Objective To study the lab-scale simulated method of three-effect dynamic countercurrent extraction from Hedyotis diffusa by multi-index optimization and SPSS software.Methods Polyphenol content,flavonoids content,scavenging activity of samples on DPPH radicals,and extracting rate were considered as the optimization indexes.Orthogonal design method was used to optimize the four main factors of alcohol concentration,ratio of solid to solution,extracting temperature,and extracting times.Results The optimum condition is 70 min extraction time,solid to solution ratio of 1∶14,50% alcohol,extracting temperature 85 ℃.Conclusion The condition of large-scale industrialized production might be explored by studying the lab-scale simulated method of dynamic countercurrent extraction from H.diffusa.

Objective:To study the antioxidant activity and anticancer activity of tea polysaccharide.Methods:Tea polysaccharide was extracted and purified by gel permeation chromatography from coarse old green tea leaves obtained from Wuyuan county of Jiangxi province in China.The antioxidant activity of the tea polysaccharide was evaluated by determining the change in absorbancy after heat-induced oxidation in a linoleic acid system with ?-carotene,the decoloration of the 1,1-diphenyl-2-picrylhydrazyl and the elimination of superoxide generated from autoxidation of pyrogallol.On the other hand,the anticancer activities of tea polysaccharide at different concentrations were evaluated by MTT assay using two colon cancer cell lines(HCT-15,Caco-2).Results:Tea polysaccharide possessed distinctive antioxidative activity.The anti-proliferation capabilities of the tea polysaccharide against the tested human colon cancer cell lines were different under the designed experiment conditions.The tea polysaccharide with high purity exhibited good anti-proliferation activity against HCT-15 at concentration of 7.0?mol/L.Conclusion : Tea polysaccharide has remarkable antioxidative and anticancer bioactivities..

Objective: To study the effect of sulforaphane (SUL) on cell growth inhibition, cell cycle, apoptosis and its mechanism in different breast cancer cell lines. Methods: By means of MTT assay, flow cytometry and Western blotting, the effects of the SUL different concentrations on cell growth inhibition, cell cycle arrest, F3Ⅱapoptosis and expression of p34cdc2 and Cdc25C were studied. Results: (1) SUL had strong inhibition effects on the cell growth of tested mammary cancer cell lines, in which the sensitivity of ERP cell lines to SUL was stronger than that of ERN cell line. (2) SUL exhibited obviously G2/M cell cycle arrest to F3Ⅱ and two kinds of ERP cell lines at 5-10?mol/L, whereas no effect on the cell cycle of ERN cell line. (3)The mechanism of hindering transition of F3 Ⅱ cells from G2 phase to M phase was enhancing the phosphorylation of Cdc2, down-regulating the expression of Cdc25C, and inducing inhibition to thedephosphorylation of cyclin B1-Cdc2 complex. (4) No apoptosis was observed under tested conditions. Conclusion: Sulforaphane exhibited obvious differences in the inhibiting effects on the cell proliferation and cell cycle arrests of four different tested breast cancer cell lines, and did not induce apoptosis in F3Ⅱcell line under tested conditions. The mechanism of cell cycle arrest of F3Ⅱcell line appears to involve enhancing the phosphorylation of Cdc2, and down-regulating the expression of Cdc25C.

The effects of adsorption time, solid-liquid ratio, sample concentration, flow rate on the adsorption properties of supported ionic liquids(the N-methylimidazolium functionalized silica, SilprMin) for flavonoids were investigated. The results indicated that the adsorption equilibrium for tested compounds, genistein, luteolin and quercetin, was achieved within 30 min, the adsorption efficiencies of these three compounds were improved with the increase of solid-liquid ratio, and decreased with the increase of their concentrations. Moreover, the adsorption isotherm data of the three tested compounds were in good concordance with the Langmuir model. The saturated adsorption capacity of SilprMim for genistein, luteolin and quercetin was 47.7, 52.5 and 63.2 mg/g, respectively. Adsorption efficiency of SilprMim could reach more than 90% at a sample flow rate of 0.5～1.5 mL/min. Using methanol as eluent, the saturated desorption efficiencies of genistein, luteolin and quercetin were 86.1%, 83.3% and 84.6%, respectively. The elution order of the three tested flavonoids was genistein, luteolin and quercetin. SilprMim had strong adsorption and separation capacity for three tested flavonoids, which was hopeful to be applied in separation and purification of naturally occurring flavonoids.

Objective To explore effect of quercetin on growth of cervical cancer,and its relationship to expression heparanase, and to investigate the possible mechanism of the inhibition effect of quercetin on cervical cancer .Methods HeLa cells were injected into nude mouse to establish the model of cervical cancer.30 nude mice with palpable tumors were randomly assigned to five groups:25,50,100 mg/kg quercetin groups,PI-88 positive group and blank control group.Transmission electron microscope was used to observe the ultrastructural changes of the vascular endothelial cells in each group.The expression of heparanase protein was detected by Western blot.Used blank control group for grey reference, relative grey value of each group was calculated for the further statistical analysis.Results 50,100 mg/kg quercetin groups and group of PI -88 made the volume of tumor tissues smaller.In 50 mg/kg quercetin group, swelling of VEC and mitochondria in tumor tissues was observed.In 100 mg/kg quercetin group, the disappearance of membraneaceous structures was found.In PI-88 positive group, the atresia of lumen, vacuole-shaped smooth endoplasmic reticulum and a dead cell were observed.50 and 100 mg/kg quercetin groups decreased the heparanase protein levels significantly(P<0.05).Furthermore, PI-88 positive group could decrease the heparanase protein levels significantly(P<0.05), compared to 50 and 100 mg/kg quercetin groups.However, the difference between the 25 mg/kg quercetin group and blank control group showed no statistical significance.Conclusion The growth of cervical cancer could be inhibited by 50, 100 mg/kg quercetin.Meanwhile, the ultrastructural changes of tumor endothelial cells could be induced by 50,100 mg/kg quercetin.These changes may relate with the decrease of heparanase protein level .

Objective: The effects of purple-fleshed sweet potato anthocyanin (PSA) on anti-oxidation and anti-aging were investigated. Methods: The effects of different doses of PSA(100、 500、1000 mg/kg bw)on the serum T-AOC were investigated in aged mice after administration for 3 d, 10 d and 18 d. The content of MDA, the activity of serum SOD and blood GSH-Px were measured, and compared with those in Vit E and control group after 30 d. Result: PSA significantly increased the serum T-AOC in aged mice, and this increment was higher when given longer. PSA significantly decreased MDA and enhanced SOD and GSH-Px activities. The efficiency on antioxidative capacity of aged mice at PSA 100 mg/kg bw was equivalent to that of vitamin E at the same amount (100 mg/kg bw). The antioxidation state of 13 mo mice given PSA at 1000 mg/kg bw was near to that of 4 mo mice. Conclusion: PSA has marked effect of antioxidation and can delay aging.

Objective To explore the function and clinical effect of automatic nerve retractor in micro-endoscopic discectomy.Methods From August 2009 to December 2010,350 patients with lumbar disc hemiation were treated by micro-endoscopic discectomy,including 196 males and 154 females,aged from 17 to 68 years (average,42 years).Three cases were at L1-2,8 at L2-3,12 at L3-4,186 at L4-5 and 141 at L5S1.The automatic nerve retractor was used in all micro-endoscopic discectomy.The visual analogue scale (VAS) and Oswestry disability index (ODI) were used to evaluate clinical outcomes.Results All patients were followed up for 6 to 16 months (average,9 months).The mean VAS score decreased from preoperative 8.79±1.15 to 3.80±1.14 3 months after operation and 3.65±1.14 6 months after operation.The mean ODI score decreased from preoperative 78％±1.71％ to 28％±1.72％ 3 months after operation and 28％±1.88％ 6 months after operation.Postoperative VAS and ODI scores decreased significantly compared with those before operation.The VAS and ODI scores 6 months after operation were not significantly improved compared with those 3 months after operation.No spinal cord and nerve root injury and epidural hematoma formation occurred in all cases.Conclusion In micro-endoscopic discectomy,the automatic nerve retractor can help the operator obtain effective exposure,protect nerves from injury,alleviate workload of the operators,therefore it has a great clinical application value.

BACKGROUND:Transplantation of mesenchymal stem cels to prevent and treat degeneration of the intervertebral disc is a feasible method. Mesenchymal stem cels co-transfected by SRY-related high mobility group-box gene 9 (SOX-9) and growth differentiation factor-5 (GDF-5) can differentiate into nucleus pulposus cels, in order to obtain greater effect of induction and proliferation of nucleus pulposus cels. OBJECTIVE:To investigate the effect of SOX-9 and GDF-5 co-transfection on the differentiation of rabbit bone marrow mesenchymal stem cels into nucleus pulposus cels. METHODS: We separated and cultured bone marrow mesenchymal stem cels from the bone marrow of rabbit aged 4 months. Passage 3 cels were divided into five groups andin vitro induced to differentiate into nucleus pulposus cels: non-transfected group, empty vector transfection group, SOX-9 transfection group, GDF-5 transfection group, SOX-9 and GDF-5 co-transfection group. At 14 days after transfection, RT-PCR was employed to assay SOX-9, GDF-5 and colagen type II mRNA expressions in bone marrow mesenchymal stem cels. The marker of nucleus pulposus cels-KRT19 expression was also detected by immunohistochemical staining. RESULTS AND CONCLUSION:In the co-transfection group, the mRNA expressions of SOX-9, GDF-5, and colagen type II were significantly higher than those in the SOX-9 transfection group, GDF-5 transfection group, and both these two groups, respectively (P < 0.05). Cels were positive for KRT19 in the SOX-9 and GDF-5 groups, and strongly positive for KRT19 in the co-transfection group. These findings indicate that double gene-transfected bone marrow mesenchymal stem cels are better than single gene-transfected cels with regard to differentiation into nucleus pulposus cels and secretion of extracelular matrix.