Genomewide DNA Methylation Responses in Patients with ß-Thalassemia Treated with Yisui Shengxue Granules ().

OBJECTIVE: To examine the clinical effects of Yisui Shengxue Granules () in the treatment of ß-thalassemia and explore its mechanism on DNA methylation levels. METHODS: A randomized placebo-controlled double-blinded trial was conducted. Forty patients with ß-thalassemia were recruited and distributed randomly by envelope method into an experimental group and a control group, 20 patients in each group. The patients were given Yisui Shengxue Granules in the experimental group and placebo in the control group (12 g/bag three times a day) during a 3-month intervention. Before and after 1, 2, and 3 months of treatment, peripheral intravenous blood was sampled, and blood parameters such as hemoglobin (Hb), red blood cells (RBCs), reticulocytes (Ret), and fetal hemoglobin (HbF) were analyzed. Mononuclear cells from 5 patients, who showed an obvious treatment effect, were isolated by density gradient centrifugation. DNA methylation was analyzed using an Affymetrix USA GeneChip Human Promoter 1.0 Array and Input-promoter 1.0. RESULTS: Compared with pre-treatment, there was an obvious increase in Hb and RBCs counts after 1, 2, and 3 months in the experiment group (P<0.01 or P<0.05). Meanwhile, HbF increased from the 2nd to the 3rd month (P<0.05). In the control group, Hb and RBCs showed no obvioas change. After 3-month treatment, DNA methylation results from 5 patients revealed that there were 24 hypomethylated genes and 3,685 hypermethylated genes compared with pre-treatment. Genes of insulin-like growth factor 1 receptor (IGF1R) and Janus kinase 3 (JAK3) revealed the most relations with other genes (degree: 21) and genes of 1-phosphatidylinositol-4, 5-bisphosphate phosphodiesterase gamma 2 (PLCG2) and mitogen-activated protein kinase 10 (MAPK10) showed a stronger intermediary role (betweenness centrality=0.04). CONCLUSIONS: JAK3 and MAPK10 are two key genes in bone marrow and the lymphatic system, and JAK3 is likely to be related to hematopoietic cytokines in the process of early hematopoiesis. (Registration No. NCT01549080).

Clinical Effect and Mechanism of Yisui Shengxue Granules in Thalassemia Patients with Mild, Moderate, or Severe Anemia.

Yisui Shengxue granules, which is a Chinese traditional medicine, can increase hemoglobin, red blood cells, and Ret of thalassemia patients with mild, moderate, and severe anemia and thus relieve clinical anemia symptoms. Studies on mechanism found that Yisui Shengxue granules can increase the proliferation ability of hematopoietic stem cells. Emodin promoted colony forming of hematopoietic stem cells. Yisui Shengxue granules can increase the activity of GSH-PX in bone marrow blood and decreased the severity of inclusion bodies on the cytomembrane of RBCs. YSSXG attenuated anemia symptoms in patients with thalassemia mostly by increasing the proliferation of hematopoietic stem cells and decreasing the hemolysis of RBCs.

[Comparative Study on Boiling Powder and Traditional Herbal Pieces of Rhubarb].

Objective: To optimize the particle size of rhubarb boiling powder, to determine decoction process of rhubarb boiling powder through the comparative study with traditional herbal pieces. Methods: Three kinds of particle size of rhubarb boiling powder were prepared. The content of aloe emodin,rhein, emodin, chrysophanol and emodin monomethyl ether were detected by HPLC,and to determine particle size of rhubarb boiling powder according to the content of total chemical ingredients and dry extract rate in water decoction. The effect of the amount of water and decocting time on decoction of boiling powder and traditional herbal pieces of rhubarb were systematically investigated by using single factor test. Results: Firstly, the content of 2 mm to 0. 18 mm particles in rhubarb boiling powder can not be less than 80%. Total anthraquinone content and dry extract rate had no significant difference in the decoction of boiling powder in the amount of water between 14 ~ 20 times; decocting time had no significant effect on total anthraquinone and dry extract rate; the content of combined anthraquinone decreased significantly, while the decocting time was more than 20 min. The decocting time had great influence on the herbal pieces, with ten times of water,and decocting 5 ~ 50 min each time, the compound content and dry extract rate of boiling powder were 3. 23 ~ 9. 48 times and 1. 61 ~ 3. 44 times than that of traditional herbal pieces, and with the extension of the decocting time, the gap was narrowing. Conclusion: Preparation process of rhubarb boiling powder is simple and suitable for industrial production, boiling powder can save more than half of the amount of traditional herbal pieces with ten times of water, decocting one times, decoction time of 10 ~ 15 min.

[Comparative Study on Efficiency of Extraction of Boiling Powder and Traditional Herbal Piecesâ . Part of Root and Rhizome,Flower,Leaf and Whole Plant Medicinal Materials].

Objective: To compare the efficiency of extraction of boiling powder and traditional herbal pieces of root and rhizome, flower, leaf and whole plant medicinal materials for providing a reference for the study and clinical application of boiling powder. Methods: 19 kinds of root and rhizome herbs, the 4 kinds of flower medicinal materials, the 2 kinds of leaf medicinal materials and the 2 kinds of whole plant medicinal materials were chosen as the researh object and were crushed into boiling powder. The contents of the active ingredients and dry extract rate in the water decoction were set as the index to compare the extraction efficiency of boiling powder and traditional herbal pieces. Results: Boiling powder was added ten times of water, decocted one time, decoction time was only ten min, the dry extract rate and the concentration of ingredients in the decoction of boiling powder were not less than those in the traditional decoction of herbal pieces( ten times of water, decoction time was 50 min),this difference was mainly caused by the different of texture density and size of the medicinal materials. Conclusion: The boiling powder can not only save time and facilitate the patient, but also save a lot of medicinal materials from the perspective of chemical medicine, which is beneficial to the sustainable development of medicinal material resources.

[Study on preparation process and quality control of jiangtangyin teabag].

OBJECTIVE: To study preparation technology and establish the quality standard of Jiangtangyin teabag. METHODS: An orthogonal test was adopted in this study. The transter rate of puerarin and the dried extraction rate were used as indices for optimizing the extraction condition. Quality standards of Jiangtangyin teabag were established by TLC and HPLC method. RESULTS: The best extraction technology was as follows: The drugs were extracted for 3 times, the ratio of water to solid was 10, 8 and 8, respectirely, the extracting time was 1.5 h, 1 h and 1 h respectively. The speckles of TLC method were distinct, accurate and sensitive. The calibration curve of puerarin was linear in the range of 0.168 - 0.84 microg, r2 = 0.9997. CONCLUSION: The preparation process is reasonable and feasible. The methods of quality control are scientific and can be used to control the quality of the Teabag.