Study on Difference of Intestinal Flora in Patients with Primary Insomnia with Different TCM Syndromes / 中国中医药信息杂志

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.

Analysis of mutations of two Gitelman syndrome family SLC12A3 genes and proposed treatments using Chinese medicine / 中国结合医学杂志

<p><b>OBJECTIVE</b>To determine the gene location of two Gitelman syndrome (GS) family SLC12A3 genes and explore treatments using Chinese medicine (CM) prescriptions.</p><p><b>METHODS</b>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).</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Methylation in Promoter Region of SLC6A2 Gene in Heart Failure Patients and Its Correlation with Qi Deficiency/Blood Stasis Syndrome / 中国中西医结合杂志

<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>

Design, synthesis and activities of novel benzothiazole derivatives containing arylpiperazine / 药学学报

Twenty-four novel benzothiazole derivatives containing arylpiperazine were designed and synthesized by bioisosterism principle. Anti-proliferative effect of these synthesized compounds against four cancer cell and two normal cell lines were evaluated in vitro by the standard MTT assay. Pharmacological test showed that most of the compounds exhibited potent antitumor activity. Some of the compounds (II2, II3, II6, II7) showed strong anti-proliferation activities against HepG2 and HeLa229 cell lines with the IC 50 values of 1.6-4.5 micromol x L(-1) and 2.5-5.3 micromol x L(-1), respectively, and compounds having cyan in p-substituted benzene ring (I4, I8, I12, II4, II8 and II12) were found to have better antitumor activities against AsPC-1 cell lines with the IC50 values of 5.2-11.3 micromol x L(-1). The structure-activity relationship of benzothiazole derivatives containing arylpiperazine was also discussed preliminarily.

Relationship between gene polymorphism and haplotype of SLC6A2 promoter and blood-stasis pattern in patients with essential hypertension / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the relationship between the genetic variants of the norepinephrine transporter gene solute carrier family 6 member 2 (SLC6A2) and blood stasis pattern (BSP) in patients with essential hypertension (EH).</p><p><b>METHODS</b>DNA was extracted from the peripheral blood of 830 EH patients (532 were typed as BSP and 298 as non-BSS) and 512 persons with normal blood pressure (for control), to detect the polymorphisms of SLC6A2 promoter-3 and 2 by PCR-RFLP, and estimate the haplotype frequency adopting SHEsis program.</p><p><b>RESULTS</b>Chi2 partition showed that frequency of promoter-2-GG genotype in EH patients of BSP was lower than that in the EH patients of non-BSP and the normal control (P = 0.001); while that of promoter-3-GG/GA in the EH patients with severe BSP was the highest (P < 0.001). Logistic regression analysis showed that after the miscellaneous factors being rectified, with the reference category of EH-non-BSP, the relative odds ratio (OR) of promoter-2-GC/CC genotype for EH-BSP was 1.535 (95% CI: 1.094-2.155, P = 0.013); that of promoter-3-AG/GG genotype for EH with severe BSP was 1.925 (95% CI: 1.199-3.091, P = 0.007); while OR of G-C haplotype (promoter-3-promoter-2) for EH-BSP was 2.127 (95% CI: 1.202-3.765, P = 0.010), showing the strongest intensity.</p><p><b>CONCLUSION</b>SLC6A2 promoter-3-GA/GG genotype may be a susceptibility gene for patients of EH with severe BSP; promoter-2-GC/CC and G-C haplotype might be the susceptible factors to EH-BSP.</p>

Mutations of ACVRL1 gene in a pedigree with hereditary hemorrhagic telangiectasia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify the activin A receptor type II-like 1 gene (ACVRL1) mutations in a Chinese family with hereditary hemorrhagic telangiectasia (HHT2).</p><p><b>METHODS</b>The exons 3, 7 and 8 of ACVRL1 gene of the proband and her five family members were amplified by polymerase chain reaction (PCR), and the PCR products were sequenced.</p><p><b>RESULTS</b>The proband had obvious telangiectasis of gastric mucosa, and small arteriovenous fistula in the right kidney. All the patients in the HHT2 family had iterative epistaxis or bleeding in other sites, and had telangiectasis of nasal mucosa, tunica mucosa oris and finger tips. ACVRL1 gene analysis confirmed that there is frameshift mutation caused by deletion of G145 in exon 3 in the 4 patients, but the mutation is absent in 2 members without HHT2.</p><p><b>CONCLUSION</b>The HHT2 family is caused by a 145delG mutation of ACVRL1 gene, resulting in frameshift and a new stop codon at codon 53.</p>

Clinical analysis on high-sensitivity C-reactive protein in hypertensive patients with blood-stasis syndrome / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the correlation of serum high-sensitivity protein (hs-CRP) level and the clinical score of blood-stasis syndrome as well as the clinical indexes in hypertensive patients.</p><p><b>METHODS</b>Levels of serum hs-CRP, blood lipids, fasting blood glucose (FBG), body mass index (BMI), left ventricular mass index (LVMI), fibrinogen (Fg), urinary albumin (UA), and score of blood-stasis syndrome (SBSS) in 117 patients were measured, and the relationship between hs-CRP and the other indexes was analyzed.</p><p><b>RESULTS</b>Spearman rank correlation analysis showed that hs-CRP level was significantly positively correlated with SBSS, triglyceride (TG), FPG, BMI, LVMI, Fg, and UA, while negatively correlated with high density lipoprotein cholesterol (HDL-C).</p><p><b>CONCLUSION</b>Hs-CRP is closely related with the formation and development of blood-stasis syndrome in hypertensive patients.</p>