The field of immunometabolism has made significant advances in elucidating the regulation of cellular metabolism on the phenotype and function of immune cells. Succinate is a metabolite of the tricarboxylic acid cycle that plays a critical role as a 'hormone-like' signaling molecule beyond its metabolic function. The macrophages (Mφ) are the hub for immune response, affecting the onset and progression of metabolic diseases associated with inflammation. The polarization of Mφ is shaped by metabolites, which contribute to various disease states and can even lead to metabolic dysfunction in tissues. Succinate has been demonstrated to regulate Mφ phenotype through several mechanisms, promoting or inhibiting inflammatory responses based on environmental factors. Consequently, it is regarded as a key regulator of Mφ function. This review aims to provide an overview of the most recent research on the modulation of Mφ phenotype by succinate-succinate receptor 1 (SUCNR1) axis in inflammatory and fibrotic diseases.

Mast cells are innate immune cells, and matured mast cells are distributed throughout the body, mainly near blood vessels and nerves. Upon stimulation by IgE and other stimuli, degranulation process of mast cells occurs through acascade of signal transduction, releasing inflammatory factors and various cytokines. Mast cells therefore have complicated proinflammatory and immunoregulatory effects. This review focuses on the influence of mast cells on allergic diseases including the promotion of diseases through degranulation, immunoregulation and neuro-immune regulation, highlights the importance of mast cells in autoimmune diseases, discusses their immunoregulatory function in various organs and tissues and their dual effect to autoimmune diseases.

In recent years, the prevalence of obesity has been steadily increasing, yet our understanding of the precise cellular and molecular mechanisms underlying obesity remains limited. The excessive accumulation of adipocytes during the process of obesity leads to a series of low-grade inflammation and has numerous effects on overall physiological function. This review focuses on the significant role of eosinophils in the progression of obesity and related metabolic disturbances, eosinophil generation and development, as well as their recruitment in adipose tissue (AT), and discusses the interaction between the microenvironment of AT and eosinophils. Understanding the detailed characteristics of eosinophils in the AT microenvironment will be beneficial for better exploiting their potential in the treatment of obesity.

To investigate the effect of plumbagin (PL) on neuronal damage in epileptic rats by regulating adenosine-monophosphate-activated protein kinase (AMPK)/cAMP-responsive element-binding protein (CREB)/brain-derived neurotrophic factor (BDNF) signaling pathway, a rat epilepsy model was established by intraperitoneal injection of lithium chloride and pilocarpine. The model rats were randomly divided into model group, low-, medium- and high-dose PL groups (1, 5, and 10 mg/kg, respectively), and inhibitor group (10 mg/kg PL+20 mg/kg AMPK/CREB/BDNF signaling pathway inhibitor Compound C). Rats injected with the equal volume of physiological saline were set as control group. After successful modeling, each group received intraperitoneal injection of corresponding drugs, once a day for 21 consecutive days. The seizure status of rats in each group was evaluated according to the Racine grading standard; Brain waves of rats in each group were observed by electroencephalogram (EEG) and  Morris water maze experiment was applied to evaluate the cognitive and memorial functions. H-E staining was applied to observe the pathological damage of hippocampal tissues and TUNEL staining was applied to observe the apoptosis of rat neural cells. ELISA was applied to detect the levels of IL-6, IL-1β, and TNF-α in the hippocampus of rats in each group. Western blotting was used to detect the expression of proteins regarding the AMPK/CREB/BDNF signaling pathway in hippocampal tissues. Compared to those of the control group, the Racine grading, seizure frequency and duration of rats in the model group were all increased, along with the amplitude, total duration and frequency of the EEG waveform increased. The number of platform crossings and target quadrant activity time were reduced, and the escape latency prolonged. The rats hippocampal tissue structures were seriously damaged, neurons were loosely and badly arranged, with obvious edema and inflammatory cells infiltration. The apoptosis rate of neural cells, IL-6, IL-1β, and TNF-α levels were increased.  The expression levels of p-AMPK/AMPK, p-CREB/CREB, and BDNF proteins were significantly decreased (all with P＜0.05). Compared to those of the model group, the Racine grading, seizure frequency and duration of rats in the low-, medium-, and high-dose PL groups were decreased and the amplitude, total duration and frequency of the EEG waveform was decreased. The number of platform crossings and target quadrant activity time were increased, and the escape latency was shortened. The rats hippocampal structures damage was alleviated to certain extent, the cells were ordered, whose nuclear membrane was relatively clear, with few inflammatory cells infiltration. The apoptosis rate of neural cells, IL-6, IL-1β, and TNF-α levels were decreased. The expression levels of p-AMPK/AMPK, p-CREB/CREB, and BDNF proteins were significantly increased (all with P＜0.05). Compared to those of the high-dose PL group, the changes in the above indicators of rats in the inhibitor group were significantly reversed (all with P＜0.05). In conclusion, PL may activate the AMPK/CREB/BDNF signaling pathway, reduce neuronal apoptosis, inhibit inflammatory responses, and relieve neuronal damage in epileptic rats, thus exerting neuro-protective effects in epileptic rats.

Obesity-induced metabolic disorders, immune dysregulation, and metainflammation promote the pathogenesis and progression of chronic kidney disease (CKD). Apolipoprotein A IV (ApoA-IV) is a lipid-binding protein that regulates gluco-lipid metabolism and exhibits anti-inflammatory activity. However, the impact of ApoA-IV on the immune microenvironment of obesity-related chronic kidney inflammation remains insufficiently explored. This study employed the high-fat diet (HFD) to establish the HFD-induced obesity (DIO) model in both ApoA-IV knockout (ApoA-IV-/-/ApoA-IV KO) and wild type (WT) mice.The results showed that ApoA-IV KO exacerbated insulin resistance and renal lipid accumulation in DIO mice. Further analysis of the single-cell RNA sequencing (scRNA-seq) of kidney tissue and of immune cells revealed that ApoA-IV KO increased oxidative stress in macrophages, aggravated inflammations, impaired macrophage differentiation and polarization of M1- and M2-type phenotypes, reduced antigen processing，presentation and phagosome biological proceedings. Furthermore, ApoA-IV KO weakened the intercellular immune communication. It particularly downregulated the input signaling to macrophages, including pro-inflammatory IFN-II and TNF signaling, and the pro-proliferation and differentiation signaling including CSF and FLT3. Output signaling from macrophages were meanwhile downregulated, including immune-suppressive GALECTIN signaling, pro-resolving ANNEXIN signaling and immune-activating SPP1 signaling. Flow cytometry and bulk RNA sequencing of kidney tissue partially validated the scRNA-seq findings. This study highlights the critical role of ApoA-IV in regulating macrophage differentiation, functioning, oxidative stress, and inflammatory responses in the obese kidney, providing new insights for further exploration of the immune mechanisms underlying chronic kidney inflammation.

In order to evaluate the effect of human mesenchymal stem cells with high expressions of both IL-18 and IL-12 on the in vitro expansion of NK cells from human peripheral blood, IL-18 and IL-12 high expression mesenchymal stem cells were constructed and used as feeding layers in activated culture flasks. PBMCs of healthy humans were collected and cultured in the activation flasks, and a cytokine induced culture protocol was used as the control. Growth curve of NK cell was statistically analyzed. Flow cytometry was used to analyze the proportion of NK cells and K562 cells were used as target cells to examine the in vitro killing ability of NK cells of the stem cell and the control groups. The results showed that the expansion efficiency of NK cells in the absolute ethanol fixed mesenchymal stem cell culture was the highest, and there was no significant difference in the expansion efficiency between the unfixed group and the control group. The purity of NK cells in the fixed group was the highest. The activation culture on day 0 had the highest amplification efficiency and purity, and there was no significant difference in cell purity between activation culture and the 2 activation cultures on day 0 and 5, but the expansion efficiency of NK cells decreased significantly after 2 activation cultures. The killing activity of the stem cell group on K562 tumor cells was significantly better than that of the control group.  Therefore, gene-edited mesenchymal stem cells can improve the in vitro expansion efficiency of NK cells derived from peripheral blood,increase the purity of flow cytometry results, promote the killing activity of NK cell on tumor. The cost of this protocol is lower in addition to being safer and more effective. 

This study aimed to investigate the expression levels of IL-18, IL-18 binding protein a (IL-18BPa), and IL-18 receptor α (IL-18Rα) in plasma CD4⁺ T cells and Th2 of patients with allergic rhinitis and asthma syndrome (ARA) and how allergen regulated their expressions. To this end, blood samples of ARA patients and healthy control subjects were collected, and the expression levels of IL-18, IL-18BPa, and IL-18Rα in CD4⁺ T cells and Th2 were determined by flow cytometry. Plasma levels of  total IL-18 (tIL-18) and total IL-18BPa (tIL-18BPa) were measured by ELISA, and the level of free IL-18 (fIL-18) was calculated. The level of IL-4 was examined by Bioplex system, and the correlations between fIL-18 and IL-4, and the percentage of Th2 were further analyzed. Our data showed that the proportions of IL-18⁺ cells in CD4⁺ T cells and Th2 were increased in ARA patients, and  house dust mite extract (HDME) induced rising IL-18 expression in Th2 of ARA patients. Plasma levels of IL-4, tIL-18, and fIL-18 were increased in ARA patients, and the level of fIL-18 was moderately correlated with levels of IL-4 and the proportion of Th2 in ARA patients. In conclusion, allergen may be involved in the pathogenesis of ARA by inducing elevated expression of IL-18 in Th2.

The study aims to investigate the serum soluble CD39 (sCD39) levels and explore its clinical significance in patients with hepatitis B-related liver diseases. A total of 134 patients with hepatitis B infection were selected for this study, including 46 patients with chronic hepatitis B (CHB), 56 patients with hepatitis B liver cirrhosis (HBLC), 32 patients with hepatocellular carcinoma (HCC), and additional 42 healthy controls (HC). The serum sCD39 levels were detected by ELISA. Additionally, the activities of aspartate transaminase (AST) and alanine transaminase (ALT) were measured by a biochemical analyzer. The expressions of serum AFP, hyaluronic acid (HA), and laminin (LN) were determined by the chemiluminescence method. qRT-PCR was employed to detect the HBV DNA content in serum. Furthermore, the liver fibrosis 4 factor (FIB-4) index, model for end-stage liver disease (MELD) score, AST-to-platelet ratio index (APRI), and modified PAGE-B (mPAGE-B) score were calculated. The differential expressions of serum sCD39 of HBV patients at different stages were measured and Spearman analysis was used to examine the correlation between sCD39 and various clinical indicators. The value of sCD39 in the diagnosis of HCC was evaluated by ROC curve analysis. The results showed that the sCD39 level in the HCC group was the highest and significantly different from that of all other groups (P<0.01). Importantly, the level of sCD39 in patients with liver injury（ALT＞40 U/L）was significantly higher than those without liver injury (ALT≤40 U/L, P<0.001). sCD39 was positively correlated with AST,ALT, AFP, LN, HA, APRI, FIB-4 index (P<0.001), and the MELD score (P=0.016). The AUC of serum sCD39 for the diagnosis of HCC was 0.743. The AUC of AFP combined with sCD39 was 0.894. The study suggests that sCD39 detection may be valuable for prognosis assessment in HBLC patients and for diagnosis of hepatitis B-related HCC. Therefore, sCD39 may serve as a potential diagnostic biomarker for HCC.

The aim of this study was to explore the effect and possible mechanism of total flavonoids of rhizomadrynariae (TFRD) on endometrial damage and cell apoptosis in ovariectomized osteoporotic rats. Postmenopausal osteoporosis (PMOP) rat model was established by ovariectomy. The SD rats were randomly divided into 4 groups: the sham-operation group, the model group, the TFRD treatment group, and the positive drug (estradiol valerate) group. The latter two groups were daily gavaged with 108 mg/(kg·d) TFRD or 0.21 mg/(kg·d) estradiol valerate tablet from the 7th day after operation and treated for 12 weeks. H-E staining was used to observe the pathological morphology of rat femur and uterus tissues. Apoptosis of endometrial cells was evaluated by TUNEL staining. Western blotting was used to detect caspase-3 expression in uterus. ELISA was used to quantify levels of IL-6, TNF-α, and estradiol (E2) in the uterus tissues. Estrogen receptor alpha (ESR1) expression of uterus was detected by immunohistochemistry. Compared to the sham-operation group, the model group showed pathological injury of femur tissues and pathological changes in the uterus tissues. Apoptosis of endometrial cells was increased, along with elevated expressions of caspase-3, IL-6, and TNF-α, while the E2 and ESR1 expressions were decreased (all with P＜0.01). Compared to the model group, the TFRD, and positive drug treatment groups exhibited improved pathological changes in rat femur tissues and uterus. Apoptosis of endometrial cells and the protein levels of caspase-3, IL-6, and TNF-α were significantly decreased, while the expression levels of E2 and ESR1 were increased (all with P＜0.01). Therefore, TFRD can effectively alleviate endometrial injury and apoptosis in PMOP rats, and the mechanism may involve the regulation of E2 and the inhibition of the expression of inflammatory factors IL-6 and TNF-α.

This study aimed to explore the potential mechanism of Nandan bamboo in the treatment of gouty arthritis (GA) by combining network pharmacology and experimental research. Procedures: (1) The pharmacological active ingredients of Nandina domestica Thunb were identified by literature review. The corresponding drug targets were explored by SwissTarget and TargetNet databases. OMIM and GeneCards databases were used to screen for GA disease-related targets. The overlapping targets of the two analyses were imported into the STRING platform for protein-protein interaction (PPI) network analysis to find out the core targets. Cytoscape 3.10.1 software was used to draw the “disease-drug-pathway-target” network to screen for the key active ingredients of Nandina domestica Thunb for the treatment of GA. GO function and KEGG pathway enrichment analysis were performed using online bioinformatics analysis and visualization cloud platform. (2) GA inflammatory cell model was established by stimulation of THP1-derived macrophages using LPS in combination with monosodium urate (MSU). Cells were coincubated with different concentrations of drugs for 24 h. CCK8 assay was used to detect the cell toxicity of Nandina domestica Thunb extract and ELISA was used to detect the IL-1β level in supernatant. RT-qPCR was used to detect the expressions of TLR2/NF-κB signaling pathway related genes and the relative expression of NLRP3 mRNAs. Western blotting was used to detect the NF-κB, NLRP3, and IL-1β protein expressions. The results showed that (1) A total of 309 drug interacting targets, 776 disease-related targets, and 30 common targets were identified. The key bioactive molecules included caffeic acid and biapigenin, and the core disease targets included TNF, PTGS2, RELA (NF-κB p65), SIRT1, and SYK. The key targets were mainly involved in inflammatory signaling pathways such as NF-κB pathway and NOD-like receptor pathway; (2)The CCK8 assay showed that the concentration of Nandina domestica Thunb had no significant effect on cell viability at 37.5, 75 or 150 μg/mL. Compared to those of the control group, the IL-1β level in the cell supernatant, the mRNA expressions of TLR2, myeloid differentiation factor 88 (MYD88), TNF receptor-associated factor 6 (TRAF6), TGF-β-activated activated kinase 1 (TAK1), inhibitor α of κB kinase (IKKα), NF-κB, NLRP3, and IL-1β, and the protein expressions of NF-κB, NLRP3, and IL-1β in the model group were significantly increased (all with P＜0.05). Compared to those of the model group, the level of IL-1β in the cell supernatant, the mRNA expressions of TLR2，MYD88，TRAF6，TAK1，IKKα，NF-κB，NLRP3  and IL-1β, and the protein expressions of NF-κB, NLRP3, and IL-1β in the Nandina domestica Thunb extract treated groups were all significantly decreased（all with P＜0.05）. In conclusion, Nandina domestica Thunb extract can reduce the production and release of inflammatory factors through regulating the TLR2/NF-κB signaling pathway and inhibiting the activation of NLRP3 inflammasome, making it a treatment for GA.

To investigate the protective effect and underlying mechanism of ginkgolide B against trinitrobenzenesulfonic acid（TNBS）-induced ulcerative colitis (UC) in mice, this study used 60 specific pathogen free grade male C57BL/6 mice randomly divided into 6 groups: the control group, the UC model group, the ginkgolide B low-dose group (2.5 mg/kg), the ginkgolide B medium-dose group (5 mg/kg), the ginkgolide B high-dose group (10 mg/kg), and the sulfasalazine group (100 mg/kg). The UC model was established by slowly injecting TNBS solution into the mouse colon through a flexible catheter. The severity of UC was assessed by body weight, colon length, and disease activity index (DAI). Histological changes were observed by H-E staining and the expressions of inflammatory factors were detected by ELISA.The levels of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) were detected by corresponding activity measuring kits. The protein expressions of the TLR4/NF-κB pathway were detected by Western blotting. The results showed that, compared to the model group, mice in both the ginkgolide B medium- and high-dose groups showed less weight loss, longer colon length, lower DAI scores, significantly improved pathological damage of colitis, significantly lower expressions of TNF-α, IL-1β, IL-6, IL-2, and IL-12, significantly higher expression of IL-10, higher levels of SOD and GSH-Px, and lower levels of MDA. In addition, the activation of proteins in the TLR4/NF-κB pathway were inhibited. In summary, ginkgolide B has a significant protective effect on UC, and the mechanism may be related to the inhibition of the activation of TLR4/NF-κB signaling pathway.

In order to investigate the expression of long non-coding RNA (lncRNA) containing antisense RNA1 for 5 PH domains (FGD5-AS1) in hepatocellular carcinoma (HCC) tissues, its effects and mechanism on proliferation, migration, and invasion of HCC cells, HCC tissues and corresponding adjacent tissues of 53 HCC patients were collected.Human HCC cell lines (Hep3B, SK-HEP-1, MHCC97L, MHCC97-H, and Huh-7) and normal human hepatocytes (LO2) were cultured in vitro. RT-qPCR was used to detect the expression levels of lncRNA FGD5-AS1 and miR-512-3p in tissues and cell lines and the correlation between the expression levels of FGD5-AS1 and miR-512-3p in HCC patients' cancer tissues were analyzed by Pearson correlation test.Western blotting was used to detect the expression of Cyclin D1 protein. HCC cell line Huh-7 was cultured in vitro, and was transfected with FGD5-AS1 small-interfering RNA (siRNA) or miR-512-3p mimic, respectively, or co-transfected with FGD5-AS1 siRNA and miR-512-3p inhibitor. The proliferation, migration, and invasion of cells in each group were detected by CCK8 assay, cell cloning experiment and Transwell assay, respectively. The dual-luciferase reporter gene experiment was used to verify the binding between FGD5-AS1 and miR-512-3p or miR-512-3p and Cyclin D1. FGD5-AS1 knockdown (KD) Huh-7 cells were subcutaneously inoculated into nude mice to observe tumor growth. The results showed that compared to those of the adjacent tissues, the protein expressions of FGD5-AS1 and Cyclin D1 were increased in HCC tissues (both P＜0.05), while the expression of miR-512-3p was decreased (P＜0.05). There was a significant negative correlation between FGD5-AS1 and miR-512-3p levels in HCC tissues (r=-0.856, P＜0.01). Compared to those of the LO2 cells, the protein expressions of FGD5-AS1 and Cyclin D1 in human HCC cell lines (Hep3B, SK-HEP-1, MHCC97L, MHCC97-H, and Huh-7) were all increased, while the expression level of miR-512-3p was decreased (all with P＜0.05). FGD5-AS1 KD or over-expression of miR-512-3p reduced the proliferative activity, number of colony formation, migration, and invasion of Huh-7 cells (all with P＜0.05). FGD5-AS1 targeted miR-512-3p, and the expression of miR-512-3p was increased in Huh-7 cells upon FGD5-AS1 KD (P＜0.05). Cyclin D1 was the target gene of miR-512-3p, and the expression of Cyclin D1 protein was decreased in Huh-7 cells overexpressing miR-512-3p (P＜0.05). Down-regulation of miR-512-3p reversed the inhibition of FGD5-AS1 KD on Huh-7 cells proliferation, migration, invasion, and Cyclin D1 expression. Xenografted nude mice result demonstrated that FGD5-AS1 KD inhibited the growth of Huh-7 cells in vivo. In conclusion, lncRNA FGD5-AS1 is highly expressed in HCC tissues and FGD5-AS1 KD inhibits the proliferation, migration, and invasion of Huh-7 cells by targeting the miR-512-3p/Cyclin D1 axis. 

Single-cell RNA sequencing (scRNA-seq) is a novel technigue for high-throughput sequencing analysis of the whole transcriptome to elucidate the gene status of monocells. It is an important research method to investigate the heterogeneity of cells and the underlying mechanisms as well as help to identify novel cell subpopulations. In addition, it provides insight into the molecular underpinnings of disease pathogenesis, evolution, and drug resistance. This technology has quickly developed in recent years and has accumulated a significant amount of research data. This review summarizes the advances of scRNA-seq  and its usage on autoimmune diseases in order to provide a theoretical framework for the application of this technology in immunology and  related fields as well as diagnosis and management of diseases.

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 aim of this study was to prepare a monoclonal antibody targeting the P2 substructural domain of the GⅡ.17 type norovirus (NV) using recombinant protein immunization-hybridoma technique, and to characterize its biological properties. The purified recombinant antigen GⅡ.17-P2 was used to immunize BALB/c female mice, followed by cell fusion of SP2/0 and splenocytes from the immunized mice using the hybridoma technique. A specific antibody positive hybridoma cell line was selected for the production and purification of the monoclonal antibody specific to the substructural domain of the P2 region of GⅡ.17 NV. The biological characteristics of the monoclonal antibody were subsequently evaluated by indirect ELISA, Western blotting, and dot-ELISA assays. Additionally, the sequences of the variable regions of the monoclonal antibody were examined by semi-nested PCR. The results showed that a total of 23 positive hybridoma cell lines were acquired, with a positivity rate of 23.96%. A cell line stably secreting the monoclonal antibody specific to GⅡ.17 type NV P2 domain, named G10, was obtained by subcloning and other techniques. G10 monoclonal antibody was prepared from ascites by antibody purification. The G10 monoclonal antibody had a titer of up to 10-5 μg/mL, a sensitivity of 10-5 μg/mL for antigen detection by means of ELISA, and can recognize the wild-type GⅡ.17 NV. The sequence characteristics of the variable regions of the G10 monoclonal antibody were also elucidated. In brief, monoclonal antibody targeting the P2 substructural domain of GⅡ.17 NV has been successfully developed by the recombinant protein immunization-hybridoma technology, laying the foundation for the development of rapid diagnostic reagents against GⅡ.17 type NV.

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. 

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. 

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.

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.

A special defense mode of neutrophils has been found in recent years, i.e. the neutrophil extracellular trap (NET), which can resist the invasion of pathogens to a certain extent. However, excessive amount of NET will cause pathological inflammation and immune disorders. Studies have confirmed that NET is highly correlated with the imbalance of adaptive immune cells and plays an important role in adaptive immunity. This review briefly summarizes the composition, formation mechanisms, and functional detection of NET, and focuses on the research progress on the relationship between NET and adaptive immune cell imbalance. This review will promote a clear understanding of NET and provide a theoretical basis for revealing the pathogenesis of related diseases, developing disease monitoring and treatment protocols, as well as identifying therapeutic targets.

Acute myeloid leukemia (AML) is a malignant hematological disease characterized by the clonal proliferation of myeloid progenitor cells and high heterogeneity. The development and progression of AML are closely associated with alterations in T cell phenotype and function. These changes can lead to aberrancies in the T cell population, ultimately resulting in immune dysfunction ofthe body. In recent years, researchers have utilized the anti-tumor activity of T cells in the treatment of AML, and it has been observed that the functional recovery of T cells in patients exerts a pronounced inhibitory effect on leukemia cells. At present, T cell immunotherapy for AML includes tumor infiltrating lymphocyte (TIL) therapy, bispecific antibody-recruited T cell therapy, immune checkpoint inhibitor (ICI) therapy, chimeric antigen receptor T-cell (CAR-T) therapy, and tumor-specific receptor gene-transduced T cell (TCR-T) therapy. This review comprehensively summarizes recent domestic and international advancements in T cell immunotherapy research, offering anoverview of the therapeutic efficacy and mechanisms of T cell immunotherapy on AML. The objective is to elucidate new potential strategies for the clinical treatment of AML.

Inflammatory bowel disease (IBD) is a chronic relapsing intestinal inflammatory disease, including ulcerative colitis (UC) and Crohn's disease (CD). Although the etiology of IBD remains unknown, it is well accepted that factors including heredity, intestinal environment, bacterial infection and dysregulated immunity may contribute to the onset. A number of guidelines in China and abroad have pointed out that basal plasmacytosis is an important pathological indicator of IBD diagnosis, and the aggregation value of plasma cells in the middle and lower mucosa is much higher than that of other lymphocytes. It was also observed that plasma cell aggregation disappeared in the intestinal tissue of IBD patients in remission after treatment with biologics. Therefore, it is speculated that the B cell-plasma cell axis in humoral immunity is an important path in the pathophysiology of IBD. However, the underlying mechanism remains unclear. This review summarizes the progress on the roles of humoral immunity of the intestinal mucosa in the pathogenesis of IBD.

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 was to investigate the effect of engeletin (Eng) on the autophagy of neurons induced by oxygen-glucose deprivation/reoxygenation (OGD/R) by the PTEN-induced kinase 1 (PINK1)/Parkin signaling pathway. The mouse hippocampal neuron HT22 cell line was used as the study object to establish an OGD/R-induced cell damaged model. HT22 cells were grouped into the model group (OGD/R group), the low-, medium- and high-dose Eng groups (Eng-L, -M, -H groups, with 1, 5, 10 mmol/L Eng, respectively), the si-PINK1 NC+Eng-H group, the si-PINK1+Eng-H group, and blank group (control group). The cell viability, cell apoptosis, reactive oxygen species (ROS) level, the cell levels of glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and superoxide dismutase (SOD),the changes of mitochondrial membrane potential, cell autophagy level, the expressions of PINK1, Parkin (E3 ubiquitin ligase), microtubule-associated protein 1 light chain 3 (LC3), ubiquitin-binding protein p62, and caspase3 were evaluated using appropriate methods. The results showed that compared to those of the control group, the viability of HT22 cells, the GSH-Px, the SOD levels, the mitochondrial membrane potential, the cell autophagy level, the ratio of LC3-Ⅱ/LC3-Ⅰ, and the protein expressions of PINK1 and Parkin in the OGD/R group were significantly decreased (all with P＜0.05), whereas the apoptosis rate, the caspase3 protein expression, ROS, MDA levels, and p62 protein expression all significantly increased (all with P＜0.05). Compared to those of the OGD/R group, the viability of HT22 cells, the GSH-Px, SOD levels, the mitochondrial membrane potential, the cell autophagy level, the ratio of LC3-Ⅱ/LC3-Ⅰ, and the protein expressions of PINK1 and Parkin in the Eng-L、-M and -H groups were significantly increased (all with P＜0.05) while the apoptosis rate, the caspase3 protein expression, ROS, MDA levels and the p62 protein expression all significantly decreased (all with P＜0.05). Compared to the Eng-H group, si-PINK1+Eng-H treatment reversed the effect of Eng-H on the enhancement of autophagy of HT22 cells, and aggravated cell injury and apoptosis. In conclusion, we have firstly demonstrated that Eng can up-regulate the expression of PINK1/Parkin signaling pathway, and alleviate OGD/R-induced neuronal damage through the mitochondrial autophagy pathway.

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.

Chronic skin wound healing is one of the challenges in the field of repairing. Macrophages play an important modulatoryrole in wound healing and they may polarize to M1 or M2 phenotypes in different microenvironments and upon various stimuli. Dysregulation of the macrophage phenotypes is a vital factor in chronic skin wound healing, and correction of the macrophage phenotypes can promote wound healing. This review summarizesthe origin of skin macrophages, the polarization of macrophages, the mechanism and strategy to regulate macrophages polarization and promote skin chronic wound healing in order to provide evidence for skin chronic wound healing.

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.

The aim of this study is to explore the effect of lncRNA USP2-AS1 on the proliferation, apoptosis and epithelial-mesenchymal transition (EMT) of non-small cell lung cancer (NSCLC) cells through regulating the miR-299-3p/matrix metalloproteinase 2 (MMP2) axis. si-NC, si-USP2-AS1, si-USP2-AS1+inhibitor NC, and si-USP2-AS1+miR-299-3p inhibitor were transfected into A549 cells, respectively, and cells were defined into corresponding groups. Untreated A549 cells were defined as the NC group. qRT-PCR was applied to detect the expressions of lncRNA USP2-AS1 and miR-299-3p in normal human lung epithelial cell line BEAS-2B and A549 cells. CCK-8 assay was used to evaluate the proliferative activity and flow cytometry was used to examine the apoptosis rate of A549 cells.Transwell assay was used to measure the invasion and migration of A549 cells. Western blotting was used to examine the levels of E-cadherin (Ecad), N-cad, vimentin and MMP2 in A549 cells, and dual-luciferase reporting gene assay was used to detectthe interactions of lncRNA USP2-AS1, miR-299-3p and MMP2. Xenograft on nude mice was used to evaluate the growth of tumor cells in vivo. The results showed that compared to those of the BEAS-2B cells, the levels of lncRNA USP2-AS1 and MMP2 in A549 cells were significantly up-regulated (both P＜0.05) while the expression of miR-299-3p was significantly down-regulated (P＜0.05). Compared to those of the NC group and the si-NC group, the proliferative activity, the cell number of migration and invasion, the level of lncRNA USP2-AS1, and the protein levels of N-cad and vimentin in A549 cells in the si-USP2-AS1 group were all significantly decreased (all with P＜0.05) while the apoptosis rate of A549 cells, the expression of miR-299-3p, and the protein level of E-cad were all significantly increased (all with P＜0.05). Compared to those of the NC group and sh-NC group, the graft volume and mass in the sh-USP2-AS1 group were significantly decreased (all with P＜0.05). Inhibition of miR-299-3p expression weakened the effect of lncRNA USP2-AS1 silencing on the inhibition of the proliferation, migration, invasion, and tumor growth of A549 cells, as well as the stimulation of apoptosis. lncRNA USP2-AS1 negatively regulated the miR-299-3p/MMP2 axis. In conclusion, lncRNA USP2-AS1 silencing may down-regulate MMP2 expression through up-regulating miR-299-3p, thereby affecting the proliferation, apoptosis, migration, invasion, and EMT of A549 cells.

To investigate the effect of the different expression levels of  c-Myc  on the differentiation, proliferation, and apoptosis of OT-Ⅰ T cells, OT-Ⅰ T cells were transfected with MSGV c-Myc GFP retrovirus. OT-Ⅰ T cells with high expression of GFP (c-Mychigh group) or low expression of GFP (c-Myclow group) were selected by FACS. qRT-PCR and Western blotting were used to detect the expression of c-Myc in OT-Ⅰ T cells. The expression levels of CD44 and CD62L on the surface of OT-Ⅰ T cells and the expression levels of cytokines IL-2 and Granzyme B (GrB) in OT-Ⅰ T cells were detected by flow cytometry. Western blotting was used to detect the expression levels of proliferating cell nuclear antigen (PCNA) and the proteins that were related to apoptosis in OT-Ⅰ T cells. The proliferation rate of OT-Ⅰ T cells was detected by cell counting. The results showed that high expression of c-Myc promoted T cell differentiation into memory T cells and the proliferation rate of OT-Ⅰ T cells, and enhanced the expression of PCNA protein. Furthermore, higher expression of c-Myc up-regulated the expression of anti-apoptotic protein B cell lymphoma2 (Bcl-2), inhibited the expressions of apoptotic protein Bcl-2-associated X protein (Bax), and the activation and degradation of Caspase-3. In conclusion, the increased expression of c-Myc in OT-Ⅰ T cells plays a role in promoting proliferation and inhibiting apoptosis of OT-I T cells.

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.

The aim of the study was to investigate the effect of alpinetin on intestinal barrier damage in septic rats through regulating the stimulator of interferon gene (STING)/TANK-binding kinase 1 (TBK1)/interferon regulatory factor 3 (IRF3) signaling pathway. Rats were randomly divided into the sepsis group, the alpinetin group, the STING agonist (ADU-S100) group, the alpinetin+ADU-S100 group, and the blank control group, with 18 rats per group. Except the control group, sepsis model was established by cecal ligation and perforation procedure. After successful modeling, corresponding drugs were given once a day for 2 w. The serum level of fluorescein isothiocyanate FITC-glucan in each group was detected. H-E staining was used to evaluate the histopathological injury of small intestine. The expression of zonula occluden 1 (ZO-1) and Occludin in the small intestine was detected by immunohistochemical staining. Mean fluorescence intensity (MFI) of CD86 and CD206 in small intestine tissues was detected by immunofluorescence staining. The levels of TNF-α, IL-6, and IL-10 in the small intestine were detected by ELISA. Western blotting was used to detect the protein levels of STING, p-TBK1, and p-IRF3 in small intestine tissues. The results showed that compared to those of the control group, serum FITC-glucan level, small intestine mucosal Chiu score, CD86 MFI, TNF-α, IL-6 levels and STING, p-TBK1, p-IRF3 protein expressions in small intestinal tissues of sepsis rats were all increased, while the numbers of ZO-1 and Occludin positive cells, CD206 MFI, and IL-10 levels in the small intestine were significantly decreased (all with P<0.001). Compared to those of the sepsis group, serum FITC-glucan level, small intestine mucosal Chiu score, CD86 MFI, TNF-α, IL-6 levels, and STING, p-TBK1, p-IRF3 protein expressions in small intestinal tissues of the alpinetin group were all decreased, while the numbers of ZO-1 and Occludin positive cells in the small intestine, and the levels of CD206 MFI and IL-10 were increased (all with P<0.001). The dynamic trend of the above indexes in ADU-S100 group was opposite to those in the alpinetin group (all with P<0.001). ADU-S100 partially reversed the improvement of intestinal barrier damage in medicated sepsis rats (all with P<0.001). Therefore, the underlying mechanism of alpinetin in improving intestinal barrier damage in sepsis rats is closely associated with its inhibition of the STING/TBK1/IRF3 pathway.

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. 

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 aim of this study is to investigate the effects of long non-coding RNA (lncRNA) plasmacytoma variant translocation 1 (PVT1) on the proliferation, apoptosis, migration, and invasion of cervical cancer cells, and its regulation on miR-484 and Krüppel-like factor 12(KLF12), as well as the IL-6/ STAT3 signaling pathway. qRT-PCR was used to detect the expressions of lncRNA PVT1, miR-484, and the mRNA expression of KLF12 in cervical cancer tissue and cell lines. Double luciferase assay was used to detect the relationship between lncRNA PVT1, miR-484, and KLF12. Upon transfection of Sh-PVT1, Sh-PVT1+antagomir, or Sh-PVT1+antagomir+pcDNA3.1, the proliferation, apoptosis, migration, and invasion of cells were examined. The in vivo tumor growth and metastasis were also measured. Western blotting was used to detect the expression of KLF12, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) in each group of tumor cells and mouse tumor tissue. Immunohistochemistry was used to detect the expression of IL-6, phosphorylated JAK2(p-JAK2), and phosphorylated STAT3 (p-STAT3) in cells and mouse tumor tissue. The results showed that the expressions of lncRNA PVT1 and KLF12 were significantly increased in cervical cancer tissue and cervical cancer cell lines, while the expression of miR-484 was significantly decreased (P＜0.05). Double luciferase assay confirmed the targeted regulation of lncRNA PVT1 on the expression of miR-484 which in turn regulated the expression of KLF12. Inhibiting lncRNA PVT1 significantly inhibited the proliferation, migration, and invasion of cervical cancer cells, induced cell apoptosis, downregulated the expressions of KLF12, Bcl-2, IL-6, p-JAK2, and p-STAT3, and upregulated the expression of Bax (P＜0.05). However, antagomir could partially relieve the suppression of inhibition of lncRNA PVT1 in cervical cancer progression, while pcDNA3.1 partially restored the suppression of inhibition of lncRNA PVT1 in cervical cancer progression. Therefore, this study suggests that in cervical cancer, lncRNA PVT1 upregulates KLF12 by endogenous competition with miR-484 to promote the proliferation, migration, and invasion of cervical cancer cells, inhibit apoptosis, promote the growth and metastasis of tumor cells, and the malignant progression of cervical cancer in vitro, which may be related to the activation of IL-6/STAT3 signaling pathway.

This study aims to explore the role of chemokine C-C motif ligand 20 (CCL20)/C-C chemokine receptor type 6 (CCR6) in unexplained recurrent spontaneous abortion (URSA). Non-pregnant URSA patients (n=22) and early-pregnant URSA patients (n=62) were enrolled as study group, while non-pregnant healthy women (n=30) and early-pregnant healthy women (n=42) were enrolled as control group. CCL20 in peripheral blood were detected by ELISA, and the pregnancy outcomes were followed up. The decidua tissues of URSA (n=36) were collected as the URSA group, and those from normal early pregnant women (n=29) who voluntarily terminated pregnancy during the same period served as the normal group. qRT-PCR was conducted to compare the mRNA expression levels of CCL20 and CCR6 in decidua between the 2 groups. Western blotting was used to compare the protein expressions of CCL20 and CCR6 in decidua between the 2 groups. The expression and localization of CCR6⁺Th17 cells in decidua of the 2 groups were compared by immunofluorescence staining. The results showed that the level of peripheral blood CCL20 in normal early pregnant women was higher than that in URSA early pregnant patients(P<0.05). The mRNA and protein expression levels of CCL20 and CCR6 in decidua of the URSA group were higher than those of the normal group (P<0.05). The number of CCR6⁺Th17 cells in the decidua of URSA patients was higher than that of the normal group. Together, this study suggests that changes in levels of peripheral blood CCL20 have little correlation with pregnancy outcomes. However, at the maternal-fetal interface, CCL20 may chemotactically induce CCR6⁺Th17 cells to migrate and accumulate at the maternal-fetal interface in early pregnancy of URSA patients, causing abortion.

This study aims to investigate the effect of cryptotanshinone (CRY) on the inflammatory response in neonatal rats with necrotizing enterocolitis (NEC) by the JAK2/STAT3 signaling pathway. Ten newborn SD rats were randomly selected as the negative control (NC) group, while the remaining rats were subjected to cold stimulation under hypoxic conditions to establish the NEC model. The successfully modeled rats were randomly divided into the NEC group, the CRY group (7 mg/kg CRY), the JAK2/STAT3 signaling pathway activator coumermycin A1 (C-A1) treatment group (100 μg/kg C-A1), and the CRY+C-A1 group (7 mg/kg CRY+100 μg/kg C-A1), with 10 rats in each group. The NC group and NEC group were given equal amounts of physiological saline, once a day for 2 consecutive weeks. The levels of inflammatory factors and oxidative stress indicators were measured by ELISA. The pathological changes of ileum tissue were detected by H-E staining. Apoptosis of ileal cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. The expressions of JAK2/STAT3 pathway-related proteins were detected by Western blotting. The results showed that compared to those of the NC group, the H-E inflammatory score, levels of IL-1β, IL-6, TNF-α, monocyte chemoattractant protein 1 (MCP-1), malondialdehyde (MDA), apoptosis rate, phosphorylated JAK2 (p-JAK2)/JAK2, phosphorylated STAT3 (p-STAT3)/STAT3 protein levels all significantly increased in the NEC group (P<0.05). On the contrary, the superoxide dismutase (SOD) level significantly decreased (P<0.05). Compared to that of the NEC group, the severity of intestinal tissue lesions in the CRY group decreased, along with the H-E inflammatory score, levels of IL-1β, IL-6, TNF-α, MCP-1, MDA, cell apoptosis rate, p-JAK2/JAK2, p-STAT3/STAT3 protein levels all reduced significantly (P<0.05), while the SOD level significantly increased (P<0.05). In the C-A1 group, the change patterns of above indexes were opposite to those of the CRY group, suggesting that C-A1 eliminated the improvement effect of CRY on NEC rats. Therefore, CRY may improve the inflammatory response in NEC rats by down-regulating the JAK2/STAT3 signaling pathway. 

The aim of this study is to investigate the effect of cinnamaldehyde on intestinal immune function in rats with acute pancreatitis (AP) and its regulation on the Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil protein kinase (ROCK) signaling pathway. Rats were randomly divided into the sham operation group, the model group, the cinnamaldehyde group, the RhoA/ROCK signaling pathway activator lysophosphatidic acid (LPA) group, and the cinnamaldehyde+LPA group, with 15 rats in each group. Except for those in the sham operation group, rats were injected with sodium taurocholate into the pancreatic duct to establish the AP model. Subsequently, corresponding drugs were administered once a day for 2 weeks. ELISA was used to detect the levels of IL-1, TNF-α, and myeloperoxidase (MPO), and H-E staining was used to observe the histomorphology of mesenteric lymph nodes (MLN). The levels of NOD-like receptor family pyrin domain-containing protein 3(NLRP3）and apoptosis-associated speck-like protein（ASC）were detected by immunofluorescence staining. The proportion of Treg and the Th1/Th2 ratio were analyzed by flow cytometry. Western blotting was used to detect the expressions of RhoA/ROCK signaling pathway proteins. The results showed that the structure of lymph nodes in the sham operated group was basically normal, while those of the model group showed an increased number and thickened cortical area. Compared to those of the sham operation group, the levels of IL-1, TNF-α, MPO, the numbers of NLRP3 and ASC positive cells, the ratios of CD4⁺CD25⁺Foxp3⁺, CD4⁺, Treg, Th1, Th1/Th2 ratio, and the protein levels of RhoA and phosphorylated ROCK(p-ROCK)/ROCK in the model group significantly increased (P<0.05), while the level of Th2 significantly decreased (P<0.05).The cinnamaldehyde treatment greatly improved the above indicators, while the LPA treatment had the opposite effect of the cinnamaldehyde treatment and diminished the beneficial effect of cinnamaldehyde on the intestinal immune function in AP rats. In conclusion, cinnamaldehyde may improve the intestinal immune function of AP rats by inhibiting RhoA/ROCK signaling pathway.

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.

As an integral component of the innate immune system,mast cells (MC) are ubiquitously distributed throughout the skin and mucosal tissues. Upon activation by exogenous or endogenous stimuli, these cells rapidly secrete intracellular mediators, including cytokines, proteases, and lipids, thereby playing a crucial role in the defense against bacterial, viral, and parasitic infections, as well as in mediating allergic inflammatory responses. The NOD-like receptor family pyrin domain-containing protein 3 (NLRP3) inflammasome is a critical element of innate immunity, significantly contributing to the body's defense against bacterial and viral infections, and allergen-induced inflammatory responses. Furthermore, it represents a promising therapeutic target for the treatment of various inflammatory diseases.While mast cells and NLRP3 are both integral to the processes underlying allergic inflammatory responses, the mechanisms governing mast cell activation, NLRP3 activation and expression, as well as the regulatory interactions between these components in the context of allergic inflammation remain inadequately elucidated. This review systematically summarizes and discusses the role of mast cell activation and the mediator releasing in modulating NLRP3 activation and expression in the context of allergic inflammatory responses. The objective is to elucidate the mechanisms by which mast cells regulate allergic inflammation, thereby enhancing the comprehensive understanding of the signaling pathways involved in allergic inflammatory responses. This review aims to provide a theoretical foundation for the development of anti-inflammatory treatment strategies and combination therapies.