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关键词： Multiattribute method   etanercept   biosimilar   TNF receptor 2-Fc fusion protein   differentiation  

摘要： Ensuring the quality of pharmaceutical products, particularly for complex recombinant protein drugs such as TNF receptor 2-Fc fusion proteins (TNFR2-Fc, Etanercept), poses significant public health challenges. These products, including biosimilars and follow-on versions, exhibit intricate glycosylation patterns and heterogeneous post-translational modifications, complicating their analytical assessment. The Chinese market, hosting four different TNFR2-Fc products, presents a unique regulatory challenge for rapid differentiation and quality control. This study developed a novel mass spectrometry-based multiattribute method (MAM) to address this challenge, enabling simultaneous monitoring of multiple quality attributes and effective differentiation among products from various manufacturers. Conventional techniques initially indicated high purity across all products, but these methods provided limited capabilities for differentiation. The improved MAM approach, involving desialylation, partial deglycosylation, and digestion steps, minimizes heterogeneity and simplifies analysis. This method successfully indicates differences in primary amino acid sequences and specific quality attributes, allowing for a clear differentiation among manufacturers. Notably, products from manufacturers A and B, as well as C and D, despite their high similarity, could be differentiated by their O-glycan profiles. Further activity evaluations revealed that the products from manufacturers C and D exhibited lower binding and biological activity, potentially due to differences in primary amino acid sequences or disulfide bond mismatches. Additionally, all products demonstrated similar Fc-effector functions. In conclusion, this study underscores the variability among TNFR2-Fc products in the Chinese market and the necessity for robust regulatory oversight. The MAM method developed herein serves as a rapid, accurate, and technologically advanced platform for quality control with significant impli

关键词： atrial fibrillation   metabolic syndrome   network pharmacology   sodium-glucose transporter 2 inhibitors   tumor necrosis factor-α  

摘要： Background: Sodium-glucose co-transporter 2 (SGLT2) inhibitors have shown promise in reducing atrial fibrillation (AF) risk, but their mechanism of action in metabolic syndrome (MetS)-related AF remains unclear. This study aims to elucidate the potential mechanisms by which SGLT2 inhibitors interfere with AF initiation in MetS through comprehensive bioinformatic analysis. Methods: A network pharmacology bioinformatic approach was employed to predict the molecular targets of SGLT2 inhibitors. Public databases, including SwissTarget, GeneCards, OMIM, and STRING, were utilized to identify and analyze targets associated with MetS-related AF. Protein–protein interaction (PPI) networks were constructed, followed by gene ontology (GO) and KEGG pathway enrichment analyses to elucidate the biological processes and pathways involved. Results: A total of 52 common targets were identified linking SGLT2 inhibitors with MetS-related AF. The analysis highlighted the TNF-α and AGE-RAGE signaling pathways as key mechanisms through which SGLT2 inhibitors may mitigate AF. Our investigation identified p38 and JNK, components of the TNF-α signaling pathway, as primary targets in reducing atrial remodeling and fibrosis. Conclusions: This study provides insights into the potential mechanisms by which SGLT2 inhibitors influence AF pathogenesis in MetS. The findings suggest that targeting the TNF-α signaling pathway may be a promising therapeutic strategy, with further experimental validation needed to confirm these results. Trial Registration: The authors have nothing to report. © 2025 The Author(s). Journal of Arrhythmia published by John Wiley & Sons Australia, Ltd on behalf of Japanese Heart Rhythm Society.

关键词： 艾滋病   梅毒   男男同性恋   淋巴细胞   快速血浆反应素  

摘要： 目的分析长沙地区男男同性恋HIV/AIDS患者梅毒感染现状,并探讨CD4+T淋巴细胞计数与快速血浆反应素效价的关系。方法采用横断面调查方法,选取2023年1月至2023年12月在长沙市第一医院就诊的4125例HIV/AIDS患者门诊资料,其中合并梅毒感染的264例男男同性恋HIV/AIDS患者作为研究组,同期43名健康体检人群作为对照组。应用流式细胞仪检测两组研究对象外周血CD4+T淋巴细胞计数、CD8+T淋巴细胞计数及两者百分比,血细胞分析仪检测白细胞数(WBC)、中性粒细胞(NE)、血红蛋白(Hb)水平,快速血浆反应素纸片试验检测RPR。双变量线性相关分析合并梅毒感染男男同性恋HIV/AIDS患者CD4+T淋巴细胞亚群的变化与快速血浆反应素效价的相关性。结果4125例HIV/AIDS患者中,男男同性恋人群占52.2%(2153/4125),其中264例合并梅毒感染。异性性取向HIV/AIDS患者占47.8%(1972/4125),其中132例合并梅毒感染。男男同性恋人群HIV/AIDS患者合并梅毒感染率12.3%(264/2153),高于异性性取向梅毒感染率6.7%(132/1972)。研究组CD4+T淋巴细胞百分比、CD4+/CD8+低于对照组,CD8+T淋巴细胞百分比高于对照组,差异有统计学意义(t=7.068、4.000、9.221,P<0.05)。研究组WBC水平及CD8+淋巴细胞计数高于对照组,CD4+T淋巴细胞计数少于对照组,差异有统计学意义(t=1.992、2.314、34.883,P<0.05);两组NE、Hb水平差异无统计学意义(t=0.310、2.870,P>0.05)。双变量线性相关分析结果显示,CD4+T淋巴细胞计数与RPR效价呈负相关(r=-0.130,P=0.020)。结论长沙地区男男同性恋HIV/AIDS患者梅毒感染率高,并高于异性性取向人群,考虑其机体内CD4+T淋巴细胞免疫应答反应失衡是导致快速血浆反应素处于较高效价的原因。

关键词： 白细胞介素-1β   肿瘤坏死因子-α   miR-34a   难治性支原体肺炎患儿   预后  

摘要： 目的探究基线血清白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和miR-34a与难治性支原体肺炎患儿预后的相关性。方法选取宿迁市第一人民医院2022年2月至2025年3月收治的136例难治性支原体肺炎患儿为研究对象。患儿出院后进行1个月随访,根据预后情况分为预后不良组(n=38)与预后良好组(n=98),比较两组的临床资料、血清IL-1β、TNF-α与miR-34a水平。以logistic回归分析筛选难治性支原体肺炎患儿预后不良的独立影响因素后,应用受试者工作特性曲线(ROC)分析基线血清IL-1β、TNF-α和miR-34a对难治性支原体肺炎患儿预后不良的预测效能。结果预后不良患儿支气管充气征比例、胸腔积液比例、临床肺部感染评分(CPIS)、血清IL-1β、TNF-α及miR-34a均较对照组升高,差异有统计学意义(P<0.05);logistic分析显示,支气管充气征比例(OR=1.446)、CPIS评分(OR=1.292)、血清IL-1β(OR=1.178)、TNF-α(OR=1.406)及miR-34a(OR=1.680)水平升高均为难治性支原体肺炎患儿预后不良的独立危险因素(P<0.05);ROC曲线显示,IL-1β、TNF-α、miR-34a三者联合预测难治性支原体肺炎患儿预后不良的曲线下面积为0.916,优于单一检测(P<0.05)。结论基线血清IL-1β、TNF-α和miR-34a与难治性支原体肺炎患儿的预后相关,可为临床诊疗提供参考。

关键词： 抗原修复   免疫组织化学   三羟甲基氨基甲烷(Tris)   乙二胺四乙酸二钠(EDTA)  

摘要： 目的 探究一种新型抗原修复液增强石蜡切片免疫组织化学染色效果。方法 分别用3种不同的抗原修复液(即常规使用的柠檬酸修复液、EDTA修复液,及新配方修复液)对30例不同组织的石蜡组织切片进行加热高压抗原修复,然后选择30个在细胞不同表达部位(细胞膜、浆及核)的抗体完成免疫组织化学染色,观察和统计比较用不同抗原修复液的染色效果(阳性染色染色深浅及阳性细胞数量)。结果 使用柠檬酸修复液组染色+为30.0%(9/30)、++为66.6%(20/30)、+++为3.3%(1/30),EDTA修复液组+为10.0%(3/30)、++为83.3%(25/30)、+++为6.66%(2/30),新型修复液组+为3.3%(1/30)、++为60.0%(18/30)、+++为36.6%(11/30)。结论 新型抗原修复液在多种石蜡组织样本中表现出较好的修复效果,可以缩短抗原修复时间,为组织抗原修复提供了一种新的选择,具有实用价值。

关键词： IL-18   inflammasome   innate immunity   intestine   epithelial cell   stem cell  

摘要： IL-18 is a member of the IL-1 family of cytokines, which is highly expressed in intestinal epithelial cells (IECs). Upon barrier breach, IL-18 is matured to its bioactive form as a result of inflammasome activation, released from the cell via Gasdermin D pores, and sensed by IL-18 receptor 1-positive (IL18R1(+)) immune cells to initiate an inflammatory response. In addition to this epithelial-out signaling network, we recently uncovered an epithelial-intrinsic IL-18 signaling pathway in the murine small intestine and identified enterochromaffin cells and revival stem cells (revSC) as IL18R1(+) bearing IEC populations in the recovering crypt. Here, we discuss the mechanism, and physiological relevance, as well as some of the open questions presented by this discovery in relation to intestinal health and disease.

关键词： 膜性肾病   补体   临床缓解  

摘要： 目的了解膜性肾病患者的尿液补体水平及其与临床缓解的关系。方法收集2018年9月至2023年5月四川省医学科学院·四川省人民医院61例诊断为原发性膜性肾病患者的临床资料和肾活检当日的晨尿标本。通过酶联免疫吸附试验测定尿液中补体因子B(complement factor B,CFB)、补体因子D(complement factor D,CFD)、甘露聚糖结合凝集素相关丝氨酸蛋白酶2(MBL associated serine protease 2,MASP2)、补体C9和膜攻击复合物抑制因子CD59的浓度。通过免疫组化测定CD59和C5b-9在肾组织中的表达情况。收集患者的基线及随访资料,根据2012年改善全球肾脏病预后组织指南,将患者分为临床缓解组(n=35)和不缓解组(n=14),比较两组患者间补体水平的差异。结果在膜性肾病患者尿液中检测到高浓度的CFB、CFD、MASP2、补体C9和CD59。治疗6个月过程中,临床缓解组患者的基线尿CD59水平明显高于不缓解组[164.44(129.80,187.57)μg/L比116.13(68.48,171.77)μg/L,P=0.04]。免疫组化染色提示,临床缓解组患者比不缓解组患者的肾组织CD59表达更高、C5b-9沉积更低。结论膜性肾病患者尿液中可以检测到广泛活化的补体,尿CD59水平与膜性肾病患者临床缓解相关,它可能作为监测患者膜性肾病进展和预测患者膜性肾病预后的潜在生物标志物,并为膜性肾病患者的靶向补体治疗提供线索。

关键词： 多房棘球绦虫   Tetraspanin 5   单克隆抗体   包虫病   CAR-T免疫治疗  

摘要： 制备并鉴定针对多房棘球绦虫(Echinococcus multilocularis)Tetraspanin 5(TSP5)蛋白的单克隆抗体,评估其在包虫病诊断与免疫治疗中的应用潜力。通过基因合成构建pET-28a-TSP5原核表达载体,诱导表达并纯化重组TSP5蛋白;以该蛋白免疫Balb/c小鼠,采用杂交瘤技术筛选阳性杂交瘤细胞株,经有限稀释法进行亚克隆筛选;通过间接ELISA检测抗体效价,采用免疫荧光法分析抗体与天然组织中TSP5蛋白的结合特性,并对抗体可变区序列进行测序与分析。结果显示,本试验成功构建pET-28a-TSP5表达载体,表达并纯化出大小约为8.6 kDa的重组TSP5蛋白(对应其膜外结构域)。经细胞融合与亚克隆筛选,获得一株稳定分泌抗TSP5单克隆抗体的杂交瘤细胞株3B7G12。间接ELISA检测显示该抗体效价达1∶243000,免疫荧光检测结果表明该抗体可特异性识别多房棘球蚴组织中原头蚴与囊壁表面的TSP5蛋白。抗体可变区测序结果显示该抗体重链与轻链CDR区序列明确,具备高亲和力结合潜力。本研究成功制备了高效价、高特异性的抗TSP5单克隆抗体,该抗体能够精准结合多房棘球绦虫组织中的TSP5蛋白,为泡型包虫病的分子诊断提供了新型工具,并为后续CAR-T疗法及抗体药物开发提供了重要依据。

关键词： 强直性脊柱炎   TBNK淋巴细胞亚群   细胞因子  

摘要： 目的探讨强直性脊柱炎(AS)患者外周血TBNK淋巴细胞亚群及细胞因子变化特征,为AS临床诊疗提供新的实验室指标。方法选择125例AS患者作为AS组,其中,HLA-B27阳性患者106例,HLA-B27阴性患者19例,随机选取同期127名健康体检者作为对照组,空腹采集静脉血,采用流式细胞仪测定淋巴细胞亚群CD3^(+)T细胞、CD4^(+)T细胞、CD8^(+)T细胞、B细胞等指标和细胞因子,分析强直性脊柱炎患者外周血TBNK淋巴细胞亚群及细胞因子的变化特征。结果AS组CD4^(+)T细胞占比、CD4^(+)/CD8^(+)明显高于对照组(P<0.05),CD8^(+)T细胞、NK细胞、NKT细胞占比则低于对照组(P<0.05);AS患者组淋巴细胞亚群绝对值均低于对照组(P<0.05)。AS患者组血清IL-6和TNF-α水平更高,TNF-α和IL-6水平与CD4^(+)T细胞呈较强的正相关关系,与CD4^(+)T细胞升高相符。HLA-B27阳性与HLA-B27阴性AS患者淋巴细胞亚群及细胞因子检测结果比较差异无统计学意义(P>0.05)。AS患者BASDAI指数与CD4^(+)T细胞占比、CD4^(+)/CD8^(+)、IL-6和TNF-α水平呈较强的正相关关系。结论强直性脊柱炎患者外周血T淋巴细胞亚群CD4^(+)T细胞占比、CD4^(+)/CD8^(+),细胞因子IL-6和TNF-α水平对AS患者的鉴别诊断和治疗监测具有临床意义。

关键词： Complications   Elderly-onset   Inflammatory Bowel Disease   Mortality   Surgery   5-ASA   5-aminosalycilate   aHR   adjusted hazard ratio   AO   adult-onset   CD   Crohn’s disease   CI   confidence interval   CIF   cumulative incidence function   EO   elderly-onset   HMO   health maintenance organization   HR   hazard ratio   IBD   inflammatory bowel disease   ICD   International Classification of Diseases   NPV   negative predictive value   PPV   positive predictive value   PY   person-years   TNF   tumor necrosis factor   UC   ulcerative colitis   VTE   venous thromboembolism  

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