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Background: Breast cancer (BC) is the leading malignancy among women worldwide. Aim: This work aimed to present a comprehensively bioinformatic analysis of gene expression profiles and to identify the hub genes during BC tumorigenesis, providing potential biomarkers and targets for the diagnosis and therapy of BC. Materials & Methods: In this study, multiple public databases, bioinformatics approaches, and online analytical tools were employed and the real‐time reverse transcription polymerase chain reaction was implemented. Results: First, we identified 10, 107, and 3869 differentially expressed genes (DEGs) from three gene expression datasets (GSE9574, GSE15852, and GSE42568, covering normal, para‐cancerous, and BC samples, respectively), and investigated different biological functions and pathways involved. Then, we screened out 8, 16, and 29 module genes from these DEGs, respectively. Next, 10 candidate genes were determined through expression and survival analyses. We noted that seven candidate genes JUN, FOS, FOSB, EGR1, ZFP36, CFD, and PPARG were downregulated in BC compared to normal tissues and lower expressed in aggressive types of BC (basal, HER2+, and luminal B), TP53 mutation group, younger patients, higher stage BC, and lymph node metastasis BC, while CD27, PSMB9, and SELL were upregulated. The present study discovered that the expression levels of these candidate genes were correlated with the infiltration of immune cells (CD8+ T cell, macrophage, natural killer [NK] cell, and cancer‐associated fibroblast) in BC, as well as biomarkers of immune cells and immune checkpoints. We also revealed that promoter methylation, amplification, and deep deletion might contribute to the abnormal expressions of candidate genes. Moreover, we illustrated downstream‐targeted genes of JUN, FOS, FOSB, EGR1, and ZFP36 and demonstrated that these targeted genes were involved in "positive regulation of cell death", "pathways in cancer", "PI3K‐Akt signaling pathway", and so on. Discussion & Conclusion: We presented differential gene expression profiles among normal, para‐cancerous, and BC tissues and further identified candidate genes that might contribute to tumorigenesis and progression of BC, as potential diagnostic and prognostic targets for BC patients. [ABSTRACT FROM AUTHOR]

A simple and accurate PCR protocol for identification of Fusarium graminearum from fungal cultures was developed. In this protocol, DNA extraction was omitted by using the Thermo Scientific Phire direct PCR master mix kit for template preparation and PCR. Two primer pairs, ITS5/ITS2 for DNA template quality control and Fg16F/Fg16R for F. graminearum identification, were used in a duplex PCR. Each 20-µL PCR contained 10 µL 2× Phire Direct PCR master mix, 2 µL template and 0.25 µM each of the four primers. The PCR program consisted of an initial denaturation at 98°C for 5 min, followed by 40 cycles of denaturation at 98°C for 5 s, annealing at 62°C for 5 s and extension at 72°C for 20 s, and a final extension at 72°C for 1 min. This protocol was used to test 196 fungal cultures isolated from 75 wheat samples. Fourteen cultures were identified as F. graminearum. This protocol was demonstrated to be simpler and more accurate than other PCR protocols for F. graminearum identification. [ABSTRACT FROM AUTHOR]