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关键词： 鸡   早幼粒细胞白血病蛋白   单克隆抗体   定位   表达分布  

摘要： 研究旨在研制抗鸡源早幼粒细胞白血病蛋白（PML）的单克隆抗体。试验采用PCR和同源重组技术构建鸡PML基因的1 228～1 566 nt原核表达质粒，命名为pGEX-6p-1-PML-P2。使用IPTG诱导表达后，以纯化成功的GST-PML-P2蛋白免疫小鼠，将小鼠脾细胞与骨髓瘤细胞SP2/0融合，通过间接免疫荧光（IFA）方法筛选出阳性细胞克隆；用激光共聚焦、Western blot及免疫沉淀（IP）验证单克隆抗体的特性。结果显示：成功获得3株能够分泌抗鸡PML单克隆抗体的杂交瘤细胞株，命名为4C2、3F4和4G3;3株单克隆抗体的亚类均为IgG1，其腹水的IFA效价分别为1∶6 400(4C2)、1∶12 800(3F4)和1∶25 600(4G3)；单克隆抗体4C2可特异性检测禽源细胞（HD11、MSB-1、LMH和DF-1）及非禽源细胞（MDCK、MA104和293T）中的内源性PML蛋白，激光共聚焦可观察到DF-1细胞内源性PML蛋白在细胞核内呈点状分布；3株单克隆抗体均能有效免疫沉淀外源高表达的鸡PML蛋白且单克隆抗体4C2能有效免疫沉淀DF-1细胞内源性的PML蛋白，确定PML在鸡的心脏、肝脏、脾脏、肺脏、肾脏、胸腺及法氏囊组织中广泛表达。研究表明，成功制备的3株抗鸡PML单克隆抗体可以有效检测不同细胞系及鸡组织中的PML蛋白并免疫沉淀细胞中内源及外源表达的PML蛋白，为深入探究鸡PML蛋白的功能及其在禽类疾病防控中的应用提供了不可或缺的工具。

关键词： Cancer immunotherapy   Coagulation   Immunology   Oncology   Platelets   Vascular biology  

摘要： Malignancies increase the risk for thrombosis and metastasis dependent on complex interactions of innate immune cells, platelets, and the coagulation system. Immunosuppressive functions of platelets and macrophage-derived coagulation factors in the tumor microenvironment (TME) drive tumor growth. Here, we show that patients with malignancies and tumor-bearing mice have increased levels of coagulation factor (F) X-expressing circulating monocytes engaged in platelet aggregate formation. This interaction and resulting thrombin generation on platelets interferes with monocyte differentiation and antigen uptake of antigen-presenting cells (APCs). Myeloid cell-specific deletion of FX or abrogated FXa signaling via protease activated receptor 2 (PAR2) averts the suppressive activity of platelets on tumor cell debris uptake and promotes the immune stimulatory activity of APCs in the TME. Myeloid cell FXa-PAR2 signaling deficiency specifically enhances activation of the cGAS-STING-IFN-I pathway with a resulting expansion of antigen oral anticoagulants expands T cell priming-competent immune cells in the TME and synergizes with the reactivation of exhausted CD8+ T cells by immune checkpoint inhibitors for improved antitumor responses. These data provide mechanistic insights into the emerging clinical evidence demonstrating the translational potential of FXa inhibition to synergize with immunotherapy.

关键词： inflammation   cell-junction dynamics   actin dynamics   JAIL   stress fibers   contractility   VE-cadherin dynamics   shear stress  

摘要： Inflammatory endothelial phenotypes describe distinct cellular patterns essential for controlling transendothelial migration of leukocytes (TEM). While TNF-alpha-induced CAM expression mediates leukocyte interaction, the role of a potential inflammatory morphological phenotype (IMP) - characterized by barrier-function decrease and shape-change in TEM - remains unclear. This study identifies the TNF-alpha-induced IMP as indispensable for neutrophil TEM, while regulating barrier-function. The TNF-alpha-induced IMP progresses through two states: an intermediate state that transiently enhances barrier function via MLC-dephosphorylation, junctional actin recruitment and VE-cadherin linearization, protecting the monolayer from collapse;while the subsequent development of the IMP requires MLC rephosphorylation, junctional actin disassembly, stress fiber formation and Arp2/3-mediated membrane protrusions causing shape-change. This in turn dilutes junctional VE-cadherin, forming intercellular gaps for neutrophil TEM, while inducing junction-associated intermittent lamellipodia (JAIL) to locally restore VE-cadherin adhesion, appearing as gap/JAIL cycles driving junctional dynamics. VE-cadherin overexpression blocks TNF-alpha-induced IMP and gap/JAIL cycling, reducing TEM by similar to 80% without altering CAM expression. These findings highlight gap/JAIL cycling and MLC phosphorylation as key IMP regulators and potential therapeutic targets for inflammatory diseases.

关键词： inflammasome   NLRP1   gestational malaria   Plasmodium falciparum   Plasmodium vivax   trophoblasts  

摘要： We hypothesized that variants in inflammasome-related genes could influence susceptibility to gestational malaria (GM). To test this, we conducted an association study in a cohort of pregnant women from a malaria-endemic region in northern Brazil, assessing whether specific functional single nucleotide variants (SNVs) in inflammasome genes affect (1) the response to Plasmodium infection and (2) the development of placental malaria. Our findings revealed that the NLRP1 ***1154Val variant was associated with a protective effect against Plasmodium infection. Moreover, IL1B SNVs appeared more prevalent in severe cases. Additionally, multivariate analyses incorporating placental blood cytokines, growth factors, and immunohistochemical features revealed that the NLRP1 ***1154Val variant correlated with a healthier placental state, highlighting a potential protective role of the NLRP1 inflammasome in GM. For the first time, we showed that infected red blood cells induce NLRP1- and caspase-1-dependent pyroptosis in BeWo trophoblast cells, identifying a novel inflammasome pathway involved in GM pathogenesis. Our study identifies a genetic variant underlying NLRP1 contribution to GM and suggests that NLRP1 may be an under-explored inflammasome receptor in malaria and infected erythrocytes' sensing. (c) 2025 The Author(s). The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

关键词： IL-22   claudins   inflammatory-triggered abortion   pregnancy loss   tight junctions  

摘要：

关键词： Small bowel Crohn's disease (CD   Terminal ileal (TI) stricture diagnosis   Magnetic resonance enterography (MRE)   Computed tomography enterography (CTE)  

摘要： PurposeCrohn's disease (CD) is characterized by enteric inflammation, often resulting in strictures and penetrating complications, which may alter patient management prior to the initiation of biologic therapy. Our aim is to assess the frequency of missed stricturing and internal penetrating complications in CD patients on computed tomography enterography (CTE) and magnetic resonance enterography (MRE) performed prior to anti-TNF *** retrospectively reviewed patients from two tertiary centers who underwent CTE\MRE within six months before starting anti-TNF therapy. Patients with prior intestinal surgery, anti-TNF therapy, or radiology reports indicating strictures or penetrating complications were excluded. Two GI radiologists re-evaluated CTE\MRE images for active inflammation, small bowel strictures (SAR/AGA/SPR criteria), and internal penetrating complications. Senior radiologists reviewed CTE/MRE exams with missed findings and identified potential causes of error. Descriptive statistics were used to summarize the prevalence of missed findings. Where applicable, comparisons were made using chi-square ***250 CD patients were included. 239 (96%) exhibited active terminal ileal (TI) inflammation by expert review of CTE\MRE images. Missed TI strictures were identified in 56 patients (22%). 16 patients (6.4%) had unreported penetrating complications, including ileoappendiceal fistulas (n = 6), sinus tracts (n = 6), complex fistulas (n = 4), inflammatory masses (n = 3), ileocecal fistula (n = 1), and abscess (n = 1). There were no significant differences between institutions in the rates of strictures (18/69 vs. 38/181, P = 0.4) or penetrating complications (5/69 vs. 11/181, P = 0.77). 46% of the strictures appeared obvious upon re-evaluation by senior radiologists, potentially implicating a failure to measure associated small bowel dilation. 38% of missed strictures had multifocal areas of luminal narrowing, and maximal proximal dilation was loc

关键词： multiple myeloma   multi-omics atlas   non-immune cells   signaling pathway   metabolic reprogramming-targeted  

摘要： Multiple myeloma (MM) is highly heterogeneous, with relapse occurring in the majority of cases, and recent advancements in single-cell RNA sequencing (scRNA-seq), sc-metabolism profiling, and bulk RNA-seq have facilitated the identification of cell subpopulations and metabolic reprogramming at the single-cell level, uncovering novel molecular mechanisms. This study aims to establish a multi-omics atlas of MM, characterizing the cell subpopulations and signaling pathways that drive immune evasion and disease progression. Additionally, sc-metabolic profiling identifies reprogramming patterns and informs therapeutic screening. We integrated scRNA-seq and bulk RNA-seq data using R to analyze immune and non-immune cell features and pathways in MM. Metabolic reprogramming was assessed via sc-metabolic profiling, and drug candidates were screened through multi-omics integration, with efficacy evaluated in vitro using CCK-8 assays, flow cytometry, Western blotting, and CalcuSyn software. Novel MM subpopulations were identified, including myeloma-activated hematopoietic stem cells and ISG15+ B cells, which correlated with survival and were validated by multiplex immunofluorescence. IFN-gamma is primarily secreted by effector memory CD8+T cells, and IFN-alpha is primarily secreted by non-classical monocytes, driving an IFN-gamma/alpha-B2M feedback loop. Multi-omics identified four drug candidates, each demonstrating anti-tumor effects against myeloma cell lines.

关键词： chronic hepatitis B   serum ferritin   HBsAg   PEG IFN-alpha 2b   ALT  

摘要： Background: Chronic hepatitis B (CHB) caused by HBV infection leads to persistent liver inflammation. PEG IFN alpha-2b shows higher functional cure rates than oral antivirals, but identifying optimal candidates remains challenging. This study evaluates serum ferritin as a predictive biomarker for PEG IFN alpha-2b efficacy. Methods: Serum ferritin, HBsAg and transaminase were monitored in CHB patients treated with PEG IFN alpha-2b at baseline (0w), 12w and 24w. Patients were divided into the effective group (HBsAg reduction >= 1 log10 IU/mL or clearance) and the ineffective group (HBsAg reduction <1 log10 IU/mL) at 24 weeks. Similarly, for the results at 48 weeks, the patients were divided into the clearance group (achieving HBsAg clearance) and the non-clearance group (not achieving HBsAg clearance). To analyze the correlation between serum ferritin and the decrease of HBsAg, a multivariate logistic regression model was constructed. Potential confounding factors such as age, gender, HBeAg and HBV DNA status, ALT, AST, and iron metabolism indicators (FE, TIBC, TS) were included to evaluate the independent predictive effect of ferritin levels on the therapeutic effectiveness, calculate the corrected OR value and 95% CI. The predictive effect was evaluated by the ROC curve. Results: Among patients with chronic hepatitis B (CHB) who completed the PEG IFN alpha-2b course of treatment, the serum ferritin levels of all patients showed an increasing trend at baseline and during the treatment process, and the serum ferritin level in the effective treatment group (HBsAg reduction >= 1 log10 IU/mL or clearance) was significantly higher than that in the ineffective treatment group. It was confirmed by Spearman's rank correlation analysis that the serum ferritin level was positively correlated with the decrease in HBsAg. Further analysis through the multivariate logistic regression model revealed that serum ferritin remained an independent predictor of the antiviral efficacy of

摘要： INTRODUCTION:The residual risk of chronic kidney disease (CKD) progression remains high in clinical trials of kidney protective drugs in patients with diabetic kidney disease (DKD). METHODS:In a prospective study, we assessed whether 16 plasma and 10 urine cytokine levels can inform the residual risk of CKD progression in 93 incident patients with DKD treated by Nephrology according to clinical guidelines. RESULTS:Plasma and urine levels of 12 plasma and 7 urinary cytokines differed between patients with DKD and from healthy controls. Participants were categorized into CKD G1-G2 (preserved GFR) and CKD G3-G5 (GFR <60 ml/min/1.73 m2). After a median of 7.27 years (IQR; 5.34-9.56), 13/40 (32.5%) patients with CKD G1-G2 at baseline had progressed to CKD G3-G5. Progressors had higher plasma IL-22 and TNF-α levels than non-progressors. Plasma IL-22 and TNF-α levels in progressors were similar to those in patients already in CKD G3-G5 at baseline, suggesting that cytokine dysregulation precedes CKD progression. In patients with CKD G1-G2, cut-off points for plasma IL-22 and TNF-α predicted progression with an AUC of 0.76 and 0.77, respectively. Additionally, patients with CKD G1-G2 and plasma TNF-α or IL-22 levels equal to or above the cut-off value had significantly lower eGFR values at the end of follow-up and had more frequently progressed to a very high risk KDIGO category. In cluster analysis, clusters displaying the highest urinary or plasma cytokine levels were associated with worse GFR outcomes. CONCLUSION:Plasma IL-22 and TNF-α may help to identify patients with early DKD with a high residual risk of CKD progression despite treatment.