The biological mechanisms related to hub genes in the development of heart failure in dialysis patients were investigated through transcriptome sequencing. The biological mechanisms related to hub genes in the development of heart failure in dialysis patients were investigated through transcriptome sequencing

Purpose: Heart failure (HF) has a very high prevalence in patients with maintenance hemodialysis (MHD). However, there is still a lack of effective and reliable HF diagnostic markers and therapeutic targets for patients with MHD. Patients and methods: In this study, we analyzed transcriptome profiles of 28 patients with MHD by high-throughput sequencing. Overall design: Sample preparation and processing Peripheral blood mononuclear cell (PBMC) samples of 15 dialysis patients without heart failure (Normal) and 15 dialysis patients with heart failure (Case) human were collected for mRNA transcriptome sequencing. RNA was isolated and purified from the total samples using TRlzol (invitrogen, CA,USA) according to the manufacturer's protocol. NanoDrop ND-1000 (Wilminton, DE, USA) was used to control the total RNA sender bulk and cleanliness. The fragmented RNA was synthesized into cDNA by reverse transcriptase, and the cDNA was amplified and purified. Building the library and prepared for sequencing on the illumina Novaseq 6000 sequencing platform. After the inferior-quality reads were deleted, the cleanreads were aligned into the human genomic reference (GRCh38_gencode_v33) by HISATI16.

研究背景：心力衰竭（Heart Failure, HF）在维持性血液透析（Maintenance Hemodialysis, MHD）患者中患病率极高，但目前仍缺乏针对MHD患者的有效且可靠的HF诊断标志物与治疗靶点。对象与方法：本研究通过高通量测序技术分析了28例MHD患者的转录组表达谱。总体实验设计：样本制备与处理 收集15例无心力衰竭的透析患者（正常组）及15例伴心力衰竭的透析患者（病例组）的人类外周血单个核细胞（Peripheral Blood Mononuclear Cell, PBMC）样本，开展mRNA转录组测序。按照试剂制造商的操作规程，使用TRlzol（美国加利福尼亚州Invitrogen公司）从总样本中分离并纯化RNA。采用NanoDrop ND-1000（美国特拉华州威尔明顿市）检测总RNA的总量与纯度。将片段化的RNA经逆转录酶反转录合成cDNA，随后对cDNA进行扩增与纯化。构建测序文库后，在Illumina NovaSeq 6000测序平台上完成测序。去除低质量读数后，通过HISATI16将清洁读数（cleanreads）比对至人类基因组参考序列（GRCh38_gencode_v33）。