Objective: To investigate the effects of different concentrations of vitamin E (VE) on angiogenesis and its possible mechanism. Methods: The rings of rat aorta were embedded in gels of collagen and cultured in a serum free medium for 28 days. Curves of microvessels growth were generated by counting the number of newly formed microvessels every day with an inverted microscope. Photos were taken at the same time. Expression of factor FVIII related antigen (FVIII RAg) in newly formed microvessels was detected by immunohistochemistry. Concentration change of FVIII R in medium was evaluated by ELISA. Results: Both 0.2 g/L and 0.1 g/L VE showed significant inhibitory effect on angiogenesis (P0.05). The inhibitory effect of VE was expressed in the phases of microvessel growth and decline (10-25 d). Conclusion: 1.Three dimensional collagen gels culture of microvessel in serum free medium can be used as a sensitive assay for study of soluble and solid phase angiogenic agonists and antagonists. 2. Both 0.1 g/L and 0.2 g/L VE inhibit angiogenesis, and VE has no toxicity and no drug resistance.

From the development of national clinical research base of traditional Chinese medicine ( TCM ) in Longhua Hospital, Shanghai University of Traditional Chinese Medicine, we made a success of construction, such as planning and foundation , curative effect , study protocol , standards , platform establishment . From the thinking to results , we made knowledge of the situation in order to provide references for the further building of clinical research base of TCM .

Objective To isolate and culture human adipose-derived stem cells (hADSCs),investigate the influence of hADSCs on the cellular morphology,survival rate,and function of human islet cells under the in vitro non-contact co-culture conditions,and explore its mechanism.Method hADSCs were isolated by collagenase digestion method,then cultured,and identified by morphology,immunofluorescence and multi-directional differentiation.Adult islet cells were separated and purified by Liberase enzyme and Ficoll 400,then divided into co-culture group and individual group.The cellular growth morphology of islet cells was observed by inverted phase contrast microscope.The survival rate of islet cells,insulin secretory volume,insulin stimulation index and concentration of growth factor in the supernatant were compared between the two groups.Result hADSCs of the third generation showed uniform long spindle fibrocyte-like morphology,and had multi-directional differentiational potentials of osteogenesis and adipogenesis.Immunofluorescence test of surface antigens on hADSCs revealed CD44 + and CD49d +,CD31-,CD34-and CD106-.After 14-day culture,the islet cellular morphology in co-culture group was more intact than that in individual group.The survival rate of islet cells in co-culture group was (82.83 + 2.32) ％,and that in individual group was (53.00 + 2.82) ％ (P＜0.01).Insulin secretory volumes were (23.66 + 2.11) and (7.82 +1.09) mU/L respectively in co-culture group and individual group under high glucose concentration,and 13.22 + 0.77 and 6.40 + 0.44 mU/L respectively under low glucose concentration (P＜0.01 for all).Insulin stimulation index was decreased from 1.67 + 0.10 (at 3rd day) to 1.77 + 0.13 (at 14th day) in co-culture group,and from (1.67 + 0.10) (at 3rd day) to (1.77 + 0.13) (at 14th day) in individual group (P＜0.01).After 14-day culture,the concentrations of HGF,TGF-β,VEGF and bFGF in the supernatant were higher in co-culture group than in individual group (P＜0.01).Conclusion hADSCs were isolated and cultured successfully from adult adipose tissue.They could increase the survival rate and improve the function of islet cells when co-culture with the adult islet cells in vitro through secreting HGF,TGF-β,VEGF and b-FGF.

ABSTRACT:Objective For preparation of vascularized islets , to isolate and culture human adipose derived stem cells , investigate the role of adipose derived stem cells (ADSCs ) in promoting the proliferation and vascularization of human umbilical vein endothelial cells (HUVECs ) co‐cultured in vitro , and explore its mechanism .Methods ADSCs and HUVECs were isolated by collagenase digestion method ,then cultured ,and identified by morphology ,immunofluorescence or multi‐directional differentiation .The co‐culture system of ADSCs and HUVECs was established , HUVECs cultured alone were set up for control group . The proliferation , vascularization and concentration of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b‐FGF)in the supernatant were compared between the two groups .Results The third generational ADSCs had uniform long spindle fiberous morphology and multi‐directional differentiational function . Immunofluorescence test of surface antigens on ADSCs revealed CD44/CD49d (+ ) ,CD31/CD34 (-) ,on HUVECs CD31/vWF (+ ) . High vascular density was found when co‐cultured in Matrigel of ADSCs and HUVECs than alone of HUVECs .Growth curve shown at days 3 , 4 and 5 of the logarithmic phase , HUVECs count in co‐culture group of ADSCs and HUVECs was (4 .52 ± 0 .31) × 104 ,(7 .18 ± 0 .45) × 104 ,and (8 .23 ± 0 .36) × 104 under indirect co‐culture condition , while that in individual HUVECs group was (2 .71 ± 0 .25) × 104 ,(4 .87 ± 0 .26) × 104 ,and (6 .86 ± 0 .33) × 104 ( P<0 .01) .Population doubling time of HUVECs was shorter in co‐culture group than in individual group .Also ,the OD value of HUVECs was higher in co‐culture group than in individual group when cultured at days 1 ,3 ,5 and 7 ( P<0 .01) .When cultured at days 3 ,7 and 13 ,the concentration of VEGF and b‐FGF in the supernatant was higher in co‐culture group than in individual group ( P< 0 .01 ) . Conclusion ADSCs can promote the proliferation and vascularization of HUVECs in vitro co‐culture conditions by secreting or increasing the HUVECs secretion of VEGF and b‐FGF .

ObjectiveTo determine thallium in whole blood by atomic absorption detection method, and to investigate the eliminating effect of hemoperfusion (HP) for thallium in blood.Methods The blood of Beagle dogs which had not exposed to thallium before were obtained for preparation of thallium nitrate (TlNO3)-containing solution in three concentrations according to the conversion formula based on animal weight and volume of blood. HP was performed in the simulated in vivo environment. The content of TlNO3 in blood of the next group was determined on the amount of TlNO3 for the last HP of the former dose group. Thallium quantity in different samples was measured with atomic absorption spectrometer blood samples before and after HP. Finally, the thallium concentration in blood was analyzed statistically.Results Thallium concentrations showed a good linear relationship in the range of 0-200μg/L (r = 0.998 4). The intra-day precision (RSD) was lower than 4.913%, the intra-day recovery rate was 96.2%-111.9%; the inter-day precision (RSD) was lower than 7.502%, the inter-day recovery rate was 89.6%-105.2%. The concentration of thallium in blood was significantly reduced after HP per time in high, middle, and low dose groups [(453.43±27.80) mg/L to (56.09±14.44) mg/L in high dose group,F = 8.820,P = 0.003;(64.51±13.60) mg/L to (3.19±0.23) mg/L in middle dose group,F = 36.312,P = 0.000; (5.40±0.98) mg/L to (0.38±0.25) mg/L in low dose group,F = 46.240,P = 0.000]. The adsorption rate of four times of HP in high, middle and low dose group were (87.63±2.48)%, (95.06±1.54)% and (92.76±4.87)%, respectively, without significant difference (F = 4.231,P = 0.070 ).Conclusions The method for measuring thallium was established, and it shows a very stable, simple, sensitive for determination of thallium. HP can effectively remove thallium from blood. Thallium concentration can be reduced by 90% after four times of HP. HP is also effective even when thallium concentration is not high.

Objective: To establish a method for the determination of furans in tea by headspace-gas chromatographic mass spectrometry. Methods: The 20% sodium chloride solution and isotope internal standards were added to the crushed and weighed tea sample. Furan, 2-methylfuran, 3-methylfuran, 2,5-dimethylfuran were separated by HP-PLOT Q capillary column and then determined by gas chromatography mass spectrometry with electron impact ionization mode. Results: In the range of 5-400 ng, good linear relationships were observed in the four furan compounds, with the correlation coefficients ranging from 0.999 2 to 0.999 6. The detection limits ranged from 0.2 to 1.9 μg/kg, the quantification limit ranged from 0.4 to 3.1 μg/kg. The recovery rates of furans ranged from 95.4% to 128.2% when spiked at 5.0, 20.0 and 100.0 μg/kg, and the relative standard deviations ranged from 0.8% to 11.3%. Eighty-one tea samples were determined, the concentration of four furan compounds was highest in black tea, followed by dark tea, oolong tea, green tea and scented tea. Conclusion Headspace-gas chromatography-mass spectrometry can reduce the matrix interference of tea, and meet the requirements in the linear range, recovery and precision, which is suitable for simultaneous determination of four furan compounds in tea.

Objective: To investigate the effects of psoralen and angelicin on inflammation cytokine expression of human periodontal ligament cells (hPDLCs). Methods: hPDLCs were primarily cultured using tissue explant method. Effects of psoralen and angelicin on the cell viability were tested by CCK-8 assay. hPDLCs were stimulated by Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) after treatment with different concentrations of psoralen and angelicin for 2 h. mRNA expression of IL-1β and IL-8 were determined by real-time PCR. Enzyme-linked immunosorbent assay (ELISA) was used to measure the secretion of IL-1β and IL-8. Results: hPDLCs were cultured successfully by tissue explant method. Psoralen and angelicin (≤ 12.5 μg/mL) did not show significant effects on the cell viability of hPDLCs. Pg-LPS markedly elevated the mRNA expression of IL-1β and IL-8, which could be attenuated by psoralen and angelicin in a dose-dependent manner. Likewise, the up-regulated protein secretion of IL-1β and IL-8 could be significantly blocked by psoralen and angelicin. Conclusion: Psoralen and angelicin could attenuate the inflammatory response of hPDLCs induced by Pg-LPS. Therefore, psoralen and angelicin may act as natural agents to prevent and treat periodontitis.

The secondary metabolites of plants are important sources of natural drugs. Betula plants have abundant pharmacological value, complex mechanism and wide applications, which are closely related to the triterpenoids of theirs. Triterpenoids in Betula species are mainly divided into dammarane-type, ocotillol-type, oleanane-type, lupane-type and cycloaltunane-type. The extracts of Betula species have varieties of activities such as anti-tumor, anti-inflammatory, anti-oxidant, anti-bacterial, etc. And the biosynthetic pathways of triterpenoids after 2,3-oxidosqualene are split into four branches of dammarenediol-II, lupeol, cycloartenol and amyrin according to the different oxidosqualene cyclases. This review summarizes the chemical constituents, pharmacological activities and biosynthetic pathways of triterpenoids in Betula plants. It provides a reference for the research and development of new drugs and the production of these triterpenoids in microbial cell factories by synthetic biology methods.

Objective To investigate the effects of asymmetric dimethylarginine (ADMA) on apoptosis and phosphorylation of c-jun N-terminal kinase (JNK) in endothelial outgrowth cells (EOCs). Methods The mononuclear cells were harvested from umbilical cord blood by ficoll density gradient centrifugation, and induced into EOCs and then expanded in vitro. The identified EOCs were treated with different concentrations of ADMA (0, 1, 5, 10, 20 μmol/L) for 48 h. The adherent cells were treated with 10 μmol/L ADMA,then different concentrations of JNK specific inhibitor SP600125 (0, 5,10,20 and 40 μmol/L) were added and incubated for 48 hours. Caspase-3 activity was measured by microplate reader. Apoptotic incidences of EOCs were quantitatively determined by flow cytometry. The expression of Caspase- 3 and phosphorylase-JNK (p-JNK) were detected by Western blot assay. Results The treatment of ADMA (1-20 μmol/L) significantly induced apoptosis in EOCs by enhancing Caspase-3 express and also induced phosphorylation of JNK (P<0.05). Meantime, the JNK specific inhibitor SP600125 could attenuate the apoptosis induced by ADMA during this process (F=6.733,P<0.05) and inhibit the expression of Caspase-3 and p-JNK. Conclusion ADMA can induce apoptosis in EOC, which may be achieved by activating JNK signal transduction pathway.

Objective To summarize the management of immature testicular teratomas in children.Methods The clinical data of 19 children (age between 16 days to 13 months) with immature testicular teratomas were retrospectively analyzed.There were 10 cases in left and 9 cases in right side.The main presentation was painless scrotal mass.The size of the tumor was 1.5 cm × 1.2 cm × 0.5 cm-6.0 cm × 5.0 cm × 4.5 cm.Abnormal alpha fetopmtein (AFP)concentrations were detected in 17 patients before surgery.Results Testis-sparing surgeries were done in 11 patients,and the other 8 patients underwent testicle resection because of tumor severely invading testis.Fifteen cases were followed up for 1-10 years.All of them were alive.Conclusions Immature testicular teratomas was rare and almost presenting in children younger than 1 year old.The biological behavior of immature testicular teratomas in children was benign.Testicular-sparing surgery was possible.Close follow-up should be done postoperatively.