METHODS: Cells were pre-incubated with 32µM of 15dPGJ2 and stimulated with 1ng/mL of IL-1β as an in vitro model of inflammation. Western immunoblotting was used to detect phosphorylated p-65 and phosphorylated c-Jun as markers of NF-κB and AP-1 activation, respectively. mRNA expression of the pro-inflammatory cytokines IL-6, IL-8, and TNF-α was examined, and protein expression of COX-2 and PGE2 were detected by western immunoblotting and ELISA respectively. Myometrial contractility was examined ex-vivo using a myograph.
RESULTS: 15dPGJ2 inhibited IL-1β-induced activation of NF-κB and AP-1, and expression of IL-6, IL-8, TNF-α, COX-2 and PGE2 in myocytes, with no effect on myometrial contractility or cell viability. Despite inhibiting IL-1β-induced activation of NF-κB, expression of IL-6, TNF-α, and COX-2, 15dPGJ2 led to activation of AP-1, increased production of PGE2 and increased cell death in VECs and AECs.
CONCLUSION: We conclude that 15dPGJ2 has differential effects on inflammatory modulation depending on cell type and is therefore unlikely to be a useful therapeutic agent for the prevention of preterm birth.
METHODS: Reverse-transcription-polymerase chain reaction was employed to measure the expression of plasmacytoma variant translocation 1 (PVT1), microRNAs (miRNAs), and SIRT3, and the dual-luciferase assay was used to determine their interaction. Electron microscopy observes autophagosomes, green fluorescent protein-microtubule-associated protein 1 light chain 3 (GFP-LC3) staining, and immunoblot analysis with antibodies against LC3，beclin-1, and P62 were conducted to measure autophagy. Cellular senescence was determined using immunoblot analysis with anti-phosphorylated retinoblastoma and senescence-associated β-galactosidase staining.
RESULTS: Women with higher estrogen levels (during the 10-13th day of the menstrual cycle or premenopausal) exhibit markedly higher serum levels of PVT1 than women with lower estrogen levels (during the menstrual period or postmenopausal). The dual-luciferase assay showed that PVT1 acts as a sponge for miR-31, and miR-31 binds to its target gene, SIRT3. The 17β-E2 treatment increased the expression of PVT1 and SIRT3 and downregulated miR-31 expression in human umbilical vein endothelial cells (HUVECs). Consistently, PVT1 overexpression suppresses miR-31 expression, promotes 17β-E2-induced autophagy, and inhibits H2O2-induced senescence. miR-31 inhibitor increases SIRT3 expression and leads to activation of 17β-E2-induced autophagy and suppression of H2O2-induced senescence.
CONCLUSION: Our findings demonstrated that 17β-E2 upregulates PVT1 gene expression and PVT1 functions as a sponge to inhibit miR-31, resulting in the upregulation of SIRT3 expression and activation of autophagy and subsequent inhibition of H2O2-induced senescence in HUVECs.
METHODS: Retrospective review of 119 consecutive paediatric patients referred for 18F-FDG-PET/CT at the Department of Nuclear Medicine of the National Cancer Institute, Putrajaya. All had DRE and underwent evaluation at the Paediatric Institute, Hospital Kuala Lumpur. Visually detected areas of 18F-FDG-PET/CT hypometabolism were correlated with clinical, MRI and VEM findings.
RESULTS: Hypometabolism was detected in 102/119 (86%) 18FFDG- PET/CT scans. The pattern of hypometabolism in 73 patients with normal MRI was focal unilobar in 16/73 (22%), multilobar unilateral in 8/73 (11%), bilateral in 27/73 (37%) and global in 5/73 (7%) of patients; whilst 17/73 (23%) showed normal metabolism. In 46 patients with lesions on MRI, 18F-FDG-PET/CT showed concordant localisation and lateralization of the EF in 30/46 (65%) patients, and bilateral or widespread hypometabolism in the rest. Addition of 18FFDG PET/CT impacted decision making in 66/119 (55%) of patients; 24/73 with non-lesional and 30/46 patients with lesional epilepsies were recommended for surgery or further surgical work up, whilst surgery was not recommended in 11/46 patients with lesional epilepsy due to bilateral or widespread hypometabolism. 25 patients subsequently underwent epilepsy surgery, with 16/25 becoming seizure free following surgery.
CONCLUSION: 18F-FDG-PET/CT has an added benefit for the localization and lateralization of EF, particularly in patients with normal or inconclusive MRI.