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关键词： 外泌体   心肌细胞   miR-1275   IL-38   脓毒症  

摘要： 目的 探究心肌细胞来源的外泌体对脂多糖（LPS）诱导的脓毒症心肌细胞损伤的影响及其作用机制。方法 采用LPS构建脓毒症心肌细胞损伤模型，分离心肌细胞来源外泌体，与大鼠淋巴细胞共培养。实验分为以下2组：对照组（心肌细胞置于含10%胎牛血清的DMEM培养基中）和LPS组（心肌细胞置于含500 ng/mL LPS的培养基诱导3 h）。利用CCK-8检测淋巴细胞增殖情况；qRT-PCR和Western blotting法检测淋巴细胞中白细胞介素-38(IL-38)的mRNA和蛋白表达水平；通过生物信息学挖掘LPS诱导脓毒症心肌细胞损伤的关键外泌体，双荧光素酶报告验证其与IL-38是否有靶向关系。将LPS诱导的心肌细胞来源外泌体处理后的淋巴细胞与心肌细胞损伤模型共培养，实验分为以下2组：对照组（上层培养淋巴细胞，下层培养经LPS诱导的心肌细胞，共培养48 h）和LPS-exosome组（上层将淋巴细胞和经LPS诱导的心肌细胞来源外泌体混合培养，下层培养经LPS诱导的心肌细胞）。通过流式细胞术检测心肌细胞凋亡。并用人重组IL-38(Re-IL-38)蛋白处理LPS诱导的脓毒症心肌细胞损伤模型，实验分为以下2组：对照组（心肌细胞+LPS）和重组IL-38组（心肌细胞+IL-38预处理+LPS）。通过流式细胞术检测心肌细胞凋亡，Western blotting法检测心肌细胞中凋亡，PI3K/AKT通路相关蛋白表达水平。结果 与正常心肌细胞来源外泌体相比，LPS诱导的心肌细胞来源外泌体能够提高淋巴细胞增殖活力（P<0.05），且淋巴细胞中IL-38 mRNA和蛋白表达水平显著升高（P<0.05）。GEO数据库搜索及差异分析发现miR-1275为LPS诱导脓毒症心肌细胞损伤的关键外泌体；双荧光素酶报告基因结果显示，miR-1275 mimics增加WT-IL-38的荧光素酶活性（P<0.05）。LPS诱导的心肌细胞来源外泌体处理的淋巴细胞与LPS诱导的心肌细胞共培养，能抑制心肌细胞凋亡（P<0.05）。Re-IL-38处理后，心肌细胞凋亡率降低（P<0.05）,Bax蛋白表达量降低（P<0.05）,Bcl-2、p-PI3K和p-AKT蛋白表达量升高（P<0.05）。结论 LPS诱导的心肌细胞外泌体中的miR-1275介导上调淋巴细胞中IL-38的表达，激活PI3K/AKT通路从而抑制LPS诱导的心肌细胞凋亡。

关键词： 脑梗死   预后   生物标志物   细胞因子   微小核糖核酸   外泌体   综述  

摘要： 缺血性卒中（缺血性脑卒中）作为导致全球残疾和死亡的主要原因之一，近年来，其预后生物标志物的研究取得了显著进展。本研究综述了包括血细胞分析指标（如红细胞体积分布宽度）、ω-3多不饱和脂肪酸、细胞因子（如白介素6、肿瘤坏死因子α）、微小核糖核酸（miRNA，如miR-21、miR-155、miR-126）和外泌体在内的多种生物标志物在缺血性脑卒中中的作用机制及其作为预后评估的研究进展。研究结果显示，血细胞分析相关指标、ω-3多不饱和脂肪酸、白介素6、肿瘤坏死因子α的浓度与缺血性脑卒中的严重程度和预后密切相关。其中，较高基线红细胞体积分布宽度与复发性缺血性脑卒中的风险增加相关，并且与缺血性脑卒中的复发时间呈负相关。特定miRNA如miR-21通过抑制细胞凋亡和促进神经元存活，改善神经功能恢复；而miR-155通过调控炎症反应加剧脑损伤，其表达水平可预测缺血性脑卒中的复发；此外，miR-126在血管生成和神经保护中发挥关键作用。外泌体则通过携带抗炎因子、神经营养因子、抗氧化酶和热休克蛋白等，显著减轻缺血再灌注引起的炎症和损伤，从而影响缺血性脑卒中的预后。尽管这些生物标志物在缺血性脑卒中预后评估中显示出巨大潜力，但其临床应用仍面临诸多挑战，包括个体间差异、长期效应和安全性研究不足以及技术标准化问题。未来研究应进一步探讨这些标志物的作用机制，开发标准化的检测方法，并进行大规模临床验证，以期将其应用于临床，改善缺血性脑卒中患者的预后和生活质量。

关键词： 慢性阻塞性肺疾病   C反应蛋白   白细胞介素-6   血常规   诊断准确率  

摘要： 探索C反应蛋白(CRP)、白细胞介素-6(IL-6)联合血常规检测在慢性阻塞性肺疾病(COPD)诊断中的实际应用价值,旨在为临床提供更高效的诊断路径,提升早期识别的准确性与时效性。方法 研究对象为56例明确诊断为COPD的患者。依据检测方案差异,随机分为两组。对照组采用传统血常规检测方式进行评估,样本量为28例。实验组在常规血常规基础上,增加CRP与IL-6指标检测,同样纳入28例患者。两组分别接受标准临床评估流程,并记录诊断正确率、相关炎症指标水平,以及与疾病严重程度之间的相关性。结果 数据显示,实验组的整体诊断准确率达到92.9%。相比之下,对照组的准确率为75.0%。两者之间存在统计学显著差异(P<0.05)。进一步分析发现,CRP与IL-6水平在COPD不同分期中表现出逐步升高趋势,与病情程度密切相关。特别是在中重度及急性加重期患者中,这两个炎症因子呈明显高表达状态。此外,实验组在判断疾病阶段及并发炎性反应方面更具参考价值,为病情评估提供了更直观的量化指标。结论 单一血常规检测在临床早期识别COPD方面存在局限。将CRP、IL-6纳入检测体系后,不仅提高了诊断准确性,也强化了对炎症反应强度的判断能力。该联合方案有助于在症状不典型或初期表现不明显的患者中进行更明确的识别,避免漏诊与误判。同时,炎症因子水平与疾病严重程度之间的关联性也提示其在病程管理与预后评估中具备一定价值。未来应进一步拓展样本量,并结合肺功能指标,方法 在分级诊疗体系中的实用性及推广可行性。

关键词： 外泌体   心肌细胞   miR-1275   IL-38   脓毒症  

摘要： 目的 探究心肌细胞来源的外泌体对脂多糖（LPS）诱导的脓毒症心肌细胞损伤的影响及其作用机制。方法 采用LPS构建脓毒症心肌细胞损伤模型，分离心肌细胞来源外泌体，与大鼠淋巴细胞共培养。将实验分为以下2组：对照组（心肌细胞置于含10%胎牛血清的DMEM培养基中）和LPS组（心肌细胞置于含500 ng/mL LPS的培养基诱导3 h）。利用CCK8检测淋巴细胞增殖情况；qRT-PCR和Western blotting法检测淋巴细胞中白细胞介素-38(IL-38)的mRNA和蛋白表达水平；通过生物信息学挖掘LPS诱导脓毒症心肌细胞损伤的关键外泌体，双荧光素酶报告验证其与IL-38是否有靶向关系。将LPS诱导的心肌细胞来源外泌体处理后的淋巴细胞与心肌细胞损伤模型共培养，将实验分为以下两组：对照组（上层培养淋巴细胞，下层培养经LPS诱导的心肌细胞，共培养48 h）和LPS-exosome组（上层将淋巴细胞和经LPS诱导的心肌细胞来源外泌体混合培养，下层培养经LPS诱导的心肌细胞）。通过流式细胞术检测心肌细胞凋亡。并用人重组IL-38(Re-IL-38)蛋白处理LPS诱导的脓毒症心肌细胞损伤模型，将实验分为以下两组：对照组（心肌细胞+LPS）和重组IL-38组（心肌细胞+IL-38预处理+LPS）。通过流式细胞术检测心肌细胞凋亡，Western blotting法检测心肌细胞中凋亡，PI3K/AKT通路相关蛋白表达水平。结果 与正常心肌细胞来源外泌体相比，LPS诱导的心肌细胞来源外泌体能够显著提高淋巴细胞增殖活力（P<0.05），且淋巴细胞中IL-38 mRNA和蛋白表达水平显著升高（P<0.05）。GEO数据库搜索及差异分析发现miR-1275为LPS诱导脓毒症心肌细胞损伤的关键外泌体；双荧光素酶报告基因结果显示，miR-1275 mimics显著增加WT-IL-38的荧光素酶活性（P<0.05）。LPS诱导的心肌细胞来源外泌体处理的淋巴细胞与LPS诱导的心肌细胞共培养，能抑制心肌细胞凋亡（P<0.05）。Re-IL-38处理后，心肌细胞凋亡率降低（P<0.05）,Bax蛋白表达量降低（P<0.05）,Bcl-2、p-PI3K和p-AKT蛋白表达量升高（P<0.05）。结论 LPS诱导的心肌细胞外泌体中的miR-1275介导上调淋巴细胞中IL-38的表达，激活PI3K/AKT通路从而抑制LPS诱导的心肌细胞凋亡。

关键词： canine parvovirus   VP2   IL-12   baculovirus expression system  

摘要： Background: Canine parvovirus (CPV) is a highly pathogenic virus that predominantly affects puppies, with mortality rates exceeding 70%. Although commercial multivalent live attenuated vaccines (MLV) are widely employed, their efficacy is often compromised by maternal antibody interference. Consequently, the development of novel vaccines remains imperative for effective CPV control. Methods: Recombinant CPV VP2 protein (rVP2) and canine interlukine 12 protein (rcIL-12) were expressed using the Bac-to-Bac baculovirus expression system and the biological activity of these proteins was assessed through hemagglutination, Cell Counting Kit-8 (CCK8) and IFN-gamma induction assays. The combined immunoenhancement effect of rVP2 and rcIL-12 protein was evaluated in puppies. Results: Both rVP2 and rcIL-12 were successfully expressed and purified, exhibiting confirmed antigenicity, immunogenicity, and bioactivity. Co-administration of rVP2 with rcIL-12 elicited higher neutralizing antibody titer (6-7 times higher), complete challenge protection efficiency (no clinical symptoms and tissue and organ lesions), fewer viral shedding (decreasing significantly 8-day post challenge) and superior viral blockade (lower viral load in the organism) compared to rVP2 alone. Conclusions: Our findings demonstrate that rVP2 co-administered with rcIL-12 induces robust protective immunity in puppies and significantly mitigated the inhibitory effects of maternal antibodies. This represents a promising strategy for enabling earlier vaccination in puppies and rational design of CPV subunit vaccines.

摘要： Dermal resident memory CD4+ T cells (dTrm) provide protection against vector-borne infections. However, the factors that promote their development remain unclear. We tested if an mRNA vaccine, encoding a protective leishmanial antigen, induced dTrm cells. The mRNA vaccine induced robust systemic T-cell responses, but few Trm cells were found in the skin. Since IL-12 promotes Th1 responses, we tested whether IL-12 mRNA combined with the mRNA vaccine could enhance dTrm cell development. This combination significantly expanded Leishmania-specific Th1 cells expressing skin-homing molecules and memory T cell markers in the draining lymph node. Additionally, higher numbers of dTrm cells were maintained in the skin, and mice exhibited functional immunity indicated by a delayed hypersensitivity response and protection upon challenge with Leishmania. These findings highlight IL-12 as a key driver of CD4+ dTrm development, enabling their global seeding across the skin, and underscore the potential of IL-12-enhanced mRNA vaccines to generate durable immunity against cutaneous leishmaniasis and other skin-targeted infections.

关键词： IL-2 epitopes   Cancer immunotherapy   Large Language models   MERCI, machine learning techniques  

摘要： Interleukin-2 (IL-2) based immunotherapy has been approved for treating certain types of cancer, as IL-2 plays a crucial role in regulating the immune system. In this study, we developed a method for predicting IL-2-inducing peptides. Our method was trained, tested, and validated on a main dataset containing 6,574 experimentally validated Major histocompatibility complex (MHC) binders, including 3,429 IL-2-inducing and 3,145 non-inducing peptides. A primary analysis of IL-2 inducing and non-inducing peptides revealed that certain residues, such as alanine and leucine, are more abundant in IL-2-inducing peptides. Initially, we developed alignment-based methods, which demonstrated high precision but limited coverage. Subsequently, we developed artificial intelligence-based models, including machine learning (ML), deep learning (DL), and large language models (LLM), to predict IL-2-inducing peptides. Our Extra Tree-based model, developed using dipeptide composition and peptide length, achieved a maximum AUC of 0.82. Finally, we constructed ensemble models that combined artificial intelligence and alignment-based methods. Our best ensemble model, which integrates the Extra Tree-based model with MERCI, achieved the highest AUC of 0.84 and an MCC of 0.51 on the main dataset. One limitation of the main dataset is that both IL-2-inducing and non-inducing peptides are MHC binders. To address this limitation, we created two additional datasets: Alternate Dataset 1, consisting of 3,429 IL-2-inducing peptides and 3,429 non-inducing peptides (MHC non-binders), and Alternate Dataset 2, consisting of 3,429 IL-2-inducing peptides and 3,439 non-inducing peptides (MHC binders + MHC non-binders). Our best ensemble model achieved AUCs of 0.9 and 0.8 with MCCs of 0.61 and 0.44 on Alternate Datasets 1 and 2, respectively. To assist the scientific community, we have integrated the best models from this study into a standalone software and web server, IL2pred, which enables users to predic

关键词： transplantation   regulatory T cells   germinal center   donor-specific antibodies   humoral tolerance  

摘要： Antibody-mediated rejection (ABMR) caused by donor-specific Abs (DSAs) is still the leading cause of late graft loss following clinical organ transplantation, and effective strategies to combat ABMR are still elusive. We previously showed that rIL-2 complexed with anti-IL-2 mAb clone JES6-1A12 (IL-2 cplx) leads to the selective expansion of regulatory T cells (Tregs) and the prolonged survival of MHC-mismatched skin allografts. Although the grafts were eventually rejected, mice failed to develop DSAs. Here, we investigated the impact of IL-2 cplx on the humoral response and germinal center (GC) reaction during allograft rejection. IL-2 cplx treatment prevents Bcl-6 upregulation, leading to suppressed development of GC T and B cells. The IL-2 cplx-induced impairment of GC development limits IgG allo-Ab production but allows for IgM synthesis. By employing a hapten-carrier system to investigate affinity maturation, we found that IL-2 cplx induces a distinct shift in specific Ab production favoring low-affinity IgM while simultaneously decreasing IgG responses. These findings illuminate the potential of IL-2 cplx therapy for inducing humoral tolerance, potentially paving the way for refining strategies aimed at preventing and treating ABMR.

关键词： IL-27   HBV persistence   CD8 T cell dysfunction   Liver microenvironment  

摘要： Background Persistent chronic hepatitis B virus (HBV) infection leads to chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. The immunosuppressive tissue microenvironment in the liver restricts the immune response, potentially facilitating persistent HBV infection. This study examined the expression of immunity-related factors in the liver in response to HBV. Methods We performed gene expression profiling on mice subjected to HBV DNA hydrodynamic transfection to identify transcriptomic changes. The expression of IL-27 was validated through Western blotting, ELISA, and immunohistochemical staining. Liver macrophages in mice were depleted using clodronate-liposomes to evaluate their role in IL-27 production. IL-27 knockout mice were generated to examine the effects of IL-27 deficiency on CD8 T cell dysfunction and HBV persistence. Results Transcriptomic analysis demonstrated that IL-27 is significantly induced in the liver in response to HBV DNA. The elevated levels of IL-27 are strongly correlated with HBV persistence and are linked to CD8 T cell dysfunction, characterized by increased expression of PD-1 and Tim-3, along with reduced IFN-γ production in liver-infiltrating T cells. Furthermore, depleting macrophage-lineage cells using clodronate-liposomes significantly reduces IL-27 production in the liver and promotes viral clearance. Additionally, mice with IL-27 deficiency exhibit enhanced HBV clearance and restored CD8 T cell function. Conclusions Collectively, IL-27 is significantly induced by HBV in the liver, and its production is strongly associated with HBV persistence and CD8 T cell dysfunction. This highlights the immunosuppressive role of IL-27 in the liver microenvironment and suggests that IL-27 could serve as a potential therapeutic target for HBV infection.

关键词： NSCLCs   tumor-derived small extracellular vesicles (TEVs)   IL-1 beta   cancer progression   EMT   immune suppression  

摘要： Background: Increased IL-1 beta levels may promote carcinogenesis and metastasis by affecting tumor biology and the tumor microenvironment (TME). In this context, extracellular vesicles (EVs) play a key role in cell-to-cell communication, thus modulating the TME and immune response. Here, we aimed to test whether tumor-derived small EVs (TEVs) isolated from sensitive and osimertinib-resistant (OR) non-small-cell lung cancer (NSCLC) cells may promote EMT via fibronectin binding to alpha 5 beta 1 integrin as well as suppress the immune system and if these effects may be favored by IL-1 beta. Methods: TEVs were isolated from control, OR, and IL-1 beta-stimulated NSCLC cells. Expressions of fibronectin and PD-L1 were screened in TEVs and the mRNA levels of vimentin and SMAD3 were also assessed in cancer cells after TEV co-culturing. Furthermore, to detect the effect on immune cells, we co-cultured TEVs with lung cancer patients' peripheral blood mononuclear cells (PBMCs). Results: TEVs were positive for fibronectin and the highest protein levels were found in TEVs obtained from the OR and IL-1 beta-stimulated cells. TEV-mediated activation of alpha 5 beta 1 signaling led to the upregulation of vimentin and SMAD3 mRNA in NSCLC cells and stimulated cell migration. EVs also increased PD-1, CTLA-4, FOXP3, TNF-alpha, IL-12, and INF-gamma mRNA in lung cancer patients' immune cells. Conclusions: Our findings indicate that TEVs promote EMT in NSCLC cells by the activation of the fibronectin-alpha 5 beta 1 axis. Finally, IL-1 beta stimulation induces TEV release with biological properties similar to OR TEVs, thus leading to cancer invasion and immune suppression and suggesting that inflammation can promote tumor spreading.