Effective biomarkers are essential to the early diagnosis of non-small cell lung cancer (NSCLC). Herein, a retrospective study of 49 newly diagnosed and recurrent NSCLC patients, 31 patients with benign pulmonary disease and 24 healthy volunteers was conducted, to evaluate the diagnostic value of circulating rare cells for NSCLC. The expression of circulating tumor cells (CTCs) and circulating tumor-derived endothelial cells (CTECs) in peripheral blood were measured by subtraction enrichment-immunostaining-fluorescence in situ hybridization (SE-iFISH). The level of CTCs (P＜0.001) and CTECs (P＜0.001) was significantly higher in NSCLC group than that in benign pulmonary disease group. The proportion of small CTCs (P＜0.001) and CTECs (P＜0.0001) significantly increased from benign lung disease individuals to NSCLC patients. The AUC of ROC curves of total CTCs and CTECs were 0.815 (95%CI: 0.722~0.907), 0.739 (95%CI: 0.618~0.860), respectively. The cut-off values for discriminating NSCLC with benign lung disease patients were total CTCs 11.5 units/6ml and total CTECs 10.5 units/6ml, with sensitivity and specificity being 67.3% and 83.9%, 77.6% and 77.4%, respectively. When CTCs and CTECs were combined, predictive value significantly increased to 82.6% as measured by the area under the curve. Small CTCs and triploid CTCs had high positive predictive value (PPV) and positive likelihood ratio (LR+) of the diagnosis of NSCLC in early stage. CTCs and CTECs can not only be used as new biomarkers for the diagnosis of NSCLC, but can also improve diagnostic performance of the early stage NSCLC. Moreover, the combined examination of CTCs and CTECs is be superior to the single.

Broomrape (Orobanche cumana), root parasitic weed, negatively affects sunflower, thereby causing severe yield losses and required controls on O. cumana infestation. Brassinosteroids (BRs) plays key roles in plant growth and provides resilience to weed infection. This designed study evaluated the mechanisms by which BRs ameliorates O. cumana infection in sunflower (Helianthus annuus). The pretreatment of sunflower seeds with various concentrations of BRs (1, 10, and 100 nM) were applied with and without O. cumana infestation. Under O. cumana infection, a significant reduction in plant height (38.61%), fresh/dry biomass, photosynthesis and regulation in plant defense (POX, GST), BRs signaling (BAK1, BSK1 to BSK4) and synthesis (BRI1, BR6OX2) associated genes. Moreover, O. cumana elevated the levels of MDA, OH‒, H2O2 and O2•‒ in leaves/roots by 77%/112%, 62%/104%, 57%/98% and 55%/89.4% and caused ultrastructural damages in leaves/roots, suggesting the severe oxidative or cellular damages in sunflower. In response, plants activate activities of SOD and POD enzymes as well as reduced or oxidized glutathione, but unable to stimulate APX and CAT activities. Addition of BRs, especially at 10 nM notably lowered the production of oxidative stress, activated the enzymatic and non-enzymatic antioxidants. The downregulation in the reported genes by BRs attributed to the increased resilience of sunflower (via susceptible reaction). In nutshell, the pretreatment of BRs enhanced the sunflower resistance to O. cumana infection by improving the planta growth and biomass, photosynthesis, activating the antioxidant defense system, lowering the oxidative stress, cellular damages and modulating the expression of BR synthesis and signaling genes.