[Methylation in Promoter Region of SLC6A2 Gene in Heart Failure Patients and Its Correlation with Qi Deficiency/Blood Stasis Syndrome].

OBJECTIVE: 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). METHODS: 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. RESULTS: 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). CONCLUSIONS: 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.

An investigation of the antidepressant action of xiaoyaosan in rats using ultra performance liquid chromatography-mass spectrometry combined with metabonomics.

A rapid, highly sensitive, and selective method was applied in a non-invasive way to investigate the antidepressant action of Xiaoyaosan (XYS) using ultra performance liquid chromatography-mass spectrometry (UPLC-MS) and chemometrics. Many significantly altered metabolites were used to explain the mechanism. Venlafaxine HCl and fluoxetine HCl were used as chemical positive control drugs with a relatively clear mechanism of action to evaluate the efficiency and to predict the mechanism of action of XYS. Urine obtained from rats subjected to chronic unpredictable mild stress (CUMS) was analyzed by UPLC-MS. Distinct changes in the pattern of metabolites in the rat urine after CUMS production and drug intervention were observed using partial least squares-discriminant analysis. The results of behavioral tests and multivariate analysis showed that CUMS was successfully reproduced, and a moderate-dose XYS produced significant therapeutic effects in the rodent model, equivalent to those of the positive control drugs, venlafaxine HCl and fluoxetine HCl. Metabolites with significant changes induced by CUMS were identified, and 17 biomarker candidates for stress and drug intervention were identified. The therapeutic effect of XYS on depression may involve regulation of the dysfunctions of energy metabolism, amino acid metabolism, and gut microflora changes. Metabonomic methods are valuable tools for measuring efficacy and mechanisms of action in the study of traditional Chinese medicines.

Study of plasma metabolic profiling and biomarkers of chronic unpredictable mild stress rats based on gas chromatography/mass spectrometry.

A metabolomic investigation of chronic unpredictable mild stress (CUMS) rats was carried out. Plasma obtained from Sprague-Dawley (SD) rats treated by CUMS was analyzed using gas chromatography/mass spectrometry. Thirty-seven metabolites were identified among the detected compounds. Subsequent data analysis using the t test and principal component analysis (PCA) revealed significant metabolic changes in the rats' plasma after CUMS treatment. Clear separation between the model and control group was achieved, and the level of twelve metabolites, including amino acids, sugar, organic acids and fatty acids, were significantly different between plasma samples from the controls and CUMS group. These observations suggested that the depressed state may be associated with perturbation of amino acid metabolism, energy metabolism and glycometabolism. The study suggested that the metabolomics approach could be used as a potential powerful tool to investigate the biochemical change in certain physiopathological conditions, such as depression, as an early diagnostic means.

Association of Tongue Bacterial Flora and Subtypes of Liver-Fire Hyperactivity Syndrome in Hypertensive Patients.

Structural changes in symbiotic human microorganisms can affect host phenotype. Liver-fire hyperactivity syndrome (LFHS) presents as bitter taste, halitosis, xerostomia, odontalgia, and other oral symptoms. LFHS is associated with hypertension (EH). In this study, tongue flora was analyzed to further understand the intrinsic relationship between tongue flora and LFHS. Samples of tongue coating, from 16 patients with EH-LFHS, 16 with EH-non-LFHS, and 16 controls, were obtained; then, 16S rRNA variable (V3-V4) regions were amplified and sequenced by MiSeq PE300 Sequencing. Tag clustering and Operational Taxonomic Units (OTUs) abundance analysis were used to compare the OTU sequence with the 16S database. The species were classified, and diversity and structure of the bacterial flora were compared between the three groups. Alpha diversity analysis, including Observed Species index and Chao index, indicated significantly higher richness of species in patients with EH-LFHS (p < 0.05). Higher phylogenetic diversity, in patients with EH-non-LFHS, indicates greater differences in evolutionary history than in patients with EH-LFHS. Streptococcus, Rothia, Neisseria, and Sphingomonas were the most prevalent in patients with EH-LFHS, differed from the other two groups. This indicates that richer bacterial diversity, and structure associated with EH-LFHS, may affect the occurrence, development, and outcome of hypertension and syndrome subtypes recognized by TCM.

Analysis of mutations of two Gitelman syndrome family SLC12A3 genes and proposed treatments using Chinese medicine.

OBJECTIVE: To determine the gene location of two Gitelman syndrome (GS) family SLC12A3 genes and explore treatments using Chinese medicine (CM) prescriptions. METHODS: In order to locate the two GS mutations, samples were collected from 11 people from two different pedigrees for direct genetic sequencing and comparison of the 26 exons of SLC12A3. Furthermore, the change of serum potassium was monitored throughout the therapy and those two probands undertook a sequential superposition of Western medicine (including potassium, Panangin and potassium-sparing diuretics) with CM prescription based on Buyang Huanwu Decoction () and Sijunzi Decoction (). The treatment included three stages, oral potassium chloride for the first 2 weeks (stage 1), potassium-sparing diuretic and Panangin with potassium chloride for the next 2 weeks (stage 2), CM along with the medicine in stage 2 for the final 2 weeks (stage 3). RESULTS: The three mutations occurring in proband 1 from pedigree I were Thr60Met, 965-1_976del13ins12 (small indels mutation) and Ala122Ala (homozygous silent mutation). Likewise, three mutations, Asn359Lys, Thr382Met and Arg913Gln, appeared in the proband 2 from pedigree II. The serum potassium levels increasing from baseline to sequential stages were 1.63 mmol/L (baseline), 2.5 mmol/L (stage 1), 3.1 mmol/L (stage 2) and 3.9 mmol/L (stage 3) in the proband 1, and 2.8 mmol/L (baseline), 3.1 mmol/L (stage 1), 3.5 mmol/L (stage 2) and 4.3 mmol/L (stage 3) in the proband 2, respectively. The symptoms (numbness of limbs, weakness, palpitations, etc.) of both probands were all alleviated. CONCLUSIONS: The mutations of both GS pedigrees can be defined as compound heterozygous mutations, most of which are known as missense mutations. Applying CM could be an appropriate choice for future intervention of GS.

The Construction of Unsmooth Pulse Images in Traditional Chinese Medicine Based on Wave Intensity Technology.

Unsmooth pulse is one of the most important pulses in TCM diagnostics. We constructed the wave intensity (WI) images of unsmooth pulse based on the pressure wave (P), flow velocity wave (U), and WI [(dP/dt)(dU/dt)] by ALOKA Prosound α 10 Color Doppler. The characteristic of Cunkou normal pulse could be summarized as follows: compared to Renying pulse, its W1 amplitude is smaller and NA wave is more obvious, while the W2 wave is indistinct or even invisible, and the R-1st is longer than that of Renying pulse. The principal U wave of Renying pulse looks like "Λ" shape, while it looks like an arched blunt "∩" shape in Cunkou pulse, and the amplitude of U wave in Cunkou pulse is smaller. The direction of the principal U wave in Cunkou unsmooth pulse is up, which shows hoof boots "h" shape with high amplitude and a significant notch on declined branch; the amplitude of predicrotic wave in unsmooth pulse P wave is significantly higher, which could be even higher than that of h1, resulting in early appearance of h3 or integrating with h1, which forms a wide and blunt peak. Unsmooth pulse shows poorer vascular elasticity and greater vascular stiffness.