Cervical cancer (CC) is one of the most frequent female malignancies worldwide. However, the molecular mechanism of lymph node metastasis in CC remains unclear. In this study, we investigated the transcriptome profile of 51,507 single cells from primary tumors, positive lymph nodes (P-LN), and negative lymph nodes (N-LN) using single-cell sequencing. Validation experiments were performed using bulk transcriptomic datasets and immunohistochemical assays. Our results indicated that epithelial cells in metastatic LN were associated with cell- cycle-related signaling pathways, such as E2F targets, and mitotic spindle, and immune response-related signaling pathways, such as allograft rejection, IL2_STAT5_signaling, and inflammatory response. However, epithelial cells in primary tumors exhibited high enrichment of epithelial-mesenchymal translation (EMT), oxidative phosphorylation, and interferon alpha response. Our analysis then indicated that metastasis LN exhibited an early activated tumor microenvironment (TME) characterized by the decrease of naive T cells and an increase of cytotoxicity CD8 T cells, NK cells, FOXP3+ Treg cells compared with normal LN. By comparing the differently expressed gene of macrophages between tumor and metastatic LN, we discovered that C1QA+ MRC1^low macrophages were enriched in a tumor, whereas C1QA+ MRC1^high macrophages were enriched in metastatic LN. Finally, we demonstrated that cancer-associated fibroblasts (CAFs) in P-LN were associated with immune regulation, while CAFs in tumor underwent EMT. Our findings offered novel insights into the mechanisms of research, diagnosis, and therapy of CC metastasis.

CCR1 upregulation in macrophages promotes sequential T/B-cell infiltration via CCL5-mediated crosstalk, aggravating salivary gland damage in Sjögren's syndrome (SS), implicating CCR1/CCL5 as potential therapeutic targets. To investigate the role of CCR1/CCL5 in salivary gland lymphocytic infiltration and SS progression. Functional enrichment analysis was performed on four bulk RNA-seq datasets to identify common immune pathways, highlighting CCR1 as a key factor. Cytohubba and STRING analyses identified CCL5 as the primary ligand for CCR1 in SS salivary glands. Single-cell RNA sequencing (scRNA-seq) and temporal analyses were performed for the cellular distribution of CCR1 and CCL5, their interactions, and sequential immune cell infiltration, and further validated using multiplex immunofluorescence (mIF) in SS patients and Non Obese Diabetes (NOD) mice salivary glands. The effect of CCR1 inhibition on SS progression was assessed in NOD mice. We observed CCR1 overexpressed (CCR1^hi) macrophages and CCL5 overexpressed (CCL5^hi) macrophages and T cells in SS salivary glands. Temporal analysis revealed that CCR1^hi/CCL5^hi macrophages first infiltrated and recruited CCL5^hi T cells, followed by B cells in SS salivary glands. Interaction analysis demonstrated strong autocrine signaling in CCR1^hi/CCL5^hi macrophages and significant crosstalk between CCR1^hi/CCL5^hi macrophages, CCL5^hi T cells, and B cells. Functional enrichment indicated that CCR1^hi/CCL5^hi macrophages recruit and activate CCL5^hi T cells via chemokine signaling, and CCL5^hi T cells, in turn, recruit and activate B cells. mIF confirmed these findings in SS patients and NOD mice. CCR1 inhibition in NOD mice alleviated lymphocytic infiltration, glandular destruction, and xerostomia. CCR1^hi/CCL5^hi macrophages promote T and B cell migration in vitro. We revealed that CCR1^hi/CCL5^hi macrophages-mediated recruitment and activation of CCL5^hi T cells and B cells in salivary glands aggravate SS pathogenesis, suggesting CCR1/CCL5 as a novel target to treat SS.

Periodontitis is a common inflammatory disease that is strongly influenced by dietary habits. Coffee is one of the most common dietary components; however, current research on the relationship between coffee consumption and periodontitis, as well as its underlying mechanisms, is limited. Based on a previous report, caffeine (CA) and chlorogenic acid (CGA) were formulated into artificial coffee (AC) for this experiment. Cell viability, prostaglandin E₂ release, Western blotting, cellular reactive oxygen species (ROS) production, and NF-E2-related factor 2 (Nrf2) translocation analyses were performed to explore the effects of AC on lipopolysaccharide (LPS)-induced immortalized human oral keratinocytes (IHOKs) and elucidate their underlying mechanisms. AC pretreatment attenuated LPS-induced inflammatory mediator release, ROS production, and nuclear factor kappa B translocation in IHOKs. CA and CGA promoted AMP-activated protein kinase phosphorylation and down-regulated the nuclear factor-κB pathways to exert anti-inflammatory effects. Additionally, CGA promoted Nrf2 translocation and heme oxygenase-1 expression and showed anti-oxidative effects. Furthermore, AC, CA, and CGA components showed synergistic effects. Thus, we predict that coffee consumption may be beneficial for alleviating periodontitis. Moreover, the main coffee components CA and CGA seem to play a synergistic role in periodontitis.

Metallic molybdenum (Mo) has been increasingly recognized as a potential biodegradable metal for biomedical implants. However, the macrophage-mediated inflammatory responses to Mo-based implants remain underexplored. This study examined the in vitro inflammatory reactions of macrophages to the degradation products of biodegradable Mo implants. The short-term and long-term biodegradation behavior and the subsequent impact on cytotoxicity, metabolism, and macrophage polarization were assessed. Both Mo and its degradation products were shown to be non-toxic within macrophage tolerance limits. Nevertheless, morphological changes and pro-inflammatory polarization were observed in cells around Mo-based specimen. Notably, matrix metalloproteinase 9 (Mmp9) was identified as a key gene influencing macrophage polarization in proximity to Mo. Additionally, pre-treating the Mo specimens in culture medium for 24 h significantly mitigated its stimulatory effects on cells. These results demonstrated the significance of optimizing Mo pre-treatment methods to prevent localized inflammation associated with its degradation. Specifically, pre-treatment of Mo can effectively mitigate the adverse impacts of its early degradation on macrophages and the surrounding immune environment. Our research into these early degradation phases introduces new avenues for studying molybdenum's immunomodulatory properties, potentially through precise control of its release and the targeted expression of pivotal genes.

Osteoblasts must acquire a considerable capacity for folding unfolded and misfolded proteins (MPs) to produce large amounts of extracellular matrix proteins and maintain bone homeostasis. MP accumulation contributes to cellular apoptosis and bone disorders. Photobiomodulation therapy has been used to treat bone diseases, but the effects of decreasing MPs with photobiomodulation remain unclear. In this study, we explored the efficacy of 625 nm light-emitting diode irradiation (LEDI) to reduce MPs in tunicamycin (TM) induced-MC3T3-E1 cells. Binding immunoglobulin protein (BiP), an adenosine triphosphate (ATP)-dependent chaperone, is used to evaluate the capacity of folding MPs. The results revealed that pretreatment with 625 nm LEDI (Pre-IR) induced reactive oxygen species (ROS) production, leading to the increased chaperone BiP through the inositol-requiring enzyme 1 (IRE1)/X-box binding protein 1s (XBP-1s) pathway, and then restoration of collagen type I (COL-I) and osteopontin (OPN) expression relieving cell apoptosis. Furthermore, the translocation of BiP into the endoplasmic reticulum (ER) lumen might be followed by a high level of ATP production. Taken together, these results suggest that Pre-IR could be beneficial to prevent MP accumulation through ROS and ATP in TM-induced MC3T3-E1cells.