Objective To observe the role of human hepatocarcinoma HepG2 cell culture supernatant on T lymphocyte of the activation, proliferation, IL-2 secretion and inducing apoptosis in vitro. To explore the mechanism of immune escape in tumor.Methods Peripheral blood mononuclear cells (PBMC) were cultured with different concentration of tumor cell culture supernatant. Cell proliferation was examined with standard cytometry and MTT. The apoptotic rate and immune phenotype were analyzed by FACS. The level of IL-2 was determined with ELISA.Results Adding HepG2 supernatant, there was no significant influence in activation of lymphocyte, but the proliferation rat and the secretion of IL-2 were obviously decreased. The lymphocyte apoptosis rate was raised.Conclusion The tumor cell may produce a certain soluble protein, thus escape from the immune surveillance of tumor-bearing host.

[Objective]To determine the content of Polygonum multiflorum thunb. Polysaccharide and the refined. [Methods] The sulfuric acid-phenol method was used. [Results] ?_ max = 486nm. The linear rang was 10.5～94.5?g?L -1 .And the average recovery was 101.23%(n=5,RSD=3.03%). The content of Polygonum multiflorum thunb. Polysaccharide was 14.28% and the refined was 19.03%. [Conclusion] The method was simple,rapid and reliable.

0.05),but the proliferation rate(P

[Objective]To investigate the possibility of using poly(hydroxybutyrate-co-hydroxyvalerate)(PHBV)as scaffolds for cartilage tissue engineering and to compare the engineered cartilage generated in vitro with those in nude mice modles.[Method]PHBV porous scaffolds were fabricated using a compression moulding,thermal processing and salt particulate leaching methods.Chondrocytes isolated from articular cartilage were seeded into porous PHBV scaffolds.After incubation for 2 weeks in vitro,chondrocytes-PHBV constructs were implanted subcutaneously in the dorsum of athymic nude mice.Control groups were established by subcutaneous implantation of PHBV alone.The implants harvested after in vivo incubation of 4 and 8 weeks and cell-scaffolds cultured in vitro for 6 and 10 weeks were respectively examined histologically.Chondrocytes cultured for 3,7 and 14 days on the scaffolds were examined by SEM.[Result]SEM showed that chondrocytes could aggregate and synthesize extracellular matrix on PHBV scaffolds.Both specimens harvested from nude mice and those cultured in vitro demonstrated new cartilage formation,while characteristics of the engineered cartilage generated in vivo were more typical.Those from control groups showed no cartilage formation.[Conclusion]PHBV can be used as scaffolds for cartilage tissue engineering and nude mice modle seems to facilitate chondrogenesis compared with that in vitro system.

Objective:To explore the role of CD28 in the activation of human peripheral blood ?? +T cells by Mycobacterium tuberculosis (Mtb) low molecular peptide antigen in vitro Methods:Mtb antigen and anti CD28 monoclonal antibody (mAb) were used as signal 2 to stimulate the r? + T cell from PBMC the expression of CD28 molecule on ?? +T cells, proliferation rate of ?? +T cells and expression of CD69 molecule on activated ?? +T cells were analyzed by using flow cytometry Results:CD28 molecule was expressed on 50% of ?? +T cells Neither Anti CD28 nor Mtb antigen alone, but both presence, could stimulate ?? +T cells activation and proliferation CD69 molecule was expressed on activated ?? + T cells.Conclusion:The CD28 molecule could provide the costimulatory signal in the full activation of ?? +T cells by Mtb low molecular peptide antigen CD69 molecule was also an early activation marker of ?? +T cells

Objective To investigate the influence of proprioceptive training of hemiplegic stroke patients on their visually compensatory dependence.Methods Twenty stroke patients without vestibular abnormalities were recruited.They were able to walk for 10 meters.All the patients underwent 4 weeks of rehabilitation training including proprioceptive training,treadmill exercises and isometric muscle strengthening.Static standing balance test was performed before and after 4 weeks of treatment.The parameters such as the anteroposterior and mediolateral sway of the center of gravity,center of gravity distribution were analyzed before and after the treatment,taking into consideration of visual feedback (eyes open or closed) by using t-test and multiple linear regression analysis.Results Visual feedback had significant no influence on the anteroposterior sway of center of gravity before and after training (P ＞ 0.05) ; Visual feedback had significant influence on mediolateral sway of center of gravity.Rehabilitation training significantly improved the mediolateral sway in opened-eye standing (P ＜ 0.05) but not closedeye standing (P ＞ 0.05).Visual feedback played significant role in center of gravity distribution after training (P ＜ 0.05).Conclusions Visual compensation is of significant influence on the mediolateral sway.Proprioceptive training can help decrease the patient's dependence on visual feedback and enhance the patient's balance ability.

AIM: To test the contents of tanshinone Ⅱ_A and salvianolic acid B in 56 batches of samples by HPLC,evaluate the current quality of Compound Danshen Tables. METHODS: HPLC,chromatographic conditions: The detection column was Agela-C_(18)(5 ?m,46 mm?250 mm) for tanshinone Ⅱ_A,mobile phase was methanol-water(73(∶)27),UV detection wavelength was set at 270 nm.The flow rate was 1 mL/min.The detection column of Agela-C_(18)(5 ?m,4.6 mm?250mm) was used for salvianolic acid with mobile phase of acetonitrile-methanol-formic acid-water(10(∶)30(∶)1(∶)59). UV wavelength was set at 286 nm.The flow rate was 1 mL/min.(RESULTS:) The most of samples conform to the requirement of Chinese Pharmacopoeia(2005),but the content of tanshinone Ⅱ_A was widely different and three batches of salvianolic acid B were lower than the requirement of Chinese Pharmacopoeia(2005). CONCLUSION: The quality of Compound Danshen Tables will be improved.

Aim To isolate cancer stem cells from human pan-creatic cancer cell line L3. 6pl and to identify their biological characteristics. Methods L3. 6pl cells were cultivated in com-mercial low adhesion plate with serum-free stem cell culture me-dium ( MEM/F12 1:1 ) supplemented with B27. The cancer stem cells reformed into floating spheres were isolated. The method of tumor sphere formation was used to isolate/enrich and characterize the cancer stem cells in pancreatic carcinoma cell line L3. 6pl. Cancer stem cell spheres were collected and sorted using magnetic cell sorting ( magnetic activated cell sorting, MACS) technology, with the cell surface markers of CD24 +CD44 + ESA+. Self-renewal and EMT-related oncogene expres-sion were measured with Western blot. Cancer stem cells differ-entiation potential and the expression of cancer stem cell related signs were checked with Immunofluorescence assay. To deter-mine tumorigenesis in vivo, Xenograft assay in NOD-SCID mice were performed respectively, then immunohistochemistry proto-oncogene c-Met and RON expression were also checked. West-ern blot was used to detect the changes of stemness relative tran-scriptional factors and epithelial markers expressed in spheres before and after differentiation. Drug resistance of pancreatic cancer stem cells to gemcitabine or paclitaxel was measured with MTT assay. Results CD24 + CD44 + ESA+ cells were signifi-cantly tumorigenic, and cultured in serum-free conditions to form spheroids, which had the characteristics of stem cells with self-renewal, EMT and drug-resistant capabilities, and had a posi-tive correlation with the c-Met, RON protein expression. Con-clusion Human pancreatic stem cells are successfully isolated, which provides a useful model for individualized therapy and e-valuation of the therapeutic efficacy for pancreatic cancer pa-tients.

OBJECTIVE:To establish the method for the residues determination of 5 organic solvents in teicoplanin raw materi-al and injection. METHODS:Headspace GC was performed on the column with 6% cyanopropylphenyl-94% dimethyl polysiloxane (DB-624)as the stationary phase capillary column,the carrier gas was nitrogen,using the temperature program. The temperature of inlet was 200 ℃,detector was hydrogen flame ionization detector with the flow rate of 1 ml/min,split ratio was 40 ∶ 1 and the vol-ume was 1 ml. RESULTS:Good linearity of ethanol,acetone,ethyl acetate,tetrahydrofuran and triethylamine were obtained(r were 0.999 0-0.999 3);the average recoveries were respectively 95.6%,97.0%,103.2%,94.3%and 98.2%(RSD were 2.1%-4.9%, n=9);RSDs of precision and repeatability tests ≤2.6%;and the minimum detectable concentration were respectively 2,2,2,0.7 and 0.3 μg/ml. CONCLUSIONS:The established method is rapid,sensitive and accurate,and can be used for the residues determi-nation of organic solvents in teicoplanin raw materials and injection.

Objective To investigate the status quo of transformational leadership,psychological empowerment,team psychological safety and team effectiveness,and test whether the supposed structural equation is tenable.Methods Totally 380 nurses completed the Transformational Leadership Questionnaire,Psychological Empowerment Questionnaire,Team Psychological Safety Questionnaire and Team Effectiveness Questionnaire,structural equation model was used to verify the relationship of the above,Bootstrapping confidential interval and product of coefficient were used to test the mediating effect.Results Significant correlations were found between transformational leadership,psychological empowerment,team psychological safety and team effectiveness (P ＜ 0.01),transformational leadership played indirect effect on team effectiveness (Z=6.195,P ＜ 0.05),psychological empowerment and psychological safety had a totally mediation effect.Conclusions Healthcare workers should make psychological empowerment and team psychological safety as the breakthrough point,fully play the advantages of transformational leadership on team effectiveness in order to increase the working ability of nurses and make them complete the clinical work effectively and with high quality.