Objective To establish a method for isolating alveolar macrophage (AM) of mouse based on flow cy tometry.Methods The lungs were digested by collagen ⅣV in vitro to prepare single-cell suspension that was stained by CD11 b and CD1 1 c antibody.CD1 1 b1owCD1 1 c + cell population were AM and isolated by flow cytometry.After that,the cell viability was measured via the Typan blue staining,and the identification of AM was through flow cytometry and real-time PCR.Results CD1 1 b1owCD1 1 c + cell population was isolated by flow cytometry,the purity was (93 ± 2)％ and the cell viability was (80 ±5)％.The real-time PCR results showed that peroxisome proliferator-activated receptor γ (PPARγ) mRNA was highly expressed in AM isolated by flow cytometry (P ＜ 0.001).In addition,the functional assay showed that the isolated AM possess high phagocytic activity.Thus,the results described above demonstrate that the isolated cells were AM.Conclusion A method for obtaining AM based on flow cytometry was established.The method has high cell purity and good cell activity which can be used for functional experiments.

OBJECTIVE:To describe the characteristics of existing numerical methods,conclude current mainstream bio-electromagnetism simulation softwares,focusing on their different directions,and explore the developing trends of bio-electromagnetism computing platform.DATA SOURCES:A computer-based online search of Wanfang,Sciencadirect (Elsevier),and IEEE database was performed for articles published between 1999 and 2009 with the key words "bio-electromagnetism,simulation,numerical simulation,FDTD* in Chinese and English.A total of 83 articles were collected,including 28 Chinese and 55 English.Four software introduction and user manuals were manually retrieved.DATA SELECTION:Articles were screened by reading the title and abstract,and the latest relevant research literatures in the same kinds of simulation methods were included.MAIN OUTCOME MEASURES:A total of 25 articles with no related content,and 27 repetitive and outdated studies were excluded.Finally,31 Chinese and English articles were included for further analysis,involving 8 reviews and comments and 23 original articles.RESULTS:Common electromagnetic field numerical calculations include time domain finite difference method,moment method,and finite element method.Currently,the bio-electromagnetism simulation softwares used in electromagnetic radiation on organism include SEMCAD-X,FEKO,XFDTD,HFSS and GEMS.CONCLUSION:Future bio-electromagnetic computing platform is expected to integrate a variety of numerical methods to meet the research needs of a variety of simulation objects.In addition,interface facilitation of simulation and flowsheet of modeling will be the focus of future improvements.

Objective : Tnfrsf11 a Cre Rosa26 yfp reporter gene mice were prepared to determine the efficiency of Cre-mediated recombination using flow cytometry in different tissue-resident macrophages. Methods : TheTnfrsf11 a Cre Rosa26 yfp reporter mice were generated by crossingTnfrsf11 a Cre mice withRosa26 yfp mice and identified by PCR.Brain microglia, liver macrophages, kidney macrophages, alveolar macrophages and spleen macrophages were separated from adultTnfrsf11 a Cre Rosa26 yfp reporter mouse and yellow fluorescent protein(YFP) labeling efficiency was analyzed by flow cytometry. Results : YFP expression percentage was about 91.27% of brain microglia inTnfrsf11 a Cre Rosa26 yfp reporter mice, but liver, spleen, and alveolar macrophages were inefficiently targeted(63.60%,69.66%,32.76%). Conclusion Tnfrsf11 a Cre mice were qualified as conditional knockout model mice of brain microglia andTnfrsf11 a Cre Rosa26 yfp reporter mice can be used as a tool for conditional knockout of brain microgliain vivo.

Objective : To trace and investigate the distribution of hepatic stellate cells by Cre⁃Loxp technology of LratCre mice. Methods : LratCre mice were mated with H11LoxP⁃ZsGreen⁃Stop⁃LoxP⁃tdTomato reporter mice and genotype of their progeny was identified by PCR. The liver sections were fixed , the antibody of liver non⁃parenchymal cells was labeled with immunofluorescence , and the expression and distribution of liver stellate cells and other liver non⁃parenchymal cells were analyzed by Imaris 3D reduction , as well as positively identify dual positioning of hepatic stellate cells. Results : LratCre H11LoxP⁃ZsGreen⁃Stop⁃LoxP⁃tdTomato reporter mice specificiallyled to expression of red fluorescent protein tdTomato in hepatic stellate cells , providing a way to analyze the interactions between hepatic stellate cells , Kupffer cells and liver sinusoidal endothelial cells by immunofluorescence localization. Conclusion LratCre strain as a spcific reporter mouse for hepatic stellate cells can be uesd to trace hepatic stellate cells.