1.Distribution of autofluorescent microorganisms in the rat intestine
Shuhong TIAN ; Richao WANG ; Min XIAO ; Jian FU
Acta Laboratorium Animalis Scientia Sinica 2017;25(1):97-101
Objective To study the distribution of intestinal autofluorescent microorganisms in the rat intestine at different developmental stages. Methods The distribution of intestinal autofluorescent microorganisms in rat intestine at va-rious developmental stages was tested and evaluated using a small animals living imaging system. First, standard E. coli strain was tested by fluorescence detection in vitro. Then, the distribution of E. coli under the same test conditions was tested. The intestinal autofluorescent bacteria distribution was detected in the SD rats at 3 days,14 days and 60 days of age. After expanding the range of excitation wavelength fluorescence detection,removing the background of fluorescence feed and feces and other foreign autofluorescent substances. Results E. coli can be excited in the range of 485 -535 nm wave?length and to emit fluorescence. E. coli mainly existed in the stomach and only a few E. coli were found in the ileum of 3?days old SD rat. . In the 14?days old rats, E. coli mainly existed in the stomach and cecum, and only a few E. coli were found in the ileum. In the 60-days old SD rats, E. coli mainly existed in the ileum, and only a few E. coli were found in the colon, cecum and jejunum. After the expansion of the excitation light wavelength range of fluorescence detection, E. co?li were observed mainly in the ileum, and only a few E. coli were found in the stomach in 3?days old SD rat. E. coli mainly existed in the stomach, then the cecum and only a few E. coli were found in the ileum and jejunum in 14-days old SD rats. E. coli could be found in the whole intestinal system but mainly in the ileum and cecumin of the 60-days old rats. Conclu?sions Examining the intestinal autofluorescent microbes with the small animal in vivo imaging system can be helpful and make guidance to study the distribution of intestinal microbes in the host at different developmental stages, and to provide a basis for studying the relationship of intestinal microbes with its host and the gastrointestinal drug administration.
2.Application of tissue spontaneous fluorescence in draw focal cerebral ischemia reperfusion injury of rats
Shuhong TIAN ; Richao WANG ; Guilan XING ; Jian FU
Chinese Pharmacological Bulletin 2015;(10):1473-1476
Aim To discuss the application of tissue spontane-ous fluorescence for draw focal cerebral ischemia reperfusion in-jury in rats based on specific fluorescence detection technology. Methods The change of spontaneous fluorescence WAS COM-PARED between the brains of normal rats and those of rats with draw focal cerebral ischemia-reperfusion injury and an quantita-tive analysis was then made. Result The results showed that spontaneous fluorescence of brain tissue for focal cerebral ische-mia reperfusion injury changed significantly. Spontaneous fluo-rescence signal of injury considerably enhanced. The fluores-cence signal which was quantified by IVIS had significant statisti-cal significance compared with normal brain tissue, P <0. 05. Conclusion Our research shows that spontaneous fluorescence of brain tissue enhances obviously after focal cerebral ischemia-reperfusion injury. Our research provides a method for the re-search and evaluation of focal cerebral ischemia-reperfusion inju-ry model in rats.
3.Methodological evaluation of rhCNB in long-tailed macaque sera detected by Enzyme-linked Immunosorbent Assay(ELISA)
Jiping SHAO ; Caiyun ZHANG ; Buwei HU ; Xueli XIE ; Shuhong TIAN ; Richao WANG ; Daolong HUANG ; Jian FU
Chinese Journal of Immunology 2016;32(4):528-531
Objective:To validate an enzyme linked immunosorbent assay (ELISA) method for the quantification of rhCNB in long-tailed macaque sera.Methods: The linear,sensitivity,accuracy,precision and recovery were determined using ELISA.Results:The present ELISA method had high linearity within 0.195 ng/ml-12.5 ng/ml,the working curve of rhCNB was Y=15.1X-0.26, R2=0.996 8 , the method showed good sensitivity of 0.195 ng/ml, the accuracy were in the range of 91.9%-108.8%, and the Coefficient of variation ( CV) for inter-assay were 3.55%,1.39%and 4.71%,the intra-assay were 1.59%,3.2%and 3.8%,all less than 10%, the recoveries were in the range of 88.5% -108.3%, <110% .Thus the method was coincidence with requirement.Conclusion:Double antibody sandwich ELISA assay of rhCNB in long-tailed macaque sera has good sensitivity ,accuracy, precision and recovery and it can be used to measure rhCNB concentration in biological samples .