gradually changed from 1,2,3 weeks and the abundance of them was different with three intensities of TENS. Further study will elucidate the underlying mechanisms of TENS in altering gut microbiota and the potential therapeutic applications of these intestinal microbiota for KOA.

Objective This study revealed the function and mechanism of transcutaneous electrical nerve stimulation (TENS) in a rat model of lateral ankle sprain (LAS). Methods The LAS rat model was randomly divided into Low/Medium/High intensity of TENS groups, Sham group, and Model control group. After 1, 2, and 3 weeks of treatment, the improvement of LAS rar was measured by gait analysis, morphology, micro-CT, osteogenesis markers, etc. The expressions of IL-1β/CXCL-10/NF-kB of ankle joint were analyzed by PCR, IF, IHC and WB. Furthermore, comprehensive profiling of differential gene expression in the LAS rat model was conducted through transcriptome sequencing. Following intra-articular reinfusion of chondrocytes induced by TENS and the downregulation of CXCL-10 expression, the improvements were analyzed. Results After TENS treatment, compared with the model control group, the ankle degeneration score increased in the medium/high intensity of TENS groups (P < 0.05). The degree of inclined plane test increased in the low intensity of TENS group (P < 0.05). The average step length, step frequency increased in the TENS group (P < 0.05). The bone mineral density (BMD)/ bone mineral content (BMC) increased in the TENS groups. The bio-mechanical indicators of LAS rats increased in the TENS groups (P < 0.05). The cartilage thickness of the LAS rat increased in the TENS groups (P < 0.05). There were 7 gene targets of LAS rat that were closely related to the IL-17 signaling pathway; TENS down-regulated the CXCL-10/IL-1β/NF-kB expressions (P < 0.0). After chondrocyte reinfusion which induced with TENS, similar improvements in LAS rats were obtained. Conclusion TENS could improve the motor function and promote the osteogenic and chondrogenic effects in LAS rats. TENS inhibited the inflammation by regulating the IL-1β/CXCL-10/NF-kB expression. The chondrocyte reinfusion with low-expressed CXCL-10, which was induced by TENS, improved the LAS rat. TENS, chondrocyte reinfusion and CXCL-10 inhibitor had a potential effect on LAS in the clinic. Graphical abstract

Background Trefoil factor 3 (TFF3), a naturally occurring cysteine-rich tripeptide in mammals, is predominantly expressed in the goblet cells of the small intestine and colon. As a member of the neuropeptide family, TFF3 has been implicated in a broad spectrum of physiological and pathological processes. Extensive research has demonstrated its involvement in the etiology, progression, and prognosis of eight major categories of human diseases, including those affecting the nervous, digestive, and respiratory systems, with expression levels being modulated in response to different pathological conditions. Purpose This review aims to provide a comprehensive overview of TFF3’s pathological functions across various human diseases, with a particular focus on neurological and psychiatric disorders. We also investigate the associated proteins and signaling pathways that mediate TFF3’s biological activities, in order to establish a theoretical basis for its potential clinical application and identify it as a promising therapeutic target for neuropsychiatric diseases. Results TFF3 has been shown to play a critical role in the pathogenesis of several disease conditions, particularly in the central nervous system. Its capacity to interact with specific receptors and modulate signaling pathways underscores its potential as a key molecule in the development of novel therapeutic strategies. Recent studies on TFF3’s mechanisms of action in neurological disorders highlight its importance as a drug target, with evidence suggesting its involvement in neuroprotection, inflammation, and neuronal repair. These findings contribute to a growing body of literature that positions TFF3 as a versatile and multifunctional peptide with significant therapeutic potential. Conclusion This review highlights the significance of TFF3 as a therapeutic target and underscores the need for further investigation into its molecular mechanisms to facilitate its application in clinical settings.

Background: This study revealed the function and mechanism of the intestinal microbiota in knee osteoarthritis (KOA) rats treated using transcutaneous electrical nerve stimulation (TENS). Methods: KOA model rats were randomly divided into low−/medium−/high‐intensity TENS groups, sodium hyaluronate (SH)‐positive control group (SH), and model control group (KOA rat). After 1, 2, and 3 weeks of treatment, the improvement in KOA severity was assessed, and the expression of interleukin‐1β (IL‐1β)/IL‐6/IL‐8/bone morphogenetic protein 2 (BMP‐2)/transforming growth factor β (TGF‐β) was analyzed. The diversity of the intestinal microbiota in KOA rats was analyzed via 16S ribosomal DNA (rDNA) sequencing. After fecal microbiota transplantation (FMT), which was induced by TENS, the improvement in the intestinal microbiota in KOA rats was analyzed. Results: After 3 weeks of treatment using TENS, compared to those in the model control group, the biomechanical parameters increased in the SH and TENS groups ( p < 0.05); the gait parameters improved in the SH and TENS groups; the bone mineral density (BMD) increased in the TENS group ( p < 0.05); the Mankin scores of the distal femur and proximal tibiofibular muscles decreased in the SH and TENS groups ( p < 0.05). IL‐1β/6/8 expression levels decreased in the SH and TENS groups ( p < 0.05). BMP‐2/TGF‐β expression in the distal femur increased in the TENS group ( p < 0.05). 16S rDNA sequencing revealed that the intestinal microbiota of KOA rats was changed after TENS treatment, including increases in Escherichia‐Shigella , Lachnospira , Eubacterium , Gastranaerophilales , and Rikenellaceae RC9 and decreases in Fusicatenibacter and Mycoplasma . After FMT, which is induced by TENS, similar improvements in KOA rats were obtained. Conclusions: TENS promoted anti‐inflammatory and osteogenic effects by downregulating the Il‐1β/6/8 expression levels and upregulating the BMP‐2/TGF‐β signaling pathway. 16S rDNA sequencing revealed that the intestinal microbiota of KOA rats was changed after TENS treatment via the gut–knee joint axis, and that these dominant genera of FMT elicited improvements in KOA rats. TENS caused improvements in KOA rats by regulating the intestinal microbiota; thus, TENS and induced FMT altered intestinal microbiota suggest a potential novel therapeutic avenue for KOA in clinical settings.