The formation of chronic refractory wound (CRW) is the result of the combined effect of multiple factors, among which fibrosis plays a dual role in the pathogenesis. Moderate fibrosis can protect the wound, whereas excessive fibrosis leads to pathological scarring, which aggravates tissue hypoxia and stiffness, thereby impeding healing. Thymic stromal lymphopoietin (TSLP) is an important immunoregulatory cytokine that plays a critical role in allergic diseases. Recent studies have revealed that TSLP is also involved in the fibrotic mechanisms underlying CRWs. TSLP regulates key signaling pathways, including transforming growth factor-beta/Sma- and Mad-related protein (TGF-β/SMAD), wingless-type mice mammary tumour virus integration site family/beta-catenin (Wnt/β-catenin), and phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT), thereby affecting fibroblast activation, macrophage polarization, chemokine expression, and lysyl oxidase-mediated collagen cross-linking, ultimately contributing to CRW fibrosis. Future studies should focus on exploring the synergistic effects of TSLP with other profibrotic factors and developing combination therapies to balance fibrosis and tissue repair, thereby providing new strategies for the treatment of CRW fibrosis.
Objective To establish a simple and cost-effective protocol for the directed differentiation of induced pluripotent stem cells (iPSCs) into macrophages (Mϕ). Methods Human umbilical cord blood mononuclear cells were reprogrammed into iPSCs by introducing reprogramming factors with Sendai virus vectors. The iPSCs were then induced to differentiate into Mϕ through an embryoid body-based approach, combined with key cytokines including bone morphogenetic protein 4 (BMP4), macrophage colony-stimulating factor (M-CSF), and interleukin-3 (IL-3). Flow cytometry, real-time quantitative polymerase chain reaction and other methods were used to determine the phenotype, function, and expression of immunogenicity-related molecules of the differentiated Mϕ. Results High-purity CD14+CD11b+ Mϕ with normal phagocytic function and polarization capacity were successfully generated by this protocol, without co-culture with animal-derived feeder cells, hypoxic conditions, or cell sorting at intermediate stages. Immunophenotypic analysis revealed that the immune checkpoint CD276+ cells were reduced after the differentiation of iPSCs into Mϕ, which was inconsistent with the stable expression of CD276 observed during the differentiation of iPSCs into β cells or retinal pigment epithelial cells. Conclusion In this study, high-purity Mϕ were successfully generated using a feeder-free, hypoxia-free, and sorting-free system. Furthermore, the lineage-dependent downregulation of CD276 during Mϕ differentiation was observed for the first time.
Objective To establish a method for isolating and expanding perivascular adipose-derived mesenchymal stem cells (PV-ADSC) from rat aortas and to provide a reliable cellular model for investigating vascular remodeling and the related cardiovascular diseases. Methods Perivascular adipose tissue (PVAT) was aseptically collected from the aortas of 1-month-old Sprague Dawley (SD) rats, then rinsed, minced, digested, filtered, and seeded for culture. Upon reaching 80%-90% confluence, the cells were passaged for expansion, and their morphology and proliferation were continuously monitored. Passage 5 (P5) cells were harvested for flow cytometric analysis of surface CD marker expression, and directed differentiation assays with lineage-specific staining were performed to assess the trilineage (adipogenic, osteogenic, chondrogenic) differentiation potential. Results On day 4 of the primary culture, spindle-shaped, stellate, and polygonal cells were observed migrating from the explant edges. Between day 5 and 6, island-like cell clones emerged, and by day 7 to 8, the colonies had coalesced into a confluent monolayer with swirling or parallel alignment. After being passaged, the cells exhibited a homogeneous fibroblast-like spindle morphology and maintained robust proliferation, with no evident senescence or aberrant differentiation through P8. P5 cells showed high expression of the mesenchymal stem cells (MSC) surface markers CD90 (93.60±0.62)%, CD73 (86.80±2.40)%, CD44 (97.77±0.58)%, and CD29 (98.60±0.30)%, yielding a mean positivity rate of (94.19±5.00)%. In contrast, expressions of the hematopoietic stem cells (HSC) surface markers CD45 (5.42±0.78)%, CD34 (7.15±0.49)%, and CD11b/c (3.74±0.08)% were significantly lower, with a mean rate of only (5.44±1.55)%. Directed differentiation assays confirmed the cells' ability to differentiate into adipocytes, osteoblasts, and chondrocytes, as demonstrated by positive staining with Oil Red O, Alizarin Red, and Alcian Blue, respectively. Conclusion Using the method established in this study, rat aortic PV-ADSC were successfully isolated and expanded. These cells exhibited the morphological and surface marker profiles typical of MSC, possessed robust self-renewal capacity, and retained the potential for trilineage differentiation into adipogenic, osteogenic, and chondrogenic lineages. Collectively, these characteristics establish a reliable cellular model for studying vascular remodeling and the related cardiovascular diseases.
B-cell epitopes are regions on the surface of antigens that can be recognized and bound by B-cell receptors or antibodies. Accurate identification of B-cell epitopes of antigens can accelerate the development of drugs such as antibodies and vaccines. This paper conducts a systematic review of representative methods developed in recent years for predicting both B-cell conformational epitopes (such as SEPPA 3.0, DiscoTope-3.0, BepiPred-3.0, etc.) and linear epitopes (such as DLBEpitope, EpiDope, EpitopeVec, etc.). It presents an introduction to these methods, covering aspects such as model architecture, prediction performance, application scope, and illustrative examples, with the aim of providing reference information for related research fields.
Objective To investigate the role of berberine (BBR) in chronic lymphocytic leukemia (CLL) and to determine whether it exerts anti-tumor effects by directly targeting and inhibiting Lck/Yes tyrosine kinase (Lyn), a novel Src family kinase, thereby inducing leukemia cell apoptosis. Methods Molecular docking was employed to predict the binding potential between BBR and Lyn kinase, and biotin pull-down assay was conducted to validate their direct interaction. In vitro experiments utilized the human chronic B-cell leukemia cell line MEC-1, with cell viability and apoptosis assessed via CCK-8 and TUNEL staining, respectively. Key proteins in the B-cell receptor (BCR) pathway, including Lyn, spleen tyrosine kinase (Syk), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), and apoptosis-related markers Bcl2-associated X protein (BAX), Bcl2-associated agonist of cell death (BAD), cleaved caspase-3(c-caspase-3), and B-cell lymphoma 2 (Bcl2), were analyzed by Western blot. Transcriptional levels of downstream genes, including cyclin D1 (Cyclin D1), Bcl2, and myelocytomatosis viral oncogene homolog (c-Myc), were quantified using real-time quantitative PCR. Functional rescue experiments were performed using Lyn-overexpressing lentiviral stable cell lines. In vivo, a C-NKG mouse leukemia model was established via tail vein injection, with tumor infiltration in the spleen, liver, and lungs evaluated by HE staining, and therapeutic effect of BBR assessed by survival analysis. Results In vitro, BBR inhibited MEC-1 cell proliferation in a concentration-dependent manner and induced apoptosis, while suppressing the phosphorylation of BCR pathway proteins and downstream gene expression. Molecular docking and pull-down assays confirmed the direct binding between BBR and Lyn. The overexpression of Lyn reversed BBR-induced apoptosis and pathway inhibition. In vivo, BBR treatment significantly reduced organ infiltration and prolonged survival in leukemic mice, which can be reversed by Lyn overexpression. Conclusion BBR induces CLL cell apoptosis and inhibits tumor progression in vitro and in vivo by directly targeting Lyn kinase and suppressing the BCR-Lyn-PI3K-AKT signaling pathway. These findings provide experimental evidences supporting BBR as a natural Lyn-targeted therapeutic agent for CLL.
Type 3 innate lymphoid cells (ILC3s) are a subset of innate immune cells regulated by the transcription factor retinoic acid receptor-related orphan receptor γt (RORγt). They are primarily distributed in the intestinal mucosa and lymphoid tissues, where they secrete cytokines such as interleukin 17(IL-17), IL-22, and granulocyte-macrophage colony-stimulating factor (GM-CSF), playing a crucial role in maintaining intestinal mucosal homeostasis, defending against pathogen invasion, and modulating immune responses. Renal fibrosis (RF) is a pathological process triggered by pathogenic factors such as trauma, infection, inflammation, or metabolic abnormalities. It is characterized by damage to renal parenchymal cells, abnormal deposition of extracellular matrix (ECM), and progressive fibrosis of renal tissue, ultimately leading to irreversible loss of kidney function. Recent studies have revealed that ILC3s not only contribute to intestinal homeostasis but are also involved in the progression of RF. Therefore, this review summarizes the research advances in the mechanisms of ILC3-mediated cross-organ regulation in RF, aiming to provide new perspectives for the prevention and treatment of RF.
Objective To investigate the effects of Cinobufotalin (Cino) on the tumor microenvironment (TME) of colorectal cancer (CRC) and the chemosensitivity to Oxaliplatin (OX). Methods We explored the clinical expression differences of CD244 in the TCGA-COAD cohort. Through UMAP dimensionality reduction analysis of single-cell sequencing data, we clarified the cellular localization of CD244 and analyzed its interaction characteristics with immune cells. In vivo experiments, a CRC tumor-bearing mouse model was established and randomly divided into 4 groups: Tu group, Cino group, OX group, and Cino combined with OX group. Intraperitoneal injection was performed every other day for treatment. Experimental techniques such as tumor volume measurement, fluorescence signal/mass changes and tissue morphology changes, real-time quantitative PCR, Western blot, and immunofluorescence staining were used to detect tumor proliferation, metastasis, expression distribution of CD244+ macrophages and T cell activity. In vitro experiments, RAW264.7 cells were stimulated with interleukin 4 (IL-4) to construct a M2-type macrophage model. The effects of Cino, OX or Cino combined with OX on the expression of CD244+ macrophages were observed, and the conditioned medium (CM) of the corresponding treated macrophages was used to culture T cells. The expression of T cell exhaustion and activation markers was detected, and the killing function of T cells was tested. Results In vivo experiments, Cino treatment had no significant effect on tumor growth, proliferation and metastasis in mice. OX treatment and its combination with Cino inhibited tumor proliferation and metastasis, with the Cino-OX combination exhibiting an enhanced inhibitory effect. Both Cino and OX inhibited the expression of CD244+ macrophages in tumor tissues, and their combination showed a synergistic inhibitory effect. Neither Cino nor OX alone had significant effect on T cell activity, their combination could inhibit T cell exhaustion and increase CD8+ T cell infiltration. In vitro experiments, Cino and OX treatments inhibited the expression of CD244+ macrophages, with their combination exhibiting an enhanced inhibitory effect; after IL-4 treatment, the supernatant was collected to culture T cells, and the tumor-killing ability of T cells decreased. The CM treated with Cino or OX could enhance the killing ability of T cells, and the combination showed a stronger killing ability. Conclusion Cino enhances the tumor-killing capacity of T cells and reshapes the tumor microenvironment by specifically targeting and inhibiting CD244+ macrophages, thereby increasing the chemosensitivity of CRC to OX.
Objective To investigate the distribution characteristics of the genotypes and allele frequencies of HNA-1~-5 alleles in a blood donor population from Jiangsu Province. Methods Genomic DNA was extracted from 300 peripheral blood samples collected from healthy donors. All samples were genotyped using an optimized polymerase chain reaction-sequence specific primer (PCR-SSP) protocol. Randomly selected results were verified by direct sequencing, and genotype distributions were tested for conformity to Hardy-Weinberg equilibrium. Samples identified as potential HNA-1 null were confirmed by polymerase chain reaction sequencing-based typing (PCR-SBT) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results The allele frequencies for HNA-1a, HNA-1b, and HNA-1c were 58.6%, 41.4%, and 0%, respectively. Two identified HNA-1 null cases were confirmed to result from a complete deletion of the Fc gamma receptor IIIb (FCGR3B) gene. The allele frequency of the HNA-2-encoding CD177 gene was 100%, with no HNA-2 null cases detected due to the homozygous c.787 A>T (TT) mutation. The allele frequencies for HNA-3a, HNA-3b, HNA-4a, HNA-4b, HNA-5a, and HNA-5b were 63.0%, 37.0%, 100%, 0%, 88.8%, and 11.2%, respectively. No statistically significant differences in HNA allele frequencies were found between the Jiangsu population and those from Shanghai or Zhejiang. However, significant differences were observed when compared to the Guangzhou population specifically for the HNA-1a/1a (P=0.008), HNA-3a/3a (P=0.033), and HNA-3b/3b (P=0.022) genotypes. Conclusion This study establishes the HNA genotype and allele frequency profile for the Jiangsu blood donor population, providing essential baseline data for research into HNA-related alloimmunization. Furthermore, it optimizes a methodology suitable for large-scale HNA screening and establishes a comprehensive confirmatory testing protocol for HNA-1 null, thereby laying a technical foundation for subsequent research on this allele.
Human cytomegalovirus (HCMV) infection is one of the most common and severe viral complications following allogeneic hematopoietic stem cell transplantation (allo-HSCT), significantly increasing the risk of graft-versus-host disease (GVHD) and non-relapse mortality. Letermovir, as the first inhibitor targeting the HCMV terminase complex, blocks the cleavage of viral DNA and its packaging into viral capsids, demonstrating remarkable efficacy in early post-transplant HCMV prophylaxis. It significantly reduces the incidence of clinically significant HCMV infection (csCMVi) and indirectly contributes to improved GVHD- and relapse-free survival. However, while the potent viral suppression of this treatment provides clinical benefits, it also limits viral antigen exposure, which leads to delays in the reconstitution of HCMV-specific T cells and NK cells and consquently the increased risk of late-onset HCMV reactivation after drug discontinuation. This may also be associated with an elevated risk of Epstein-Barr virus reactivation and post-transplant lymphoproliferative disease (PTLD). This review aims to systematically summarize the dual effects of letermovir in allo-HSCT and analyzes its pharmacological mechanisms, clinical efficacy, impact on immune reconstitution, and potential consequences. Furthermore, it explores individualized management strategies based on immune monitoring to provide a theoretical basis for optimizing clinical practice.
Objective To explore the immune landscape of patients with lung adenocarcinoma with drainage lymph nodes by single-cell sequencing technology. Methods The lung cancer single cell dataset (GSE277742) was obtained from the Gene Expression Omnibus (GEO) database of the National Center for Biotechnology Information (NCBI). The dataset included intrathoracic DLN from 18 patients with pathologically confirmed lymph node metastasis and 4 control patients without evidence of metastasis. R language was used for data quality control, dimensionality reduction clustering, cell subgroup annotation and cell communication analysis to identify key cell subsets and screen differentially expressed genes. Kaplan-Meier survival analysis was performed using the TCGA database. Results Single cell sequencing analysis showed that the proportion of CD8+ T cells in patients with draining lymph nodes was significantly higher, among which cyclin-dependent kinase 6(CDK6), cyclin D3(CCND3), CCNH, cyclin-dependent kinase inhibitor 2A/B/D(CDKN2A/B/D) and retinoblastoma transcriptional corepressor 1(RB1) were highly expressed. TCGA database analysis showed that the expression of genes related to the BIOCARTA_CELLCYCLE_PATHWAY pathway was significantly different between tumor and normal samples, and was related to survival. Conclusion Several key genes differentially expressed in the draining lymph nodes of patients with lung adenocarcinoma are screened by bioinformatics methods, which provides a new perspective for further analysis of the regulatory mechanism of the immune microenvironment of lung adenocarcinoma with draining lymph nodes.
Malignant tumors are major diseases that threaten human health. A long-standing focus of research is how to eliminate tumor cells while minimizing damage to healthy tissues. This article systematically reviews the dual role of iron metabolism in tumorigenesis and progression. Firstly, disruption of iron homeostasis promotes malignant transformation through mechanisms including oxidative stress, signaling transduction, and epigenetic regulation. Secondly, ferroptosis triggered by iron overload offers a novel avenue for targeted cancer therapy. Based on the established relationship between iron metabolism and cancer progression, this paper proposes therapeutic strategies centered on iron chelators, ferroptosis inducers, and nanomaterials. It further elaborates on recent advances in combining iron metabolism modulation with chemotherapy, immunotherapy, epigenetic therapy, and other modalities. Finally, by elucidating the core mechanism through which iron metabolism promotes cancer, we suggest potential therapeutic directions, aiming to provide new insights for advancing precision oncology and developing novel anticancer agents.
The progression of atherosclerosis (AS) is intrinsically associated with dysregulated lipid metabolism in macrophages, leading to the formation of foam cells. MicroRNA (miRNA) finely regulates the expression of key genes that govern cholesterol uptake, esterification, and efflux, thereby maintaining lipid homeostasis. Aberrant miRNA expression disrupts this balance and accelerates lipid accumulation. In recent years, the potential of miRNA as a biomarker for early diagnosis and as a target for therapeutic intervention in AS has gained considerable attention. Although miRNA-based strategies have shown encouraging results in preclinical studies, their clinical translation is still limited by inadequate delivery efficiency and off-target effects. Future research should prioritize the development of efficient and highly specific delivery platforms, as well as the integration of miRNA-based therapies with other treatment modalities, to pave the way for precision management of atherosclerosis.
Objective This study aimed to investigate the impact of human leukocyte antigen B27 (HLA-B27)/β2m gene expression on the gut microbiota and metabolites, and to elucidate its role in the pathogenesis of spinal arthritis (SpA)-associated intestinal inflammation. Methods Transgenic mice expressing HLA-B27/β2m without spontaneous inflammation were employed. Integrated multi-omics analyses, including metagenomics and metabolomics, were conducted to profile microbial and metabolic changes at prenatal, early colonization, and stable colonization stages. Inflammatory susceptibility was further assessed using a dextran sulfate sodium (DSS)-induced colitis model. Results Expression of HLA-B27/β2m significantly altered the gut microbiota structure, promoting the expansion of Gram-negative bacteria and inhibiting Gram-positive populations. Metabolomic profiling revealed enhanced arachidonic acid metabolism, elevated levels of pro-inflammatory metabolites such as prostaglandins, and a reduction in anti-inflammatory flavonoids. These findings collectively indicated a pro-inflammatory intestinal microenvironment, which was corroborated by exacerbated colitis upon DSS challenge in animal models. Conclusion The HLA-B27/β2m gene modulates gut microbial composition and metabolic balance, predisposing the intestine to inflammatory responses. These results provide novel mechanistic insights into the "gut-joint axis" in SpA pathogenesis.
Objective To investigate the molecular mechanisms by which long-stranded non-coding RNA (lncRNA) differentiated antagonistic non-protein-coding RNA (DANCR) regulates ubiquitin-specific protease 33 (USP33) and the glutathione peroxidase 4 (GPX4) in the context of Mycobacterium tuberculosis (Mtb) H37Ra infection. Methods A DANCR overexpression model was established using the THP-1 cell line. By integrating bioinformatics prediction, real-time quantitative PCR, Western blot, RNA immunoprecipitation, co-immunoprecipitation-proteomics, and molecular docking techniques, the regulatory effects of DANCR on miRNA, USP33, and GPX4 were systematically analyzed, as well as the biological functions of DANCR under the infection of the attenuated Mtb strain H37Ra. Results DANCR acted as a competitive endogenous RNA (ceRNA), specifically sponging miR-19a-3p, and relieving the post-transcriptional inhibition of miR-19a-3p on USP33, thereby significantly up-regulating the mRNA and protein levels of USP33. Molecular docking and co-immunoprecipitation experiments suggested that USP33 and GPX4 interacted physically and overexpression of DANCR induced the protein level of GPX4 without affecting its mRNA, indicating that this regulation occurs at the post-translational level. Conclusion The study suggested a key DANCR-miR-19a-3p-USP33 regulatory axis, in which the long non-coding RNA DANCR acts as a ceRNA by sponging miR-19a-3p to relieve its inhibition on USP33, thereby upregulating USP33 expression. Meanwhile, the results of bioinformatics analysis and co-immunoprecipitation indicated that USP33 has a direct physical interaction with the ferroptosis key protein GPX4, and it is speculated that USP33 may stabilize GPX4 through deubiquitination modification, providing new molecular clues and therapeutic targets for understanding the metabolic regulation, oxidative stress response, and potential ferroptosis mechanism in the interaction between the host and Mtb.
Chimeric antigen receptor natural killer (CAR-NK) cell therapy has shown broad prospects in the field of tumor immunotherapy, owing to its advantages such as low immunogenicity, "off-the-shelf" preparation and a good safety record. However, in the treatment of solid tumors, it faces core challenges including insufficient in vivo persistence, low homing efficiency, and immunosuppression within the tumor microenvironment (TME). Umbilical cord mesenchymal stem cells (UC-MSCs) have unique abilities in immune regulation, paracrine secretion, and remodeling of the TME, providing a new strategy for overcoming the limitations of CAR-NK cell therapy. This review summarizes the recent research progress on both CAR-NK cells and UC-MSCs in tumor immunotherapy. It further discusses the synergistic mechanism and experimental evidence supporting the combination of UC-MSCs with CAR-NK cells, and analyzes the challenges and development prospects of this method as a new tumor immunotherapy.
Objective To investigate the expression characteristics of the interleukin 2-inducible T-cell kinase (ITK) gene in lung adenocarcinoma (LUAD), its clinical prognostic value, and its potential role in the tumor microenvironment. Methods ITK expression data were obtained from the TCGA and GEO datasets (GSE75037, GSE32863) to analyze its differential expressions between LUAD and adjacent normal tissues. The TCGA-LUAD cohort was used for survival analysis, clinicopathological correlation analysis, and to construct an integrated nomogram prognostic model combining ITK expression with clinical features via Cox regression. The TIMER and CIBERSORT algorithms were employed to assess the correlation between ITK expression and immune cell infiltration. GSEA enrichment analysis was used to compare pathway differences between the high and low ITK expression groups. Results ITK expression was significantly lowered in multiple cancers including LUAD, and its low expression was associated with poor prognosis of patients. Clinical analysis revealed that ITK expression was significantly correlated with tumor Stage grade and T stage. The constructed nomogram model demonstrated good predictive performance (C-index=0.67). High ITK expression showed a significant positive correlation with increased infiltration of anti-tumor immune cells such as CD8+ T cells and M1 macrophages. Pathway enrichment analysis indicated that the high ITK expression group was primarily enriched in immune-related pathways like inflammatory response and interferon-gamma signaling, whereas the low expression group was enriched in metabolic reprogramming processes such as fatty acid metabolism and glycolysis. Conclusion ITK is a potential independent prognostic biomarker in LUAD. Its loss of expression may be associated with suppressed tumor immune microenvironment and activated metabolic reprogramming, thereby promoting tumor progression. ITK may serve as a novel target for prognostic assessment and immunotherapy research in LUAD.
Tertiary lymphoid structures (TLSs) are key regulatory components of the breast cancer immune microenvironment. Their microscopic composition (specific immune cell subsets, spatial organization) and functional status (maturity) directly determine their performance in coordinating the anti-tumor immune responses, serving as the microscopic foundation for understanding the efficacy of immunotherapy. However, the high heterogeneity and dynamic evolution of the tumor microenvironment pose significant limitations for immunotherapeutic efficacy prediction models based on traditional histopathological staging and molecular subtyping. In this context, developing non-invasive assessment technologies capable of quantitatively analyzing the spatial distribution and functional activity of TLSs holds crucial clinical value for achieving precise immunotherapy response prediction and prognostic stratification. Artificial intelligence (AI) technology, particularly the deep learning algorithms integrating multi-omics data, offers innovative tools for systematically decoding TLSs by leveraging its unique advantages in complex feature extraction and high-dimensional data analysis. This review focuses on how AI technology deciphers the microcosmic nature of TLSs. It systematically summarizes recent advances in AI-driven analysis of multi-modal data, including genomics, pathological images, and medical imaging, to decode TLSs. It also delves into the challenges lying in these technologies (such as data standardization and model interpretability) and envisions future pathways for advancing TLS research from microscopic insights to personalized precision immunotherapy.
Bone metabolism is a dynamic physiological process that relies on the precise balance between bone formation and resorption to maintain bone homeostasis, in which innate immune cells play an indispensable role. Innate immune cells interact directly or indirectly with bone cells, participating in the regulation of inflammatory and immune processes and affecting bone metabolism balance. This article aims to clarify the mechanisms by which innate immune cells regulate bone metabolism, thus providing new insights for the prevention and treatment of bone metabolic disorders.
Objective To investigate the mechanism by which Ankle-Three-Needle (ATN) acupuncture alleviates radicular neuropathic pain (RNP) through modulation of pain thresholds and inflammatory responses via 5-hydroxytryptamine (5-HT) signaling. Methods Male Sprague-Dawley (SD) rats were randomly assigned to four groups: sham-operated group, model group, ATN group, and conventional acupuncture group. A rat model of radicular neuropathic pain was established, and corresponding treatments were administered according to group allocation. Pain threshold alterations were assessed using the acetone test, hot plate test, and von Frey filament test. The levels of 5-hydroxytryptamine (5-HT), β-endorphin (β-EP), and substance P (SP) in serum and brain tissues were quantified by enzyme-linked immunosorbent assay (ELISA). Macrophage polarization at the site of sciatic nerve injury and the expression of inflammatory cytokines were analyzed using quantitative real-time PCR (qRT-PCR) and flow cytometry. Rescue experiments involving activation of the 5-HT3 receptor were conducted to elucidate the role of 5-HT signaling in ATN-mediated modulation of pain thresholds and inflammatory responses. Results ATN treatment significantly increased pain thresholds in rats with radicular neuropathic pain, alleviated pain-related behaviors, and inhibited M1 macrophage polarization as well as pro-inflammatory responses at the injury site. Neurochemical analyses demonstrated that ATN upregulated 5-HT levels in the brain while reducing serum 5-HT concentrations. Rescue experiments further confirmed that both 5-HT and the 5-HT3 receptor are essential mediators of ATN-induced regulation of pain thresholds and inflammatory cytokine expression. Conclusion Ankle-Three-Needle acupuncture effectively alleviates radicular neuropathic pain by decreasing serum 5-HT levels and modulating 5-HT3 receptor-mediated macrophage M1/M2 polarization and inflammatory cytokine expression.
Recurrent spontaneous abortion (RSA) is a common pregnancy complication in women of reproductive age. Immune homeostasis at the maternal-fetal interface, maintained by various immune cells and cytokines, is crucial for normal pregnancy. Aberrant immune cell crosstalk disrupts this balance, potentially leading to adverse pregnancy outcomes. This review summarizes the mechanisms of RSA induced by key immune cells and recent advances in dysregulated immune cell interactions at the maternal-fetal interface.
