EAF2 Downregulation Recruits Tumor-associated Macrophages in Prostate Cancer through Upregulation of MIF.

BACKGROUND: The role of tumor inflammatory microenvironment in the advancement of cancer, particularly prostate cancer, is widely acknowledged. ELL-associated factor 2 (EAF2), a tumor suppressor that has been identified in the prostate, is often downregulated in prostate cancer. Earlier investigations have shown that mice with EAF2 gene knockout exhibited a substantial infiltration of inflammatory cells into the prostatic stroma. METHODS: A cohort comprising 38 patients who had been diagnosed with prostate cancer and subsequently undergone radical prostatectomy (RP) was selected. These patients were pathologically graded according to the Gleason scoring system and divided into two groups. The purpose of this selection was to investigate the potential correlation between EAF2 and CD163 using immunohistochemistry (IHC) staining. Additionally, in vitro experimentation was conducted to verify the relationship between EAF2 expression, macrophage migration and polarization. RESULTS: Our study demonstrated that in specimens of human prostate cancer, the expression of EAF2 was notably downregulated, and this decrease was inversely associated with the number of CD163-positive macrophages that infiltrated the cancerous tissue. Cell co-culture experiments revealed that the chemotactic effect of tumor cells towards macrophages was intensified and that macrophages differentiated into tumor-associated macrophages (TAMs) when EAF2 was knocked out. Additionally, the application of cytokine protein microarray showed that the expression of chemokine macrophage migration inhibitory factor (MIF) increased after EAF2 knockout. CONCLUSIONS: Our findings suggested that EAF2 was involved in the infiltration of CD163-positive macrophages in prostate cancer via MIF.

Experimental study on visual ergonomics of an aircraft cockpit considering an extremely wide range of illuminance conditions.

Vision is the main way for pilots to obtain information, and good visual ergonomics are an important support for ensuring aircraft flight safety. The range of illumination changes in the light environment of the aircraft cockpit is very wide, and research on the visual ergonomics of the cockpit needs to consider various extreme lighting conditions. This study conducted visual ergonomics experiments on 15 participants in a full-scale simulated cockpit, examining the accuracy, reaction time, and subjective evaluation of visual tasks under 8 typical environmental lighting intensity levels. The experimental results show that, except for head-up display, the accuracy of visual target interpretation tasks performed by other display devices under different brightness conditions remains at a high level. And as the brightness of the display device increases, the accuracy of interpretation gradually increases, and the reaction time gradually decreases. In terms of subjective evaluation, there is a significant correlation between fuzziness, fatigue, clarity of image symbols, resolution between symbols, comfort of the image, and overall satisfaction with the image, but the correlation with environmental illumination level is relatively low. The experimental results can provide a certain theoretical basis for the design of cockpit lighting environment.

Prognostic value and drug sensitivity of F­box and leucine­rich repeat protein 6 in glioma.

Gliomas are highly malignant and invasive tumors lacking clear boundaries. Previous bioinformatics and experimental analyses have indicated that F-box and leucine-rich repeat protein 6 (FBXL6), a protein crucial for the cell cycle and tumorigenesis, is highly expressed in certain types of tumors. The high expression level of FBXL6 is reported to promote tumor growth and adversely affect patient survival. However, the molecular mechanism, prognostic value and drug sensitivity of FBXL6 in glioma remain unclear. To address this, the present study analyzed FBXL6 expression in gliomas, utilizing data from The Cancer Genome Atlas and Chinese Glioma Genome Atlas databases. Analysis of FBXL6 mRNA expression levels, combined with patient factors such as age, sex and tumor grade using Kaplan-Meier plots and nomograms, demonstrated a strong correlation between FBXL6 expression and glioma progression. Co-expression networks provided further insights into the biological function of FBXL6. Additionally, using CIBERSORT and TISDB tools, the correlation between FBXL6 expression correlation tumor-infiltrating immune cells and immune genes was demonstrated to be statistically significant. These findings were validated by examining FBXL6 mRNA and protein levels in glioma tissues using various techniques, including western blot, reverse transcription-quantitative PCR and immunohistochemistry. These assays demonstrated the role of FBXL6 in glioma progression. Furthermore, drug sensitivity analysis demonstrated a strong correlation between FBXL6 expression and various drugs, which indicated that FBXL6 may potentially act as a future promising therapeutic target in glioma treatment. Therefore, the present study identified FBXL6 as a diagnostic and prognostic marker in patients with gliomas and highlighted its potential role in glioma progression.

Tumor Exosomal ENPP1 Hydrolyzes cGAMP to Inhibit cGAS-STING Signaling.

To evade immune surveillance, tumor cells express ectonucleotide pyrophosphatase phosphodiesterase 1 (ENPP1) on the surface of their membrane, which degrades extracellular cyclic GMP-AMP (cGAMP), thereby inhibiting the cyclic GMP-AMP synthase (cGAS) stimulator of interferon gene (STING) DNA-sensing pathway. To fully understand this tumor stealth mechanism, it is essential to determine whether other forms of ENPP1 with hydrolytic cGAMP activity also are present in the tumor microenvironment to regulate this innate immune pathway. Herein, it is reported that various tumor-derived exosomes carry ENPP1, and can hydrolyze synthetic 2'3'-cGAMP and endogenous 2'3'-cGAMP produced by cells to inhibit cGAS-STING pathway in immune cells. Moreover, tumor exosomal ENPP1 also can hydrolyze 2'3'-cGAMP bound to LL-37 (an effective transporter of 2'3'-cGAMP) to inhibit STING signaling. Furthermore, high expression of ENPP1 in exosomes is observed isolated from human breast and lung cancer tissue, and tumor exosomal ENPP1 inhibited the immune infiltration of CD8+ T cells and CD4+ T cells. The results elucidate the essential function of tumor exosomal ENPP1 in the cGAS-STING pathway, furthering understanding of the crosstalk between the tumor cells and immune system.

Novel applications of histopathological markers to distinguish prognostic subgroups in colorectal adenocarcinoma.

OBJECTIVE: To explore the novel applications of histological factors by stratifying the prognostic markers of the overall CRC patients in subgroups. MATERIALS AND METHODS: A total of 17 histopathological and molecular factors were retrospectively collected and systematically analyzed for the prediction of CRC prognosis in the overall and stratified subgroups by using the Kaplan-Meier curve analysis as well as the Cox regression test. The χ2 test was used to analyze the correlation of the prognostic markers with other factors. RESULTS: The histopathological markers including the lymph node metastasis (LNM), perineural/venous invasion (PVI), TNM stage, the local recurrence or distant metastasis after surgery (R/M) and the molecular markers Ki-67 expression as well as KRAS mutation were identified to be the independent prognostic biomarkers in the overall CRC. The differential prognosis of LNM was found to be significant in age, tumor site, histological classification (histo_classification), cell differentiation, and KRAS/NRAS/BRAF (KNB) mutation stratified subgroups. The PVI was discovered to differently predict survival for patients in age, histo_classification, differentiation, and R/M stratified subgroups. Same as LNM and PVI, TNM was also found to demonstrate differential prognosis in age, tumor site, histo_classification, differentiation, R/M status and KRAS/KNB mutation stratified subgroups. More importantly, R/M was firstly identified not to be terrible for patients in age, histo_classification, LNM, TNM, Ki-67, and KRAS/KNB stratified subgroups. Besides, KRAS mutation was innovatively found to show differential prognosis in age, differentiation, and LNM stratified subgroups. CONCLUSIONS: The stratification analyses of prognostic markers in CRC patients indicate novel applications of the above histopathological and molecular markers in clinic and the findings provide new insights into future investigations of precision pathology.

The pathological markers LNM, PVI, TNM stage, R/M, the histological marker Ki-67 expression and the molecular marker KRAS mutation are all the early biomarkers capable of independently predicting the 2-year survival rate for CRC.Differential prognosis of the histopathological and molecular markers is commonly found in age, tumor site, differentiation, histological type, LNM, TNM, and R/M stratified CRC subgroups.

Significance of cuproptosis- related genes in the diagnosis and classification of psoriasis.

Cuproptosis is a novel form of cell death linked to mitochondrial metabolism and is mediated by protein lipoylation. The mechanism of cuproptosis in many diseases, such as psoriasis, remains unclear. In this study, signature diagnostic markers of cuproptosis were screened by differential analysis between psoriatic and non-psoriatic patients. The differentially expressed cuproptosis-related genes (CRGs) for patients with psoriasis were screened using the GSE178197 dataset from the gene expression omnibus database. The biological roles of CRGs were identified by GO and KEGG enrichment analyses, and the candidates of cuproptosis-related regulators were selected from a nomogram model. The consensus clustering approach was used to classify psoriasis into clusters and the principal component analysis algorithms were constructed to calculate the cuproptosis score. Finally, latent diagnostic markers and drug sensitivity were analyzed using the pRRophetic R package. The differential analysis revealed that CRGs (MTF1, ATP7B, and SLC31A1) are significantly expressed in psoriatic patients. GO and KEGG enrichment analyses showed that the biological functions of CRGs were mainly related to acetyl-CoA metabolic processes, the mitochondrial matrix, and acyltransferase activity. Compared to the machine learning method used, the random forest model has higher accuracy in the occurrence of cuproptosis. However, the decision curve of the candidate cuproptosis regulators analysis showed that patients can benefit from the nomogram model. The consensus clustering analysis showed that psoriasis can be grouped into three patterns of cuproptosis (clusterA, clusterB, and clusterC) based on selected important regulators of cuproptosis. In advance, we analyzed the immune characteristics of patients and found that clusterA was associated with T cells, clusterB with neutrophil cells, and clusterC predominantly with B cells. Drug sensitivity analysis showed that three cuproptosis regulators (ATP7B, SLC31A1, and MTF1) were associated with the drug sensitivity. This study provides insight into the specific biological functions and related mechanisms of CRGs in the development of psoriasis and indicates that cuproptosis plays a non-negligible role. These results may help guide future treatment strategies for psoriasis.

The pro-invasive factor COL6A2 serves as a novel prognostic marker of glioma.

Background: Glioma is an incurable malignant lesion with poor outcome characterized by easy recurrence after surgery with or without radiotherapy and chemotherapy. Studies have shown that COL6A2 is closely related to the tumorigenesis and development of a variety of tumors. However, the role of COL6A2 in glioma and the relationship between COL6A2 and tumor infiltrating immune cells remain unclear. Methods: Western blot, real-time PCR, a tissue microarray and immunohistochemistry were applied to detect COL6A2 mRNA and protein amounts in glioma, and all experiments were repeated three times. A tissue microarray of glioma samples was used for prognostic analysis. Detection of COL6A2 co-expression with immune genes using immunohistochemical methods, and tumor modeling using nude mice for prevention and treatment studies. Based on the mRNA expression of COL6A2, patients with glioma in TCGA were divided into the low and high COL6A2 expression groups, and GO and KEGG pathway analyses were performed. A PPI network was constructed using STRING, and the associations of COL6A2 with tumor-infiltrating immune cells and immune genes were analyzed in the CIBERSORT and TISIDB databases. COL6A2 mRNA and protein amounts were increased in glioma. Results: Multiple-database and tissue microarray analyses showed that COL6A2 expression in glioma was associated with poor prognosis, Tissue microarray showed that COL6A2 was the highest expressed in WHO IV and significantly higher in TCGA-GBM than in TCGA-LGG. Immunohistochemistry can well demonstrate the co-expression of COL6A2 with immune genes in a tumor model established in nude mice, showing that interference with COL6A2 expression may have an inhibitory effect on tumors. The mRNA expression of COL6A2 was involved in 22 KEGG pathways, and GSEA analysis showed that 28 and 57 gene sets were significantly enriched at nominal p values <0.01 and <0.05, respectively, protein network revealed a tight interaction between COL6A2 and SPARC. The CIBERSORT database indicated that COL6A2 was correlated with 15 types of tumor-infiltrating immune cells, including M2 macrophages, CD8 T cells, neutrophils, gamma delta T cells, activated CD4 memory T cells, follicular helper T cells, M0 macrophages, M1 macrophages, regulatory T cells (Tregs), activated NK cells, eosinophils, activated mast cells, monocytes, activated dendritic cells, and resting CD4 memory T cells. The TISIDB database indicated that COL6A2 was significantly correlated with lymphocytes such as regulatory T cell, Type 17 T helper cell, Type 1 T helper cell, and immunomodulatory genes. In addition, COL6A2-related immune regulatory genes show that most immune regulatorygenes have prognostic value for glioma, and high-risk immune genes are notconducive to the survival of glioma patients. Conclusions: COL6A2-related immune regulatory genes show that most immune regulatory genes have prognostic value for glioma, and high-risk immune genes are not conducive to the survival of glioma patients. COL6A2 may be a novel potential prognostic biomarker of glioma and associated with tumor-infiltrating immune cells in the tumor microenvironment, and interference with COL6A2 expression can inhibit tumor growth, which suggests COL6A2 as a potential target for future treatment.

A novel melanoma prognostic model based on the ferroptosis-related long non-coding RNA.

Ferroptosis is an iron-dependent programmed cell death related to the biological process of many kinds of tumors. Long noncoding RNAs (LncRNA) have been found to play essential roles in the tumor, and their functions in the ferroptosis of tumor cells have been partially discovered. However, there is no summary of ferroptosis-related LncRNA and its functions in melanoma. In the present study, we aim to explore the expression profile of ferroptosis-related LncRNA genes and their value in melanoma prognosis by bioinformatics analysis. The expression of ferroptosis-related gene (FRG) from melanoma clinical data was extracted based on the Cancer Genome Atlas (TCGA) database. By screening the RNA expression data of 472 cases of melanoma and 810 cases of normal skin, eighteen ferroptosis-related differential genes were found to be related to the overall survival rate. Furthermore, 384 ferroptosis-related LncRNAs were discovered through constructing the mRNA-LncRNA co-expression network, and ten of them were found with prognostic significance in melanoma by multivariate Cox analysis. Risk assessment showed that the high expression of LncRNA00520 is associated with poor prognosis, while the increased expression of the other LncRNA is beneficial to the prognosis of patients with melanoma. From univariate and multivariate Cox regression analysis, there were ten ferroptosis-related LncRNA risk models towards to be significant independent prognostic factors for patients with melanoma and valuable predictive factors for overall survival (OS)(P<0.05). The ROC curve further suggested that the risk score has relatively reliable predictive ability (AUC=0.718). The protein level of ferroptosis-related genes was verified by the HPA database and IHC test, leading to the discovery that the expressions of ALOX5, PEBP1, ACSL4, and ZEB1 proteins up-regulated in tumor tissues, and existed differences between tumor tissues and normal tissues. In conclusion, we identified ten ferroptosis-related LncRNA and constructed a prognosis model base.

Cell origin and genome profile difference of penoscrotum invasive extramammary Paget disease compared with its in situ counterpart.

Penoscrotum extramammary Paget disease (pEMPD) is a rare cutaneous carcinoma with an unknown cell origin. pEMPD always presents as a tumor in situ with an indolent process, whereas some progress into invasive forms with more aggressive behavior. The in situ and invasive cases display different morphologies and biological behavior, and thus far, a relationship between these two components has not been demonstrated. Immunohistochemistry was used to disclose the immunotype of pEMPD, and the results revealed that invasive/in situ pEMPD possessed with some identical immunophenotypes such as CK7, P63, and CK10, which inferred the clonal relatedness. The variable expressions of GCDFP-15 and carcino embryonic antigen hinted that tumor cell origin might be an epidermal sweat gland in epiderma. In our cohort, invasive pEMPD presented increased expression of androgen receptor and decreased MUC5CA expression, and these two changes might bring to the shift of invasive phenotype. To better understanding the relationship between these distinct tumor forms, we performed whole exome sequencing testing to evaluate overlapping genomic alterations of six paired invasive/in situ pEMPDs. The results showed that missense mutation was the predominant mutation type, and C>T transition accounted for 65.1% in all SNP mutation. Among the top 20 differential genes obtained from the six paired invasive/in situ pEMPD analysis, MUC4 (one missense, one in frame del, and one multi-hit), AHNAK2 (two missense and one multi-hit), DOT1L (two missense and one multi-hit), and FRG1 (two missense and one-multi hit) mutations were most enriched in invasive pEMPDs, which postulated that these genes may play roles in the disease progression.

Molecular Dynamics Simulation and Gas Generation Tracking of Pyrolysis of Bituminous Coal.

To study the generation rules of organic molecules or fragments and the generation paths of some hydrocarbon gases (C2H2/C2H4) and inorganic gases (CO2/H2O/H2/H2S) in the pyrolysis process of bituminous coal at 1000-5000 K, the ReaxFF molecular dynamics module in AMS software was used to simulate the pyrolysis behavior of the Hongqingliang model, Gaojialiang model, and Wiser model. With the increase of temperature, the system potential energy decreases, the endothermic efficiency increases first and then decreases, the fragments of C1-C4 fragments increase, and the gas molecules generated increase. In the pyrolysis process, the oxygen-containing functional groups and hydrogen groups formed H2O and H2 with the increase of temperature. H2S as an intermediate product is always maintained in dynamic equilibrium. CO2 comes from the decarboxylation of the carboxyl groups. When the temperature is lower than 3000 K, C2H4 and C2H2 are mainly formed by the adjacent carbon structure in coal molecules, and C2H4 is formed from the ethyl side chain, the naphthenic structure, and the unstable aromatic rings. C2H2 is formed from naphthene structures and aromatic rings with multiple side chains. When the temperature is higher than 3000 K, C2H4 and C2H2 are mainly formed by the random combination of free radicals generated by the crushing of coal molecules. The research results are of great significance to coal pyrolysis and clean utilization of coal.

Multiomics Landscape Uncovers the Molecular Mechanism of the Malignant Evolution of Lung Adenocarcinoma Cells to Chronic Low Dose Cadmium Exposure.

Cadmium (Cd) from cigarette smoke and polluted air can lead to lung adenocarcinoma after long-term inhalation. However, most studies are based on short-term exposure to this toxic metal at high concentrations. Here, we investigate the effects of long-term exposure of A549 cells (lung adenocarcinoma) to cadmium at low concentrations using morphological and multiomics analyses. First, we treated A549 cells continuously with CdCl2 at 1µM for 8 months and found that CdCl2 promoted cellular migration and invasion. After that, we applied transmission electron and fluorescence microscopies and did not observe significant morphological changes in Golgi apparatus, endoplasmic reticulum, lysosomes, or mitochondria on Cd treated cells; microfilaments, in contrast, accumulated in lamellipodium and adhesion plaques, which suggested that Cd enhanced cellular activity. Second, by using whole-exome sequencing (WES) we detected 4222 unique SNPs in Cd-treated cells, which included 382 unique non-synonymous mutation sites. The corresponding mutated genes, after GO and KEGG enrichments, were involved mainly in cell adhesion, movement, and metabolic pathways. Third, by RNA-seq analysis, we showed that 1250 genes (784 up and 466 down), 1623 mRNAs (1023 up and 591 down), and 679 lncRNAs (375 up and 304 down) were expressed differently. Furthermore, GO enrichment of these RNA-seq results suggested that most differentially expressed genes were related to cell adhesion and organization of the extracellular matrix in biological process terms; KEGG enrichment revealed that the differentially expressed genes took part in 26 pathways, among which the metabolic pathway was the most significant. These findings could be important for unveiling mechanisms of Cd-related cancers and for developing cancer therapies in the future.

Experimental study on the explosion characteristics of hydrogen-methane premixed gas in complex pipe networks.

To explore the overpressure evolution laws and flame propagation characteristics in complex pipe networks after the addition of hydrogen to methane, we experimentally studied the explosive pressure wave and flame wave propagation laws for three different premixed gas mixtures with hydrogen-methane concentrations of 0, 10% and 20% when the equivalence ratio was 1. Experimental results indicate that the maximum explosion overpressure of the premixed gas increases with increasing distance from the explosion source, and it shows a gradually decreasing trend. In the complex pipe network, an overpressure zone is formed in the B-E-H and D-E sections of the network. The flame temperature is superimposed with the superimposition of the pressure, showing a trend of first increasing, then decreasing, then increasing, and finally decreasing in the complex pipe network. The flame arrival time increases with increasing distance, and the maximum flame speed shows a decreasing trend. The peak overpressure and maximum flame velocity of the premixed gas under a hydrogen volume fraction of 20% are 1.266 MPa and 168 m/s. The experimental research results could provide important theoretical guidelines for the prevention and control of fuel gas explosions in urban pipe networks.

DNase I-assisted 2'-O-methyl molecular beacon for amplified detection of tumor exosomal microRNA-21.

An end-modified 2'-O-methyl molecular beacon (eMB) with unique nuclease resistance was designed and prepared. The eMB can resist the enzymatic digestion by DNase I, which would otherwise occur upon the hybridization of the eMB with a complementary sequence. As a result, the coupling use of eMBs and DNase I allows highly sensitive detection of miRNA with a limit of detection (LOD) of 2.5 pM. The analytical strategy was further used for detection of tumor exosomal microRNA-21, and down to 0.86 µg mL-1 A375 exosomes were detected. Overall, the present method can effectively quantify tumor-derived exosomes for cancer diagnosis.

Influence of acetylene on methane-air explosion characteristics in a confined chamber.

To study the impact of acetylene on methane explosions, the safe operation of coal mines should be ensured. In this paper, a 20 L spherical tank was used to study the explosive characteristics of acetylene-methane-air mixture. In addition, the GRI-Mech3.0 mechanism was used to study the chemical kinetic mechanism for the mixed gas, and the effect of adding acetylene on the sensitivity of methane and the yield of free radicals was analysed. The results show that acetylene can expand the scope for methane explosion, lower the lower explosion limit, and increase the risk of explosion. Acetylene increases the maximum explosion pressure, laminar combustion rate and maximum pressure rise rate for the methane-air mixture while shortening the combustion time. Three combustion modes for the acetylene-methane-air mixture were determined: methane-dominated, transitional and acetylene-dominated combustion modes. Chemical kinetic analysis for the mixed gas shows that as the volume fraction of acetylene increases, the generation rate for key free radicals (H*, O* and OH*) gradually increases, thereby increasing the intensity of the explosive reaction. The results from this research will help formulate measures to prevent coal mine explosion accidents.

Molecular simulation of gases competitive adsorption in lignite and analysis of original CO desorption.

To study the adsorption characteristics of CO, CO2, N2, O2, and their binary-components in lignite coal, reveal the influence of CO2 or N2 injection and air leakage on the desorption of CO in goafs, a lignite model (C206H206N2O44) was established, and the supercell structure was optimized under temperatures of 288.15-318.15 K for molecular simulation. Based on molecular dynamics, the Grand Canonical Monte Carlo method was used to simulate the adsorption characteristics and the Langmuir equation was used to fit the adsorption isotherms of gases. The results show that for single-components, the order of adsorption capacity is CO2 > CO > O2 > N2. For binary-components, the competitive adsorption capacities of CO2 and CO are approximate. In the low-pressure zone, the competitive adsorption capacity of CO2 is stronger than that of CO, and the CO is stronger than N2 or O2. From the simulation, it can be seen that CO2, N2 or O2 will occupy adsorption sites, causing CO desorption. Therefore, to prevent the desorption of the original CO in the goaf, it is not suitable to use CO2 or N2 injection for fire prevention, and the air leakage at the working faces need to be controlled.