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关键词： XBP1   Periodontitis   Pyroptosis   Th17/Treg balance   IL-17 Signaling  

摘要： Periodontitis is a long-lasting inflammatory condition that significantly affects people's quality of life. This research focused on examining the function and underlying mechanisms of X-box binding protein 1 (XBP1) in the pathogenesis of periodontitis. In vitro and in vivo models of periodontitis were established using lipopolysaccharide (LPS). The viability and apoptosis of periodontal ligament stem cells (PDLSCs) were assessed using the Cell Counting Kit-8 and flow cytometry assays, respectively. Reverse transcription-quantitative PCR, western blot, and enzyme-linked immunosorbent assays were employed to measure the levels of inflammatory factors and mediators associated with T helper 17 (Th17)/regulatory T cell (Treg) balance, pyroptosis, and the interleukin-17 (IL-17) pathway. Histological and immunohistochemical analyses were conducted to evaluate tissue damage and bone resorption markers. The IL-17 pathway was activated with SR0987 to explore the interactions between XBP1 and IL-17 signaling. XBP1 knockdown reduced apoptosis, pyroptosis, and inflammation, as indicated by lower levels of pro-inflammatory cytokines (TNF-alpha, IL-6, IL-1 beta, IL-18) and pyroptosis-related proteins (ASC, GSDMD-N, NLRP3). XBP1 knockdown alleviated tissue damage, inflammatory cell infiltration, and bone destruction in rat models of periodontitis. XBP1 knockdown notably restored the Th17/Treg imbalance by suppressing Th17 differentiation and promoting Treg differentiation. Mechanistically, XBP1 knockdown inhibited the IL-17 signaling pathway, and IL-17Activator SR0987 significantly reversed the beneficial effects of XBP1 knockdown on periodontitis. XBP1 knockdown alleviated periodontitis by inhibiting the IL-17 signaling pathway, implying that XBP1 could serve as a promising therapeutic target for managing periodontal conditions.

关键词： IBV (传染性支气管炎病毒)   疫苗   重组融合   抗原表位   卵黄抗体  

摘要： 目的：本研究旨在设计并表达融合IBV已证实抗原表位的重组蛋白rMKIBV，作为疫苗以期提供全方位保护，同时探讨经rMKIBV疫苗免疫蛋鸡收获的多克隆抗体卵黄抗体(IgY)在IBV防治中的潜力。方法：通过网站获取的IBV已证实抗原表位序列与GenBank数据库收录的具有代表性的IBV毒株序列进行比对，设计柔性肽连接所有抗原肽。构成的氨基酸序列经过分析处理后，逆翻译并进行密码子优化，随后插入pET-28a(+)克隆载体，导入大肠杆菌进行表达。纯化脱盐、去内毒素处理后的rMKIBV蛋白作为疫苗免疫动物，探究其免疫原性及是否可以刺激蛋鸡产生特异性IgY。结果：检索的IBV抗原表位序列与代表性22株IBV毒株的公开N蛋白和S蛋白序列在相应区域均高度相似。rMKIBV蛋白预测的等电点和分子量分别为10.25和63.39 kDa；二级结构显示无规则卷曲占比最高，表明抗原性较强。成功构建重组质粒pET-28a-mkibv，并在大肠杆菌中表达出高纯度的rMKIBV蛋白。该蛋白能与抗His-tag抗体、N蛋白抗体、S蛋白抗体特异性结合。免疫小鼠后，脾脏指数增大(P<0.05)，血清特异性IgG抗体水平显著升高(P<0.01)，IFN-γ水平升高(P<0.05)，IL-2水平无明显变化。免疫蛋鸡后，成功提取蛋黄中IgY，其特异性IgY抗体水平均显著升高，且IgY抗体滴度在免疫后逐步增大，至50 d左右达到最高，而后缓慢下降，达到稳定水平。结论：本研究成功构建并表达了融合IBV已证实抗原表位的重组蛋白rMKIBV，该蛋白具有较好的免疫原性，能够刺激小鼠和蛋鸡产生特异性抗体。特别是从免疫蛋鸡的蛋黄中获取的IgY，为IBV的防治提供了新的思路，对开发针对IBV的疫苗具有重要的科学意义和应用价值。

关键词： 免疫球蛋白   凝血四项   肾病综合征  

摘要： 探讨在肾病综合征(NS)诊断中,血清免疫球蛋白结合凝血四项的应用价值。方法 选取NS患者62例作为肾病组,30名健康志愿者作为对照组,对入组对象进行血样采集和测定,包括血清免疫球蛋白[免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、免疫球蛋白A(IgA)]、凝血四项[凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、凝血时间(TT)、纤维蛋白原(Fg)]、肾功能指标[肌酐(Cr)、血尿酸(UA)、估算肾小球滤过率(eGFR)]。根据对比结果完成组间对比、相关性和ROC曲线分析。结果 肾病组血清免疫球蛋白三项指标、凝血四项指标与肾功能三项指标测定结果均异于对照组(P<0.05);肾病失代偿组血清免疫球蛋白三项指标、凝血四项指标与肾功能三项指标测定结果均异于代偿组(P<0.05);免疫球蛋白、凝血四项与肾功能指标均表现出相关性(P<0.05);ROC曲线分析,免疫球蛋白联合凝血四项的曲线下面积(AUC)为0.899,敏感度和特异度分别为95.16%、80.65%。结论 免疫球蛋白联合凝血四项在NS诊断中具有较高应用价值。

关键词： COVID-19   代谢组学   细胞因子   犬尿氨酸  

摘要： 目的 分析新型冠状病毒感染（coronavirus disease 2019,COVID-19）患者多种生物标本代谢水平的变化，挖掘COVID-19疾病进展的生物标志物，为构建预测模型和发现潜在治疗靶点提供理论依据。方法 采集2022年12月—2023年1月于北京大学第三医院呼吸与危重症医学科、普外科和重症监护病房住院的45例COVID-19患者的血清、尿液和粪便样本，使用液相色谱质谱联用方法检测标本的代谢物水平。采用差异倍数（fold change,FC）、t检验和Wilcoxon秩和检验筛选差异代谢物。对代谢物与血清细胞因子水平进行相关性分析并计算斯皮尔曼等级相关系数。结果 色氨酸代谢通路的犬尿喹啉酸在重症患者血清中水平上调（FC=2.47,P=0.035），喹啉酸在死亡结局患者血清中水平上调（FC=2.99,P=0.010）。血清犬尿喹啉酸在出现多脏器衰竭（FC=4.74,P=0.013）的患者中代谢水平上调，尿液黄尿酸在出现肾衰竭的患者中代谢水平上调（FC=4.40,P=0.019）。相关性分析结果显示，重症患者血清犬尿喹啉酸与辅助性T细胞（r=-0.75,P<0.001）有较强负相关，死亡结局患者血清喹啉酸和总补体（r=-0.88,P=0.007）有较强负相关。COVID-19患者在血清、尿液和粪便中均表现出犬尿氨酸代谢途径上调的趋势。结论 犬尿氨酸代谢通路在COVID-19疾病进展和免疫调控中具有重要作用，犬尿喹啉酸、喹啉酸等代谢物的水平失调可能与免疫系统激活和不良预后有关。

关键词： Aryl hydrocarbon receptor   Checkpoint Control   Gamma interferon   Immunotoxicology   Long noncoding RNA   Lung cancer   Polycyclic aromatic hydrocarbons  

摘要： Although immune checkpoint inhibitors show great promise, not all patients respond and many do not achieve durable responses. Consequently, further investigations into potentially targetable molecules that regulate immune checkpoints are warranted. Previous studies in several cancers demonstrated that interferons produced by tumor-infiltrating leukocytes regulate immunosuppressive PD-L1, PD-L2 and IDO1 through JAK/STAT signaling. Here, we investigated a novel role for an immunosuppressive environmental chemical receptor, previously implicated in smoking-related cancers, in IFN signaling in human lung adenocarcinoma (LUAD) cells. Deletion of the aryl hydrocarbon receptor (AhR) from A549 LUAD cells significantly decreased baseline JAK2, STAT1, STAT3, IRF1 (a JAK/STAT target), PD-L1, PD-L2, and IDO1 expression. IFNγ and IFNα increased expression of JAK/STAT and immune checkpoint genes and proteins, but these increases were significantly diminished or absent in AhR-knockout cells. The AhR similarly controls IFN-induced, JAK/STAT-driven increases in multiple MHC class I- and class II-related genes. AhR control of type I and type II interferon signaling is mediated through up-regulation of a lncRNA, the IFN-stimulated non-coding RNA 1 (INCR1), and through repression of an RNA-binding protein, heterogeneous nuclear ribonucleoprotein H1 (HNRNPH1), which sequesters JAK/STAT-related and immune checkpoint gene transcripts. The data suggest that the AhR is a key mediator of tumor immunosuppression through regulation of IFN-induced INCR1 and JAK/STAT signaling and, thereby, expression of immune checkpoints. However, that immunosuppression may be tempered by AhR control of MHC expression. Given the multiple roles of JAK/STAT signaling in the immune system, the results also suggest multiple levels on which the AhR may affect tumor immunity.

关键词： Gpr109A   Macrophages   Hepatocellular carcinoma   Immune microenvironment   MerTK   Proteases  

摘要： To delineate Gpr109A's role and mechanisms in modulating the immune microenvironment of hepatocellular carcinoma. Employing Gpr109A-knockout mice and in vitro co-cultures of hepatocellular carcinoma cells with macrophages, this study utilized a suite of techniques, including lentiviral vectors for stable cell line establishment, Western blotting, cell scratch, CCK-8, transwell assays, flow cytometry, immunohistochemistry and phagocytosis assay to assess various cellular behaviors and interactions. Gpr109A deletion markedly reduced the oncogenic potential of H22 cells, both in vivo and when co-cultured with knockout macrophages, impairing their growth, invasion, and migration. In Gpr109A-knockout macrophages, an upregulation of MerTK and a reduction in immunosuppressive cytokine release were observed, indicating a shift towards an M2c macrophage phenotype. This shift is linked to Gpr109A's role in promoting protease overexpression and inhibiting SHP2 phosphorylation, crucial for enhancing cancer cell proliferation and invasiveness. Gpr109A significantly influences macrophage polarization to the M2c type, augmenting hepatocellular carcinoma cell aggressiveness.

关键词： 牛冠状病毒   多克隆抗体   双抗体夹心ELISA   鉴定  

摘要： 不断发现的新型冠状病毒对人类的危害极其严重，牛冠状病毒（BCoV）具有致人畜共患病的潜力，而BCoV分离鉴定及病原检测均缺乏有效的方法。本研究制备与BCoV发生特异性反应的家兔多克隆抗体，为BCoV的检测和鉴定奠定技术基础。本研究将BCoV灭活后免疫家兔制备针对病毒的多克隆抗体。经3次免疫后检测血清中和效价，将纯化后的多克隆抗体用于间接免疫荧光（IFA）、免疫印迹（Western-blot）和夹心ELISA试验。结果显示，制备的家兔多克隆抗体与BCoV反应，血清中和效价可达到1∶28，与cpCoV反应，可达1∶29。经IFA鉴定，该多克隆抗体能够与不同毒株的BCoV和cpCoV发生特异性反应；Western-blot分析发现在约50 kDa处出现特异性条带；将家兔多克隆抗体作为夹心ELISA的包被抗体，只与BCoV和cpCoV发生特异性反应，而不与其他病毒发生交叉反应。该多克隆抗体与不同毒株的BCoV和cpCoV特异性结合，可用于分离鉴定BCoV和cpCoV的IFA和Western-blot分析，制备的多克隆抗体还可作为包被抗体用于检测BCoV与cpCoV的双抗夹心ELISA。本研究为BCoV与cpCoV的病原检测与流行病学调查奠定了基础。

摘要： The cytogenetic risk category retains a pivotal role in the prediction of prognosis in acute myeloid leukemia (AML) patients undergoing hematopoietic stem cell transplantation (HSCT), however, its impact on secondary AML (sAML) is less established. We assessed whether the ELN 2022 cytogenetic risk score predicts outcomes in sAML patients in remission undergoing HSCT from HLA-matched donors performed between 2010 and 2022. Among 1119 patients, 829 had intermediate and 284 had adverse cytogenetics (6 with favorable risk were excluded). Engraftment rates was 72.4% vs. 99.5%. Acute graft-versus-host disease (GVHD) incidence did not differ, but 2-years all grades and extensive chronic GVHD were higher in the intermediate vs. adverse cytogenetics risk groups, hazard ratio (HR) = 0.72;p = 0.034 and HR = 0.58;p = 0.027, respectively. Two-year non-relapse mortality (NRM) was similar. All other HSCT outcomes were inferior in the adverse risk vs. intermediate-risk patients: The HR for 2-year relapse incidence (RI) was 2.48 (95% CI 1.95-3.15, p < 0.001). The HRs for 2-year leukemia-free survival (LFS), overall survival (OS), and GVHD-free/relapse-free survival (GRFS) were 1.62 (95% CI 1.34-1.95, p < 0.001), 1.59 (95% CI 1.3-1.93, p < 0.001) and 1.38 (95% CI 1.15-1.65, p < 0.001), respectively. We conclude that cytogenetic risk score predicts HSCT outcomes in sAML patients.

关键词： IL-6   WDFY1   luciferase   miRNA   transcription factor  

摘要： WD40 repeat and FYVE containing protein 1 (WDFY1) functions in membrane trafficking and protein complex scaffolding. WDFY1 has been studied in the immune system and in different oncogenic conditions. Therefore, comprehensive understanding of WDFY1 regulation mechanisms is much desired. In this study, we analyzed the promoter and 5'- and 3'-untranslated regions (UTRs) of and identified critical sequence elements, transcription factors (TFs), and miRNAs that collaboratively regulate gene expression. A 3.5 kb segment of the mouse promoter and 5'-UTR was cloned into a luciferase expression vector and transfected into HeLa cells. Luciferase assays of promoter deletion mutants revealed approximately four-fold increased activity attributed by a 500 bp distal fragment upstream of the 5'-UTR. Four TFs (Sp1, Ap-1, Hes1, and TCF7) were found to be critical for expression with binding sites spread throughout the promoter and 5'-UTR regions. Cloning of a 3.2 kb fragment of 3'-UTR into the luciferase expression vector led to an ~3.5-fold decrease in luciferase activity. Complementary siRNA and luciferase assays mutually confirmed our findings. Most importantly, IL-6, a critical cytokine in organ inflammation, was found to promote WDFY1 expression through the upregulation of Sp1 in primary renal mesangial cells. We, therefore, identified a potential inflammation-driven WDFY1 upregulation in mice.

关键词： Fourier transform infrared spectroscopy  

摘要： The current study uses the ionic liquid (IL) 1-butyl-3-methylimidazolium hexafluorophosphate, also known as BMIM-PF6, as a stabilizing agent to synthesize a bismuth oxide/ionic liquid/reduced graphene oxide (Bi2O3/IL/rGO) hybrid nanomaterial. A range of techniques, including scanning electron microscopy (SEM), X-ray diffraction (XRD), energy dispersive X-ray spectroscopy (EDX), Fourier transform infrared spectroscopy (FTIR), and UV-visible spectroscopy, were used to characterize the synthesized Bi2O3/IL/rGO nanocomposite. XRD confirmed that the composite was crystalline, and FTIR analysis suggested the presence of certain functional groups. The main elements oxygen, carbon, and bismuth were confirmed by EDX analysis, and SEM imaging showed an exfoliated and detached morphology. The synthesized nanocomposite was used to modify a glassy carbon electrode to develop a sensor for the detection of lead (Pb2+). The fabricated sensor was characterized using electrochemical impendence spectroscopy (EIS) and cyclic voltammetry (CV). The results indicated that the Bi2O3/IL/rGO/GCE was more conductive, having a charge transfer resistance (Rct) of 428.5 Omega compared to 1870 Omega for the bare electrode. The sensor has a low detection limit of 0.001 mu M and a quantification limit of 0.003 mu M. When tested on water and soil samples, the sensor was confirmed to have acceptable recovery rates, ranging from 95% to 102%. In conclusion, our fabricated sensor has excellent performance in terms of ease, affordability, energy usage, and quick efficiency.