Gut microbiota signatures in patients with minimal hepatic encephalopathy due to HBV-associated cirrhosis

Sleep disturbances severely deteriorate the quality of life in cirrhotic patients with minimal hepatic encephalopathy (MHE). In this study, ninety patients with MHE due to HBV-associated cirrhosis were randomly assigned to the exploratory (n=60) and validation (n=30) cohorts. The gut microbiome was analysed by bacterial 16S rRNA sequencing. Specifically, the V3-V4 hypervariable regions of bacterial 16S ribosomal RNA (rRNA) genes were amplified using the primer sequences: forward 341F (5'- CCTACGGGNGGCWGCAG-3') and reverse 806R (5'-GGACTACHVGGTATCTAAT-3'). After amplification, the amplicons were purified, quantified, and subjected to library preparation for sequencing. The prepared library was sequenced on the MiSeq PE300 platform (Illumina, San Diego, CA, USA) at Majorbio Bio-Pharm Technology Co. Ltd. (Shanghai, China). This platform employs a paired-end sequencing approach with read lengths of up to 300 nucleotides for each fragment end.