Analysis of the glutathione S-transferase (GST) gene expression in an insecticide-resistant strain of Cydia pomonella using real-time quantitative polymerase chain reaction is a key step toward more mechanism studies that require suitable reference genes with stable expression. Here, nine commonly used reference genes were selected, and their expression stabilities were analyzed. Results showed that EF-1α was the most stable reference gene in all of the experimental sets. The combinations of EF-1α and 18S, EF-1α and RPL12, and EF-1α and GAPDH were sufficient for normalization of gene expression analysis accurately in developmental stages, tissues, and larvae exposed to sublethal dose of λ-cyhalothrin, respectively. Additionally, the suitability of particular reference genes was verified by analyzing the spatiotemporal and insecticide-induced expression profiles of CpGSTe3, CpGSTd3, and CpGSTd4, which were overexpressed in a λ-cyhalothrin-resistant population from northeast China. These genes were used to confer the practicability of reference genes chosen in this study.
Over two hundred health awareness events take place in the United States in order to educate the public about various diseases. It would be informative and instructive for the organizations to know the impact of these events, although such information could be difficult to measure. We investigated whether 46 selected events attract the public attention by increasing the search frequencies of certain keywords. Internet search data from 2004 to 2017 were downloaded from Google Trend (GT). Three statistical methods including Transfer Function Noise modeling, Wilcoxon Rank Sum test, and Binomial inference were conducted. Our study showed that 10 health awareness events resulted in increased search frequencies in the event months, and 28 events did not, with the rest being classified as unclear.
In 2006, Takahashi and Yamanaka first created induced pluripotent stem cells from mouse fibroblasts via the retroviral introduction of genes encoding the transcription factors Oct3/4, Sox2, Klf44, and c-Myc. Since then, the future clinical application of somatic cell reprogramming technology has become an attractive research topic in the field of regenerative medicine. Of note, considerable interest has been placed in circumventing ethical issues linked to embryonic stem cell research. However, tumorigenicity, immunogenicity, and heterogeneity may hamper attempts to deploy this technology therapeutically. This review highlights the progress aimed at reducing induced pluripotent stem cells tumorigenicity risk and how to assess the safety of induced pluripotent stem cells cell therapy product.
Alzheimer's disease (AD) is a global health issue, but the precise underlying mechanism has not yet been elucidated. The present study aimed to integrate microRNA (miRNA or miR) and mRNA profiles of AD and identify hub genes via bioinformatics analysis. Datasets associated with AD (GSE113141, GSE104249 and GSE138382) were integrated. Bioinformatics analysis was used to identify the hub mRNAs. TargetScan was used to predict miRNAs that have binding sites for the hub genes. Reverse transcription-quantitative (RT-q)PCR and western blot analysis was performed to assess miRNA and mRNA expression levels in APP/PS1 transgenic mice and human U251 cells. Luciferase reporter assay and RNA interference were utilized to verify the functions of these miRNAs in vitro. Bioinformatics analysis demonstrated that expression levels of the gene encoding transmembrane immune signaling adaptor TYROBP were upregulated in both the GSE113141 and GSE104249 datasets; TYROBP also served as the hub gene in AD. miR-628-5p was predicted to have binding sites for TYROBP and was downregulated in GSE138382. RT-qPCR confirmed low miR-628-5p and high TYROBP expression levels in APP/PS1 transgenic mice and human U251 cells. Western blot analysis demonstrated high protein expression levels of amyloid β (Aβ) precursor protein, Aβ and TYROBP in APP/PS1 transgenic mice and U251 cells. Dual luciferase reporter assay confirmed that TYROBP was targeted by miR-628-5p. miR-628-5p/TYROBP may inhibit progressive neurodegeneration in AD and could be used as novel biomarkers and candidate drug targets.
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