Ruifang Sun

Millions of people chew areca nut to boost work efficiency and relieve stress. Areca nut contains arecoline, which adversely affects reproductive function, although the underlying molecular mechanisms remain unclear. Herein, we evaluated arecoline-mediated reproductive toxicity using Sprague–Dawley (SD) rat models and an in vitro GC-1 cell model. Molecular docking, quantitative reverse transcription PCR (qRT-PCR), Western blot, cell-based assays, and functional rescue experiments were used to investigate the mechanisms involved. Dose-dependent spermatogenic damage was observed in male rats receiving 50, 100 and 200 mg/kg arecoline, with reduced sperm counts and quality, higher testicular MDA and lower activity of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). Molecular docking predicted a possible interaction between arecoline and Notch1. qRT-PCR and Western blot analyses showed that the expression of Notch1 and its ligands, Jagged-1 and Dll-1, was downregulated in testicular tissues and GC-1 cells following arecoline exposure. Notch1 expression was strongly positively correlated with seminiferous tubule diameter and seminiferous epithelium height. In vitro, arecoline induced ROS accumulation, inhibited proliferation and promoted apoptosis in GC-1 cells. Rescue experiments showed that the Notch activator valproic acid, used to enhance Notch signaling, alleviated arecoline-induced toxicity, whereas Notch inhibitor FLI-06 worsened cell injury. These findings support a role for altered Notch signaling and oxidative stress in arecoline-induced spermatogenic injury and provide insight into the molecular mechanisms of its reproductive toxicity.

Background: Irritable bowel syndrome (IBS) is a prominent functional gastrointestinal disorder, yet the precise causes and mechanisms behind it remain largely unclear. Numerous environmental compounds have been associated with the intestinal health of individuals suffering from IBS. This study sought to explore the impact of environmental chemicals on the condition of IBS. Methods: We analyzed genome-wide association study (GWAS) data comprising 455,321 individuals of white British descent, among which 28,518 individuals with IBS underwent transcriptome-wide association study (TWAS) analysis. Reference gene expression data were sourced from tissues including the small intestine, transverse colon, sigmoid colon, whole blood, and peripheral blood. Results: Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted utilizing the significant genes identified through TWAS. Additionally, protein-protein interaction network analysis was performed using the STRING database to elucidate the functions of the proteins encoded by these genes. Furthermore, chemical-related gene set enrichment analysis (CGSEA) was employed to explore the associations between environmental chemicals and IBS. In total, TWAS identified 33 significant genes ( P _FDR < 0.05), while CGSEA revealed 112 chemicals significantly correlated with IBS ( P _FDR < 0.05, |NES| > 1). Both TWAS (targeting genetic influences) and CGSEA (focusing on environmental influences) were instrumental in pinpointing chemicals associated with IBS. Conclusion: The results of this study enhance our comprehension of the genetic and environmental determinants associated with functional gastrointestinal disorders.

This objective was to conduct Mendelian randomization (MR) analysis in order to investigate the potential causal relationship between chloride intracellular channel protein 5 (CLIC5) and Hashimoto thyroiditis (HT). Two separate MR analyses were performed, both using CLIC5 as the exposure variable and HT as the outcome. The first analysis utilized exposure data from genome-wide association study (GWAS) ID: prot-a-584 and outcome data from GWAS ID: ebi-a-GCST90018635. The second analysis used the same exposure data but sourced the outcome from ebi-a-GCST90018855. Four MR methods were applied in both analyses: MR Egger, weighted median, inverse variance weighted, and weighted mode. In both analyses, the inverse variance weighted method produced the most significant results, indicating a positive association between CLIC5 and HT. The first analysis estimated a causal effect of 0.4169 ( P = .005344), while the second analysis estimated a slightly lower effect of 0.1196 ( P = .0009933). The weighted median and weighted mode also suggested a positive association, although with lower statistical significance. The MR Egger method did not provide evidence for a causal effect in either analysis. Heterogeneity statistics did not indicate any significant evidence of heterogeneity, and the causal direction test strongly supported the hypothesis that CLIC5 caused HT. Horizontal pleiotropy was not detected. The consistency of the findings across different outcome databases enhances the credibility of the observed causal effect between CLIC5 and HT. Although the effect estimates varied slightly, these variations can be attributed to differences in the underlying genetic architectures captured by the respective GWAS studies.

Background Clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer with a high metastatic rate and high mortality rate. The molecular mechanism of ccRCC development, however, needs further study. Aurora kinase B (AURKB) functions as an important oncogene in various tumors; therefore, in the present study, we aimed to explore the mechanism by which AURKB affects ccRCC development. Methods We performed bioinformatics analysis, CCK-8 assay, RNA sequencing, RT-PCR and Western blot to analyze the function and mechanism of AURKB in ccRCC. Results TIMER2.0 showed that AURKB was overexpressed in Kidney Renal Clear Cell Carcinoma (KIRC), the UALCAN database showed the survival rate of KIRC patients with different expression levels of AURKB and different gender indicated in the same gender, high AURKB expression predicts lower survival rate. Silencing of AURKB expression inhibits the proliferation of ccRCC cells. RNA-seq data suggested that AURKB is involved in fatty acid metabolism. Silencing of AURKB inhibited the expression of fatty acid synthase (FASN). FASN is a key gene involved in fatty acid metabolism. TIMER2.0 showed that FASN is upregulated in KIRC. Silencing of FASN inhibited the proliferation of ccRCC cells. Conclusions AURKB induces the proliferation of ccRCC cells by regulating fatty acid metabolism.

Background: There is growing evidence that the gene named tumor necrosis factor α–induced protein 6 (TNFAIP6) has an important role in various tumors. However, a systematic pan-cancer analysis of TNFAIP6 is lacking. Here we aimed to analyze the expression of TNFAIP6 across multiple cancers and verify its expression during the progression of colon cancer. Methods: We performed a comprehensive bioinformatics analysis to examine the expression of TNFAIP6 across 27 tumor types. GEPIA2 was used to evaluate the effect of TNFAIP6 on clinical cancer prognosis. cBioportal was used to assess TNFAIP6 mutations. The correlation between TNFAIP6 and cancer immune infiltrates was explored using TIMER2.0. The CancerSEA database was used to perform functional analysis of TNFAIP6. Metascape was used to identify TNFAIP6-related gene enrichment pathways. Immunohistochemistry was performed to detect TNFAIP6 protein expression in the colon cancer. In addition, the Comparative Toxicogenomics Database was searched for known and possible antitumor drugs that may be associated with TNFAIP6. Results: We found that, in most of the cancers included in this analysis, TNFAIP6 was highly expressed, and there is a distinct relationship between TNFAIP6 expression and cancer prognosis. TNFAIP6 expression is associated with cancer-associated fibroblasts, neutrophils, and endothelial cells. TNFAIP6 and similar genes may also be involved in the PID_VEGF_VEGFR_ pathway. Immunohistochemistry revealed an increasing trend of TNFAIP6 protein expression in normal, adenoma, and colon cancer tissues. Several known and possible antitumor drugs that may be associated with TNFAIP6 were identified in the Comparative Toxicogenomics Database. These results suggest that a number of drugs may target TNFAIP6 during cancer treatment, including cisplatin, irinotecan, resveratrol, U 0126, NSC689534, genistein, NSC668394, oxaliplatin, plerixafor, topotecan, vincristine, flutamide, doxorubicin, MRK 003, folic acid, demecolcine, tunicamycin, zoledronic acid, and schizandrin B. Conclusions: TNFAIP6 may function as an oncogene in certain cancers. Furthermore, this study provides evidence that TNFAIP6 is an important factor in colon cancer progression.

Introduction: Colorectal cancer (CRC) is a serious threat to human health. Screening new biomarkers can provide basis for improving the prognosis and individualized treatment of CRC. Although some members of the defensin family were found increased in pancreatic cancer and CRC, their exact function and clinical significance remain unclear.Methods: In this study, the expression, correlation, mutation, and functional enrichment of several defensin family members in pancreatic cancer and CRC were analyzed using tumor public databases and verified in several patients.Results: Results showed no significant correlation between the expression levels of DEFA1-4 and CRC. The expression levels of DEFA5 and DEFA6 significantly increased in CRC tissues compared with those in normal tissues. DEFA5 may be associated with better prognosis of CRC, while DEFA6 may be associated with poor prognosis. Immunohistochemistry (IHC) experiments showed that the expression of DEFA6 was significantly higher in adenoma than in normal mucosa and slightly higher in carcinoma than in normal mucosa. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis found that DEFAs were closely related to hsa05202: transcriptional misregulation in cancer and Hsa04015: Rap1 signaling pathway. DEFA5 may be a stable and good prognostic marker, and DEFA6 may be a poor prognostic marker in CRC of metastasis.Conclusion: Overall, DEFA5 and DEFA6 have a certain degree of sensitivity and specificity in predicting CRC.

Background. Skin cutaneous melanoma is one of most aggressive type of cancers worldwide. Therefore, the identification of SKCM biomarkers is of great importance. FLG gene is one of the genes that encode proteins involved in epidermal formation. This was the first time to study the role of FLG in the prognosis and immune infiltrates of skin cutaneous melanoma. Methods. We downloaded the somatic mutation data of 471 SKCM patients from the Cancer Genome Atlas (TCGA) database and analyzed the mutation profiles with MafTools package. The expression of FLG and the overall survival in SKCM were analyzed by GEPIA. Additionally, univariate and multivariate Cox analyses were used to compare several clinical features with survival rates. We used TIMER to investigate FLG expression and collection of immune infiltration levels in SKCM, as well as cumulative survival in SKCM. Meanwhile, we also used CIBERSORT to investigate the association between FLG and cancer immune infiltration. In addition, gene set enrichment analysis (GSEA) was performed using the TCGA dataset. Furthermore, data from GEO and HPA was used to validate the results. Results. Single nucleotide polymorphism (SNP) happened more frequently than insertion or deletion, and CT was the most common of SNV in SKCM. We selected the first 15 mutated genes by analyzing 471 melanoma samples, and the prognosis analysis showed that only the high expression of mutated FLG gene was significantly correlated with the poor prognosis of SKCM. Multivariate Cox analysis showed that age, the worse tumor status, less lymph node metastasis, and FLG expression were independent factors for prognosis. Specifically, lower infiltration levels of B cell, CD8+ T cells, neutrophils, and dendritic cells correlated with poor survival outcomes in SKCM. GSEA revealed that FLG is closely related to cancer pathways and epidermal cell proliferation. In addition, the previous conclusions can be verified from external data from GEO and HPA. Conclusion. The discovery of mutant gene FLG as a biomarker of SKCM helps elucidate how changes in the immune environment promote the occurrence of cutaneous melanoma. Further analysis suggested that FLG might be a new predictor of SKCM prognosis.