ABSTRACT
Objective To explore structural differences of intestinal flora in primary insomnia patients with different TCM syndromes through the high-throughput 16S rDNA sequencing analysis. Methods Totally 65 patients with primary insomnia were divided into 22 patients with syndrome of liver depression transforming into fire, 17 patients with deficiency of both heart and spleen syndrome, 26 patients with syndrome of hyperactivity of fire due to yin deficiency, with 47 cases of healthy people as the control group. The fecal flora structure of the subjects was analyzed by high-throughput 16S rDNA sequencing. QIIME software and R language stats package were used to analyze the diversity of flora. Results Totally 1226 different operational taxonomic units (OUTs) were obtained, and there were 180 significant differences among the 4 groups (P<0.05), indicating that the samples were rich in microbial colonies. The mapped reads in group of liver depression transforming into fire and hyperactivity of fire due to yin deficiency were more than the group of deficiency of both heart and spleen and the control group (P<0.05). Unweighted UniFrac analysis showed that the difference among groups was remarkably greater than the difference within group, and the grouping was statistically significant (R=0.103, P=0.002). It suggested that the diversity of intestinal flora was highly correlated with different TCM syndromes of insomnia. There were a total of 57 genera found significant differences among the different groups at the genus level (P<0.05), and 115 species at all species level. The dominant flora of the control group were prevotella, megamonas, clostridium Ⅺ (clostridium ⅩⅧ), weissella, and alloprevotella; The dominant flora of liver depression transforming into fire syndrome were phascolarctobacterium, flavonifractor, eggerthella, and bilophila; The dominant flora of deficiency of both heart and spleen syndrome were sphingomonas and methylobacterium; The dominant flora in hyperactivity of fire due to yin deficiency syndrome group were bacteroides, parabacteroides, parasutterella, butyricimonas, odoribacter. Conclusion The patients with primary insomnia have abundant intestinal flora diversity and diverse flora structure, which may affect the occurrence, development and outcome of different TCM syndromes.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the methylation status in promoter region of norepinephrine transporter gene (NET, SLC6A2) in heart failure ( HF) patients and its correlation with qi deficiency/blood stasis syndrome (QDS/BSS).</p><p><b>METHODS</b>Thirty-six patients with heart failure (NYHA classification III to IV) were recruited in the study (as the heart failure group) and their scores of QDS/BSS were evaluated. Besides, a healthy elderly group (30 cases) and a healthy youth group (30 cases) were also set up. They were recruited from Physical Examination Center of Fujian Provincial Hospital. Pyrosequencing was applied to detect the methylation in promoter region of SLC6A2 gene, and the total methylation index (MTI) of CpG island was calculated. The correlation between the methylation status in promoter region of SLC6A2 and scores of QDS/BSS was assessed using Pearson and Partial analyses. Risk factors were screened and adjusted using Logistic regression.</p><p><b>RESULTS</b>By one-factor analysis of variance, the total MTI in the HF group (219.72% ± 54.03%) was obviously higher than that in the healthy elderly group (194.47% ± 34.92%) and the healthy youth group (161.60% ± 41.11%) (all P < 0.05). Meanwhile, the total MTI was higher in the healthy elderly group than in the healthy youth group (P < 0.01). By covariance analysis , after controlling age and BMI, the total MTI was higher in the HF group than in the healthy elderly group (P = 0.041), while it was higher in the healthy elderly group than in the healthy youth group (P = 0.016). Age was found to play an essential role in affecting MTI of SLC6A2 gene promoter region among the 3 groups (F = 16.447, P = 0.01). The total MTI was quite lower in the healthy youth group. Results of Partial correlation analysis showed MTI was positively correlated with scores of qi deficiency and blood stasis respectively (r = 0.494 and 0.419 respectively, both P < 0.05). Logistic regression analysis showed after adjusting confounding factors, the relative risk (OR value) of total MTI of SLC6A2 gene in promoter region was 1.038 (95% CI, 1.006 to 1.071, P = 0.020).</p><p><b>CONCLUSIONS</b>Abnormally elevated methylation of the promoter region of SLC6A2 gene is one of risk factors for HF. In addition, the degree of methylation of the promoter region of SLC6A2 gene was positively correlated with the severity of QDS/BSS.</p>