A stragalus membranaceus(AM ) plays an important role in the traditional Chinese recipe" Decoction of Invigorating Yang for Recuperation" (DIC ) as the main ingredient which invigorates Qi and promotes blood circulation. But in clinics,doses of AM used in tbe recipe of ten varies. For the purpose to investigate the effect of different doses of AM on the action of DIC, animal experiments were designed and studied. Rsults showed that when the doses of AM were 15 and 30g,the resulting DIC showed a better antiirritability,and enhancing immunity effects,as well as in vivo antithrombosis and vitro anticoagulation actions as compared to a dose of 120g AM in DIC (P

Objective To optimize the conditions of extraction process of polysaccharides from seeds of Toona sinensis; To preliminary determinate the antioxidant activity in vitro. Methods The content of polysaccharides from Toona sinensis was chosen as evaluation index; the extraction time, material liquid ratio, and extraction times were chosen as factors; L9(34) orthogonal design was used to optimize extraction process; antioxidant activity of polysaccharides from Toona sinensis was investigated by the total reducing power and 1,1-diphenyl-2-picryhydrazyl (DPPH) method. Results From the statistical data of processing, the optimal extracting conditions were as follows:liquid to material was 1:12, extraction time was 120 min, and extracted for 3 times. Polysaccharides from seeds of Toona sinensis had certain reducing capacity and had better scavenging ability on DPPH? and the scavenging rate significantly increased with the concentration. The effect was simulated by curve equation. Conclusion The process is simple, feasible, stable and reliable and can be used for the small-scale study on extraction of polysaccharides from seeds of Toona sinensis. Polysaccharides from seeds of Toona sinensis have antioxidant activity.

Objective To study the effects of high albumin, high glucose and low bovine serum on the phenotypic transformation of renal tubular epithelial cells. Methods The normal human kidney proximal tubular eel] line (HKC) was cultured for 30 days in the presence of high albumin (1.5 g/L), high glucose(25 mmol/L) and low bovine serum(2% ) . Morphological changes were observed by electronic microscopy. Immunohistochemistry stain was used to examine the expression of cytokeratin, vimentin, a-SMA, collagen Ⅰ and TGF-pl protein. Western blot was applied to further detect the process of collagen I protein expression, and in situ hybridization was used to examined the expression of collagen Ⅰ gene. Results Renal tubular epithelial cells cultured in high albumin, high glucose and low bovine serum showed obvious morphologic changes, including elongated shape, decrease of microvilli and mitochondria, and increase of rough endoplasmic reticulum under electronic microscopy. Immunohistochemistry stain revealed the reduction of cytokeratin, and enhancement of vimentin, ?-SMA, TGF-?1 and collagen Ⅰ. Western blot demonstrated that the expression of collagen Ⅰincreased in a time-dependent manner, and in situ hybridization showed that collagen type Ⅰ mRNA increased as well. Conclusion High albumin, high glucose and low bovine serum induce phenotypic transformation of renal tubular epithelial cells into mesanchymal cells.

Objective: Cellular proliferate inhibition, senescence or apoptosis are induced by telomere shortening through the activation of P53 pathway, but so far, little is known of the mechanism. This study aimed to clarify the molecular regulation of P53 through telomere pathway by the investigation of molecular interaction between P53 and the main telomere associated protein telomeric repeat binding protein 1(TRBP1) in vitro. Methods: Glutathione S-transferase (GST) alone and 4 different human P53-GST fusion proteins were expressed in E. coli. and purified through glutathione Sepharose TM 4B by affinity chromatography, P53s were wild type P53 (1-393), N terminal truncated form P53 2C (95-393), C terminal truncated form P53 N5 (2-293) and single amino acid mutant P53 R175H (175 arginine to histidine). Glutathione Sepharose TM 4B, purified GST alone and P53 fusions were mixed with human breast cancer cell line MCF-7 cellular protein extracts through in vitro binding assay-pull down, the molecular interaction between P53 and TRBP1 were detected by Western blot. Results: SDS-PAGE and Coomassie brilliant blue staining showed that the molecular weights of all the purified proteins were as expected and purities were over 90%. Western blot of TRBP1 showed that both wild type P53 and P53 R175H could bind to TRBP1 of MCF-7 cells, and their binding capacities are similar, whereas GST alone and Glutathione Sepharose TM 4B beads couldn’t. Compared with both of them, the interaction between P53 2C and TRBP1 enhanced dramatically, but between P53 N5 and TRBP1 reduced significantly.Conclusion: P53 can interact with TRBP1 directly and in vitro, C terminus of P53 (293-393) is the structural domain of their interaction. This C terminus domain dependent interaction between P53 and TRBP1 may be related to the cellular activities induced by telomere dynamic changing.

Objective To preliminary establish the process condition of extracting polysaccharides from Allii tuberosi semen.Methods The study was carried out with the content of Semen allii tuberosi polysaccharides as evaluation index, the material liquid ratio, extraction time, and extraction times as observation factors, and L9 (34) orthogonal design as extraction craft.The physical and chemical properties and IR spectroscope were used to identify the extract.Results The optimum preparation condition was determined as: using 12 times water, extracting 3 times, and extracting 160 minutes.According to the results of purple ring reaction and infrared spectrum, the extract from Allii tuberosi semen has polysaccharides.From the statistical processing of data we found when polysaccharides from Semen allii tuberosi has 50％ reducing power, the value of EC50 is 10.2 mg/ml.Conclusion The process is reasonable and suitable and can be used for the extraction of polysaccharides from Allii tuberosi semen, and can provide the theoretical basis for the research and application.

Objective To detect the effect of oridonin(ORI)on the gene expression of human esophageal carcinoma cell SHEEC and to screen the tumor cell apoptosis target genes.Methods The gene expression of human esophageal carcinoma cell SHEEC without and with ORI induction for 1 hours and 8 hours were detected with microarray technique,respectively.The differentially expressed genes were identified and verified with fluorecent quantitative PCR.Results A total of 1011 genes showed up or down regulation more than twice after ORI induction(including 280 genes after 1 hour and 731 genes after 8 hours induction respectively).In these genes,17 genes with the top extent of up or down regulation were identified,which were involved in the cell signal transduction,transcription regulation,and cell apoptosis.These 17 differentially expressed genes were verified with real-time PCR,and 12 genes were statistically significant.Conclusion In the 12 differentially expressed genes with statistically significance,there may have tumor cell apoptosis target genes induced by ORI through mitochondrion route.

AIM: To study the absorption performance and purification process of total flavones from Bushen Recipe(Radix polygoni multiflori;Herba epimedii;Rhizoma drynariae;etc) with macroporous resin. METHODS: The contents of total flavones,icariin,tetrahydroxystilbene and naringin were worked as indexes,the adsorptive capacity of D101,DA201 and DM301 resins were investigated,and the static adsorption curve,dynamic leakage curve and elution curve were used for the selection of macroporous resin. RESULTS: The elution parameters of selected DM301 consisted of the 0.25 BV/h feed rate solution ratio of 4 BV water to elute resin and 4 BV 70% alcohol to be collected as elution solvent,and the elution rate of 2.4 BV/h was established. CONCLUSION: Macroporous resin method is a better candidate method for purifying flavones in Bushen Recipe.

Objective To analyze the relationship between genetic variability and evolution among Trypanosoma brucei (including T. b. brucei, T. b. rhodesiense and T. b. gambiense), T. evansi and T. equiperdum isolates. Methods Genomic DNAs of 26 trypanosome isolates were amplified by a mobile genetic elements (MGE)-PCR technique and cluster analysis was performed based on the molecular profiles with Neighbor-Joining method. Results The genetic variability among trypanosome isolates examined was obvious with an average genetic distance of 41.2% (ranged from 0 to 100%). Similarity coefficient among T. brucei isolates was 41.15% which was lower than that between T. evansi and T. equiperdum isolates. The closest relationship was found between T. evansi and T. brucei isolates with a similarity coefficient of 62.94%. The genetic variability between T. b. rhodesiense and T. b. brucei isolates was higher than that among T. b. gambiense isolates. Conclusion Species and subspecies in Trypanozoon displayed a higher genetic variability; T. equiperdum isolates collected from China and from South America, and T. evansi isolates from China and from South America, should have a similar origin.

AIM: To study on the differences of chemical compos i tion between extracts of Bushen recipe prepared by decocting together and single , with fingerprint technology. METHODS: The extracts of Bushen recipe were prepared by decocting together and single under the same process. The comparison was done under the different wavelength s and different brand columns, HPLC method determined the extract's fingerprint within the range of optimized conditions. RESULTS: The ratio of The compositions and its main chemical substa nces are the different between the two extracts, and the extract rate of the main chemical substances are higher in extrat decocted single than in extract decocted togethe r. CONCLUSION: There are some chemical substances and composition's ra tio differences in the extracts of Bushen recipe made by decocting together and single.