Objective To investigate and analyze the health education requirement in migrant-worker patients and establish health education countermeasures that accord with migrant-worker pa-tients. Methods 600 hospitalization patients were chosen from January to December, 2007,among which 300 migrant-worker patients were set as the experimental group,300 native patients were selected as the control group.The contents and manners of health education requirement were compared between the two groups. Results In the aspect of contents of health education requirement,the highest require-ment was to understand treatment expense,which was significantly higher than that of the control group. The higher requirement included to understand illness condition and treatment protocols,prevention and treatment of common disease and infectious disease.In the aspect of manners of health education require-ment,the highest requirement was to communicate with medical workers,the lowest requirement was to communicate among patients. Conclusions Migrant-worker patients concern about treatment expense, nurses should not only conduct health education about disease-related knowledge, but also pay much atten-tion to informed consent about treatment expense in order to avoid unnecessary dispute.

Objective To extract hyperoside from the leaves of Acanthopanax sessiliflorus by complex enzyme method, and optimize the extraction process by orthogonal experiment. Methods Hyperoside was determined by HPLC. Effects of temperature,α-amylase, neutral protease and cellulase on extraction rate were detected by the orthogonal tests, and the optimum extraction condition of hyperoside from the leaves of Acanthopanax sessiliflorus was determined by complex enzyme method. Results The main influence factor was temperature,follows byα-amylase, neutral protease and cellulase according to orthogonal analysis.The best condition was as follows: dose of cellulase, neutral protease and α-amylase was 2%, 0. 5% and 3%, respectively, extract at temperature of 30 ℃for 10 min. Under this condition, the extraction rate of hyperoside in the leaves of Acanthopanax senticosus was 0.52%. Conclusion As compared with the traditional technics, compound enzyme increases the productivity of hyperoside.

This study was aimed to optimize the extraction process of hyperoside from leaves ofAcanthopanax senticosusby compounding-enzyme method and orthogonal experiment. The hyperoside compound was regarded as standard and determined by HPLC. Based on the experiments of 4 factors including the enzyme amount, temperature, extraction time and PH values, the extraction process of hyperoside was determined by the orthogonal experiments and variance analysis. The results of single-factor experiments showed that different enzymes showed different effects on the enhance yield of hyperoside. The effects of different factors showed that the order of PH, neutral protease, temperature, time, pectinase, xylanase and cellulose was from strong to weak. Through orthogonal analysis, the optimum conditions were 2% pectinase, 2% xylanase, 0.5% neutral protease, and 0.5% cellulose, under the temperature of 30°C, extraction time of 10 min, and pH = 4.5. Under these conditions, the extraction rate was 1.84%. The yield was increased 107% compared with traditional process. It was concluded that the use of compounding enzyme can increase the yield of hyperoside, which possessed a lot of economic benefits.

Objective:To prepare a novel tri-specific T cell engager (19TriTE) targeting CD19 antigen, and to investigate its immunotherapeutic effect on CD19-positive hematological malignancies.Methods:19TriTE was constructed by molecular cloning technology and successfully expressed through the eukaryotic expressing system. The effects of 19TriTE on the proliferation and activation of T cells, as well as the specific cytotoxicity against CD19 positive tumor cell lines were verified.Results:①19TriTE expressing plasmid was constructed and successfully expressed through the eukaryotic expressing system. ②19TriTE can specifically bind to T cells and Nalm6 cells, with equilibrium dissociation constants of 19.21 nmol/L and 11.67 nmol/L, respectively. ③The expression rates of CD69 positive T cells and CD25 positive T cells were 35.4% and 49.8% respectively, when 2 nmol/L 19TriTE were added in the co-culture system, which were significantly higher than those in the control group. ④19TriTE can significantly promote the proliferation of T cells. The absolute count of T cells expanded from the initial one million to 74 million with an 74 fold increase at the concentration of 1 nmol/L on day 12. ⑤19TriTE can significantly mediate T cells killing of CD19 positive target cells in a dose-dependent manner. At the concentration of 10 nmol/L, the target cells lysis reached 50%. ⑥Degranulation experiment verified that 19TriTE can activate T cells in the presence of CD19 positive target cells, and the activation of T cells positively correlated with the dose of 19TriTE. ⑦When 19TriTE fusion protein co-cultured with T cells and target cells overexpression RFP and luciferase genes respectively, 19TriTE can notably mediate T cells killing of CD19 positive target cells through fluorescent microscope or bioluminescence imaging technology.Conclusion:In this study, we successfully constructed and expressed 19TriTE fusion protein and verified that it can effectively activate T cells and promote their proliferation in vitro. At the same time, it can bind to CD19 positive target cells and T cells, as well as enhance T cells anti-leukemia effect in vitro, providing the foundation for further clinical research.

Objective:To construct a novel dual-specific antibody targeting human CD123 (CD123 DuAb) and study its effects in acute myeloid leukemia (AML) .Methods:Based on the variable region of the CD123 monoclonal antibody independently developed at our institution, the CD123 DuAb expression plasmid was constructed by molecular cloning and transfected into ExpiCHO-S cells to prepare the antibody protein. Through a series of in vitro experiments, its activation and proliferation effect on T cells, as well as the effect of promoting T-cell killing of AML cells, were verified.Results:① A novel CD123 DuAb plasmid targeting CD123 was successfully constructed and expressed in the Expi-CHO eukaryotic system. ②The CD123 DuAb could bind both CD3 on T cells and CD123 on CD123 + tumor cells. ③When T cells were co-cultured with MV4-11 cells with addition of the CD123 DuAb at a concentration of 1 nmol/L, the positive expression rates of CD69 and CD25 on T cells were 68.0% and 44.3%, respectively, which were significantly higher than those of the control group ( P<0.05). ④Co-culture with CD123 DuAb at 1 nmol/L promoted T-cell proliferation, and the absolute T-cell count increased from 5×10 5/ml to 3.2×10 6/ml on day 9, and CFSE fluorescence intensity decreased significantly. ⑤ With the increase in CD123 DuAb concentration in the culture system, T-cell exhaustion and apoptosis increased. When the CD123 DuAb was added at a concentration of 1 nmol/L to the culture system, the proportion of CD8 + PD-1 + LAG-3 + T cells was 10.90%, and the proportion of propidium iodide (PI) - Annexin Ⅴ + T cells and PI + Annexin Ⅴ + T cells was 18.27% and 11.43%, respectively, which were significantly higher than those in the control group ( P<0.05). ⑥ The CD123 DuAb significantly activated T cells, and the activation intensity was positively correlated with its concentration. The expression rate of CD107a on T cells reached 16.05% with 1 nmol/L CD123 DuAb, which was significantly higher than that of the control group ( P<0.05). ⑦The CD123 DuAb promoted cytokine secretion by T cells at a concentration of 1 nmol/L, and the concentration of IFN-γ and TNF-α in the supernatant of the co-culture system reached 193.8 pg/ml and 169.8 pg/ml, respectively, which was significantly higher than that of the control group ( P<0.05). ⑧When CD123 DuAb was added at a concentration of 1 nmol/L to the co-culture system of T cells and CD123 + tumor cells, the killing intensity of T cells significantly increased, and the residual rates of CD123 + MV4-11 cells, CD123 + Molm13 cells, and CD123 + THP-1 cells were 7.4%, 6.7%, and 14.6% on day 3, respectively, which were significantly lower than those in the control group ( P<0.05) . Conclusion:In this study, a novel CD123 DuAb was constructed and expressed. In vitro experiments verified that the DuAb binds to CD123 + tumor cells and T cells simultaneously, promotes T-cell activation and proliferation, and facilitates their anti-leukemia effect, which provides a basis for further clinical research.