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关键词： Allograft rejection   HiPSC   Kidney organoid   Kidney transplantation  

摘要： The aim of this study is to establish an in vitro co-culture system to model allograft rejection using kidney organoids system derived from human induced pluripotent stem cells (hiPSCs). We co-cultured kidney organoids derived from wild-type hiPSCs with HLA-mismatched peripheral blood mononuclear cells (PBMCs) from healthy controls (HC) for 24 h. To assess allogeneic rejection modeling, we measured the expression of HLA molecules, (HLA-ABC and HLA-DR), and evaluated cellular damage in the kidney organoids. Additionally, we analyzed the distribution of T cells and their subsets within the co-cultured PBMCs. The immunosuppressive effect of tacrolimus was also evaluated in this co-culture system. Transcriptomic analysis, conducted using RNA sequencing, identified molecules associated with allogeneic rejection. When kidney organoids were co-cultured with alloreactive PBMCs for 24 h, HLA-ABC and HLA-DR expression significantly increased in kidney organoid cells. Additionally, kidney organoids showed reduced cell viability and increased apoptosis compared to syngeneic controls, as assessed by flow cytometry and Annexin V/PI staining. However, treatment with tacrolimus reduced HLA expression in a dose-dependent manner, highlighting the diminished alloimmune responses. Further analysis of PBMC subsets revealed shifts in T helper (TH) and cytotoxic T cell (TC) populations under allogeneic conditions, including increased effector TH and TC cells. Transcriptomic analysis through RNA sequencing identified 256 differentially expressed genes (DEGs), with notable immune-related pathways such as NF-kappa B and TNF signaling involved in allograft rejection. These results provide evidence that a co-culture system with allogeneic kidney organoids and PBMCs can potentially model transplant rejection in vitro.

关键词： 牛结节性皮肤病病毒   山羊痘病毒   A33R蛋白   多克隆抗体   病毒复制  

摘要： 为获得牛结节性皮肤病病毒（LSDV）A33R蛋白的多克隆抗体，并进行免疫学鉴定，本研究利用293F表达系统表达A33R蛋白并验证蛋白的抗原性，将纯化后的重组A33R蛋白作为免疫原3次免疫BALB /c小鼠，获得多克隆抗体。包被不同来源的6种抗原检测多抗的特异性，包被A33R蛋白及灭活羊痘AV41病毒测定其效价，利用中和实验测定多抗的中和效价。此外，利用该抗体测定灭活LSDV及灭活羊痘AV41复制过程中A33R的表达情况。结果表明，本研究成功表达了A33R蛋白并制备了A33R多抗，A33R多抗与LSDV的其它蛋白L1R、A27L、P32不反应，也不与其它常见感染牛或羊的ORFV B2L蛋白、BCoV S蛋白、PPRV N蛋白反应；包被A33R蛋白，多抗效价为1:1 968 300，包被灭活AV41，多抗效价为1:800；进一步测定A33R多抗血清表明血清具有中和效应，且中和效价为 1:64。病毒感染6 h，没有检测到A33R的表达，但是24 h后在LSDV和AV41感染的细胞中均检测到A33R的大量表达。本研究成功的研究结果不仅为进一步研制A33R的单克隆抗体奠定了基础，也为LSDV等痘病毒的复制感染规律的研究和后续疫苗的研发奠定了基础。

摘要： We developed a cell type-specific proteogenomic approach to reveal novel or unannotated open reading frames (nuORFs) using transcriptomics, ribosomal profiling, and proteomic analysis of human pancreatic β-cells using stem cell-derived β-cells and/or cadaveric islets. Our analysis revealed translational regulation of β-cell-specific pathways during differentiation and identified 965 nuORFs, with a majority exhibiting protein support and substantial β-cell specificity. A primate-specific ORF located in the 5' untranslated region of the type 1 diabetes risk gene TYK2 may act as a translational activator. We provide HLA class I immunopeptidomic data from cytokine-stimulated human β-cells and demonstrate their utility in coculture assays with autoreactive T-cell transductants. Taken together, our results define the human β-cell translatome, an important resource to the research field.

关键词： 肥大细胞   MST1   I型超敏反应   综述  

摘要： 超敏反应，是指机体接触抗原并致敏后，再次受到相同抗原刺激时表现出的敏感性增高的免疫反应。其中，由免疫球蛋白E （IgE）和肥大细胞介导的超敏反应称为I型超敏反应。肥大细胞广泛分布于体内各种组织器官的结缔组织和黏膜层，是I型超敏反应中的重要效应细胞。在这一过程中，肥大细胞再次与特异性过敏原蛋白接触时，IgE会与之交联，诱发细胞的激活并脱颗粒，随后产生组胺、细胞因子、趋化因子，以及新合成的花生四烯酸和多种蛋白质来调节I型超敏反应。巨噬细胞刺激因子1（MST1）在细胞生物学过程中扮演着多种重要角色，涉及多种细胞的迁移凋亡、增殖、代谢和细胞骨架调控等领域。本综述将探讨MST1激酶的调控机制以及其对I型超敏反应的影响，以期为过敏性疾病的机制和防控研究提供新的思路。

关键词： CpG   Graves’ disease   IL-10   machine learning model   methylation  

摘要： OBJECTIVE:Epigenetic modifications, particularly deoxyribonucleic acid (DNA) methylation, regulate the expression of immune-mediated factors. This study aimed to investigate the methylation landscape of the interleukin-10 (IL10) gene in Graves' disease (GD). METHODS:This study quantitatively profiled DNA methylation levels within the IL-10 gene using peripheral blood samples from GD patients and healthy controls. Machine learning models were constructed based on CpG methylation features to classify disease status. Furthermore, correlation between specific CpG sites methylation and clinical indicators were analyzed. RESULTS:This study enrolled 60 patients diagnosed with GD and 51 healthy controls. Methylation analysis revealed significantly elevated methylation at multiple IL-10 CpG sites in the GD group compared to the healthy controls ( < 0.01), forming a hypermethylated cluster. Interestingly, the newly-diagnosed GD (NGD) group also showed higher methylation at specific sites compared to the recurrent GD (RGD) group. Correlation analysis showed that methylation at chr1_206947188_R and chr1_206947135_R were positively correlated with FT3 and TRAb levels, indicating that site-specific methylation changes were associated with disease severity and immune activity in GD. CONCLUSIONS:Our findings highlight distinct methylation patterns of the IL-10 gene in GD, with specific CpG sites carrying potential implications for disease diagnosis, stratification, and monitoring.

关键词： 儿童   EBF1-PDGFRB阳性   Ph-like ALL   中枢神经系统白血病   TKIs   达沙替尼   药学监护  

摘要： 报道了1例儿童EBF1-PDGFRB融合基因阳性费城染色体样急性淋巴细胞白血病（Ph-like ALL）合并中枢神经系统白血病（CNSL）的临床病例。患儿在初诊后接受中国儿童白血病协作组（CCLG）-ALL-2018标准化疗方案诱导治疗未获缓解，后续采用含二代酪氨酸激酶抑制剂（TKI）达沙替尼的挽救治疗方案。治疗期间通过治疗药物监测（TDM）及不良反应监测动态调整达沙替尼用药方案，联合方案疗效显著，患儿PDGFRB融合基因及脑脊液微小残留病灶（MRD）持续维持阴性状态。CCLG-ALL-2018化疗方案联合达沙替尼是治疗此类患儿的有效手段，临床药师参与方案优化及实施药学监护对保障用药安全合理至关重要。

关键词： IFN-γ   双调蛋白   肠成纤维细胞   克罗恩病   肠纤维化  

摘要： 目的：克罗恩病（Crohn’s disease,CD）肠纤维化是临床治疗的难点，其分子机制尚未完全阐明。本研究系统探讨干扰素-γ(interferon-gamma,IFN-γ)在CD肠纤维化中的调控作用及机制。方法：提取分离CD患者肠成纤维细胞，体外予以不同的细胞因子（IFN-γ、IL-17A、IL-1β、IL-33、IL-36α、TNF-α）处理，RT-PCR检测人肠成纤维细胞中双调蛋白（amphiregulin,Areg）基因表达水平。20 ng/mL IFN-γ处理人肠成纤维细胞48 h后，进行转录组测序（RNA-seq），结合STRING数据库构建蛋白质互作（protein-protein interaction,PPI）网络。免疫荧光检测α平滑肌肌动蛋白（α-smooth muscle actin,α-SMA）、抗原Kiel 67(antigen Kiel67,Ki67)等肌成纤维细胞标志物，评估IFN-γ对人肠道成纤维细胞活化、增殖的影响。结果：Areg显著促进肠成纤维细胞活化、增殖及胶原合成；IFN-γ可显著抑制肠成纤维细胞Areg表达，并下调α-SMA、COL1A1和COL6A1等纤维化相关基因，同时IFN-γ抑制成纤维细胞增殖与活化能力。RNA-seq分析发现IFN-γ调控的差异基因显著富集于细胞外基质（extracellular matrix,ECM）重构通路，PPI网络鉴定出肉瘤原癌基因（sarcoma proto-oncogene,SRC）为核心节点，提示其可能介导IFN-γ的抗纤维化作用。结论：本研究证实Areg是促CD肠纤维化关键介质，IFN-γ通过转录调控抑制Areg表达，并发现SRC可能是IFN-γ下游的关键效应分子。IFN-γ可能通过抑制SRC/Areg信号轴来发挥其抗纤维化作用。这些发现为开发靶向IFN-γ/SRC/Areg通路的抗纤维化策略提供了理论依据。

关键词： Adaptive immunity   Gastroenterology   Immunology   Inflammatory bowel disease  

摘要： A20, encoded by the TNFAIP3 gene, is a protein linked to Crohn's disease and celiac disease in humans. We now find that mice expressing point mutations in A20's M1-ubiquitin-binding zinc finger 7 (ZF7) motif spontaneously develop proximal enteritis that requires both luminal microbes and T cells. Cellular and transcriptomic profiling reveals expansion of Th17 cells and exuberant expression of IL-17A and IL-22 in intestinal lamina propria of A20ZF7 mice. While deletion of IL-17A from A20ZF7/ZF7 mice exacerbates enteritis, deletion of IL-22 abrogates intestinal epithelial cell hyperproliferation, barrier dysfunction, and alarmin expression. Colonization of adult germ-free mice with microbiota from adult WT specific pathogen-free mice drives duodenal IL-22 expression and duodenitis. A20ZF7/ZF7 Th17 cells autonomously express more RORγt and IL-22 after differentiation in vitro. ATAC sequencing identified an enhancer region upstream of the Il22 gene, and this enhancer demonstrated increased activating histone acetylation coupled with exaggerated Il22 transcription in A20ZF7/ZF7 T cells. Acute inhibition of RORγt normalized histone acetylation at this enhancer. Finally, CRISPR/Cas9-mediated ablation of A20ZF7 in human T cells increases RORγt expression and IL22 transcription. These studies link A20's M1-ubiquitin binding function with RORγt expression, expansion of Th17 cells, and epigenetic activation of IL-22-driven enteritis.

关键词： 脓毒症休克   炎症   免疫   前脓毒素   诊断   相关性  

摘要： 目的 观察血清白细胞介素-6(IL-6)、IL-8、髓系细胞触发受体1(TREM1)、尿激酶型纤溶酶原激活物受体（u PAR）和前脓毒素（presepsin）在脓毒症休克患者中的变化，并分析其诊断价值。方法 前瞻性地选取2023年2月至2025年2月医院收治的150例脓毒症患者作为研究对象，依据病情分为脓毒症（非休克）组（n=44）和脓毒症休克组（n=106）。另择同期各项指标正常的30例健康人作为对照组。比较不同人群血清IL-6、IL-8、TREM1、uPAR和presepsin的水平，采用Spearman相关性分析、logistic回归分析分析血清指标脓毒症病情的相关性，采用受试者工作特征（ROC）评估血清指标诊断脓毒症休克的效果。结果 不同人群血清IL-6、IL-8、TREM1、uPAR及presepsin比较均有差异（P <0.05），呈现脓毒症休克组>脓毒症组>对照组（P <0.05）的趋势。脓毒症休克患者急性生理学和慢性健康状况评价Ⅱ（APACHEⅡ）、序贯器官衰竭估计评分（SOFA）评分均高于脓毒症患者（P <0.05）。Spearman相关性分析结果显示：血清IL-6、IL-8、TREM1、uPAR及presepsin与APACHEⅡ评分、SOFA评分均呈正相关，rs为0.758、0.880、0.837、0.832、0.846、0.487、0.549、0.557、0.626、0.664(P <0.05)。logistic回归模型，结果显示：血清IL-6(OR=1.055)、IL-8(OR=1.054)、TREM1(OR=1.038,95%)、uPAR(OR=1.010)和presepsin(OR=2.103)均与脓毒症休克的发生显著相关（P <0.05）。ROC曲线结果显示，血清IL-6、IL-8、TREM1、uPAR和presepsin诊断脓毒症休克的AUC为0.608、0.724、0.887、0.848、0.885，敏感度为0.432、0.909、0.795、0.909、0.591，特异度为0.880、0.481、0.915、0.736、0.977，多项联合后其诊断脓毒症休克的AUC为0.973，敏感度为0.943，特异度为0.953。且presepsin单独诊断脓毒症休克的AUC高于IL-6、IL-8单独诊断脓毒症休克的AUC(P <0.05)，多项联合诊断脓毒症休克的AUC均高于血清IL-6、IL-8、TREM1、uPAR和presepsin单独诊断脓毒症休克的AUC(P <0.05)。结论 血清IL-6、IL-8、TREM1、u PAR和presepsin在脓毒症休克患者中异常升高，可辅助临床诊断脓毒症休克，5项联合可提升对脓毒症休克的诊断效果。

摘要： Hyperthyroidism, a multifaceted endocrine disorder, is strongly associated with specific human leukocyte antigen (HLA) alleles, which also play a critical role in drug-induced complications such as agranulocytosis. Conventional HLA genotyping methods often face limitations in simultaneously achieving high accuracy, rapidity, and operational simplicity. To address these challenges, we developed a multimodal biosensing platform for the highly sensitive detection of alleles. This platform synergistically integrates three modalities. After molecular recognition, Electrochemiluminescence resonance energy transfer (ECL-RET) was employed using nanomaterials to generate ECL signals, Hybridization chain reaction (HCR) was incorporated to produce fluorescence signals, while nanozyme-mediated colorimetric reactions were coupled to generate visual signals. The combination of these strategies ensures exceptional sensitivity, specificity, and assay stability. Furthermore, spatial separation of the recognition and signal generation domains was implemented to minimize matrix interference from complex biological samples, significantly enhancing clinical applicability. This innovative approach establishes a novel paradigm for the precise analysis of HLA alleles, offering transformative potential for clinical diagnostics and advancing the frontiers of biosensing technologies.