Hypothesis Trypanosomiasis is one of the main threats to human and animal health in African countries. Trypanosoma brucei can evade the host immune recognition by rapidly altering its variant surface glycoprotein (VSG). The ATP synthase F 1 subunit of the parasite exhibits extremely low similarity to that of its mammalian hosts, hypothetically making it an ideal target for the development of novel therapeutics. Experiments Graphene quantum dots (GQDs) were synthesized, and their adhesion to T. brucei surface and internalization was observed microscopically. The activity of ATP synthase and mitochondrial membrane potential of T. brucei were measured after exposure to GQDs. Proteomics, biolayer interferometry, and molecular dynamic simulations were utilized to evaluate the interaction between GQDs with the target proteins. Findings GQDs specifically adhered to the VSG of T. brucei and were conveyed inside the parasite via the VSG internalization pathway. The GQDs promoted intracellular ROS production, interacted with, and inhibited the activity of the p18 subunit of ATP synthase, disrupted parasite mitochondrial membrane potential. Additionally, the GQDs caused a decrease in aminoacyl – tRNA biosynthesis, and upregulated RNA and protein degradation pathways. The findings of this study offer a novel avenue for the target-oriented discovery of anti-trypanosome drugs.
Isolation of high-quality DNA is one of the essential steps in the field of molecular biology. Before DNA isolation, ribonuclease A (RNase) has to be introduced to degrade RNA. However, RNase needs to be stored at low temperature, and the conditions for the degradation of RNA need to be strictly controlled. In the study, graphene oxide functionalized magnetic particles (GO-MPs) were prepared and used for RNA removal. GO-MPs demonstrate good environmental stability. The RNA removing process by GO-MPs can be finished in 10 min at room temperature. Also, GO-MPs exhibit good selectivity toward RNA over double-stranded DNA. Next, the spherical and core–shell structure poly(3,4-dihydroxy-L-phenylalanine) functionalized MPs (polyDOPA-MPs) were prepared and used for the purification of DNA. The DNA adsorption and release can be realized by adjusting the pH value of solutions. It is worth emphasizing that the condition for RNA removal by GO-MPs is the same as that for DNA adsorption by polyDOPA-MPs, which ensures the effective connection between the two steps. The feasibility of the MPs-based protocol was verified by the successful extraction of genomic DNA from Escherichia coli DH5α and genetically modified corn. The results suggest that the protocol would provide a promising platform for RNase-free RNA removal and DNA purification from bacteria and plant seeds.
Di-(2-ethylhexyl) phthalate (DEHP) is an environmental endocrine disruptor that accumulates in organisms in various ways and induces male reproductive system disorders. In this study, we established a testicular injury model by gavage with different concentrations of DEHP. The testes were then collected for RNA sequencing (RNA-seq), and the results were analyzed by bioinformatics and verified by experiments. Our research results show that different concentrations of DEHP interfere with testicular development differently. Weighted gene coexpression network analysis (WGCNA) generated sixteen modules and identified the turquoise module as key. Then, estrogen receptor 1 (ESR1), filamin A (Flna) and Furin were identified as hub genes. qPCR and immunohistochemistry results revealed that all three hub genes were upregulated. We detected the locations of these genes by immunohistochemistry. ESR1 was mainly located in Leydig cells; Flna immunostaining is observed in the Leydig and some germ cells and Furin staining was seen in almost all types of testicular cells. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed enrichment mainly in MAPK signaling pathways, p53 signaling pathways, HIF-1 signaling pathways, protein processing in the endoplasmic reticulum, apoptosis, the cell cycle, RNA degradation, etc. This is the first study using WGCNA to investigate the mechanism of DEHP-induced injury in the prepubertal testis, providing new research angles to further understand the mechanism of DEHP-induced injury in the prepubertal testis.
危险品化学品经营许可证（带存储）