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摘要： Bullous pemphigoid (BP) is a subepidermal autoimmune blistering disease characterized by erythema, tense blisters, pruritus, and pain. The pathogenesis of BP involves antibodies targeting structural proteins at the dermal-epidermal junction. Prior studies have identified IL-17 as a potential therapeutic target in BP. We hypothesize IL-17 blockade with ixekizumab, an IL-17 A inhibitor, may have a targeted, disease-modifying effect on BP. We conducted a single-center, exploratory, open-label, single-arm pilot study using ixekizumab treatment for 12 weeks. Treatment response was assessed by new blister formation, Bullous Pemphigoid Disease Area Index (BPDAI), and use of prednisone rescue. A total of 4 patients were recruited, with 3 withdrawing during the treatment phase with worsening disease. All 3 withdrawn patients required oral prednisone rescue therapy. One patient completed the study with a 60% improvement in blister/erosion BPDAI on ixekizumab and rescue oral prednisone. RNA levels in baseline disease tissue and control samples were analyzed using a NanoString® panel of 249 inflammatory genes. Cytokine analysis was performed on serum and blister fluid for interleukin (IL)-1b, -2, -4, -5, -6, -12p70, -17a, -18, -21, -22. Array-based ELISAs were used for transforming growth factor beta (TGF-β) and matrix-metalloprotease-2 (MMP-2), MMP-9, and MMP-13 expression. There were 24 statistically significant differentially expressed genes (DEGs) identified. Type I and II IFNs as well as the Th2 chemokine CCL13 were upregulated in BP. Serum pre-treatment IL-17 A was elevated in 1 patient and was no longer detectable after week 12. Cytokine analysis demonstrated elevated serum IFN-γ, MMP-13, and Th2 cytokines in BP patients after receiving ixekizumab relative to pre-treatment levels, with a decrease in MMP-2. Our clinical data, RNA, and cytokine analysis suggest that IL-17 A inhibition with ixekizumab may not target the critical pathways in BP and support Typ

关键词： 小鼠细小病毒   单克隆抗体   双抗体夹心ELISA   抗原检测  

摘要： 制备小鼠细小病毒（MVM）VP2蛋白单克隆抗体，建立一种基于双抗体夹心ELISA的实验动物小鼠MVM抗原检测方法。应用原核表达质粒表达VP2蛋白，纯化蛋白免疫雌性BALB/c小鼠制备单克隆抗体，以获得的单抗1C5作为捕获抗体，HRP标记的单抗8B9作为检测抗体，优化条件建立双抗体夹心ELISA检测方法，并对其灵敏性、特异性和重复性进行系统评价并与qPCR检测进行比对分析。结果显示，表达纯化了MVM VP2 蛋白，免疫雌性BALB/c小鼠，免疫程序结束后筛选获得2株单克隆抗体，命名为1C5和8B9，Western-blot和间接免疫荧光鉴定发现这2株抗体可与MVM病毒和重组蛋白特异性结合。以1C5作为捕获抗体，HRP标记的8B9作为检测抗体建立了双抗体夹心ELISA方法，其对MVM病毒和重组蛋白最低检测限分别为103.4TCID50/mL和50 ng/mL；与4种常见病毒感染的小鼠血清均无交叉反应，包被ELISA板条的批间变异系数均小于10%，与qPCR检测结果相比总符合率为100%。结果表明，成功制备了2株MVM单克隆抗体，应用这2株抗体建立的MVM双抗体夹心ELISA方法具有较好的灵敏性和特异性，与qPCR检测法相比一致性较好，为实验动物小鼠的临床诊断、高通量筛查监测提供了有效的检测方法，从而保障实验动物的SPF状态，为生命科学研究提供健康可靠的实验动物。

关键词： Acute lymphoblastic leukemia   Cancer therapy   Computer modeling   Enzyme   Immunogenicity   Protein cross-linking   Protein structure   Protein-protein interaction  

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关键词： 3-Indolepropionic acid   Dose-weighted network pharmacology   Gastric cancer   Ginsenoside Rh2   Gut microbiota   Systems biology   TNF  

摘要： INTRODUCTION:Gastric cancer (GC) remains a formidable global health issue with limited therapeutic options. ShenXia KuanZhong Decoction (SXKZD), a classical traditional Chinese medicine (TCM) formula, is used to manage GC; however, its anti-tumor mechanisms remain poorly understood. MATERIAL AND METHODS:The anti-GC effects of SXKZD were investigated in a GC model using dose-weighted network pharmacology, molecular docking, molecular dynamics (MD) simulations, and pharmacokinetic profiling. Its impacts on tumor metabolism, immunity, and gut microbiota were assessed. A gut microbiota-substrate-metabolite (GM-S-M) network was constructed, and key targets and pathways were analyzed using computational and experimental methods. RESULTS:SXKZD treatment significantly alleviated tumor progression in GC models. Network analysis revealed upregulated TNF and IL6 expression in GC, which SXKZD reduced, alongside enrichment in IL-17 and TNF signaling pathways. Molecular docking and MD simulations confirmed stable binding of Ginsenoside Rh2 and 3-Indolepropionic acid to TNF, with binding energies of -147.63 kJ/mol and - 98.63 kJ/mol, respectively. Pharmacokinetic profiling showed 3-Indolepropionic acid's high bioavailability, while GM-S-M analysis identified key microbial taxa (e.g., Lactobacillus plantarum, Akkermansia muciniphila) modulated by SXKZD, enhancing anti-tumor immunity and *** further confirm these computational predictions, in vitro CCK-8 assays revealed that GRh2 and IPA inhibited AGS cell growth in a concentration-dependent manner, with IC values of 68.74 ± 1.27 µg/mL and 780.60 ± 24.40 µg/mL at 24 hours, respectively. Western blot analysis demonstrated that GRh2 more effectively suppressed TNFα expression, whereas CETSA showed that IPA provided superior thermal stabilization of TNFα. CONCLUSIONS:SXKZD mitigates GC by modulating gut microbiota and inhibiting TNF signaling, offering a mechanistic basis for its therapeutic potential in GC management.

关键词： Hyperbaric oxygen   Inflammatory cytokines   Spinal cord injury   γδT cell  

摘要： BACKGROUND:Hyperbaric oxygen (HBO) treatment can attenuate the inflammatory response after spinal cord injury (SCI), but the specific molecular mechanisms are unclear. γδT cells have emerged as key immune regulatory cells. This study aims to analyze the mechanism of HBO treatment reducing SCI mainly from the perspective of γδT cells. METHODS:Contusion SCI models were established in WT, TCRδ, and IL-17 mice, and HBO treatment was performed. Hindlimb locomotor function, ChAT-positive motor neuron numbers, γδT cell proportion, and inflammatory cytokines level of different experimental groups were compared after SCI. Furthermore, we measured the protein and gene expression of STAT3 and RORγt in both WT mice and STAT3 overexpression mice following HBO treatment. RESULTS:Our results showed γδT cells, especially IL-17 + γδT cells, were involved in SCI. HBO treatment improved locomotor recovery, reduced the proportion of γδT and IL-17 + γδT cells, and decreased inflammatory cytokines in WT mice after SCI. However, HBO treatment did not affect locomotor recovery or inflammatory cytokines level in TCRδ or IL-17 mice after SCI. In addition, HBO treatment significantly inhibited the expression of STAT3 and RORγt in WT mice after SCI. Over-expression of STAT3 weakened the inhibitory effect of HBO on IL-17 + γδT cells and inflammatory cytokines after SCI. CONCLUSIONS:These findings indicated that IL-17 + γδT cells may play a key role in HBO treatment alleviating the inflammatory response after SCI. HBO treatment may regulate IL-17 + γδT cells by modulating the pathway of STAT3/RORγt. This study provides the theoretical basis and potential therapeutic target for HBO treatment in SCI.

关键词： Disease management   IL-6 inhibition   juvenile idiopathic arthritis   pediatric rheumatology   treatment   unmet needs  

摘要： INTRODUCTION:Medical management of juvenile idiopathic arthritis (JIA) presents a significant challenge in pediatric rheumatology. Ideally, the treatment target is remission, though achieving this remains complex. Interleukin-6 (IL-6) inhibitors (IL-6is) play an important role, targeting the inflammatory pathways central to JIA pathogenesis. However, their optimal use is debated. AREAS COVERED:This narrative review examined JIA care needs and IL-6 inhibition. A SPIDER-based literature search of PubMed/MEDLINE, Semantic Scholar, WorldCat, Cochrane Library, Embase, CINAHL, ICTRP, and *** (to May 2025), identifying 56 studies from 246 records published between 2018-2025. Key unmet needs include difficulty controlling the disease, diagnostic delay, shortcomings in biomarker research, and multidisciplinary support. Tocilizumab, a well-studied IL6i, showed efficacy in symptom reduction, disease control, and reduced glucocorticoid use. EXPERT OPINION:Addressing gaps in JIA management, such as delayed diagnosis and inadequate disease control, is essential. Experts advocate for early IL6i use within a treattotarget framework, optimizing outcomes and minimizing glucocorticoid use. Recognizing benefits for highrisk JIA subtypes, experts support earlier tocilizumab integration into treatment algorithms, offering valuable options for refractory oligoarthritis and uveitis. Ultimately, bridging gaps in JIA management and reshaping real-world outcomes hinges on integrating clinical insight and research outcomes - a process driven by precision medicine.

关键词： MDA5   interferon   juvenile dermatomyositis   muscle   myositis-specific auto-antibodies  

摘要： OBJECTIVES:This study aimed to establish the role of myositis-specific antibodies (MSAs) in the association between type I interferon (IFN-I) plasma levels and disease activity in juvenile dermatomyositis (JDM). METHODS:We prospectively obtained 400 samples from 101 JDM patients from 2 independent cohorts. Autoantibody levels were determined for all patients. Muscle activity was assessed using the Childhood-Myositis Assessment Scale (CMAS). Two characterized homebrew digital ELISAs measuring respectively, all 12 IFN-alpha subtype proteins (IFN-α), and IFN-beta (IFN-β) were used to quantify IFN-I in patient plasma. Receiver operating characteristic (ROC) curve analysis was used to identify IFN-I thresholds associated with CMAS changes. Correlations between IFN-I levels and CMAS were assessed at recruitment using Spearman's test, and longitudinally using mixed-effects models to account for repeated measures. RESULTS:IFN-α levels were higher in melanoma differentiation-associated gene 5 (MDA5)-positive patients while IFN-β levels were comparable across MSA subgroups. IFN-β was found to be more effective than IFN-α in distinguishing between active and inactive muscle disease, and between severe and non-severe disease status. Over the disease course, we identified IFN-β as a reliable biomarker of muscle disease activity regardless of MSA expression. In contrast, IFN-α levels showed a specific association with CMAS only MDA5-positive patients. CONCLUSIONS:This exclusive association of IFN-α levels with muscle clinical score in anti-MDA5-positive patients suggests a subgroup-specific pathophysiological mechanism. These findings underscore potentially distinct roles for IFN-I subtypes (IFN-α and IFN-β) as circulating biomarkers of muscle disease activity in JDM according to the MSA expression.

摘要： NK cell-based immunotherapy of solid tumors has been shown to be increasingly successful, but much effort is still needed to optimize its efficacy. This study explores the effects of treatment with low, non-toxic doses of IFN-γ and TNF-α on the susceptibility of breast cancer (BC) cell lines (MCF-7, MDA-MB-231, and MDA-MB-468) cultured in 2D and 3D as spheroids, to NK cell-mediated antitumor function. We evaluated the expression of (i) ligands for NK cell-activating receptors on BC cells, (ii) death and adhesion molecules on BC cells, and (iii) the expression of NK cell-receptors on NK cells infiltrating BC spheroids. Cytokine treatment significantly increased the expression of FAS, TRAIL-R2, and ICAM-1 in all BC cell lines, enhancing NK cell-mediated apoptosis and promoting NK cell-tumor cell conjugate formation. Differently, the expression of ligands for activating receptors remained essentially unchanged. In BC spheroids, the treatment with IFN-γ and TNF-α enhanced NK-cell infiltration, with increased expression of activating receptors (NKG2D, DNAM-1, NKp30, and NKp46) on infiltrating NK cells. However, regardless of treatment, markers of NK cell exhaustion, such as PD-1 and CTLA-4, were also upregulated, the latter especially in triple-negative BC (TNBC) MDA-MB-231 spheroids, thus suggesting an exhausted phenotype of NK cells infiltrating spheroids despite activation. Cytokine treatment resulted in a significant NK cell-mediated reduction in spheroid size, accompanied by an increased apoptotic state, with effects more pronounced in MCF-7 than in MDA-MB-231 spheroids. These results indicate that, although the treatment with IFN-γ and TNF-α improves NK cell-mediated tumor interaction and apoptosis, its therapeutic efficacy may be dependent on the BC subtype, with TNBC spheroids showing greater resistance. These findings highlight the importance of the tumor microenvironment (TME) in shaping NK cell responses and suggest that combining IFN-γ and TNF-α treatments wi

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