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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。

关键词： 带状疱疹后遗神经痛   神经炎症   胶质细胞   细胞因子   中枢敏化   疼痛环路   Postherpetic Neuralgia   Neuroinflammation   Glial Cells   Cytokines   Central Sensitization   Pain Circuit  

摘要： 带状疱疹后神经痛(PHN)的发病机制复杂，其中神经炎症作为核心驱动因素，通过胶质细胞-神经元交互作用加剧疼痛的慢性化。本文聚焦于神经炎症在PHN中的关键作用，系统阐述病毒诱导的外周炎症如何通过细胞因子级联反应激活中枢胶质细胞，促进脊髓及脑区疼痛通路的敏化与重塑。结合最新研究探讨胶质细胞活化、促炎因子释放及靶向治疗的潜在价值，为PHN的精准干预提供理论依据。

关键词： Endometriosis   N-acetyl cysteine   IFN-gamma   ER stress   Mitochondrial dysfunction   Metabolomics  

摘要： Endometriosis is a chronic inflammatory gynecologic disease characterized by the abnormal implantation of endometrial tissue outside the uterus. The inflammatory microenvironment of endometriosis is dominated by highly migratory endometriotic cells, inflammatory cells, and cytokines. There is no curative treatment other than oral contraceptives, painkillers, and surgery. N-acetyl-L-cysteine (NAC), an anti-inflammatory compound has been identified as a promising agent for endometriosis. However, it is still unclear how NAC interacts with interferon-gamma (IFN-gamma) and common cytokines in the endometriotic microenvironment. This study aimed to investigate the effects of NAC, alone and in combination with IFN-gamma and major cytokines such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) on endometriotic cells. For this purpose, we performed a real time-dependent cell impedance assay, Annexin V/PI and ER tracking by flow cytometry, immunofluorescence, western blotting, and metabolomic assays. Our results offered a new insight into the complex relationship between NAC and IFN-gamma, both of which reduced endometriotic cells' proliferation, induced ER stress and mitochondrial dysfunction. In conclusion, NAC and IFN-gamma, alter the metabolism of endometriotic cells, leading to endoplasmic reticulum stress and mitochondrial dysfunction. These findings suggest that NAC when combined with IFN-gamma, has the potential to generate innovative therapeutic modalities for the treatment of endometriosis.

关键词： N. sativa   thymoquinone   apoptosis   pancreatic cancer   TNF-alpha   NRF2/HO-1  

摘要： Aims This study explores the therapeutic potential of Nigella sativa L. and its key bioactive compound, thymoquinone (TQ).Background Pancreatic cancer presents a significant health challenge due to its aggressiveness and limited treatment options. N. sativa and its component TQ have demonstrated anticancer properties in other cancers, warranting exploration in pancreatic cancer *** To assess the antiproliferative, apoptotic, and anti-invasive effects of N. sativa extracts and TQ on pancreatic cancer cells, with a focus on modulating the NRF2/HO-1 and TNF-alpha signaling *** MIA PaCa-2 and PANC-1 pancreatic cancer cell lines were treated with essential and fixed oils, methanol extracts (from T & uuml;rkiye and Syria), and TQ. Cell viability, apoptosis, and invasiveness were assessed via XTT, Annexin V, and Matrigel assays, respectively. Gene expression and cytokine levels were evaluated using RT-qPCR and ELISA. HPLC was conducted to confirm TQ concentrations in *** The methanol extract of T & uuml;rkiye-originated N. sativa seeds (TM) exhibited the highest cytotoxic effect, reducing cell viability in MIA PaCa-2 and PANC-1 at 0.05 mg/mL, while TQ significantly decreased viability at 20 mu M. TM reduced MIA PaCa-2 and PANC-1 invasiveness (42 +/- 1.23 and 35 +/- 0.73, respectively) and contained a higher concentration of TQ (7.9168 +/- 0.0561%) compared to the Syria-originated extract (SM).Conclusion The findings suggest that TM and TQ exhibit strong anticancer potential by modulating key signaling pathways in pancreatic cancer cells, supporting their potential for further development as therapeutic agents in pancreatic cancer treatment.

关键词： SARS-CoV-2   Omicron variant   Spike protein   T-CoV   HLA  

摘要： BackgroundThe evolution of SARS-CoV-2, particularly through new variants, presents significant global health challenges due to their potential for immune evasion and reduced vaccine effectiveness. This study aims to investigate the impact of mutations in the Spike protein of Omicron EG.5 and XBB.1.16 variants on the binding affinities of viral peptides to common human leukocyte antigen (HLA) class I and II alleles across Taiwanese, British, and Russian populations. Understanding these interactions is crucial for elucidating differences in immune responses and disease severity among diverse *** updated the T-CoV portal to incorporate and analyze EG.5 and XBB.1.16 variants. Binding affinities between mutated Spike protein peptides and HLA class I and II alleles were predicted and compared across the three populations. Statistical analyses, including chi-squared tests, were conducted to assess the significance of binding affinity differences across the three populations and between HLA *** findings revealed that mutations in the Spike protein had a more pronounced effect on HLA class II binding affinities than on HLA class I. While binding affinity profiles for HLA class I were largely consistent across populations, significant population-specific variations were observed for HLA class II alleles. Specifically, the British population exhibited lower proportions of tightly binding mutated peptides compared to the Taiwanese and Russian populations. Furthermore, substantial differences were identified in the binding affinity changes of mutated Spike peptides for HLA class II across Taiwanese, British, and Russian populations, as well as between the Omicron EG.5 and XBB.1.16 variants. Subsequent analyses revealed significant differences in the conservation and evolutionary trajectories of binding affinities between mutated Spike peptides and common HLA class II alleles, both between the EG.5 and XBB.1.16 variants and across the three popula

关键词： Diffuse midline glioma (DMG)   Histone deacetylase (HDAC)   Natural killer (NK) cell   HLA-E  

摘要： BackgroundDiffuse midline glioma (DMG) is an aggressive pediatric brain tumor with limited treatment options. Although natural killer (NK) cell-based immunotherapy is promising, its efficacy remains limited, necessitating strategies to enhance NK cell cytotoxicity. Histone deacetylase (HDAC) inhibition demonstrate potential to enhance NK-mediated killing. However, the combination of HDAC inhibitors and NK cell therapy for DMG remains ***-derived DMG cell lines and orthotopic mouse models were used to evaluate the effects of the class I HDAC inhibitor MS-275 on cytotoxicity. NK cell-mediated lysis was measured using both luciferase and calcein AM-based assays. The downstream signaling pathways affected by MS-275 were investigated via RNA-seq, CUT&Tag assay, RT-qPCR, and chromatin immunoprecipitation with *** on bioinformatic analysis, class I HDACs are identified as therapeutic targets in DMG. The corresponding HDAC inhibitor, MS-275 upregulated NK cell-mediated cytotoxicity pathway through GSEA analysis. Pretreating DMG cells with MS-275 elevated NK cell ligand gene expression and enhanced NK cell-induced lysis. In addition to NK-activating ligands, MS-275 elevated the NK-inhibitory ligand HLA-E, thereby enhancing the efficacy of immunotherapies targeting the NKG2A-HLA-E axis. Mechanistically, MS-275 increased HLA-E expression by promoting STAT3 acetylation at lysine 685. Combining MS-275 with NK cell therapy and blockade of the NKG2A-HLA-E axis extended overall survival in orthotopic mouse *** study is the first to demonstrate that HDAC inhibition enhances NK cell-mediated cytotoxicity in DMG. Combining HDAC inhibition with NK cell therapy represents a promising therapeutic strategy for treating DMG by targeting NKG2A-HLA-E axis.

关键词： 卵巢储备功能减退   炎症细胞因子   Diminished Ovarian Reserve   Inflammatory Cytokines  

摘要： 卵巢储备功能减退(diminished ovarian reserve, DOR)是育龄期女性常见的生殖内分泌疾病，其表现为与年龄、遗传或环境等因素相关的卵子数量和质量下降。长期的慢性炎症会加速身体衰老，从而累及卵巢储备(ovarian reserve, OR)，对女性的生殖健康及心理健康造成负面影响。文章综述了不同炎症因子及相关信号通路与DOR发生发展的研究进展，为寻找DOR的病因及早期临床干预提供新思路。

关键词： 膜性肾病   生物制剂   作用机制   淋巴细胞   补体   临床应用   Membranous Nephropathy   Biologics   Mechanism   Lymphocyte   Complement   Clinical Application  

摘要： 膜性肾病是一种常见的肾小球疾病，其治疗一直是临床关注的焦点。随着对新型足细胞抗原的特异性自身抗体及膜性肾病致病途径的进一步探索，对MN的理解也随之不断发展，除了现在使用的一线生物制剂利妥昔单抗外，新型抗B细胞、抗补体和抗浆细胞等新型生物制剂的应用也在逐渐受到关注。本文阐述了生物制剂在膜性肾病治疗中的研究应用现状，包括其作用机制、疗效等方面，并对其未来的发展前景进行了展望。

关键词： 五氯酚酸钠   单克隆抗体   胶体金免疫层析法   动物性食品   快速检测  

摘要： 为了实现更快速、灵敏、准确检测食品中五氯酚钠的残留，以五氯酚钠的结构为基础，通过保留其苯环上的酚羟基设计合成了半抗原，制备了特异性识别五氯酚钠的单克隆抗体，并建立一种快速检测动物性食品中五氯酚钠的胶体金免疫层析法。通过优化标记体系、抗原包被质量浓度和样品前处理等关键参数，在最优条件下，所制备的抗五氯酚钠单克隆抗体的半数抑制浓度(IC50)为0.87 μg/L，所建立的胶体金免疫层析法对五氯酚钠的标准品检出限、样品检出限分别为0.8、1 μg/kg，具有灵敏、准确、快速等特点。该法可应用于鸡肉、猪肉、桂花鱼、花甲等动物性食品中五氯酚钠的大批量快速筛选。