The WRKY web, which is comprised of a subset of WRKY transcription factors (TFs), plays a crucial role in the regulation of plant immunity, however, the mode of organization and operation of this network remains obscure, especially in non-model plants such as pepper (Capsicum annuum). Herein, CaWRKY22, a member of a subgroup of IIe WRKY proteins from pepper, was functionally characterized in pepper immunity against Ralstonia Solanacearum. CaWRKY22 was found to target the nuclei, and its transcript level was significantly upregulated by Ralstonia Solanacearum inoculation (RSI) and exogenously applied salicylic acid (SA), Methyl jasmonate (MeJA), or ethephon (ETH). Loss-of-function CaWRKY22, caused by virus-induced gene silencing (VIGS), enhanced pepper’s susceptibility to RSI. In addition, the silencing of CaWRKY22 perturbed the hypersensitive response (HR)-like cell death elicited by RSI and downregulated defense-related genes including CaPO2, CaPR4, CaACC, CaBPR1, CaDEF1, CaHIR1, and CaWRKY40. CaWRKY22 was found to directly bind to the promoters of CaPR1, CaDEF1, and CaWRKY40 by chromatin immuno-precipitation (ChIP) analysis. Contrastingly, transient overexpression of CaWRKY22 in pepper leaves triggered significant HR-like cell death and upregulated the tested immunity associated maker genes. Moreover, the transient overexpression of CaWRKY22 upregulated the expression of CaWRKY6 and CaWRKY27 while it downregulated of the expression of CaWRKY58. Conversely, the transient overexpression of CaWRKY6, CaWRKY27, and CaWRKY40 upregulated the expression of CaWRKY22, while transient overexpression of CaWRKY58 downregulated the transcript levels of CaWRKY22. These data collectively recommend the role of CaWRKY22 as a positive regulator of pepper immunity against R. Solanacearum, which is regulated by signaling synergistically mediated by SA, jasmonic acid (JA), and ethylene (ET), integrating into WRKY networks with WRKY TFs including CaWRKY6, CaWRKY27, CaWRKY40, and CaWRKY58.

Background The modification 6‐methyladenine (m6A) is the most common type in RNA methylation. Our study aims to explore the bioinformatic analysis of m6A in endometrial cancer. Methods The expression of 23 m6A RNA methylation regulators was compared through The Cancer Genome Atlas (TCGA) database among 406 endometrial tissue and 19 normal tissue samples. The Wilcoxon test was applied to compare the relationship between the clinicopathological characteristics and expression. Cox regressions were performed to identify the prognostic factors associated with overall survival. Gene ontology (GO) and Gene Set Enrichment Analysis (GSEA) were performed to evaluate the potential pathways. Results YTHDF2, HNRNPA2B1, HDRNPA2B1, YTHDF1, FMR1, IGF2BP3, METTL13, RBM15B, IGF2BP1, YTHDF3, YTHDC1, ZC3H13 IGF2BP2, KIAA1429, METTL14, RBMX, FTO, ALKBH5, and METTL16 were significantly abnormally expressed in endometrial cancer tissue samples. Both univariate and multivariate Cox regression analyses indicated that age, grade, and risk score were independent risk factors. High expression of FTO was associated with worse overall survival. Conclusion M6A RNA methylation regulators play vital role in endometrial cancer.

Cervical cancer is one of the most common gynecological malignancies. Due to the high heterogeneity of cervical cancer accelerating cancer progression, it is necessary to identify new prognostic markers and treatment regimens for cervical cancer to improve patients’ survival rates. We purpose to construct and verify a risk prediction model for cervical cancer patients. Based on the analysis of data from the Gene Expression Omnibus database (GEO) and The Cancer Genome Atlas (TCGA), differences of genes in normal and cancer samples were analyzed and then used analysis of WGCNA along with consistent clustering to construct single-factor + multi-factor risk models. After regression analysis, the target genes were obtained as prognostic genes and prognostic risk models were constructed, and the validity of the risk model was confirmed using the receiver operating characteristic curve (ROC) and Kaplan–Meier curve. Subsequently, the above model was verified on the GSE44001 data validation followed by independent prognostic analysis. Enrichment analysis was conducted by grouping the high and low risks of the model. In addition, differences in immune analysis (immune infiltration, immunotherapy), drug sensitivity, and other levels were counted by the high and low risks groups. In our study, three prognostic genes including APOD, APOC1, and SQLE were obtained, and a risk model was constructed along with validation based on the above-mentioned analysis. According to the model, immune correlation and immunotherapy analyses were carried out, which will provide a theoretical basis and reference value for the exploration and treatment of cervical cancer.

ObjectiveTo investigate the clinicopathological characteristics and overall survival in high-risk human papillomavirus (HPV)-independent and HPV-associated cervical cancer.MethodsPatients with cervical cancer hospitalized between September 2015 and December 2019 from the Affiliated Cancer Hospital of Guizhou Medical University were enrolled. First, patients with negative results by HPV primary screening were excluded. Second, the paraffin-embedded tumor tissues from patients with negative results were used for extraction of deoxyribonucleic acid (DNA). The Hybribio K-37 test (PCR and flow-through hybridization for 37 types of HPV) was used to further identify HPV-negative infection status. Finally, 1:4 propensity score matching between high-risk HPV-independent and HPV-associated groups was performed, and the clinicopathological characteristics and overall survival were compared between the two groups.ResultsForty cervical HPV primary screening negative patients were screened of 729 patients (5.5%). Among them, 13 (1.8%) patients who were identified with high-risk HPV-independent cervical cancer after the K-37 test were selected as the study group. There were significant intergroup differences in the distribution of International Federation of Gynecology and Obstetrics (FIGO, 2018) stage (χ2=5.825, p=0.016), pathological types (χ2=6.910, p=0.009), lymph node metastasis (χ2=6.168, p=0.013), and tumor size (χ2=5.319, p=0.021). After propensity score matching, 52 patients from the HPV-associated group were selected as the control group. Patients with high-risk HPV-independent cervical cancer had poorer prognosis than those with HPV-associated cervical cancer (median overall survival: 27 vs 29 months, p=0.03; median disease-free survival: 27 vs 29 months, p=0.021).ConclusionPatients with high-risk HPV-independent cervical cancer more frequently had advanced stage disease, nodal metastasis, larger tumor, and a higher proportion of adenocarcinoma. The prognosis of patients with high-risk HPV-independent cervical cancer was poorer than those with HPV-associated cervical cancer.