Objective To observe the difference in anticoagulant ability of endothelial cells from different sources.Methods Bone marrow mesenchymal stem cells(BMMSCs)were cultured,purified,and expanded by Ficoll-Paque density gradient centrifugation and adherent culture in vitro.Then they were induced and differentiated in medium with 10 ?g/L VEGF.After 7 days,Von Willebrand factor(VWF)of the cells were identified by immunohistochemistry.At last,the major anticoagulant gene expression of the vascular endothelial-like cells derived from BMMSCs and the human umbilical vein endothelial cells(HUVECs)was detected and compared by reverse transcriptase PCR(RT-PCR).Results Though BMMSCs can successfully differentiate into vascular endothelial-like cells in vitro,they fail to express mRNA of the major anticoagulant gene.However,HUVECs can express the mRNA of these genes.Conclusion BMMSCs can differentiate into vascular endothelial-like cells in vitro,but their anticoagulant ability is inferior to HUVECs.

AIM:Repair of trachea is disturbing the surgeon. Tissue engineering technology will probably resolve this problem. Seed cell is one of the key factors in engineered tracheal cartilage construction. This study investigated the feasibility of constructing tissue-engineered cartilage from porcine bone marrow mesenchymal stem cells(MSCs) cultured and induced in vitro using tissue engineering technique. METHODS:The experiment was performed in the Central Laboratory of Peking Union Medical College Hospital between October 2006 and May 2007. ①By density gradient centrifugation,the MSCs were isolated and purified from porcine bone marrow. The MSCs had been cultured and induced in the defined medium mainly including transforming growth factor-?1,and then the type Ⅱ collagens were detected by immunohistochemical assay. The induced MSCs were seeded onto polyglycolic acids(PGA) scaffold as experimental group,and PGA scaffold were implanted into subcutaneous tissue as control group. The cell-scaffold construct was wrapped around a silicon tube(0.4 cm in diameter) and implanted into subcutaneous tissue of porcine. All specimens were harvested after in vivo culture for 6,8 and 10 weeks and evaluated by gross view,histology,and immunohistochemistry. RESULTS:①The MSCs were obtained by density gradient centrifugation method,and abundant seed cells were obtained after culture and amplification. ②The MSCs differentiated towards chondrocyte when cultured in the specific medium in vitro and were verified by the positive result of collagen type Ⅱ through immunohistochemistry. ③After implanted into subcutaneous tissue for 6,8 and 10 weeks,the cell-scaffold formed a tubular cartilage,which was very similar to normal porcine tracheal cartilage in both gross view and histology. And the result of collagen type Ⅱ through immunohistochemistry was positive. CONCLUSION:The in vivo and vitro cultured MSCs from porcine bone marrow can generate tissue-engineered cartilage under chondrogenic induction.

Objective The polysaccharides sulfate were extracted from three kinds of algae in-cluding porphyra tenera , Laminaria ja ponica and Sargassum fusi forme ( Harv. )Setch . in order to screen antioxidant with exploiting potential. Methods Using the assay system of DP-PH,the antioxidative activities of various extracts were studied. Results Three kinds of algae polysaccharides sulfate had different antioxidative activities. Moreover, polysaccharide extracted with different solvents had different intensities of antioxidative activity. Conclusion Polysacchride sulfate extracted with water has stronger antioxidative activity than that with others.keywolds antioxidation; porphyra tenera kjellm. ; Laminaria japonica; Sargassum fusi -forme ( Harv. )Setch.Studies on the extraction of polysaccharides sulfate from three algae and their scavenging activity on free radicalsHAN Hua,ZHOU Hai Yan,LIU Cheng-Chu,WANG Chun-bo(1.Bromatology Institute of Shanhai Aquatic University , Shanghai 200090, China; 2. Medical College , Qingdao University ,Qingdao 266021, China)Abstract:Objective The polysaccharides sulfate were extracted from three kinds of algae in-cluding porphyra tenera , Laminaria ja ponica and Sargassum fusi forme ( Harv. )Setch . in order to screen antioxidant with exploiting potential. Methods Using the assay system of DP-PH,the antioxidative activities of various extracts were studied. Results Three kinds of algae polysaccharides sulfate had different antioxidative activities. Moreover, polysaccharide extracted with different solvents had different intensities of antioxidative activity. Conclusion Polysacchride sulfate extracted with water has stronger antioxidative activity than that with others.

Objective Study on the diagnosis value of expression of serum macrophage inhibitory factor 1 (MIC-1) combined with alpha-fetoprotein isoforms 3 (AFP-L3) in primary liver cancer detected by enzyme linked immunosorbent assay (ELISA).Methods MIC-1 and AFP-L3 concentrations were detected by ELISA from selected 116 patients of primary liver cancer.Results were compared both in combined and sigle detection.Results In primary liver cancer group AFP-L3 concentration [(127.12±51.43) ngmml] was significantly higher than that in normal control group [(27.11±7.26) ng/ml,P ＜ 0.001].With AFP-L3 ＞ 38.0 ng/ml as the critical value,the sensitivity was 85.34 ％ (99/116),specificity was 88.33 ％ (53/60) and the diagnostic accuracy was 86.36 ％ (152/176).In primary liver cancer group MIC-1 concentration [(3140.43±1138.23) pg/ml]was significantly higher than that in normal the control group [(701.88±302.34) pg/ml,P ＜ 0.001],the sensitivity was 91.38 ％ (106/116),specificity was 85.00 ％ (51/60),the diagnostic accuracy was 89.20 ％(157/176).The two combined detection sensitivity was 83.62 ％ (97/116),specificity was 91.67 ％ (55/60),diagnostic accuracy was 86.36 ％ (152/176).Conclusion MIC-1 combined with AFP-L3 concentration detection can improve the specificity of the diagnosis of primary liver cancer,which has certain clinical value.

Animals ; Male ; NF-kappa B ; metabolism ; Otitis Media with Effusion ; immunology ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Th1 Cells ; cytology ; Th2 Cells ; cytology

ObjectiveTo investigate the effect of clinical pathway teaching methord in nursing practice teaching. Methods80 junior college nursing students were randomly divided into control and experimental groups. Traditional clinical teaching method was given to control group, while the clinical pathway teaching method was given to observation group. Scores of comprehensive quality after departmental rotation and satisfaction rates of nursing students to teaching method in these two groups were evaluated. ResultsThe experimental group was significantly better than the control group ( P＜0.05 ), and the difference was statistically significant. ConclusionThe clinical pathway can significantly improve the quality of nursing practice teaching.

By using electron microscopy, the paranodal region and axo-glial junction were examined in optic nerves of rats aged 14 days. The paranodal region was characterized in longitudinal sections by the sequential termination of the myelin lamellae, beginning proximally with the innermost and ending, at the Ranvier node, with the outermost lamella. The termination of each lamella was accom- panied by a separation of the major dense line of the compact myelin and the consequent formation of a "loop" of glial cytoplasm. Each paranodal loop inde- nted the axonal surface as it became junctionally apposed to the axolemma. The periaxonal extracellular space, 10-20nm in width in the internodal region and reduced at the paranodal junction to approximately 3nm, forming an axo-glial junction, which was thought to be held together by dense structure. The parano- dal junction seems to serve strong adhesion between the apposed axonal and glial membranes. Conduction of the nerve impulses in myelinated axons was saltatory. Axons and sheath cells probably maintain vital communication with one another, presumably at the paranodal junctional complex. This communication was viewed as vital to the stability and maintenance of myelin. We found some clear vesi- cles in axoplasm near the Ranvier node and speculated that there were endocyto- sis and exocytosis in paranodal region. This was a direct morphological evidence supporting metabolic coupling between axons and sheeth cells.

Objective To study any effect of repetitive magnetic stimulation (rMS) on the differentiation and apoptosis of rat neural stem cells in vitro.Methods The bilateral hippocampus of a 3-day old Sprague-Dawley rat was used to culture neural stem cells (NSCs) in vitro.P2 NSCs were differentiated to neurons or astrocytes in differentiation medium and then divided into a control group in which the NSCs differentiated naturally,and an rMS group in which 1000 impulses/day of rMS were applied at 10 Hz once a day for 7 days at 50％ of maximum output.One hour after the last stimulation,immunofluorescence was used to analyze the ratio of neurons and astrocytes,and Western blotting was employed to evaluate the expression of glial fibrillary acidic protein (GFAP),β-Ⅲ tubulin and brain-derived neurotrophic factor (BDNF).NSCs which had differentiated for 7 days without stimulation were then selected and divided into an apoptosis group and an apoptosis+rMS group.The same rMS protocol was applied to the latter group 1h after the apoptosis,and 4h later flow cytometry (anexin V-FITC) was employed to evaluate the apoptosis ratio.Bcl-2,Bax and caspase-3 protein expression were analyzed using Western blotting.Results There were no significant differences between the control and rMS groups in the proportion of NSCs differentiating to neurons or in β-Ⅲ tubulin,GFAP or BDNF protein expression.The cell apoptosis rate of the apoptosis+rMS group was significant lower than in the apoptosis group.Caspase-3,Bcl-2 and Bax protein expression were also significantly different between the two groups.Conclusion rMS at 10Hz for 7 days has no effect on the differentiation of NSCs,but it has a protective effect on neural cells and decreases the apoptosis rate.

Smoothened (SMO) is a member of sonic hedgehog homology (SHH) signaling pathway. It plays a key role as a bridge between patched-1 (PTCH-1) and Gli. Aberrant SHH expression can be detected in various malignant tissues, and the expression in pancreatic cancer stem cells is higher apparently. SHH signals are closely associated with self-duplication of cancer stem cells, formation of tumor vessels as well as matrixes. SMO antagonists such as cyclopamine, GDC-0449 and so on show potential to inhibit activity of SHH signaling, and arrest the growth as well as metastases of tumors. Recently, a few of SMO antagonists have been studied in phase I clinical trials and some are in phase II, meanwhile, phase I or II trials of SMO antagonists to treat pancreatic cancer are performed currently. As the classical SMO antagonist, cyclopamine is extracted from a medicinal plant. Perhaps researchers may be able to determine more effective SMO-targeting drugs from herbal medicines in the future.

Objective To explore the perioperative nursing of patients from high altitude earthquake-stricken regions. Method The perioperative nursing histories of 18 patients from high altitude earthquake-stricken regions were retrospectively analyzed to summarize the nursing experience.Results No wound infection,crush injury or complications occurred.Sixteen patients had good functional recovery,1 patient was at the stage of recovery about limb movement and another one was also at the recovery period from lumbar vertebral burst fracture complicated with nerve injury.Conclusion Close observation,psychological nursing and functional exercise are important for improved success rate of treatment for the patients from the earthquake-stricken regions.