Objective:This experiment aims to find a appropriate scaffold for Tissue Engineer- ing.Methods:The chondrocytes were cultured when they were seeded onto PGA+PLA scaffolds coat- ing with Lecithin(LEC)and Poly-l-lysine(PLYS)together and respectively.With light microscope and scanning electron microscope,the observation of the scaffolds in hydrophilia and adsorptivity to chondrocytes and the function of the cells was made.Results:The PGA scaffolds coating with LEC and PLYS have better hydrophilia and adsorptivity to the cells;on which the chodrocytes produce more ma- trices.Conclusion:LEC can chang the hydrophilia of the scaffolds;while PLYS can strengthen the ad- sorptivity of the scaffolds;the PGA coating with LEC and PLYS is an ideal scaffold in Tissue Engineer- ing.

Adult ; Aged ; Arteriosclerosis Obliterans ; drug therapy ; Asteraceae ; chemistry ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Female ; Heparin ; therapeutic use ; Humans ; Infusions, Intravenous ; Leg ; blood supply ; Male ; Middle Aged ; Phytotherapy

AIMTo detect the expression of a novel protein, hemangiopoietin (HAPO), in the human fetal liver at the protein level.

METHODSMonoclonal antibodies (moAbs) against HAPO were produced by traditional hybridoma technique. Their affinities were calculated from the results of non-competitive ELISA and the antibodies were purified by protein G affinity chromatography. Expression of HAPO in the liver was detected by SDS-PAGE and Western blot.

RESULTSFive strains of moAbs were screened out in total, among which three were IgG1 and the other two were IgM. Their light trains were all belonged to kappa. The relative affinities of the three IgG1 form moAbs were 3.06 x 10(9) mol/L, 6.07 x 10(8) mol/L and 1.71 x 10(10) mol/L respectively. After purification, the purity of the moAbs could reach more than 99%. HAPO expression was detected at the protein level in the human fetal liver, and the apparent molecular weight of the nature HAPO was very close to but a little higher than our recombinant one.

CONCLUSIONHAPO was expressed in the human fetal liver at the protein level.

Antibodies, Monoclonal ; Blotting, Western ; Fetus ; Humans ; Immunoblotting ; Liver ; embryology ; metabolism ; Proteoglycans ; metabolism

Objective: To construct an expression system containing the N-terminal segment of Thrombospondin-1 (TSP-1) in E. coli. To investigate the activity of TSF. Methods: thbs1 gene fragment was amplified from human fetal cord vein endothelial cells and inserted into plasmid pET32c ( + ) which was then transfected and expressed in E. coli. Investigate the effect of recombinant TSF to the proliferation of ECV304 in vitro with MTT. The expression level of CD36 was assayed by FACS. Results: Recombinant TSF was purified. TSF could restrain the proliferation of ECV304 with CD36 low-expression. Conclusions:Low dose of rTSF was a latent assistant treatment of anti-cancer.

The diagnosis,surgical treatment,and comprehensive treatment of renal cell carcinoma with inferior vena cava tumor thrombus have advanced rapidly in recent years. Both the survival and quality of life of the patients have remarkably improved. Further advance in basic research may provide new direction of management of renal cell carcinoma.
Carcinoma, Renal Cell ; complications ; diagnosis ; therapy ; Embolism ; diagnosis ; etiology ; therapy ; Humans ; Kidney Neoplasms ; complications ; diagnosis ; therapy ; Venae Cavae

OBJECTIVETo study the effect of PF4 on the adherence of leukemic CD34+ KG1a cell to human umbilical vein endothelial cell line ECV-304 cell and on the expression of adhesive molecules.

METHODSAdhesion assay and adhesion blocking assay were respectively applied to measure the effect of PF4 and/or adhesion molecule monoclonal antibodies on the adhesion property of KG1a. The expressions of adhesion molecules were determined by RT-PCR and FACS analysis.

RESULTSThe adhesion of KG1a cells to ECV-304 was significantly enhanced in the presence of PF4. Such enhancement was also observed when KG1a or ECV-304 cells were separately treated with PF4 before interaction. The adhesion capacity of KG1a cells was reduced when cells were co-incubated with the blocking monoclonal antibodies (MoAbs) against CD49d, CD106, CD54, respectively. In contrast, MoAbs against CD62L, CD62P and CD62E had no such effect. During a period of 3 hours when KG1a or ECV-304 cells were respectively incubated with PF4, the mRNA expressions of CD49 d, CD54 were up-regulated. Furthermore, when KG1a or ECV-304 cells were incubated with PF4 for 2 hours, respectively, the percentages of CD49d+ KG1a cells and CD54+ ECV-304 were increased significantly.

CONCLUSIONPF4 can enhance KG1a cell adhesive capacity by increasing the expressions of adhesion molecules.

Antigens, CD34 ; metabolism ; Cell Adhesion ; drug effects ; Cell Adhesion Molecules ; metabolism ; Cell Line, Tumor ; Humans ; Leukemia, Myeloid, Acute ; immunology ; pathology ; Platelet Factor 4 ; pharmacology ; Umbilical Veins ; cytology

Hematopoietic stem cell (HSC) is the first discovered and well studied tissue stem cell. HSC transplantation has been successfully applied to cure a variety of diseases of hematological and immunological systems. It has long believed that HSC and angioblast come from the common stem cell, the hemangioblast. Recently, HSC has been demonstrated to be able to differentiate into vascular endothelial cells. In animal in vivo models, HSC transplantation can promote angiogenesis and improve limb ischemia. Several pilot clinical studies have shown that transplantation of bone marrow and granulocyte colony stimulating factor mobilized peripheral blood HSC promoted vascular reconstitution in ischemic limbs. Umbilical cord blood has been an important source of HSC for clinical transplantation. Animal studies have demonstrated the efficiency of cord blood HSC transplantation in improving critical limb ischemia. These studies have provided evidences that HSC can be used for the treatment of vascular diseases.
Animals ; Cell Differentiation ; Extremities ; blood supply ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic Stem Cells ; cytology ; Humans ; Ischemia ; therapy

To construct a scFv library by phage display technique from the spleen cells of mice immunized with B3HM cells. Three mice were immunized with B3HM cells, and their spleen cells were harvested. The genes of VH and Vk were amplified by RT-PCR from the cDNA of the immunized spleen cells and a scFv-phage display antibody library was constructed. The capacity of library was measured,and the variety of the library was analyzed by digesting with restriction endonuclease BstNI.ScFv phage clones were randomly picked and identified phage-scFv clone by binding B3HM cells using immunofluorescein.A scFv library containing 5?106 individual clones which showed different patterns after digested with restriction endonuclease BstNI was produced. Individnal phage-scFv clone showed B3HM cells positive using immunofluorescein. A scFv library of anti-B3HM cell surface molecules has been constructed. It will be useful for finding out some novel genes of causing leukemia, and establishs the infarctate foundation of clarifying the pathogenesis of leukemiagenesis.

Objective To investigate the differential expression of P57kip2 and CDK5 in neural tube defects t(NTD) from the normal ,and provide the clue for the research of the molecular mechanism of the normal neurula formation .Methods A cDNA mi-croarray containing 1 100 known genes was used to compare differences in P57kip2 and CDK5 gene expression between the normal control group and the retinoic acid(RA)-induced NTD group on embryonic(E) day 9 .5 and 10 .5 .Two differentially expressed genes were randomly selected from the two groups for Northern blotting to verify the results of the cDNA microarray .Results Compared the differences of between P57kip2 and CDK5 in normal and E9 .5 d ,E10 .5 d ,E9 .5 d-NTD ,E10 .5 d-NTD ,P57kip2 and CDK5 expression was significantly up-regulated in the before and after the formation of the normal neurulation ,but them showed a downward trend in retinoic acid (RA)-induced NTD(including two phase E9 .5 d and E10 .5 d) .Conclusion P57kip2 and CDK5 in-volved in the physiological process of NTD ,and provide the useful clue for the research of the molecular mechanism of the normal neurula formation .

Objective To explore the clinical value of ureteroscopy and minimally invasive percutaneous nephrolithotomy ( MPCNL) in the treatment of ureteral calculi patients with acute renal dysfunction .Methods Clini-cal data of 124 ureteral calculi patients with acute renal dysfunction were retrospectively analyzed .86 cases were trea-ted with holmium-laser lithotripsy under ureteroscope .38 cases were treated with MPCNL under the guide of ultra-sound.Results Three months after operation ,the stone clearance rate was 100%,and no severe complications were observed.The renal function decreased to normal levels in 102 cases(82.3%).Conclusion The holmium laser lith-otripsy under ureteroscope and MPCNL can deal with double sites of ureteral calculi ,and offer advantages with less in-vasion,safety and efficiency ,which can be the first choice for the ureteral calculi combined with acute renal dysfunc -tion.