Study on foam sclerotherapy for varicose veins of lower extremities / 临床外科杂志

Objective To explore the relationship between recurrence rate and side effects of the treatment of varicose veins in lower extremity with lauromacrogol foam sclerosing agent.Methods The data of 62 patients(98 limbs)with varicose veins was collected,including 27 males (41 limbs)and 35 females (57 limbs).They were treated with 1％ lauromacrogol foam sclerosing agent combined with great saphenous vein high ligation and endovenous laser treatment.Statistical indicators included side effects such as pain,induration,hyperpigmentation,swelling,numbness and the recurrence rate.Results Two patients were lost,57 patients were injected once,further foam sclerotherapy was carried out again for 3 patients.Obvious abnormal varicose veins as well as the soreness and fatigue of lower extremity disappeared in all patients.Thirty-seven patients had pain and 43 patients developed superficial venous thrombotic sclerosis,among which 50 patients faded away after 1 to 3 months by taking diosmin.Thirty-one cases of pigmentation occurred and 20 disappeared after 1 to 3 months by applying the vitamin E whitening essence.Six patients with lower extremity ulcers recovered after 1 to 3 months.Through statistics analysis of the patients in 1-month follow-up,we found that the differences in recurrence rate after 6 months were statistically significant between group with pain induration reaction and group without (P ＜ 0.05).Among the patients without using the vitamin E whitening essence,there was a significant correlation between the duration of pigmentation and recurrence rate after 6 months.Conclusion In the process of foam sclerotherapy,the more pain induration reaction they had and the longer the duration of pigmentation sustained,the lower the recurrence rate would be accordingly.

Isolation of specific humanized anti-D-dimer scFv fragments from scFv phage libraries / 中华微生物学和免疫学杂志

Objective To isolate specific humanized anti-D-dimer scFv(single chain Fv) antibody from scFv phage libraries. Methods Isolate anti-D-dimer positive clones from Tomlinson I + J phage libraries by three rounds of panuing, then sequence monoclonal genes by bideoxy-mediated chain termination and express soluble scFv antibody; Pick out anti-D-dimer antibodies with high specificity and affinity by ELISA.Results After three rounds of selection from human scFv phage libraries Tomlinson I and J, 38 monclonal specific anti-D-dimer scFv fragments were selected. By polyclonal and monoclonal phage ELISA and gene sequencing, 20 different full-length monoclonal scFv phages were identified, the result of soluble scFv ELISA showed that 20 full-length monoclonal scFv were expressed smoothly. According to the result of soluble scFv ELISA, in 5 scFv antibodies with high value of A450 selected, 3 scFv antibody fragments showed high specific and affinity. Conclusion Antibody phage display was an effective, rapid method to isolate anti-D-dimer antibodies with high specificity and affinity.

Construction and expression of anti-CD62P/anti-GPIIb-Illa diabody

To construct 3 expression plasmids for the targeted therapy of thrombosis: single chain variable fragment [scFv] of monoclonal antibody [mAb] 7E3 that can identify and bind platelet glycoprotein GPIIb-IIIa complex, scFv of mAb WAPS12.2 that can identify and bind P selectin [CD62P], a diabody that can identify and bind GPIIb-IIIa and CD62P imultaneously, and to investigate whether the vectors can express correctly. This study was carried out at the Laboratory of Hunan Yuantai Biological Technology Co. Ltd, Hubei, China from September 2007 to May 2008. Total RNA of mAb 7E3 cells and WAPS12.2 cells were obtained. Reverse transcriptase polymerase chain reaction [PCR] was carried out to obtain the genes of variable regions of light and heavy chains of 7E3 and WAPS12.2. Target genes were named 7E3VL, 7E3VH, CD62PVL, and CD62PVH. The 7E3VL-7E3VH and CD62PVL-CD62PVH were obtained by PCR and connected with pET-22b[+]. Products were named pET-scFv7E3 and pET-scFvCD62P. The 7E3VL-CD62PVH and CD62PVL-7E3VH were obtained by using overlap PCR and were ligated to pET-22b[+]. The products were named pET-ED1 and pET-ED2. The PCR was performed by taking pET-ED2 as a template to obtain the complete operon gene and was ligated to pET-ED1. The product was named pET-7ECD. The identification by restriction endonuclease cleavage and DNA sequencing confirmed that the construction of these expression plasmids was successful. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot confirmed that these plasmids expressed correctly. The expression plasmids pET-scFv7E3, pET-scFvCD62P, and pET-7ECD were constructed and expressed successfully, and laid a good foundation for further research on target-oriented thrombolytic agents

Construction of adeno-associated virus coexpression system for human angiopoietin-1 and VEGF gene / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Ischemic disease is one of the leading causes of death in the world. In order to further study gene therapy for ischemic disease, we constructed a recombinant plasmid for co-expression of human angiopoietin-1 and vascular endothelial growth factor 165(VEGF165) gene in adeno-associated virus (AAV) gene delivery system.</p><p><b>METHODS</b>Human angiopoietin 1 and VEGF165 gene were obtained using PCR. The upstream of angiopoietin 1 contained restriction enzyme site HindIII, and the downstream of angiopoietin 1 contained restriction enzyme site BamHI. The upstream of VEGF165 contained restriction enzyme site BglII, and the downstream of VEGF165 contained restriction enzyme site BamHI. Using the multiple cloning sites (MCS) in plasmid pZero++ such as BamHI, BglII, HindIII, NotI, XhoI, XbaI, SalI, BspHI, KspI and the corresponding MCS in plasmid pAAV-MCS, angiopoietin 1 and VEGF165 gene were subcloned into pAAV-MCS.</p><p><b>RESULTS</b>DNA sequencing revealed that the PCR- amplified angiopoietin 1 and VEGF165 were consistent with NCBI Gene Bank. The recombinant plasmid was identified using PCR and digestion, which proved to be consistent with our hypothesis. In recombinant plasmid, angiopoietin1 and VEGF possessed a CMV promoter and polyA terminator system respectively, thus assuring co-expression of the two genes.</p><p><b>CONCLUSION</b>Successful construction of AAV co-expression system for human angiopoietin 1 and VEGF165 gene will provide the foundation for gene therapy to cure severe ischemic disease.</p>