Cervical Cancer

Human hepatoma HepG2 cell line was acquired from ATCC China. The cells were cultivated in RPMI 1640 at 37 °C with 5% CO₂ in a moist environment. RPMI was used as per Nasr et al.^{25 (link)}, instead of the regular DMEM media. RPMI 1640 was enriched with 10% fetal bovine serum for cytotoxicity assays along with 2 mM L-glutamine, 50 UI/mL penicillin, and 50 UI/mL streptomycin^{31 (link),32 (link)}. After every 3 days, the medium was replaced. Cell cultures which have attained confluency were dislodged using trypsin, and the cells were planted into 96-well plates at a density of 10⁶ cells/ml. Before being exposed to cubosomes, the cultures were retained at 37 °C for 24 h to allow the cells to achieve confluency and attach to the well plates. These cultured cells were utilized for cytotoxicity and impedance studies.
Human cervical cancer cells HeLa (obtained from ATCC, China), for the fluorescent studies, were obtained from ATCC and cultured in a combination of Eagle’s minimum essential medium (Sigma-Aldrich, China) with 10% fetal bovine serum in an incubator with humidified air or 5% CO₂ at 37 °C. Every 3 days media was renewed, and when reaching 90% confluence cells were passed.

A primary objective of this research was to determine the sensitivity of SaTScan results to the maximum-size parameter. It is important to reiterate that the maximum-size parameter can be either the percentage of the total population at risk or the geographic size of the circle. We use the former because it is the default setting in SaTScan. To understand the sensitivity of SaTScan results to the maximum-size parameter, we ran the SaTScan spatial scan statistic on the cervical cancer dataset fifty times, starting with a maximum-size of 1% and increasing the parameter by an interval of 1 percentage point with each run until reaching the default value maximum-size value of 50%. For each run, we set the number of Monte Carlo replications to 999. The SaTScan software allows users to set the maximum-size in two ways: (1) through restriction of the upper limits of the maximum-size parameter before running the scan or (2) through retention of the default maximum-size of 50%, but selecting for the software to report only circles with a maximum-size value below the desired value. We adopted the second method to avoid pre-selection bias, as recommended by Kulldorff [11 ]. The output for each SaTScan run produces a place ID for each identified cluster; the FIPS code was used as the place ID for this research. The FIPS code at the county level is a five-digit textual code that uniquely identifies counties and county equivalents in the United States. The user can load the files of the fifty SaTScan results into VIT as a batch by specifying the folder where the files are stored in the computer. They are examined in tabular form using the interactive user interface shown in Figure 8. Once the results have been loaded into the VIT's tabular interface, an analyst can interactively investigate each cluster or the individual counties. By selecting one or multiple cluster(s) in the tabular interface, the analyst can select and highlight the cluster(s) on the choropleth map, blurring those counties not included in the selection. We drew a circle around the boundary of each cluster for illustration purposes only; this feature is not implemented in VIT. Specific details (e.g., SMR, p-value, population, etc) about each cluster and individual counties can be interactively retrieved from the map and the tabular interface.