AIM To test whether Nox1 plays a role in typhlitis induced by Salmonella enterica serovar Typhimurium(S. Tm) in a mouse model.METHODS Eight-week-old male wild-type(WT) and Nox1 knockout(KO) C57BL6/J(B6) mice were admi...AIM To test whether Nox1 plays a role in typhlitis induced by Salmonella enterica serovar Typhimurium(S. Tm) in a mouse model.METHODS Eight-week-old male wild-type(WT) and Nox1 knockout(KO) C57BL6/J(B6) mice were administered metronidazole water for 4 d to make them susceptible to S. Tm infection by the oral route. The mice were given plain water and administered with 4 different doses of S. Tm by oral gavage. The mice were followed for another 4 d. From the time of the metronidazole application, the mice were observed twice daily and weighed daily. The ileum, cecum and colon were removed for sampling at the fourth day post-inoculation. Portions of all three tissues were fixed for histology and placed in RNAlater for m RNA/c DNA preparation and quantitative real-time PCR. The contents of the cecum were recovered for estimation of S. Tm CFU.RESULTS We found Nox1-knockout(Nox1-KO) mice were not more sensitive to S. Tm colonization and infection than WT B6 mice. This conclusion is based on the following observations:(1) S. Tm-infection induced similar weight loss in Nox1-KO mice compared to WT mice;(2) the same S. Tm CFU was recovered from the cecal content of Nox1-KO and WT mice regardless of the inoculation dose, except the lowest inoculation dose(2 × 106 CFU) for which the Nox1-KO had one-log lower CFU than WT mice;(3) there is no difference in cecal pathology between WT and Nox1-KO groups; and(4) there are no S. Tm infection-induced changes in gene expression levels(IL-1b, TNF-α, and Duox2) between WT and Nox1-KO groups. The Alpi gene expression was more suppressed by S. Tm treatment in WT than the Nox1-KO cecum. CONCLUSION Nox1 does not protect mice from S. Tm colonization. Nox1-KO provides a very minor protective effect against S. Tm infection. Using NOX1-specific inhibitors for colitis therapy should not increase risks in bacterial infection.展开更多
To investigate the anti-vasculature effects and the anti-glioma effects of attenuated Salmonella typhimurium vaccine strain expressing VEGFR2 (flk-1) gene, plasmid pcDNA3.1-flk1 was constructed and electro-transfected...To investigate the anti-vasculature effects and the anti-glioma effects of attenuated Salmonella typhimurium vaccine strain expressing VEGFR2 (flk-1) gene, plasmid pcDNA3.1-flk1 was constructed and electro-transfected into live attenuated Salmonella typhimurium strain SL7207. Mouse models of intracranial Gl261 glioblastoma were treated with an orally administered attenuated Salmonella typhimurium expressing flk-1 gene. The survival period was recorded and vessel density was observed by immunofluorescence. CTLs activity was measured by MTT assay. Our results showed that attenuated Salmonella typhimurium vaccine strain expressing flk-1 gene could significantly inhibit glioblastoma growth, reduce vessel density, prolong the survival period and improve the survival rate in these mice. The flk-1 specific CTLs activity was increased obviously after the vaccination. Our study showed that attenuated Salmonella typhimurium vaccine strain expressing flk-1 gene could break peripheral immune tolerance a in glioma gainst this self-antigen and kill endothelial cells by the orally administered vaccine and can be used for both prophylactic and therapeutic purposes.展开更多
目的1株mphA和mcr-1阳性鼠伤寒沙门菌的鉴定及耐药特征分析;方法分离自4岁腹泻患者粪便样本中的1株鼠伤寒沙门菌,使用沙门菌血清凝集法进行菌株鉴定,质谱法复核鉴定结果;使用微量肉汤稀释法进行药敏试验;通过全基因组测序及耐药基因注...目的1株mphA和mcr-1阳性鼠伤寒沙门菌的鉴定及耐药特征分析;方法分离自4岁腹泻患者粪便样本中的1株鼠伤寒沙门菌,使用沙门菌血清凝集法进行菌株鉴定,质谱法复核鉴定结果;使用微量肉汤稀释法进行药敏试验;通过全基因组测序及耐药基因注释鉴定菌株所携带的耐药基因;通过质粒测序及生信分析进行质粒类型鉴定、质粒耐药基因、插入子等注释及质粒结构分析;通过质粒接合转移试验评估质粒的水平转移能力。结果药敏试验结果显示分离株对包括阿奇霉素、多黏菌素、头孢曲松和环丙沙星等临床常用抗生素药物耐药。耐药基因鉴定结果显示:该分离株携带包括大环内酯类抗性基因mphA,多黏菌素抗性基因mcr-1在内的12类45个耐药基因及1个氟喹诺酮类耐药点突变gyr A S83F。质粒测序分析结果显示:该菌株携带一个260,008bp大小的Inc HI2型质粒,该质粒携带包括mphA基因和mcr-1基因在内的19个耐药基因,15种插入子,4种转座子。质粒接合转移实验结果显示该质粒可以在肠杆菌目细菌之间水平转移。结论研究结果为临床治疗鼠伤寒沙门菌感染中合理使用抗生素以及有效预防、控制耐阿奇霉素、多黏菌素鼠伤寒沙门菌的传播提供了科学依据。展开更多
supported by the Earmarked Fund for the Modern Agroindustry Technology Research System, China （CARS-41）;the National High Technology Research and Development Program of China （2011AA100301）
基金Supported by Federal funds from the National Cancer Institute(NCI)under Contract,No.HHSN261200800001E(to Chu FF)Research reported in this publication included work performed in the Animal Resources Center Core supported by the National Cancer Institute of the National Institutes of Health under award No.P30CA033572
文摘AIM To test whether Nox1 plays a role in typhlitis induced by Salmonella enterica serovar Typhimurium(S. Tm) in a mouse model.METHODS Eight-week-old male wild-type(WT) and Nox1 knockout(KO) C57BL6/J(B6) mice were administered metronidazole water for 4 d to make them susceptible to S. Tm infection by the oral route. The mice were given plain water and administered with 4 different doses of S. Tm by oral gavage. The mice were followed for another 4 d. From the time of the metronidazole application, the mice were observed twice daily and weighed daily. The ileum, cecum and colon were removed for sampling at the fourth day post-inoculation. Portions of all three tissues were fixed for histology and placed in RNAlater for m RNA/c DNA preparation and quantitative real-time PCR. The contents of the cecum were recovered for estimation of S. Tm CFU.RESULTS We found Nox1-knockout(Nox1-KO) mice were not more sensitive to S. Tm colonization and infection than WT B6 mice. This conclusion is based on the following observations:(1) S. Tm-infection induced similar weight loss in Nox1-KO mice compared to WT mice;(2) the same S. Tm CFU was recovered from the cecal content of Nox1-KO and WT mice regardless of the inoculation dose, except the lowest inoculation dose(2 × 106 CFU) for which the Nox1-KO had one-log lower CFU than WT mice;(3) there is no difference in cecal pathology between WT and Nox1-KO groups; and(4) there are no S. Tm infection-induced changes in gene expression levels(IL-1b, TNF-α, and Duox2) between WT and Nox1-KO groups. The Alpi gene expression was more suppressed by S. Tm treatment in WT than the Nox1-KO cecum. CONCLUSION Nox1 does not protect mice from S. Tm colonization. Nox1-KO provides a very minor protective effect against S. Tm infection. Using NOX1-specific inhibitors for colitis therapy should not increase risks in bacterial infection.
文摘To investigate the anti-vasculature effects and the anti-glioma effects of attenuated Salmonella typhimurium vaccine strain expressing VEGFR2 (flk-1) gene, plasmid pcDNA3.1-flk1 was constructed and electro-transfected into live attenuated Salmonella typhimurium strain SL7207. Mouse models of intracranial Gl261 glioblastoma were treated with an orally administered attenuated Salmonella typhimurium expressing flk-1 gene. The survival period was recorded and vessel density was observed by immunofluorescence. CTLs activity was measured by MTT assay. Our results showed that attenuated Salmonella typhimurium vaccine strain expressing flk-1 gene could significantly inhibit glioblastoma growth, reduce vessel density, prolong the survival period and improve the survival rate in these mice. The flk-1 specific CTLs activity was increased obviously after the vaccination. Our study showed that attenuated Salmonella typhimurium vaccine strain expressing flk-1 gene could break peripheral immune tolerance a in glioma gainst this self-antigen and kill endothelial cells by the orally administered vaccine and can be used for both prophylactic and therapeutic purposes.
文摘目的1株mphA和mcr-1阳性鼠伤寒沙门菌的鉴定及耐药特征分析;方法分离自4岁腹泻患者粪便样本中的1株鼠伤寒沙门菌,使用沙门菌血清凝集法进行菌株鉴定,质谱法复核鉴定结果;使用微量肉汤稀释法进行药敏试验;通过全基因组测序及耐药基因注释鉴定菌株所携带的耐药基因;通过质粒测序及生信分析进行质粒类型鉴定、质粒耐药基因、插入子等注释及质粒结构分析;通过质粒接合转移试验评估质粒的水平转移能力。结果药敏试验结果显示分离株对包括阿奇霉素、多黏菌素、头孢曲松和环丙沙星等临床常用抗生素药物耐药。耐药基因鉴定结果显示:该分离株携带包括大环内酯类抗性基因mphA,多黏菌素抗性基因mcr-1在内的12类45个耐药基因及1个氟喹诺酮类耐药点突变gyr A S83F。质粒测序分析结果显示:该菌株携带一个260,008bp大小的Inc HI2型质粒,该质粒携带包括mphA基因和mcr-1基因在内的19个耐药基因,15种插入子,4种转座子。质粒接合转移实验结果显示该质粒可以在肠杆菌目细菌之间水平转移。结论研究结果为临床治疗鼠伤寒沙门菌感染中合理使用抗生素以及有效预防、控制耐阿奇霉素、多黏菌素鼠伤寒沙门菌的传播提供了科学依据。
基金supported by the Earmarked Fund for the Modern Agroindustry Technology Research System, China (CARS-41)the National High Technology Research and Development Program of China (2011AA100301)
文摘supported by the Earmarked Fund for the Modern Agroindustry Technology Research System, China （CARS-41）;the National High Technology Research and Development Program of China （2011AA100301）
