Long noncoding RNAs (lncRNAs) are emerging as important regulators of numerous biological processes, especially in cancer development. Aberrantly expressed and specifically located in tumor cells, they exert distinct functions in different cancers via regulating multiple downstream targets such as chromatins, RNAs, and proteins. Differentiation antagonizing non-protein coding RNA (DANCR) is a cytoplasmic lncRNA that generally works as a tumor promoter. Mechanically, DANCR promotes the functions of vital components in the oncogene network by sponging their corresponding microRNAs or by interacting with various regulating proteins. DANCR's distinct expression in tumor cells and collective involvement in pro-tumor pathways make it a promising therapeutic target for broad cancer treatment. Herein, we summarize the functions and molecular mechanism of DANCR in human cancers. Furthermore, we introduce the use of CRISPR/Cas9, antisense oligonucleotides and small interfering RNAs as well as viral, lipid, or exosomal vectors for onco-lncRNA targeted treatment. Conclusively, DANCR is a considerable promoter of cancers with a bright prospect in targeted therapy. Approximately 75% of the human genome can be transcribed into RNAs, yet no more than 2% encodes proteins . Non-codDue to the overwhelming quantity of lncRNAs in the human genome and the lack of methods for predicting their functions, massive work is still needed to better understand the lncRNAs. LncRNA differentiation antagonizing non-protein coding RNA (DANCR) can serve as a paradigm for lncRNA investigation. DANCR is a typical oncogenic lncRNA which exerts multiple functions in cancer development. It has also demonstrated its value in cancer therapy as a promising target .This article summarized the functional properties and mechanisms of DANCR in tumor progression. The general targeted therapies against onco-lncRNAs were introduced and the specific methods to target DANCR were discussed. This article can provide referential value for latter onco-lncRNA investigations and insights into the clinical treatment against cancers caused by aberrant expression and regulation of lncRNAs.DANCR also named as Anti-Differentiation Non-coding RNA (ANCR), Small nucleolar RNA host gene protein 13(SNHG13), upregulating transcripts 2 (AGU2) is an oncogenic lncRNA located on chromosome 4 . It was The discovery that DANCR over-expressed in colorectal cancer promoted tumor progression and predicted poor prognosis introduced the study of DANCR into the field of cancer . DANCR cIn the wake of the deepening of the research, DANCR was found to be aberrantly expressed in numerous cancers and exerted multiple regulating functions. The underlying mechanisms were also being constantly elucidated.DANCR is a typical oncogenic lncRNA, overexpressed in various tumor cells. It can regulate the progression of numerous cancers by modulating the expression or activity of downstream targets and consequently promoting hallmarks of cancer including cell proliferation, cell motility, tumor angiogenesis, and cell viability.In hepatocellular carcinoma, it is observed that down-regulated DANCR expression can boost cell apoptosis and cell cycle block in G1 which underlies that DANCR can promote tumor cell proliferation by inhibiting cell apoptosis and facilitating growth inhibition evasion . DANCR oCell motility refers to the metastasis and invasion properties of cells and malignant cells to vital organs and it is the major cause of cancer deaths . DANCR cDANCR promotes the motility of nasopharyngeal carcinoma cells by stabilizing hypoxia inducible factor-1α, the subunit of Hypoxia inducible factor 1 (HIF-1) which are transcription factors that can activate several hypoxia-responsive genes in response to hypoxia microenvironment and promote the metastasis and invasion of cancer cells , 25. MatTumor angiogenesis is one of the hallmarks of cancer which provide essential nutrients and oxygen to proliferating cancer cells . DANCR cThe innate or acquired chemo-resistance of tumor cells is a main problem in the clinical cancer therapy. DANCR promotes the docetaxel resistance in prostate cancer cells by mediating the upregulation of multidrug resistance associated protein (MDR) and p-glycoprotein (P-gp), which bump many chemotherapeutic drugs out of the cells and limit the intracellular drug dosage to sub-therapeutic levels , 35. DANGenerally, DANCR exerts multiple regulatory functions on tumor progression of numerous cancers, revealing its role as an important pro-tumor regulator . And notLncRNAs perform their multiple regulatory functions on tumors by interacting with numerous downstream targets . The dowEZH2 is a common target protein of DANCR. Mechanically, EZH2 enables the silencing of gene expression by catalyzing histone H3 trimethylation at lysine 27 (H3K27me3) in target gene promoters . DANCR cMicroRNAs are small regulating RNAs (−22 nt in length) derived from the short hairpin regions of RNA transcripts and they induce cleavage and suppression of translation via complementary binding to target RNAs . DANCR cIn this section, the role of DANCR in particular cancers is discussed so as to clarify the molecular mechanisms of DANCR in a more specific way and instruct the therapy of DANCR aberrantly-expressed cancers .DANCR acts as a miRNA sponge in colorectal cancer (CRC) cells, which is the most common role that DANCR plays in cancer development. Harboring the same miR-577 binding site, DANCR and heat shock protein 27 (HSP27) mRNA competitively bind to miR-577. The overexpression of DANCR in CRC cells alleviates the degradation impact of miR-577 on HSP27, which contributes to cell proliferation and metastasis .DANCR activated by SALL-4   mediatesHIF-1α, generally known as the core regulator of the hypoxic response, has been validated as a critical positive regulator of nasopharyngeal carcinoma (NPC) cell metastasis. DANCR can combine the NF90 (nuclear factor 90)/NF45 (nuclear factor 45) complex with its AU-rich elements (AREs) in 3′ region, which stabilizes HIF-1α mRNA and upgrades the motility of NPC cells .In triple-negative breast cancer (TNBC), DANCR functions by mediating protein assembly and modification. DANCR can bind with the phosphorylation site of RXRA, recurring glycogen synthase kinase 3 beta (GSK3β) to sponge RXRA and promote RXRA ser phosphorylation. The structural change in RXRA suppresses its interaction with the promoter of phosphatidylinositol-4,5-biophosphate 3-kinase catalytic subunit alpha (PIK3CA), thereby enhancing PIK3CA in both mRNA and protein levels. Consequently, protein PIK3CA activates the P13K/AKT pathway, leading to the proliferation and tumor growth of TNBC .Bladder cancer (BC) whose proper diagnosis and efficient treatment are current challenges in urology, is the most frequent malignancy of the urinary tract . The folSimilar to CRC, oncogenic DANCR within hepatocellular carcinoma (HC) has also been found to have multiple regulating targets. KLF12 (Krueppel-like factor 12) can be positively regulated by DANCR through miR-216a-5p sponging, leading to malignant progression . DANCR cWe can conclude that DANCR can regulate the signaling pathways by alleviating miRNA's inhibitory functions on target RNA, combining with proteins and guiding them to modulate the expression or the stability of RNAs or facilitating the integration and subsequent modifying of proteins. Moreover, DANCR can manipulate more than one target in cancer and join in multiple pathways. As we can see, due to the diverse mechanisms of DANCR and limited research on it, it is hard to foresee the unknown mechanisms and functions of DANCR in cancers. However, lncRNA's collective involvement in the pathways of numerous cancers does make it a promising target for cancer therapy.The potency of lncRNA-based cancer therapy has been pointed out in previous years. Targeting lncRNAs for cancer treatment is a novel and promising therapy due to their collective therapeutic efficacy, high specificity, and few side effects . The posThe CRISPR/Cas9 technology can be used to target the endogenous loci of lncRNAs and restrict their transcription . The CRIin vivo. Combined with their relatively high charge and hydrophilicity (Antisense oligonucleotides (ASOs) can be used in lncRNA-targeted therapy. They are synthetic and modified oligonucleotide single strands that can complementarily bind to certain sequences of their target RNAs. ASOs exert their inhibiting effect mainly by recruiting ribonuclease H (Rnase H) to degrade the targets. They comprise complementary DNA single strands to form DNA/RNA duplexes for Rnase H recognition and later cleavage . Vulnerahilicity , all thehilicity  that invhilicity . In thishilicity . Additiohilicity . Howeverhilicity , which ihilicity . Uniformhilicity . Among ahilicity . LNA canhilicity . The cenhilicity . Both mihilicity . And to hilicity .We can also use small interfering RNAs (siRNAs) to target lncRNAs. In 2018 patisiran became the first approved RNAi drug to remedy hereditary transthyretin amyloidosis . SiRNAs Viral vectors were one of the earliest engineered nanoscale delivery systems used for transporting RNA products to targeted tissues. Viral vectors are known for their efficient transfer of codes to the cell interior and capability for escaping immunosurveillance by infected cells . In tumoCompared to viral vectors, non-viral vectors are of lower transfection efficiency . Howeverin vivo  finin vivo . HEK293Tin vivo ; (b) train vivo ; (c) isoAlvarez-Erviti et al. pioneered the practice of applying engineered exosomes to deliver siRNA. They build neuronal cell-targeted exosomes and use them to pass through the blood-brain barrier and treat Alzheimer's disease . A recenTo date, researches on targeting DANCR for cancer therapy remains limited. A previous study presented that the relative enrichment of the enzymes responsible for RNA degradation vary between cellular compartments, so the location of lncRNA can impact the suppressing efficacy of the molecular drugs on it. Comparatively, ASO is more capable of clearing the nuclear lncRNAs while RNAi have a better suppressive effect on lncRNAs in cytoplasm . ReferriThe review has shown the vital research value of DANCR. DANCR is also a critical oncogenic regulator which presents an increasingly important status in cancer study. It can regulate hallmarks of various cancers, indicate their progression and clinical outcomes and serve as a novel target for cancer targeted treatment. Researches on DANCR remain limited and there is an urgent need for further study on this critical onco-lncRNA.The recent progress on RNA interaction identification method includes the refined variants of immunoprecipitation techniques  and new high-throughput RNA interactome analysis methods  . WithoutS-JJ conceived, wrote the manuscript, and completed the figures and tables. M-ZJ revised the manuscript. W-LJ and B-RX conceived, organized, and edited the text.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Ischemic conditioning is defined as a transient and subcritical period of ischemia integrated in an experimental paradigm that involves a stimulus of injurious ischemia, activating endogenous tissue repair mechanisms that lead to cellular protection under pathological conditions like stroke. Whereas ischemic pre-conditioning is irrelevant for stroke treatment, ischemic post-conditioning, and especially non-invasive remote ischemic post-conditioning (rPostC) is an innovative and potential strategy for stroke treatment. Although rPostC has been shown to induce neuroprotection in stroke models before, resulting in some clinical trials on the way, fundamental questions with regard to its therapeutic time frame and its underlying mechanisms remain elusive. Hence, we herein used a model of non-invasive rPostC of hind limbs after cerebral ischemia in male C57BL6 mice, studying the optimal timing for the application of rPostC and its underlying mechanisms for up to 3 months. Mice undergoing rPostC underwent three different paradigms, starting with the first cycle of rPostC 12 h, 24 h, or 5 days after stroke induction, which is a very delayed time point of rPostC that has not been studied elsewhere. rPostC as applied within 24 h post-stroke induces reduction of infarct volume on day three. On the contrary, very delayed rPostC does not yield reduction of infarct volume on day seven when first applied on day five, albeit long-term brain injury is significantly reduced. Likewise, very delayed rPostC yields sustained neurological recovery, whereas early rPostC  results in transient neuroprotection only. The latter is mediated via heat shock protein 70 that is a well-known signaling protein involved in the pathophysiological cellular cascade of cerebral ischemia, leading to decreased proteasomal activity and decreased post-stroke inflammation. Very delayed rPostC on day five, however, induces a pleiotropic effect, among which a stimulation of angioneurogenesis, a modulation of the ischemic extracellular milieu, and a reversal of the stroke-induced immunosuppression occur. As such, very delayed rPostC appears to be an attractive tool for future adjuvant stroke treatment that deserves further preclinical attention before large clinical trials are in order, which so far have predominantly focused on early rPostC only. With the recent success of endovascular strategies to avoid stroke induced brain injury , treatmeIschemic conditioning is an experimental strategy that has been studied for decades, using a variety of animal models including but not limited to stroke models . As suchTo this date, ischemic post-conditioning has been shown to be neuroprotective in a variety of experimental stroke models , albeit Whereas immediate ischemic post-conditioning of the brain itself is impractical in clinical settings for obvious reasons, the concept of remote ischemic post-conditioning (rPostC) has gained a substantial amount of attraction. Indeed, rPostC in general and hind limb rPostC in particular has been shown to be neuroprotective, albeit survival periods of these studies did not extend 2 weeks, giving insights in some underlying cellular signaling pathways only ,b. AlthoAll experiments were carried out according to the guidelines for the care and health of animals and were approved by local authorities. Experimental procedures followed the ARRIVE recommendations. Mice were randomly allocated to the experimental treatment groups. Surgeons and analysts were different from each other so that analysts were blinded to the treatment paradigms. Survival periods are given in Supplementary Table in situ for 60 min under constant laser Doppler flow control. The body temperature was continuously measured using a rectal feedback probe and a heating pad, keeping the body temperature between 36.5°C and 37°C. Consequently, this setting allows for brain infarcts affecting the striatum and part of the cortex.Male C57BL6 mice (24–28 g) were exposed to 60 min of middle cerebral artery occlusion (MCAO) as previously described , followeInduction of rPostC was essentially performed as previously described with some modifications . Non-invBrain injury at acute and subacute time points was assessed using triphenyltetrazolium chloride (TTC) staining on 2-mm-thick brain slices. In these slices, infarct volume was outlined and brain edema was calculated as relative increase of the ipsilateral compared to contralateral hemispheric volume. For long-term assessment of brain injury, animals were sacrificed on day 84 after MCAO for which mice were transcardially perfused with 4 % paraformaldehyde in 0.1 M phosphate-buffered saline (PBS). Thereafter, brains were removed, and 20 μm coronal cryostat sections were collected. The latter were used for immunohistochemistry for the neuronal marker NeuN, which was detected by a monoclonal mouse anti-NeuN antibody . Quantitative analysis of the density of surviving neurons in the ischemic striatum was done within four regions of interest in three sections per animal at AP + 0.14 mm, ML ± 1.5–2.25 mm, and DV -2.5–3.25 mm from bregma.+ cells, BrdU+/Dcx+ neurons, BrdU+/NeuN+ neurons, and BrdU+/CD31+ endothelial cells was evaluated as described for NeuN immunohistochemistry using the same coordinates and regions of interest.Neuroregeneration as indicated herein by endogenous neurogenesis and angiogenesis, was analyzed 3 months after stroke induction. As such, mice received intraperitoneal injections of bromodeoxyuridine  on days 8–28. Following transcardial perfusion with 4% paraformaldehyde in 0.1 M PBS on day 84, immunohistochemical analysis was performed in coronal cryostat sections using the following primary antibodies: monoclonal mouse anti-BrdU , monoclonal rat anti-BrdU , polyclonal goat anti-doublecortin  and monoclonal mouse anti-NeuN  and monoclonal rat anti-CD31 . Following detection with appropriate secondary antibodies, the density of proliferating BrdUAll mice received training 1–2 days before MCAO in order to ensure proper test performance in the rota rod test, the tight rope test, the corner turn test, and the balance beam test. These tests were performed as previously described . The rotFor some experiments, stereotactic injections prior to induction of MCAO were performed in order to evaluate whether or not Hsp70 is involved in the aforementioned neuroprotection on day 3 induced by rPostC. As such, Hsp70 was knocked down using an Accell SMARTpool mouse Hsp70 small interfering RNA (siRNA) , which was intracerebrally administered into the striatum 3 days before stroke as previously described . For conTris-Hcl, 145 mM NaCl, 10 mM EDTA, and 0.5% Triton X-100 at pH 7.5). The chymotrypsin-like activity of the proteasome was measured using Suc-LLVY-AMC  as a substrate that was incubated with 90 μl of reaction buffer containing 50 mM Tris, 20 mM KCl, 1 mM magnesium acetate, 2 mM dithiothreitol (DTT), 1 mM leupeptin, 1 μg/ml aprotinin (Sigma-Aldrich) and 1 mM PMSF . The extent of substrate cleavage was evaluated at 37°C in a fluorescence microtiter plate reader at λexc. = 355 nm and at λem. = 460 nm. Since Suc-LLVY is not exclusively cleaved by the proteasome, specificity was increased by adding the proteasome inhibitor MG-132  at the beginning of the measurement. Proteasome activities were given as arbitrary units per min per mg protein. Protein concentration was measured using the Bradford assay.Assessment of proteasome activity was performed as described previously . BrieflyThe concentration of brain-derived neurotrophic factor , glial cell line-derived neurotrophic factor , vascular endothelial growth factor , nerve growth factor , basic fibroblast growth factor  and epidermal growth factor  was measured by enzyme linked immunosorbent assay (ELISA) in left (ischemic) hemispheres 84 days after MCAO as described before .4Cl, 10 mM KHCO3 and 3 mM EDTA. For flow cytometry measurements of brain tissue samples, ischemic left hemispheres were mechanically homogenized in a buffer of collagenase type XI (125 U/ml), hyaluronidase (60 U/ml) and collagenase (450 U/ml) in Ca2+/Mg2+ supplemented PBS . Samples were incubated with the antibody in question as described afore . For comparisons between two groups, statistical comparisons were done using the Student Experimental stroke paradigms analyzing the effect of rPostC on brain injury are usually restricted, as they apply one experimental paradigm with observation periods no longer than 2 weeks only. Hence, we systematically analyzed three different experimental paradigms that differed between the start of the first induction of rPostC. Indeed, early rPostC reduces infarct volumes 3 days after stroke when applied within 24 h after stroke onset Figures . The redInduction of early rPostC during the acute phase of the disease, i.e., 12 and 24 h after stroke onset, yields transient neuroprotection only that is lost during the observation period of 3 months  is mediated via Hsp70 which in turn inhibits proteasomal activation, yielding transient reduction of brain injury. On the contrary, very delayed rPostC reverses peripheral post-stroke immunosuppression which implies a modulation of the extracellular milieu, stimulating neuroregenerative processes that orchestrate sustained neurological recovery.Direct ischemic post-conditioning has been shown to induce both neuroprotection and neurological recovery in a variety of experimental stroke models, including focal and global cerebral ischemia . AlthougNot only in light of its potential therapeutic relevance but also due to its practicability has ischemic rPostC been thoroughly studied in a variety of experimental models. The latter implies experimental studies on the kidney, the liver, the lung, the heart, and the brain, to name but a few . The mecThe precise cellular and molecular mechanisms that are involved are neither known for pre-conditioning, per-conditioning, or post-conditioning of the brain, may it be direct conditioning or rPostC. Nevertheless, a great deal of evidence suggests that these different concepts of conditioning share some general signaling pathways . Indeed,Induction of early rPostC does not yield sustained neuroprotection during the observation period of 3 months which is in line with previous data from our group, demonstrating that direct cerebral post-conditioning induces transient neuroprotection only . LikewisThe concept of remote conditioning in general and the concept of rPostC in particular are interesting concepts for adjuvant treatment paradigms of different diseases. Indeed, first clinical trials have been performed, suggesting possible beneficial effects after remote conditioning . Yet, thTD designed and reasoned the study, performed the experiments, wrote the manuscript, and analyzed the data. BZ performed the experiments and analyzed the data. BK, FJ, XZ, and VV performed the experiments. AM designed and reasoned the study. MB and DH designed and reasoned the study, provided financial support, and wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


AGEs induced the calbindin-D28k expression in human renal proximal tubule cells (HK2), but not in mesangial cells. AGEs induced the expression of fibrotic molecules, ECM proteins, epithelial-mesenchymal transition (EMT) markers, and endoplasmic reticulum (ER) stress-related molecules in HK2 cells, which could be inhibited by a receptor for AGE (RAGE) neutralizing antibody. Calbindin-D28k knockdown by siRNA transfection reduced the cell viability and obviously enhanced the protein expressions of fibrotic factors, EMT markers, and ER stress-related molecules in AGEs-treated HK2 cells. Chemical chaperone 4-Phenylbutyric acid counteracted the AGEs-induced ER stress and ECM and EMT markers expressions. Calbindin-D28k siRNA in vivo delivery could enhance renal fibrosis in db/db diabetic mice. These findings suggest that inducible calbindin-D28k protects against AGEs/RAGE axis-induced ER stress-activated ECM induction and cell injury in renal proximal tubule cells.Diabetes-associated advanced glycation end-products (AGEs) can increase extracellular matrix (ECM) expression and induce renal fibrosis. Calbindin-D28k, which plays a role in calcium reabsorption in renal distal convoluted tubules, is increased in a diabetic kidney. The role of calbindin-D28k in diabetic nephropathy still remains unclear. Here, calbindin-D28k protein expression was unexpectedly induced in the renal tubules of  Diabetes is a metabolic disease resulting from β-cell destruction/insulin deficiency type 1 diabetes) or insulin resistance/insulin secretory defect type 2 diabetes) or other specific diabetic types, causing hyperglycemia  and a negative control siRNA  were purchased from Invitrogen . Cells were seeded on 6-well plates and were transfected with scramble or calbindin-D28k siRNA by using Lipofectamine RNAiMAX transfection reagent (Thermo Fisher Scientific). The siRNA/Lipofectamine medium was removed after 6 h of incubation. Cells were then cultured for 48 h at 37 °C. The siRNA sequences targeting calbindin-D28k were as follows: sense 1: 5′-ACU GAC CAC AGU GGC UUC AUA GAA A-3′; antisense 1: 5′-UUUCUAUGAAGCCACUGUGGUCAGU-3′; sense 2: 5′-ACUGAAGGAUCUGUGCGAGAAGAAU-3′; antisense 2: 5′-AUUCUUCUCGCACAG AUCCUUCAGU-3′; sense 3: 5′-GAAGAACAUAAUGGCUUUGUCGGAU-3′; antisense 3: 5′-AUCCGACAAAGCCAUUAUGUUCUUC-3′.db/db mice were randomized into two groups (scramble and siRNA). The siRNA-treated mice received 530 μL of kidney In Vivo Transfection reagent (Altogen Biosystems) in combination with 90 μg CaBP-D28K siRNA. Control animals received 530 μL of In Vivo Transfection reagent in combination with 90 μg scramble siRNA. Diabetic mice were rapidly intraperitoneal (i.p.) injected scramble or CaBP-D28K siRNA, once per 3 days for a total of 6 times. All mice were sacrificed 48 h after last injection. The kidneys were collected for Western blot, Masson’s trichrome staining, and immunohistochemistry assay. For quantification of Masson’s trichrome staining, the collagen levels were analyzed in a blinded fashion with digital image analysis software .The optimized modification and endotoxin free of CaBP-D28K or scrambled siRNAs (Invitrogen) were dissolved in RNase/DNase free water. Male 8-week-old p-value of less than 0.05 is considered to be a significant difference. Statistical analysis was performed by using GraphPad Prism software, version 6.0 .Data are presented as means ± SEM. The difference between control and treatment groups was assessed by one-way analysis of variance (ANOVA) and followed to the Dunnett’s test. A db/db diabetic mice compared to control db/m+ mice. The data were presented in db/db mice was higher than that in control mice . The data were presented in To investigate the roles of the inducible calbindin-D28k in renal proximal tubule cells after AGEs treatment, the expression of calbindin-D28k was interfered by using siRNA transfection. The data were presented in AGE-RAGE signal pathway is known to play an important role in the pathogenesis of diabetic nephropathy . TherefoA previous study indicated that ER stress-related molecules were involved in the AGEs (10–160 μg/mL)-induced glomerular mesangial injury . The rolNext, a chemical chaperone, 4-PBA, was used to investigate the effects of calbindin-D28k on AGEs-induced ER stress and fibrotic signals in HK2 cells. The data were presented in db/db diabetic mice. The data were presented in db/db mice with siRNA in vivo delivery as compared to scramble control. We also tried to magnify the scale to observe the proximal tubules in kidney sections. We found that calbindin-D28k staining was clearly shown in proximal tubules in which the brush borders in some proximal tubule cells were shed . The 20–40% of diabetic patients has been estimated to develop a chronic kidney disease that for a significant number of patients can lead to ESRD . Studiesdb/db diabetic mice. Unexpectedly, calbindin-D28k could be induced in the proximal tubules of the db/db diabetic mouse kidney, but not control mouse kidney. We further used calbindin-D28k siRNA to silence calbindin-D28k expression in the kidneys of db/db diabetic mice and found that CTGF protein expression and renal fibrosis were markedly enhanced. These in vivo results are consistent with the in vitro findings.Interesting, the expression of calbindin-D28k, a high-affinity cytosolic calcium binding protein, has been shown to be increased in the kidney and it was associated with the increase in the fractional excretion for calcium in the distal convoluted tubule of streptozotocin-induced diabetic rats . Calbind2+ balance might cause ER stress and activate UPR including CHOP and IRE-1-associated signals, leading to the cell death or rebuild the normal ER function [Previous studies indicated that ER stress activation is an important feature for the process of recovery, adaptation, and apoptosis during diabetic nephropathy. Several ER stress-associated signals, including CHOP and/or IRE1-mediated JNK pathway, can cause renal cells apoptotic death, prolonged oxidative stress-induced kidney damage and/or kidney fibrosis ,24,25. Cfunction . The altfunction ,31.A previous study has indicated that ER stress inducer can induce EMT in human peritoneal mesothelial cells . ER streShin et al. have indicated that ER stress can serve as a target for ameliorating EMT via Smad2/3-related signals in human peritoneal mesothelial cells . The actdb/db mice. We have also demonstrated that exposure of AGEs significantly induces cell apoptosis via an ER stress-triggered signaling pathway in mesengial cells (MES-13) [Martes foina, the immunohistochemical positive cells for calbindin-D28k were located in the distal tubules, but not in the glomeruli, proximal tubules, and loops of Henle [db/db diabetic mice (a model of Type 2 diabetes). The calbindin-D28k was not located in the diabetic glomeruli. Therefore, we further hypothesized that the inducible calbindin-D28k might be involved in the process of AGEs-induced renal proximal tubule cell injury and fibrosis. The results suggest that inducible calbindin-D28k protects against AGEs/RAGE axis-induced ER stress-activated ECM induction and cell injury in renal proximal tubule cells.The previous studies of reference  and our (MES-13) . We have(MES-13) ,42 and p(MES-13) . Calbind(MES-13) . It has (MES-13) ,45. Karaof Henle . In the of Henle . In the 2+ transporting 1 (PMCA1) is not changed or slightly decreased [2+ transport in renal distal tubular cells; ER stress-evoked ER Ca2+ depletion-disturbed intracellular Ca2+ homeostasis may be involved in the alteration of CALB1 expression. In the present study, we unexpectedly found that AGEs induced calbindin-D28k expression in renal proximal tubular cells (HK2 cells) and calbindin-D28k silencing enhanced the ER stress signaling and the expression of fibrosis-related molecules. However, the role of calbindin-D28k in the regulation of ER Ca2+ levels in renal proximal tubules under AGEs or diabetic condition is still unclear and needs further investigation.Calbindin D28k possesses four high calcium binding sites and consecutively expressed with higher level in the distal renal tubular region to maintain the calcium homeostasis . Chiappiecreased . They suAGEs with various concentrations have been shown to affect the cell growth and function in various cell types. AGEs (AGE-BSA) at 100 μg/mL have been found to reduce the viable cell numbers and induced apoptotic cell death at 250 μg/mL in retinal pericytes . It has db/db diabetic mouse model [Moreover, the loss of proximal tubule brush border has been observed in the kidneys of streptozotocin/nicotinamide-induced type 2 diabetic mouse model  and db/dse model . Therefodb/db diabetic mice. Calbindin-D28k may serve as a therapeutic strategy for the improvement of diabetic nephropathy. However, more evidence from animal studies is needed to confirm the role of calbindin-D28k in diabetic nephropathy in the future. Moreover, further investigation also needs to clarify the role of calbindin-D28k in calcium homeostasis of renal tubules during diabetes.The results of this study showed that the inducible calbindin-D28k plays an adaptive and protective role against AGEs-induced ER stress and the renal fibrosis signals that occurs afterwards in the proximal tubule cells. Calbindin-D28k can be induced in the renal proximal tubules in


During ultra high voltage (UHV) transmission, the discharge caused by high intensity electric fields aggravates the aging process of external insulation materials used for composite insulators. The microstructural characteristics of its base material polymer—methyl vinyl silicone rubber—are the key basis for the performance of insulation materials under electric field exposure. Based on molecular dynamics simulations, a molecular model of methyl vinyl silicone rubber was established. Mechanisms influencing the microstructure evolution under electric fields had been studied at the atomic level. The results showed that the initial reaction characteristics of silicone rubber molecules involve the violent vibrated of all the methyl and vinyl atoms, and shortening of the chemical bonds. The neighboring groups were close to each other and generated different amounts of -Si-Si- bonds. This promoted the helical shrinkage of the molecule, and protrusion of the middle of the molecule which presented an inverted U shape. The high electric field greatly reduced the total energy of molecules, and the potential energy in particular was more severely destroyed, resulting in degradation of its structure. Besides, as the electric field intensity increased, the elastic modulus of the molecule gradually increased. It was shown that high electric fields would make the stiffness of silicone rubber become larger, and the brittleness to become stronger, which reduced the mechanical properties of materials, accelerating its aging. The results provide a theoretical basis for establishing the connection between the micro appearance and macro characteristics of materials, as well as reference values for the optimization of base materials used for making composite insulators. As composite insulators have excellent performance, they have obvious advantages in the external insulation selection for ultra high voltage UHV transmission projects . HoweverIn most cases researchers have studied composite insulators aged by electric field experiments. Reference  shows thThe current studies are mainly focused on discharge aging of insulation materials induced by increasing electric fields. The aging process of materials ranges from quantitative changes to qualitative changes. However, the above studies rarely examined the structural evolution of the material as the electric field intensity increases. When the material is initially placed in an increasing electric field, changing characteristics of internal structure are the early stage of discharge aging and further degradation. The above studies derived and analyzed the aging state of materials from macroscopic characterizations, which are the cumulative result of the material’s micro characteristics, but the changes of the micro characteristics are difficult to observe directly from experiments.β-cristobalite under different electric fields. The influence factors of polymer dynamic stability were studied in [Due to the limitations of experimental means, the molecular simulation method has attracted more attention and has been widely used, especially in the micro characteristics of the material field, where several remarkable achievements have been made. For example, the simulation method has been applied in the cellulose degradation process of oil paper insulation by Liao et al. ,12. Via udied in ,25. The udied in . It is eudied in .The most used base polymer for composite insulators at home and abroad is methyl vinyl silicone rubber (MVQ) . Based oEGauss), the real space , the reciprocal space (Erecip-space) and the corrected energy term (Ecorr)of the Gaussian distribution [As the electric field plays a key role on the properties of silicone rubber, electrostatic interaction is calculated by the Ewald summation method. In the method, each particle interacts with the mirror image of all other particles in the simulation box and all the particles in the surrounding lattice. It is also assumed that each particle is surrounded by the neutralized charge distribution with equal opposite sign charge. Thus, the electrostatic interaction is the sum of the interaction between Gaussian functions  and (3):E is the elastic constant, G is shear modulus, K is bulk modulus, and v is Poisson’s ratio.In the formulas, 3, and a small amount of unsaturated groups are introduced on side chains, such as -CH=CH2. The intermolecular forces are small, and methyl groups arranged outwards can rotate freely, so the silicone rubber has a certain degree of hydrophobicity.The main material of MVQ is polymethylvinyl siloxane, which is a high molecular copolymer of dimethyl siloxane and methylvinyl siloxane chain segments. The molecular structure can be seen in 5. It is impractical to model 400~800 thousand molecules and carry out simulations under different electric field intensities. However, the polymerization degree was too small to represent the basic characteristics of the polymer. Some scholars have proposed that there are coarse-grained models allowing simulations of long polymer chains [The molecular weight Mr of MVQ is: (4.0~6.0) × 10r chains ,31. In oTo simulate the molecule evolution at room temperature, the field was set to 298 K. Moreover, the model didn’t take other fillers or impurities into account. Based on the Materials Studio software the Forcite/COMPASS force field was used to conduct optimizations of molecular geometry and energy minimization. The COMPASS force field incorporates many ab initio calculations of quantum mechanics and has many parameters for metals and metal oxides. Moreover, it is the first high quality molecular force field that unites organic and inorganic molecular parameters in the same force field. Its correctness has been verified by a total of 28 molecules of single molecules, liquid molecules and crystal molecules . The 20,As can be seen from Considering research objects contains C, H, O, Si atoms, the Dreiding force field was selected to calculate the structure and properties of molecular aggregates, so MD calculations were carried out at different electric field intensity by the GULP/Dreiding package. The calculations were run for 5000 steps along with a 1 fs time step on the well at a temperature of 298 K. The relaxation time is 1 ps. The electric field was set in the X direction from 0.001 V/Å to 3 V/Å with an interval of 0.2 V/Å and two additional fields 0.01 V/Å and 0.1 V/Å.3 as well as -CH=CH2. As the electric field intensity increases, there is a significantly shrinked molecule with a more obvious helical structure. Meanwhile, group, rotations are larger and the number of -Si-Si- bonds constantly changes. There is a tendency for -Si-O-Si- bridging bonds to form a ring structure with shortened -C-H bonds and the molecular length is significantly reduced.By statistical analysis, obtained trends of total energy and potential energy in each run are shown in At the same time, as can be seen from The potential energy is a part of total energy, and its change trend is basically the same as total energy , which iIt can be found in The molecular structure has undergone a significant degradation with greatly reduced potential energy under further increased intensity. There was no doubt that the molecules’ ability to resist an external electric field is greatly weakened, the internal temperature swinging increases, leading to further destruction of molecular potential energy.Because the molecular chain shrinks helically, making two Si atoms on the -Si-O-Si- bond close to each other, so Si-Si bonds are generated by interatomic van der Waals force and their own changed energy. An unequal number of -Si-Si- bonds which present a scatter point distribution at various electric field intensity is shown in To explore its distribution pattern, a curve fitting Under a constant electric field, the initial reaction of silicone rubber molecule is the atoms of methyl and ethylene groups vibrate violently, chemical bonds are shortened, and neighboring groups are close to each other. During the process, an unequal number of -Si-Si- bonds are generated, which promotes spiraling shrinkage of the molecule. Then the middle protruding of molecular chain presents an inverted U shape.(2)Higher electric field intensity will cause a significant decrease in the total energy of the silicone rubber molecule, especially destruction of molecular potential energy, which results in the degradation of molecular structure. The continuous high electric field intensity will reduce molecular energies and provide a basis for further degradation of the materials.(3)As the electric field intensity increases, the elastic modulus of molecular chain gradually increases. It shows that strong electric field will increase the rigidity of the materials, and the brittleness will become stronger, which will reduce mechanical properties of the materials and accelerate its aging process.Based on molecular simulations, the effects of electric field on the microstructure evolution of silicone rubber molecule were studied. The results can establish theoretical basis for the relationship between the micro morphology and macroscopic characteristics of the materials after further aging and degradation. Furthermore, it can provide references for optimizations of base materials used for composite insulators. What results have been found are as follows:


Saturated fatty acids (SFA) have been reported to promote inflammation. Nevertheless, evidence linking dietary SFA and low-grade inflammation in adolescents is scarce and inconsistent. The modulatory role of physical activity (PA) on fat metabolism and inflammation may provide a potential explanation. Thus, we assessed the association of dietary SFA with high-sensitivity C-reactive protein (hsCRP), a marker of low-grade inflammation, in 15-year-olds, and evaluated possible interactions between dietary SFA and different levels of PA.N = 824). SFA intake was estimated by means of a food frequency questionnaire and PA recorded by accelerometers. Average daily minutes of PA were classified into “sedentary”, “light” and “moderate-to-vigorous” (MVPA), using Freedson’s cut-offs. HsCRP concentrations were measured in serum and categorized into 3 sex-specific levels (below detection limit (I), above 75th percentile (III), in between (II)). Sex-stratified cross-sectional associations between SFA and hsCRP were assessed using multinomial logistic regression, adjusting for potential confounders. Interaction terms were included between SFA and the different PA levels; and if significant interactions were observed, analyses stratified by tertiles of the relevant PA levels were performed. Relative risk ratios (RRR) and 95% confidence intervals (95%CI) were presented for a 1% increase in SFA.Children participating in the 15-year follow-ups of the GINIplus and LISA German birth cohort studies were included (p = 0.008), whereas no significant association was observed in females. A significant interaction was observed with “sedentary” and “light” PA but not with MVPA in both sexes (p < 0.05). Stratified analyses indicated a significant inverse association between SFA and medium hsCRP levels in males in the highest light PA tertile .An inverse association was observed between SFA intake and hsCRP (II vs. I) in males  contains supplementary material, which is available to authorized users. Chronic, low-grade inflammation precedes the onset of cardiovascular diseases , which cIt has been proposed that the interaction with other lifestyle factors might be relevant in determining whether (or to what extent) dietary SFA contributes to chronic low-grade inflammation . PA is fn = 5991) was recruited in Munich and Wesel between 1995 and 1998, and consists of two study arms: an observation arm and an intervention arm. New-borns with a family history of allergy were invited for the intervention arm, in which children were randomised to receive one of three hydrolysed formulas or cow’s milk. The aim was to compare the effect of the different formulae vs. cow’s milk on allergy development in a double-blind controlled trial. Participants with a negative family history of allergy, and those who declined to take part in the intervention trial, were included in the observation arm. The LISA cohort  is a true population-based cohort, recruited in Munich, Wesel, Leipzig and Bad Honnef, between 1997 and 1999. In both studies, information on selected exposures and health outcomes were obtained by means of questionnaires and medical examinations carried out at various follow-up assessments. Exposures and outcomes relevant to the present analyses are described in detail below. Both cohort studies have been approved by their local ethics committees  and written consent was obtained from all participants’ families.The present study used data from the 15-year follow-up assessments of the GINIplus  and LISA (Influence of Life-style related factors on the development of the Immune System and Allergies in East and West Germany) birth cohort studies. Details on the cohorts’ recruitment and follow-up strategies have been described previously and can be found elsewhere , 53. BriHabitual dietary intake was assessed at the 15-year follow-up using a self-administered food frequency questionnaire (FFQ). The applied FFQ was designed at the 10-year follow-up for the estimation of food and nutrient intake in school-aged children over the past year. A detailed description of the FFQ development and its validation can be found elsewhere . BrieflyPA was measured at age 15 years using triaxial accelerometers , worn on the dominant hip for seven consecutive days. The accelerometer has been validated for use in adolescents , and it Serum concentrations of hsCRP were measured in samples collected during the 15-year follow-up medical examinations, using the Roche  Tina-quant CRP (latex) high-sensitive assay, according to manufacturer instructions. Measured hsCRP concentrations were highly skewed, with many observations below detection limit (hsCRP 0.016 mg/dl). Given this non-normal distribution, data categorisation was required for analyses. The variable was hence categorised into three levels separately for girls and boys, considering all children with available hsCRP measurements: all adolescents with hsCRP levels below the detection limit (hsCRP < 0.016 mg/dl) were grouped in the lowest category (I); amongst those remaining, sex-specific 75th percentiles were determined , and those below the 75th percentile, i.e. girls with hsCRP < 0.085 and boys with hsCRP < 0.092 mg/dl, were assigned to the middle category (II); those above the 75th percentile, i.e. girls with hsCRP ⩾0.085 and boys with hsCRP ⩾0.092 mg/dl, were assigned to the upper category (III). While hsCRP values between 0.3 and 1 mg/dl are usually deemed indicative of elevated low-grade inflammation and of higher risk of CVD in adults , these l2 calculated from height and weight measurements obtained during physical examination, unless unavailable (n = 65), in which case obtained from the 15-year follow-up questionnaire), and total daily energy intake .Participants from the 15-year follow-up of GINIplus and LISA studies, with complete data on SFA intake, accelerometer-measured physical activity, and hsCRP, were included in the statistical analyses. Since PA was measured only in Munich and Wesel, the study sample was limited to participants from these two study centres. Sex-stratified associations between SFA intake and hsCRP were assessed using multinomial logistic regression, adjusting for potential confounders: study , region , parental education , pubertal stage , which has been validated in adolescents and includes ratings on body hair growth, voice change and facial hair growth for boys, and body hair growth, breast development and menarche for girls , 68. Forhttps://www.R-project.org/) [In three additional models, separate adjustment for the different PA levels  was carried out. Interaction terms were then included between SFA and the different PA levels. Where a significant interaction was observed, additional analyses were performed, stratified by tertiles of the relevant PA level. Here, we corrected for multiple testing using Bonferroni correction: the α-level was divided by three . This yielded a corrected two-sided α-level of 0.017 (0.05/3 = 0.017). Relative risk ratios (RRR) and 95% confidence intervals (95%CI) are presented for a 1% increase in SFA. The RRR indicates the risk of being in one of the comparison groups relative to the risk of being in the reference group given a 1% change in SFA. All analyses were conducted using R, version 3.3.2 (ct.org/) . Multinoct.org/) .A total of 824 participants  were included in the analyses , which remained significant following further adjustment for PA variables. A dose-response relationship was however not indicated, as this association was observed at the middle hsCRP (II) level but not the upper (III) level.Results from the multinomial logistic regression assessing the association between SFA and hsCRP, including individual adjustment for different PA levels, “sedentary”, “light” and “MVPA”, and their interactions, are displayed in Table p = 0.003).While adjustment for PA levels did not influence the association between SFA and hsCRP, significant interactions with time spent “sedentary” and in “light” PA were observed in females and males. Further analyses were hence carried out stratified by both these PA levels. Results from the stratified analyses are displayed in Fig. The present study assessed the association between dietary SFA and low-grade inflammation, measured by the inflammatory marker hsCRP, in 15-year-old German adolescents. Two aspects stand out among the analyses findings: 1) dietary SFA appears to have no association with hsCRP levels in adolescent females and to be inversely associated with hsCRP in males, albeit only for the middle hsCRP level with respect to the lowest, thus showing no clear exposure-response relationship; 2) a significant interaction between SFA intake and “light” PA likely plays a relevant role, given that the inverse association observed in males was only present among those spending the most time in “light” PA, as indicated by stratified analyses. Although average daily minutes of MVPA were comparable to reports in other adolescent populations , 72, it n = 219), reported no association between dietary SFA and hsCRP [The results of our analyses in females are in line with a number of studies in adults, in which no significant association has been observed between dietary SFA and hsCRP , 36, 75.nd hsCRP . On the nd hsCRP ; while and hsCRP . The incnd hsCRP . Intakesnd hsCRP . Compareintake and status. While the assessment of SFA intake considers the amount of the nutrient consumed, SFA status likely reflects circulating SFA following additional processes such as digestion, absorption, uptake into tissues, and metabolism [It is also possible that contradicting results in terms of SFA may reflect true differences between SFA tabolism . In the tabolism , a finditabolism . Reducedtabolism . Indeed,tabolism , and sevtabolism . A previtabolism . We thertabolism .p < 0.05). It is possible that due to the smaller sample size and greater variance in the highest hsCRP category, there was insufficient power to detect a significant association. Why the inverse association was only observed in males is unclear, but it is possible that sex-specific physiological factors might play a significant role, leading to differences in fat metabolism and the resulting inflammatory profile. For example, testosterone has been shown to enhance lipid oxidation whereas oestrogen enhances fat storage [The inverse association between SFA and the middle category of hsCRP, apparent in male adolescents of the current study, was driven mainly by a specific subgroup with high levels of daily “light” PA. To our knowledge, this is the first study to evaluate the interaction of PA in the association of SFA and low-grade inflammation in adolescents, and hence comparison with other studies is limited. Anti-inflammatory effects of habitual PA in children have been observed , althoug storage , aspectsThe present study benefits from a large, homogeneous study sample, and adds to the limited literature on the association between dietary SFA and low-grade inflammation in adolescents, in a time of heightened discussion concerning SFA and cardiovascular health. Our study includes data from over 800 individuals, greatly exceeding the size of the few observational studies carried out thus far. Our analyses also include the assessment of different levels of accelerometer-measured physical activity, a method not often available in large cohort studies. Accelerometers were worn by participants on the hip, reported to be the best single location to record data for activity detection . FurtherA main limitation when assessing dietary intake is the reliance on subjective measures, which are prone to reporting bias. In the present study the FFQ used to measure dietary SFA was designed to estimate fatty acids and antioxidants in school-aged children . Given tFrom the present analyses, it can be concluded that a higher SFA intake during adolescence, within the ranges observed in the current study, is not detrimental in terms of inflammatory processes in adolescents; although we highlight that this may well depend on the nutrient it replaces. Furthermore, the inflammatory role of SFA might be modulated by the amount of daily “light” PA. Adolescent males with higher SFA intakes, who also participated in greater amounts of “light” PA, presented lower levels of hsCRP than their less active peers. We propose that when evaluating the role of SFA in chronic inflammation, it is essential to differentiate between findings involving SFA status (in serum or plasma) and those assessing dietary SFA, as the latter is likely influenced by important modifiable factors such as PA, which may determine whether an inflammatory response arises.Additional file 1:Figure S1. Study Participants. (PDF 188 kb)Additional file 2:Table S1. Associations between SFA and hsCRP categories stratified by tertiles of PA (Sedentary and Light). (PDF 189 kb)


The spin-dependent electron transport in the ferrocene-based molecular junctions, in which the molecules are 1,3-substituted and 1,3′-substituted ethynyl ferrocenes, respectively, is studied by the theoretical simulation with nonequilibrium Green’s function and density functional theory. The calculated results suggest that the substitution position of the terminal ethynyl groups has a great effect on the spin-dependent current-voltage properties and the spin filtering efficiency of the molecular junctions. At the lower bias, high spin filtering efficiency is found in 1,3′-substituted ethynyl ferrocene junction, which suggests that the spin filtering efficiency is also dependent on the bias voltage. The different spin-dependent transport properties for the two molecular junctions originate from their different evolutions of spin-up and spin-down energy levels. The molecular device is thought to be the substitute for the silicon-based microelectronic device for application in future computers. Over the recent twenty years, molecular electronics has been investigated widely by researchers all around the world. Experimental and theoretical explorations of electronic transport properties in molecular devices have made great progress ,2,3,4.Relative to the conventional electronic device, the spintronic device has many advantages, such as nonvolatility, smaller size, and faster data processing speed . Spin inMetallocene is a complex with a transition metal cation sandwiched between cyclopentadienyl (Cp) rings, and it has unpaired spin components and good chemical stability. Considering it as a possible candidate for molecular devices, the molecular junctions based on metallocene were investigated extensively through experiments and theoretical simulations ,29,30,312) is the most common metallocene. The two Cp rings can be in the staggered or eclipsed conformation and rotate with low resistance about the Cp–Fe–Cp axis  V with a step of 0.01 V are calculated, and the results are presented in EF value at zero bias is set to 0, the bias window is .The transmission spectrum can be employed to analyze the above-current voltage characteristics. According to Equation (1), the transmission coefficients within the so-called bias window  eV in model M1 at 0 V, 0.5 V, 1.0 V, and 1.5 V are presented in The formation of peaks on the transmission spectrum is connected with the molecular orbitals. The contribution of molecular orbitals on the transmission can be analyzed by the projected density of states (PDOS). The PDOS is the density of states which is projected onto some specific atoms of the model. The spin-up and spin-down PDOS curves in two models at the biases of 0.0 V, 0.5 V, 1.0 V, and 1.5 V are plotted in EF has been set to 0 eV, therefore, EF is closer to the HOMO than to the LUMO. It indicates that the HOMO level is the decisive contributor of electric conductance in model M1. A similar situation is also found in model M2. The evolution of HOMO levels of models M1 and M2 is shown in To further understand the spin-dependent transmission characteristics, the molecular projected self-consistent Hamiltonian (MPSH) of the models is investigated. In our work, the Hamiltonian is projected onto the molecule in the middle of the model. For model M1 at zero bias voltage, the highest occupied molecular orbital (HOMO) levels are about −0.12 eV for spin-up state and −0.17 eV for spin-down state, and the lowest unoccupied molecular orbital (LUMO) levels are about 2.03 eV for spin-up state and 2.01 eV for spin-down state. These MPSH energy levels are in accordance with the positions of transmission peaks and PDOS peaks at zero bias voltage. Thus, the transmission peak in the bias window in model M1 originates from the HOMO level. Since Ispin-up > Ispin-down.In order to analyze the high SFE value at the lower bias in model M2, the spin-up and spin-down transmission spectra at 0.05 V are compared, as shown in Itot under a bias voltage is given by Itot = Ispin-up + Ispin-down. According to this relation, it can be found from 2 is important for the molecular conductance. Comparing to the Fe atom in model M1, the one in model M2 may make a more contribution to the electron tunneling from one electrode to the other electrode passing through the molecule, and the conductivity is enhanced accordingly. Therefore, this suggests that the participation of Fe atom should be a more intrinsic reason for the molecular conductance. The characteristic is similar to that in our previous work [The total current ous work .In this work, the spin-dependent transport properties of the molecular junctions based on ferrocene with terminal ethynyl groups are investigated by theoretical simulation with the NEGF−DFT method. For the two models M1 and M2, which are constructed from 1,3-substituted and 1,3′-substituted ethynyl ferrocenes, respectively, our calculated results show that their currents have spin-polarized characteristics. For model M1, the spin-up currents are greater than the corresponding spin-down ones, but the opposite is true for model M2. The SFE values of model M1 keep constant approximately in a broad bias range, and those of model M2 are high at the lower bias. The substitution position of the ethynyl group has a great effect on the spin-dependent transport properties and SFE. The results suggest that ethynyl ferrocene may be treated as a candidate for designing a stable and high-performance spin filtering device.


The samples were characterized by X-ray powder diffraction (XRD), inductively coupled plasma optical emission spectrometry (ICP-OES), N2 adsorption/desorption, and scanning electron microscopy (SEM). It was shown that Cu(II) can be completely reduced to Cu(I), and the monolayer dispersion threshold of CuCl on AC support is 4 mmol·g−1 AC. The adsorption isotherms of CO, CO2, CH4, and N2 on CuCl/AC adsorbents were measured by the volumetric method, and the CO/CO2, CO/CH4, and CO/N2 selectivities of the adsorbents were predicted using ideal adsorbed solution theory (IAST). The obtained adsorbent displayed a high CO adsorption capacity, high CO/N2, CO/CH4, and CO/CO2 selectivities, excellent ad/desorption cycle performance, rapid adsorption rate, and appropriate isosteric heat of adsorption, which made it a promising adsorbent for CO separation and purification.CO is a toxic gas discharged as a byproduct in tail gases from different industrial flue gases, which needs to be taken care of urgently. In this study, a CuCl/AC adsorbent was made by a facile route of physically mixing CuCl CO is toxic to humans because it combines with hemoglobin in the blood to form carboxy-hemoglobin hindering the transportation and release of oxygen in the blood, which leads to death [With the rapid development of C−1 chemistry recently in the chemical industry, carbon monoxide (CO) as a significant resource has been widely applied to prepare a large variety of chemical products, such as formic acid, acetic acid, oxalic acid ester, carbonic acid two methyl ester, anhydride, etc. . Most ofto death . Moreoveto death ,9. Thus,2 or CO. However, it is difficult to control the reduction degree, and Cu(II) is easily over reduced to Cu. In the second process, Cu(I) adsorbents are prepared by direct dispersion and impregnation of CuCl. Hirai et al. [2. Xie et al. [2 and Cu(HCOO)2 as starting materials by the traditional impregnation method followed by activating at the temperature of 583 K [Among the proven technologies for CO separation and purification, adsorption processes, such as pressure swing adsorption (PSA) and temperature swing adsorption (TSA) have the advantages of convenient operation, low energy consumption, low operating cost, etc., and have been widely used in CO separation ,12,13,14s forces ,24,25,26 systems . Two app systems ,25,27, ai et al. ,29 and Ti et al.  obtainede et al.  preparedof 583 K .2 and Cu(HCOO)2 as the initial material. Then, X-ray powder diffraction (XRD), inductively coupled plasma optical emission spectrometry (ICP-OES), N2 adsorption/desorption and scanning electron microscopy (SEM) were employed to characterize the samples. Pure component CO, CO2, N2, and CH4 adsorption isotherms on the adsorbents were measured in a volumetric method. The CO/CO2, CO/CH4, and CO/N2 selectivities of the adsorbents were predicted by using ideal adsorbed solution theory (IAST). The adsorption isotherms were fitted with the Langmuir–Freundlich model, and the corresponding heats of adsorption were calculated. The cyclic CO adsorption on adsorbent was performed to evaluate its repeated availability during the adsorption and desorption cycles. Furthermore, the CO adsorption rate on adsorbent was discussed and reported.Herein, the purpose of this work is to develop CO adsorbent using a solid-state auto reduction–dispersion method with CuCl2·4H2O, 98%) and cupric chloride dihydrate  were purchased from Alfa Aesar Chemical Co. Ltd.. Activated carbon (AC) was purchased from Chengde Jingda Activated Carbon Manufacturing Co. Ltd. .Copper formate tetrahydrate (Cu(HCOO)2 and Cu(HCOO)2 powder to obtain CuCl/AC adsorbent precursors. Then, the obtained precursors were dried at 373 K and activated in a tube furnace at 533 K for 4 h under vacuum. The obtained precursors and CuCl/AC adsorbents were marked as Cu(II)-x/AC and Cu(I)-x/AC , in which the loading of copper is 2, 3, 4, 5, and 6 mmol·g−1 AC, respectively. The as-synthesized CuCl/AC adsorbents were stored in vacuum dry storage in a desiccator.CuCl/AC adsorbents were synthesized following two steps. First, the AC was physically mixed with CuCl2 adsorption/desorption isotherms measured on a surface area and pore size analyzer  after activating the samples at 393 K for 4 h under vacuum. The specific surface areas (SBET) were determined using the BET (Brunauer–Emmett–Teller) method under relative pressure in the range of 0.01 to 0.20. The adsorbed amount of N2 at p/p0 = 0.98 was employed to calculate the total pore volume . Scanning electron microscope  was used to observe the samples’ morphology. Cu contents were measured by inductively coupled plasma optical emission spectrometry .Powder X-ray diffraction (XRD) patterns of the samples were recorded by a Shimadzu LabX XRD-6000 system  in the 2θ range of 5 to 35° using CuKα1 (λ=1.54056 Å) radiation operated at 40 kV and 30 mA. The pore volume and surface area of the samples were calculated from N2, CH4, and N2 adsorption isotherms were measured at the temperature required using a static volumetric apparatus . During the adsorption measurements, the temperature was maintained by circulating ethanediol-water from a bath with setting temperature. The adsorption capacity was determined from the adsorption isotherm measured at 298 K. Ultrahigh purity grade CO (99.99%), CO2 (99.999%), CH4 (99.99%), and N2 (99.999%) were used without any purification.Before the adsorption measurements, the samples were degassed under vacuum while heating up to 393 K for 4 h. The CO, CO2 and CuCl2 [2 and Cu(HCOO)2 disappeared, and the Cu(I)-5/AC sample displayed only a relatively weak peak (2θ = 28.5°) of CuCl [2 and CuCl2 were transformed into CuCl after activation. Meanwhile, the absence of CuCl diffraction peak on Cu(I)-2/AC, Cu(I)-3/AC, and Cu(I)-4/AC samples might be due to the well dispersion of CuCl on the AC surface beyond the detection limit of XRD [−1, the appearance of the characteristic peak of CuCl implies that the crystal size of CuCl on the AC surface increased with the increase of CuCl loading, which was able to be detected by XRD with the CuCl loading higher than 5 mmol·g−1.The XRD patterns of the copper loaded AC samples before and after activation are presented in nd CuCl2 ,34 can b of CuCl , suggestt of XRD . By furt2 adsorption/desorption isotherms of the Cu(I)-x/AC samples at 77 K. The N2 adsorption gradually decreased with increasing CuCl loading. Total) and BET surface area (SBET) gradually decreased with the increase of CuCl loading, indicating that CuCl had been loaded into the pores of the parent AC. As the CuCl loading increased, more and more surface within the pores were occupied by CuCl, which may result in a further decrease of VTotal and SBET. As CuCl was well dispersed on the surface of AC when the CuCl loading was below 4 mmol/g, the average pore sizes decreased with the increase of CuCl loading. Smaller pores were filled with the further increase of the CuCl loading. Therefore, the increased average pore size resulted from the percentage increase of the available larger pores in AC, which is similar to the observation by Ramli et al. [i et al. .−1. The maximum value of adsorption capacity was 45.4 cm3·g−1. With the continuous increase of the CuCl loading, the CO adsorption capacity of CuCl/AC with the copper loading of 5 mmol·g−1 was almost the same as that with 4 mmol·g−1. The decrease of CO adsorption occurred with further increasing the copper loading to 6 mmol·g−1. This phenomenon can be ascribed to the following reason. The more copper loaded, the more active sites of CuCl/AC adsorbents present, which would enhance CO adsorption. However, the increase of copper loading also resulted in a decrease of surface area for CuCl/AC adsorbents, as shown in −1. When the copper loading reached 6 mmol·g−1, on the one hand, the amount of adsorbed CO decreased because the decrease of surface area; on the other hand, the Cu(I) started to agglomerate on AC surface with considerable copper loading, resulting in the low utilization of active sites.It can be observed from In addition, 2, CH4, and N2 on Cu(I)-4/AC in the pressure range of 0 to 100 kPa. The adsorption of CO2, CH4, and N2 on Cu(I)-4/AC increased almost linearly with pressure, while the adsorption isotherm of CO on Cu(I)-4/AC presented a type-Ι isotherm [2/CH4/N2 mixed gas.isotherm , that is2, CO/CH4, and CO/N2 selectivities with the equimolar CO/CO2, CO/CH4, and CO/N2 mixture. The L-F model can be expressed asThe Langmuir–Freundlich (L-F) model and IAST were employed together to calculate the CO/CO2, CH4, and N2 were fitted by the L-F model [2, CO/CH4, and CO/N2 selectivities were predicted by IAST theory [2, CO/CH4, and CO/N2 selectivities decrease gradually with increasing pressure. Nevertheless, the CO/CO2, CO/CH4, and CO/N2 selectivities on Cu(I)-4/AC were still up to 2.6, 8.0, and 34.3 at 100 kPa, respectively, which suggests that it has the potential for the effective separation of CO from the gas mixtures. 2 selectivity among the selected adsorbents.First, the adsorption isotherm of pure CO, CO-F model . After tT theory ,40. Fina2 . −1 for common physical adsorption and >80 kJ·mol−1 for chemical adsorption [−1 in the whole pressure range, suggesting that the strength of complex adsorption is between physisorption and chemisorption. Such isosteric heat is not only propitious to adsorb CO, but also liable to desorb CO with a normal engineering operations .Isosteric heat of adsorption is a significant thermodynamic parameter to characterize the interaction between the adsorbate and the adsorbent and to design a gas adsorption separation process, which can be calculated by Clausius–Clapeyron equation  as[∂lnPsorption . The val2, CH4, and N2. Typically, the requirement of the adsorption process in industrial applications is shorter than 1 min [2, CH4, and N2 in a PSA process.In addition, it is also crucial that the CO adsorption rate need to be quite rapid for potential applications of CuCl/AC adsorbent in the adsorption-driven separation of CO from gas mixtures containing CO, COan 1 min . Here, w2 . As shown in ondition ,45), whi2 and Cu(HCOO)2 as the initial material by a solid-state auto dispersion method. CuCl2 and Cu(HCOO)2 can be transformed into highly dispersed CuCl with activation at 533 K under vacuum atmosphere. The CO adsorption capacity increased with transformed CuCl loading until 4 mmol·g−1 and then decreased afterward. The CO adsorption capacity of Cu(I)-4/AC achieved 45.4 cm3·g−1, and the CO/CO2, CO/CH4, and CO/N2 selectivities were up to 2.6, 8.0, and 34.3 at 100 kPa, respectively. In addition, the isosteric heat of adsorption on Cu(I)-4/AC was about 50 kJ·mol−1. The CO adsorption capacity almost remains constant during six times cyclical adsorption and rapid adsorption kinetics at the adsorption process. Those excellent properties of Cu(I)-4/AC adsorbent would make it a promising adsorbent for CO separation and purification.CuCl/AC adsorbents for the separation of CO have been successfully obtained using CuCl


Gracilaria lemaneiformis sulfated polysaccharide  was degraded by H2O2 to obtain seven degraded fragments, namely, GLP1, GLP2, GLP3, GLP4, GLP5, GLP6, and GLP7, with molecular weights of 106, 49.6, 10.5, 6.14, 5.06, 3.71, and 2.42 kDa, respectively. FT-IR and NMR results indicated that H2O2 degradation does not change the structure of GLP polysaccharides, whereas the content of the characteristic −OSO3H group (13.46% ± 0.10%) slightly increased than that of the natural polysaccharide (13.07%) after degradation. The repair effects of the polysaccharide fractions on oxalate-induced damaged human kidney proximal tubular epithelial cells (HK-2) were compared. When 60 μg/mL of each polysaccharide was used to repair the damaged HK-2 cells, cell viability increased and the cell morphology was restored, as determined by HE staining. The amount of lactate dehydrogenase released decreased from 16.64% in the injured group to 7.55%–13.87% in the repair groups. The SOD activity increased, and the amount of MDA released decreased. Moreover, the mitochondrial membrane potential evidently increased. All polysaccharide fractions inhibited S phase arrest through the decreased percentage of cells in the S phase and the increased percentage of cells in the G2/M phase. These results reveal that all GLP fractions exhibited repair effect on oxalate-induced damaged HK-2 cells. The repair ability is closely correlated with the molecular weight of the fractions. GLP2 with molecular weight of about 49.6 kDa exhibited the strongest repair effect, and GLP with higher or lower molecular weight than 49.6 kDa showed decreased repair ability. Our results can provide references for inhibiting the formation of kidney stones and developing original anti-stone polysaccharide drugs.Natural  Sargassum fulvellum polysaccharide to obtain three polysaccharide fractions with low molecular weights of 2, 23, and 36 kDa; the antioxidant and anticoagulant activities increased with decreasing molecular weight of the polysaccharides. Zhu et al. . In the present study, the monosaccharide composition and type of sugar residues of GLP were analyzed by 1H NMR, 13C NMR, and FT-IR spectroscopy methods. The signals at δ 4.43 and 5.34 ppm in the 1H NMR spectra correspond to the H1 of β-D-galactose and 3,6-α-L-galactoptoside, respectively. The signals at δ 102.35 and 99.48 ppm in the 13C NMR spectra correspond to the anomeric protons of β-D-galactose and 3,6-α-L-galactoptoside, respectively. The peak at 1370 cm−1 in the FT-IR spectra is assigned to ester sulfate, and the peak at 1260 cm−1 corresponds to the stretching vibration of S=O [β-D-galactose and 6-O-sulfate-3,6-L-galactose  free radicals produced by H2O2 during degradation will break the highly compact sugar chain structure of natural polysaccharide, thereby exposing the −OSO3H and −COOH groups of the polysaccharide, and (2) the water solubility of the degraded polysaccharide slightly increased and was higher than that of nondegraded GLP0 because of the large molecular weight of the latter, resulting in the concealment of small parts of the acidic groups of polysaccharides [As shown in Figures charides , 39.2O2 degradation of the crude Monostroma latissimum polysaccharide; the −OSO3H group content  increased with decreasing molecular weight of the polysaccharide. Sun et al. [2O2 and Vc to degrade Pavlova viridis and Sarcinochrysis marina Geitler. When the molecular weight of P. viridis decreased from 3645 kDa to 55.0 kDa, the −COOH content increased from 3.41% to 8.78%. The −COOH content increased from 5.82% to 9.99% when the molecular weight of S. marina Geitler decreased from 2595 kDa to 169 kDa.Similar results were reported by previous studies , 39. Zhan et al.  used H2Oμg/mL GLPs, and the MDA content was obviously reduced compared with the injured cells. In addition, the present results indicate that cell viability decreased  and MDA  and damaecreased  and the ecreased  when HK-ecreased . Subha arphology .2+ influx and depolarization of the mitochondria [ψm  increased Bcl-2 protein expression in the mitochondria; the polysaccharide inhibited Bax translocation, cytochrome c release, and caspase activation, resulting in an increase in Δψm of the cell. Sparassis crispa polysaccharide fraction with a molecular weight of 75 kDa reduced the accumulation of reactive oxygen species, blocked Ca2+ influx, and prevented depolarization of the mitochondrial membrane potential to protect PC12 cells against L-Glu-induced injury [3H and −COOH functional groups are rich in GLP, which can scavenge reactive oxygen species [Mitochondrial membrane potential (ΔΨm) is higher in normal cells than that in damaged cells . When thchondria , resultidria [ψm . GLP fra Figures ; that isi et al.  reportedd injury . GLP can species .In damaged cells, the percentage of cells in the S phase (51.9%) increased but that in the G2/M phase decreased . The celThe high molecular weight of polysaccharides limited their physical properties, such as highly compact molecular structure, large molecular size, and low water solubility, resulting in decreased possibility of migrating to the cell membrane ; the polPorphyridium cruentum and obtained six degraded polysaccharide fractions, with molecular weights of 6.53, 256, 606, 802.6, 903.3, and 1002 kDa; the polysaccharide fraction with molecular weight of 6.53 kDa exhibited the optimal effect on enhancing immunomodulatory ability.For example, the biological availability of low-molecular-weight heparin is higher than that of the ordinary heparin, the former thereby inhibiting atherosclerosis . A previG. scabrabunge, is lower than that of GSP-3, with a high molecular weight of 58 kDa.If the polysaccharide molecular weight is too low, it cannot form an active polymer structure for the biological activity. Liao et al.  studied Polysaccharides with appropriate molecular weight have high freedom degree and small steric hindrance; they require less energy to diffuse into cell membrane breach ; therefoThe repair effect of GLP on damaged HK-2 cells could be due to polysaccharide molecules, which are dispersed to the damaged cell membrane gap to repair the cell or adhered to the cell membrane to block further oxidative damage by oxalate; the dispersion and adhesion of the polysaccharide are closely correlated with its molecular weight . The hiRenal tubular cell injury plays a key role in the pathophysiological processes of renal stone diseases. Abnormally elevated urinary oxalate induces tubular dysfunction or damage, thereby promoting the retention of CaOx crystals. This phenomenon is believed to be a prerequisite for the eventual formation of kidney stones . Our res2O2 concentration during degradation of crude GLP. GLP consists of β-D-galactose and 6-O-sulfate-3,6-α-L-galactose. H2O2 degradation does not change the structure of GLP polysaccharides, whereas the contents of the −OSO3H and −COOH groups of the polysaccharides slightly increased after degradation. Various polysaccharide fractions can repair oxalate-induced oxidatively damaged HK-2 cells. After being repaired by polysaccharides, the cell viability and SOD activity increased, the amount of LDH and MDA released decreased, the cell morphology was gradually restored to normal cells, the mitochondrial membrane potential increased, the percentage of cells in the S phase decreased, and the percentage of cells in the G2/M phase increased. The repair ability of each polysaccharide fraction of GLP is associated with its molecular weight. GLP2 with a molecular weight of 49.6 kDa exhibited a superior repair effect than did other degraded polysaccharide segments and crude polysaccharide. Moreover, polysaccharides with a molecular weight largely deviating from 49.6 kDa elicited weak repair capacity. Our results suggested that degraded GLP fractions, especially GLP2, could be developed into novel anti-stone polysaccharide drugs.Seven fractions with molecular weights of 106, 49.6, 10.5, 6.14, 5.06, 3.71, and 2.42 kDa were obtained by controlling H


Cichorium intybus L.) is a traditional European crop that is highly appreciated for its contents of bioactive compounds, especially phenolics, which have high antioxidant activities. Among other factors, agricultural practice might affect the contents of these bioactive compounds, which are also important from a nutritional point of view, and affect the shelf-life.Chicory  was evaluated using Folin–Ciocalteu reagent, and total flavonoids content (TFC) using the aluminium chloride method. Four different chicory cultivars were included: ‘Treviso’, ‘Verona’ and ‘Anivip’ as red cultivars; and ‘Castelfranco’ as a red-spotted cultivar. These were grown in pots under controlled glasshouse conditions using organic and/or mineral fertilisers. The combination of organic and mineral fertilisers during red chicory growth resulted in significantly higher in-vitro and in-vivo AOPs compared to the control. For the red-spotted cultivar ‘Castelfranco’, this combined organic and mineral fertilisation decreased AOPs in vitro and increased AOPs in vivo. Among the cultivars examined, ‘Castelfranco’ treated with combined organic plus mineral fertilisers showed the highest AOP in vivo, accompanied by the lowest TPC and TFC.The antioxidant potential (AOP) of chicory plants treated with different fertilisers was investigated in vitro using DPPH radical scavenging and in vivo using the yeast These data show that application of different fertilisers has different impacts on red and red-spotted chicory cultivars in terms of TFC and TPC, which for red-spotted chicory resulted in different AOPs in vitro and in vivo. The in-vitro AOP is well reflected in the in-vivo AOP for the red chicory cultivars, but less so for the red-spotted cultivar ‘Castelfranco’. Based on the in-vivo AOPs for these chicory cultivars analysed, the combined organic plus mineral fertiliser treatment is recommended. Cichorium intybus L. is commonly known as chicory. It belongs to the family Asteraceae, and is widely distributed around the world, according to various uses. Chicory plants can be cultivated for food as the leaves, rosettes and heads, which are usually eaten raw in salads. Chicory has become an important vegetable and technical crop in many temperate regions over the last decade, especially in Europe, Asia and North America [ America .One of the core ideas behind organic production compared to conventional production is that the cropping system should be less dependent on the import of resources, and its negative effects on the surrounding environment should be minimised . OrganicA number of comparative studies have shown higher levels of phenolic compounds (phenolics) in organic plant products –8, althoPhenolics are secondary metabolites that have physiological and morphological importance in plants, including chicory. As well as contributing towards the colour and sensory characteristics of vegetables, phenolics have important roles in plant growth and reproduction . ChicoryAnthocyanidins have been reported to reduce intracellular reactive oxygen species . In our A rich area of research has been developed to investigate the effects of bioactive plant components in foods that are not related to direct antioxidant actions. These bioactive plant components should thus be considered simply in terms of their chemical properties, and not automatically related to any equivalent function in vivo. Therefore, it is essential to determine the antioxidant effects of such bioactive plant components at multiple levels, both in vitro and in vivo, to obtain the full picture of their activities, which might be relevant to various physiological and pathological states in humans .There is strong epidemiological evidence that antioxidants derived from fruit and vegetables can protect the body against various diseases. Chicory is a good source of phenolics , 15 thatTo the best of our knowledge, there have not been any studies on the in-vivo antioxidant activities of chicory cultivars produced using different fertilisers . The aim of the present study was thus to determine the in-vitro and in-vivo AOPs of four chicory cultivars to define how they are influenced by commercially available organic and/or mineral fertilisers. Further, we wanted to find out how the in-vitro AOP of chicory correlates to the in-vivo AOP, according to cultivar and fertiliser type.The influence of the fertiliser treatments on in-vitro and in-vivo AOP for the four chicory cultivars is shown in Fig. For all four of the chicory cultivars treated with the different fertilisers, TPC and TFC were measured Figs. :Table S3Table The Pearson’s correlation coefficients between in-vivo and in-vitro AOP, TPC and TFC for the individual chicory cultivars are reported in Table A biplot was constructed for the first two functions, which shows how the treated cultivars are different and which parameter is mainly responsible for the variation. The discrimination across the four variables determined in the chicory samples for these three different fertiliser treatments and the Control is shown in Fig. Vegetables, including chicory, are rich external sources of dietary antioxidants that are important for the human body . StudiesFor the red cultivars ‘Treviso’, ‘Verona’ and ‘Anivip’ compared to the Control, the in-vitro AOPs were highest for the Organic+Mineral fertiliser treatment. The exception was the red-spotted cultivar ‘Castelfranco’, which showed highest in-vitro AOP for Organic fertiliser treatment, while the Organic+Mineral fertiliser treatment resulted in the lowest AOP among these fertiliser treatments. Treatment of the red cultivars with other fertilisers  mostly decreased in-vitro AOPs, with the exception of cultivars ‘Treviso’ and ‘Anivip’. Application of different fertilisers for cultivar ‘Castelfranco’ resulted in different effects that depended on the fertiliser type  between in-vitro AOP and TPC, in-vitro AOP and TFC, and TPC and TFC  were included: three red cultivars ‘Treviso’ , ‘Verona’  and ‘Anivip’ , and one red-spotted cultivar ‘Castelfranco’ . The seeds were purchased from commercial seed companies mentioned above. Selected cultivars of chicory were inscribed in the Slovene list of varieties under register numbers: CCI003 ‘Anivip’ [Four commercial cultivars of chicory . Along with the control (no added fertilisers), different types of fertilisers were tested: an organic fertiliser, a mineral fertiliser, and a combination of an organic and a mineral fertiliser. In each of these four cultivars, the same four treatments were applied in a completely randomised factorial design, as: no added fertiliser (Control); addition of the organic fertiliser Plantella Organic ; addition of the mineral fertiliser ENTEC perfect ; and addition of the combination of the organic fertiliser Plantella Organic (2.5 g/pot) and the water-soluble mineral fertiliser Kristalon Blue . Sowing time and method, plant care, water-soluble fertilizer application during the growth and harvesting time and process was carried out as previously described by Sinkovič et al. [For the preparation of samples only undamaged uniform leaves were collected. For each sample, 10 g fresh leaves were chopped up using a ceramic knife, and extracted in a plastic vial with 10 g 100% methanol. The tissue in the methanol was then homogenised using a laboratory mixer . The samples were stored at − 20 °C until analysed.To determine the dry matter content (%DM), leaves from all of the plants were dried in a laboratory oven at 80 °C for 28 h. The dry matter content ranged from 6.8 to 14.8% of the leaf fresh weight (FW).2DCFDA) was from Sigma-Aldrich .The water used for sample extraction and analysis was from a Milli-Q water purification system . Methanol, Folin–Ciocalteu reagent, the standards of Trolox, gallic acid and quercetin, and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) were from Merck . 2′,7′-Dichlorofluorescin diacetate (H2 = 0.9900).The in-vitro AOP was determined using the DPPH free radical scavenging assay . For theg, 5 min) and the supernatant was filtered . The in-vivo AOP was determined in the yeast Saccharomyces cerevisiae from the Culture Collection of Industrial Microorganisms  by measuring intracellular oxidation. The cultivation of S. cerevisiae was carried out in YPD medium  at 28 °C and 220 rpm. In the stationary phase the yeast cells were centrifuged , washed once and resuspended in phosphate-buffered saline to reach 1 × 108 cell/mL. The S. cerevisiae were further incubated at 28 °C for 96 h, with agitation at 220 rpm. They were then treated with the chicory methanol extracts at 1% (v/v) for 2 h at 28 °C, with agitation at 220 rpm. These treatments were sampled for determination of S. cerevisiae intracellular oxidation.For determination of the in-vivo AOP, 2 mL defrosted leaf homogenate was centrifuged , which reacts with oxidants. This was added as H2DCFDA, which penetrates the plasma membrane and is hydrolysed inside the cells by non-specific esterases. The non-fluorescent H2DCF that is produced can then be oxidised to the fluorescent 2`,7`-dichlorofluorescin (DCF), the levels of which were determined fluorimetrically [The trically .S. cerevisiae cells from 2-mL cell cultures were sedimented by centrifugation , and washed three times with 50 mM potassium phosphate buffer (pH 7.8). The cell pellets were then resuspended in 50 mM potassium phosphate buffer at 10% (v/v), and preincubated at 28 °C for 5 min. The reactive oxygen species sensing dye H2DCFDA was added from a 1 mM stock solution in methanol, to a final concentration of 10 μM. After incubation for 20 min at 28 °C with agitation at 220 rpm, with the fluorescence of the cell suspensions was measured using a microplate reader . The excitation and emission wavelengths of DCF were 488 nm and 520 nm [For this assay, the d 520 nm . The dat2CO3 was added. After an incubation for 2 h at 20 °C [The TPC was determined using a spectrophotometer and following the Folin–Ciocalteu method, as first described by Singleton and Rossi , and as at 20 °C , absorpt2 = 0.9981). The data are expressed as mg QE/100 g FW.The TFC was measured according to a method described previously . Before The experimental data were evaluated statistically using SPSS, version 15.0 for Windows, as the evaluation version . Descriptive statistics were calculated for each chicory cultivar and fertiliser treatment. The data were tested for normal distribution; and the main effects of fertiliser treatment , cultivar  and fertiliser treatment × cultivar, which were tested using the general linear model procedure. Means were calculated for the experimental groups using the least-squared means procedure, and were compared at the 5% probability level. Here, the interaction of fertiliser treatment × cultivar did not have any effects on the data, while each factor did have effects individually . The relationships between the parameters observed were examined according to Pearson’s correlation coefficients. Linear discriminant analysis of the data was performed for in-vitro AOP, in-vivo AOP, TPC and TFC, to model the differences among the classes of data regarding the fertiliser treatments. A biplot was constructed for the first two functions, to illustrate how the treated cultivars varied and to define which parameter defined the greatest variation.Additional file 1: Table S1. Influence of the fertiliser treatments on the in-vitro and in-vivo antioxidant potential (AOP) for the four chicory cultivars.Additional file 2: Table S2. Influence of the fertiliser treatments on the total phenolics content for the four chicory cultivars, as determined by the Folin-Ciocalteu method.Additional file 3: Table S3. Influence of fertiliser treatments on the total flavonoids content for the four chicory cultivars.