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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.
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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 .
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OBJECTIVE:To establish a method for determining recombinant human calmodulin B subunit(rhCNB)in rat plas-ma,and study its pharmacokinetics characteristics. METHODS:ELISA double-antibody sandwich method was adopted. 1 μg/ml rhCNB monoclonal antibody mAb was wrapped,added to the to-be-test sample,rhCNB polyclonal antibody pAb(dilution ratio of 1∶5 000)and HRP-labeled conjugate of anti-IgG(dilution ratio of 1∶10 000)were added. Using tetramethylbenzidine for develop-ing,microplate reader was conducted in wavelength of 450 nm to determine the absorbance value(OD value)and plasma concen-tration of 6 rats after 2,15,30,60,120,240,480,720 min of iv 2.5 mg/kg rhCNB,and the pharmacokinetic parameters were calculated by BAPP 3.0 software. RESULTS:The linear range of rhCNB were 0.195-12.5 ng/ml(r2=0.995 0),lower limit of quan-titation was 0.195 ng/ml,accuracy were 97.300%-103.622%(RSD<7.5%,n=6);RSDs of within-batch,inter-batch,freezing and thawing 3 times were no higher than 8.5%(n=6,18,15). rhCNB pharmacokinetics characteristics in rat fitted to two-com-partment model,AUC0-720 min was 173.038 mg·min/L and t1/2 was 94.62 min. CONCLUSIONS:The established method has high specificity and sensitivity,good accuracy and precision,which can be used for rhCNB quantitative detection and pharmacokinetics study in biological samples.
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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.
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Objective:To investigate the antitumor activity of the compound HS-4 and the action mechanism.Methods:MTT method was used to testin vitroantitumor activity of the compound HS-4. Orthotopic xenotransplantation tumor model of liver cancer was established in nude mice, and,in vivoantitumor activity of compound HS-4 was tested with a small animal in-vivo imaging system. Sequencing of small RNA library and RNA library was performed in HS-4 treated tumor cell group and control group to investigate the anti-cancer mechanism of HS-4 at level of functional genomics, using high-throughput sequencing technology. Results:HS-4 was found to have relatively highin-vitro antitumor activity against liver cancer cells, gastric cancer cells, renal cancer cells, lung cancer cells, breast cancer cells and colon cancer cells. The IC50 values against SMMC-7721 and Bel-7402 of liver cancer cells were 0.14 and 0.13 nmol/L respectively, while the IC50 values against MGC-803 and SGC-7901 of gastric cancer cells were 0.19 and 0.21 nmol/L, respectively. It was demonstrated that HS- 4 possessed a better therapeutic effect in liver cancer.Conclusions: A new reliable orthotopic xenotransplantation tumor model of liver cancer in nude mice is established. The new compounds HS-4 was found to possess relatively highin vivo andin vitroantitumor activity against liver cancer cells.
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Objective To evaluate the antitumor effect of corosolic acid and its impact on CAM and YSM angiogenesis.Methods The effects of CRA on A549 proliferation was studied by MTT method in vitro.Tumor-bruden nude mice model were established by injecting A549 lung cancer cell marked with bioluminescent to nude mice,and the growth of tumor in mice were detected by IVIS small animal in vivo imaging system.Experiment of chicken embryos eggs are used to observe the role of drugs on CAMand YSMblood vessels. Results The value of IC50 of CRA for A549 in vitro was 26.8μg/mL.A tumor-burdened animal experiment results showed that the CRA to A549 solid tumor has a certain therapeutic effect.CAM and YSM blood vessels of chicken embryos eggs treated with CRA were decreased significantly than negative control group.Conclusion CRA has certain therapeutic effect for A549,which may be related to the inhibition of angiogenesis in tumor tissues.