International Immunopharmacology

Sepsis remains a life-threatening condition with limited therapeutic options targeting immune dysregulation. The CD47-SIRP "dont eat me" signaling axis, well characterized in tumor immune evasion, has not been systematically investigated in the context of sepsis. In this study, we performed a targeted transcriptomic analysis of phagocytosis- and "dont eat me" -related genes using the GSE228541 dataset (14 sepsis patients, 15 healthy controls). We identified 8 significantly differentially expressed genes within the curated gene panel. Key changes included downregulation of CD47 (logFC = -0.88, FDR = 5.6 x 10-4) and marked upregulation of PRTN3 (logFC = 2.68, FDR = 6.1 x 10-4). Gene Ontology (GO) enrichment demonstrated prominent alterations in pathways including negative regulation of phagocytosis (GO:0050765, FDR = 7.6 x 10-22), endocytosis, and inflammatory responses. Co-expression network analysis identified SNX3, DYSF, and PLSCR1 as hub genes within this regulatory module. Immune infiltration analysis showed increased M1 macrophage polarization and neutrophil activation in sepsis. Using LASSO regression, we constructed a 6-gene diagnostic signature (PLSCR1, SNX3, DYSF, PRTN3, CSK, CD47) that discriminated sepsis from controls with good performance (AUC = 0.933 in the test subset). Downregulation of CD47 suggests impaired "self" recognition, which may contribute to aberrant phagocytosis during sepsis. Elevated PRTN3 is consistent with neutrophil activation and extracellular trap formation, linking innate immune activation to tissue injury. This targeted transcriptomic analysis reveals coordinated transcriptional reprogramming of phagocytosis-regulatory genes in sepsis and supports the CD47-SIRP axis as a candidate therapeutic target for further investigation.

Osteoporosis affects millions of women globally. In this study, we applied bioinformatics methods to screen for novel diagnostic biomarkers of osteoporosis in women using the GSE62402 and GSE56814 datasets. PCSK5, ZNF225, and H1FX were used to construct a diagnostic model. ROC, calibration, and decision curve analyses were performed to assess the diagnostic performance on the training (GSE56814) and external (GSE56815) datasets. The expression level of model genes was validated in GEO datasets. Furthermore, five transcription factors (ETS1, NOTCH1, MAZ, ERG, and FLI1) were identified as common upstream regulators of model genes. PCSK5, ZNF225, and H1FX serve as novel diagnostic biomarkers, providing new insights into the pathogenesis of and treatment strategies for osteoporosis in women.

Angiogenesis, the process by which new blood vessels form from existing vasculature, is fundamental to tissue repair and regeneration but also underlies pathological conditions such as cancer progression. Targeting angiogenesis has thus become a promising approach for developing novel cancer therapeutics. While various phytochemicals have demonstrated anti-angiogenic effects, the role of 2-5(H)-Furanone, a naturally occurring lactone found in various plants and marine sources with diverse biological activities, remains insufficiently explored. In this study, we systematically evaluate the anti-angiogenic potential of 2-5(H)-Furanone using Human Umbilical Vein Endothelial Cells (HUVECs) as an in vitro model and zebrafish embryos as an in vivo model. Experimental findings demonstrated that treatment of HUVECs with increasing concentrations of 2-5(H)-Furanone led to significant, dose-dependent reductions in proliferation, invasion, migration, and tube formation. Analyses of gene expression revealed marked downregulation of key pro-angiogenic mediators, VEGF, and HIF-1. Complementing these in vitro results, in vivo studies in zebrafish embryos showed robust, dose-dependent inhibition of intersegmental vessel (ISV) formation, accompanied by suppression of critical angiogenesis-related genes. Molecular docking further supported these observations by indicating stable binding of 2-5(H)-Furanone to major angiogenic targets, including VEGFR2, MMP2, HIF-1, and PIK3CA. Collectively, our data demonstrate that 2-5(H)-Furanone potently inhibits angiogenesis, as evidenced in both HUVEC and zebrafish models, through functional and molecular mechanisms. These findings support the further development of 2-5(H)-Furanone as a promising anti-angiogenic therapy candidate.

Class B scavenger receptors BI (SR-BI) and BII (SR-BII) internalize lipoproteins but also bind and internalize bacteria. Their roles in sepsis are unknown. We overexpressed human SR-BI and BII in the liver and kidney as well as bone marrow-derived macrophages, and then performed cecal ligation and puncture (CLP) surgery. SR-BI and BII transgenic mice had significantly worse survival compared to WT mice. 24 h after CLP, liver injury markers and histological damage were prominent in both SR-BI and BII transgenic mice, whereas kidney damage was similar. Systemic inflammatory cytokines were markedly increased in SR-BI and BII transgenic mice; parallel increases were seen in liver mRNA expression, not in the kidney. The highest degree of neutrophil infiltration was observed in the liver of SR-BI. Human SR-BI and BII dramatically decreased bacterial accumulation in the liver. Green fluorescence protein-labeled E. coli were efficiently phagocytosed in hepatic macrophages of SR-BI and BII transgenic mice; phagocytosis was more prominent in SR-BII transgenic mice. Finally, human SR-BI overexpression reduced systemic HDL-C level, eliminated adrenal cortex lipid droplets, and dampened the systemic increase of corticosterone after CLP. Supplementation with glucocorticoid and mineralocorticoid improved survival in SR-BI, but not SR-BII, transgenic mice after CLP. In summary, our findings suggest human SR-BI and BII overexpression contributes to higher mortality after CLP by excessive inflammatory response due to adrenal insufficiency (SR-BI) or hyperactive phagocytosis (SR-BII) in the liver.

Frailty is a prevalent geriatric syndrome, and the shortage of objective biomarkers restricts its early diagnosis and intervention. This study aimed to identify robust molecular signatures and diagnostic markers for frailty using bioinformatics analyses of multiple independent datasets. Two transcriptome datasets (GSE144304, n=80; GSE287726, n=70) were obtained from the GEO database. We performed differential gene expression analysis, GO, KEGG and GSEA enrichment, and machine learning (70% training / 30% validation) to screen and validate core biomarkers. Numerous shared differentially expressed genes were identified. Vitamin D metabolism, ABC transporter, and inflammatory/immune pathways were consistently enriched and confirmed by GSEA. Machine learning models based on these signatures showed favorable diagnostic performance. Our study demonstrates that vitamin D metabolic disorders and chronic inflammation are core molecular features of frailty. The identified biomarkers provide new strategies for basic research, early clinical diagnosis, and therapeutic target development for frailty.

Mesenchymal stem cells (MSCs) cultured in hypoxic conditions have been suggested to have more therapeutic efficacy than those cultured under normoxic conditions, and there is growing interest in using hypoxic MSCs for clinical treatment, particularly human umbilical cord (hUC)-MSCs. We investigated how hUC-MSCs and human bone marrow (hBM)-MSCs change from normoxia to hypoxia (1% O2) for 2 weeks of culture. In the growth speed and population doubling time, hUC-MSCs cultured under hypoxia exhibited a significantly higher proliferation rate beyond cancerous cells, such as human glioblastoma and breast cancer cells, while hBM-MSCs did not show a significant difference between normoxia and hypoxia, and were statistically slower than these cancerous cells. Notably, hypoxic hUC-MSCs showed upregulation of genes related to metabolic reprogramming (cholesterol biosynthesis and fatty acid metabolism pathways) and cancer stem cell-like phenotype (factors related to Wnt and Hedgehog signaling pathways, cell proliferation drivers, and apoptosis-resistance), and lesser migration and homing to the traumatic brain injury than normoxic hUC-MSCs after intravenous injection. Thus, whether hUC-MSCs cultured under hypoxia offer clinical benefits and use are safe, given their extremely accelerated proliferation rate and partial cancer stem cell-like traits, requires comprehensive and careful investigation.

Intervertebral disc degeneration, a leading cause of low back pain with incompletely elucidated molecular mechanisms, was studied via integrated in vivo/vitro approaches. This study first reveals that lactic acid accelerates intervertebral disc degeneration by inducing cartilage endplate stem cells senescence and DNA damage, thereby activating the P16/P21/P53-centered senescence pathway. In a rat tail vertebra puncture-induced intervertebral disc degeneration model, degenerated discs exhibited increased lactic acid levels, narrowed intervertebral spaces, and disrupted nucleus pulposus structure (P<0.05). In vitro, 0/2/6/10 mM lactic acid dose-dependently suppressed cartilage endplate stem cells viability (10 mM group: 15.7% of the control), elevated intracellular reactive oxygen species (ROS, 2.8-fold relative to the control), induced G0 cell cycle arrest (10 mM group: 85.63%), reduced EdU-positive cells (8.62%), and increased {beta}-galactosidase-positive cells (10 mM group: 33.06%) and {gamma}-H2AX foci (all P<0.01).Molecularly, lactic acid significantly upregulated P16 (2.1-fold), P21 (3.1-fold), P53 (2.4-fold), and {gamma}-H2AX (1.8-fold). In vivo intervertebral disc injection confirmed a positive correlation between lactic acid concentration and intervertebral disc degeneration severity. This study clarifies lactic acids role in intervertebral disc degeneration via the "oxidative stress-cell cycle arrest-cellular senescence" axis, advancing understanding of intervertebral disc degeneration pathogenesis and providing a basis for targeted therapies against lactic acid metabolism.

BackgroundWhen the skin wound defect is too large, it is difficult for the body to heal itself, and medical treatment is often needed. How to shorten the healing cycle and reduce the incidence of infection is a difficult problem faced by clinicians. Panax notoginseng(PN), a traditional Chinese medicine, can promote the absorption of inflammatory exudates, granulation tissue formation and epidermal proliferation, effectively inhibit the inflammatory reaction of wounds and promote the healing of skin wounds, but its molecular mechanism has not been fully clarified so far. Based on network pharmacology and animal experiments, this study explored the target and molecular mechanism of PN in the treatment of skin wound. MethodsThe active components and potential targets of PN were obtained from the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP) and UniProt database, and the skin wound-related targets were obtained from the GeneCards database. The intersecting targets were filtered using Venny 2.1.0. The intersecting targets were imported into the STRING database to construct a protein-protein interaction (PPI) network. Cluster analysis was performed using the MCODE and CytoHubba plugins in Cytoscape 3.8.2 to obtain core functional network modules and the top 10 key target genes. The intersecting targets were subjected to KEGG and GO enrichment analysis using the DAVID 6.8 database (https://david.ncifcrf.gov/). The component-target-pathway network of PN in the treatment of skin wounds was constructed using Cytoscape 3.8.2 software. In the experimental verification phase, 48 Sprague-Dawley (SD) rats were randomly divided into a control group and a PN group, with 24 rats in each group. A full-thickness skin excision was performed to establish a wound model. The PN group received intraperitoneal injections of the drug, while the control group received an equivalent amount of saline. Wound area measurements were taken on days 1, 4, and 7 after model establishment. Histopathological changes in the injured area and the expression and localization of TNF-, IL-6, and IL-10 were observed through hematoxylin and eosin (HE) staining and immunohistochemical staining. Relative expression levels of the three factors were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). ResultsThis study identified 8 major active components, 156 targets, and 115 signaling pathways involved in the treatment of skin wounds in rats using PN. The top 10 core target genes included TNF, IL-6, and IL-10, primarily enriched in signaling pathways such as NF-{kappa}B, MAPK, and JAK-STAT. Animal experiments revealed that at 4 and 7 days post-injury, the wound area in the PN group was significantly smaller than that in the control group (P<0.05). HE staining showed reduced infiltration of neutrophils and inflammatory cells in the injury area at 7 days in the PN group, accompanied by more pronounced fibroblast proliferation and collagen secretion. Molecular detection indicated that TNF-, IL-6, and IL-10 positive reactants were mainly distributed in the cytoplasm and matrix of epidermal cells, inflammatory cells, and fibroblasts in the skin. qRT-PCR and ELISA results showed that TNF- expression in the PN group was significantly lower than that in the control group at 4 and 7 days (P<0.01). IL-6 expression was lower than that in the control group at all time points, peaking at 4 days and then decreasing (P<0.01). IL-10 expression was significantly lower than that in the control group at 1 and 7 days (P<0.01). ConclusionPN exhibits characteristics such as multi-component, multi-target, multi-pathway synergy, and multiple regulatory pathways in the treatment of skin wounds. It can reshape the dynamic balance of cytokine networks, including TNF-, IL-6, and IL-10, optimize the temporal progression of "inflammation initiation - repair transition - tissue remodeling", and provide a therapeutic effect of "efficient debridement - orderly repair - low scar risk" for skin wounds. It is one of the ideal natural drugs for regulating skin wound healing.

Mucopolysaccharidosis III (MPS III or Sanfilippo disease) is a spectrum of 4 genetic disorders (MPS IIIA-D), caused by defects in the genes SGSH, NAGLU, HGSNAT and GNS encoding enzymes involved in degradation of heparan sulfate (HS). HS accumulates in brain tissues and causes neuronal dysfunction and neurodegeneration leading to neuropsychiatric problems, developmental delays, childhood dementia, blindness and death during the second decade of life. Previously, we demonstrated that pathophysiological mechanisms, underlying MPS IIIC in mouse models, involves functional pathological changes, affecting synaptogenesis and synaptic transmission and leading to learning and memory deficits. These results suggested that a treatment for MPS III could be developed by using compounds inducing synaptogenesis. In the current study, we tested the efficacy of a synthetic peptide ACTH(4-7)PGP, an analog of adrenocorticotropic hormone fragment, previously used as a neuroprotective and anti-inflammatory medication for treatment of acute neurological conditions, including stroke. We show that intranasal administration of ACTH(4-7)PGP restores defective synaptic transmission in CA1 pyramidal neurons of MPS IIIA and MPS IIIC mouse models and rescues the decrease in synaptic proteins in cultured MPS IIIC mouse hippocampal neurons and iPSC-derived neurons of human MPS IIIA, MPS IIIB and MPS IIIC patients. Furthermore, daily intranasal administration of ACTH(4-7)PGP to MPS IIIC and MPS IIIA mice reduces hyperactivity and rescues defects in working and spatial memory, delays progression of CNS pathology including neuroinflammation and axonal demyelination, and increases the lifespan. Together with the absence of any adverse reactions to ACTH(4-7)PGP in the MPS III and WT mice, our results justify testing the drugs efficacy in clinical settings.

Sepsis is defined as a dysregulated response to infection, leading to life-threatening organ dysfunction that particularly affects parenchymal organs. Clinical studies remain inconclusive regarding the impact of biological sex on sepsis, and preclinical studies are predominantly performed in male animals. We examined early (8 h) septic responses in male and female mice using a fecal-induced peritonitis (FIP) model. Blood biochemical parameters, body temperature, and murine sepsis scores provided evidence of a septic response in animals randomized to FIP compared to controls, but showed no physiological differences between male and female mice. Transcriptomic analysis of the liver, kidney, and lung showed consistent inflammatory activation in response to sepsis as compared to controls. Notably, in the kidney and lung, female mice exhibited stronger immune activation and a heightened inflammatory response compared to males. Thus, biological sex differences in the septic response can be detected in early acute sepsis without apparent physiological differences.

BACKGROUNDMyocardial ischemia/reperfusion injury (I/RI) represents a serious clinical complication in patients after acute myocardial infarction. Ubiquitin-activating enzyme 1 (UBA1) catalyzes the initial step of ubiquitination and plays a fundamental role in regulating protein homeostasis and related diseases. This study aims to elucidate the functional contribution of UBA1 to the pathogenesis of myocardial I/RI and to uncover its underlying mechanisms. METHODSSingle-cell RNA sequencing was employed to characterize UBA1 expression in human ischemic heart tissues. Myocardial I/R injury was examined in myocardial-specific UBA1 knockout (UBA1cko) mice, UBA1-overexpressing mice (rAAV9-UBA1), and corresponding controls. Neonatal rat cardiomyocytes underwent hypoxia/reoxygenation in vitro. Cardiac function and infarction were evaluated by echocardiography and pathological staining. Protein-protein interactions were analyzed via immunoprecipitation combined with mass spectrometry. The endoplasmic reticulum-mitochondrial contact sites (ERMCSs) and mitochondrial ultrastructure were evaluated through transmission electron microscopy and confocal imaging. RESULTSUBA1 expression was significantly downregulated in human and murine ischemic myocardium, especially in cardiomyocytes. UBA1cko mice exhibited aggravated I/RI with greater infarct size, impaired function, apoptosis, elevated intracellular Ca2+ levels, mitochondrial dysfunction, and ER stress, whereas UBA1 overexpression conferred cardioprotective effects. Mechanistically, UBA1 directly bound to and ubiquitinated Pdzd8, a key ERMCS-tethering protein, thereby promoting its degradation, which inhibited ERMCS formation and improved mitochondrial dysfunction and ER stress. Moreover, knockdown of Pdzd8 via rAAV9-siRNA effectively mitigated UBA1 knockout-induced myocardial damage. Additionally, administration of auranofin (AF), a U.S. Food and Drug Administration-approved drug for treating rheumatoid arthritis, markedly alleviated myocardial I/RI via activating UBA1 in vivo and in vitro. CONCLUSIONSUBA1 confers protection against myocardial I/RI by limiting ERMCS formation through Pdzd8 ubiquitination. Activating UBA1 or targeting Pdzd8 as a potential therapeutic strategy for the treatment of ischemic heart disease. GRAPHIC ABSTRACTA graphic abstract is available for this article. Clinical PerspectiveO_ST_ABSWhat Is New?C_ST_ABSO_LIUBA1 expression is downregulated in human and murine ischemic myocardium, especially in cardiomyocytes. C_LIO_LICardiac deletion of UBA1 significantly exacerbates myocardial ischemia/reperfusion injury (I/RI), whereas cardiac UBA1 overexpression confers a marked protective effect. C_LIO_LIUBA1 interacts with Pdzd8 (PDZ domain containing 8) and facilitates its ubiquitination and subsequent degradation, which then reduces endoplasmic reticulum-mitochondria contact sites (ERMCSs) and ameliorates mitochondrial dysfunction and ER stress, protecting myocardial I/RI. C_LIO_LIPharmacological activation of UBA1 with the FDA-approved drug auranofin attenuates myocardial I/R injury and improves heart dysfunction. C_LI What Are the Clinical Implications?O_LIUBA1 represents a new therapeutic target for myocardial I/RI. C_LIO_LIActivating UBA1 or targeting Pdzd8 may offer a promising therapeutic strategy for mitigating myocardial I/RI and heart failure, underscoring its potential for clinical translation. C_LI

Social stress is a risk factor for psychiatric disorders and also influences immune function. While it is known that acute social stress impacts the number of immune cells in circulation, the temporal dynamics of stress induced immune-related transcriptional changes in human blood remain unclear. To investigate changes in gene expression, we exposed 26 adults to the Trier Social Stress Test (TSST), and collected blood at baseline, as well as 5, 30, 60 and 90 min after stress. Whole-blood gene expression was profiled using a 5 targeted RNA-sequencing method (STRT). Differential expression was analyzed using linear and cubic models. We observed a total of 54 differentially expressed genes following stress. Fast responses, with a transient peak immediately following stress, were enriched for cytotoxic T cell, NK cell and dendritic cell functions (e.g., GZMB, GNLY, CCL4 and GZMA) and paralleled lymphocyte count changes. In contrast, gradual, linear responses without any evident peak were enriched for neutrophil related genes (e.g., FPR2, PLAUR, CXCR2, AQP9, and QPCT) and did not mirror neutrophil counts, indicating cell intrinsic transcriptional changes. From pathway and transcription factor enrichment analyses, IL-12 family mediated signaling is inferred as a central mechanism linking stress to immune gene regulation. Our results show that acute psychosocial stress induces both fast and slower changes in gene expression in different immune cell populations. The involvement of the IL-12-STAT4 axis and genes such as PLAUR and FPR2 suggests molecular mechanisms through which stress-related immune activation may contribute to vulnerability for anxiety and depressive disorders.

Background The high mortality of septic shock demands novel adjunctive therapies. Shenfu Injection (SFI), a traditional Chinese medicine, shows potential but its mechanism remains unclear. Method s We conducted an open-label, randomized trial in 80 patients with septic shock. Patients received standard care with or without adjunctive SFI for 7 days. The primary outcome was 28-day mortality. Key secondary outcomes included inflammatory markers, lactate clearance, and vasopressor duration. Concurrently, network pharmacology analyzed SFIs bioactive components, predicted targets, and enriched pathways, with validation by molecular docking. Results The 28-day mortality was significantly lower in the SFI group (20.0% vs. 42.5%, P=0.030). SFI accelerated clinical improvement, evidenced by greater reductions in IL-6 and procalcitonin, higher 6-hour lactate clearance (35.2% vs. 18.5%, P<0.001), shorter vasopressor duration (48 vs. 72 hours, P<0.001), and more rapid SOFA score decline. Network pharmacology identified 145 SFI-septic shock common targets, with IL-6, SRC, and MAPK3 as central hubs. Pathway analysis revealed significant enrichment in TNF, PI3K-Akt, and IL-17 signaling pathways. Molecular docking confirmed strong binding of key SFI components (e.g., Ginsenoside Rh2) to core targets like IL-6. Conclusion s Adjunctive Shenfu Injection reduces mortality and improves clinical recovery in septic shock, potentially through a multi-target mechanism involving modulation of inflammatory and cellular signaling pathways. This integrative study provides both clinical evidence and a mechanistic framework supporting SFI's use. Clinical Trial Registration: Chinese Clinical Trial Registry, ChiCTR1800020435.

This study investigates the therapeutic potential of secretomes derived from Adipose-derived Mesenchymal Stem Cells (ADMSC-CM) and Limbal-derived Mesenchymal Stem Cells (LMSC-CM) against oxidative stress-induced damage in Retinal Pigment Epithelium (RPE-1) cells. RPE dysfunction, often triggered by oxidative stress, is a hallmark of various retinal degenerations. Here, we induced RPE-1 injury using H2O2 and evaluated the restorative effects of both MSC-conditioned media (CM). Our results demonstrated that both ADMSC-CM and LMSC-CM significantly enhanced cell viability and successfully reversed H2O2-induced G2/M phase cell cycle arrest. While oxidative stress triggered a pro-inflammatory response characterized by elevated IL-1{beta}, IL-6, and IL-10 expression, MSC-CM treatment, particularly ADMSC-CM, effectively modulated these levels and suppressed the p38 MAPK signaling pathway. Furthermore, MSC-CM reduced the Bax/Bcl-2 ratio, indicating an anti-apoptotic effect, and appeared to stabilize autophagic flux. To investigate the impact of oxidative-stress induced alterations in retinal pigment epithelial cells on angiogenesis, the effects of RPE-derived secreted factors on endothelial cell function were evaluated. Crucially, in terms of safety and secondary complications, neither secretome exhibited pro-angiogenic tendencies; instead, they significantly inhibited HUVEC migration and invasion compared to the H2O2 damaged group. These findings suggest that both ADMSC and LMSC secretomes provide a potent multi-targeted therapeutic effect, making them promising candidates for cell-free therapies in retinal diseases.

Regulator of G-protein signaling 10 (RGS10) has been shown to regulate multiple inflammatory pathways relevant to disease pathogenesis. Of particular importance is the ability of RGS10 to negatively regulate the NFkB pathway, a prominent pro-inflammatory pathway implicated in multiple inflammatory disease phenotypes. However, the exact mechanism by which RGS10 regulates NFkB is unknown. Considering that RGS10 translocates into the nucleus upon stimulation, we hypothesize that RGS10 may regulate NFKB through transcription. To determine whether RGS10 mediates NFkB transcription, we stimulated RGS10 KO and B6 peritoneal macrophages and collected cell lysate over 24 hours to assess transcript levels of NFkB and related proinflammatory cytokines. Here we found that RGS10 differentially regulates the transcription of N{kappa}KB subunits and NF{kappa}B-dependent cytokines. Further studies are warranted to understand the potential role of RGS10 in transcriptional regulation of inflammatory states.

Inter-alpha Inhibitor Proteins (IAIP) are serine protease inhibitors and a reliable inflammatory biomarker. We have demonstrated that IAIP levels decrease during sepsis in humans and animal models. Currently, enzyme-linked immunosorbent assay (ELISA) is the standard procedure to measure IAIP levels. We developed a lateral flow immunoassay (LFIA) that allows rapid, point of care detection. We compared IAIP levels in infants with sepsis and/or necrotizing enterocolitis (NEC) by ELISA and LFIA in a multi-center, cross-sectional study. Blood samples were collected from 47 infants with sepsis, 31 sepsis case controls, 52 gestational age (GA)-matched controls and 10 infants with culture-negative sepsis ("clinical sepsis"). We also collected samples from 17 infants with NEC, 7 NEC case controls and 15 GA controls. IAIP levels at presentation of acute events and over the next 72 hours were significantly reduced in infants with sepsis, NEC and culture negative sepsis when compared to controls. IAIP levels did not differ in patients with sepsis or culture negative sepsis. IAIP levels measured by ELISA and LFIA were highly correlated (R2 = 0.9326) and both showed reliable detection of neonatal sepsis, NEC and culture negative sepsis. IAIP levels were 80.0% sensitive and 92.3% specific using LFIA for the detection of neonatal sepsis. For detection of NEC, IAIP levels were 84.6% sensitive and 86.7% specific.

The IRE1-XBP1 axis is the most conserved of three major unfolded protein response (UPR) branches that are triggered by the endoplasmic reticulum (ER) stress. Although the transcription factor XBP1 is involved in the development and function of several hematopoietic lineages, the role of XBP1 in the activation of mast cells (MCs) that play key role in allergic response remains largely unknown. Because we have identified salicylaldehyde (SA), which inhibits IRE1 nuclease activity that is essential for production of XBP1, as an inhibitor of MC activation in our previous screening, we investigated the effects of additional IRE1 inhibitors, 3-methyl-6-bromo-salichylaldehyde (MBSA) and KIRA6, targeting nuclease domain and kinase domain, respectively, on MC activation. MBSA and KIRA6 suppressed IgE-dependent degranulation and cytokine release of bone marrow-derived MCs (BMMCs), whereas these inhibitors did not suppress the Ca2+ ionophore- or compound48/80-induced degranulation. Treatment with inhibitors against two other branches of UPR, the PERK and the ATF6 pathways, did not affect IgE-induced activation of BMMCs. Intraperitoneal administration of MBSA or KIRA6 significantly suppressed IgE-induced passive anaphylaxis in mice. Furthermore, to evaluate the effect of XBP1, siRNA-mediated knockdown was performed. It was confirmed that Xbp1 siRNA introduction reduced IgE-dependent degranulation of BMMCs in parallel with the knockdown level of Xbp1 mRNA. Taken together, the IRE1-XBP1 axis plays a significant role in IgE-dependent and MC-mediated allergic response, which is considered to be therapeutic target of allergic diseases.

Objective. To evaluate the diagnostic and prognostic significance of C reactive protein (CRP) level dynamics within the first five days after surgery for the early detection of surgical site infections (SSI) and to identify independent risk factors, taking into account regional specifics of surgical management (types of surgeries, duration of procedures), as well as the local hospital microbial landscape. Materials and Methods. A single-center retrospective cohort analysis of data from 127 patients who underwent surgical procedures between 2022 and 2024 was conducted. CRP levels on postoperative days 1, 3, and 5 were assessed, and delta values were calculated. Descriptive statistics, ROC analysis, and multivariate logistic regression were used to identify predictors of SSI. Results. Patients with SSI lacked the physiological decrease in CRP levels by day 5. The most informative indicator was the CRP level on day 3: a threshold of >106 mg/L was associated with a high risk of SSI (AUC=0.76; sensitivity 85%, specificity 63%). Independent predictors of SSI included surgery duration (OR=1.015 per 1 min; p<0.001) and the increase in CRP between days 3 and 5 (delta CRP3-5: OR=1.027; p=0.023). A combined model (clinical parameters + CRP) demonstrated the highest predictive ability (AUC=0.79). Conclusion. Monitoring CRP dynamics, particularly on days 3 and 5, is a highly informative and accessible method for the early diagnosis of SSI. A CRP threshold of >100 mg/L on day 3 and its subsequent increase should serve as a trigger for in-depth diagnostic investigation and rationalization of antimicrobial therapy. Keywords: C reactive protein, postoperative complications, surgical site infection, antibiotic therapy, predictive factors, diagnosis

ObjectiveTo explore the application value of dual-staining for specific AT sequence binding protein 2 (SATB2) immunohistochemistry and elastic lamina in detecting elastic lamina invasion (ELI) in pT3 colon cancer, and to assess its association with clinicopathological characteristics, staging, and prognosis. MethodsThis retrospective cohort study enrolled 176 pT3 colon cancer patients who underwent radical resection at Affiliated Jinhua Hospital Zhejiang University School of Medicine. The deepest tumor-infiltrated paraffin blocks were collected for SATB2 immunohistochemistry and elastin dual-staining. Correlations between ELI status and clinicopathological characteristics and prognosis were analyzed. Survival data of 74 pT4a stage patients were collected for comparative analysis. ResultsELI (+) was positively associated with high tumor budding grade, vascular invasion, lymph node metastasis, and reduced tumor infiltrating lymphocytes (TILs) (all P < 0.001). No correlations were observed with age, gender, tumor location, histological subtype, tumor grade, or perineural invasion (all P > 0.05). The ELI (+) group exhibited significantly shorter disease-free survival (DFS) and overall survival (OS) compared to ELI (-) group (P < 0.05). Additionally, the ELI (+) group demonstrated inferior OS than the pT4a group, though DFS did not differ significantly. ConclusionDual-staining of SATB2 immunohistochemistry and elastic lamina provides a reproducible and objective method for assessing ELI. ELI correlates with key clinicopathological features and functions as an independent adverse prognostic indicator in pT3 colon cancer.

BackgroundInterferon-gamma (IFN-{gamma}) is the primary effector cytokine of adaptive anti-tumor immunity, yet it paradoxically induces a potent immunosuppressive tumor microenvironment (TME). The full mechanistic scope of this paradox in head and neck squamous cell carcinoma (HNSC) has not been characterized at the transcriptomic scale. MethodsUsing TCGA HNSC RNA-seq data (n = 522), we applied an integrated computational pipeline: Spearman correlation analysis, principal component analysis (PCA), UMAP, K-means clustering (k = 4), Random Forest regression, deep neural networks, permutation importance, JAK-STAT cascade mapping, and DNN-based transcriptome-wide mediation analysis across 57 IFN-{gamma} pathway and 78 immunosuppressive genes. ResultsIFN-{gamma} pathway activity was universally and positively correlated with six immunosuppressive axes, including checkpoints (CD274; LAG3; IDO1), Tregs, myeloid suppression, and tryptophan catabolism. K-means clustering identified four immunologically distinct tumor subgroups. DNN models predicted suppressive TME. Permutation importance identified IRF8 as the dominant mediator linking IFN-{gamma} signaling to immunosuppression. DNN mediation analysis identified PDCD1LG2 (PD-L2) as the strongest intermediary between IFNG and PD-L1 regulation, followed by JAK2 and GBP5. ConclusionsIFN-{gamma} orchestrates coordinated immunosuppression in HNSC through JAK-STAT-IRF8 signaling. PDCD1LG2 and JAK2 are actionable mediators of this paradox, supporting combination strategies co-targeting IFN-{gamma}-induced checkpoint induction and direct checkpoint blockade in HNSC immunotherapy. GRAPHICAL ABSTRACT