Non-small cell lung cancer (NSCLC) results in high mortality and has gained increasing attention. C-Phycocyanin (C-PC) has been identified as a potential therapeutic inhibitor for NSCLC, but its underlying mechanism remains obscure. The gene expression of the long noncoding RNA neighbour of BRCAI RNA 2 (NBR2) in NSCLC cells was evaluated by quantitative reverse transcription-PCR. The cell capacity for proliferation and migration was examined by EdU and wound-healing assays. Furthermore, the viability and apoptosis of cells was measured with CCK-8 and annexin V/PI, respectively. Next, the protein level of activation of adenosine monophosphate- activated protein kinase and the rapamycin kinase (mTOR) signalling pathway-associated molecules was evaluated by western blotting. H292 cells were pre-treated with C-PC or transfected with plasmids encoding NBR2 or the shNBR2 plasmid, to over-express or knock down NBR2 expression, respectively. NBR2 expression was robustly down-regulated in NSCLC cell lines compared with a normal cell line (BEAS-2B). NBR2 over-expression inhibited migration and promoted apoptosis of H292 cells. Treatment of H292 cells with C-PC enhanced NBR2 levels in a dose- and time-dependent manner. Downregulation of NBR2 in H292 cells inhibited the activity of C-PC on cell proliferation, viability and clone formation. Further mechanistic investigation showed that the down-regulation of NBR2 abolished the modulatory effects of C-PC on the AMPK/mTOR signalling pathway. In conclusion, C-PC inhibits H292 cell growth by enhancing the NBR2/AMPK signalling pathway.
Cíl práce: Posoudit vliv jednorázové hemodialýzy a dlouhodobé pravidelné hemodialyzační léčby (1 rok) na hladinu C-reaktivního proteinu (CRP) u pacientů s chronickým renálním selháním. Název a sídlo pracoviště: Oddělení klinické biochemie, FN u sv. Anny v Brně. Materiál a metody: Do sledování bylo zavzato celkem 27 nemocných podstupujících pravidelnou hemodialýzu (bikarbonátová hemodialýza, membrána hemophan). Dialýzy probíhaly po dobu 3–4 hodin 3krát týdně. Plazmatická koncentrace CRP, albuminu a diferenciální rozpočet leukocytů byly stanoveny běžnými rutinními metodami. Krev na vyšetření byla odebrána před hemodialýzou a po hemodialýze na počátku sledování a znovu po 12 měsících pravidelné hemodialyzační léčby. K vyhodnocení byly použity párové přístupy statistické analýzy: dvoucestná analýza rozptylu (ANOVA) u parametricky rozložených dat a Wilcoxonův párový test pro neparametrická data. Výsledky: Průměrná koncentrace CRP byla po jednorázové hemodialýze signifi kantně vyšší, než před hemodialýzou (12,3 ± 2,22 a 9,9 ± 2,12 mg/l, p < 0,01). Po 12 měsících pravidelné hemodialýzy nebyl nalezen signifi kantní rozdíl v hladině CRP ve srovnání s hodnotou na počátku sledování (8,6 ± 2,83 a 9,9 ± 2,12 mg/l, p = 0,191). Závěr: Výsledky podporují hypotézu, že aktivaci mediátorů zánětu (CRP) může ovlivnit jednorázová hemodialyzační procedura, hladina CRP však nezávisí na délce trvání chronické hemodialyzační léčby., Objective: To identify if a single hemodialysis session or long-term (one year) hemodialysis treatment infl uences serum C-reactive protein (CRP) level in patients with chronic renal failure. Settings: Department of Clinical Biochemistry, St. Anna’s Faculty Hospital, Brno, Czech Republic. Methods: A total of 27 patients on maintenance hemodialysis (bicarbonate hemodialysis with hemophan membrane) were included in the study. Their chronic dialysis protocol consisted of 3-4 hours sessions three times a week. Plasma CRP concentration, blood leukocyte differential counts and albumin were determined by routine methods. Blood samples were taken before and after a single hemodialysis sessions at the start of the study, and after 12 months of regular hemodialysis treatment. Two-factor analysis of variance in parametrically ordered data and the Wilcoxon paired test for non-parametrically ordered data were used. Results: The mean CRP level after a 4-hour hemodialysis session was signifi cantly higher than that before hemodialysis (12.3 ± 2.22 and 9.9 ± 2.12 mg/l, p < 0.01). No signifi cant changes in mean CRP levels were found after 12 months of regular hemodialysis treatment in comparison with CRP levels at the start of the study (8.6 ± 2.83 and 9.9 ± 2.12 mg/l, p = 0,191). Conclusion: Our study corroborates the hypothesis that the hemodialysis procedure itself can affect infl ammation markers (CRP), but CRP level does not depend on the length of time of regular hemodialysis treatment., Soška Vladimír, Sobotová D., and Lit.: 23
A single random oligonucleotide 3H primer has been previously applied in random-amplfied-polymorphic-DNA (RAPD)-PCR to distinguish stocked bacteria E. coli within a cocktail mixture also containing Enterococcus faecalis, Bifidobacterium longum and Ruminococcus gnavus. In this study, we demonstrate that a 702 base pair (bp) gene fragment can be amplified as a unique pattern by RAPD-PCR using a 3H primer in human faeces containing E. coli. This unique 702 bp amplicon contained a 687 bp gene fragment identified as the C-terminal region of the glutamate-ammonia-ligase adenyltransferase (glnE) gene of E. coli. By high-resolution melt (HRM) analysis, a mean melt-curve temperature of this 702 bp amplicon was determined to be approximately 88.1 ± 0.22 degrees Celsius (°C). A combination of RAPD with HRM in one single reaction based on this amplicon can achieve
semi-quantitative detection of up to 102 CFU/ml of E. coli. To increase the signal intensity of HRM, a primer pair capable of screening E. coli directly from fresh human faeces was re-designed from the 687 bp gene segment, giving a mean peak melt-curve temperature at 88.35 ± 0.11 °C. Finally, single-nucleotide polymorphisms of this 687 bp gene segment were analysed for pathogenic E. coli strains, including UMN026, O83:H1, O104:H4, O157:H7 and O169:H41. We conclude that this 687 bp segment of the glnE gene has a high potential for screening of human faecal E. coli, including pathogenic strains, in contaminated food and water. and Corresponding author: Ko-Tung Chang