Objective To explore a new simpler method for the preparation of recombinant alpha virus as a novel vaccine at the DNA level. Methods Plasmids expressing ? gal protein and helper plasmids were transfected into BHK cells. Virus in culture supernatant of the transfected BHK cells were collected and purified and used to infect BHK cells in vitro to identify the expression of target gene and the titre of the recombinant virus. Results Recombinant virus with high titre, prepared by this method, could be expressed well in mammalian cells in vitro . Conclusion High titre recombinant alpha virus can be produced at the DNA level and this method can be applied for vaccine preparation and gene therapy.

At present,there is no unified standards of treatment in elderly advanced non-small cell lung cancer (NSCLC),but to elderly patients with higher KPS scores,radiotherapy or chemotherapy alone has gradually become the main treatment measures.The concurrent chemoradiotherapy can benefit to part of the elderly patients,and it is worth further clinical research.Targeted therapy has exactly effect,and the adverse effect is mild,which provides a new treatment model for elderly advanced NSCLC patients.However,the hospital costs are relatively high,which can not be accepted by ordinary family.So targeted drugs and the model of combination treatment can not be applied extensively.

Objective:To express the GPI-anchored CD55 and recombinant transmembrane form CD55 molecules on mouse fibroblasts cell line NIH3T3, and compare their inhibitory function of complement lysis.Methods:In previous study,had constructed the transmembrane form CD55 (CD55-TM) cDNA by linking the extracellular portion of CD55 to the transmembrane and cytoplasmic domains of MCP, and then subcloned into retroviral vector pLXSN. In this experiment,had transfected recombinant CD55-TM and GPI anchored CD55 into PA317 packaging cell to generate stable virus-producing cell lines. And then, mouse fibroblasts cell line NIH3T3 was infected with the virus containing CD55, recombinant CD55-TM or pLXSN alone. The expression of these molecules on NIH3T3 cells was detected by FACS analysis. Complement killing assay was carried out using MTT colorimetric method.Results:FACS analysis showed that both CD55 and its TM version were expressed stably on NIH3T3 cells. Both kinds of CD55 molecules can efficiently protect the NIH3T3 cells from complement mediated lysis and there is no significant difference between them. Conclusion:These data showed that both GPI-anchored CD55 and its TM version can normally expressed on NIH3T3 cells and both can protect the NIH3T3 cells from human complement mediated lysis. These results confirmed the feasibility of TM CD55 based gene therapy could be used for PNH, and also provided an excellent model for the study of signal transduction mechanisms mediated by GPI-anchored protein.

Objective To construct eukaryotic expression plasmid of hCD137 Fc gene and express hCD137 Fc fusion protein with high biological activity. Methods PCR technique was employed to clone the human CD137 cDNA from a normal human activated T cells cDNA library, and then clone its extramembrane encoding region. The extramembrane sequence together with human IgG1 Fc cDNA were inserted into the eukaryotic expression plasmid pcDNA3. CD137 Fc gene was expressed transiently in 293T cells, and then CD137 Fc protein was purified by recombinated protein A affinity chromatography column. At last, the MW, purity and antigenicity of CD137 Fc were identified by sandwich ELISA, SDS PAGE and Western blotting, respectively. Results The ORF of CD137 Fc gene was coincident with what we expected. ELISA and SDS PAGE confirmed protein expression in 293T cells. Western blotting proved the antigenicity of the purified CD137 Fc protein. Conclusion We obtained a purified recombinated CD137 Fc protein with biological activity and the expected MW. This lays the foundation for further studies of CD137 such as its role in immune homeostasis and other biological functions.

Objective To clone the full length cDNA of human LIGHT and to construct the recombinant eukaryotic expression plasmid pCI neo LIGHT for the stable expression on 293T cells. Methods Human LIGHT cDNA was cloned from a normal human activated T cell cDNA library phAD.CAD by PCR. After sequencing, LIGHT cDNA was inserted into plasmid pCI neo for the construction of the eukaryotic expression. The recombinant was transfected into 293T cells by electroporation. The expression of LIGHT on the surface of 293T cells was detected by flow cytometry after screening with G418. Results Sequencing confirmed that ORF of LIGHT gene was intact and right. Restrictive enzyme digestion proved that LIGHT gene was inserted into the recombinant plasmid of LIGHT pCI neo correctly. FACS analysis revealed that about 78.69% 293T cells expressed LIGHT protein on the cell surfaces at 3 months after screening with G418. Conclusion LIGHT gene has been cloned successfully, and a 293T cell line expressing LIGHT protein on its membrane surface has been obtained.

Objective To explore the clinical efficacy of the interventional recanalization for infertile women with fallopian tube obstruction. Methods 636 infertile patients with fallopian tube obstruction diagnosed by hystero-salpin-gngraphy were treated with interventional tubal recanalizafion under X-ray. Results Among 636 patients with 984 complete fallopian tubes obstruction,there were 804 recanalization in tile 984 fallopian tubes and the successful rate of recanalization was 81.71%. Among 252 fallopian tubes with not complete obstruction ,there were 252 recanali-zation in the 252 fallopian tubes and the successful rate of recanalization was 100%. The all successful rate of reca-nalization was 88.3%. These patients pregnancy rate was 28.30% in one year and the second fallopian tube obstruc-tion rate was 6.79%. There was 4 ectopic pregnancy. Conclusions Interventional tubal recanalization has important application in the diagnosis and treatment for infertile women with fallopian tube obstruction, h has merit, such as simple, safe and good effect.

Objective To investigate the effects of training of Bobath position transfer technique for nurses.Methods Thirty nurses were recruited in the study using convenience sampling and received training of Bobath transfer technique.Five variables were evaluated before and after training:transfer skills,transfer intensity,sense of comfort,and sense of comfort and sense of security from simulated patients.Results Nurses' transfer skills and sense of comfort were higher after training (P＜0.05).Transfer intensity was lower than before (P＜0.05).Sense of comfort and sense of security from simulated patients were improved after the training (P＜0.05).Conclusion Training of Bobath transfer technique can improve nurses' transfer skills and promote sense of comfort and sense of security from simulated patients.

Objective To evaluate the effects of metformin on thyrotropin(TSH)levels. Methods From the patients with type 2 diabetes mellitus or metabolic syndrome, 48 patients with primary hypothyroidism were enrolled and grouped. 17 patients were treated only with metformin(group A), 19 patients with metformin and stable L-T4substitution(group B), and the remaining 12 patients with antidiabetic drugs(other than metformin)and L-T4(group C). Meanwhile, 20 euthyroid patients with other thyroid abnormalities(group D)and 30 patients without thyroid diseases(group E)served as control. TSH, FT3, FT4, TT3, TT4, and blood glucose were determined regularly in all these subjects. Results After administration of metformin for 12 months, serum TSH were decreased in group A [(5.05±1.07 vs 2.61±0.91)mU/L, P＜0.01] and group B [(2.67±1.03 vs 1.35±0.74)mU/L, P＜0.01]. No difference was found in FT3and FT4in both groups. TSH levels were raised from(1.30±0.71)to(2.58±1.02)mU/L(P＜0.01)within 8～12 weeks in 13 out of 15 patients after metformin withdrawal. Serum TSH and thyroid hormones in the other 3 groups were not significantly changed. Conclusion Administration of metformin may lead to reduction of serum TSH level.