ABSTRACT
BACKGROUND: BrdU is a commonly used reagent in cell proliferation assays, and WST-1 measurement is widely used to detect cell viability. However, no previous study has formally reported the combination of the two assays, which may be used to detect the proliferation and viability simultaneously. In this study, we examined the effect of adding BrdU 2 h prior to the WST-1 assay and tried to test the possibility of the combined detection using rat airway smooth muscle cells. RESULTS: The WST-1 measurements obtained from the combined detection were consistent with those obtained from the separate detection, which suggested that the addition of BrdU 2 h prior to the WST-1 analysis did not affect the WST-1 results. The BrdU measurements obtained from the combined detection also demonstrated the same trend as that obtained from the separate detection, and dosages of 200, 400 and 800 ng/ml testing reagent significantly inhibited the proliferation of rat airway smooth muscle cells. CONCLUSIONS: Our study suggests that the BrdU and WST-1 measurements can be applied simultaneously without mutual interference, which may increase the efficacy and consistency of these measurements to a certain extent.
Subject(s)
Animals , Rats , Technology Assessment, Biomedical/methods , Tetrazolium Salts/pharmacology , Trachea/cytology , Bromodeoxyuridine/pharmacology , Myocytes, Smooth Muscle/physiology , Cell Proliferation/physiology , Reagent Kits, Diagnostic , Trachea/growth & development , Enzyme-Linked Immunosorbent Assay , Cell Survival/physiology , Calgranulin B/administration & dosage , Primary Cell CultureABSTRACT
Background The biomeasurement and imageology of retinal nerve fiber layer(RNFL) thickness showed the damage of retinal structure in the early and middle stage of glaucomatous eye,however,the subtle functional damage of glaucoma can not be timely reflected only with automatic static perimeter.Microperimetry is a method of quantitatively assessing mean sensitivity (MS) of macular zone.Objective This study was to evaluate and compare the macular functional change in early and middle stage of primary open angle glaucoma (POAG) and chronic angle-closure glaucoma (CACG) with MP-1 microperimeter.Metbods This trail protocol was approved by Ethic Committee of Peiking University Third Hospital,and written informed consent was obtained from each subject prior to entering the study group.A cross-sectional and case-controlled study was designed.A total of 126 eyes from 126 subjects were enrolled in the trail,including 53 eyes of 53 normal subjects,50 eyes of POAG patients and 23 eyes of CACG patients.A macular 10° program was set with MP-1 microperimetry to record the MS of various subareas.The macula was zoned into central 2°,6°,and 10° visual fields and 4 quadrants in each ring.The MS of different rings and subareas was detected and compared among POAG patients,CACG patients and normal controls.Results The MS values of central 2°,6°,10° and whole macular area were (15.09 ± 3.03),(15.72 ± 3.22),(13.99 ± 3.63) and (14.91±3.07)dB in the POAG group,which were significantly lower than those of (17.29±1.59),(18.05±1.24),(16.76±1.89) and (17.37±1.46)dB in the normal control group (all at P=0.000).The MS values of central 2°,6°,10° and whole macular area was (16.00±2.39),(15.83±2.63),(14.45±3.15) and (15.42±2.54) dB in the CACG group,and the reduced MSs were seen at the 6°,10° rings and whole macular area in the CACG group compared with the normal control group (P =0.004,0.013,0.011).Within the 6° ring,the MS values in the inferotemporal quadrant were declined in the POAG group and CACG group compared with the normal control group (P =0.000,0.022),but the difference was not statistically significant between the POAG group and the CACG group (P =0.311).In addition,the MS value in the inferonasal quadrant was significantly lower than that of the normal control group (P =0.005); while that in the CACG group was not significantly different in comparison with the normal control group (P=0.119).In the POAG group,the MS value of the inferotemporal quadrant was significantly lower than that of the superonasal or superotemopral quadrant (P =0.043,0.016),but no significant differences were found among the 4 quadrants in the CACG group (all at P>0.05).Conclusions The mild damage of retinal function appears in the early and middle stage of POAG and CACG.More serious MS reducing occurs in the inferotemporal and inferonasal quadrants of POAG.