Identification of Key Genes in Atherosclerosis by Combined DNA Methylation and miRNA Expression Analyses.

BACKGROUND: Atherosclerosis is a significant cause of coronary heart disease, cerebral infarction, and peripheral vascular disease. The objective of this study was to identify the key genes aberrantly expressed in atherosclerosis, which were regulated by microRNAs and DNA methylation. METHODS: We acquired data on DNA methylation and microRNA and messenger RNA expression from Gene Expression Omnibus data sets (GSE46394, GSE53675, and GSE12288, respectively) and identified differentially methylated genes, differentially expressed genes, and differentially expressed microRNAs between atherosclerosis and control samples. The miRDB, miRTarBase, and TargetScan databases were used to predict differentially expressed microRNAs-targeted genes, which were then intersected with differentially methylated genes and differentially expressed genes to identify genes associated with aberrant DNA methylation and microRNA activity. The DAVID database was used to perform functional enrichment analysis of differentially methylated genes and the key genes involved in atherosclerosis. Potential therapeutic agents for atherosclerosis were predicted by Connectivity Map analysis. RESULTS: In total, we identified 47 upregulated hypomethylated and 90 downregulated hypermethylated genes in atherosclerosis. Among them, 24 differentially expressed genes were found to be modulated both through aberrant DNA methylation and microRNA expression, and 10 such differentially expressed genes were defined as the key genes in atherosclerosis. Fifteen chemicals were selected for their potential effect in . CONCLUSIONS: We identified 10 key genes significantly associated with aberrant DNA methylation and microRNA expression in atherosclerosis and suggested 15 chemicals with potential effects on these genes, which could be further investigated as candidate drugs for atherosclerosis.

Association Between Telomere Length and Skin Cancer and Aging: A Mendelian Randomization Analysis.

Background: Telomere shortening is a hallmark of cellular senescence. However, telomere length (TL)-related cellular senescence has varying effects in different cancers, resulting in a paradoxical relationship between senescence and cancer. Therefore, we used observational epidemiological studies to investigate the association between TL and skin cancer and aging, and to explore whether such a paradoxical relationship exists in skin tissue. Methods: This study employed two-sample Mendelian randomization (MR) to analyze the causal relationship between TL and skin cancer [melanoma and non-melanoma skin cancers (NMSCs)] and aging. We studied single nucleotide polymorphisms (SNPs) obtained from pooled data belonging to genome-wide association studies (GWAS) in the literature and biobanks. Quality control was performed using pleiotropy, heterogeneity, and sensitivity analyses. Results: We used five algorithms to analyze the causal relationship between TL and skin aging, melanoma, and NMSCs, and obtained consistent results. TL shortening reduced NMSC and melanoma susceptibility risk with specific odds ratios (ORs) of 1.0344 [95% confidence interval (CI): 1.0168-1.0524, p = 0.01] and 1.0127 (95% CI: 1.0046-1.0209, p = 6.36E-07), respectively. Conversely, TL shortening was validated to increase the odds of skin aging (OR = 0.96, 95% CI: 0.9332-0.9956, p = 0.03). Moreover, the MR-Egger, maximum likelihood, and inverse variance weighted (IVW) methods found significant heterogeneity among instrumental variable (IV) estimates (identified as MR-Egger skin aging Q = 76.72, p = 1.36E-04; melanoma Q = 97.10, p = 1.62E-07; NMSCsQ = 82.02, p = 1.90E-05). The leave-one-out analysis also showed that the SNP sensitivity was robust to each result. Conclusion: This study found that TL shortening may promote skin aging development and reduce the risk of cutaneous melanoma and NMSCs. The results provide a reference for future research on the causal relationship between skin aging and cancer in clinical practice.

Neutrophil extracellular traps induce a hypercoagulable state in glioma.

BACKGROUND: Venous thromboembolism (VTE) is one of the leading complications in glioma patients. Neutrophil extracellular traps (NETs) have been reported to play a critical role in the physiopathology of cancer. We aimed to investigate the presence and potential role of NETs in the hypercoagulable state in glioma patients. Moreover, we evaluated the interaction between NETs and endothelial cells (ECs) in glioma patients. METHODS: The plasma levels of NETs were detected by enzyme-linked immunosorbent assay. The NET procoagulant activity was performed based on fibrin formation assays. The NET generation and NET-treated ECs in vitro were observed by confocal microscopy. Activated platelets (PLTs) and PLT-neutrophil aggregates were detected by flow cytometry. RESULTS: Plasma NET markers were significantly higher in stage III/IV glioma patients than in stage I/II glioma patients and healthy subjects. PLTs from glioma patients tended to induce NET formation than those from healthy subjects. NETs contributed to the hypercoagulable state in glioma patients. After ECs were incubated with NETs isolated from stage III/IV glioma patients, they lost their intercellular connections and were converted into procoagulant phenotypes. Combining DNase I and activated protein C markedly decreased endothelial dysfunction. CONCLUSIONS: Our results showed the interaction between NETs and hypercoagulability in glioma patients. Targeting NETs may be a potential therapeutic and prevention direction for thrombotic complications in glioma patients.

Neutrophil extracellular traps induce thrombogenicity in severe carotid stenosis.

BACKGROUND: Severe carotid stenosis is a common cause of stroke. In addition, previous clinical studies revealed that patients symptomatic of carotid stenosis suffer from increased episodes of stroke compared with their asymptomatic counterparts. However, the mechanism underlying these differences in the recurrence of stroke remains unclear. OBJECTIVE: The present study aimed to evaluate the levels of neutrophil extracellular traps (NETs) in the plasma of patients with severe carotid stenosis and investigate whether NETs induced procoagulant activity (PCA) in severe carotid stenosis. The study also sought to investigate the interactions between platelets or endothelial cells (ECs) and NETs. METHODS: The levels of NETs in plasma were quantified using enzyme-linked immunosorbent assay (ELISA). In addition, NETting neutrophils and neutrophil-platelet aggregates were detected through flow cytometry. On the other hand, the morphology of NETs formation and endothelial cells were analyzed through confocal microscopy. Finally, the procoagulant activity (PCA) of NETs and endothelial cells were assessed through ELISA and fibrin formation. RESULTS: Patients with symptomatic carotid stenosis patients had significantly higher levels of NETs markers compared with their asymptomatic counterparts and healthy subjects. In addition, increased levels of neutrophil-platelet aggregates induced the generation of NETs in patients with symptomatic carotid stenosis. Moreover, NETs contributed to PCA through tissue factor (TF), in patients with carotid stenosis. Furthermore, NETs disrupted the endothelial barrier and converted endothelial cells (ECs) into PCA to enhance the PCA in patients with carotid stenosis. CONCLUSIONS: The current study revealed differences in the levels of NETs in the plasma of symptomatic and asymptomatic patients suffering from carotid stenosis. The study also uncovered the interaction between NETs and thrombogenicity in carotid stenosis. Therefore, inhibiting NETs may be a potential biomarker and therapeutic target for recurring stroke in severe carotid stenosis.

Hierarchical assembly of dual-responsive biomineralized polydopamine-calcium phosphate nanocomposites for enhancing chemo-photothermal therapy by autophagy inhibition.

The induction of autophagy in cancer cells would occur in response to several therapy strategies, including chemotherapy and photothermal therapy (PTT). Hence, combined autophagy inhibition has been regarded as a prevailing strategy to enhance treatment sensitivity in cancers. Herein, dual pH/thermal responsive biomineralized nanocomposites (PCNPs) were rationally designed and prepared based on the hierarchical assembly of calcium phosphate (CaP) and polydopamine (PDA). The first step in the self-assembly process involves the incorporation of hydrophobic chemotherapeutic docetaxel (DTX) into the CaP nanoparticles. Next, PDA was utilized as the coating to hierarchically self-assemble onto the surface of CaP through a simple self-polymerization of dopamine. Third, the autophagy inhibitor chloroquine (CQ) was absorbed onto the surface of PDA via non-covalent interactions, forming PCNPs/DC. CQ was the only FDA approved autophagy inhibitor in clinical trials that could inhibit autophagosome fusion and degradation. The resulting PCNPs/DC could exhibit dual pH/thermal responsive properties due to the acid-sensitive CaP core and the photothermal effect of the PDA coating. Effective inhibition of autophagy in cancer cells could be realized by blocking the lysosome and weakening the degradation of autolysosomes by PCNPs/DC. Interestingly, complementary autophagy inhibition could therefore sensitize the effects of chemo-photothermal therapy both in vitro and in vivo with negligible toxicity. Therefore, these hierarchically assembled biomineralized nanocomposites would be used as a prevailing strategy to sensitize chemo-photothermal therapy by complementary autophagy inhibition.

A non-conjugated polyethylenimine copolymer-based unorthodox nanoprobe for bioimaging and related mechanism exploration.

Unconventional non-conjugated photoluminescent polymers have attracted increasing attention in bioimaging application, however their nonclassical photoluminescence mechanisms remain largely unclear. Herein, an amphiphilic copolymer polyethyleneimine-poly(d,l-lactide) (PEI-PDLLA) was synthesized and the obtained PEI-PDLLA copolymer exhibited intrinsic visible blue luminescence in the solid and concentrated solution states under 365 nm UV light irradiation. Using a computational assay approach, we investigated the unconventional photoluminescence mechanism of PEI-PDLLA. The results revealed that such photoluminescence should be related to the "clustered heteroatom chromophores" formed by through-space electronic interactions of N-heteroatoms in PEI. The copolymers can function as a fluorescent nanoprobe (PEI-PDLLA NPs) via a facile nanoprecipitation method and the self-assembly mechanism of PEI-PDLLA NPs was also investigated in-depth by molecular dynamics simulation. Intriguingly, the PEI-PDLLA NPs exhibited a remarkable excitation-dependent multi-wavelength emission characteristic, which was promising in acquiring a high precision imaging effect. Moreover, in contrast with conventional organic dyes with aggregation-caused quenching (ACQ), the fluorescence intensity of the PEI-PDLLA NPs was enhanced with increasing solution concentration. Furthermore, their applications in bioimaging indicated that PEI-PDLLA NPs could be utilized as a lysosome-specific and tumor-targeted nanoprobe with excellent photostability and good biocompatibility.

Isoacteoside, a dihydroxyphenylethyl glycoside, exhibits anti-inflammatory effects through blocking toll-like receptor 4 dimerization.

BACKGROUND AND PURPOSE: Isoacteoside (is a phenylethanoid isolated from Monochasma savatieri Franch. ex Maxim., which is an anti-inflammatory herb widely used in traditional Chinese medicine. However, the exact mechanism of the anti-inflammatory activity of isoacteoside is not completely understood. In this study, its anti-inflammatory mechanism was elucidated in mouse macrophages. EXPERIMENTAL APPROACH: The expression of the NF-κB pathway, MAPK pathway, iNOS, TNF-α, IL-6 and IL-1ß was evaluated using Western blotting, quantitative real-time PCR or ELISA. TLR4 dimerization was determined by transfecting HEK293T cells with TLR4 plasmids. The in vivo anti-inflammatory effect of isoacteoside was determined using mouse models of xylene-induced ear oedema, LPS-induced endotoxic shock and LPS-induced endotoxaemia-associated acute kidney injury (AKI). KEY RESULTS: Isoacteoside suppressed COX-2, iNOS, TNF-α, IL-6 and IL-1ß expression. Furthermore, isoacteoside attenuated the LPS-induced transcriptional activity of NF-κB by decreasing the levels of phosphorylated IκB-α and IKK and NF-κB/p65 nuclear translocation. In addition, isoacteoside inhibited LPS-induced transcriptional activity of AP-1 by reducing the levels of phosphorylated JNK1/2 and p38MAPK. Isoacteoside blocked LPS-induced TLR4 dimerization, resulting in a reduction in the recruitment of MyD88 and TIR-domain-containing adapter-inducing interferon-ß (TRIF) and the phosphorylation of TGF-ß-activated kinase-1 (TAK1). Pretreatment of mice with isoacteoside effectively inhibited xylene-induced ear oedema and LPS-induced endotoxic death and protected against LPS-induced AKI. CONCLUSIONS AND IMPLICATIONS: Isoacteoside blocked TLR4 dimerization, which activates the MyD88-TAK1-NF-κB/MAPK signalling cascades and TRIF pathway. Our data indicate that isoacteoside is a potential lead compound for the treatment of inflammatory diseases.