Molecular pathological mechanism of DIQUAT-induced multi-organ injury

Diquat is one of the most commonly used pesticides in recent years, and its improper use can lead to acute poisoning and loss of life. This study employed single-cell and single-cell nucleus transcriptome sequencing on mouse lung, liver, and kidney tissues induced by diquat to unveil the molecular pathophysiology of multi-organ damage caused by diquat exposure. In this research, we emphasize a novel hepatic cell subpopulation associated with detoxification and metabolic dysregulation following diquat exposure in the liver. Furthermore, a robust and intricate immune-inflammatory response is a prominent feature in the lungs, while a time-dependent shift from glomerular infiltration to tubular injury pattern is the main characteristic in the kidneys. This study underscores distinct injury features in multiple organs, significantly enhancing our understanding of diquat-induced multi-organ damage. Relevant molecules and pathways identified may serve as potential novel molecular targets. Harvested cells at the required densities were combined with gel beads containing the barcoded information along with a mixture of cells and enzymes. Oil surfactant droplets in a microfluidic double-cross system were used for encapsulation to generate Gel Beads-In-Emulsions (GEMs). GEMs were passed through a reservoir and collected while the gel beads were lysed to release the barcode sequence. The cDNA fragment was then reverse transcribed before the sample was labeled. The gel beads and the oil droplets are ruptured and PCR amplification was performed using the generated cDNA as a template. The products of all GEMs were mixed and a standard sequencing library is constructed. The cDNA was first enzymatically digested into fragments of about 200300 base pairs (bp), together with the library building process of traditional secondgeneration sequencing such as sequencing junction and primers. Finally, the DNA library was generated by PCR amplification