β-glucan is a natural polysaccharide widely existed in cereals, bacteria, and fungi. In recent years, β-glucan has attracted much attention due to its broad immunomodulatory effects. It can directly modulate neutrophils, monocytes, and macrophages, leading to their metabolic and epigenetic reprogramming and exerting a direct modulatory effect on intrinsic immunity. Besides, it can also impact the differentiation of T and B cells and the release of inflammatory factors to regulate adaptive immunity. In addition, β-glucan has been reported to influence the body's immune response by regulating gut flora. This review summarizes the immunomodulatory effects and mechanisms of β-glucan, as well as the clinical applications of β-glucan in tumors, allergic diseases, infectious diseases, and other types of diseases. This review will provide a theoretical reference and scientific basis for the further development and utilization of β-glucan.

The NOD-like receptor family pyrin domain-containing protein 3 (NLRP3) inflammasome is a macromolecular protein complex which detects danger signals and subsequently activates Caspase-1. This activation results in pyroptosis and the release of the IL-1β. Properly regulated NLRP3 inflammasome activity is essential for both sterile inflammation and host defense against microbial infections. However, aberrant activation of NLRP3 has been implicated in the pathogenesis of multiple diseases. Therefore, elucidating the mechanisms underlying NLRP3 activation is critical for advancing the understanding of how the immune system recognizes and responds to danger signals, and facilitating the development of targeted therapeutic interventions. Over the past two decades, multiple mechanistic models have been proposed, highlighting a complex and fine-tuned regulatory network. Nevertheless, the precise process of NLRP3 activation and the specific biological events it senses remain enigmatic. This review focuses on the widely accepted models of NLRP3 activation and the latest progress on its regulatory networks.

To investigate the regulation of the G protein-coupled receptor 84 (GPR84) of macrophages and its role in acute lung injury, CRISPR/Cas9 gene editing technology was used to construct Gpr84 conditional knockout mice (Gpr84^loxp/loxp), which were subsequently hybridized with macrophage-specific Lyz2-Cre mice to generate the macrophage Gpr84 conditional knockout (cKO) mice. Next, the acute lung injury model was established by a single intratracheal administration of LPS (5 mg/kg) to the conditional knockout and C57BL/6 wild-type (WT) mice. The mice were divided into 4 groups: the WT-control, the cKO-control, the WT-LPS, and the cKO-LPS groups, with 6 animals in each group. The cKO mice were identified using PCR technology. H-E staining was employed to examine the pathological changes in lung tissue. The ratio of wet weight/dry weight (W/D) was measured in lung tissue, and qRT-PCR was utilized to quantify the expression of inflammatory factors, nuclear factor erythroid 2-related factor 2 (Nrf2), and heme oxygenase 1 (Hmox1) in lung tissue. ELISA was used to determine the levels of IL-1β, IL-6, and MCP-1 in mouse serum, and immunohistochemistry was performed to detect the protein level of Nrf2 in lung tissue. The results showed that the macrophage Gpr84 knockout mice were successfully constructed, and the function of GPR84 in acute lung injury was studied. Compared to the control group, the LPS group exhibited significant inflammatory cell infiltration and alveolar collapse, increased lung injury scores, elevated lung W/D ratio, and increased expression of inflammatory factors in lung tissue and serum. However, compared to the WT-LPS group, the cKO-LPS group showed reduced pulmonary inflammatory cell infiltration, decreased lung injury scores, and significantly reduced expression of inflammatory factors in lung tissue and serum. In addition, the expressions of Nrf2 and Hmox1, which have antioxidant functions, significantly increased. This study suggests that the specific knockout of the Gpr84 gene in macrophages can significantly alleviate LPS-induced lung inflammation and enhance the resistance to oxidative stress, indicating that GPR84 may serve as a potential target for the treatment of acute lung injury.

Macrophages are innate immune cells that mainly play roles in antigen presentation and phagocytosis. Macrophages are categorized to M1 and M2 types. M2 macrophages in different microenvironments can be further divided into 4 subtypes: M2a, M2b, M2c, and M2d. M1 macrophages are induced by IFN-γ and LPS, which promote inflammation, inhibit microbial and tumor growth while M2 macrophages are activated by IL-4 or IL-13, and have anti-inflammatory activity, and their functions include tissue homeostasis maintanence, immune regulation, phagocytosis, promoting angiogenesis, and influencing tumor formation and progression. The special functions of various phenotypes of macrophages are mainly controlled by polarization signals, which can activate macrophages by upregulating different transcription programs. This review focuses on the relationships between the three metabolic pathways (sugar metabolism, lipid metabolism and amino acid metabolism) and the polarization of different macrophages which provides new solutions for diseases related to macrophage polarization.

To investigate the effect of targeted knockdown of C-C motif chemokine ligand 3 (CCL3) expression in alleviating sepsis-induced acute lung injury (ALI) and the underlying mechanism, an induced mouse sepsis model was established by cecal ligation and puncture (CLP). Lung tissue samples were collected, and differentially expressed genes and enriched pathways were identified by RNA sequencing (RNA-seq） analysis. Small interfering RNA (siRNA) technology was used to specifically knockdown the expression of the key gene CCL3, and si-CCL3 plasmids were injected via the tail vein for in vivo intervention. Mice were randomly divided into 3 groups according to the experimental design: the sham operation control group, the sepsis model group, and the si-CCL3 silencing group. H-E staining was used to evaluate the degree of lung tissue pathological damage, and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining was used to detect the level of cell apoptosis. The results showed that the sepsis mouse model was successfully established by CLP. RNA-seq results indicated that compared with the control group, there were 475 up-regulated genes and 467 down-regulated genes in the lung tissue of sepsis mice, among which the up-regulation of CCL3 was the most significant. The relative expression of CCL3 mRNA in mouse lung tissue was detected by qRT-PCR, and the results were consistent with those of the RNA-seq, showing that CCL3 was significantly up-regulated in the lung tissue of mice with sepsis. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathway analysis results showed that the NF-κB signaling pathway and ferroptosis marker proteins glutathione peroxidase 4 (GPX4) and prostaglandin-endoperoxide synthase 2 (PTGS2) were all involved in the process of sepsis-induced ALI. The results of qRT-PCR and Western blotting were consistent with RNA-seq results. After treatment with si-CCL3 plasmids, the pathological changes in the lung tissue were significantly improved, the structure of bronchi and alveoli was significantly restored, the thickness of alveolar septum was reduced, and interstitial exudation was decreased. TUNEL staining results showed that the number of apoptotic cells in the si-CCL3 silencing group was significantly lower than that of the sepsis model group (P<0.05). In conclusion, targeted knockdown of the chemokine CCL3 expression can alleviate the process of sepsis-associated acute lung injury by inhibiting the ferroptosis phenotype in lung tissue.

This study aimed to identify the B cell phenotypic characteristics in atherosclerotic plaques and explore the potential relationship between B cells and plaque inflammation progression. By comparing the immune cell profiles from single-cell sequenicing data of plaques and peripheral blood in atherosclerotic patients, 2 CD83⁺B cell clusters enriched in human atherosclerotic plaques were identified, with increased percentages of 46.74% and 19.18%, respectively. Compared to other B cell subpopulations, CD83⁺B cells expressed higher levels of aging-related genes including CDKN1A, IL-6, TNF-α, MALAT1, and GADD45B, suggesting that the increased CD83⁺B cells in plaques exhibited a senescent phenotype. To validate these findings, ApoE gene knockout (ApoE^-/-) mice were fed with Western diet (WD) to induce atherosclerosis. The proportion of CD83⁺ B cells in total B cells was significantly higher in the arterial tissues of WD mice (fold change=5.36，P=0.010 8). Consistently, in WD mice, the average fluorescence intensities of senescence-associated inflammatory cytokines including IL-6, TNF-α, and IFN-γ in CD83⁺B cells were significantly higher than those of the CD83^-B cells, with significant fold changes of 1.95 (P=0.020 2), 4.25 (P=0.003 0), and 4.20 (P=0.016 9), respectively. This study reveals that CD83⁺B cells are enriched in atherosclerotic plaques and exhibit a senescence-like phenotype, producing higher levels of senescence-associated inflammatory factors that may contribute to arterial inflammation.

In order to explore the mechanism of interaction between C-X-C motif chemokine ligand 13 (CXCL13) and glycosaminoglycan in autoimmune diseases, C57BL/6 mice were immunized with myelin oligodendrocyte glycoprotein (MOG) peptides to establish the experimental autoimmune encephalomyelitis (EAE) model. On the 3rd day post-immunization, the modeled mice were randomly divided into 3 groups: the control group, the group treated with mutant mouse CXCL13 protein which lacked the glycosaminoglycan binding capacity (the mutant group), and the group treated with mouse CXCL13 protein (the wild type group). Corresponding protein intervention was performed respectively. Disease scoring was continued. Flow cytometry was used to detect mouse spleen B cells and plasma cell subsets on day 7 and day 14 post-immunization, and the expressions of IgG and IgM in plasma and CXCL13 in spinal cord and plasma were measured by ELISA. Tissue section and transcriptome sequencing of splenic cells were analyzed. The results showed that the wild type group had delayed onset and significantly lower disease scores than the control group (P=0.031 3), while those of the mutant group was between the control group and the wild type group. On the 7th day after immunization, compared to the control group, both the mutant group (P=0.017 2) and the wild type group (P=0.000 2) showed an increased proportion of germinal center B cells and decreased proportion of plasmablasts (P=0.022 3 in the mutant group, P=0.000 1 in the wild type group); On the 14th  day after immunization, compared to the control group, both the mutant group (P=0.000 5) and the wild type group (P=0.003 1) showed an increased proportion of germinal center B cells, while the proportions of plasmablasts (P=0.005 8 in the mutant group, P=0.027 9 in the wild type group) and plasma cells decreased (P=0.002 1 in the mutant group, P=0.034 5 in the wild type group). On day 14 post-immunization, the plasma subsets basically maintained the same trend as day 7 post-immunization, but the germinal center B subsets composition changed dynamically. The plasma IgM level of the wild type group decreased significantly in the early stage of the immune response (P=0.004 6). However, on day 14 post-immunization, compared to that of the control group, the plasma IgG levels in the mutant group (P=0.006 1) and the wild type group (P=0.001 8) increased significantly. The plasma IgM levels in the mutant group (P=0.012 3) and the wild type group (P=0.009 1) also increased significantly. The plasma IgG affinity detection showed that the affinity between IgG and MOG peptides in the control group was higher than that in the other two groups (P=0.038 0). The myelin staining analysis of the spinal cord lumbar enlargement plane sections showed that the demyelination degree was consistent with the trend of disease scores. The transcriptome sequencing data of splenic cells also showed that over time, the differentially expressed genes in the mutant group were more enriched in the humoral immune pathway. This study suggests that the CXCL13-glycosaminoglycan interaction is involved in the antibody production process and mediates the immune response.

Autoimmune diseases are chronic conditions characterized by the erroneous attack of immune system towards self-tissues and organs, characterized by abnormal activation of self-reactive T and B cells, production of autoantibodies, and persistent tissue inflammation. Common examples include rheumatoid arthritis, systemic lupus erythematosus, and immune thrombocytopenia, which may lead to multi-organ dysfunction. Recent research progress has revealed significant metabolic reprogramming in immune cells associated with these diseases. Metabolic reprogramming refers to the process in which cells adjust metabolic pathways to adapt to survival needs in different microenvironments. In these diseases, metabolic reprogramming primarily manifests as a shift from oxidative phosphorylation to glycolysis. This metabolic alteration not only provides energy for cell activation but also directly influences their immunosuppressive and immunomodulatory functions. Based on these findings, researchers have developed various metabolic regulatory strategies, such as glycolysis and mTOR pathway inhibitors, offering new approaches for the diagnosis and treatment of autoimmune diseases. Advances in this field hold promise for improving patient prognosis and facilitating the development of personalized therapy.

To study the molecular mechanism by which the N6-methyladenosine (m6A)-reading protein YTHDC2 regulates macrophage M1 polarization, the expression level of YTHDC2 protein was detected in a macrophage polarization model. Next, the expressions of polarization markers was detected in YTHDC2 knockdown M1 macrophages. YTHDC2 knockdown M1 macrophages were constructed, and YTHDC2 knockdown and wild type M1 macrophages were sequenced using Illumina Novaseq JP26000/MGISEQ-T7 sequencing platform to identify the differentially expressed genes of the 2 cell groups. The differentially expressed genes were analyzed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment methods to explore the YTHDC2 regulatory pathway and screen for YTHDC2 downstream target genes. Cellular experiments were used to reveal the role of YTHDC2 downstream target gene fibulin 1 (FBLN1) in M1 macrophages. The results showed that YTHDC2 expression was elevated in M1 macrophages. Expression levels of polarization markers were decreased in YTHDC2 knockdown M1 macrophages. 147 differentially expressed genes were identified between YTHDC2 knockdown and wild type M1 macrophages. FBLN1 expression is up-regulated in YTHDC2 knockdown M1 macrophages. Potential m6A methylation might occur at multiple sites in FBLN1. Expression levels of polarization markers were increased in M1 macrophages with the knockdown of YTHDC2 and FBLN1. In conclusion, YTHDC2 may promote macrophage M1 polarization by regulating the m6A modification of FBLN1.

Macrophage extracellular traps (MET) are reticular structures composed of DNA and protein components secreted by activated macrophages. The formation process mainly includes mechanisms such as nicotinamide adenine dinucleotide phosphate oxidase (NOX)/reactive oxygen species (ROS)-dependent or -independent systems and histone citrullination mediated by peptidyl arginine deiminases (PAD). It is a new immune mechanism of macrophages, which can capture and kill bacteria, fungi, parasites, and other pathogenic organisms. However, some pathogenic organisms can escape the effect of MET during infection. This review mainly discusses the formation mechanism of MET and their role in resistance to pathogens, aiming to provide new targets for the treatment of diseases caused by pathogens.

IL-17D is a member of the IL-17 cytokine family which consists of 6 members, including IL-17A to IL-17F. The IL-17 receptor （IL-17R） family consists of 5 members: IL-17RA to IL17RE. IL-17A, as the typical representative of the IL-17 family and a hallmark cytokine produced by Th17 cells, plays a significant role in autoimmune diseases, tumorogenesis and inflammatory response. Although IL-17D is similar to IL-17A in the potential to elicit inflammatory cytokine production, studies on IL-17D are relatively fewer compared to those of IL-17A, and its function and mechanism of action remain elusive. In recent years, it has been found that IL-17D plays an important role in various diseases such as inflammatory response, viral infection, tumor and cardiovascular diseases, etc. By interacting with receptors on the surface of target cells, IL-17D activates downstream signaling pathways, regulates the expression of inflammatory factors, chemokines, etc., and participates in the recruitment and activation of immune cells. In addition, IL-17D has potential therapeutic value in antiviral immunity and tumor immunosurveillance. The review summarizes the structural features, biological activity and expression of IL-17D and its correlation with diseases.

The aim of this research is to explore the effect of adenosine A2A receptor (A2AR) blockade on the angiogenesis in thyroid cancer by regulating macrophage polarization. THP-1 cells were induced to differentiate into macrophages by phorbol ester which were subsequently divided into the shA2AR group and the shA2AR-NC group. The shA2AR and shA2AR-NC lentivirus was transfected into the two groups respectively. Western blotting was used to verify the blocking efficiency of A2AR, and flow cytometry was used to detect the polarization status of macrophages in each group. Next, the two groups of macrophage were co-cultured with human thyroid cancer cell TPC-1 for 24 h. The supernatants of TPC-1 co-culture were added into the corresponding human umbilical vein endothelial cells (HUVECs) groups: the shA2AR co-culture supernatant group and the shA2AR-NC co-culture supernatant group. The angiogenesis was measured and the expressions of vascular endothelial growth factor A (VEGFA) , vascular endothelial cadherin (VE-cadherin) and the activation of Notch1/JAK2/STAT3 signaling pathway were detected by Western blotting. The proliferation of HUVECs was detected by CCK-8 assay. The results showed that the expression of A2AR in macrophages was blocked by lentivirus transfection, and the macrophages were mainly polarized to M1 type. Compared to that of the shA2AR-NC co-culture supernatant group, the HUVECs proliferation activity in the shA2AR co-culture supernatant group was significantly decreased (P<0.01), as well as the expressions of VEGFA and VE-cadherin (P<0.01). The number of blood vessel formed was significantly reduced (P<0.01), and the activation of the Notch1/JAK2/STAT3 signaling pathway was significantly inhibited. Upon Notch1 receptor agonist Jagged1 treatment, the HUVECs in the shA2AR co-culture supernatant group showed increased activation of the Notch1/JAK2/STAT3 signaling pathway (P<0.01). In summary, the study reveals that blocking A2AR may induce macrophage M1 type polarization. By inhibiting the activation of the Notch1/JAK2/STAT3 signaling pathway, A2AR blockade reduces HUVECs proliferation and angiogenesis of thyroid cancer.

This study explores the potential mechanism by which dihydroartemisinin (DHA) modulates the complement C3/C3a receptor (C3aR) signaling pathway to improve neuroinflammation in the hippocampus in a mouse depression model. Forty male C57BL/6J mice aged 8 weeks were randomly divided into 4 groups: the control group, the model group, the low-dose DHA (DHA-L) group, and the high-dose DHA (DHA-H) group, with 10 mice in each group. Except for the control group, the remaining groups were subjected to chronic unpredictable mild stress (CUMS) to establish the depression model. Behavioral assays were performed to evaluate the success or failure of the model and changes in depressive behaviors in the mice. H-E staining and Nissl staining were used to assess neuronal pathological damage in the hippocampus. ELISA was employed to detect inflammatory cytokine levels, while immunohistochemistry was used to observe microglial activation. Western blotting was performed to detect the expression levels of C3 and C3aR proteins. The results showed that the depression model in mice was successfully established. Mice in the model group showed a significant decrease in sucrose preference, time spent in the center of the open field, and total distance traveled in the open field compared to the control group (P<0.01). In the tail suspension and forced swimming tests, the immobility time of modeled mice was significantly prolonged (P<0.001). Mice in the DHA-L group and the DHA-H group showed a significant increase in sucrose preference, time spent in the center of the open field, and total distance traveled compared to the model group (P<0.05). Furthermore, their immobility time in the tail suspension and forced swimming tests was significantly reduced (P<0.05). The inflammatory cytokine levels in the hippocampus of the model mice were significantly higher than those in the control group (P<0.01), and DHA treatment significantly reversed this effect (P<0.05). DHA treatment also alleviated the pathological damage to hippocampal neurons in the model mice. The number of activated microglia in the hippocampus of the model group was significantly increased compared to that of the control group (P<0.001), whereas DHA treatment significantly reduced microglial activation compared to the model group (P<0.05). The expression levels of C3 and C3aR in the hippocampus of model mice were significantly higher than those in the control group (P<0.01), and DHA treatment significantly downregulated the expression of these proteins (P<0.05). In conclusion, the antidepressant effect of DHA may be mediated by reducing neuroinflammation in the hippocampus, and the mechanism is likely associated with the C3/C3aR signaling pathway.

The aim of the study is to investigate the mechanism by which C-C motif chemokine ligand 2 (CCL2) regulates the progression of cervical cancer (CC) through ferroptosis in the tumor microenvironment (TME). Cervical cancer-derived HeLa cells were transfected with shRNA targeting CCL2 (sh-CCL2#1) and CCL2 over-expressing cDNA (ova-CCL2) lentivirus, respectively. The in vitro proliferation of tumor cells was evaluated by the CCK-8 assay, and the metastasis and invasion abilities were assessed by the Transwell assay. Phorbol 12-myristate 13-acetate (PMA) was used to induce THP-1 differentiation, which was co-cultured in a 1∶4 ratio for 48 h with HeLa cells transfected with lentivirus mentioned above. Western blotting and qRT-PCR were used to measure the expression of macrophage polarization markers. The co-cultured cells in each group were further treated with ferroptosis inducer erastin, and the expressions of ferroptosis markers in macrophages were detected. The results showed that compared with the NC group, the HeLa cells of CCL2 over-expression group showed macrophage-dependent tumor-promoting phenotype (P<0.001). The macrophages in the TME underwent M2-type polarization, which was reversed upon the administration of erastin. In summary, overexpression of CCL2 in HeLa cells promotes the M2-type polarization of macrophages in the TME, which in turn stimulates the proliferation, migration, and invasion of HeLa cells. Reversion of the macrophage polarization by ferroptosis inducers disrupts the TME and inhibits tumor cell aggressive behavior. Therefore, inducing macrophage ferroptosis in the TME may help to suppress tumor progression.

IgA vasculitis (IgAV) is a common vasculitis syndrome during childhood and is considered a self-limiting disease. While its pathogenesis remains largely elusive. The short-term symptoms primarily involve skin, gastrointestinal tract and joint burden, with a generally favorable prognosis. However, in the long course, severe renal damage can progress to renal failure. Therefore, investigating the pathogenesis of IgAV is crucial for preventing the disease and managing related organ damage. Accumulating evidences indicate a strong correlation between galactose-deficient IgA1 (GD-IgA1) and the occurrence of IgAV. This review comprehensively discusses the relationship between GD-IgA1 and pediatric IgAV and  firstly  elucidates the central role of GD-IgA1 in IgAV pathogenesis from multiple dimensions, including genetics, immune cells, infection, and the gut microbiota. This review also elaborates the mechanisms underlying the formation and deposition of immune antibody complexes and their sequential damage to kidney and gastrointestinal tract which provide a theoretical foundation for deeper understanding of the disease pathogenesis and the development of glycosylation targeted and microbiome-based interventions.

This study aims to explore the effect of Qianjin Weijing Decoction on acute lung injury in sepsis rats through pyroptosis mediated by NF-κB/NOD-like receptor family pyrin domain-containing protein 3(NLRP3) signaling pathway. Rat model of sepsis was established by cecal ligation. Rats were randomly divided into the model group, the low-dose, medium-dose and high-dose Qianjin Weijing Decoction groups, and the Qianjin Weijian Decoction + phorbol 12-myristate 13-acetate (PMA; NF-κB signaling pathway activator) group, with 12 rats in each group. An other 12 rats from the same cohort without cecal ligation were assigned as the sham operation group. The lung index and water content of lung tissue were measured. The pathological changes of lung tissue were observed by H-E staining, and the relevant scoring was calculated. The pyroptosis of lung tissue was detected by immunofluorescence staining. The levels of inflammatory factors in serum were detected by ELISA. The expressions of NF-κB/NLRP3 signaling pathway related proteins were detected by Western blotting. The results showed that compared to those of the sham operation group, the lung index, water content of lung tissue, score of lung injury, relative fluorescence intensity of NLRP3, levels of serum IL-1β, IL-18, and TNF-α, expressions of phosphorylated NF-κB p65 (p-NF-κB p65)/NF-κB p65, NLRP3 and cleaved Caspase -1 (C-Caspase-1) protein in lung tissue were all increased in the model group (P<0.05). Compared to those of the model group, the lung index, water content of lung tissue, score of lung injury, relative fluorescence intensity of NLRP3, levels of serum IL-1β, IL-18, and TNF-α, expressions of p-NF-κB p65/NF-κB p65, NLRP3 and C-Caspase-1 in lung tissue were all decreased in the medium-dose and high-dose Qianjin Weijing Decoction groups (P<0.05). The treatment of PMA reversed the above effects of Qianjin Weijing Decoction. This study shows that Qianjin Weijing Decoction may inhibit pyroptosis by down-regulating the NF-κB/NLRP3 signaling pathway, thus alleviate acute lung injury in sepsis rats.

This study aims to elucidate the inhibitory effects and underlying mechanisms of allicin on fibrosis in mouse cardiac fibroblasts (CF) through long non-coding RNA (lncRNA) maternally expressed gene 3 (MEG3). Mouse CFs were isolated and cultured, and an in vitro fibrosis model of CFs was established. The fibrotic CFs were treated with allicin at concentrations of 10, 20, and 40 μg/L to evaluate the protective effect of allicin on CF injury. Subsequently, cells were transfected with the sh-lncRNA MEG3 plasmid or the sh-lncRNA NC plasmid. The cells were divided into the control group, the model group, the allicin group, the sh-lncRNA MEG3 group, the sh-lncRNA NC group, the allicin + sh-lncRNA MEG3 group, and the allicin + sh-lncRNA NC group. CCK-8 assay was used to evaluate cell proliferation, flow cytometry was employed to assess cell apoptosis, and qRT-PCR was conducted to measure the expression changes of matrix metalloproteinase (MMP)-2, MMP-3, MMP-9, lncRNA MEG3, and inflammatory cytokines (IL-8, IL-6, IL-1β). Western blotting was used to detect the expressions of alpha-smooth muscle actin (α-SMA), collagen type Ⅰ (COL1), collagen type Ⅲ (COL3), and proteins involved in the NF-κB signaling pathway (NF-κB p65, p-NF-κB p65, inhibitor of κB ［IκB］ , and p-IκB). The results showed that compared to the control group, the model group showed enhanced cell proliferation (P<0.01), reduced apoptosis rate (P<0.01), upregulated lncRNA MEG3 levels (P<0.01), and increased mRNA levels of MMP-2, MMP-3, MMP-9, as well as IL-8, IL-6, and IL-1β (P<0.01). After allicin treatment, cell proliferation was inhibited (P<0.01), apoptosis rate increased (P<0.01), and the mRNA levels of MMP-2, MMP-3, MMP-9, as well as IL-8, IL-6, and IL-1β were downregulated (P<0.05) in a dose-dependent manner. In addition, allicin downregulated the protein expressions of α-SMA, COL1, and COL3, as well as the p-NF-κB p65/NF-κB p65 ratio (P<0.01), while upregulating the p-IκB/IκB ratio (P<0.01) in the CFs fibrosis model. Compared to the model group, both the sh-lncRNA MEG3 group and the allicin group showed similar trends. Moreover, the combined treatment of allicin and sh-lncRNA MEG3 generated stronger inhibitory effect on CF injury than of sh-lncRNA MEG3 alone. This study suggests that allicin exerts a protective effect against fibrosis induced CF injury by downregulating lncRNA MEG3, thereby inhibiting the NF-κB pathway to suppress CFs proliferation, and reduce the levels of MMPs, inflammatory cytokines, and collagen deposition proteins.

This study aimed to investigate the role of tripterygium glycosides (TG) in alleviating the pathological damage and inflammation in the colonic tissue of ulcerative colitis (UC) mice. A total of 60 BALB/C mice were randomly divided into 4 groups: the control group, the model group, the low-dose TG group, and the high-dose TG group, with 15 mice in each group. All groups, except the control group, were treated with 3% dextran sulfate sodium (DSS) solution for 7 consecutive days to induce the UC model. The treatment groups received TG by oral gavaging for 7 days, during which body weight, fecal condition, and disease activity index (DAI) were monitored. After the treatment, colonic tissue pathology was assessed using H-E and periodic acid-Schiff (PAS) stainings, and levels of serum inflammatory cytokines were measured by ELISA. Western blotting was used to detect the protein expressions of tight junction protein 1 (ZO-1), occludin, NOD-like receptor family pyrin domain-containing protein 3 (NLRP3), apoptosis-associated speck-like protein (ASC), Caspase-1, IL-1β, and IL-18 in the colon tissue. The results showed that TG treatment significantly reduced the DAI score (P<0.05), promoted the recovery of colonic length (P<0.01), and alleviated inflammatory damage in colonic tissue. Serum levels of TNF-α, IL-1β, and IL-6 were significantly reduced, while the level of IL-10 elevated (P<0.05). Furthermore, TG treatment significantly decreased the protein expressions of NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in colonic tissue (P<0.05) while increased the expressions of ZO-1 and occludin (P<0.05). In conclusion, TG may alleviate symptoms of UC mice by modulating the NLRP3/Caspase-1 signaling pathway to inhibit inflammation and restore intestinal barrier function.

The study aims to explore the role of E2F transcription factor 8 (E2F8) on glycolysis and tumor immunosuppression in breast cancer cells. Tissues from 12 breast cancer patients were collected and E2F8 expression was detected by immunohistochemistry and Western blotting. The E2F8 expressions of human breast cancer cell lines and the normal human breast epithelial cell line were measured by Western blotting. Stable E2F8- and serum amyloid A1 (SAA1)-overexpressing MCF-7 cell lines and negative control cell lines were constructed, as well as stable E2F8- and SAA1-knockdown T47D cell lines and negative control cell lines. Western blotting was used to detect the expressions of glycolysis-related proteins including lactate dehydrogenase A (LDHA), hexokinase 2 (HK2), and pyruvate kinase M2 (PKM2). qRT-PCR was employed to measure the protein levels of IL-6, TNF-α ，and C-X-C motif chemokine ligand 10 (CXCL10) mRNA in cells. THP-1 cells were differentiated into M0 macrophages by phorbol ester induction. M0 macrophages were co-cultured with E2F8-knockdown T47D cells and E2F8-overexpressing MCF-7 cells, and the M1/M2 polarization was analyzed by flow cytometry. The results showed that E2F8 expression was upregulated in tumor tissues as well as all tumor cell lines. In E2F8- and SAA1-overexpressing cells, the protein levels of SAA1, LDHA, HK2, and PKM2 were significantly increased, while the knockdown cells showed the opposite results. When co-cultures with E2F8-overexpressing cells, the proportion of M1 macrophages was significantly reduced (P<0.001), while M2 ratio was significantly increased (P<0.01). In contrast, the knockdown group showed the opposite results (P<0.05). In the E2F8-overexpressing group, IL-6, TNF-α, and CXCL10 mRNA levels were significantly decreased (P<0.01), while in the knockdown group, the opposite results were observed (P<0.01). This study suggests that E2F8 enhances glycolysis in breast cancer cells by promoting SAA1 protein expression and release.

As an important part of the innate immune system, immune responses of macrophages are triggered by PAMP recognized by PRR, which have important functions in host defense and maintenance of tissue homeostasis. Bacterial infection can induce macrophage death through a variety of pathways, including apoptosis, pyroptosis, necroptosis, and ferroptosis, which affect the host immune response and disease process. Different types of cell death have different effects on host immune response and bacterial clearance. Bacteria can evade immune clearance by regulating macrophage death, leading to chronic or systemic infections. This review focuses on different mechanisms by which bacterial infection regulates macrophage death and its impact on the host immune response. Future studies on the molecular mechanisms of macrophage death under different bacterial infection conditions and their regulation of the host immune response may help to develop new therapeutic strategies, such as targeted drugs or cell therapies, to deal with the challenges of drug-resistant bacteria and systemic infections.

The aim of the study was to investigate the effect of human umbilical cord blood mesenchymal stem cell (hUC-MSC) on immune thrombocytopenia (ITP) mouse model and its regulation on the retinoic acid-related orphan receptor γt (ROR-γt)/forkhead winged helix transcription factor 3 (Foxp3) signaling pathway. Healthy hUC-MSCs were isolated and cultured, the growth patterns were observed and identified, and the stably-proliferating primary cells were selected for passaging. Thirty male BALB/c mice were randomly divided into the normal control group (Con group), the ITP model group and the hUC-MSC group, with 10 mice in each group. After corresponding treatments, the general condition of the mice was observed. Peripheral platelet count (PLT) was measured, and serum levels of IL-6, IL-17, IL-10, and TGF-β1 were detected using ELISA. Wright-Giemsa staining was used to evaluate the pathological changes in mouse bone marrow and to count the megakaryocytes. Western blotting and RT-qPCR were used to detect protein and mRNA levels of ROR-γt and Foxp3 in mice spleen and peripheral blood, respectively. The results showed that compared to those of the Con group, the serum IL-6 and IL-17 levels, ROR-γt mRNA and protein expression in spleen and the ROR-γt mRNA and protein expression in peripheral blood were all significantly higher in the ITP model group (all with P＜0.05), while the PLT, serum IL-10 and TGF-β1 levels, Foxp3 mRNA and protein expression in spleen and Foxp3 mRNA and protein expression in peripheral blood were significantly decreased (all with P＜0.05), along with significantly reduced number of platelet-producing megakaryocytes (P＜0.05). Compared to the ITP model group, the hUC-MSC group showed significantly lower serum IL-6 and IL-17 levels, splenic ROR-γt mRNA and protein expression and peripheral blood ROR-γt mRNA and protein levels (all with P＜0.05), but significantly higher PLT, serum IL-10 and TGF-β1 levels, splenic Foxp3 mRNA and protein expression, peripheral blood Foxp3 mRNA and protein expression (all with P＜0.05), and the elevated number of platelet-producing megakaryocytes (P＜0.05). As a result, hUC-MSC can promote platelet formation and increase the number of platelet-producing megakaryocyte, which may relief the symptoms of ITP mice by regulating the expressions of cytokines and the ROR-γt/Foxp3 axis.

A preliminary quantitative detection method for heparin-binding protein (HBP) based on fluorescence immuno-chromatography was established. A sandwich immunoassay format was utilized to optimize the antibody concentrations (coated antibody: 0.5-2.0 mg/mL，labeled antibody: 4-12 μg/mL) and reaction time (5-22 min) to establish the HBP fluorescence immunochromatographic assay. Upon successful establishment of the assay, performance evaluations were conducted including precision, accuracy, interference testing, stability, and method comparison. The results demonstrated a linear range of 4-500 ng/mL, with stable signals observed after 18 min of chromatography. Intra-assay and inter-assay coefficients of variation (CV) were ≤10% and ≤15%, respectively, while accuracy deviations were restricted within ±10%. Interference experiments confirmed that hemoglobin, lipids, bilirubin, citrate, EDTA, procalcitonin (PCT), and CRP exhibited no significant interference (P>0.05), whereas heparin significantly affected the results (P<0.001). Accelerated stability testing at 37 ℃ revealed no significant signal changes over an 8-week storage period (P>0.05). Comparative analysis with a commercially available HBP analyzer (Joinstar) confirmed a strong correlation (r=0.997 3) and no significant difference (P>0.05) between the two methods. This newly developed fluorescence immunochromatographic method features user-friendly operation, robust anti-interference capability and meets clinical requirements for point-of-care quantitative HBP detection, which will provide an auxiliary diagnostic tool for infectious diseases. 

The aim of the study is to investigate the effects of icariin on apoptosis of retinal cells in diabetic retinopathy (DR) rats through the silent mating-type information regulation 2 homolog 1 (SIRT1)/forkhead-box transcription factor O3a (FOXO3a)/NF-κB signaling pathway. The SD rats were randomly divided into the control (Ctrl), the DR, the low-dose icariin (L-icariin), the high-dose icariin (H-icariin), and the SIRT1 inhibitor + H-icariin groups. The DR rat model was established by intraperitoneal injection of streptozotocin (STZ) combined with high-sugar+high-fat diet. Serum levels of oxidative stress markers (superoxide dismutase ［SOD］ and malondialdehyde ［MDA］), and inflammatory factors (TNF-α and IL-6) were measured. Retinal histopathological changes were observed by H-E staining. Retinal cell apoptosis was assessed by TUNEL assay. B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) expressions were detected by immunohistochemistry. Western blotting was used to analyze the protein expressions of SIRT1 and acetylated NF-κB (ac-NF-κB). Immunoprecipitation (IP) was used to detect ac-FOXO3a protein expression in retinal tissue. The results showed that compared with the Ctrl group, the DR group exhibited reduced serum SOD level and increased expressions of MDA, TNF-α, and IL-6 (all with P＜0.05). Retinal cells displayed disordered arrangement and were significantly decreased in number, accompanied by cellular edema and nuclear shrinkage. The apoptosis rate of retinal cells was increased (P＜0.05), along with decreased Bcl-2 and increased Bax protein expressions in retinal tissue (all with P＜0.05). After intervention with L-icariin or H-icariin, these abnormalities were mitigated, with the H-icariin group showing better efficacy. Compared with the Ctrl group, the DR group demonstrated reduced SIRT1 protein expression and increased ac-FOXO3a and ac-NF-κB protein expressions (all with P＜0.05) in retinal tissue. Both H-icariin intervention and concurrent SIRT1 inhibitor treatment diminished or reversed these protein expression profiles. Notably, SIRT1 inhibitor reduced the effect of icariin on retinal cell apoptosis. These findings suggest that icariin may alleviate retinal lesions and inhibit retinal cell apoptosis in DR rats through regulating the SIRT1/FOXO3a/NF-κB signaling pathway.

The study aims to investigate the regulation of piperine (PIP) on insulin resistance in type 2 diabetes mellitus (T2DM) rats by the insulin receptor substrate 1 (IRS-1)/Akt/PI3K signaling pathway. Wistar rats were randomly assigned to the control group, the T2DM group, the low dose PIP (PIP-L) group (3 mg/kg), the high dose PIP (PIP-H) group (30 mg/kg), the IRS-1 inhibitor NT157 group (2 μmol/L NT157), the PIP-L+NT157 group (3 mg/kg PIP+2 μmol/L NT157), and the PIP-H+ NT157 group (30 mg/kg PIP+2 μmol/L NT157). Except for the control group, rats in all the other groups were fed with high-fat and high fructose and injected with streptozotocin to establish the T2DM model. Fasting blood glucose, serum insulin, triglycerides, total cholesterol, high-density lipoprotein cholesterol, aspartate transaminase, and alanine transaminase levels were measured. H-E staining was used to detect the pathological changes in pancreatic tissue. Western blotting was used to detect the expressions of inflammatory factors TNF-α, IL-6, CRP, and IRS-1/Akt/PI3K pathway related proteins in pancreatic tissue. The results showed that the pancreatic tissue in the NT157 group and T2DM group exhibited obvious damage. And the fasting blood glucose, insulin resistance, triglyceride, total cholesterol, aspartate transaminase, alanine transaminase, and inflammatory factors were higher than those of the control group (P<0.05)，while the serum insulin, high-density lipoprotein cholesterol, p-IRS-1/IRS-1 and p-Akt/Akt ratios, and PI3K protein expression were all lower (P<0.05). In contrast, the PIP-L and PIP-H groups showed significant improvement of the above indicators（P<0.05）. The IRS-1 inhibitor NT157 partially reversed the beneficial effects of both low or high PIP treatments. This study suggests that PIP may ameliorate insulin resistance in T2DM rats by activating the IRS-1/Akt/PI3K signaling pathway. 

The study aims to analyze the expressions of non-structural maintenance of chromosome condensin Ⅰ complex subunit D2 (NCAPD2) and a disintegrin and metalloprotease 17 (ADAM17) in colon cancer tissues and the correlation with patient prognosis. One hundred colon cancer patients subjected to radical resection surgery were selected. During the surgery, colon cancer tissue specimens and normal tumor-adjacent tissue specimens with a distance of more than 5 cm from the colon cancer tissue were collected from the patients. The expression levels of NCAPD2 and ADAM17 in the two types of specimens were detected by Western blotting, and the expressions of NCAPD2 and ADAM17 in patients with different clinical and pathological characteristics were analyzed. The patients were followed up for 3 years and their survival data were recorded. The pateints were divided into the survival group (67 cases) and the death group (33 cases). The expression levels of NCAPD2 and ADAM17 were compared between the two groups. ROC curve was used to evaluate the predictive value of NCAPD2 and ADAM17 for the prognosis of colon cancer patients. The cut-off value in the curve was used as the threshhold for NCAPD2 and ADAM17 expressions, which was used to divide patients into high expression and low expression groups. Meanwhile, Kaplan-Meier survival curve was used to compare the survival of the two groups. The results showed that the expression levels of NCAPD2 and ADAM17 in colon cancer tissues were higher than those in normal tissues (P<0.001). The expression levels of NCAPD2 and ADAM17 in patients with infiltration degree T3-T4, low differentiation, distal metastasis, and lymph node metastasis were higher than those in the corresponding control group (P<0.05). The expression levels of NCAPD2 and ADAM17 in the death group were higher than those in the survival group (P<0.05). The ROC curve showed that the AUC of NCAPD2 and ADAM17 for predicting death were 0.780 and 0.721, respectively. The Kaplan-Meier survival curve showed that the survival time of patients with high expressions of NCAPD2 and ADAM17 was significantly shorter than that of patients with low NCAPD2 and ADAM17 expressions (P<0.001). The study suggests that NCAPD2 and ADAM17 are highly expressed in colon cancer tissues, and are highly correlated to the invasion, metastasis and prognosis of colon cancer, underscoring their significance in the prognosis assessment of colon cancer patients.

The study aimed to explore the regulation of N6-methyladenosine (m6A) modification of SOCS2 on the progression of colorectal cancer via the JAK2/STAT3 signaling pathway and the possible underlying mechanism. Human normal colon epithelial cell NCM460 and human colon cancer cells SW480, LOVO, HCT-15, HT-29, and SW1116 were cultured, and the overall level of m6A RNA was detected. Western blotting and RT-qPCR were used to detect METTL3 protein and mRNA levels, respectively, and SELECT-qPCR modified quantitative technique was used to detect the m6A level at SOCS2 site. LOVO and HT-29 cells were divided into the control group (WT cells) and the METTL3 low expression group (transfection with low-expression METTL3 plasmid). SOCS2 levels in LOVO and HT-29 cells were detected by immunofluorescence staining. Western blotting was used to detect the expression levels of JAK2, STAT3, p-JAK2 and p-STAT3 in LOVO and HT-29 cells. Plate clone formation assay was used to detect the clone-forming ability of LOVO and HT-29 cells, and scratch assay was used to detect the migration of LOVO and HT-29 cells. The invasive ability of LOVO and HT-29 cells was detected by the Transwell assay. The results showed that, compared to those of NCM460, the overall levels of m6A RNA in all the cancer cell lines were increased (P<0.05, P<0.01), and the protein and mRNA levels of METTL3 in LOVO and HT-29 cells were also increased (P<0.01). The m6A level of SOCS2 was increased (P<0.05). Compared to that of the control group, the expression of SOCS2 in LOVO and HT-29 cells in the METTL3 low expression groups were increased (P<0.01), while the expression of METTL3 protein and mRNA were decreased (P<0.01), and no significant changes in JAK2 and STAT3 (P>0.05) were observed. The expressions of p-JAK2 and p-STAT3 were decreased (P<0.01), and the proliferation, migration, and invasion abilities of LOVO and HT-29 cells were also decreased (P<0.01). This study indicates that low expression of METTL3 can downregulate JAK2/STAT3 signaling pathway and inhibit the proliferation, migration, and invasion of colon cancer cells, which may be related to the regulation of JAK2/STAT3 signaling pathway by m6A modification of SOCS2.

This study aimed to investigate the role of tropomyosin 1 (TPM1) and matrix metalloproteinase 9 (MMP-9) on the malignant behavior of bladder cancer cells through the NF-κB/MMP-9 signaling pathway. The Cancer Genome Atlas (TCGA) and the TIMER database were used to analyze the expression differences of TPM1 in bladder cancer tissues and normal bladder tissues. Survival analysis tools were employed to assess the association between TPM1 and the prognosis of bladder cancer patients. Additionally, the correlation between TPM1 and immune cell infiltration across various cancers was evaluated using the TIMER database and R packages.  T-24 and THP-1 cells were used in in vitro experiments. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of THP-1 cells to macrophage.  Overexpression or silencing vector of TPM1, combined with NF-κB/MMP-9 pathway agonist/antagonist interventions and T-24 and THP-1 cell co-culture system were conducted. Western blotting was used to detect the expression of TPM1, NF-κB, MMP-9, and macrophage polarization markers (IL-6, iNOS, Arg-1, and CD206). MTT assay, scratch wound healing assay, and Transwell invasion assay were performed to evaluate the proliferation, migration, and invasion of T-24 cells. Western blotting was also employed to examine the expression of epithelial-mesenchymal transition (EMT) related proteins (E-cadherin, N-cadherin, and Vimentin). The results showed that the positive expression rate of TPM1 protein in bladder cancer tissues was lower than that in normal bladder tissues (P<0.01). Patients with low TPM1 expression had significantly worse survival rates compared to those with high expression (P<0.01). TPM1 expression was negatively correlated with M2 macrophage infiltration (P<0.01) but positively correlated with M1 macrophage infiltration in bladder cancer tissues. Compared with adjacent normal tissues, TPM1 protein expression was significantly reduced in bladder cancer tissues, while its mRNA levels showed a significant decreasing trend along with the pro-inflammatory factors IL-6 and iNOS (all P<0.05). Meanwhile, the mRNA expression of M2 macrophage marker genes such as Arg-1, and CD206 was significantly upregulated (all P<0.05). Overexpression or silencing of TPM1 in T-24 cells revealed that TPM1 overexpression promoted M1 markers (IL-6, iNOS; both P<0.05) while suppressing M2 markers (Arg-1, CD206; both P<0.05). Conversely, TPM1 silencing reversed these effects. Mechanistically, TPM1 overexpression inhibited NF-κB/MMP-9 pathway activation (reduced protein levels, both P<0.05), whereas silencing enhanced pathway activity, effects reversible by NF-κB agonism/antagonism. Functional assays demonstrated that TPM1 overexpression attenuated T-24 cell proliferation, migration, invasion, and EMT (E-cadherin increased, N-cadherin/Vimentin decreased; all with P<0.05), while TPM1 silencing exacerbated malignant phenotypes. These findings indicate that TPM1 suppresses bladder cancer progression by inhibiting NF-κB/MMP-9-mediated M1-type macrophage polarization and EMT, highlighting its potential as a therapeutic target.

This study aims to investigate whether homeobox containing 1 (Hmbox1) promotes cardiomyocyte pyroptosis and aggravates myocardial ischemia-reperfusion injury (MIRI) through the PI3K/Akt signaling pathway. 24 SD rats were randomly divided into the sham operation group, the model group, the sh-NC group (lentivirus blank vector), and the sh-Hmbox1 group (lentivirus vector containing sh-Hmbox1), with 6 rats in each group. Mice of the sh-NC group and sh-Hmbox1 group were injected with corresponding lentiviral vectors 48 h before modeling. Except for the sham-operation group, the MIRI model was established by ligating the left anterior descending branch of the coronary artery through thoracotomy in the other three groups. Left ventricular function indexes ( left ventricular end-diastolic diameter ［LVEDD］, left ventricular end-systolic diameter ［LVESD］, left ventricular ejection fraction ［LVEF］, and left ventricular fractional shortening ［LVFS］) were measured by echocardiography 24 h after modeling. The levels of serum myocardial injury markers (creatine kinase-MB ［CK-MB］, cardiac troponin I ［cTnI］, lactate dehydrogenase ［LDH］) and inflammatory factors (IL-18, IL-1β) were detected by ELISA. Immunohistochemistry （IHC） was used to detect the level of Hmbox1 positive cells. Western blotting was used to detect the expressions of p-PI3K/PI3K, p-Akt/Akt, NOD-like receptor family pyrin domain-containing protein 3 (NLRP3), cleaved-Caspase-1, and GSDMD-N. The levels of oxidative stress (reactive oxygen species ［ROS］ and malondialdehyde ［MDA］) were detected by commercial kits. The results showed that, compared to the sham-operation group, the model group showed evident myocardial inflammatory cell infiltration and loose arrangement of myocytes. The levels of LVEDD, LVESD, serum CK-MB, cTnI, LDH, IL-18, and IL-1β were significantly higher in the model group than those in the sham operation group. The levels of ROS, MDA, Hmbox1 positive cells, and protein expressions of NLRP3, cleaved-Caspase-1, and GSDMD-N  in cardiac tissue were increased, while the protein expressions of p-PI3K/PI3K and p-Akt/Akt  were decreased. The values of LVEF and LVFS were reduced (all P<0.05). Compared to those of the sh-NC group, the morphology of cardiomyocytes in the sh-Hmbox1 group was improved, and the infiltration of inflammatory cells was significantly reduced. The sh-Hmbox1 group also showed decreased values of LVEDD and LVESD, along with levels of CK-MB, cTnI, LDH, IL-18, and IL-1β in serum, levels of ROS, MDA, Hmbox1 positive cells, and protein expressions of NLRP3, cleaved-Caspase-1, and GSDMD-N in cardiac tissue. On the other hand, the values of LVEF and LVFS, as well as the protein expressions of p-PI3K/PI3K and p-Akt/Akt, were significantly increased (all P<0.05). The results of this study show that knockdown of Hmbox1 inhibits cardiomyocyte pyroptosis by promoting the PI3K/Akt signaling pathway, resulting in alleviation of MIRI.  This may provide a new target for clinical treatment and prevention of MIRI.

Platelet transfusion refractoriness (PTR) is a common challenge in clinical transfusion medicine, with immune-mediated PTR receiving significant attention due to its complex pathogenesis and difficulties in clinical management. In recent years, in-depth research on the roles of HLA and human platelet antigen (HPA) in PTR, along with the rapid development of novel detection technologies, has led to significant progress in the diagnosis and treatment of immune-mediated PTR. However, accurately identifying high-risk patients, optimizing individualized transfusion strategies, and exploring effective immunomodulatory therapies remain critical issues to be addressed. This review systematically summarizes the pathophysiological mechanisms, the latest diagnostic technologies, and treatment strategies for immune-mediated PTR, aiming to provide a scientific basis for clinical practice as well as outlining future research directions to improve clinical outcomes for PTR patients.

The study aims to investigate the effects of cryptotanshinone (CTS) on neuroinflammation in schizophrenia (SP) rats. SD rats were randomly divided into the control group, the model group, the low-dose CTS group (15 mg/kg), the medium-dose CTS group (30 mg/kg), the high-dose CTS group (60 mg/kg), and the activator group (60 mg/kg CTS+10 mg/kg NOD-like receptor family pyrin domain-containing protein 3 ［NLRP3］/Caspase-1 signaling pathway activator BMS-986299), with 12 animals per group. The SP rat model was constructed by intraperitoneal injection of dizocilpine in all groups except the control group. H-E staining was used to observe the pathological damage of hippocampal tissue. ELISA was used to detect the expression levels of serum IL-6 and TNF-α. The activities of hippocampal tissue acetyl choline (dihydrokaempferol, Ach) and acetyl cholinesterase (AchE) were measured using commercial kits. qRT-PCR was used to detect NLRP3, Caspase-1 mRNA expression; Western blotting was used to detect NLRP3/Caspase-1 signal pathway related protein expressions. The results showed that, compared to the control group, the model group had less obvious structural hierarchy, sparser cell arrangement, wrinkled cytosol morphology, increased expressions of IL-6, TNF-α, AchE, increased mRNA and protein levels of NLRP3 and Caspase-1, and decreased Ach level (P<0.05). Compared to those of the model group, the cellular structural hierarchy and morphology in the low-dose CTS group, the medium-dose CTS group, and the high-dose CTS group all showed  dose-dependent improvement, as well as decreased expressions of IL-6, TNF-α, AchE, mRNA and protein levels of NLRP3 and Caspase-1 and increased Ach (P<0.05). Compared to the high-dose CTS group, the activator group showed aggravated cellular structural hierarchy and morphological changes, increased expression levels of IL-6, TNF-α, AchE, mRNA and protein levels of NLRP3 and Caspase-1, and decreased Ach (P<0.05). This study suggests that CTS may ameliorate neuroinflammation in SP rats by inhibiting the NLRP3/Caspase-1 signaling pathway.

To study the effect of ligustrazine on intestinal flora in atherosclerosis (AS) rats by regulating the TLR4/myeloid differentiation factor 88 (MyD88)/ NF-κB signaling pathway, SD rats were randomly separated into the control group, the model group, the low-dose ligustrazine group the high-dose ligustrazine group, and the high-dose ligustrazine+LPS group. The AS model was established by feeding high-fat diet combined with vitamin D3 injection. After intervention with ligustrazine and TLR4 activator LPS, body weight and the levels of low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), total cholesterol (TC), and triglyceride (TG) in blood lipids were measured. H-E staining was used to detect the morphology of thoracic aorta, liver and kidney tissues. Oil Red O staining was used to detect the plaque formation of thoracic aorta tissue. Myeloperoxidase (MPO) immunofluorescence staining was used to detect inflammatory cell infiltration of thoracic aorta tissue. 16S rRNA sequencing was used to detect changes in gut microbiota. ELISA was applied to determine the levels of serum endothelin-1 (ET-1), ICAM-1, GM-CSF and TNF-α. Western blotting was used to detect the expressions of TLR4/MyD88/NF-κB signaling pathway related proteins in thoracic aortic tissue. The results showed that compared to those of the control group, the LDL-C, TG and TC levels, proportion of thoracic aortic plaque area and MPO positivity ratio, Ace and Chao1 indexes, relative abundance of Proteobacteria, Escherichia and Coprococcus, ET-1, ICAM-1, GM-CSF and TNF-α levels, TLR4 and MyD88 protein expressions, p-NF-κB p65/NF-κB p65 of the model group were significantly increased (P<0.05), while the HDL-C level, Shannon index, relative abundance of Bacteroidetes and Lactobacillus were significantly reduced (P<0.05). Ligustrazine could improve the aforementioned pathological changes in AS model rats, with a stronger effect at higher dosage. LPS reversed the improvement effect of ligustrazine on the aforementioned pathological changes in AS model rats. This study suggests that ligustrazine improves gut microbiota composition in AS rats by reducing the activity of the the TLR4/MyD88/NF-κB signaling pathway, thereby alleviating arterial tissue damage.

Rheumatoid arthritis (RA) is a chronic autoimmune disease that can lead to joint deformity and loss of function. Due to the complexity of pathological mechanism and diversity of inflammatory cytokines, current treatments still lacks clinical efficacy and new treatment strategies have been continuously explored internationally. There is evidence suggesting that exosome and the miRNA encapsulated in them are involved in the pathogenesis of RA. Exosomal miRNA show great potential as biomarkers for the diagnosis of RA and as diagnostic tools for accurate identification and target therapy. As the most promising drug delivery vector, exosomes are expected to become one of the new treatment methods for RA. In recent years, researchers have conducted in-depth studies on the role of exosomal miRNA in the pathogenesis and progression of RA. This review summarizes the research progress on the mechanism of exosomal miRNA involved in RA pathogenesis and explores their potential clinical application prospect.

The aim of the study is to investigate the expression of serum TLR4 and IL-33 in patients with immune thrombocytopenia (ITP) and its clinical significance. Two hundred and twenty four ITP patients were enrolled as the study group and divided into the severe group (platelet count [PLT] <10×10⁹/L and presence of bleeding symptoms requiring treatment) and the non-severe group (PLT<30×10⁹/L, without bleeding symptoms) according to the severity of the patients' condition. The ITP patients were divided into the treatment ineffective/relapse group and the treatment effective group based on the follow-up of the patients' prognosis. Another 224 cases of healthy physical examinees during the same period were selected as the control group. The levels of serum TLR4 and IL-33 were analyzed by ELISA. Pearson correlation analysis was used to analyze the relationship between serum TLR4, IL-33 and mean platelet volume (MPV), PLT. Logistic regression analysis was used to analyze the influencing factors of the ITP ineffective/relapse group. The diagnostic and predictive value of serum TLR4 and IL-33 in patients with severe ITP or in ineffective/relapse patients were analyzed by ROC curve. The results showed that serum levels of TLR4, IL-33, and MPV in the ITP group were significantly higher than those in the control group (all with P<0.001). The serum levels of TLR4, IL-33, and MPV increased significantly in severe groups compared to those in the non-severe group (all with P<0.001). Serum TLR4 was positively correlated with IL-33（r=0.513, P<0.001）, and both serum TLR4 and IL-33 levels were positively correlated with MPV (r=0.512, r=0.516, both P<0.001). The AUC of the combined diagnosis by serum TLR4 and IL-33 for severe ITP was 0.848, more accurate than diagnosis by TLR4 or IL-33 alone  (both P<0.05). Compared to those of the effective treatment group, the serum levels of TLR4, IL-33, and MPV were increased (all with P<0.001) in the ineffective/relapse treatment group. TLR4, IL-33, and MPV were the risk factors for the ITP ineffective/relapse group (all with P<0.05), and PLT was a protective factor (P<0.05). The combined prediction of serum TLR4 and IL-33 for the ineffective/relapse group of ITP patients showed AUC of 0.976, and the combined prediction was superior to each of the individual predictions (all with P<0.05). In conclusion, the serum levels of TLR4 and IL-33 in ITP patients with are significantly increased, and correlate with severity of the disease condition and patient prognosis. 

This study investigates the effects of sevoflurane on the growth of colon cancer cell line CT26 and the M1 polarization of macrophages. RAW264.7 macrophages were divided into the negative control group, the model group (LPS-induced M1 polarization), the sevoflurane groups of different concentrations (1%, 2%, 4%), and the 2% sevoflurane+2-deoxy-D-glucose (2-DG; the glycolysis inhibitor) group. Conditioned media from these macrophages culture was co-cultured with CT26 cells, and divided into the control group (CT26 alone), the model group (CT26 with media from LPS-treated macrophages), the sevoflurane groups of different concentrations (CT26 co-cultured with macrophage media with 1%, 2%, 4% sevoflurane treatment), and the 2% sevoflurane + 2-DG group (CT26 co-cultured with conditioned macrophage media treated with 2% sevoflurane + 2-DG). A mouse tumor xenograft model was established and divided into the model group, the 2% sevoflurane group, the 2% sevoflurane + 2-DG group, and the macrophage-depleted + 2% sevoflurane group. Western blotting was used to detect the expressions of CD86, inducible nitric oxide synthase (iNOS), and glycolysis marker proteins including pyruvate dehydrogenase kinase 1 (PDK-1), hexokinase 2 (HK2), lactate dehydrogenase A (LDHA), and hypoxia-inducible factor 1α (HIF-1α). Levels of TNF-α, IL-6, and IL-12 were measured by ELISA. The results showed that compared to that in the control group, the proliferation ability of CT26 cells was inhibited in the sevoflurane group. Sevoflurane treatment increased the proportion of M1 macrophages and the expressions of glycolysis marker proteins. Conditioned media from sevoflurane-treated macrophages inhibited CT26 cell proliferation and colony formation, and the effect was attenuated by 2-DG. In the mouse tumor xenograft model, 2% sevoflurane reduced tumor volume and increased the proportion of M1 macrophages, glycolysis protein levels, and inflammatory cytokine levels in tumor tissue, which were counteracted by 2-DG. Depleting macrophages weakened the anti-tumor effect of sevoflurane compared to the 2% sevoflurane group. In conclusion, sevoflurane may inhibit colon cancer cell growth by modulating macrophage glycolysis to promote M1 polarization.

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by an increase in auto-antibodies, with complex clinical manifestations and limited therapeutic efficacy. Efferocytosis, the physiological process of apoptotic cell clearance, plays a pivotal role in maintaining immune homeostasis. Defective efferocytosis is associated with the pathogenesis of SLE by exposing self-antigens, triggering inflammatory responses, and promoting auto-antibody production. The molecular mechanisms of SLE-associated efferocytosis impairments include elevated “find-me” signals, aberrant expression and distribution of phosphatidylserine (“eat-me” signals) and its receptors, formation of antibodies against phosphatidylserine and its receptors, upregulation of “don't-eat-me” signals, and deficiencies in key enzymes or receptors essential for phagocytic engulfment and digestion. This review concludes current advances on the pathogenic role of efferocytosis defects in SLE and the underlying molecular pathways, providing new directions for future clinical diagnosis and drug development of SLE.

To facilitate the prompt identification of thyroid cancer lymph node metastasis during surgery, this study developed a method for the quick and accurate determination of thyroid globulin (Tg) content in intraoperative cervical lymph node puncture fluid using fine-needle aspiration (FNA) combined with lateral flow time-resolved immunofluorescence assay. A time-resolved fluorescence immunochromatographic assay for Tg in tissue fluid was established, and the performance was validated. The optimal intraoperative sampling method for cervical lymph node tissue fluid was established, and reference values were determined based on pathological results to verify clinical feasibility. The developed rapid Tg detection method demonstrated excellent stability and precision within a 15-minute detection window, with intra-assay and inter-assay coefficients of variation at 8.38% and 11.24%, respectively. It exhibited high sensitivity, detecting concentrations as low as 0.02 ng/mL, and high specificity without cross-reactivity with TPO. The sampling protocol involved 6 punctures in all directions of the lymph node using a 26-gauge needle, with repeated aspiration and injection 3 times into the sample preservation solution. Using 7.36 ng/mL as the reference value and comparing it with pathological results, the clinical positive coincidence rate was 95.16%, and the negative coincidence rate was 96.47%. The above results indicate that the rapid Tg detection method via FNA of lymph node tissue fluid is clinically feasible and effective for identifying the nature of thyroid cancer lymph node metastasis during surgery.

This study aims to investigate the effect of dihydroorotate dehydrogenase (DHODH) expression on metastasis and angiogenesis of non-small cell lung cancer (NSCLC) and the underlying molecular mechanism. Pathological tissues resected through surgery from 40 NSCLC patients were collected. Immunohistochemical staining was used to detect DHODH expression in tumor and adjacent normal tissues. The NSCLC cell lines A549, H1299, and H460, as well as a normal bronchial epithelial cell line 16HBE, were cultured, and the expression levels of DHODH were detected by Western blotting. The A549 cells were divided into the DHODH-OE-A549 and the DHODH-OENC-A549 groups, while the H1299 cells were divided into the shDHODH-H1299 and the shDHODH-NC-H1299 groups. Western blotting was performed to detect the expressions of TGF-β1/SMAD3 pathway related proteins, N-cadherin, E-cadherin, vimentin, and zonula occludens 1 (ZO-1). The wound-healing assay was used to assess cell migration, and the Transwell assay was applied to evaluate cell invasion. The cell culture supernatants were co-cultured with human umbilical vein endothelial cells (HUVEC) to observe changes in angiogenesis. The results showed that DHODH expression was significantly higher in NSCLC tissues than in adjacent normal tissues. Compared to that of 16HBE cells, DHODH expression levels were markedly elevated in A549, H1299, and H460 cell lines (all with P＜0.05). Compared to DHODH-OENC-A549 cells, DHODH-OE-A549 cells exhibited significantly increased expression of TGF-β1, p-SMAD3, vimentin, and N-cadherin, while SMAD3, E-cadherin, and  ZO-1 expression levels decreased. Cell migration, invasion, and angiogenic capacities were all significantly enhanced (all with P＜0.05). Conversely, compared to shDHODH-NC-H1299 cells, shDHODH-H1299 cells showed decreased expressions of TGF-β1, p-SMAD3, vimentin, and N-cadherin, but increased expressions of SMAD3, E-cadherin, and ZO-1, accompanied by significantly suppressed migration, invasion, and angiogenic activities (all with P＜0.05). In conclusion, DHODH expression is up-regulated in NSCLC and influences tumor metastasis and angiogenesis, which may due to the activation of the TGF-β1/SMAD3 signaling pathway.

he purpose of this study is to investigate the distribution of allergens in children with positive serum specific IgE (sIgE) for allergens in the western part of Shenzhen based on the data from the Shenzhen Baoan women's and children's Hospital. A retrospective analysis was conducted on allergen detection data and related clinical records. The results showed that with increasing age, both the types and numbers of positive allergen tests in children showed a linear upward trend, and the number of multiple positive allergen-sIgE (≥2) also increased linearly.  The top five  inhalant allergens were Dermatophagoides pteronyssinus (27.87%), Dermatophagoides farinae (26.85%), dust mite (26.68%), cat dander (6.41%), and Blomia tropicalis (5.86%). The positive rates were significantly higher in male children than in female children (P< 0.05).The top five food allergens were cow's milk (17.50%), egg white (15.28%), cashew (8.24%), crab (4.76%), and  shrimp (3.59%). Except for egg white,  sIgE positivity to the other four allergens was significantly higher in male children than in females (all P<0.05). The sIgE positivity rates for milk and egg  white decreased progressively with age (P<0.001). Sensitization to Dermatophagoides pteronyssinus, Dermatophagoides farinae, dust mite, and cat dander peaked during summer months, whereas Blomia tropicalis sensitization reached its highest level in spring, with all seasonal variations being statistically significant (P<0.001).  The level of sIgE positivity is related to the age and gender of the children. Overall, the sIgE positivity rate is higher in male than in female children, and with increasing age, the spectrum of sIgE-positive allergens expands significantly, and the proportion of multiple positivity shows a linear increase.

To investigate the expression of 7-dehydrocholesterol reductase (DHCR7) in microsatellite-instability (MSI) gastric cancer tissues and its correlation with the immune microenvironment and prognosis, stratified regression analysis, GAM analysis, and Cox regression analysis were used to retrospectively analyze the DHCR7 expression level and immune microenvironment data of 281 gastric cancer patients admitted from July 2020 to May 2022. The results showed that in microsatellite-stable (MSS) gastric cancer tissues, the rate of elevated expression of DHCR7 protein was 68.25%, which was significantly higher than that in the adjacent normal tissue (10.58%; χ²=21.143, P<0.05). In MSI gastric cancer tissues, the rate of DHCR7 high-expression was 57.61%, also significantly higher than that of the adjacent normal tissues (8.70%; χ²=25.329, P<0.05). In MSI gastric cancer tissue, the expression of DHCR7 protein correlated with distant metastasis, TNM stage, lymph node metastasis, and degree of tissue differentiation (P<0.05). In the high-DHCR7-expression patients, memory resting CD4⁺T cells were significantly lower than those of the low-expression group, while indicators such as M0 macrophages, M1 macrophages, and M2 macrophages were significantly higher (P<0.05). Stratified regression analysis showed that lymph node metastasis and TNM stage positively correlated with the level of DHCR7 mRNA, and the degree of tissue differentiation had a negative correlation (P<0.05). The factors related to the immune microenvironment had a statistically significant impact on the expression of DHCR7  mRNA (P<0.05). Multivariate Cox regression analysis indicated that multiple factors, such as lymph node metastasis, were independent risk factors affecting the postoperative progression-free survival (PFS) and overall survival (OS) of MSI gastric cancer patients (P<0.05). The ROC curve showed that the level of DHCR7  mRNA may predict the 2-year PFS and 2-year OS of MSI gastric cancer patients to a certain extent. Restricted cubic spline and threshold effect analysis showed a non-linear positive correlation between the expression level of DHCR7 and the prognosis of MSI gastric cancer patients. The 2-year PFS and 2-year OS of patients in the high-DHCR7-expression group were lower than those in the low-expression group (P<0.05). This study suggests that in MSI gastric cancer tissues, DHCR7 is highly expressed, which affects the prognosis of MSI gastric cancer patients and the tumor immune microenvironment.

In this study, a widely targeted metabolomics (liquid chromatograph-mass spectrometer ［LC-MS］) approach was used to qualitatively and quantitatively analyze the gut microbiota metabolites between individuals with irritable bowel syndrome (IBS) and healthy controls and to compare the differential metabolites between the two groups. 30 patients with irritable bowel syndrome (IBS) admitted to PLA Naval Medical Center from January 2023 to September 2023 were selected as the study subjects. After exclusion based on inclusion/exclusion criteria, 8 remaining cases were assigned to the IBS group, while 8 healthy individuals matched by sex and age during the same period served as the control group. Fecal samples were frozen at  immediately after collection, and IL-1β, IL-6, and TNF-α were detected by ELISA. LC-MS was used to analyze the difference and enrichment of the extracts. The results showed that the levels of IL-1β (P<0.001), IL-6 (P<0.001), and TNF-α (P<0.05) in the IBS group were significantly higher than those in the control group. LC-MS analysis identified 46 different metabolites between the IBS group and the control group. These metabolites included 10 classes, mainly amino acids and their derivatives, nucleotides and their derivatives, organic acids and their derivatives, lipids, flavonoids, lignans and coumarins, alkaloids, vitamins and their derivatives, and steroid hormones, accounting for 87% of all metabolites detected. Further hierarchical cluster analysis and principal component analysis (PCA) demonstrated clear separation between the two sample groups. Screening of differential metabolites and pathway analysis revealed that these metabolites were primarily enriched in amino acid-related pathways, including valine, leucine, and isoleucine biosynthesis and degradation, aminoacyl-tRNA biosynthesis, and tryptophan metabolism pathways. Amino acids and their derivatives, such as valine, leucine, tryptophan, arginine, and methionine, are key metabolites for antioxidation and inhibition of inflammatory response, and may be involved in the occurrence and development of psychological stress symptoms such as insomnia and anxiety. Therefore, quantitative evaluation of differential metabolites and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis are helpful to explore the pathogenesis of IBS from the perspective of amino acid metabolism, and lay a foundation for preventing and treating physiological and psychological stress of soldiers.