Sample size was calculated on the basis of the proportion of patients who received at least 50% of the target dose of brand beta-blockers cytotec 200mcg which was around 80% [21–23,26,41]. The estimated proportion of patients who received at least 50% of the target dose of generic beta-blockers was around 40% . This study aimed to compare the proportion of two independent samples at type I error =0.05, type II error =0.10. The calculated sample size for each group =30, and with an estimated occurrence of incomplete data of around 50%, a total sample size of at least 90 patients was needed..
HL-60 cells (2.5×105/ml) in logarithmic growth phase were seeded in 6-well plate. For transfection, 2.5 μg of plasmid pPML(NLS-)-shRNA was diluted in 500 μl Opti-MEM I, which was added to each well. Then, 2.5 μl of PLUS™ Reagent was directly added to dilute DNA. After gentle mixing and incubation for 15 min at room temperature, 13.75 μl of Lipofectamine™ LTX was added into above DNA solution, followed by incubation for 25 min at room temperature. The resultant solution was added to each well. After incubation at 37°C for 6 h, 1 ml of fresh complete medium was added. These transfected cells were named as HL-60/pPML(NLS-)-shRNA. In the negative control group, HL-60/pshRNA-NC cells were processed similarly. At 48 h after transfection, the successfully transfected cells were verified by fluorescent microscopy. The transfection efficiency was expressed as the percentage of GFP-positive cells. There were three groups in this experiment: HL-60 cells group, HL-60/pshRNA-NC group and HL-60/pPML(NLS-)-shRNA group.. However, most patients do not have such clear-cut positive or negative manifestations and thus require testing to evaluate for intra-abdominal injury. Testing options include.
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