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关键词： 黄连-黄芩   CpG1826   细胞因子风暴继发性肺损伤   线粒体膜通透性转换孔(mPTP)   焦亡  

摘要： 该研究旨在探究黄连-黄芩对CpG1826诱导小鼠细胞因子风暴继发性肺损伤（CSSLI）的治疗作用，揭示黄连-黄芩主要效应成分黄连碱和汉黄芩素抑制线粒体膜通透性转换孔（mPTP）/核苷酸结合寡聚结构域样受体蛋白3（NLRP3）炎症小体细胞焦亡路径，并减轻CSSLI的潜在分子机制。体内实验通过CpG1826诱导复制CSSLI小鼠模型，结合肺组织湿干比（W/D）评价肺肿胀程度，苏木素-伊红（HE）染色评价小鼠肺损伤程度，透射电镜（TEM）观察肺组织超微结构，酶联免疫吸附测定（ELISA）检测肺泡灌洗液中白细胞介素（IL）-1β、高迁移率族蛋白B1（HMGB1）、IL-18、IL-1α水平，发现黄连-黄芩水煎液较单味药显著降低肺水肿、肺损伤程度和肺泡灌洗液中相关细胞因子浓度，从而改善CSSLI。体外实验通过小鼠肺泡巨噬细胞（MH-S）建立CpG1826诱导的CSSLI模型，通过钙黄素淬灭筛选黄连-黄芩中抑制mPTP通道开启效果最优单体成分，发现黄连碱(5、10、20 μmol·L－1)和汉黄芩素(10、20、40 μmol·L－1)明显抑制mPTP通道的开启，作用效果最优且呈剂量依赖性；且黄连碱与汉黄芩素通过抑制mPTP通道开放，减少线粒体DNA（mtDNA）的释放和活性氧（ROS）累积，有效逆转CpG1826导致的线粒体膜电位（MMP）降低。进一步研究发现，2个单体成分能抑制细胞焦亡过程，下调NLRP3/半胱天冬蛋白酶-1（Caspase-1）/消皮素D（GSDMD）通路相关蛋白表达。综上所述，黄连-黄芩能改善CpG1826诱导的小鼠CSSLI，与其抑制mPTP/NLRP3细胞焦亡路径有关，为该药对临床应用及开发提供了科学依据。

关键词： 禽网状内皮组织增生症病毒   囊膜蛋白gp90   原核表达   杂交瘤细胞   单克隆抗体  

摘要： 为研制禽网状内皮组织增生症病毒（Reticuloendotheliosis virus,REV）囊膜蛋白gp90单克隆抗体，试验构建重组原核表达质粒pCold-gp90，转化至感受态细胞BL21中进行蛋白诱导表达；采用纯化的gp90蛋白免疫BALB/c小鼠，通过细胞融合技术制备杂交瘤细胞，利用间接酶联免疫吸附试验（ELISA）和流式细胞分选技术筛选鉴定阳性杂交瘤细胞，并制备单克隆抗体腹水，对单克隆抗体的亚型、效价进行测定；并初步测试单克隆抗体在Western blot和间接免疫荧光试验（IFA）中检测gp90蛋白和REV中的应用效果。结果显示：REV囊膜蛋白gp90获得成功表达和纯化，获得3株能稳定分泌REV gp90单克隆抗体的阳性杂交瘤细胞株，该单克隆抗体亚型均为IgG1、Kappa，腹水效价均可达1∶64 000；该单克隆抗体不仅可用于Western blot或IFA检测gp90原核表达蛋白和真核表达蛋白，还可用于IFA检测REV的TCID50效价。研究表明，成功制备的REV gp90单克隆抗体为进一步的REV检测方法建立和gp90蛋白功能研究提供了重要工具。

关键词： Nanocrystals  

摘要： Glaucoma is a group of neurodegenerative eye diseases characterized by progressive damage to the optic nerve which is typically asymptomatic until irreversible vision loss has occurred. Early screening is essential for timely treatment to prevent visual impairment. However, existing detection methods struggle to achieve a balance between accuracy, time efficiency, and portability. The lateral flow assay (LFA) is a well-established immunoanalytical tool for point-of-care (POC) analysis. Here, we have developed a unique dual-testing, quantum nanobeads-based fluorescence LFA, allowing for the simultaneous and quantitative detection of two glaucoma biomarkers: tumor necrosis factor-alpha (TNF-alpha) and brain-derived neurotrophic factor (BDNF). By coating quantum dots on the surface of a SiO2 core, the fluorescent intensity of the quantum nanobeads was enhanced enabling an accurate, efficient, and high-throughput bioanalytical performance, with low detection limits of 3.39 pg mL-1 for TNF-alpha and 4.13 pg mL-1 for BDNF. The LFA also demonstrated superior selectivity, reproducibility, and stability to the standard enzyme-linked immunosorbent assay (ELISA). Using a 3D-printed readout box, the analysis of the LFA requires only a readily accessible smartphone and image processing software, making it an ideal POC detection tool. This ultrasensitive, economical, and user-friendly LFA demonstrates significant potential as an alternative for glaucoma screening.

关键词： Autoimmunity   Cytokines   Interleukin   Keratinocytes   NF-kappa B signaling pathway   Psoriasis   STAT signaling pathways   TNF  

摘要： Psoriasis is a chronic autoimmune disease affecting millions worldwide. This condition is characterized by scaly, red patches of skin that can be painful, itchy, and disfiguring. This non-contagious illness forms plaques and accelerates the dermal cell's life cycle. This review provides a comprehensive overview of the current knowledge on psoriasis, covering its definition, prevalence, causes, pathogenesis, clinical features, diagnosis, and treatment options. The psychosocial impact of psoriasis on patients and their coping mechanisms is also explored. Biologic agents, which target specific cytokines involved in psoriasis pathogenesis, have revolutionized psoriasis treatment and have significantly improved patient outcomes. However, effective and safe treatments for moderate to severe psoriasis are still needed. Future research directions include the development of biomarkers for predicting disease severity and treatment response, investigating new therapeutic targets like the microbiome and epigenetics, and leveraging advancements in technology and genomics for deeper insights into psoriasis pathogenesis and treatment. This study summarizes the key aspects of psoriasis, including its epidemiology, pathophysiology, clinical traits, disease burden, and management. However, further research is needed to improve treatment outcomes and enhance the quality of life for patients affected by this complex condition.

关键词： 单克隆抗体技术   课程教学设计   本科教育   职业教育  

摘要： 目的：为精准培育生物技术人才，确保学生毕业后顺利融入产业的快速发展，本文以《单克隆抗体技术》课程为例，深入探讨教学设计，优化教学方法，提升学生的综合素质，以满足产业对人才的需求。方法：教师通过结合企业实际生产案例、设计小组讨论案例的教学方式，对生物技术专业人才培养模式进行改革与实践。结果：新的课程设计独具匠心，形式新颖，对知识的获取大有裨益。结论：本课程创新地将职业教育元素融入本科教学，有效搭建起课堂与实践的衔接桥梁，深入探索了应用型人才培养的创新路径。

关键词： Mycobacterium tuberculosis   phylogeographic lineages   antigens   HLA binding affinity   host-pathogen interactions   co-evolution  

摘要： Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a global health challenge. The human-adapted TB-causing bacteria are distributed into ten lineages with distinct global distributions and clinical outcomes. Mtb lineages 4 (L4) and L6 are good prototypes of these differences, because L4 is globally prevalent, whereas L6 is geographically restricted to West Africa and associated with slower disease progression. Given the fundamental role of T cells for the control of TB, we questioned whether Mtb L4 or L6 antigens and HLA interactions would be disrupted in West African hosts. Here, we selected variable and validated antigens and demonstrate their expression during in vivo Mtb L4 or L6 infections. We then compared the predicted number of IFN-gamma-inducing and HLA high-binding-affinity peptides in Mtb ancestral, L4, or L6 proteins, considering HLA alleles of high or low frequency in West Africa. Our immunoinformatics approach predicts that non-synonymous substitutions of high variance in Mtb L6 strains diminish binding affinities to HLA alleles prevalent in West African populations, suggesting specific adaptations of these strains to their preferred hosts. Future functional studies will advance our knowledge on lineage-specific evolution and inform strategies to enhance TB control in endemic regions.

关键词： Alport Syndrome   cholesterol   chronic kidney disease   lipid droplet   lipid metabolism   lipotoxicity  

摘要： Starting from the old concept of "fatty degeneration of the kidney", numerous studies have investigated lipid metabolism in chronic kidney disease. Today, it is known that collagenopathies, and more generally diseases involving alterations of the glomerular basement membrane, are characterized by intrapodocyte accumulation of lipid droplets, independent of serum cholesterol levels. Consequently, the latest scientific evidence has focused on the link between the kidney and lipid droplets, ubiquitous but unique organelles, at the same time allies and enemies of renal cells. Copyright by Società Italiana di Nefrologia SIN, Rome,Italy.

关键词： Co-overexpression   PYGO2   IL10   Quantitative real-time PCR   Wnt/beta-catenin pathway  

摘要： Background Esophageal squamous cell carcinoma (ESCC), one of the most aggressive carcinomas of the gastrointestinal tract, is the sixth most common cause of cancer-related death. Wnt pathway plays a pivotal role in cell proliferation and differentiation. PYGO2 and IL10 are involved in this pathway. Our aim in this study was to examine the correlation between PYGO2 and IL10 expression in ESCC patients and cell lines. Methods Relative-comparative real time-PCR (RT-qPCR) was used to evaluate the PYGO2 and IL10 mRNA expression profile in 58 non-treated ESCC compared to their margin normal tissues. Expression of PYGO2 was induced in KYSE-30 and YM1 ESCC lines and IL10 expression was analyzed. Results The results revealed the significant overexpression of PYGO2 and IL10 mRNA in 31.0% and 51.7% of ESCCs, respectively. The PYGO2 and IL10 overexpression was significantly correlated to each other (p = 0.007). Concomitant overexpression of the genes was significantly associated to grade of tumor differentiation (p < 0.01), and tumor depth of invasion (p < 0.05). Induced expression of PYGO2 caused a meaningful change in IL10 expression in ESCC cells. Conclusion PYGO2 may regulate IL10 through Wnt/beta-catenin signaling pathway, suggesting a possible oncogenic role for PYGO2/IL10 axis in ESCC tumorigenesis. Considering the involvement of IL10 as an anti-inflammatory cytokine and PYGO2 role in elevated tumor invasion and metastasis, possible functional interaction between these factors may promote a process which induces invasion and malignant phenotype in ESCC.

关键词： 免疫球蛋白G4相关疾病   肠炎   超声检查  

摘要： 患者男，25岁，10个月前无明显诱因下腹部疼痛，排水样便2～3次/天、偶见脓血，排便后腹痛缓解；2周前因左上腹疼痛就诊于当地医院，考虑“急性胰腺炎”；无特殊家族病史。查体：营养不良、贫血面容，体质量指数16.2 kg/m2；腹软，无压痛及反跳痛。

关键词： CD69基因   原核表达   蛋白纯化   多克隆抗体  

摘要： 为了评估原核表达的猪源CD69蛋白的免疫原性及其潜在的应用价值，以pET-32a(+)为原核表达载体，猪源CD69基因为目的基因，构建重组质粒pET-32a-CD69，针对大肠杆菌BL21(DE3)感受态细胞进行诱导表达；通过变性、复性等方法成功纯化蛋白后，将纯化的CD69蛋白与佐剂按1∶1的比例混合，多次免疫雄性BALB/c小鼠制备多克隆抗体，并用免疫印迹试验(Western-blot)与酶联免疫吸附试验(enzyme linked immunosorbent assay，ELISA)进行鉴定。结果表明，重组的CD69以包涵体形式表达，分子质量约为35 kDa；通过ELISA测定鼠源多克隆抗体效价，结果显示效价达到1∶64 000以上；Western-blot结果表明，该多克隆抗体表现出良好的反应性和特异性。激活标志物CD69蛋白的原核表达及多克隆抗体的制备，为今后我国猪产业的发展与猪相关疾病检测提供了帮助。