Study on potential biomarkers of energy metabolism‐related to early‐stage Yin‐deficiency‐heat syndrome based on metabolomics and transcriptomics

Abstract

Yin‐deficiency‐heat (YDH) syndrome is a common sub‐health state of the human body in traditional Chinese medicine (TCM). However, due to the lack of objective quantitative diagnostic indicators, patients with early‐stage YDH syndrome cannot be treated in time and can develop a pathological (disease) state. Therefore, it is necessary to apply modern diagnostic techniques in order to identify the biological markers for the diagnosis of early‐stage YDH syndrome. In the present study, we performed Solexa sequencing and non‐targeted metabolomics analysis using high‐performance liquid chromatography coupled with mass spectrometry to screen differentially expressed mRNAs and differential metabolites in individuals with early‐stage YDH syndrome and healthy controls. Bioinformatics methods were used to perform enrichment analysis of differentially expressed mRNAs and differential metabolites for biological functions and signaling pathways. Furthermore, we found that differentially expressed mRNAs and differential metabolites were related to energy metabolism. Real‐time PCR was used to validate the mRNA expression in the serum of subjects with early‐stage YDH syndrome. We found that the mitochondrially encoded NADH dehydrogenase 2 (MT‐ND2) mRNA was differentially expressed in the serum of individuals with early‐stage YDH syndrome. Receiver operating characteristic (ROC) curve and logistic regression analysis were used to evaluate the efficacy of the diagnostic model based on eight differential metabolites. We combined the three metabolites such as Glycine, Sphingomyelin, and Isocitrate to establish the diagnostic model with a sensitivity of 0.853 and a specificity of 0.800. The combination of the above three metabolites may serve as a potential biomarker for the diagnosis of early‐stage YDH syndrome. Our study reveals potential biomarker for the diagnosis of early‐stage YDH syndrome and also provides a new method for the quantification and objectification of TCM syndromes.

Transcriptomic and metabolomic analyses reveal mechanisms of adaptation to salinity in which carbon and nitrogen metabolism is altered in sugar beet roots

“…After the total RNA was quantified, the eukaryotic mRNA was enriched by attachment to Oligo (dT) magnetic beads. The extracted mRNA was randomly broken into short fragments by fragmentation buffer, and the fragmented mRNA was used as a template to synthesize a strand of cDNA with six-base random primers (random hexamers), double-stranded cDNA synthesis in buffer, dNTPs, RNaseH and DNA Polymerase I [94]. AMPure XP beads were used to purify the double-stranded product, both T4 DNA polymerase and Klenow DNA polymerase were used to repair attachment of the sticky end of the DNA to the blunt end, A bases and linkers were added to the 3′ end, AMPureXP beads were used for fragment selection, and PCR amplification was ultimately performed to increase the final sequencing library.…”

“…The expression levels of all transcripts were calculated by StringTie and Ballgown. StringTie was also used to determine mRNA expression levels by calculating the fragments per kilobase of transcript per million mapped reads (FPKM) [94]. Differentially expressed mRNAs and genes were selected by the R package Ballgown according to the criteria of log 2 (fold change) > 1 or log 2 (fold change) < − 1 and were statistically significant when P < 0.05 [95].…”

The Anatomical Record continues to replace wonder with knowledge through first‐hand experience of acupuncture

“…The second and third papers focus on potential biomarkers for the same TCM syndrome (Chen et al, 2020; Yi et al, 2020). The fourth and fifth papers identify potential biomarkers, using metabolomics and transcriptomics approaches, for a related TCM syndrome, Yin ‐deficiency syndrome (Gan et al, 2020; Yu et al, 2020). Next in the lineup is a paper on different TCM syndromes that are related to multidrug‐resistant tuberculosis (Hu et al, 2020).…”

Study on the Effect of Macrophages on Vascular Endothelium in Mice With Different TCM Syndromes of Dyslipidemia and its Biological Basis Based on RNA-Seq Technology

“…Transcriptomics is a technique used to study gene transcription and regulation related to TCM syndromes and is conducive to in-depth biological research toward a better understanding of the pathogenesis of TCM syndromes, and so warrants more prevalent application in TCM syndrome research ( Jiang and Li, 2020 ). Gan et al (2020) studied Yin deficiency–heat syndrome using transcriptomics technology and obtained its potential biomarkers. Currently, though, studies concerning the characteristics of SKYD syndrome and PDR syndrome in relation to dyslipidemia are rarely reported.…”