Objective To compare and analyze the index variation of Yang deficiency mice induced by hydrocortisone in different way of administration and dose. Methods The mice model of Yang deficiency were induced by low dose and high dose hydrocortisone(12.5 mg/kg, 25 mg/kg) in two ways of intramuscular and intraperitoneal injection. The symptoms, body weight, the average intake, the index of stress and the coefficient of sex organs and immune organs of animals were observed. Results In the way of intraperitoneal injection, the weight(24.52±3.29)g, body temperature(35.24±0.32)℃ of high dose of model group were significantly lower than that of control group(31.10±6.11)g,(37.02±0.64)℃. The average intake of low dose group(4.30 ± 0.29)g/(per?d) was lower than control group(7.38±0.53)g/(per?d), and the coefficient of preputial glands(0.10±0.02), penis(0.15±0.03), thymus(0.12±0.03)were lower than that of control group. In the way of intramuscular injection, the average intake of high dose group(5.92±2.01)g/(per?d) was lower than control group(8.60±1.33)g/(per?d). The body temperature(34.90±0.22)℃ and the time of swimming in low temperature (34.00±22.41)s of low dose model group were lower than that of control group(36.43±0.91)℃, (67.17±21.93)s, and the coefficient of thymus of two model groups(0.10±0.02),(0.11±0.06)were lower than that of control group. Conclusion Various dose and model establishment methods of Yang deficiency mice have different the time of symptom appearing, the degree of symptom and sensitive index.

Abstract: To develop novel live attenuated influenza vaccine, we explored the feasibility to attenuate influenza virus by codon deoptimization of NS1. According to the codon usage bias in influenza A virus, we designed and synthesized a condon-deoptimized NS gene by substituting codons of 110 amino acids in the NS1 gene of A/Puerto Rico/8/34(H1N1) with unpreferred synonymous codons. The influenza A virus with the codon deoptimized NS1 gene (deoNS virus) was rescued by reverse genetics. Plaque forming assay and virus growth curve showed that the growth of deoNS virus was reduced about 1000 times in MDCK cells compared to that of the wild-type virus. Intranasal inoculation with deoNS virus did not cause death or evident disease in infected BALB/c mice. Furthermore, the virus titer in the lungs of mice infected with deoNS virus was significantly lower (i.e. 100-1000 times) than that of wild-type virus. Our results indicated that influenza virus could be effectively attenuated by synonymous codon deoptimization of NS1 gene. This strategy will be useful to develop new attenuated candidates for the production of live attenuated influenza vaccines.
Animals ; Base Sequence ; Chick Embryo ; Codon ; genetics ; Influenza A virus ; genetics ; pathogenicity ; Influenza Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Orthomyxoviridae Infections ; immunology ; prevention & control ; Recombination, Genetic ; Vaccines, Attenuated ; immunology ; Viral Nonstructural Proteins ; genetics ; Virulence ; genetics