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.

The objective of this study was to verify the performance of time-resolved fluorescence immunoassay for human serum anti-phospholipase A2 receptor antibody IgG4 (PLA2R-IgG4) including the linear range, accuracy, precision, specificity, and performance of PLA2R-IgG4 reagent with magnetic particle time-resolved fluorescence immunoassay. The results showed that the blank limitation was 7.02 ng/mL, linear range was 50.00 ng/mL-10 000.0 ng/mL, accuracy was 86.28%-104.58%, intra-batch precision was 2.62%-5.33%, inter-batch precision was 7.37%-8.26%, and the specificity was in acceptable range. The detection rate of PLA2R-IgG4 in 77 idiopathic membranous nephropathy patients was 75.32%, and the test results of 15 patients with secondary membranous nephropathy were all negative. All the indexes of this method meet the requirements of clinical test quality, and this method shows a better detection rate for idiopathic membranous nephropathy, making it promising in clinical translation. 

Obesity is the excessive accumulation of fat caused by the imbalance between energy intake and expenditure, which has become a global health issue. The metabolic homeostasis of adipose tissue plays a crucial role in the progression of obesity and has numerous impacts on systemic physiology. Studies have found that various immune cells and cytokines are involved in maintaining and regulating the metabolic homeostasis of adipose tissue, suggesting the significant potential of immune cells in alleviating obesity and insulin resistance (IR). From the perspectives of innate and adaptive immunity, this review discusses the progress on the role of immune cells in thermogenesis and metabolic functions of adipose tissue and further explores the therapeutic potential of the immune system in obesity and related metabolic disorders.

The aim of this study is to explore the regulatory features, co-regulatory roles and effects on the epigenome of the STAT family proteins and the master transcription factors of the Th1, Th2, and Th17 subtypes, including the T-box expressed in T cell (T-bet), GATA binding protein 3 (GATA3), and retinoic acid receptor-related orphan receptor γt (RORγt) in Th differentiation. Assay for transposase-accessible chromatin using sequencing (ATAC-seq) data of different T cell differentiation stages and chromatin immunoprecipitation sequencing (ChIP-seq) data of transcription factors were obtained and integrated from the GEO database. The co-localization, the genomic distribution of the transcription factors, and the chromatin accessibility changes during differentiation and functions of regulated genes were analyzed. The results showed that the transcription factor STAT2 was the main STAT family transcription factor involved in the maintenance of Th0 chromatin characteristics. STAT4 was widely co-localized with T-bet and regulated Th1 activation. The master regulator GATA3 and the co-factor STAT6 preferentially bound to non-promoter cis-elements thereby directing Th2 differentiation. RORγt, the master transcription factor, enhanced the binding strength of STAT3 and STAT5 in a distinctive manner independent of altered chromatin accessibility, co-regulating Th17-specific differentiation. This study suggests that STAT family transcription factors selectively participate in and differentially regulate Th differentiation.

The purpose of this study is to explore the role and mechanism of peptidyl arginine deimidase type 4 (PAD4) in inflammatory adipose tissue. The subcutaneous and visceral adipose tissues of db/db (type 2 diabetes mellitus [T2DM] group), db/m (T2DM control group), ob/ob (obese group) and ob/c(obese control group) were extracted, and the expression of PAD4 in subcutaneous and visceral adipose tissue of T2DM and obese mice was detected by immunohistochemistry. Wild-type mouse primary peritoneal macrophages and mouse macrophage cell line RAW264.7 were stimulated with LPS, the expressions of PAD4, inflammatory factors TNF-α and IL-6, the activation of NF-κB signal pathway in the 2 types of cells were detected by qRT-PCR and Western blotting, respectively. The results showed that compared to those of the db/m and ob/c groups, the expression of PAD4 in subcutaneous and visceral adipose tissue, primary peritoneal macrophages and RAW264.7 cells stimulated by LPS in db/db and ob/ob groups were all decreased, while the expressions of inflammatory factors TNF-α and IL-6 increased significantly.Meanwhile, the TLR4/NF-κB signal pathway was activated. This study shows that the expression of PAD4 is decreased in subcutaneous and visceral adipose tissue in inflammatory state (T2DM and obesity) and is negatively correlated with the activation of the TLR4/NF-κB signal pathway.

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.

To investigate the role of HS1-associated protein X-1 (HAX-1) in lupus nephritis (LN) patients, 50 patients with SLE were selected as the study objects, and 29 healthy patients were selected as the control group. qRT-PCR was used to detect the mRNA expression of HAX-1  in PBMC. The expression of HAX-1 in kidney tissue was detected by immunohistochemistry (IHC). The proportions of CD4⁺ CD25⁺ Foxp3⁺ Treg and CD4⁺ IL-17A⁺ Th17 in peripheral blood were measured by flow cytometry. Human renal mesangial cells (HRMC) were cultured in vitro, and HAX-1 silencing plasmid and HAX-1 overexpression plasmid were transfected. Flow cytometry and MTT were used to measure the apoptosis and proliferation of HRMC after transfection, respectively. Western blotting was used to detect the expressions of proliferation-related signal PI3K/Akt and apoptosis-related signal Bax/Casepase-9. The results showed that the mRNA expression level of HAX-1  in PBMC of the active LN group was significantly higher than that of the NRA-SLE group (P<0.001). Meanwhile, HAX-1  mRNA  expression level was positively correlated with SLE disease activity index (SLEDAI-2K) and 24 h urinary protein quantity in peripheral blood (R2= 0.876, P<0.001; R2=0.409, P<0.001). The IHC results showed that the expression level of renal HAX-1 in the active LN group was significantly higher than that of the normal control group (P<0.001). The expression level of HAX-1 in renal tissue was positively correlated with LN activity score and chronicity score (r=0.975, P<0.001; r=0.667, P=0.002). Overexpression of HAX-1 reduced apoptosis (P<0.001) and increased the proliferation activity of HRMC (P<0.001). Meanwhile, the levels of PI3K and p-Akt significantly increased (P<0.001) while the levels of Bax/Casepase-9 significantly decreased (P<0.001). In contrast, upon transfection of HAX-1 silencing plasmid, the apoptosis of HRMC was significantly increased (P<0.001), and the proliferation activity was significantly reduced (P<0.001), along with significantly reduced PI3K and p-Akt expression (P<0.001), increased Bax/Casepase-9 expression(P<0.001). These results suggest that HAX-1 expression in peripheral blood and kidney tissues of patients with active LN group is closely related to kidney involvement and disease activity, as well as the imbalance of the CD4⁺ IL-17A⁺ Th17 and CD4⁺ CD25⁺ Foxp3⁺ Treg. Using a system of HRMC cultured in vitro, this study demonstrates that HAX-1 can promote the pathogenesis  of glomerular nephritis in LN by affecting the signaling pathways related to mesangial cell proliferation and apoptosis. HAX-1 may have a dual effect on Th17/Treg balance and HRMC proliferation, contributing to the pathogenesis of LN. 

This study aimed to evaluate the performance and property of a fully automated ultra-high speed immunoassay system based on high-throughput flow fluorescence technology for detecting autoimmune antibodies in clinical laboratories. According to the laboratory accreditation guidelines issued and implemented by the China National Accreditation Commission for Conformity Assessment (CNAS), flow fluorescence technology was used to detect the autoantibodies and evaluations  were made on parameters including the precision, linear range, accuracy, and limit of blank (LOB). When conducting repeatability validation on quality control products at both low and high concentrations, the average standard deviation and coefficient of variation (CV)  were calculated of each antibody level in the 16 antibody profiles. The CV of all antibodies were lower than 7% and 5%, respectively, which were within the less than 10% range as declared by the manufacturer. The first-order polynomial linear regression coefficients for resistance to anti-C1q-Ab and resistance to anti-dsDNA-Ab were 0.999 and 0.996, respectively, with correlation coefficient  above 0.999. The deviation distributions were all within 3% and the results of the 2 recovery experiments were 91.56% and 105.60%, respectively. When Oumeng company imprinting method was used as a reference, the compliance rates of the validation projects were all above 92%. Values of LoB for anti-C1q-Ab and anti-dsDNA-Ab were both within 0.5 U/ml and 3 IU/ml, respectively. In conclusion, all performance parameters of the flow cytometry fluorescence technology for autoantibody detection are consistent with the manufacturer's specifications and pass the standard of clinical requirements. It can replace similar imported instruments.

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 aims to investigate the regulatory effect of glutathione S-transferase mu 3 (GSTM3) on renal tubular epithelial cells HK-2 pyroptosis and renal inflammation in hyperuricemia (HUA). GSE186871 data set was used to screen for kidney differentially expressed genes in HUA and normal mice. The mice were divided into the normal group, the model group, the normal + adenovirus shRNA (normal + Ad-sh-NC) group, the model+Ad-sh-NC group (Ad-sh-NC group), and adenovirus GSTM3 shRNA group (Ad-sh-GSTM3 group）. HK-2 cells were divided into the control group, the model group, the control + Ad-sh-NC group, the Ad-sh-NC group, and the Ad-sh-GSTM3 group. Cell survival rate was measured by CCK-8 method. Renal pathology was observed by H-E staining. The levels of serum uric acid (UA), creatinine (Cr)，and blood urea nitrogen (BUN) were detected by a biochemical analyzer. IL-1β and IL-18 levels were detected by ELISA. Western blotting was used to detect GSTM3, Caspase-1, NOD-like receptor family pyrin domain-containing protein 3 (NLRP3), gasdermin D (GSDMD)-N, and Cleaved GSDMD-N expressions. The expressions of GSTM3 and GSDMD-N in kidney were detected by immunohistochemistry. The results showed that compared to the normal group, the model group showed renal tubule swelling and local inflammatory cell infiltration. The expressions of GSTM3, Caspase-1, NLRP3, GSDMD-N, and Cleaved GSDMD-N in renal tissue, and serum levels of Cr, BUN, UA, IL-1β, and IL-18 were all increased (P<0.05). Compared to the model group, the Ad-sh-GSTM3 group exhibited reduced renal tubule injury, decreased levels of Cr, BUN, UA, IL-1β, and IL-18 (P<0.01), and decreased protein expressions of GSTM3, Caspase-1, NLRP3, GSDMD-N, and Cleaved GSDMD-N (P<0.05). Compared to those of the control group, the HK-2 cell survival rate in the model group was decreased (P<0.01), along with increased IL-1β and IL-18 levels in the supernatant (P<0.01), and increased protein expressions of GSTM3, Caspase-1, NLRP3, GSDMD-N, and Cleaved GSDMD-N (P<0.01). Compared to the model group, the Ad-sh-GSTM3 group showed increased HK-2 cell survival rate (P<0.01), decreased IL-1β and IL-18 levels (P<0.01), and decreased protein expressions of GSTM3, Caspase-1, NLRP3, GSDMD-N, and Cleaved GSDMD-N (P<0.01). Therefore, inhibition of GSTM3 effectively inhibits the pyroptosis of renal tubular epithelial cells and alleviate the renal inflammatory injury caused by HUA.

This study was designed to explore the potential relationship between myocardial fibrosis and the pyroptosis of mouse cardiac fibroblast (MCF) regulated by soluble growth stimulation expressed gene 2 (sST2). A mouse model of viral induced myocarditis (VMC) was established, and mice were divided into the VMC group and the VMC+ST2 antibody group in addition to a control group. Mice were sacrificed on the 7th day and cardiac inflammatory infiltration was observed by H-E staining and collagen deposition was observed by Masson staining. Western blotting was conducted to detect the expressions of NOD-like receptor family pyrin domain-containing protein 3 (NLRP3), IL-1β, cleaved caspase-1, GSDMD-N, and activation markers collagen Ⅰ/Ⅲ and α-SMA after sST2 treatment of MCF. After MCC950 pretreatment, the expressions of NLRP3, IL-1β, cleaved caspase-1, GSDMD-N, and activation markers collagen-Ⅰ/Ⅲ and α-SMA were detected. The mRNA expressions of IL-1β, IL-18, NLRP3, collagen-Ⅰ/Ⅲ and α-SMA after MCC950 pretreatment were detected by qRT-PCR. The results showed that the expressions of NLRP3, IL-1β, IL-18, cleaved caspase-1, and GSDMD-N were significantly increased after sST2  treatment of MCF. Activation indexes collagen-Ⅰ/Ⅲ and α-SMA were up-regulated. After MCC950 pretreatment, the expressions of NLRP3, IL-1β, IL-18, cleaved caspase-1, and GSDMD-N were down-regulated. The activation indexes of collagen-I/III and α-SMA were significantly reduced. In vivo ST2 blockage caused reduced myocardial inflammatory infiltration and collagen deposition. In conclusion, sST2 activates NLRP3 inflammasome to induce MCF pyroptosis and promote myocardial fibrosis.

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.

To explore the protective effect of miR-34b-5p on sepsis induced acute lung injury (ALI) in rats and its impact on the phosphatase and tensin homolog (PTEN)/Akt/mTOR pathway, 80 specific pathogen free grade male SD rats were randomly divided into the sham group, the model group, the empty vector group, and the miR-34b-5p group, with 20 rats in each group. The rats in the sham group only underwent laparotomy, while the remaining rats were subjected to cecal ligation and perforation to establish a sepsis model. After the establishment of the model, rats in the empty vector group and the miR-34b-5p group were injected with empty vectors and miR-34b-5p overexpression plasmids through the tail vein, respectively. Rats in the model group and the sham group were injected with equal amount of physiological saline. Three days after injection, ELISA was used to determine the IL-1β and IL-6 levels in rat peripheral blood. The wet weight/dry weight  (W/D）ratio of rat lung tissue was calculated and the pathological changes of rat lung tissue was observed by H-E staining. The apoptosis of lung histiocyte cells was detected by TUNEL method. miR-34b-5p, IL-1β and IL-6 mRNA levels in lung tissue were detected by qRT-PCR. Western blotting was used to detect the protein levels of PTEN, p-Akt, p-mTOR, Akt, mTOR, B cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-3. The results showed that the alveoli in the sham group appeared normal, while those of the model group and the empty vector group were collapsed, along with thickened alveolar walls, vascular congestion, pulmonary interstitial congestion, edema, and inflammatory cell infiltration. The infiltration of inflammatory cells in the lung tissue of rats in the miR-34b-5p group was reduced, and alveolar injury was significantly improved. Compared to those of the sham group, the expressions of IL-1β and IL-6 at both protein and mRNA levels, the number of apoptotic cells, levels of Bax, Caspase-3 and PTEN as well as W/D ratio were significantly increased in the model group (P<0.01), while the levels of miR-34b-5p, Bcl-2 protein, p-Akt/Akt ratio, and p-mTOR/mTOR ratio were significantly decreased (P<0.01). Compared to those of the empty vector group, the miR-34b-5p expression, Bcl-2 protein level, p-Akt/Akt and p-mTOR/mTOR ratio in the lung tissue of the miR-34b-5p group were significantly increased (P<0.01), while the IL-6, IL-1β protein and mRNA levels, the number of apoptotic cells, the Bax, PTEN, Caspase-3 protein levels, and W/D ratio were significantly decreased (P<0.01). This study suggests that miR-34b-5p alleviates pulmonary inflammation and cell apoptosis of ALI in sepsis rats through the PTEN/Akt/mTOR pathway.

To analyze the effect of total alkaloids of Strychni Semen on the apoptosis of synovial cell and on the Wnt3a/β-catenin signaling pathway in rheumatoid arthritis (RA) rat, a total of 40 SPF-level SD rats were prepared, with 10 for the control group and 30 for the RA model group. Twenty-seven rats in total were successfully modeled and were divided into the model group, the total alkaloids of Strychni Semen group, and the total alkaloids of Strychni Semen+Wnt3a agonist group, with 9 rats per group. Histopathological features, toe-swelling degree, arthritis index, immune organ index, synovial cell apoptosis rate, inflammatory factors concentration, and Wnt3a/β-catenin pathway gene transcription and expression were measured in each group. The results showed that the synovial tissue structure of the control group was normal without hyperplasia or inflammatory infiltration. In the model group, the synovial tissue shape was rough, with hyperplasia and infiltration of inflammatory cells. The synovial hyperplasia and inflammatory infiltration were alleviated in the total alkaloids of Strychni Semen and the total alkaloids of Strychni Semen+Wnt3a agonist groups. Compared to those of the control group, toe swelling, arthritis index, thymus index, spleen index, IL-6, IL-17, IL-1β, and TNF-α levels, Wnt3a, β-catenin mRNA and protein expression levels increased in the model group, while the apoptosis rate of synovial cells decreased. Compared to those of the model group, toe swelling, arthritis index, thymus index, spleen index, IL-6, IL-17, IL-1β, and TNF-α levels, Wnt3a, β-catenin mRNA and protein expression in the total alkaloids of Strychni Semen group were decreased, while the apoptosis rate was increased. Compared to the total alkaloids of Strychni Semen group, the total alkaloids of Strychni Semen+Wnt3a activator group showed increased toe swelling, arthritis index, thymus index, spleen index, IL-6, IL-17, IL-1β, and TNF-α concentrations, Wnt3a, β-catenin mRNA and protein expression levels and decreased apoptosis rate of synovial cells (all above with P＜0.05). In conclusion, total alkaloids of Strychni Semen can inhibit intraarticular inflammation and promote synovial cell apoptosis in RA rats by modulating the Wnt3a/β-catenin signaling pathway.

As a key regulator of immune responses, macrophages can be polarized into types M1 and M2 and affect the onset, progression, and treatment of cardiovascular diseases. M1 macrophages, characterized by their pro-inflammatory properties, can exacerbate inflammatory responses and thereby accelerate the development of cardiovascular diseases. In contrast, M2 macrophages, with their anti-inflammatory characteristics, contribute to the recovery from cardiovascular diseases. This review summarizes the correlation between M1/M2 macrophage subtypes and cardiovascular diseases including atherosclerosis (AS), essential hypertension (EH), and myocardial infarction (MI), as well as potential regulatory targets during the M1/M2 polarization process. This review provides a theoretical foundation and new research perspectives for further study and treatment of cardiovascular diseases.

The study aims to investigate the expression level of PD-1 on peripheral blood T cells among colon cancer patients with different stages and its connection with immunotherapy efficacy. One hundred and twelve colon cancer patients were enrolled in to analyze the relationship between PD-1 expression and clinical parameters. Based on the outcome of immunotherapy, the patients were divided into the effective group (n=85) and the ineffective group (n=27). Multivariate logistic regression formula was used to analyze the relationship between PD-1 expression and clinical characteristics and to screen the factors affecting the efficacy of immunotherapy in the objects. The results showed that there existed statistically significant differences in PD-1 expression among different clinical stages, and among different expression levels of carcinoembryonic antigen (CEA) or carbohydrate antigen 199 (CA199) (all with P＜0.05). However, the results of multivariate logistic analysis showed that the positive expression rate of PD-1 could not be used as an independent predictor for clinical staging (P＞0.05); Low/middle-low differentiation, clinical stage ⅢⅣ, lymph node metastasis, distant metastasis, neutrophil-to-lymphocyte ratio (NLR)≥5, platelet-to-lymphocyte ratio (PLR)≥135, and positive expression of PD-1 all were risk factors predicting ineffective immunotherapy in colon cancer patients (all with P＜0.05). This study suggests that the expression of PD-1 on CD4+T cells in peripheral blood is significantly associated with clinical stages and the efficacy of immunotherapy in colon cancer patients. However, it can not be used as an independent predictor for clinical staging. Meanwhile, PD-1 is sufficient to be used as a biological index for predicting the efficacy of immunotherapy.

This study aimed to investigate the effect of Bupiqiangli compound on Th17/Treg differentiation of thymic lymphocytes in rats with myasthenia gravis. CCK-8 was used to determine an optimal 10% serum concentration for subsequent experiments. Cells were divided into 8 groups: the normal group (control), the normal+blank serum group (control+BS), the myasthenia gravis model group (model), the myasthenia gravis model+blank serum group (model+BS), the myasthenia gravis model+low-dose Bupiqiangli compound treatment serum group (model+LBPQL), the myasthenia gravis model+medium-dose Bupiqiangli compound treatment serum group (model+MBPQL), the myasthenia gravis model+high-dose Bupiqiangli compound treatment serum group (model+HBPQL), and the myasthenia gravis model+prednisone containing serum group (model+prednisone). After treated with serum for 24 hours, the proportions of Th17 and Treg cells in each group were detected by flow cytometry. ELISA was used to detect the content of IL-17 and TGF-β in the supernatant. Western blotting was used to measure the protein levels of Notch1, Hes1, Hes5 and Hey1. The results showed that, compared to that of the spleen deficiency model group, the proportion of Th17 cells in the model+HBPQL group and the model+Prednisone group decreased significantly (P<0.05), while the proportion of Treg cells increased significantly (P<0.05). The expression of IL-17 was significantly decreased (P<0.05), while the expression of TGF-β was significantly increased (P<0.05). The protein expressions of Notch1, Hes1, Hes5, and Hey1 were significantly decreased (P<0.05). The results indicate that Bupiqiangli compound can regulate the proportions of Th17 and Treg in thymic lymphocytes in rats with myasthenia gravis. The underlying mechanism may involve the regulation of the Notch signaling pathway. 

This study aimed to investigate the role of IL-6 in diffuse large B-cell lymphoma (DLBCL) and the role of IL-6 on cell apoptosis, proliferation, invasion and migration via the JAK/STAT3 signaling pathway. OCI-LY8 cell line was used as a DCBCL model and drug intervention of IL-6 or STAT3 gene silencing were used to assess the effect of IL-6 and STAT3 on OCI-LY8 cells. Cell viability was detected by CCK8 assay and apoptosis rate was determined by flow cytometry to study the effect of IL-6 on the proliferation and apoptosis of DLBCL cells. Transwell assay and scratch assay were used to detect cell invasion and migration capacity and elucidate the effect of different IL-6 concentrations on DLBCL migration. The expressions of JAK, STAT3 and the downstream apoptosis-related B cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax) and survivin proteins were detected by Western blotting. Western blotting showed that IL-6 up-regulated anti-apoptotic proteins Bcl-2 and survivin, and downregulated pro-apoptotic protein Bax by activating the JAK/STAT3 pathway, thereby inhibiting OCI-LY8 apoptosis. IL-6 could also enhance the invasion and migration of OCI-LY8 by activating STAT3. The effect of IL-6 was reversed upon IL-6 inhibitor LMT-28 treatment or STAT3 silencing. IL-6 enhanced the invasion and migration capacity of OCI-LY8 DLBCL cells by activating the JAK/STAT3 signaling pathway, as well as increasing their anti-apoptosis ability. Together, these findings reveal that IL-6 promotes DLBCL development via the JAK/STAT3 pathway, which provides a new potential target and theoretical basis for DLBCL treatment.

PD-1 is a highly notable immune checkpoint molecule that plays an important role in tumor immune evasion. Tregs are a subset of T cells that negatively regulate the immune response, and are involved in the immune regulation of immune-related pathological processes or diseases such as tumor microenvironment, inflammatory microenvironment, autoimmune diseases, sepsis, and graft-versus-host disease (GVHD). Studies have shown that PD-1 is expressed not only on tumor cells but also on Tregs. This review summarizes the changes in PD-1 expression, the proportion of Tregs and the immune regulation of PD-1 on Tregs in immune-related pathological processes or diseases, aiming to provide new ideas for the study of these pathological processes or diseases.

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.

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.

The aim of this study is to understand the distribution of autoantibodies in systemic lupus erythematosus (SLE) patients and explore the role of SLE autoantibodies in organ function. Seventy-six newly diagnosed SLE patients admitted to the Second Hospital of Jiaxing  from January 2018 to December 2023 were selected. Indirect immunofluorescence was used to detect the autoantibodies index, and the data were analyzed. The results showed that most SLE patients suffered from arthritis (55.26%) and rash (52.63%). The positive rate of antinuclear antibody (ANA) was 100.00%, and among them the most common was the anti-Sjogren's syndrome A (SSA) antibody. Multiple autoantibodies in the vast majority of patients were positive (97.37%). The positive rate of anti-dsDNA antibody was the highest in the adolescent group (66.67%), but the lowest in the elderly group (33.33%). The positive rate of anti-dsDNA antibody in lupus nephritis (LN) was 76.47%. The positive rate of anti-Sm antibody was the highest in Raynaud＇s phenomenon and alopecia patients (33.33% and 50.00% respectively). The positive rate of anti-histone antibody (AHA) in vasculitis and LN patients was 75.00% and 58.82%, respectively. This study suggests that the clinical manifestations of SLE are different and highly heterogeneous, and multiple autoantibodies are correlated with organ involvement, clinical manifestations and age.

2025年8月29~30日，以“聚力同行，共创未来”为主题的2025年免疫学前沿进展学术年会在上海成功举办。本次年会由上海市免疫学会主办，其官方学术期刊《现代免疫学》杂志亦全程参与支持，汇聚了60余位国内外领军学者及超800名参会代表，共同探讨免疫学前沿发展趋势。

 年会特邀储以微教授、Tiffany Horng教授、郑利民教授、郑颂国教授及李华兵教授等知名专家作主旨报告，各位学者围绕各自研究领域的前沿突破展开深度分享，呈现了一场高水准的学术盛宴。会议同步设置皮肤免疫、眼免疫、神经免疫、移植免疫、交叉免疫、感染免疫、风湿免疫、中医药免疫八大专题分会场，有效推动基础研究、临床转化与生物技术等跨学科领域的深度碰撞与融合。 

本次年会不仅为免疫学界搭建了高水平的学术交流平台，更依托《现代免疫学》等专业期刊的学术传播优势，加速了科研成果向临床应用及产业转化的进程。此次盛会的成功举办，进一步凝聚了我国免疫学领域的创新力，为推动学科发展注入了新的动能。

The study aims to investigate the effect of IL-27 on multiple myeloma (MM) via the imbalance of Th17 and Treg. Peripheral blood samples from 10 MM patients and 10 healthy individuals were collected. qRT-PCR was used to detect IL-27, Th17-related factors (retinoic acid receptor-related orphan receptor gamma t ［RORγt］, IL-17A, and IL-6) and Treg-related factors (IL-10 and forkhead box protein 3 ［Foxp3］) in the peripheral blood. Western blotting was used to detect the key transcriptional pro-factors of Th17 and Treg. Flow cytometry was used to detect the changes in the ratios of Th17 and Treg in peripheral blood, and qRT-PCR was used to detect the cytokines secreted by Th17 (RORγt, IL-17A, IL-21, and IL-22) and the anti-inflammatory factors (Foxp3, IL-10, IL-35, and TGF-β) secreted by Treg in PBMC. RPMI-8226 cells were used to construct oe-NC/oe-IL-27 and si-NC/si-IL-27 cell lines. ELISA was used to detect the level of IL-27 in the supernatant of myeloma cells. Flow cytometry was used to detect the proportion of Th17 co-cultivated with CD4⁺ T cells in the overexpression cells. CCK-8 and Transwell assays were used to detect the cell viability, migration and invasion ability. The results showed that the levels of IL-27 and Treg-related factors (IL-10 and Foxp3) were increased (P<0.05) and Th17-related factors (RORγt, IL-17A, and IL-6) were decreased in the MM group (P<0.05). There was no difference in the proportion of Th17 between MM group and normal control group, while the proportion of Treg was increased in MM group. Levels of cytokines secreted by Th17 (RORγt, IL-17A, IL-21, and IL-22) were decreased (P<0.05) and anti-inflammatory factors secreted by Treg (Foxp3, IL-10, IL-35, and TGF-β) were increased in PBMC of MM patients (P<0.05). Myeloma cells could secrete IL-27. Th17-related factors (RORγt, IL-17A, IL-21, and IL-22) in the oe-IL-27 group decreased (P<0.05) while the cell viability increased (P<0.05). The above results of the oe-IL-27 group were reversed in the si-IL-27 group. This preliminary study confirms that IL-27 promotes MM progression via Th17/Treg imbalance.

To investigate the diagnostic value and biological functions of kinesin family member 11 (KIF11) in lung adenocarcinoma (LUAD) patients, this study utilized bioinformatics platforms and statistics (The Cancer Genome Atlas ［TCGA］, Human Protein Atlas ［HPA］, and ROC curve) to examine the expression of KIF11 in LUAD tissues and its relationship with patient survival, as well as the association between KIF11 and the clinicopathological features. Pearson correlation analysis was used to study the association between KIF11 expression and the 22 types of immune cells infiltration. qRT-PCR was used to detect the relative mRNA expression of KIF11 in LUAD cell lines. Western blotting was used to detect the expressions of KIF11 protein, epithelial-mesenchymal transition (EMT)-related proteins, and of cell cycle-related proteins. CCK8 assay, colony formation assay, Transwell assay, scratch healing assay, and flow cytometry were used to detect tumor cell proliferation, migration and invasion abilities, as well as cell cycle distribution, respectively. The results showed that KIF11 and  its protein were highly expressed in LUAD tissues and cell lines (all with P＜0.05), KIF11 level was closely connected with the patients clinical characteristics (all with P＜0.05), and the high expression of KIF11 led to reduced survival rate in patients (P＜0.05). Pearson correlation analysis revealed that the expression of KIF11 was positively correlated with the infiltration of the majority of immune cells (all with r＞0). Down-regulation of KIF11 expression inhibited the proliferation (P＜0.01), migration (P＜0.000 1) and invasion abilities (P＜0.01) of LUAD cells, increased the proportion of G2/M phase cells (P＜0.001), decreased the proportion of G0/G1 phase cells (P＜0.001), promoted apoptosis (P＜0.000 1), and inhibited the EMT process  (all with P＜0.05). This study demonstrates that KIF11 may be used as a diagnostic marker for LUAD, and down-regulation of the expression of KIF11 inhibits LUAD tumor cell progressive phenotypes.

The aim of this study was to investigate the therapeutic effect of acacetin on depressed rats and its regulation on the HIF-1α/NLRP3 signaling pathway. A rat model of depression was established, and rats were divided into the model group, the acacia low-dose group, the acacia high-dose group and the acacia high-dose + pathway activator group (acacia high-dose+BMS-986299 group), with 12 rats in each group, in addition to a control group. After the treatment was completed, the sugar water preference, the mine field experiment, and the forced exercise experiment were performed, respectively. ELISA was used to detect levels of inflammatory and oxidative stress factors. H-E and Nissl staining were used to observe the histopathological morphology and neuronal number of hippocampus. Western blotting was used to detect the expression of proteins related to the HIF-1α/NLRP3 signaling pathway. Compared to those of the control group, the hippocampal tissue structure of the model group was damaged, the pyramidal cell arrangement was disordered, the nuclear condensation phenomenon was obvious, and the numbers of Nissl bodies and neurons decreased. In the meantime, the sugar water preference rate, voluntary activity score, and the level of CAT were decreased, whereas the immobility time of forced swimming, the levels of IL-6, IL-β, TNF-α, reactive oxygen species (ROS), malondialdehyde (MDA), and the expression of HIF-1α, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and caspase-1 were increased (P<0.05). Compared to those of the model group, the hippocampal tissue structure of the acacia low-dose and high-dose groups was relatively normal, and the pyramidal cells were relatively intact and well arranged, the nuclear condensation phenomenon was reduced, and the numbers of Nissl bodies and neurons increased. Consistently, the sugar water preference rate, voluntary activity score, and the level of CAT increased, while the immobility time of forced swimming, the levels of IL-6, IL-β, TNF-α, ROS, MDA, and the expression of HIF-1α, NLRP3, ASC and caspase-1 decreased (P<0.05). Compared to those of the acacia high-dose group, the hippocampal tissue structure damage and pyramidal cell arrangement disorder were aggravated in the acacia high-dose +BMS-986299 group. The nuclear condensation phenomenon was intensified and the numbers of Nissl bodies and neurons decreased. The sugar water preference rate, voluntary activity score, and the level of CAT decreased, while the immobility time of forced swimming, the levels of IL-6, IL-β, TNF-α, ROS, MDA, and the expression of HIF-1α, NLRP3, ASC and caspase-1 were increased (P<0.05). Together, these findings demonstrate that acacetin has therapeutic effect on depression rats, and underlying mechanism is related to the inhibition of the HIF-1α/NLRP3 signaling pathway.

This study investigated the role of IL-33 knockdown in attenuating airway inflammation in a mouse asthma model, aiming to identify novel therapeutic targets for asthma. Bone marrow-derived mesenchymal stem cells (BMMSC) were isolated from mice and transduced with IL-33-silencing plasmids (si-IL-33-1^#, si-IL-33-2^#, si-IL-33-3^# and a scrambled control plasmid ) via lentiviral system. The efficiency of IL-33 knockdown was validated by qRT-PCR. An asthma mouse model was established using a combined sensitization and challenge protocol with ovalbumin (OVA). Mice were divided into 5 groups: the control group; the asthma model group; the BMMSC-treated group; the BMMSC+empty vector group and the BMMSC+IL-33 knockdown group. Pathological changes in lung tissues were assessed by H-E staining, Masson's trichrome staining, and immunohistochemistry (IHC). Serum levels of IL-33, IL-1β, and IL-6 were quantified using ELISA. The result showed that primary BMMSC exhibited adherent growth with spindle or polygonal morphology. Flow cytometry was used to measure the expressions of surface markers: CD90 positivity rate was 99.30%; CD29 positivity rate was 100.00%; while hematopoietic lineage markers CD45 and CD34 showed minimal positivity (0.34% and 1.24%, respectively). qRT-PCR demonstrated that the si-IL-33-3^# plasmid effectively silenced IL-33 expression in BMMSC, confirming that it was an optimal IL-33 knockdown construct. Histopathological analysis revealed the BMMSC+IL-33 knockdown group displayed minimal inflammatory cell infiltration in lung tissue, with intact bronchial epithelium. IHC staining showed that BMMSC+IL-33 knockdown group exhibited significantly reduced α-smooth muscle actin (α-SMA) and vascular endothelial growth factor B (VEGFB) relative expressions compared to those in the asthma model group and the BMMSC-treated group (P<0.05). ELISA analysis further confirmed the markedly decreased serum levels of IL-1β, IL-6, and IL-33 in the BMMSC+IL-33 knockdown group compared to either the model or the BMMSC-treated groups (P<0.05). Altogether, IL-33 knockdown attenuates inflammatory response in the asthma mouse model and thereby prevents the lung lesions associated with asthma. 

To investigate the effect of kaempferol on nasal mucosal injury in allergic rhinitis (AR) rats via the TLR4/NF-κB/TNF-α signaling pathway, rat AR model was established. Subjects were randomly divided into the model group, the low-dose (50 mg/kg) kaempferol group, the high-dose (100 mg/kg) kaempferol group, and high-dose (100 mg/kg) kaempferol+LPS (0.4 mg/kg) group as well as the control group. Rhinitis symptoms in rats were observed and scored after drug intervention. H-E staining was applied to observe the pathological changes in rat nasal mucosal tissues. ELISA was used to detect the levels of IL-6, TNF-α in rat nasal lavage fluid (NLF) and of serum IgE. Western blotting was used to detect the protein expressions of TLR4, NF-κB, and TNF-α in nasal mucosal tissues. The results showed that the rhinitis symptom score of the model group was significantly higher than that of the control group (P＜0.05). In addition, pathological changes in the nasal mucosal tissue of the model group were obvious. The levels of IL-6, TNF-α in NLF, IgE in serum, and the protein expressions of TLR4, NF-κB, and TNF-α in nasal mucosal tissues were also significantly increased (all with P＜0.05). Compared to the model group, the medicated rats showed decreased rhinitis symptom scores (both P＜0.05) and alleviated pathological injury. The levels of IL-6, TNF-α in NLF, IgE in serum, and the protein expressions of TLR4, NF-κB, and TNF-α in nasal mucosal tissues were also all decreased. These observations were particularly evident in the high-dose kaempferol group (all with P＜0.05). In contrast, LPS, the TLR4 activator, reversed the effects of kaempferol on AR rats (all with P＜0.05). In summary, kaempferol inhibits the TLR4/NF-κB/TNF-α signaling pathway and alleviates the pathological injury and inflammatory responses of nasal mucosa in AR rats, which in turn improves the disease symptoms.

To explore the neuroprotective effect of Huanglian Jiedu Tang on Parkinson's disease (PD) cell model by the NOD-like receptor family pyrin domain-containing protein 3 (NLRP3) inflammasome pathway, a PD cell model was established by adding 6-hydroxydopamine (6-OHDA) to human neuroblastoma cells (SH-SY5Y). The serum of rats gavaged with Huanglian Jiedu Tang was obtained and cells were treated with different concentrations of drug drug-containing serum or blank serum. The concentration of drug-containing serum was screened using the CCK-8 method. ELISA was used to detect the content of lactate dehydrogenase (LDH) in the cell supernatant. Flow cytometry was used to detect cell apoptosis and the expression of reactive oxygen species (ROS). β-galactosidase staining was used to detect cellular aging. Immunofluorescence assay was used to detect the expression of α-synuclein (α-Syn) and phosphorylated α-Synuclein (p-α-Syn). Western blotting was used to detect the protein expressions of NLRP3, caspase-1 and its precursors pro-Caspase-1, α-Syn, IL-1β and its precursor pro-IL-1β, and IL-18 in each group. Compared with the model group, the low, medium, and high dose Huanglian Jiedu Tang groups showed reduction of LDH content, cell apoptosis rate, and the β-galactosidase increase caused by PD. In contrast, the treatments inhibited the protein expressions of NLRP3, caspase-1, α-Syn, p-α-Syn, IL-1β and IL-18. In summary, Huanglian Jiedu Tang inhibits the inflammation damage of SH-SY5Y cells in PD model induced by 6-OHDA, possibly by regulating the NLRP3 inflammasome pathway, which provides a theoretical basis for the treatment of PD with Huanglian Jiedu Tang.

This study aimed to develop the optimal enzymatic combination for single-cell suspension from mouse lung, and to explore the isolation and culture of the type 2 innate lymphoid cells (ILC2) using immunomagnetic bead separation and flow cytometry. 10 mice were divided into 4 groups for different enzymatic isolation methods, including collagenase Ⅳ, Lung Dissociation Kit, Liberase TM, and collagenase P+dispase Ⅱ, respectively. The most optimal enzymatic isolation method was chosen based on the ratio of living, CD45⁺ and c-kit⁺ cells. Immunomagnetic bead enrichment (EasySep Mouse ILC2 Enrichment Kit) and FACS were used to isolate ILC2 and flow cytometry was used to measure ILC2 homogeneity．Cells cultured in vitro were divided into 2 groups by treatment of IL-33 to evaluate its promoting effect on the in vitro culture of ILC2. Western blotting was used to detect the protein levels of GATA binding protein 3 （GATA-3）, IFN-γ, IL-5 and IL-13. The results indicated that the combination of collagenase P+dispase Ⅱ was the most suitable method for cell isolation of mouse lung with higher ratios of target cell and better cost-effectiveness. The percentage of ILC2 in the enriched fraction reached 2.14% compared to less than 1% before enrichment. The expressions of IFN-γ, IL-5, IL-33 with IL-33 stimulation were higher than those of the control（P<0.001). However, GATA-3 expression showed no statistical significance between the 2 groups（P>0.05). In conclusion，collagenase P+Dispase Ⅱ is a feasible enzymatic digestion protocol for mouse lung tissue. The magnetic cell sorting system could be a rapid and efficient method for ILC2 isolation and future cultivation. In addition, IL-33 can enhance the viability of ILC2 in vitro and stimulate the expression of downstream cytokines. 

The study aims to elucidate the mechanism by which IL-36 regulates monocytes in the pathogenesis of acute B-cell lymphoblastic leukemia (B-ALL). Forty-seven B-ALL patients and 20 controls were enrolled. Serum from patients and controls was collected and PBMCs were isolated. Monocytes and CD4⁺ T cells were purified. ELISA was used to measure and compare the serum IL-36α, IL-36β, IL-36γ, and IL-36 receptor antagonist (IL-36RA) levels between the 2 groups. mRNA levels of IL-36 receptors in monocytes were compared using qRT-PCR. Monocytes were stimulated with recombinant IL-36β or IL-36γ and cell proliferation, granzyme A secretion, granzyme B, granzyme H, IFN-γ, IL-1β, and IL-6 were compared before and after stimulation. mRNA levels of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), Fas ligand (FasL), PD-1 and CTLA-4 expressions in monocytes were also compared. IL-36β- and IL-36γ-stimulated monocytes were co-cultured with autologous CD4⁺ T cells and the Th1 and Th17 percentages were compared. The results showed that, compared to those of controls, the IL-36β and IL-36γ levels were significantly lower in B-ALL patients （P<0.001）. There were no statistical differences of serum mRNA levels of IL-36α/IL-36RA or IL-36 receptor between groups (P>0.05). There were also no significant difference in cellular proliferation, IFN-γ secretion, TRAIL/FasL mRNA, or CTLA-4 expression in monocytes upon stimulation with IL-36β or IL-36γ (P>0.05). The IL-1β and IL-6 secretion increased (P<0.05) while PD-1 expression decreased (P<0.001) compared to those of the unstimulated group. Granzyme A, granzyme B, and granzyme H levels were significantly higher only upon IL-36γ stimulation (P<0.05). There was no significant difference in the percentages of Th1 or Th17 (P>0.05) in the co-culture system of IL-36β or IL-36γ stimulated monocytes and CD4⁺ T cells. This study suggests that IL-36β and IL-36γ levels are down-regulated in B-ALL patients, which might impair the killing activity of monocytes and promote B-ALL progression.

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.

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.

Peripheral helper T cells (Tph) have been identified in recent years as a CD4⁺T cell subset. Tph are characterized by the secretion of C-X-C motif chemokine ligand 13 (CXCL13), and the expression of surface molecules including PD-1, inducible costimulatory (ICOS), HLA-DR, etc. By expressing chemokine receptor C-C motif chemokine receptor 2 (CCR2), C-X3-C motif chemokine receptor 1 (CX3CR1), and CCR5, Tph can be recruited to inflammatory sites and promote the formation of tertiary lymphoid structures (TLS). Through interaction with IL-21 and signaling lymphocytic activation molecule family (SLAMF), Tph assist B cell maturation, differentiation, and antibody production. Tph are shown to be involved in the pathogenesis of many autoimmune diseases. Herein, this review discusses the biological characteristics of Tph and their role in different types of autoimmune diseases to further reveal their mechanism of action and clinical value, and provide new ideas for future targeted therapy.

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.

Primary membranous nephropathy (PMN), an autoimmune disease targeting the kidney, is one of the most common causes of nephrotic syndrome in adults. Although clinical treatments have advanced from supportive care to B cell-targeted therapies including rituximab, challenges such as recurrent delays and significant variability in patient prognoses remain pressing issues. This review discusses the pathogenic mechanisms of auto-reactive B cells in PMN, traces the history of autoantigen discoveries, and provides an update on current B cell-targeted treatments, aiming to offer valuable insights for future research.