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1.
International Journal of Pediatrics ; (6): 669-672, 2015.
Artigo em Chinês | WPRIM | ID: wpr-483243

RESUMO

With the development and wide application of antibiotics, the number of resistant strains keeps growing and there is a trend towards a higher isolation rate of methicillin-resistant staphylococcus aureus (MRSA) year by year, accompanied by the increasing drug resistance.As reported in recent years, vancomycin resistant or intermediate staphylococcus aureus was isolated from different geographical regions.The prevalence of MRSA infection has become a serious public health problem, which is also the global focus in the medical field.This review focuses on the prevalence of MRSA colonization and infection, drug resistance mechanism,mode of transmission, prevention and efficacy in neonatal intensive care unit.

2.
Chinese Journal of Microbiology and Immunology ; (12): 604-609, 2013.
Artigo em Chinês | WPRIM | ID: wpr-437304

RESUMO

Objective To express virus-like particles(VLP) of enterovirus 71 (EV71) in Han-senula polymorpha.Methods The coding sequences of P1 and 3CD genes of EV71 were optimized accord-ing to codon usage bias of Hansenula polymorpha for achieving high expression , and then cloned into the ex-pression vector PMV of Hansenula polymorpha .The recombinant expression vector PMV-P1-3CD was trans-formed into Hansenula polymorpha AU 0501 .The transformants were stably cultured in selective medium (Yeast Nitrogen Base) and screened for strains with positive P1 and 3CD genes by PCR.Then an induced cultivation on the recombinant strains were performed in a medium supplemented with methanol to a final concentration of 1.0%and the expressed products were analyzed by SDS-PAGE and Western blot assays to select high expression strains .The high expression strains were cultured in 30 L fermentor and its fermenta-tion products were analyzed by electronic microscope after purification .Results EV71 recombinant expres-sion strains were successfully constructed .The results of SDS-PAGE showed that the expressed products had obvious VP3, VP1, VP0 protein bands with molecular weights of 26×103, 33×103 and 35×103, respective-ly, which were consistent with the expected molecular weight of the fusion proteins .Western blot demonstra-ted that the expressed products could be specifically recognized by the polyclonal antibody against EV 71-VP1 at 33 ×10 3 , indicating its high immunoreactivity .ELISA confirmed that the expression level of EV 71 fermen-tation products was reached to 200 mg/L.Electronic microscope analysis showed that the VLP of recombi-nant EV71 were 24-30 nm in diameter with normal structure .Conclusion The virus-like particles of human enterovirus 71 are successfully expressed in Hansenula polymorpha , which provides a foundation for the fur-ther development of EV 71 VLP vaccine .

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