ObjectiveThis paper aims to investigate the efficacy and related actions of Guizhi Fulingwan in intervening in the mice with colitis-associated colon cancer （CAC） based on the immunosuppressive microenvironment associated with myeloid-derived suppressor cells （MDSCs）. MethodsSixty male C57BL/6 mice were randomly assigned to a blank group， a model group， an aspirin group （0.04 g·kg^-1）， and low-， medium-， and high-dose Guizhi Fulingwan groups （4.87， 9.75， and 19.50 g·kg^-1）， with ten mice per group. The CAC mouse model was established via combined induction of azoxymethane （AOM）/dextran sulphate sodium （DSS）. Drug intervention commenced in week five， with continuous intragastric administration for nine weeks. The food intake， body weight， fecal characteristics， and haematochezia were observed and recorded， and disease activity index （DAI） scores were calculated according to scoring criteria. Hematoxylin and eosin （HE） staining was used to observe the histopathological changes in the colon tissues of the mice. Immunohistochemistry was used to determine proliferating cell nuclear antigen-67 （Ki67） expression in the colon tissues， and enzyme-linked immunosorbent assay （ELISA） was used to detect the contents of interleukin-6 （IL-6）， IL-1β， and tumor necrosis factor-α （TNF-α） in the serum of the mice. Flow cytometry was employed to determine the proportion levels of MDSCs， CD4⁺ T cells， and CD8⁺ T cells in the spleen tissues of the mice. The mRNA expressions of MDSC-associated effector molecules， including arginase 1 （Arg1） and inducible nitric oxide synthase （iNOS）， were detected by real-time quantitative polymerase chain reaction （Real-time PCR）. After that， an in vitro co-culture model of MDSCs and CD8⁺ T cells was established， and drug-containing serum of Guizhi Fulingwan was used for intervention. The Flow cytometry was employed to assess the effects of drug-containing serum of Guizhi Fulingwan with different concentrations on the levels of reactive oxygen species （ROS） and iNOS in MDSCs and the proliferation of CD8⁺ T cells. The levels of granzyme B （GZMB） and interferon-γ （IFN-γ） in cell supernatant were detected by ELISA. ResultsCompared with those in the control group， the mice in the model group exhibited significantly reduced body weight， elevated DAI scores， shortened colon length （P<0.01）， increased number of tumors and Ki67 expression （P<0.01）， and significantly elevated contents of IL-6， IL-1β， and TNF-α in the serum （P<0.01）. Significant increases in the number of MDSCs were observed in mouse spleens， alongside marked reductions in the levels of CD4⁺ T and CD8⁺ T cells （P<0.01）. Furthermore， the mRNA expressions of MDSC function-associated effector molecules Arg1 and iNOS were significantly upregulated （P<0.01）. Compared with those in the model group， the mice in the middle-dose Guizhi Fulingwan group exhibited increased body weight and significantly decreased DAI scores （P<0.05， P<0.01）. The mice in the middle- and high-dose Guizhi Fulingwan groups exhibited significantly improved colon shortening， significantly decreased number of tumors and Ki67 expression （P<0.05， P<0.01）， and significantly decreased contents of IL-6， IL-1β， and TNF-α in the serum （P<0.05， P<0.01）. Furthermore， administration of Guizhi Fulingwan markedly reduced MDSC infiltration in the spleen of the mice， with different degrees of increase in the levels of both CD4⁺ T and CD8⁺ T cells （P<0.05， P<0.01）， alongside significant decreases in the mRNA expressions of Arg1 and iNOS （P<0.05， P<0.01）. In vitro cell co-culture shows that administration of drug-containing serum of Guizhi Fulingwan significantly decreases the activity levels of ROS and iNOS in MDSCs and promotes the proliferation of CD8⁺ T cells and the secretion of GZMB and IFN-γ （P<0.05， P<0.01）. ConclusionGuizhi Fulingwan can reduce pro-inflammatory cytokine secretion and inhibit tumor proliferation in the colon tissues of CAC mice. Its potential mechanism may involve reducing MDSC infiltration， enhancing effector T cells， particularly CD8⁺ T cell response， and improving the tumor immunosuppressive microenvironment.

Background: In acute and chronic renal inflammatory diseases, the activation of inflammatory cells is involved in the defect of erythropoietin (EPO) production. Ras guanine nucleotide-releasing protein-4 (RasGRP4) promotes renal inflammatory injury in type 2 diabetes mellitus (T2DM). Our study aimed to investigate the role and mechanism of RasGRP4 in the production of renal EPO in diabetes. Methods: The degree of tissue injury was observed by pathological staining. Inflammatory cell infiltration was analyzed by immunohistochemical staining. Serum EPO levels were detected by enzyme-linked immunosorbent assay, and EPO production and renal interstitial fibrosis were analyzed by immunofluorescence. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the expression of key inflammatory factors and the activation of signaling pathways. In vitro, the interaction between peripheral blood mononuclear cells (PBMCs) and C3H10T1/2 cells was investigated via cell coculture experiments. Results: RasGRP4 decreased the expression of hypoxia-inducible factor 2-alpha (HIF2A) via the ubiquitination–proteasome degradation pathway and promoted myofibroblastic transformation by activating critical inflammatory pathways, consequently reducing the production of EPO in T2DM mice. Conclusion RasGRP4 participates in the production of renal EPO in diabetic mice by affecting the secretion of proinflammatory cytokines in PBMCs, degrading HIF2A, and promoting the myofibroblastic transformation of C3H10T1/2 cells.

Background: In acute and chronic renal inflammatory diseases, the activation of inflammatory cells is involved in the defect of erythropoietin (EPO) production. Ras guanine nucleotide-releasing protein-4 (RasGRP4) promotes renal inflammatory injury in type 2 diabetes mellitus (T2DM). Our study aimed to investigate the role and mechanism of RasGRP4 in the production of renal EPO in diabetes. Methods: The degree of tissue injury was observed by pathological staining. Inflammatory cell infiltration was analyzed by immunohistochemical staining. Serum EPO levels were detected by enzyme-linked immunosorbent assay, and EPO production and renal interstitial fibrosis were analyzed by immunofluorescence. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the expression of key inflammatory factors and the activation of signaling pathways. In vitro, the interaction between peripheral blood mononuclear cells (PBMCs) and C3H10T1/2 cells was investigated via cell coculture experiments. Results: RasGRP4 decreased the expression of hypoxia-inducible factor 2-alpha (HIF2A) via the ubiquitination–proteasome degradation pathway and promoted myofibroblastic transformation by activating critical inflammatory pathways, consequently reducing the production of EPO in T2DM mice. Conclusion RasGRP4 participates in the production of renal EPO in diabetic mice by affecting the secretion of proinflammatory cytokines in PBMCs, degrading HIF2A, and promoting the myofibroblastic transformation of C3H10T1/2 cells.

Background: In acute and chronic renal inflammatory diseases, the activation of inflammatory cells is involved in the defect of erythropoietin (EPO) production. Ras guanine nucleotide-releasing protein-4 (RasGRP4) promotes renal inflammatory injury in type 2 diabetes mellitus (T2DM). Our study aimed to investigate the role and mechanism of RasGRP4 in the production of renal EPO in diabetes. Methods: The degree of tissue injury was observed by pathological staining. Inflammatory cell infiltration was analyzed by immunohistochemical staining. Serum EPO levels were detected by enzyme-linked immunosorbent assay, and EPO production and renal interstitial fibrosis were analyzed by immunofluorescence. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the expression of key inflammatory factors and the activation of signaling pathways. In vitro, the interaction between peripheral blood mononuclear cells (PBMCs) and C3H10T1/2 cells was investigated via cell coculture experiments. Results: RasGRP4 decreased the expression of hypoxia-inducible factor 2-alpha (HIF2A) via the ubiquitination–proteasome degradation pathway and promoted myofibroblastic transformation by activating critical inflammatory pathways, consequently reducing the production of EPO in T2DM mice. Conclusion RasGRP4 participates in the production of renal EPO in diabetic mice by affecting the secretion of proinflammatory cytokines in PBMCs, degrading HIF2A, and promoting the myofibroblastic transformation of C3H10T1/2 cells.

Background: In acute and chronic renal inflammatory diseases, the activation of inflammatory cells is involved in the defect of erythropoietin (EPO) production. Ras guanine nucleotide-releasing protein-4 (RasGRP4) promotes renal inflammatory injury in type 2 diabetes mellitus (T2DM). Our study aimed to investigate the role and mechanism of RasGRP4 in the production of renal EPO in diabetes. Methods: The degree of tissue injury was observed by pathological staining. Inflammatory cell infiltration was analyzed by immunohistochemical staining. Serum EPO levels were detected by enzyme-linked immunosorbent assay, and EPO production and renal interstitial fibrosis were analyzed by immunofluorescence. Quantitative real-time polymerase chain reaction and Western blotting were used to detect the expression of key inflammatory factors and the activation of signaling pathways. In vitro, the interaction between peripheral blood mononuclear cells (PBMCs) and C3H10T1/2 cells was investigated via cell coculture experiments. Results: RasGRP4 decreased the expression of hypoxia-inducible factor 2-alpha (HIF2A) via the ubiquitination–proteasome degradation pathway and promoted myofibroblastic transformation by activating critical inflammatory pathways, consequently reducing the production of EPO in T2DM mice. Conclusion RasGRP4 participates in the production of renal EPO in diabetic mice by affecting the secretion of proinflammatory cytokines in PBMCs, degrading HIF2A, and promoting the myofibroblastic transformation of C3H10T1/2 cells.

Objective To investigate the relationship between the expression levels of lymphocyte activation gene-3(LAG-3)and pen-traxin-3(PTX3)and the disease severity of the patients with allergic rhinitis(AR).Methods A total of 128 AR patients visited the Department of Otolaryngology,Chengdu Integrated TCM and Western Medicine Hospital from January 2021 to June 2023 were retro-spectively selected as the research subjects(AR group).According to the severity of the condition,the patients were further divided into the mild group(n=40),moderate group(n=42),and severe group(n=46).In addition,80 healthy volunteers who underwent physical examinations in our hospital were selected as the control group.The clinical data such as the gender,age,body mass index(BMI),allergens,and disease onset time of all subjects were recorded and analyzed,and all patients were scored using the Score For Allergic Rhinitis(SFAR).The enzyme linked immunosorbent assay(ELISA)was used to detect the expression levels of serum LAG-3 and PTX3 in all subjects.The correlation between the expression levels of serum LAG-3 and PTX3 was analyzed by the Pearson correla-tion analysis.The receiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of serum LAG-3 and PTX3 levels,both individually and in combination,for AR.Results The expression levels of serum LAG-3 in the AR group(468.74±104.32 μg/L)were significantly lower than that in the control group(691.53±184.65 μg/L,t=7.795,P<0.05),while those of ser-um PTX3 in the AR group(24.83±7.54 ng/L)were significantly higher than that in the control group(17.34±5.37 ng/L,t=7.793,P<0.05).The expression levels of serum LAG-3 in the AR patients with positive immunoglobulin(IgE)and SFAR score≥7 were 442.46±92.37 μg/L and 448.27±103.24 μg/L,respectively,which were significantly lower than that in the AR patients with negative IgE(497.61±115.32 μg/L)and SFAR score<7(498.66±112.76 μg/L,P<0.05).While the expression levels of serum PTX3 in the AR patients with positive IgE and SFAR score≥7 were 28.24±8.17 ng/L and 26.43±8.73 ng/L,respectively,which were significantly higher than that in the AR patients with negative IgE(21.08±6.25 ng/L)and SFAR score<7(22.51±6.89 ng/L,P<0.05).Com-pared with the mild group,the expression levels of serum LAG-3 in both the moderate and severe groups were reduced,and that of ser-um LAG-3 in the severe group was significantly lower than that in the moderate group.While the expression levels of serum PTX3 showed an opposite trend,and the levels of serum PTX3 in the severe group were significantly higher than that in the moderate group(P<0.05).The Pearson correlation results showed a significant negative correlation between serum LAG-3 and PTX3 levels in AR pa-tients(r=-0.402,P=0.000).The analysis of the ROC curve showed that the area under the ROC curve(AUCROC),sensitivity,and specificity of the combination of serum LAG-3 and PTX3 in the diagnosis of AR were 0.881,89.80％,and 73.80％,respectively,which were better than that of serum LAG-3 and PTX3 alone.Conclusion The combined detection of serum LAG-3 and PTX3 for the diagnosis of AR has higher clinical application value and may be used to evaluate the severity of the disease.

Objective:To investigate microbial ecology in restored milk (RM) -donor human milk (DHM) supplemented with mother's own milk (MOM)-under varying MOM ratios, incubation temperatures, and durations. Methods:This in vitro controlled study utilized breast milk samples collected from mothers of preterm infants (<37 weeks) admitted to the Neonatal Intensive Care Unit of Chongqing Health Center for Women and Children between December 2024 and March 2025, including five MOM samples and one DHM sample. Each MOM sample was mixed with DHM at 10% (RM-10 group) or 30% (RM-30 group) volume ratios. Samples were incubated at room temperature (23-26 ℃) and 37 ℃ for 1 h and 4 h, followed by collection. Microbial α-diversity (Chao/Shannon indices), β-diversity (principal co-ordinates analysis), and taxonomic composition (phylum/genus) were analyzed via high-throughput sequencing. Statistical analysis included analysis of variance and the Kruskal-Wallis H test. Results:No statistically significant differences in the Chao index or Shannon index were observed between the RM-10 and RM-30 groups across different incubation times and temperatures ( H or F values=7.61 and 93.20, respectively; both P>0.05). At 37 ℃, the microbial composition of the RM-30 group at both 1 h and 4 h showed no significant difference compared to the initial MOM samples ( R=－0.018, P=0.540), with Firmicutes abundance restored to 65%-90% of the initial MOM level. At room temperature, incubation of the RM-30 group partially restored microbial communities (50%-60%), but induced overgrowth of Proteobacteria (e.g., Pseudomonas, Acinetobacter). Incubation of the RM-10 group at 37 ℃ for 1 h and 4 h also showed no significant difference in microbial composition compared to the initial MOM ( R=－0.004, P=0.442). However, at room temperature, Proteobacteria consistently increased in the RM-10 group samples, and significant differences in microbial composition compared to initial MOM were observed at both 1 h and 4 h ( R=0.179, P=0.027). Conclusion:Under the experimental conditions of this study, preliminary evidence suggests that incubating a blend of DHM and 30% MOM at 37 ℃ for 1 h or 4 h may modulate the microbial composition toward a potentially beneficial profile.

Acute lung injury （ALI） is a clinically critical disease with limited treatment options and poor prognosis， with high morbidity and mortality. Pulmonary inflammation caused by trauma， infection， and other factors in vivo and in vitro can damage alveolar epithelial and vascular endothelial barriers， resulting in lung tissue congestion and edema and eventually leading to significant dyspnea and hypoxemia， It can further develop into acute respiratory distress syndrome. Oxidative stress is one of the pathogenesis of ALI. A large number of reactive oxygen species （ROS） can promote the aggregation of inflammatory cells， increase pulmonary capillary permeability， and even directly damage lung tissue. Therefore， regulating oxidative stress becomes one of the effective means to reduce the degree of lung injury. According to the theory of traditional Chinese medicine （TCM）， ALI is divided into the categories of "sudden wheezing" and "dyspnea due to wheezing". TCM treats the causes of dampness， heat， poison， and stasis by syndrome differentiation and treatment， regulates Qi and blood， and balances Yin and Yang to restore the physiological function of the lung. In recent years， a large number of studies have shown that TCM can regulate ROS through multiple targets and mechanisms and play a role in reducing lung inflammation and protecting alveolar epithelial cells and endothelial vessels， in which the nuclear factor E₂ associated factor 2 （Nrf2） antioxidant pathway plays an important role. Based on the generation and clearance of ROS， this article summarized the related mechanisms of TCM monomers， TCM pairs， and TCM compounds in regulating oxidative stress to prevent ALI， so as to provide theoretical reference for the research and development of new TCM for ALI and clinical treatment.

In 2024, significant advancements have been achieved in the field of pediatric rheumatic and immunological diseases, spanning key conditions such as juvenile idiopathic arthritis (JIA), systemic lupus erythematosus (SLE), and autoinflammatory diseases (AIDs). These developments encompass the indepth elucidation of disease pathogenesis, the expanded application of biomarkers, continuous updates to clinical practice guidelines, and the development of novel targeted therapies. These advances have markedly enhanced the precision and effectiveness of clinical diagnosis and treatment while laying a solid foundation for improving long-term patient outcomes. This article highlights the key research breakthroughs and significant progress in pediatric rheumatic and immunological diseases in 2024, aiming to advance clinical understanding, promote early recognition, and enable accurate diagnosis to optimize outcomes for affected children.

Objective:To analyze the clinical characteristics, treatment, response to treatment, prognosis, and the importance of early recognition and treatment of pediatric systemic lupus erythematosus (SLE) complicated with thrombotic microangiopathy (TMA).Methods:A retrospective summary of the clinical data of 5 children diagnosed with SLE complicated by TMA at Beijing Children′s Hospital, Capital Medical University, from November 2024 to January 2025.Results:Among the 5 children (1 boy and 4 girls, male-to-female ratio of 1∶4), the age of onset ranged from 11 years and 9 months to 14 years and 9 months. All cases had acute onset, rapid disease progression, severe illness, and involvement of multiple organs and systems. The disease activity of SLE was moderately to severely active when TMA was diagnosed. During the course of TMA, all 5 children exhibited varying degrees of hemolytic anemia, thrombocytopenia, renal dysfunction, and proteinuria. Elevated sC5b-9 levels were observed in all 5 children, with 3 showing severely reduced ADAMTS13 activity and 2 with elevated ADAMTS13 inhibitors. Two children had elevated sC5b-9, severely reduced ADAMTS13 activity, and elevated ADAMTS13 inhibitors simultaneously. Four children had newly diagnosed SLE with TMA and achieved stable condition within 2-3 weeks after aggressive treatment, including methylprednisolone pulse therapy, immunosuppressive agents, biologics, plasma infusion, plasma exchange, or dialysis. One child, who had been diagnosed with SLE for 8 years and had irregular oral medication for half a year, suddenly developed TMA with refractory hemolytic anemia and severe thrombocytopenia. This child responded poorly to methylprednisolone pulse therapy, cyclosporine, and eculizumab but showed improvement with plasma exchange. However, the condition was prone to relapse when the interval between plasma exchanges was prolonged. The child eventually responded well to low-dose rituximab and was discharged after 6 weeks of combined treatment. Four children were diagnosed with TMA within 1 week of admission and achieved stable condition after 3 months of follow-up, with no anemia or thrombocytopenia, negative proteinuria, normal complement C3 and C4 levels, and an SLE disease activity score of 0. One child, who had been treated with high-dose glucocorticoid, multiple immunosuppressive agents, and biologics at another hospital, was transferred and diagnosed with TMA 2 weeks after admission. The condition gradually stabilized after treatment with methylprednisolone pulse therapy, immunosuppressive agents, biologics, and plasma exchange. However, at 3-month follow-up, the child still had alopecia and proteinuria, with an SLE disease activity score of 6, indicating mild disease activity.Conclusion:For children with moderate to severe active SLE, especially those with acute onset, rapid disease progression, and prominent manifestations of anemia, thrombocytopenia, renal dysfunction, and poor response to high-dose methylprednisolone pulse therapy, the risk of TMA should be vigilantly monitored. Early diagnosis and aggressive treatment are crucial.