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.

ObjectiveTo explore the mechanism by which Qifu Lizhong decoction restores intestinal mucosal barrier function in ulcerative colitis （UC） by regulating the myosin light chain kinase （MLCK）/phosphorylated myosin light chain （p-MLC） signaling pathway. MethodsA rat model of UC with spleen-kidney Yang deficiency syndrome and an in vitro intestinal barrier injury model were established. In vivo experiments were conducted to evaluate the body weight， disease activity index （DAI） scores， and changes in colonic pathological morphology in each group of rats. Enzyme-linked immunosorbent assay （ELISA） was used to measure the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and myeloperoxidase （MPO）. Transmission electron microscopy （TEM） was employed to observe the morphology and structure of tight junctions （TJ） in rat colonic epithelium. In vitro experiments were conducted to observe the morphology of F-actin in human colon adenocarcinoma cell line-2（Caco-2） cells by phalloidin staining. Combining both in vivo and in vitro approaches， immunohistochemistry （IHC）， immunofluorescence （IF）， and Western blot were applied to detect the expression and phosphorylation levels of TJ proteins ［zonula occludens-1 （ZO-1）， Occludin， Claudin-1］ as well as key proteins in the MLCK/p-MLC signaling pathway. ResultsIn vivo experiments demonstrated that， compared with the normal group， the model group exhibited significant body weight loss， elevated DAI scores and histopathological scores （P<0.01）， increased colonic TNF-α， IL-1β， and MPO levels （P<0.01）. TJ structures were disrupted， and the expression of ZO-1， Occludin， and Claudin-1 was significantly reduced （P<0.05， P<0.01）. The protein expression levels of myosin light chain （MLC）， MLCK， phosphorylated （p）-MLC， and p-MLCK significantly increased （P<0.01）. Compared with the model group， all Qifu Lizhong decoction dose groups exhibited increased body weight， as well as significantly decreased DAI scores and histopathological scores （P<0.05， P<0.01）. The colonic levels of MPO， TNF-α， and IL-1β were significantly reduced （P<0.05， P<0.01）. In the Qifu Lizhong decoction + inhibitor group， the expression of TJ proteins ZO-1， Occludin， and Claudin-1 significantly increased （P<0.01）， while the protein expression levels of MLC， MLCK， p-MLC， and p-MLCK were significantly reduced （P<0.01）. In vitro experiment results showed that， compared with the normal group， the TNF-α group exhibited disordered cytoskeletal architecture， weakened fluorescence signals of ZO-1， Occludin， and Claudin-1， and significantly elevated protein expression levels of MLC， MLCK， p-MLC， and p-MLCK （P<0.01）. Compared with the TNF-α group， the Qifu Lizhong decoction + inhibitor group restored the ordered arrangement of the F-actin cytoskeleton， normalized the expression and localization of TJ proteins ZO-1， Occludin， and Claudin-1， and significantly decreased the protein expression levels of MLC， MLCK， p-MLC， and p-MLCK （P<0.01）. ConclusionQifu Lizhong decoction may exert its therapeutic effects on UC by inhibiting the MLCK/p-MLC signaling pathway， thereby stabilizing TJ structure and repairing intestinal barrier integrity.

6-Shogaol is an active component of gingerol in zingiber,which can be converted from 6-Gingerol under acidic and heating conditions.Modern research shows that 6-Shogaol has rich pharmacological activities,and it is found that 6-Shogaol has stronger anti-inflammatory,anti-tumor and antioxidant activities than 6-Gingerol.In this article,the preparation technology and pharmacological effects of 6-Shogaol were reviewed,and the extraction and separation methods of 6-Shogaol,as well as the targets and pathways involved in the process of exerting its pharmacological effects,were summarized,which could lay the foundation for the comprehensive development and clinical application of 6-Shogaol.

Objective·To investigate the changes in transcriptome and chromatin accessibility during the differentiation of human embryonic stem cells(hESCs)into neural progenitor cells(NPCs)using in vitro differentiation models and high-throughput multi-omics sequencing technologies.Methods·hESCs were first induced to differentiate into NPCs in vitro using the embryoid body formation method,and cells at both stages were collected.The cell phenotypes were identified by reverse transcription-quantitative real-time PCR(RT-qPCR)and immunofluorescence(IF)staining.Transcriptome sequencing(RNA-seq)was conducted to detect and analyze the differentially expressed genes(DEGs)between hESCs and NPCs.The assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq)was employed to assess chromatin accessibility changes between hESCs and NPCs.Motif enrichment analysis was performed on differentially accessible chromatin regions to discover potential regulatory transcription factors.Finally,an integrated analysis of RNA-seq and ATAC-seq data and the protein-protein interaction(PPI)network were performed to identify key genes and regulatory pathways involved in the early stages of neural differentiation in vitro.Results·Both RT-qPCR and IF results indicated that the expression levels of pluripotency markers(NANOG and POU5F1)were high at the hESC stage but significantly decreased at the NPC stage,while early neural differentiation markers(PAX6,SOX1,and NES)were minimally expressed at the hESC stage but markedly upregulated at the NPC stage.RNA-seq analysis revealed that compared to the hESC stage,there were 5 597 genes upregulated and 3 654 genes downregulated at the NPC stage.Gene function enrichment analysis showed that the upregulated genes at the NPC stage were enriched in the functions related to neural development.ATAC-seq analysis demonstrated a total of 27 491 genomic regions had significant changes in chromatin accessibility during the differentiation from hESC to NPC,with 12 381 regions showing increased accessibility and 15 110 regions showing decreased accessibility.Motif enrichment analysis revealed that transcription factor genes such as DLX1 and LHX2 might play an important role in the differentiation process from hESCs into NPCs.Integrated analysis of RNA-seq and ATAC-seq data revealed that overlapping genes with high expression at the NPC stage were mainly enriched in axon guidance,forebrain development,and neuron migration.After neural differentiation,the expression levels of CTNND2 and LHX2 genes increased,and the chromatin accessibility of related genomic regions also increased.PPI network analysis indentified candidate downstream genes including PRKACA,CDH2,and ERBB4.Conclusion·The in vitro differentiation model of hESCs combined with high-throughput multi-omics sequencing technologies can be used to depict the changes in transcriptome and chromatin accessibility during the differentiation of hESCs into NPCs.In this process,the expression levels of genes related to axon guidance,forebrain development,and neuronal migration pathways increase and related chromatin accessibility is enhanced.

Objective To analyze the effects of nasal endoscopic low temperature plasma ablation combined with nasal dynamical system-assisted resection on inflammatory markers in peripheral blood of sinonasal inverted papilloma(SINP)and their correlations with recurrence.Methods A retrospective analysis was performed on 60 SINP patients admitted to Chengdu First People's Hospital from May 2021 to May 2023,all of whom were treated with endoscopic low temperature plasma ablation and nasal dynamical system-assisted resection;and another 60 healthy persons who underwent physical examination in the same period were enrolled and set as control group.The changes of peripheral blood inflammatory markers before and at various time points after operation were compared between control group and SINP patients.General data of patients were collected by self-made scales.The recurrence was recorded,and its influence factors were analyzed for further exploring and the correlation between recurrence and levels of peripheral blood inflammatory factors.Results Statistically significant differences between control group and SINP group were found in serum tumor necrosis factor-α(TNF-α),C-reactive protein(CRP),interleukin-6(IL-6),interleukin-8(IL-8),eosinophilic cationic protein(ECP)and eosinophilic granulocytes(EOS)before operation,3 days and 1 month after operation(all P<0.001).Univariate analysis identified that TNF-α,CRP,IL-6,IL-8,mitotic counts,atypical hyperplasia and course of disease were associated with postoperative recurrence in SINP patients.Spearman correlation analysis showed that the postoperative recurrence in SINP patients showed no correlation with IL-6 level(P>0.05),but positively correlated with TNF-α,CRP and IL-8 levels(P<0.05).Conclusion Nasal endoscopic low temperature plasma ablation combined with nasal dynamical system-assisted resection is effective for SINP,with a low postoperative recurrence rate.Moreover,peripheral blood inflammatory markers are closely related to the occurrence and development of SINP,which may be involved in the recurrence,and there is a synergistic effect.

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 the changes in transcriptome and chromatin accessibility during the differentiation of human embryonic stem cells(hESCs)into neural progenitor cells(NPCs)using in vitro differentiation models and high-throughput multi-omics sequencing technologies.Methods·hESCs were first induced to differentiate into NPCs in vitro using the embryoid body formation method,and cells at both stages were collected.The cell phenotypes were identified by reverse transcription-quantitative real-time PCR(RT-qPCR)and immunofluorescence(IF)staining.Transcriptome sequencing(RNA-seq)was conducted to detect and analyze the differentially expressed genes(DEGs)between hESCs and NPCs.The assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq)was employed to assess chromatin accessibility changes between hESCs and NPCs.Motif enrichment analysis was performed on differentially accessible chromatin regions to discover potential regulatory transcription factors.Finally,an integrated analysis of RNA-seq and ATAC-seq data and the protein-protein interaction(PPI)network were performed to identify key genes and regulatory pathways involved in the early stages of neural differentiation in vitro.Results·Both RT-qPCR and IF results indicated that the expression levels of pluripotency markers(NANOG and POU5F1)were high at the hESC stage but significantly decreased at the NPC stage,while early neural differentiation markers(PAX6,SOX1,and NES)were minimally expressed at the hESC stage but markedly upregulated at the NPC stage.RNA-seq analysis revealed that compared to the hESC stage,there were 5 597 genes upregulated and 3 654 genes downregulated at the NPC stage.Gene function enrichment analysis showed that the upregulated genes at the NPC stage were enriched in the functions related to neural development.ATAC-seq analysis demonstrated a total of 27 491 genomic regions had significant changes in chromatin accessibility during the differentiation from hESC to NPC,with 12 381 regions showing increased accessibility and 15 110 regions showing decreased accessibility.Motif enrichment analysis revealed that transcription factor genes such as DLX1 and LHX2 might play an important role in the differentiation process from hESCs into NPCs.Integrated analysis of RNA-seq and ATAC-seq data revealed that overlapping genes with high expression at the NPC stage were mainly enriched in axon guidance,forebrain development,and neuron migration.After neural differentiation,the expression levels of CTNND2 and LHX2 genes increased,and the chromatin accessibility of related genomic regions also increased.PPI network analysis indentified candidate downstream genes including PRKACA,CDH2,and ERBB4.Conclusion·The in vitro differentiation model of hESCs combined with high-throughput multi-omics sequencing technologies can be used to depict the changes in transcriptome and chromatin accessibility during the differentiation of hESCs into NPCs.In this process,the expression levels of genes related to axon guidance,forebrain development,and neuronal migration pathways increase and related chromatin accessibility is enhanced.

Objective To compare the efficacy of recombinant human interleukin-11(rhIL-11)and herombopag olamine tablets on chemotherapy-induced thrombocytopenia(CIT)in breast cancer.Methods Eighty-six prospectively selected breast cancer patients with CIT were randomly divided into the control group(administered rhIL-11)and the study group(administered herombopag olamine tablets),with 43 cases in each group.Before treatment and 14 days after treatment,platelet parameters[platelet count(PLT),plateletcrit(PCT),platelet distribution width(PDW)and mean platelet volume(MPV)]and flow cytometry were measured by automated hematology analyzer.Cellular immune indicators(CD3+,CD4+,CD8+and CD4+/CD8+)were detected by flow cytometer.Enzyme-linked immunosorbent assay was used to determine the levels of thrombopoietin(TPO)and interleukin-11(IL-11).The above parameters were compared between the two groups,and the incidence of adverse reactions in the two groups was recorded.Results After treatment,PLT,PCT,CD3+,CD4+,CD4+/CD8+,TPO and IL-11 in both groups were higher than before treatment,and which was higher in the study group than the control group(P＜0.05).After treatment,PDW,MPV and CD8+of both groups were lower than before treatment,and the study group was lower than the control group(P＜0.05).There was no significant difference in the overall incidence of adverse reactions between the two groups(P＞0.05).Conclusion Both rhIL-11 and herombopag olamine tablets are effective in treating breast cancer with CIT.Compared with rhIL-11,herombopag olamine tablets have more advantages in improving platelet parameters,cellular immune indicators,TPO and IL-11,and do not increase the adverse reactions.

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.

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.