Objective; To establish a method for content determination of total saponins in Rubus parvifolius L.. Methods; Total saponins was determined by UV - VIS spectrophotometry after different extracts of the sample had been coloured. Results; The method was linear in the range of 22-176?g (r = 0.9997),and the average recovery was 102. 1 % , RSDwas 2. 82% (n = 6). The content of total saponins was highest by the method of 70% ethanol reflux. Conclusion; The sample is stable after coloured. The method is sensitive and reliable , which may be used for the quality control of Rubus parvifolius L. .

Objective:To develop a method for the isolation and purification of suavissimoside R1 from Herb Rubi Parvifolii. Methods:Macroporous adsorption resin and preparative RP-HPLC,the water extract sedimentated by ethanol was pre-isolated by adsorption resin,and then fractionated by column chromatography on silica gel ( 100-200 mesh )using chloroform-methanol-water-acetic acid(40:10:1:0.4%).The collected fraction was purified for suavissimoside R1 by preparative RP-HPLC were combined as pre- pareation method.Results:Suavissimoside R1 was identified by mass spectrometry(MS),infrared spectrometry(IR),ultraviolet spectrometry(UV),nuclear magnetic resonance (NMR) and high performance liquid chromatography(HPLC),and compared with the relevant literature.The final purity of the compound was more than 98%.Conclusion:The developed method is simple,and reproducible.The solvent is cheap,low in boiling point and easily recovered.The product of suavissimoside R1 can be used as reference substance for analysis.

Objective To investigate the effects of Liuwei Hezhong Powder on gastrointestinal motility and hormones,and to explore its possible mechanism.Methods The small intestinal propulsion ratio,endogastric residual rate and total gastrointestinal propulsion time in mice were observed after intragastric administration of Liuwei Hezhong Powder .Radioimmunoassay was applied to detect the changes of gastrin,motilin and vasoactive intestinal polypeptide in spleen asthenia rats.Results Compared with the blank control group,Liuwei Hezhong Powder can significantly improve the small intestinal propulsion,reduce endogastric residue and shorten total gastrointestinal propulsion time(P 0.05),but the high dosage group of Liuwei Hezhong Powder were especially remarkable(P

Aim To explore the mechanism of neuro-protecting effects of TGRP on ischemic cerebral injury.Methods The middle cerebral artery was occluded for 2 h to produce focal ischemic followed by 24 h reperfusion.HE staining,TUNEL labeling and immunohistochemical methods were used to investigate changes of neuronal apoptosis and expression of the apoptosis-related proteins after administration of TGRP.Result TGRP decreased the pathologic changes significantly.The apoptosis of neural cells and the expression of Bax positive cells in the TGRP groups significantly decreased compared with the control group(P

Objective:To explore the expression and location of TLR5 and NLRC4 on different breast cancer cell lines MDA-MB-231,MCF-7 and MDA-MB-435 and TLR5 activation in breast cancer cell line by recombinant flagellin . Methods:The mRNA level of TLR5 and NLRC4 in MDA-MB-231, MCF-7 and MDA-MB-435 cell were detected with quantitative Real-time PCR and TLR5 expression and location in MDA-MB-231 and MCF-7 cell were detected with Flow cytometry. Induction,expression,purification and i-dentification of recombiant flagellin,including FliC (activating both TLR5 and NLRC4),FliC△90-97(unable to activate TLR5),FliC-L3A (unable to activate NLRC4),FliC△90-97:L3A (unable to activate both TLR5 and NLRC4). 1 μg/ml recombinant flagellin were used to stimulate MCF-7 cell lines,12 h later,the supernate were collected,and ELISA was performed to assess the secretion of IL-8. Results:The mRNA level of TLR5 in MCF-7 cell was 1 700 folds higher than that of MDA-MB-435. TLR5 was expressed in MCF-7 cell surface and ctyosol,while expressed only in cytosol in MDA-MB-231 cell. FliC and FliC-L3A,which were able to activate TLR5 pathway,stimualted MCF-7 cell line to secret IL-8,but FliC△90-97 and FliC△90-97:L3A did not. Conclusion:TLR5 and NLRC4 have been expressed in different breast cancer lines,but there exists difference on the expression level and location of TLR5. Expression level of TLR5 and NLRC4 in MCF-7 cell were higher than other breast cancer lines. TLR5 receptor which is expressed on the surface of breast cancer cell can be activated by flagellin,and these work also provide us experimental basis to further understand the impact of TLR5 activation on breast cancer cell proliferation.

Objective To extract,isolate and identify the polysaccharide from ginseng,and to investigate its anti-tumor activity in vitro. Methods The ginseng polysaccharide was obtained through water extraction and ethyl alcohol deposition method. Use the Sevage method to remove the protein in the crude polysaccharide. The structural characteristics of the polysaccharide were determined by FT-IR spectra.The RM-1 and HeLa cells were divided into control group and different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )of ginseng polysaccharide groups. The survival rates of the cells in various groups were detected by MTT method. The indirect killing effects of ginseng polysaccharide with different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )on the cancer cells were determined by CTL test. Results The extraction rate of ginseng polysaccharide was 8.7 6% and the structural characteristics demonstrated that the main component of the extract was polysaccharide.The result of MTT showed that there were no significant differences of the survival rates of RM-1 and HeLa cells between different concentrations of ginseng polysaccharide groups after treated for 24 h compared with control group (P>0.05).The result of CTL test showed that the cytototic LHD release rates in different concentrations of ginseng polysaccharide groups were increased significantly (P<0.05)compared with control group;with the increase of ginseng polysaccharide concentration, the cytotoxic LDH release rates were increased firstly and then were decreased.When the concentration of ginseng polysaccharide was 100 mg·L-1 ,the cytotoxic LDH release rate was the biggest.Conclusion Ginseng polysaccharide can indirectly inhibit the growth of the tumor cells by activating T cells and play an anti-tumor effect.

Objective To extract the Ginseng polysaccharide (GPS), polysaccharides of Tricholoma matsutake (PTM)and polysaccharide of Lentinus edodes (PLE)from gingeng, tricholoma matsutake and lentinus edodes respectively,and to analyze and identify their structures,and to prepare their complex,and to study the indirect antitumor activity invitro of polysaccharide complex.Methods The polysaccharides were extracted with hot water and precipitated by ethanol.The carbohydrate levels were determined by the method of phenol-sulfuric acid.The m-hydroxyphenyl method was used to determine the levels of uronic acid, and the national standard method was used to determine the levels of starch.Infrared spectroscope and chemical methods were performed to analyze their structures. Orthogonal experiment was used to study mixing methods. Cytotoxic T lymphocyte experiment and LDH release assay were performed to detect the influence of polysaccharide complex of GPS,PTM,and PLE in the CTL killing activity,and its indirect killing effect on the P815 cells.Results The extraction rates of GPS,PTM, and PLE were 8.85%,9.40%,and 10.50%;the levels of total polysaccharides were 62.96%,59.13%,and 33.86%;the levels of uronic acid were 16.44%,9.37%,and 16.44%;the starch levels were 7.26%,2.80%,and 3.77%,respectively.The identification results showed that the polysaccharides were obstrained.When the quality ratio of the three kinds of polysaccharides was 1∶1∶1 and the concentration was 600 mg·L-1 ,the CTL cytotoxicity was the highest.Conclusion The polysaccharide complex is obtained,identified and characterized. Polysaccharide complex can enhance the cytotoxicity of CTL and has the indirectly inhibitory effect on the proliferation of P815 cells.

Objective:To investigate the role of C3a,C5a and their receptors in the pathogenesis of IgA nephropathy (IgAN). Methods:A total of twenty-eight 6-8 weeks old female BALB/c mice were investigated.And they were negative control group , WT group,C3aR-/-group,C5aR-/-group(the latter three groups were named as experimental groups ),seven mice in each group.All the mice were infected through respiratory tract with infectious SV (experimental groups) or PBS(negative control group),combined with tail vein challenge to make IgAN animal model.Testing 24 h total urinary protein , serum urea nitrogen ( BUN ) and creatinine ( Cr ) , using direct immunofluorescence to test the renal deposition of IgA and C 3,observing renal pathologic lesion under PAS staining with light microscopy.RT-qPCR was used to test the relative mRNA expression of TNF-α,TGF-β,IL-1β,IL-6,MCP-1.Results: After 15 weeks,the level of UTP in experimental group was significantly higher than negative control group ,and the same results as WT group than C3aR-/-group and C5aR-/-group.There was no significant difference among groups for BUN and Cr.Combined with negative control group , experimental groups had significant renal pathological lesions , and the changes of WT group was more severe than C3aR-/-group and C5aR-/-group.The results of relative mRNA expression of TNF-α,TGF-β,IL-1β,IL-6,MCP-1 was the same as the level of 24 h UTP,at the same time,the relative mRNA expression of IL-1β,IL-6,MCP-1 in C3aR-/-group was significantly less than C5aR-/-group.Conclusion:The deficiency of C3a/C5a receptors can protect kidney from injury in IgAN ,and C3a receptor has more significant role in protect kidney from injury in IgAN.

Objective To analyze karyotype,genetic characteristics and clinical features of Turner syn-drome.Methods Cytogenetic analysis and genetic counseling were performed for patients with the diagnosis of Turner syndrome.Results Analyzed karyotype of forty-nine patients with Turner syndrome,there are five major categories such as simple type,numerical abnormality and chimeras,structural abnormality and chime-ras,both numerical abnormality and structural abnormality chimeras,contained the Y chromosome.Abnormal karyotypes were presented at eighteen species.Simple type was the most prevalent type of patients with Turn-er syndrome(43%).Conclusion Different karyotype exhibit some different phenotype.The clinical manifesta-tion of chimeras might depend on the proportion of abnormal karyotype,Case contained the Y chromosome would be hermaphrodismy mostly.Timely diagnosis of the disease would have positive significance to preven-tion and therapy.Hormonal therapy could improve patient's height and gonad development,the case contained the Y chromosome should pay attention to prevent gonadoblastoma.

OBJECTIVEThis study aimed at determining the influence of hydrofluoric acid (HF) in varied concentrations on the surface morphology of lithium disilicate glass ceramics and bond durability between resin composites and post-treated lithium disilicate glass ceramics.

METHODSAfter being sintered, ground, and washed, 72 as-prepared specimens of lithium disilicate glass ceramics with dimensions of 11 mm×13 mm×2 mm were randomly divided into three groups. Each group was treated with acid solution [32% phosphoric acid (PA) or 4% or 9.5% HF] for 20 s. Then, four acidified specimens from each group were randomly selected. One of the specimens was used to observe the surface morphology using scanning electron microscopy, and the others were used to observe the surface roughness using a surface roughness meter (including Ra, Rz, and Rmax). After treatment with different acid solutions in each group, 20 samples were further treated with silane coupling agent/resin adhesive/resin cement (Monobond S/Multilink Primer A&B/Multilink N), followed by bonding to a composite resin column (Filtek™ Z350) with a diameter of 3 mm. A total of 20 specimens in each group were randomly divided into two subgroups, which were used for measuring the microshear bond strength, with one of them subjected to cool-thermal cycle for 20 000 times.

RESULTSThe surface roughness (Ra, Rz, and Rmax) of lithium disilicate glass ceramics treated with 4% or 9.5% HF was significantly higher than that of the ceramic treated with PA (P<0.05). The lithium disilicate glass ceramics treated with 9.5% HF also demonstrated better surface roughness (Rz and Rmax) than that of the ceramics treated with 4% HF. Cool-thermal cycle treatment reduced the bond strength of lithium disilicate glass ceramics in all groups (P<0.05). After cool-thermal cycle, the lithium disilicate glass ceramics treated with HF had higher bond strength than that of the ceramics treated with PA. The lithium disilicate glass ceramics treated with 4% HF had higher bond strength than that of the ceramics treated with 9.5% HF (P<0.05). During cool-thermal cycle, the lithium disilicate glass ceramics treated with 4% HF demonstrated higher reduction in bond strength than that of the samples treated with 9.5% HF (P<0.05).

CONCLUSIONSThe concentration of HF significantly affected the surface morphology of lithium disilicate glass ceramics and the bond durability between resin composites and post-treated lithium disilicate glass ceramics. The bond strength between resin composites and post-treated lithium disilicate glass ceramic was more efficiently maintained by treatment with 9.5% HF.