T cell exhaustion methylation signature drives differential immune responses in glioblastoma.

BACKGROUND: Methylation-related signatures play crucial roles in tumorigenesis and progression. However, their roles in the immune response in primary glioblastoma (GBM) remains unclear. METHODS: We analyzed the differential expression of specific members of T cell exhaustion-related pathways in GBM from the perspective of T cell exhaustion. We further screened for significantly negatively correlated methylation sites as candidate methylation markers for T cell exhaustion. Using consensus clustering, we divided the samples into two categories with significant differences in overall survival (OS). We then performed univariate and multivariate Cox regression analyses to construct the T Cell Exhaustion Methylation (TEXM) signature. Finally, we confirmed that this signature served as an independent prognostic factor, and further characterized it in terms of drug resistance and immunotherapy. RESULTS: We identified 95 significantly differentially expressed T cell exhaustion-related genes and 51 methylation markers associated with T cell exhaustion. The cancer samples were classified according to methylation site markers, thus indicating two subtypes with significant differences in OS: subtype A and subtype B. Tumor scores, stromal scores, tumor purity, and ESTIMATE scores all showed significant differences between subtypes (P < 0.05). Univariate Cox regression analysis identified five methylation sites significantly associated with OS, and multivariate Cox regression analysis was used to construct the TEXM signature model by using these five methylation sites. Significant differences in OS were found between the groups with high and low TEXM signature scores, on the basis of calculation of the TEXM signature scores of tumor samples and using the median score to divide them into high and low score groups. Survival analysis revealed that the high score group had poorer OS and DFS than the low score group in the validation set. Notably, we observed a significant difference in drug sensitivity between the high and low TEXM signature score groups, with the high score group showing higher drug resistance and poorer prognosis. The tumor immune state, as predicted with Tracking Tumor Immunophenotype (TIP), revealed significant differences in antitumor immune scores between the high and low TEXM signature score groups. Finally, we identified 43 significantly differentially regulated metabolism-associated biological processes. CONCLUSION: The epigenetic methylation-related TEXM signature plays a key role in driving differential immune responses in GBM.

SIRT6 alleviates senescence induced by Porphyromonas gingivalis in human gingival fibroblasts.

OBJECTIVE: Bidirectional influences between senescence and inflammation are newly discovered. This study aimed to clarify the roles and mechanism of Porphyromonas gingivalis (P. gingivalis) in exacerbating senescence in human gingival fibroblasts (HGFs). DESIGN: Subgingival plaque and gingivae were collected from twenty-four periodontitis patients and eighteen periodontally healthy subjects. Quantities of P. gingivalis in subgingival plaque were explored using real-time PCR and the expressions of p53, p21 and SIRT6 in gingivae were detected by IHC. Moreover, senescence in HGFs was induced by P. gingivalis lipopolysaccharide (LPS) and the expressions of senescence-related ß-galactosidase (SA-ß-gal), p53, p21 and senescence-associated secretory phenotype (IL-6 and IL-8) with or without treatment by SIRT6 activator UBCS039 were explored by IHC, western blot and ELISA, respectively. In addition, the levels of SIRT6, Nrf2, HO-1 and reactive oxygen species (ROS) were examined by western blot and flow cytometry. RESULTS: Quantities of P. gingivalis in subgingival plaque and semi-quantitative scores of p53 and p21 in gingivae of periodontitis patients were increased compared with healthy controls (p < 0.05), while SIRT6 score in periodontitis patients was decreased (p < 0.001). Quantities of P. gingivalis were positively correlated with p53 and p21 scores (0.6 < r < 0.9, p < 0.01), and negatively correlated with SIRT6 score (-0.9 < r<-0.6, p < 0.01). Moreover, P. gingivalis LPS increased the levels of SA-ß-gal, p53, p21, IL-6, IL-8 and ROS and decreased the levels of SIRT6, Nrf2 and HO-1 in HGFs, which was rescued by UBCS039 (p < 0.05). CONCLUSIONS: P. gingivalis LPS could induce senescence of HGFs, which could be reversed by SIRT6 via Nrf2-HO-1 signaling pathway.

Key Cell-in-Cell Related Genes are Identified by Bioinformatics and Experiments in Glioblastoma.

Purpose: This study aimed to explore the roles of cell-in-cell (CIC)-related genes in glioblastoma (GBM) using bioinformatics and experimental strategies. Patients and Methods: The ssGSEA algorithm was used to calculate the CIC score for each patient. Subsequently, differentially expressed genes (DEGs) between the CIClow and CIChigh groups and between the tumor and control samples were screened using the limma R package. Key CIC-related genes (CICRGs) were further filtered using univariate Cox and LASSO analyses, followed by the construction of a CIC-related risk score model. The performance of the risk score model in predicting GBM prognosis was evaluated using ROC curves and an external validation cohort. Moreover, their location and differentiation trajectory in GBM were analyzed at the single-cell level using the Seurat R package. Finally, the expression of key CICRGs in clinical samples was examined by qPCR. Results: In the current study, we found that CIC scorelow group had a significantly better survival in the TCGA-GBM cohort, supporting the important role of CICRGs in GBM. Using univariate Cox and LASSO analyses, PTX3, TIMP1, IGFBP2, SNCAIP, LOXL1, SLC47A2, and LGALS3 were identified as key CICRGs. Based on this data, a CIC-related prognostic risk score model was built using the TCGA-GBM cohort and validated in the CGGA-GBM cohort. Further mechanistic analyses showed that the CIC-related risk score is closely related to immune and inflammatory responses. Interestingly, at the single-cell level, key CICRGs were expressed in the neurons and myeloids of tumor tissues and exhibited unique temporal dynamics of expression changes. Finally, the expression of key CICRGs was validated by qPCR using clinical samples from GBM patients. Conclusion: We identified novel CIC-related genes and built a reliable prognostic prediction model for GBM, which will provide further basic clues for studying the exact molecular mechanisms of GBM pathogenesis from a CIC perspective.

Integrative analysis reveals the potential prognostic roles and immunological values of unc-5 netrin receptor A (UNC5A) in glioma.

BACKGROUND: UNC5A had been reported to play crucial roles in multiple cancers. However, little was known about the associations among UNC5A and glioma. Therefore, we first combined scRNA-seq, proteomics, as well as bulk RNA-seq in order to investigate UNC5A's functions in gliomas. METHODS: Online databases provided scRNA-seq, proteomics, as well as bulk RNA-seq data on UNC5A in gliomas. The following procedures were conducted in order: QRT-PCR, Norman chart, gene set enrichment analysis (GSEA), and univariate/multifactor Cox regression analyses. We further explored the associations among UNC5A and tumor immunity. RESULTS: By comparing gliomas with normal tissues, the TCGA dataset showed a significantly reduced expression of UNC5A, which was also confirmed by GSE50161, GSE4290, and QRT-PCR findings (p < 0.05). In both the TCGA and CGGA datasets, gliomas patients with low-UNC5A expression would have poorer overall survival (OS) prognoses (p < 0.05). ScRNA-seq analysis by the CancerSEA online website presented that UNC5A had a low expression in various glioma clusters and significantly associated with six functional states. Moreover, UNC5A might be a reliable independent biomarker of OS in gliomas patients (p < 0.05). Based on the results of GSEA, UNC5A might be connected to three significant pathways in gliomas. We also successfully created a Norman chart to assess the OS prognoses of these patients. Additionally, in aspects of tumor immunity, the infiltration levels of immune cells in LGG, the immune cell pathways, tumor immune microenvironment, as well as immune checkpoints in both LGG and GBM were revealed to be significantly influenced by UNC5A (p < 0.05). CONCLUSIONS: UNC5A was found to have prognostic and immunological significance in gliomas, offering patients with gliomas new treatment options.

Retraction: An MSN-PEG-IP drug delivery system and IL13Rα2 as targeted therapy for glioma.

Retraction of 'An MSN-PEG-IP drug delivery system and IL13Rα2 as targeted therapy for glioma' by Jinlong Shi et al., Nanoscale, 2017, 9, 8970-8981, https://doi.org/10.1039/C6NR08786H.

M2 macrophage marker CHI3L2 could serve as a potential prognostic and immunological biomarker in glioma by integrated single-cell and bulk RNA-Seq analysis.

BACKGROUND: CHI3L2 plays a crucial role in multiple cancers, but its importance in glioma remains unclear. Hence, we comprehensively integrated bulk RNA-sequencing (RNA-seq), proteomics and single-cell RNA-seq (scRNA-seq) to determine the roles of CHI3L2 in gliomas. METHODS: Bulk RNA-seq, proteomics and scRNA-seq data of CHI3L2 in glioma were obtained from online databases. The quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) were conducted to verify the CHI3L2 expression. Then, univariate and multivariate Cox regression analyses, Norman charts and gene set enrichment analysis (GSEA) were performed. Finally, the associations between CHI3L2 and tumor immunity were explored. RESULTS: The expression of CHI3L2 was markedly higher in glioma cancers compared with normal tissues from analysis of the data of the Cancer Genome Atlas and Chinese Glioma Genome Atlas datasets and as verified by GSE4290, GSE50161, qRT-PCR and IHC results (p < 0.05). High expression of CHI3L2 suggested poor overall survival (OS) prognosis in gliomas (p < 0.05). CHI3L2 might also serve as an independent predictor of OS for gliomas (p < 0.05) and we also constructed a Norman chart to predict these patients' survival prognosis with good performance. GSEA analysis showed that CHI3L2 might be involved with eight pathways in gliomas. Regarding tumor immunity, CHI3L2 was found to be significantly involved with immune cell infiltration levels of low-grade glioma, the tumor immune microenvironment, immune checkpoints and immune cells in both low-grade glioma and glioblastoma (p < 0.05). Additionally, scRNA-seq data for CHI3L2 in glioma from the TISCH2 website showed that CHI3L2 is mainly expressed in astrocytes, endothelial cells, CD8+ T cells, mono/macrophage cells, etc. CONCLUSIONS: CHI3L2 presents prognostic and immunological values in glioma, providing novel therapeutic targets for glioma patients.

A fault diagnosis method for building electrical systems based on the combination of variational modal decomposition and new mutual dimensionless.

The fault diagnosis of building electrical systems are of great significance to the safe and stable operation of modern intelligent buildings. In this paper, it has many problems, such as various fault types, inconspicuous fault characteristics, uncertainty of fault type and mode, irregularity, unstable signal, large gap between fault data classes, small gap between classes and nonlinearity, etc. A method of building electrical system fault diagnosis based on the combination of variational mode decomposition and mutual dimensionless indictor (VMD-MDI) and quantum genetic algorithm-support vector machine (QGA-SVM) is proposed. Firstly, the method decomposes the original signal through variational modal decomposition to obtain the optimal number of Intrinsic Mode Function(IMF) containing fault feature information. Secondly, extracts the mutual dimensionless indicator for each IMF. Thirdly, the optimal penalty coefficient C of the support vector machine and the parameter gamma ([Formula: see text]) in the radial basis kernel function are selected by the quantum genetic algorithm. Finally, SVM optimized by the QGA is used to identify and classify the faults. By applying the proposed method to the experimental platform data of building electrical system, and compared with the traditional feature extraction method Empirical Mode Decomposition (EMD), Singular Value Decomposition(SVD), Local Mean Decomposition(LMD). And compared with traditional SVM, Genetic Algorithm optimized Support Vector Machine (GA-SVM), One-Dimensional Convolutional Neural Network (1DCNN) for fault classification methods. The experimental results show that the method has better effect and higher accuracy in fault diagnosis and classification of building electrical system. Its average test accuracy can reach 91.67[Formula: see text].

Shape-programmable magneto-active elastomer composites for curve and biomimetic behavior imitation.

A programming methodology, which can be applied to soft-magnetic-material-based magneto-active elastomers (MAEs), to catch the predefined specific objective curves is proposed in this study. The objective curves have been equally separated into a couple of segments, which will be filled by the designed MAE elements. Furthermore, the designed MAE segments with different chain angles, in which the deformation orientation of each element under applied homogeneous magnetic fields has been investigated based on the designed experimental setup, are arrayed based on the proposed programming methodology to constitute the MAE composite to catch the orientation of the objective curve. The experimental results show that based on the proposed programming methodology, the MAE composites can describe different curves, which include harmonic, tangential and arc tangential functions under applied homogeneous magnetic fields with good agreement. Furthermore, on the basis of the proposed programming methodology, the MAE composites are utilized to mimic the typical biomimetic behavior (the peeking-up behavior of snakes and the flapping behavior of birds) with smooth curvature properties, in which the dynamic procedures present continuous curves.

Magnetoactive Soft Drivers with Radial-Chain Iron Microparticles.

Magnetoactive elastomers (MAEs), one kind of typical novel magnetoactive driver applied in the soft robotic area, have become one of the research hotspots as they can provide biologically friendly driving methods with safe, preprogrammed, and easy-to-implement properties. In this study, novel MAEs embedding soft magnetic iron microparticles with radial chains, which can be molded in one piece, achieve 3D deformation, and co-work between multiple MAEs under single homogeneous stimuli, are proposed. Then, two kinds of novel magnetoactive drivers are established based on the proposed MAEs, which can achieve the synchronous pumping behavior of heart and the extension behavior of muscle under applied homogeneous magnetic fields. The experimental data show that (1) for the pumping behavior, the maximum instantaneous flow rate and total pumping volume can reach 200.1 and 52.3 mL/min, respectively, under 120 BPM applied harmonic magnetic field with 0-300 mT amplitude; (2) the muscle extension behavior can achieve a strain of 0.925 without a loading mass and carry a load of 40 times its own weight with a pronounced dynamic movement. It should be emphasized that the behavior of the proposed magnetoactive drivers can be excited by remote homogeneous magnetic fields, and it has great application potential in biomimetic or bioinspired soft driving systems.

Combination Glioma Therapy Mediated by a Dual-Targeted Delivery System Constructed Using OMCN-PEG-Pep22/DOX.

Accumulating studies have investigated the efficacy of receptor-mediated delivery of hydrophobic drugs in glioma chemotherapy. Here, a delivery vehicle comprising polyethylene glycol (PEG) and oxidized nanocrystalline mesoporous carbon particles (OMCN) linked to the Pep22 polypeptide targeting the low-density lipoprotein receptor (LDLR) is designed to generate a novel drug-loaded system, designated as OMCN-PEG-Pep22/DOX (OPPD). This system effectively targets glioma cells and the blood-brain barrier and exerts therapeutic efficacy through both near-infrared (NIR) photothermal and chemotherapeutic effects of loaded doxycycline (DOX). Pathological tissue microarrays show an association of LDLR overexpression in human glioma tissue with patient survival.NIR irradiation treatment and magnetic resonance imaging results show that OPPD reaches the effective glioma-killing temperature in a glioma-bearing rat with a skull bone removal model and considerably reduces glioma sizes relative to the drug-loaded system without the Pep22 peptide modification and the control respectively. Thus, OPPD not only effectively targets LDLR-overexpressing glioma but also exerts a dual therapeutic effect by transporting DOX into the glioma and generating thermal effects with near-infrared irradiation to kill tumor cells. These collective findings support the utility of the novel OPPD drug-loaded system as a promising drug delivery vehicle for clinical application in glioma therapy.

An MSN-PEG-IP drug delivery system and IL13Rα2 as targeted therapy for glioma.

A combination of gene therapy and chemotherapy has recently received interest as a targeted therapy for glioma. A mesoporous silica nanoparticle (MSN)-based vehicle coated with IL13Rα2-targeted peptide (IP) using polyethylene glycol (PEG), MSN-PEG-IP (MPI), was constructed and confirmed as a potential glioma-targeted drug delivery system in vitro. In this work, tissue microarray (TMA) results revealed that IL13Rα2 was over-expressed in human glioma tissues and that high expression of IL13Rα2 in patients was associated with poor survival. Doxorubicin (DOX)-loaded MPI (MPI/D) crossed the blood-brain barrier, specifically targeting glioma cells and significantly enhancing the cellular uptake of DOX in glioma cells compared with MSN/DOX (M/D) and MSN-PEG/DOX (MP/D), whereas the normal brain was not affected. Magnetic Resonance Imaging (MRI) examinations showed that the tumour size of glioma-bearing rats in the MPI/D-treated group was much smaller than those in the M/D and MP/D treated groups. Immunofluorescence results demonstrated that MPI/D treatment induced more apoptosis and much less proliferation than the other two treatments. However, the therapeutic effect was weak when IL13Rα2 was knocked down. Furthermore, U87 cells treated with IL-13 and MPI together could increase both STAT6 and P63 expression, which attenuated glioma cell proliferation, invasion and migration compared with cells treated with IL-13 alone. The results of the subcutaneous tumour model also revealed that IL13Rα2 knockdown could hinder cell proliferation and induce more apoptosis. The promising results suggested that MPI can not only deliver DOX to glioma in a targeted manner but also occupy IL13Rα2, which can promote IL-13 binding to IL13Rα1 and activation of the JAK-STAT pathway to induce an anti-glioma effect.

Combined Effect of Sodium Selenite and Ginsenoside Rh2 on HCT116 Human Colorectal Carcinoma Cells.

BACKGROUND: Sodium selenite and ginsenoside Rh2 (G-Rh2) are well known for their anticancer properties and have been exploited as a new therapeutic approach. In this study, we are interested to evaluate if sodium selenite and G-Rh2 combination results in a synergistic anticancer effect that could contribute to lower systemic toxicity. METHODS: We observed the synergistic antitumor effect by combination of sodium selenite and G-Rh2 on HCT-116 human colorectal carcinoma cells in vitro. Cell growth, viability, cell cycle progression and cell apoptosis, Bax/Bcl2 ratio, caspase-3 expression, reactive oxygen species (ROS) production and autophagy were evaluated. RESULTS: The results showed that sodium selenite and G-Rh2 combination have a synergistic effect on cell growth inhibition (57%) compared with sodium selenite (25%) and G-Rh2 alone (28%) after 24 hours of treatment. This combination also induced G1 and S phase arrest simultaneously and increased apoptosis rate. The results also indicated that Bax/Bcl2 ratio and caspase-3 expression, known as proapoptotic factors, were increased in the presence of sodium selenite and G-Rh2 alone. However, combined drug treatment results in a more significant increase in Bax/Bcl2 ratio and caspase-3 expression (P < 0.05). In addition, this combination significantly induces a depletion of ROS production and autophagy, compared to control, sodium selenite and G-Rh2 alone (P < 0.05). CONCLUSION: Sodium selenite and ginsenoside Rh2 combination may be a more effective treatment for human colorectal carcinoma and is a promising chemotherapeutic approach for malignant tumors.

Esophageal replacement by hydroxylated bacterial cellulose patch in a rabbit model.

BACKGROUND/AIM: To repair esophageal defects by hydroxylated and kombucha-synthesized bacterial cellulose (HKBC) patch in a rabbit model. MATERIALS AND METHODS: Semicircular esophageal defects 1 cm in length of the cervical esophagus were initially created in 18 Japanese big-ear rabbits and then repaired with HKBC patch grafts. The clinical outcomes including survival rate, weight change, food intake, and hematological and radiologic evaluation were observed. After X-ray evaluation, the rabbits were sacrificed sequentially at 1, 3, and 6 months for histopathologic analysis with light microscopy and scanning electron microscopy. RESULTS: Survival rate during the first month was 88.9% (n = 16). Two rabbits died from anastomotic leakage during the entire follow-up. Postoperatively, feeding function and body weight were gradually restored in the surviving animals. No hematological abnormalities were found, and no obvious anastomotic leakage, stenosis, or obstruction was observed under X-ray examination. The histopathologic results showed a progressive regeneration of the esophagus in the graft area, where the neo-esophagus tissue had characteristics similar to native esophageal tissue after 3 months of surgery. CONCLUSION: HKBC is beneficial for esophageal tissue regeneration and may be a promising material for esophageal reconstruction.

2,3,7,8-Tetrachlorodibenzo-p-dioxin promotes astrocyte activation and the secretion of tumor necrosis factor-α via PKC/SSeCKS-dependent mechanisms.

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous environmental pollutant that could induce significant toxic effects in the human nervous system. However, the underlying molecular mechanism has not been entirely elucidated. Reactive astrogliosis has implicated in various neurological diseases via the production of a variety of pro-inflammatory mediators. Herein, we investigated the potential role of TCDD in facilitating astrocyte activation and the underlying molecular mechanisms. We showed that TCDD induced rapid astrocyte activation following TCDD exposure, which was accompanied by significantly elevated expression of Src-Suppressed-C Kinase Substrate (SSeCKS), a protein involved in protein kinase C (PKC)-mediated Nuclear Factor kappa B signaling, suggesting a possible involvement of PKC-induced SSeCKS activation in TCDD-triggered reactive astroglia. In keeping with the finding, we found that the level of phosphorylated Nuclear Factor kappa B p65 was remarkably increased after TCDD treatment. Furthermore, interference of SSeCKS attenuated TCDD-induced inducible nitric oxide synthase, glial fibrillary acidic protein, phospho-p65 expression, and tumor necrosis factor-α secretion in astrocytes. In addition, pre-treatment with PKC inhibitor also attenuated TCDD-induced astrocyte activation, as well as SSeCKS expression. Interestingly, we found that TCDD treatment could lead to SSeCKS perinuclear localization, which could be abolished after treatment with PKC inhibitor. Finally, we showed that inhibition of PKC activity or SSeCKS expression would impair TCDD-triggered tumor necrosis factor-α secretion. Our results suggested that TCDD exposure could lead to astrocyte activation through PKC/SSeCKS-dependent mechanisms, highlighting that astrocytes might be important target of TCDD-induced neurotoxicity. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) elicits neurotoxic effects. Here, we show TCDD induces pro-inflammatory responses in astrocytes. TCDD initiates an increase of [Ca2+]i, followed by the activation of PKC, which then induces the activation of Src-suppressed C-kinase substrate (SSeCKS). SSeCKS promotes NF-κB activation and the secretion of TNF-α and nitric oxide in astrocytes.