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Abstract Objective To investigate the association between circulating secretoneurin (SN) and angiographic coronary collateralization in stable angina patients with chronic coronary total occlusion (CTO). Methods SN concentrations in serum were measured in 641 stable angina patients with CTO by radioimmunoassay. The status of coronary collaterals from the contra-lateral vessel was visually estimated using the Rentrop grading system, and was categorized into poor (grade 0 or 1) or good (grade 2 or 3) collateralization. Results Serum SN levels were significantly higher in patients with good coronary collaterals compared to those with poor collaterals (175.23 ± 52.09 pmol/L vs. 143.29 ± 42.01 pmol/L, P

As the main body of resource aggregation, virtual power plant (VPP) not only needs to participate in the external energy market, but also needs to optimize the management of internal resources.Different from other energy storage systems, hydrogen energy storage systems can participate in the hydrogen market in addition to the backup and supplement functions of electric energy.For a virtual power plant operator (VPPO), it is necessary to optimize the dispatch of internal resources as well as to develop a bidding strategy for both the electricity and hydrogen markets based on operational conditions and external market information.In this study, a two-stage model was developed, taking into account the interaction mechanism between the internal and external sides.The first stage of the model was based on the operational optimization of the internal resources of renewable energy, flexible load, pumped storage and hydrogen storage systems, while the second stage of the model aimed to optimize the bidding strategy by maximizing the total revenue in the electricity energy market, auxiliary services market and hydrogen market.

ABSTRACTCytolethal distending toxins (CDTs) are released by Gram-negative pathogens into the extracellular medium as free toxin or associated with extracellular vesicles (EVs), commonly known as outer membrane vesicles (OMVs). CDT production by the gastrointestinal pathogen Campylobacter jejuni has been implicated in colorectal tumorigenesis. Despite CDT being a major virulence factor for C. jejuni, little is known about the EV-associated form of this toxin. To address this point, C. jejuni mutants lacking each of the three CDT subunits (A, B, and C) were generated. C. jejuni cdtA, cdtB, and cdtC bacteria released EVs in similar numbers and sizes to wild-type bacteria, ranging from 5 to 530 nm (mean ± SEM = 118 ±6.9 nm). As the CdtAC subunits mediate toxin binding to host cells, we performed “surface shearing” experiments, in which EVs were treated with proteinase K and incubated with host cells. These experiments indicated that CDT subunits are internal to EVs and that surface proteins are probably not involved in EV-host cell interactions. Furthermore, glycan array studies demonstrated that EVs bind complex host cell glycans and share receptor binding specificities with C. jejuni bacteria for fucosyl GM1 ganglioside, P1 blood group antigen, sialyl, and sulfated Lewisx. Finally, we show that EVs from C. jejuni WT but not mutant bacteria induce cell cycle arrest in epithelial cells. In conclusion, we propose that EVs are an important mechanism for CDT release by C. jejuni and are likely to play a significant role in toxin delivery to host cells.IMPORTANCECampylobacter jejuni is the leading cause of foodborne gastroenteritis in humans worldwide and a significant cause of childhood mortality due to diarrheal disease in developing countries. A major factor by which C. jejuni causes disease is a toxin, called cytolethal distending toxin (CDT). The biology of this toxin, however, is poorly understood. In this study, we report that C. jejuni CDT is protected within membrane blebs, known as extracellular vesicles (EVs), released by the bacterium. We showed that proteins on the surfaces of EVs are not required for EV uptake by host cells. Furthermore, we identified several sugar receptors that may be required for EV binding to host cells. By studying the EV-associated form of C. jejuni CDT, we will gain a greater understanding of how C. jejuni intoxicates host cells and how EV-associated CDT may be used in various therapeutic applications, including as anti-tumor therapies.

Abstract Contaminants within cell culture media often co‐isolate with eukaryotic extracellular vesicles (EVs) thus affecting their biological properties. It has yet to be investigated if this is also true for bacterial EVs (BEVs), especially for organisms grown in complex culture media containing animal‐derived products. To address this question, we isolated BEVs from the fastidious bacterium Helicobacter pylori grown in either standard Brain Heart Infusion (BHI) medium or BHI depleted of animal‐derived products (D‐BHI). We show that BEVs prepared from bacteria grown in D‐BHI medium have similar morphologies, size ranges and yields to those prepared from standard medium. Similarly, no differences were found in the ability of H. pylori BEVs to induce IL‐8 responses in epithelial cells. However, H. pylori BEVs prepared from D‐BHI medium were of higher purity than those prepared from standard medium. Importantly, proteomic analyses detected 3.4‐fold more H. pylori proteins and 10‐fold fewer bovine‐derived proteins in BEV samples prepared from D‐BHI rather than the standard method. Fifty‐seven H. pylori proteins were uniquely detected in BEV samples prepared from D‐BHI. In conclusion, we have described an improved method for BEV isolation. Furthermore, we demonstrate how animal‐derived products in bacteriological culture media may adversely affect proteomic analyses of BEVs.

Abstract The activity of integrin-linked kinase (ILK) in cancerous cells is often oncogenic and associated with malignant properties, such as uncontrolled cell cycle progression and evasion from senescence. However, the role of ILK in cellular senescence in gastric cancer (GC) has not been previously examined. We generated single-cell clones of ILK knock-out using CRISPR-Cas9 in human GC lines with mesenchymal or epithelial histology. Cells with no residual ILK expression exhibited strong cellular senescence with diminished clathrin-mediated endocytosis, Surprisingly, ILK loss-induced cellular senescence appeared to be independent of its function in integrin signaling. The low dose of CPD22, a small molecule inhibitor of ILK activity-induced senescence in three GC cell lines with different histologies. Furthermore, senescent cells with ILK depletion transfected with N-terminal truncated ILK mutant remaining catalytic domains displayed the reduction of senescent phenotypes. RNA sequencing and cytokine array results revealed the enrichment of multiple pro-inflammatory signaling pathways in GC lines in the absence of ILK. Our study identified the important role and the potential mechanism of ILK in the cellular senescence of cancerous epithelial cells. The inhibition of ILK activity using small molecule compounds could have a pro-senescent effect as a therapeutic option for GC.

Gastric cancer (GC), a leading cause of cancer-related death, is a heterogeneous disease. We aim to describe clinically relevant molecular classifications of GC that incorporate heterogeneity and provide useful clinical information. We combined different gene expression datasets and filtered a 7-gene signature related to the extracellular matrix (ECM), which also exhibited significant prognostic value in GC patients. Interestingly, putative CCCTC-binding factor (CTCF) regulatory elements were identified within the promoters of these ECM-related genes and were confirmed by chromatin immunoprecipitation sequencing (ChIP-Seq). CTCF binding sites also overlapped with histone activation markers, indicating direct regulation. In addition, CTCF was also correlated with the Wnt signaling pathway. A comparison of human GC cell lines with high or low expression of ECM-related genes revealed different levels of tumor aggressiveness, suggesting the cancer development-promoting functions of ECM-related genes. Furthermore, CTCF regulated COL1A1 and COLA31 expression in vitro. Silencing CTCF or COL1A1/COL1A3 markedly inhibited cell growth and migration in the metastatic GC cell line BGC823. Collectively, this ECM-related 7-gene signature provides a novel insight for survival prediction among GC patients. The zinc finger protein CTCF regulates ECM-related genes, thereby promoting GC cell growth and migration.

PurposeThe aim is to investigate the impacts of using multiplex immunochemistry (mIHC) staining to analyses the co-expression of programmed death ligand-1 (PD-L1) and tumor infiltrating lymphocytes (TILs) [CD8+ T cells and Forkhead Box Protein 3 (FOXP3)+ regulatory T cells (Tregs)] in different oral diseases, and oral squamous cell carcinoma (OSCC).MethodsFormalin fixed paraffin-embedded tissue sections from different oral diseases were stained with PD-L1 and TILs (CD8+ T cells and FOXP3+ Tregs) by mIHC staining simultaneously. The whole slide was scanned digitally to observe the cell phenotypes stained in the microenvironment. The contents of each slice were read using a computer-aided method to analyze and the cell densities were calculated using statistical software.ResultsWe were able to characterize the tumor microenvironment (TME) of different oral diseases including oral leukoplakia (OLK), inflammatory gingiva (IG), oral lichen planus (OLP), and squamous cell carcinoma (SCC), with accurate visualization of various immune cells harboring complex immune phenotypes by mIHC staining. The results showed that PD-L1 was up-regulated in SCC tissues at different pathological stages, while CD8 and FOXP3 had no significant changes. The ratio of PD-L1/CD8 was also significantly up-regulated in SCC tissues compared with that of other oral diseases. In advanced stages of OSCC, the FOXP3/CD8 ratio increased, and immunosuppressive TME was more pronounced. In addition, we also found different immune phenotypes: the inflamed phenotype, immune-excluded phenotypes, and immune-desert phenotypes. By locating tumor epithelial cells, we found that PD-L1 expression is in both tumor cells and stromal cells.ConclusionsmIHC is useful for the visualization and evaluation of tumor microenvironment in immuno-oncology research. It allows single-cell imaging in situ and could effectively and quickly determine the immune phenotype of different oral diseases.

Abstract Background Understanding immune phenotypes and human gastric disease in situ requires an approach that leverages multiplexed immunohistochemistry (mIHC) with multispectral imaging to facilitate precise image analyses. Methods We developed a novel 4-color mIHC assay based on tyramide signal amplification that allowed us to reliably interrogate immunologic checkpoints, including programmed death-ligand 1 (PD-L1), cytotoxic T cells (CD8+T) and regulatory T cells (Foxp3), in formalin-fixed, paraffin-embedded tissues of various human gastric diseases. By observing cell phenotypes within the disease tissue microenvironment, we were able to determine specific co-localized staining combinations and various measures of cell density. Results We found that PD-L1 was expressed in gastric ulcer and in tumor cells (TCs), as well as in tumor-infiltrating immune cells (TIICs), but not in normal gastric mucosa or other gastric intraepithelial neoplastic tissues. Furthermore, we found no significant reduction in CD8+T cells, whereas the ratio of CD8+T:Foxp3 cells and CD8+T:PD-L1 cells was suppressed in tumor tissues and elevated in adjacent normal tissues. An unsupervised hierarchical analysis also identified correlations between CD8+T and Foxp3+ tumor-infiltrating lymphocyte (TIL) densities and average PD-L1 levels. Three main groups were identified based on the results of CD8+T:PD-L1 ratios in gastric tumor tissues. Furthermore, integrating CD8+T:Foxp3 ratios, which increased the complexity for immune phenotype status, revealed 6–7 clusters that enabled the separation of gastric cancer patients at the same clinical stage into different risk-group subsets. Conclusions Characterizing immune phenotypes in human gastric disease tissues via multiplexed immunohistochemistry may help guide PD-L1 clinical therapy. Observing unique disease tissue microenvironments can improve our understanding of immune phenotypes and cell interactions within these microenvironments, providing the ability to predict safe responses to immunotherapies.

Abstract Objectives To facilitate disease prognosis and improve precise immunotherapy of gastric cancer (GC) patients, a comprehensive study integrating immune cellular and molecular analyses on tumor tissues and peripheral blood was performed. Methods The association of GC patients’ outcomes and the immune context of their tumors was explored using multiplex immunohistochemistry (mIHC) and transcriptome profiling. Potential immune dysfunction mechanism/s in the tumors on the systemic level was further examined using mass cytometry (CyTOF) in complementary peripheral blood from selected patients. GC cohorts with mIHC and gene expression profiling data were also used as validation cohorts. Results Increased CD4+FOXP3+ T‐cell density in the GC tumor correlated with prolonged survival. Interestingly, CD4+FOXP3+ T cells had a close interaction with CD8+ T cells rather than tumor cells. High densities of CD4+FOXP3+ T cells and CD8+ T cells (High‐High) independently predicted prolonged patient survival. Furthermore, the interferon‐gamma (IFN‐γ) gene signature and PDL1 expression were up‐regulated in this group. Importantly, a subgroup of genomically stable (GS) tumors and tumors with chromosomal instability (CIN) within this High‐High group also had excellent survival. The High‐High GS/CIN tumors were coupled with increased frequencies of Tbet+CD4+ T cells and central memory CD4+ T cells in the peripheral blood. Conclusion These novel findings identify the combination of CD8+ T cells and FOXP3+CD4+ T cells as a significant prognostic marker for GC patients, which also could potentially be targeted and applied in the combination therapy with immune checkpoint blockades in precision medicine.

Teacher feedback is the key to online collaborative discussion. To investigate the effects of different forms of teacher feedback intervention on learners' cognitive and emotional interactions in online collaborative discussion, this study collected collaborative discussion text data of online collaborative learners. Based on the framework of Community of Inquiry theory, naive Bayes algorithm for automatic coding of collaborative discussion text data was adopted. A bivariate (with or without emotion/guidance) experiment was designed based on teacher feedback. The participants of this study were college students (N = 109, average age = 20) of normal major participating in Teaching System Design. They were randomly divided into four experimental groups. In each experimental group, 4-5 people work in a group for collaborative learning. This study adopts quasi experimental research method, and the experiment period is 10 class hours. Reliability analysis, automatic text coding and ANOVA of cognitive-affective variables were used to conduct process mining for the collaborative discussion of four groups of learners by using heuristic mining algorithms. It found that different forms of teacher feedback have different effects on learners' cognitive emotion. Teachers' emotional feedback promotes learners' emotional interaction and cognitive interaction, whiccoch is easier to promote learners' cognitive interaction. Different forms of teacher feedback promote four types of cognitive emotion interaction process. This suggests that the multi-branch type of voice prompt feedback group has the best effect on learners' cognitive and emotional impact.

Current studies aiming at identifying single immune markers with prognostic value have limitations in the context of complex antitumor immunity and cancer immune evasion. Here, we show how the integration of several immune markers influences the predictions of prognosis of gastric cancer (GC) patients. We analyzed Tissue Microarray (TMA) by multiplex immunohistochemistry and measured the expression of immune checkpoint molecule PD-L1 together with antitumor CD8 T cells and immune suppressive FOXP3 Treg cells in a cohort of GC patients. Unsupervised hierarchical clustering analysis of these markers was used to define correlations between CD8 T, FOXP3 Treg and PD-L1 cell densities. We found that FOXP3 and PD-L1 densities were elevated while CD8 T cells were decreased in tumor tissues compared to their adjacent normal tissues. However, patient stratification based on each one of these markers individually did not show significant prognostic value on patient survival. Conversely, combination of the ratios of CD8/FOXP3 and CD8/PD-L1 enabled the identification of patient subgroups with different survival outcomes. As such, high densities of PD-L1 in patients with high CD8/FOXP3 and low CD8/PD-L1 ratios correlated with increased survival. Collectively, this work demonstrates the need for the integration of several immune markers to obtain more meaningful survival prognosis and patient stratification. In addition, our work provides insights into the complex tumor immune evasion and immune regulation by the tumor-infiltrating effector and suppressor cells, informing on the best use of immunotherapy options for treating patients.

In the present study, polysaccharides named ALPS and ASPS were isolated from Ampelopsis grossedentata leaves and stems, respectively. Physicochemical properties, in vitro antioxidant activities and the inhibitory effects on α-glucosidase of ALPS and ASPS were investigated. It was found that both ALPS and ASPS were acid protein-bound heteropolysaccharides, although with considerably different chemical composition and molecular weight distribution. Meanwhile, in comparison with ALPS, ASPS exhibited stronger antioxidant activity and inhibitory potential against α-glucosidase according to the in vitro evaluation. Moreover, our results suggested that protein and uronic acid might, at least partly, contribute positively to the biological behavior of ALPS and ASPS.

With the progress of science and technology, DCS has been widely used in the field of power plant automation. It integrates modern computer technology, modern control technology, modern communication technology and CRT technology to realize the monitoring and control of power production process. DCS network adopts 10Mbps Ethernet, which is the data transmission channel of DCS system. Its stability is related to the safe operation of the whole power plant. Therefore, more and more power generation enterprises begin to attach importance to network security management of DCS. In this paper, the author puts forward his own views on the network failure phenomenon of Foxboro I/A Series system, and gives the solution, hoping to have some reference significance for the safety management work of other similar DCS network of the unit.

At present, the flexible transformation of thermal power is an effective measure to solve the problem of renewable energy consumption. At the same time, the improvement of the electricity auxiliary service market makes the power generation enterprises improve their ability to provide auxiliary services through flexible transformation and obtain benefits while serving the power system. Therefore, how to construct the flexibility evaluation system of regional units according to the characteristics of the auxiliary service market in different regions is of great significance to power generation enterprises. In this paper, the flexibility evaluation system of the auxiliary market environment is designed. Analytic hierarchy process and fuzzy comprehensive evaluation method were used to determine the weight and final evaluation basis.

This study examines the clinical efficacy of high-dose naloxone in comatose emergency patients, focusing on its ability to enhance brain metabolism and reduce oxidative stress. A total of 120 patients were randomly assigned to a control group, which received conventional naloxone doses, and a study group, which received higher doses. Key outcomes measured included clinical efficacy, time to awakening, blood gas indices, inflammatory factors, consciousness level, neurological recovery and adverse effects. The study group showed a higher response rate (96.67% vs. 83.33%), regained consciousness more quickly, and had better blood gas indices and glasgow coma scale (GCS) scores (p<0.05). Neurological function recovery was superior in the study group, with fewer adverse reactions (6.67% vs. 20.00%, p<0.05). These results suggest that high-dose naloxone significantly improves treatment outcomes, enhancing wakefulness, reducing inflammation, and improving prognosis in emergency comatose patients, making it a promising option for clinical use. [ABSTRACT FROM AUTHOR]