Objective:To study the expression of kringle 1-3 domain fragment of human plasmihogen in E.coli and the anti-tumor activity of its product.Methods:The K1-3 domain fragment was cloned in expression vector pBV220,the resulted recombinant plasmid pBV-K13 was transformed into E.coli DH5? and its product was purified and assayed its bioactivity.Results:K1-3 domain fragment was expressed in(E.coli) DH5?.The results showed the expressed product covered 20% of the total bacterial protein on SDS-PAGE and the Western blot analysis showed that the product had immunological specificity with the antiserum of human plasminogen and inhibits the growth of chorioallantoic membrane(CAM) angiogenesis and mouse B16 melanoma.Conclusion:Human plasminogen K1-3 domain fragment was expressed in E.coli;the expressed product has anti-angiogenesis and anti-tumor activity.

Objective To investigate different frequencies of glucocorticoids (GCs) on the tissue in a model of osteoporosis. Methods Thirtytwo three-month-old SD female rats were randomly divided into four groups: (1) the control group (group C); (2) the low-frequency group (group L); (3) the middle-frequency group (group M); (4)the high-frequency group (H). The rats in the group C were given intramuscular injection (im) of 0.9% saline. Im of dexamethasone (Dex) was 1 mg/(kg·time). Rats were given two times im a week in the group C, four times im a week in the group M, and six times im a week in the group H. Each rat was sacrificed on thirty days post-administration. Results (1)The body weight of rats gradually increased in the Ctrl group , however , the body weight of rats declined gradually during the experiment in the group L, M and H. The size of immune organs (spleen and thymus) significantly decreased in rats of the group L, M and H. (2)Compared with the group C, cell edema was changed in the heart, renal and lung morphological fatty degeneration in liver , atrophy in spleen , atrophy in lymphoid nodules , and cell edema in kidney tubular were observed. Conclusion GCs cause serious degradation in the thymus and atrophy of the spleen. Administration has different inhibitory effect on immune function; the high frequency will lead to strong inhibition.

Objective To study the inhibition of berberine on organ anion transporters (OATs) and its bidirectional trans-membrane transport.Method The transgene cell lines of the organ anion transporters including OAT1,OAT2,OAT3,OAT4,OAT7,and URAT1 were constructed and selected by animal cell transgenic method mediated by transporter Lipo 3000.Wild type (WT) cells were used as control group,and activity of OATs was verified by adding their radiolabeled substrates and inhibitors.The inhibition of 100 μmol/L berberine on the transporters was investigated in vitro.The IC50 of berberine on URAT1 was also determined.The bidirectional transport of berberine was studied through the Caco-2 model.Result The results showed that 100 μmol/L berberine inhibited the activity of OAT1,OAT2,OAT3,OAT4,OAT7 and URAT1 to (70.48±4.23)％,(69.13±1.28)％,(72.12±3.28)％,(79.77±6.49)％,(69.51 ±5.99)％ and (38.4 ± 2.67)％ respectively,the IC50 of berberine to URAT 1 was 13.19 μmol/L,the Papp (A-B) of 50 μmol/L and 100 μmol/L berberine were separately 0.28 × 10-6 and 0.40 × 10-6 cm/s,and the effiux rates were separately 3.18 and 3.15.Conclusion Berberine shows a stronger inhibition to URAT1 compared to OAT1,OAT2,OAT3,OAT4 and OAT7.Berberine may be the substrate of some effiux transporters.This study provides theoretical basis for explaining the low bioavailability ofberberine and forecasting the possible drug-drug interaction.

Objective To investigate the influence of polyethylene glycol (PEG) on heparin release of heparinloaded polycaprolactone/polyethylene glycol (PCL/PEG) membranes used in artificial vascular peosthesis.Methods Heparin-loaded PCL/PEG membrane samples with different PEG mass contents of 0,0.5％,10％ and 15％ were prepared by blending method and freeze-drying technology.The influence of PEG on heparin release was experimental studied in vitro.The influence of PEG on the structural characteristics of the samples were investigated by X-ray diffraction,Fourier transform infrared spectrum and differential scanning calorimeter.Results The addition of PEG reduced the heparin release resistance.The results showed that the average release rate of heparin in the first day and the release amount for 34 d were improved.Both these parameters increased with the increase of PEG mass content.The X-ray diffraction,Fourier transform infrared spectroscopy and differential scanning calorimetry showed that the crystallinity of PCL membrane was slightly enhanced by the addition of heparin,but the overall effect was not significant.In addition,the addition of heparin could promote the crystalline grain growth of PEG,and a common distribution of heparin and PEG in the matrix was observed.Conclusions The heparin release control can be achieved by adjusting the PEG mass content in heparin-loading PCL/PEG membranes prepared by blending method and freeze-drying technology.The proposed samples may have anticoagulant effect,which can be expected to be used as small-diameter artificial vascular prosthesis material.

Aim To study the influence of Sodium fer-ulate ( SF) on bone metabolism in glucocorticoid–in-duced osteoporosis rats. Methods Thirty cases of fe-male Wistar Rats(3-month-old) were divided into con-trol group, model group and SF group ( low-dose group, middle-dose group, high-dose group ) by ran-domized block design. Double fluorochrome labeling with calcein was performed before necropsy. The left tibia was taken for bone histomorphometry. Results In static parameters, the proximal tibia cancellous bone trabecular thickness, trabecular quantity and area ratio were significantly reduced in model group compared with control group;while compared with model group, those were increased in middle and high-dose SF group. Trabecular separation degree was increased in model group compared with control group, while it was decreased in middle and high-dose SF group compared with model group. In dynamic parameters, the calcula-tion parameters of cancellous bone mark perimeter rate and the bone formation rate were increased in model group compared with control group, in middle and high-dose SF group the bone formation rate was in-creased compared with model group. In bone cells, os-teoclast number per mm, osteoblast number per mm, percent osteoblast surface perimeter and percent osteo-clast surface perimeter were increased in model group compared with control group. In growth-plate, the thickness of growth-plate was increased in model group compared with control group. In bone cells and growth-plate there was no statistical significance between treat-ment group and model group. Conclusion This study demonstrates that SF can increase bone mass and im-prove bone structure,which may be related to the im-provement of bone formation. SF is effective for GIOP in rats.

Objective To study the inhibitory effects ofberberine on human organic cation transporter (OCTs) including OCT1,OCT2,OCT3,OCTN1 and OCTN2.Methods Using animal cell transgenic method mediated by transporter Lipo 3000,the drug transporters over expression cell lines S2-OCT1,S2-OCT2,S2-OCT3,S2-OCTN1 and S2-OCTN2 were obtained by selective medium culture.The OCTs evaluation model was established by detecting the trans-membrane transport of radioactive substrate in vitro.Wild type (WT) cells were used as control group,activity of OCTs was verified by adding its inhibitor.The inhibition of berberine on the transporters was investigated in vitro.The IC50 of inhibitory effect of berberine on various drug transporters was also calculated.Result The transport activity of transporter cell lines was increased by more than 5 times compared to the WT cell line respectively,what's more,their transport activity decreased significantly by their corresponding inhibitor.The ICs0 of berberine to OCT1,OCT2,OCT3,OCTN1 and OCTN2 were respectively 7.63,6.80,2.25,4.66 and 210.34 μmol/L.Conclusion Berberine significant inhibition to OCT1,OCT2,OCT3,OCTN1 and OCTN2.The inhibition on OCT1,OCT2,OCT3,OCTN1 is stronger compared to OCTN2.