METHODS: In this study, we established persistently NDV-infected EJ28 bladder cancer cells, designated as EJ28P. Global transcriptomic analysis was subsequently carried out by microarray analysis. Differentially expressed genes (DEGs) between EJ28 and EJ28P cells identified by the edgeR program were further analysed by Gene Set Enrichment Analysis (GSEA) and Ingenuity Pathway Analysis (IPA) analyses. In addition, the microarray data were validated by RT-qPCR.
RESULTS: Persistently NDV-infected EJ28 bladder cancer cells were successfully established and confirmed by flow cytometry. Microarray analysis identified a total of 368 genes as differentially expressed in EJ28P cells when compared to the non-infected EJ28 cells. GSEA revealed that the Wnt/β-catenin and KRAS signalling pathways were upregulated while the TGF-β signalling pathway was downregulated. Findings from this study suggest that the upregulation of genes that are associated with cell growth, pro-survival, and anti-apoptosis may explain the survivability of EJ28P cells and the development of persistent infection of NDV.
CONCLUSIONS: This study provides insights into the transcriptomic changes that occur and the specific signalling pathways that are potentially involved in the development and maintenance of NDV persistency of infection in bladder cancer cells. These findings warrant further investigation and is crucial towards the development of effective NDV oncolytic therapy against cancer.
Design: Qualitative study using focus group discussions. Participants' responses were audio recorded, transcribed, grouped under various domains and listed out and analysed.
Setting: A private medical college in Perak state, Malaysia.
Participants: Forty-six medical students from years 2 to 5 were included. Eight focus groups were formed with two focus groups from each academic year (six students each in seven groups and four students in one group). Students were informed through their respective student leader of each year and received a participant information sheet and an informed consent form which were completed and returned if they decided to participate in the focus group discussions.
Results: The participants had different levels of understanding of primary care depending on their level of exposure to primary care. Senior students with more exposure had a better understanding about primary care and its services. Attractive factors towards choosing primary care as a career included short working hours with a more balanced family and social life, being able to treat patients as a whole with continuity of care and closer relationship with patients. Unattractive factors included routine, unchallenging and boring practice, poor salary, work overload and administrative work in government clinics, being less recognised by other specialties; and the poor perception by other doctors that those pursuing primary care were not 'brilliant enough' for more 'sophisticated disciplines like surgery or paediatrics'.
Conclusion: This study showed that the medical students' level of exposure to primary care played a crucial role in determining their understanding of primary care practice and their choice of career in primary care. Issues to be addressed include remuneration, workload and the prejudice against primary care as a career pathway. Suggestions included introducing early exposure to fun and challenging primary care postings in the medical curriculum and producing well trained, skilled and enthusiastic role models.