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Cultivating students' creative spirit was an important aim of higher education,and improving scientific research ability was an effective way to realize this aim.Using Journal of Zhejiang Medical College as the carrier,Journal editorial department tried to improve students' scientific research ability.For instance,served fund projects,found competition's added value,offered course about research design and paper writing.The students' scientific research ability was improved.
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[Objective] To determine the effects against human simplex virus type I(HSV-1) and adenovirus type Ⅲ(ADV-3) anti-viral in vitro and in vivo of eye drop prepared by mixed extract of Heartleaf houttuynia and Lonicera japonica.[Methods] By using different numbers of host cells with cytopathic effect(CPE) as the observation index,models of HSV-1 infected Vero cells and ADV-3 infected HeLa cells were established to determine the anti-viral effects in vitro of HLD with different dosages.Rabbit eye conjunctivitis model infected by HSV-1 was established to observe the anti-viral effects in vivo of HLD with different dosages,and the biopsy specimens of pathogenic eye conjunctive tissue were examined by routine pathological methods.In these experiments,a commercial acilovir eye drop(ACV) was used as the control.[Results] The minimum dosages of HLD to completely inhibit the CPEs of HSV-1 and ADV-3,the host cells were 32 mg/ml and 64 mg/ml respectively.After treatment with HLD,the inflammatory reactions of rabbit eye conjunctivitis caused by HSV-1 infection were obviously weak or disappeared.The curative rates of 1,3 and 6 g/ml HLD treating experimental rabbit eye conjunctivitis were 50.0%,83.3% and 100% respectively.Furthermore,the curative effect using 6 g/ml HLD has no significant difference compared to that using ACV drop.[Conclusion] HLD has a certain anti-viral effect in vitro and a stronger anti-viral effect in vivo,indicating the potential of HLD for developing a new drug to prevent or treat viral eye conjunctivitis.
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AIM: To analyze the nucleotide and putative amino acid sequences of PIA genes isolated from N. gonorrhoeae and to construct the prokaryotic expression system of PIA gene.METHODS: The entire PIA genes from 9 strains of N. gonorrhoeae were amplified by using high fidelity PCR. The target amplification fragments were sequenced after T-A cloning. Homology comparison of the nucleotide and putative amino acid sequences of PIA genes from the isolates with the reported sequences in GenBank was then performed. A prokaryotic expression system of PIA gene was constructed. Different dosages of IPTG were applied to induce the expression of the target recombinant protein (rPIA) and 10% SDS-PAGE plus Bio-Rad Agarose Image Analysor was used to determine the expression level of rPIA. rPIA was extracted using Ni-NTA affinity chromatography and the purified effect was detected by SDS-PAGE.RESULTS: In comparison with the reported PIA gene sequences (GenBank No: L19962), the homologies of nucleotide and putative amino acid sequences of PIA genes from the isolates were 99.6%-100% and 99.1%-100%, respectively, which indicated that all the isolates were belonging to serovars IA6. Output of rPIA was as high as 50.1% of the total bacterial proteins. The purified rPIA only showed a single target protein fragment in gel.CONCLUSION: Serovar IA6 is dominant in the local N. gonorrhoeae isolates and sequences of the encoding gene are relatively conserved. The constructed prokaryotic expression system is able to express rPIA with high efficiency, which may lay a foundation for further development of serological detection kit and vaccine of N. gonorrhoeae.