Journal of Cellular and Molecular Medicine

Emerging evidence indicates an intricate relationship between the SARS-CoV-2 infection and Multi-Organ Dysfunctions (MODs). Here, we have investigated the role of the Secretome of the SARS-CoV-2 infected pancreas and mechanistically linked it with the multi-organ dysfunction using the scRNA-seq analysis. We found that acinar-specific PRSS2, REG3A, REG1A, SPINK1, and ductal-specific SPP1, MMP7 genes are upregulated in alpha, beta, delta, and mesenchyme cells. Using extensive documented experimental evidence, we validated the association of upregulated pancreatic Secretome with coagulation cascade, complement activation, renin angiotensinogen system dysregulation, endothelial cell injury and thrombosis, immune system dysregulation, and fibrosis. Our finding suggests the influence of upregulated Secretome on multi-organ systems such as Nervous, Cardiovascular, Immune, Digestive, and Urogenital systems. In addition, we report that the secretory proteins IL1B, AGT, ALB, SPP1, CRP, SERPINA1, C3, TFRC, TNFSF10, and MIF are associated with diverse diseases. Thus, suggest the role of the pancreatic Secretome in SARS-CoV-2 associated MODs.

BackgroundSepsis-induced cardiomyopathy (SIC) represents a critical complication of sepsis, characterized by reversible myocardial dysfunction and alterations. Despite extensive research, the molecular mechanisms underlying SIC remain poorly understood. MethodsUtilizing bioinformatics analysis of RNA-seq and scRNA-seq data from the GEO database, we identified key immune cell populations and molecular markers associated with SIC. Our in vitro and in vivo studies focused on the roles of ITGAM and ICAM-1 in macrophage recruitment and transformation as well as the impact of these changes on cardiac function. ResultsBioinformatics analysis revealed significant alterations in gene expression and immune cell composition in cardiac tissue during SIC, with macrophages being the predominant immune cell type. ITGAM was identified as crucial molecule in this process. In vitro experiments demonstrated an upregulation of ITGAM in macrophages and ICAM-1 in endothelial cells following LPS stimulation, indicating their roles in immune cell recruitment and interaction. Furthermore, the use of ITGAM-neutralizing antibodies led to reduced macrophages infiltration and initially improved cardiac function in SIC mice, but resulted in increased mortality in later stages. These findings highlight the dual role of ITGAM in SIC, facilitating early-stage inflammation and later-stage cardiac recovery. ConclusionThis study elucidates the complex dynamics of immune cells in SIC, with a particular emphasis on the role of ITGAM in macrophage modulation. The findings provide new insights into the reversible nature of myocardial dysfunction in SIC and underscore the importance of targeted therapeutic strategies for effective sepsis management. HighlightsIdentifies ITGAM as a key modulator in macrophage dynamics during sepsis-induced cardiomyopathy (SIC). Elucidates the impact of ITGAM on cardiac function in SIC. Reveals new insights into the immune-cellular mechanisms in SIC pathology.

ObjectiveInflammation is a critical process during the progressive development and complication of abdominal aortic aneurysm. The co-stimulatory dyad CD40-CD40L is a major driver of inflammation and modulates immune responses. This study evaluates the potential of a small molecule inhibitor, which blocks the interaction between CD40 and tumor necrosis factor (TNF) receptor-associated factor (TRAF)-6, referred to as TRAF-STOP, in the early and later phase during AAA progression. Methods and resultsAAAs were induced in C57BL/6J mice by infrarenal aortic porcine pancreatic elastase infusion for 7, 14 or 28 days. Inhibition of CD40 signaling by TRAF-STOP resulted in less severe AAA formation and reduced the incidence of AAA development. TRAF-STOP treatment attenuated aortic structural remodeling, characterized by a reduced elastic fiber degradation, lowered expression of matrix metalloproteinase (MMP)-2 and MMP9, as well as preserved collagen type IV content in aneurysmal tissue. Furthermore, this is accompanied by the reduction of key pro-inflammatory genes such as TNF. ConclusionPharmacological inhibition of CD40-TRAF6 signaling protects from adverse aortic structural remodeling during the early phase of AAA progression representing a translational strategy to limit progression of human AAA disease.

Background and aimsNeutrophil extracellular traps (NETs), fibrin, and the von Willebrand factor (VWF) are present in acute coronary thrombotic plaques. However, the presence and extent of NETs, fibrin, and VWF in early-to-unstable coronary atherosclerotic lesions remain unclear. This study aims to determine the presence and extent of neutrophils, NETs, fibrin, and VWF formation during human coronary atherogenesis. MethodsCoronary sections from autopsy patients with non-cardiac death (n=5, 37 sections) and ischemic heart diseases (n= 11, 65 sections) were classified by atherosclerosis classification: diffuse intimal thickening, pathological intimal thickening, fibrous cap atheroma, fibrocalcified plaque, thin-cap fibroatheroma, ruptured plaque, and intraplaque hemorrhage. We immunohistochemically assessed the expression of CD66b (neutrophils), citrullinated histone H3 (Cit-H3, a marker of NETs), fibrin, and VWF. ResultsNeutrophil and Cit-H3 expression were rarely observed, except in thin-cap fibroatheromas and ruptured plaques. Fibrin deposition was observed in pathological intimal thickening and necrotic cores of atheromas, and it was abundant in thin-cap fibroatheroma and ruptured plaques. VWF deposition was observed in the necrotic core of the atheromas and was abundant in ruptured plaques. In non-ruptured plaques, the immunopositive areas for Cit-H3 and fibrin were larger in hemorrhagic plaques than in non-hemorrhagic plaques. ConclusionsThese results suggest that NET formation is rare in stable coronary plaques, and fibrin formation begins in stable lesions and increases with plaque destabilization in the coronary artery. Intraplaque hemorrhage may promote NET formation and fibrin deposition in non-ruptured plaques. The abundance of neutrophilic infiltrates and VWF deposition may be a response to plaque rupture.

During atherosclerosis, smooth muscle cells (SMCs) migrate and accumulate in the intima, where they switch from a contractile to a synthetic phenotype. This process is associated with decreased expression or loss of contractile proteins, such as -smooth muscle actin (-SMA) or smooth muscle myosin heavy chains (SMMHCs). We previously demonstrated that S100A4, a small calcium-binding protein, which exhibits intra- and extracellular functions, is a marker of the synthetic SMC phenotype. We have recently shown that neutralization of extracellular S100A4 in an ApoE knockout (KO) mouse model with established atherosclerotic lesions decreased the overall atherosclerotic burden. To explore the role of S100A4 in the accumulation of SMCs in the intima, we induced intimal thickening (IT) formation in full S100A4 knockout (KO) and wild type (WT) mice by completely ligating the left common carotid artery. With this model, we generated SMC-rich lesions. The deletion of S100A4 did not influence the size of the IT, neither its composition, as assessed by the expression of -SMA and SMMHCs in S100A4 KO animals compared with WT animals, 4 weeks after ligation. Using primary cells isolated from both strains, we demonstrated that S100A4 KO SMCs were less prone to migrate than WT SMCs but they did not differ in their proliferative capacity. Our results indicate that S100A4 plays a role in SMC motility in vitro but its deletion does not influence IT formation in the mouse carotid artery ligation model.

Recurrent pregnancy loss (RPL) affects 1-2% of women trying to conceive, yet in many cases the causes remain unclear. Endometrial function is central to the establishment and maintenance of pregnancy, and endometrial dysfunction may underlie RPL. A greater understanding of the endometrial cell populations and their interactions in women with and without RPL may identify markers of endometrial receptivity and the likelihood of pregnancy success. Single cell RNA sequencing was performed on RPL (n = 3) and control (n = 4) endometrial biopsies collected at days 21-24 of the menstrual cycle, the window of implantation. 10,022 cells were clustered and nine major individual cell types were characterised. Further analysis identified six distinct endometrial stromal cell (EnSCs) and three natural killer (NK) cell sub-populations. In RPL, there were changes in the abundance of specific endometrial stromal and NK cell subpopulations with associated differences in cellular communication between the cell types related to the Wnt pathway and angiogenesis. This is consistent with NK cell signalling orchestrating the difference in abundance of stromal cells and regulating processes needed for successful implantation. These changes in RPL endometrium provide further evidence for an endometrial cause of RPL and identify specific mechanisms for future study.

BackgroundAngiotensin-converting enzyme-2 (ACE2) overexpression improves left ventricular remodeling and function in diabetic cardiomyopathy; however, the effect of ACE2-overexpressed mesenchymal stem cells (MSCs) on myocardial infarction (MI) remains unexplored. This study aimed to investigate the effect of ACE2-overexpression on the function of MSCs and the therapeutic efficacy of MSCs for MI. MethodsMSCs were transfected with Ace2 gene using lentivirus, and then transplanted into the border zone of ischemic heart. The renin-angiotensin system (RAS) expression, nitric oxide synthase (NOS) expression, paracrine factors, anti-hypoxia ability, tube formation of MSCs, and heart function were determined. ResultsMSCs expressed little ACE2. ACE2-overexpression decreased the expression of AT1 and VEGF apparently, up-regulated the paracrine of HGF, and increased the synthesis of Angiotensin 1-7 in vitro. ACE2-overexpressed MSCs showed a cytoprotective effect on cardiomyocyte, and an interesting tube formation ability, decreased the heart fibrosis and infarct size, and improved the heart function. ConclusionTherapies employing MSCs with ACE2 overexpression may represent an effective treatment for improving the myocardium microenvironment and the cardiac function after MI.

BackgroundCalcific aortic valve disease (CAVD) is the most common valvular heart disease in developed countries, yet no pharmacological therapy is available to slow or halt its progression. CAVD is driven by progressive calcification of aortic valve leaflets, in which myeloid cells play a central role. While macrophages have been implicated in CAVD pathogenesis, the contribution of their precursors, monocytes, remains poorly understood. We hypothesized that circulating monocytes acquire a pro-calcific and pro-inflammatory phenotype contributing to valve remodelling and CAVD progression. MethodsWe profiled circulating CD14+ monocytes from healthy volunteers (Vol), patients with CAVD, and without CAVD (NCAVD). Peripheral blood mononuclear cells (PBMCs) were isolated, and monocyte subpopulations were phenotyped by flow cytometry. Transcriptome profiling by RNA sequencing identified disease-associated gene signatures, which were validated by RT-qPCR. The CD14+ monocyte secretome was analysed using multiplex assays. Functional ability of CAVD-derived CD14+ monocytes to induce myofibroblastic transdifferentiation (MT) and osteoblastic differentiation (OD) of human valvular interstitial cells (VICS) was evaluated by immunocytochemistry and quantitative o-cresolphthalein complexone assays. ResultsIn PBMCs, CAVD monocytes displayed a subpopulation shift, with an increased proportion of CD14CD16- classical monocytes and a reduced CD14CD16 non-classical monocyte levels. In CD14+ monocytes, transcriptomic analysis revealed upregulation of inflammation-related (PDK4) and calcification-related (ATP2B1) genes, alongside downregulation of immunomodulatory genes (DDR1, IKBKE). Secretome analysis showed reduced production of immunomodulatory and anti-osteoblastogenic cytokines (IL-4, CCL3) while promoting gene expression of factors promoting MT and OD in VICS. These alterations were associated with a marked monocyte-induced increase in SMA and OPN expression in VICS and a two-fold increase in calcification. ConclusionWe demonstrate for the first time that circulating monocytes from patients with CAVD exhibit enhanced pro-inflammatory and pro-calcific properties that may contribute to CAVD progression. Additionally, we identify dysregulated gene sets within these monocytes that represent potential novel therapeutic targets for CAVD.

BackgroundFabry disease (FD) is a lysosomal storage disorder impacting multiple organs, including the heart. We investigated whether early-stage globotriaosylceramide (Gb3) accumulation, before occurrence of inclusion bodies, could cause significant stress and irreversible damages of the cardiomyocytes in FD patients. To assess the cellular stress and irreversible damage of cardiomyocytes in FD during early-stage Gb3 accumulation before the occurrence of typical pathology. MethodsImmunofluorescent (IF) staining or Western blotting were performed on fibroblasts from FD patients and myocardial biopsies from G3Stg/GLAko mice and FD patients. Notably, all biopsies exhibited detectable Gb3 accumulation under IF but lacked typical FD (Gb3 inclusion body) pathology. Staining targeted nuclear factor-{kappa}B (NF-{kappa}B), interleukin-18 (IL-18), phospho-p42/44 mitogen-activated protein kinase (MAPK), and inducible nitric oxide synthase (iNOS) as inflammatory and oxidative stress markers. Alpha-smooth muscle actin (-SMA) IF staining was conducted to detect myofibroblasts. ResultsFibroblasts from FD patients, in conjunction with cardiomyocytes from both G3Stg/GLAko mice and FD patients, exhibited significant accumulation of inflammatory markers such as NF-{kappa}B IL-18 and phospho-p42/44 MAPK, as well as the oxidative stress marker iNOS. Despite the absence of typical FD pathology, the presence of fibrosis was confirmed in myocardial biopsies from these patients through strong positive staining of -SMA. ConclusionsSignificant cellular stress and even irreversible damage may occur before the onset of typical pathological changes in cardiomyocytes of FD. Based on our findings, treatment should be initiated much earlier than we currently thought to prevent irreversible damage and improve the prognosis of FD patients.

IntroductionThis study investigates the immunohistochemical expression of hPG, ANXA2, and the polarization of tumor-associated macrophages (TAMs) in gastric adenocarcinomas. Methods: A retrospective analysis was conducted on FFPE tissue samples from 60 patients with gastric adenocarcinoma (primary tumors, lymph node metastases and normal-looking gastric mucosa), and on gastric biopsies from 23 healthy controls. The expression of hPG and ANXA2 was quantified using the H-score, and the CD163/HLA-DR ratio was used to infer macrophage polarization (M2/M1). Results: ANXA2 expression was significantly elevated in primary tumors and lymph node metastases compared to normal and healthy controls and increased with tumor grade. High ANXA2 expression was associated with poorer overall and disease-free survival. In contrast, hPG expression, although positively correlating to ANXA2 expression, showed no prognostic value. The M2/M1 ratio increased with tumor progression, showed a negative correlation with ANXA2 expression and failed to correlate significantly with survival. Conclusions: This study is the first to demonstrate the adverse prognostic impact of ANXA2 overexpression in gastric adenocarcinoma tissues from Caucasian patients, hinting to its potential utility as a prognostic biomarker and therapeutic target. Further large-scale studies could aid validate these findings and explore the therapeutic potential of targeting ANXA2 and modulating the TAM polarization.

IntroductionDegenerative disc disease (DDD) is a major cause of lower back pain (LBP). Key pathological processes of intervertebral disc (IVD) degeneration include extracellular matrix (ECM) degradation (including aggrecan loss), cartilage dehydration, and pathological ingrowth of blood vessels and nerve fibers. Neurotrophins and neuropeptides, such as nerve growth factor (NGF) and substance P (SP), play an essential role in LBP pathogenesis, neoinnervation, inflammation, and the maintenance of chronic pain. Materials and MethodsThirty-six young patients (mean age 36.00 [31.00, 42.50] years) with LBP associated with herniated discs and five healthy individuals were enrolled. IVD samples were collected during microdiscectomy. MRI-based Pfirrmann classification (2001) was used to assess the stages of disc degeneration. Histological grading was performed according to Sives criteria (2002). Histochemical staining (hematoxylin-eosin, Alizarin Red, Safranin O/Fast Green FCF) was conducted to evaluate ECM status, including aggrecan content. Immunohistochemical analysis was performed to assess NGF, S-100 protein, and SP expression. ResultsAll patients experienced chronic LBP. According to MRI, Pfirrmann grade V degeneration was found in 30.55% of patients, grade IV in 61.11%, grade III in 5.56%, and grade II in 2.78%. Histologically confirmed degeneration was observed in 23 cases (63.88%), with 3 patients showing severe degeneration (10-12 points). In patients with longer pain episodes (average duration 11.29 weeks), aggrecan loss was observed in 19.4% of cases (r=0.449; P=0.031). NGF expression was significantly higher in degenerated discs (P=0.0287) and positively correlated with SP levels (r=0.785; P=5.268 x 10-9). Increased NGF and SP expression were noted in patients with osteophytes, with levels correlating with both the histological degeneration score and MRI grading. Isolated free nerve endings were detected in the nucleus pulposus of 5 patients. Calcification was observed in 36.1% of cases, predominantly around hypertrophic chondrocytes and their clusters, and its severity correlated with radiculopathy (r=0.664; P=0.005). ConclusionIn young individuals, aggrecan loss, increased expression of NGF, SP, S-100 protein, and ECM calcification are key pathological features of IVD degeneration contributing to chronic LBP. The colocalization of NGF and SP suggests a synergistic role in the development of chronic pain. These findings highlight new therapeutic targets aimed at inhibiting pathological neoinnervation and ECM degradation.

This study aimed to examine the inhibitory effects of Euonymine on in-stent restenosis (ISR) after percutaneous coronary intervention (PCI) and oxidized low-density lipoprotein (ox-LDL)-induced proliferation, migration, and pro-apoptotic of vascular smooth muscle cells (VSMCs) in vitro, and its potential mechanisms. Euonymine is a monomer component extracted from Tripterygium hypoglaucum (Levl) Hutch. Using in vitro models of rabbit carotid balloon injury and porcine atherosclerotic coronary implantation, we confirmed that Euonymine inhibited ISR after PCI. Furthermore, Euonymine inhibited VSMC phenotypic transformation by targeting AKT1 to regulate the PTEN/AKT1/m TOR signaling pathway, with exertion of anti-proliferative, anti-migratory, and pro-apoptotic effects on ox-LDL-induced cell injury model. Additionally, the study demonstrated that Euonymine induced apoptosis of VSMCs via the p38MAPK-related mitochondria-dependent apoptotic pathway. Collectively, these findings indicated that Euonymine drug-eluting stents inhibited ISR after PCI by targeting AKT1 and p38MAPK to enhance the contractile phenotype of VSMCs to prevent intimal hyperplasia development. This provides insights into a potential therapeutic strategy involving the beneficial effect of Euonymine drug-eluting stent on ISR. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/466441v1_ufig1.gif" ALT="Figure 1"> View larger version (34K): org.highwire.dtl.DTLVardef@1582ef9org.highwire.dtl.DTLVardef@73e133org.highwire.dtl.DTLVardef@1c5f318org.highwire.dtl.DTLVardef@daca49_HPS_FORMAT_FIGEXP M_FIG C_FIG

IntroductionPlaque rupture is the primary trigger of the acute clinical manifestations of atherosclerotic disease. So far, factual insight in the processes leading up to cap destabilization is largely missing. In order to overcome this knowledge gap, a pseudo-timeline of atherosclerosis progression was established in order to systematically map the qualitative changes in cap characteristics during lesion progression and destabilization. Material and MethodsA pseudo-timeline was created by randomly selecting preclassified (revised AHA classification, at least 10 per stage) left coronary artery FFPE specimens obtained during tissue donation (aortic valve procurement). Qualitative changes were visualized by (immuno)histochemistry, immunofluorescence and confocal microscopy. Scoring was performed by two observers using semiquantitative scoring estimates. ResultsThe median age of the donors was 56 years (IQR 51.5-59), and 67% of the patients was male. Movat staining indicated a consistent pattern of cap formation, maturation and destabilization. A distinctive cap emerged in the early fibroatheroma stage of progressive atherosclerosis. Disease progression was accompanied by profound fibrotic changes in the gap, and a progressive presence of inanotic (nutritional deprivation leading to dissolution) mesenchymal cells. Plaque rupture was preceded by thinning of the collagen fibers and accumulation of foam cells in the central portion of the thin cap. No evidence was found for a direct involvement of neovascularization in the destabilization process. ConclusionThe pseudo-time line of atherosclerotic lesion development characterizes the development of an unstable cap as a degenerative and fibrotic process with progressive exhaustion of the mesenchymal cell population. This study provides a rationale for the limited efficacy of medical strategies aimed at plaque stabilization.

Glycosaminoglycans (GAGs) pooling has been considered since long as one of the histopathological characteristics defining thoracic aortic aneurysm (TAA) together with smooth muscle cells (SMCs) apoptosis and elastin fibers degradation. However, few information is provided about GAGs composition or potential implication in TAA pathology. Syndecan-1 (Sdc-1) is a heparan sulfate proteoglycan that is implicated in extracellular matrix (ECM) interaction and assembly, regulation of SMCs phenotype and various aspects of inflammation in the vascular wall. In the current work, the regulation of Sdc-1 protein was examined in human TAA by ELISA and immunohistochemistry. In addition, the role of Sdc-1 was evaluated in descending TAA in vivo using a mouse model combining both aortic wall weakening and hypertension. Our results showed that Sdc-1 protein is over expressed in human TAA aortas compared to healthy counterparts and that SMCs are the major cell type expressing Sdc-1. Similarly, in the mouse model used, Sdc-1 expression was increased in TAA aortas compared to healthy samples. Although its protective role against abdominal aneurysm has been reported, we observed that Sdc-1 was dispensable for TAA prevalence or rupture. In addition, Sdc-1 deficiency did not alter the extent of aortic wall dilatation, elastin degradation, collagen deposition, or leukocyte recruitment in our TAA model. These findings suggest that Sdc-1 could be a biomarker revealing TAA pathology. Future investigations could uncover the underlying mechanisms leading to Sdc-1 expression alteration in TAA.

Pregnancy in young-onset Parkinsons disease (YOPD) is rare; however, medication for this condition is critical for maternal and fetal health. Tolcapone is an effective antiparkinsonian drug. However, its safety studies on mothers and fetuses are limited. In this study, we aimed to investigate the effects of tolcapone on the mother and the developing fetus during different stages of gestation. Tolcapone was administered to pregnant mice at a dose of 60 (H-Tol) or 30 mg/kg/day (L-Tol) during early or mid-gestation. We observed that tolcapone administration during early gestation causes abortion and delays fetal development in a dose-dependent manner. During mid-gestation tolcapone barely caused embryo lethality; however, the mice developed preeclampsia-like phenotypes, including maternal hypertension, proteinuria, and fetal growth restriction. Histomorphological analysis of placentas from tolcapone treated mice revealed abnormalities in the trophoblast layer and the impaired trophoblast invasion in the decidua. Mechanistically, we revealed that tolcapone inhibits the invasion and migration of trophoblasts in vitro, with changes in the protein expression of Snail, Twist, and E-cadherin. In conclusion, tolcapone caused embryo lethality and growth restriction during early gestation, whereas it caused preeclampsia-like phenotypes in mice with defective trophoblast invasion in mid-gestation. Collectively, our study provides novel insights into the effects of tolcapone on pregnancy.

BackgroundMelatonin (MEL) is an indole amine molecule primarily produced in the pineal gland. Melatonin has been shown in numerous studies to have anti-fibrosis characteristics in the kidney, liver, and other organs. However, it is still unclear how melatonin works in bladder fibrosis. We explored how melatonin affected animals with bladder fibrosis and its underlying mechanisms. Materials and MethodsMEL was used to treat human bladder smooth muscle cells (HBdSMCs) after they were stimulated with TGF-{beta}1 in vitro. Proteomic analysis and then bioinformatic analysis based on the alterations in these proteins were then performed on HBdSMCs from the different processing methods. To construct an in vivo bladder fibrosis model, we injected protamine sulfate (PS) and lipopolysaccharide (LPS) twice a week into the rat bladder for six weeks. After two weeks of PS/LPS treatment, the treatment group was treated with MEL (20mg/kg/d) for 4 weeks. Finally, we detected the expression of fibrosis markers from different perspectives. The TGF-{beta}1/Smad pathway, and EMT in cell and bladder tissues were also identified. Further proteomic analysis was also performed. ResultsIn the in vitro experiment, we found that TGF-{beta}1 treatment enhanced the fibrosis markers Collagen III and -SMA of HBdSMCs. E-cadherin expression decreased while TGF-{beta}1/Smad pathway was activated. Vimentin and N-cadherin expressions were also elevated at the same time. Similar findings were observed in the LPS group. After MEL treatment, the expression of collagen III and -SMA decreased, and the expression of E-cadherin increased, while the expression of Vimentin and N-cadherin also decreased. CCN1 and SQLE may be the important proteins in the development of bladder fibrosis, according to quantitative proteomics analysis. MEL can decrease their expressions which leadis to relief of bladder fibrosis. Bioinformatics analysis shows that the extracellular space structure related to metabolic pathways, actin filament binding, and stress fibers can serve as a pivotal focus in the management of fibrosis. ConclusionMelatonin attenuates bladder fibrosis by blocking the TGF-{beta}1/Smad pathway and EMT. CCN1 appears to be a possible therapeutic target for bladder fibrosis.

BackgroundSepsis-induced cardiomyopathy (SIC) poses a significant challenge in critical care, necessitating comprehensive understanding and innovative diagnostic approaches. This study explores the immune-related molecular intricacies underlying SIC, employing bioinformatics analyses and machine learning techniques. MethodsRNA-seq and scRNA-seq datasets (GSE79962 and GSE190856) were obtained from the Gene Expression Omnibus (GEO). After initial quality control and preprocessing, scRNA-seq data (GSE190856) were analyzed using the Seurat package, including cell clustering and annotation. The CellChat package was then used to analyze immune cell interactions. Unsupervised clustering of SIC patients was performed based on differentially expressed ICD-related genes (GSE79962). Immune cell infiltration and gene set variation analysis were conducted, and weighted gene co-expression network analysis identified co-expression modules. A predictive signature for SIC was constructed through machine learning methods. ResultsTrough analyzing the GSE190856 scRNA-seq dataset, the communication between macrophages/monocytes and lymphocytes was found to be enhanced in mouse myocardial tissue during the early onset of SIC. Meanwhile, the expression level of ICD-related genes was upregulated in the monocytes infiltrating to the heart. These results suggestted that ICD may play a crucial role in the pathogenesis of SIC, which had been verified by the upregulated expression of ICD-related genes in the hearts of SIC patients in the GSE79962 dataset. The SIC patients were classified to 2 clusters, with cluster 1 exhibited an upregulation of the renin-angiotensin system, while cluster 2 displayed heightened activity in the RIG-I-like receptor signaling pathway. After comparing four machine learning models, the support vector machine (SVM) model exhibited better discrimination for SIC patients. By correlating the expression levels of the five crucial genes contained in this model with the clinical features of SIC patients, we found that JARID2 was negatively related to the Left Ventricular Ejection Fractions, while TNIP2 was negatively related to the variety of inotropes and vasopressors used in the SIC patients. ConclusionThis research unveils the correlation between ICD and SIC, offering insights into immune activity in the hearts during sepsis. The constructed SVM model with selected genes provides a promising molecular strategy for SIC diagnosis.

BackgroundCervical cancer is one of the most common malignant tumors of the female reproductive system. Existing treatments provide limited benefit for patients with advanced, recurrent or metastatic disease, and reliable prognostic markers are lacking. In this study we integrated multi-omic data from The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) database. Protein-coding genes meeting the criteria of an adjusted P value < 0.05 and |log2 fold-change| > 5 were screened; 693 genes were identified. We further focused on three genes related to the tumor microenvironment--interleukin 21 (IL21), C-X-C motif chemokine ligand 9 (CXCL9) and cluster of differentiation 1A (CD1A)--and performed differential expression analysis, survival analysis, clinical stage analysis and immune infiltration correlation analysis to clarify their prognostic value and potential mechanisms in cervical cancer. Results(1) CXCL9 and CD1A were highly expressed in cervical cancer tissues, and all three genes showed high expression across different pathological stages without stage-dependent differences; (2) high expression of IL21, CXCL9 and CD1A improved patient prognosis and was positively associated with overall survival (OS), disease-specific survival (DSS) and progression-free interval (PFI); (3) expression of IL21, CXCL9 and CD1A was closely correlated with infiltration of multiple immune cells: IL21 correlated with total T cells, helper T cells and B cells, CXCL9 correlated with T cells and activated dendritic cells, and CD1A correlated with immature dendritic cells. ConclusionIL21, CXCL9 and CD1A are potential prognostic biomarkers and key immunomodulatory factors in cervical cancer. This study provides a new direction for immunotherapy and individualized precision treatment of cervical cancer.

Aims: This study aimed to explore the biological activities of miR-199a-5p in dextran sulphate sodium (DSS)-induced ulcerative colitis and apoptosis and identify the direct target of miR-199a-5p in this process. Main methods: HT-29 cells and C57BL/6 mice were used to examine the function of miR-199a-5p in vitro and in vivo, respectively. Expression of miRNA and mRNA was measured using quantitative real-time PCR and western blotting was used to measure the change in protein expression. Flow cytometry was subsequently employed to determine cell apoptosis, and a luciferase assay was used to confirm the direct target of miR-199a-5p. Results: Expression of miR-199a-5p was increased by DSS treatment in mice. In parallel, miR-199a-5p is found to be involved in endoplasmic reticulum stress (ERS) and cell apoptosis in HT-29 cells, and its upregulation induced ERS, apoptosis, weight loss, and ulcerative colitis in mice in vivo, which could be prevented by the suppression of miR-199a-5p. Luciferase assay confirmed that the 3' untranslated region (3'-UTR) of XBP1 is the target binding site of miR-199a-5p. Conclusion: miR-199a-5p promotes ulcerative colitis and cell apoptosis by targeting the 3'-UTR of XBP1. Our findings reveal a new regulatory mechanism for ERS signaling and suggest that miR-199a-5p might be a potential target for UC therapy.

PurposeThe bladder dysfunction associated with type 2 diabetes mellitus (T2DM) involves urine storage and voiding disorders. We evaluated the pathologic conditions of bladder wall in a rat model of T2DM and evaluated the effects of the phosphodiesterase-5 (PDE-5) inhibitor tadalafil (TA). Materials and MethodsMale Otsuka Long-Evans Tokushima Fatty (OLETF) rats and Long-Evans Tokushima Otsuka (LETO) rats comprised T2DM and control groups. TA was orally administered for 12 weeks. The bladder blood flow and ATP released from the bladder epithelium were measured using laser speckle imaging and an organ bath bladder distention test. The expression levels of markers of hypoxia, pro-inflammatory cytokines, and growth factors in the bladder wall were measured by real-time PCR and ELISA. The contractions of bladder strips in response to KCl and carbachol were monitored in OLETF rats. ResultsThe bladder blood flow was impaired and there was greater ATP release and vesicular nucleotide transporter (VNUT) expression in the OLETF rats than in the LETO rats, but these effects were suppressed by TA administration. Furthermore, the high expression of HIF-1, 8-OHdG, IL-6, TNF-, IGF-1, and bFGF in the OLETF rats was reduced by TA administration. In the OLETF rats, the contractile responses of bladder strips to KCl and carbachol were impaired, but were restored by TA administration. ConclusionsThe impairment of bladder blood flow in rats with T2DM is associated with greater ATP release and the upregulation of VNUT, markers of hypoxia, proinflammatory cytokines, and growth factors in the bladder epithelium. PDE5 inhibition has the potential to prevent the storage and voiding dysfunction associated with T2DM.