Hiseq platform

The miR-21 expression data and the clinical data of the LGG patients and the GBM patients were obtained from the TCGA database. MiR-21 expression was measured by Illumina Hi-Seq platform in the LGG dataset and Agilent 8 × 15 K Human microRNA platform in the GBM dataset. The inclusion criteria for the patients in the LGG and GBM datasets were as follows: (a) miR-21 expression level data and the corresponding follow-up data were available; (b) the OS or PFS were ≥ 30 days; and (c) clinical data of the patients, such as age at diagnosis, gender, tumor grade, and KPS, were available. The high- and low-expression groups were distinguished by the median value of the miR-21 expressions. Unpaired t test and chi-square test were used for the comparison of high miR-21 expression group and low miR-21 expression group. The survival curves were estimated by the Kaplan–Meier method. Survival differences between the high-expression group and the low-expression group were assessed by a log-rank test. A multivariate Cox regression analysis was used to identify miR-21 expression as an independent prognostic biomarker. All the statistical analyses were performed using PASW Statistics Version 18.0 (SPSS Inc., Chicago, Illinois, USA).

The Beijing Genomics Institute (BGI) performed the 16S rRNA gene sequencing. DNA from fecal pellets collected from APL patience was amplified using primer 515F (5′-GTGCCAGCMGCCGCGGTAA-3′) and 806R (5′-GGACTACHVGGGTWTCTAAT-3′) to target the V4 regions of 16S rRNA of bacteria. PCR reaction was carried out with 30 ng DNA samples and primers. The PCR amplification products were purified by Agencourt AMPure XP magnetic beads, and labeled, and then library building was completed. The range and concentration of fragments in the library and qualified libraries sequencing based on inserted fragment size were measured by Agilent 2100 Bioanalyzer and HiSeq platform. The 16s sequencing data were deposited in NCBI with Submission ID: SUB12873522, BioProject ID: PRJNA935705 (Raw sequence reads project Accession: PRJNA935705) with link https://www.ncbi.nlm.nih.gov/bioproject/?term=PRJNA935705 (accessed on 27 February 2023).