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Objective:To express virus-like particles of poliovirus type 2 (PV2-VLP) in insect cells using a recombinant baculovirus expressing P1 and 3CD and to preliminarily evaluate its immunogenicity.Methods:Based on the codon preference of High 5 cells, the sequences of P1 gene and 3CD gene of PV2 were optimized and inserted into pUC57-Amp to construct pUC57-PV2-P1 and pUC57-PV2-3CD. UC57-PV2-P1s mutant that carried P1 gene mutation affecting thermostability was then constructed. Recombinant baculovirus strains of rBac-PV2-P1s-3CD and rBac-PV2-P1-3CD (wild type) were constructed using homologous recombination. The expression of target proteins was detected by Western blot. PV2-VLP was purified by ion exchange chromatography. The structure of VLP was observed under transmission electron microscopy to evaluate the assembly efficiency. The immunogenicity of PV2-VLP was assessed in a rat model.Results:The recombinant baculovirus with stable expression of P1s and 3CD proteins was successfully constructed. Western blot results showed that the yield of VLP was higher after thermostability mutation than that of the wild type. A three-dimensional structure with a diameter of about 30 nm was observed under electron microscopy, indicating that the VLP was successfully assembled. Animal experiment showed that the recombinant PV2-VLP had immunogenicity and could effectively induce the production of neutralizing antibodies.Conclusions:Effective VLP vaccines could be successfully prepared using the insect cell-baculovirus expression system, which provided reference for the development of polio VLP vaccine.
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Objective Conducting phacoemulsification combined with trabeculectomy through double incision to treat glaucoma and cat-aract,and observing the visual acuity,intraocular pressure before and after the operation and complications of this therapy. Methods Totally 60 patients (78 eyes) with glaucoma and cataract from January 2010 to December 2013 in our hospital were observed. 32 patients (40 eyes) who were given phacoemulsification combined with trabeculectomy through double incision were regarded as the observation group,and the other 28 patients (38 eyes) who were merely given phacoemulsification were regarded as the control group. The visual acuity,intraocular pres-sure and anterior chamber of the two groups were observed and recorded,and they were followed up for 36 months. Results Visual acuity and intraocular pressure of the two groups were improved after operation,but the observation group achieved a better result which showed a significant difference (P< 0. 05). Anterior chamber inflammation occurred in patients of the observation group,and 2 cases were of anterior chamber exudation. The control group,by contrast,has less anterior chamber inflammation. There was no hyphema,shallow anterior chamber, choroidal detachment occurred in both of the two groups. Conclusion Using phacoemulsification combined with trabeculectomy through double incision to treat glaucoma and cataract could receive a good recovery of visual acuity and intraocular pressure. Although,more severe anterior chamber inflammation occurred after operation compared with the control group,it can be alleviated soon after medication. This meth-od is a good therapy which can reduce pain and economic burden of patients from multiple operations. But it requires a strict selection of ap-propriate case.