PDLLA has been studied for the first time as a separator,taking into account its amorphous character that could facilitateelectrolyte incorporation into the polymer matrix and influence ionicconductivity, together with CNCs, for reducing the hydrophobicityof the scaffolds. The embedding of the nanocrystals in the scaffoldswas confirmed by X-ray diffraction analysis and attenuated total reflectanceFourier transform infrared spectroscopy. The polymer combination influencedthe nanofibrous morphology as evaluated by scanning electron microscopyand modulated the electrochemical behavior of the membranes that wasinvestigated through linear sweep voltammetry, cyclic voltammetry,and electrochemical impedance spectroscopy tests. Among the studiedcategories, the P12 series displayed a nonhomogeneous electrolyteresistance and electrochemical stability, differently from P10, whoseresults suggested their application in LIBs with standard formulation,as confirmed by a preliminary performance test of the P10N6 formulationin a full Li-ion cell configuration.This work concerns the study of electrospun scaffoldsas separatorsfor aprotic lithium-ion batteries (LIBs) composed of the amorphouspoly- Current research trendsfocus on enhanced substitutes for safety and cost reasons; nevertheless,unfortunately, many alternative materials settled these aspects tothe detriment of performances.10Typically, LIBs consist of a transition metal-layered oxide andpolypropylene(PP) separators have been widely used due to their high mechanicalstrength and chemical stability. However, their low porosity and electrolyteretaining, as well as poor thermal stability and safety, representedweak points in their applications.For this reason, the mostimportant drivers in the research onseparators deal with the overcoming of polyolefin separator drawbacksand a careful control of the environmental impact of alternativesfrom both the chemical composition and the manufacturing points ofview.co-hexafluoropropylene)(PVDF-HFP) and poly(vinylidene fluoride-co-trifluoroethylene)(PVDF-TrFE)25 afforded promising results. However, separators obtained from naturaland biodegradable sources are gaining attention. Polyacrylonitrile(PAN),27 polyimide (PI),29 and cellulose derivatives30 are justsome examples of the exploring ones.In this sense, electrospinning turned out to be particularlyhelpful.Several polymeric materials have been investigated, both in combinationthereof and with nanofillers as additives, for nonwoven separatordevelopment: polyvinylidene fluoride (PVDF) and its copolymers aspoly is another valuable alternative:it is a biocompatible and biodegradable polyester used in differentfields, ranging from biomedical38 different from theraceme poly-d,l-lactide (PDLLA), with a lower crystallinedegree, the last form being never tested in LIBs. To enhance batteryperformance, oxide ceramics are typically used as fillers to reducethe crystallinity of polymer separators, since this feature facilitatesthe electrolyte incorporation into the polymer matrix and influencesionic conductivity.18However, both the pure enantiomeric forms ofPLA (PDLA and PLLA)are semicrystalline polymers,However, oneof the main weaknesses of polyesters as aprotic batteryseparators is their hydrophobicity analogous to polyolefin. On theother hand, a better control of hydrophilicity/hydrophobicity canbe achieved by embedding ceramic additives within the polymer matrixes,such as cellulose nanocrystals.39In fact, the polar groups ofCNCs interact with the electrolyteenhancing the uptake and resulting in an improved wettability of theseparators. Equally, hydroxyls strongly interact with Li ions, redistributingthem to a homogeneous deposition on the anode and suppressing lithiumdendrites.42Cellulose nanomaterialscan be extracted by exploiting differentsources and obtaining characteristics which depend on the hydrolysisconditions.44 and electrolytes45 to the separators.47In recent years, several studies have beenreported to improveLIB performances in employing cellulosic materials from anodes48 to drug release.49Moreover, besides their use for designing sustainable energystoragedevices, cellulose-based polymer composites are employed in nanofibrousmatrices for applications which range from water filtrationHere, we draw a comprehensive outline of a class of environmentallyfriendly battery separators by employing the amorphous PDLLA as apolymer host with a convenient blend of CNC fillers and investigatehow their combination in different ratios can modulate the electrochemicaland swelling properties, as well as the nanofibrous morphology.3) was obtained from FormFutura; cellulose nanocrystals were bought from CelluForce,and 1,1,1,3,3,3-hexafluoro-2-propanol (HFP) was obtained from IrisBiotech GMBH. Dimethyl carbonate (DMC) and ethylene carbonate (EC)were purchased from Levanchimica SRL. Commercial-grade reagents andsolvents were used without further purification, except where otherwiseindicated.PDLLA . The obtained segmentedimages automatically allowed the determination of the distributionof fiber diameters and porosity with the same software.Gray-scale images weremanually segmented into binary black (background) and white (fiber)images using ImageJ (Java open source) software supplied with theDiameterJ plug-in  was considered statistically significant.Datahave been reported as mean ± standard deviation. Fiberdiameter and porosity were measured by using the one-way analysisof variance method and Tukey’s test. A 6 EC:DMC 1:1 V/V were performed in a glovebox, after that themembranes were dried in a buchi oven. The membranes were cut intoseparators of 18 mm diameter and immersed in the electrolyte overnight.Five membranes of each sample were used for the analysis.The swelling tests in 1 M LiPFWw and Wd are the masses (g) of the wetand dry scaffold, respectively.Thesolution and the electrolyte absorbed by each scaffold were calculatedaccording to the following equation 1:1Ww andV = 10mV in the 100 kHz–1 Hz frequency range in the Li/Li configurationcell. Performance tests in full lithium-ion cells have been carriedout by galvanostatic cycling at 30 °C at various current rates.Cells have been assembled by coupling a lithium–nickel–cobalt–aluminumlayered oxide (NCA) commercial positive composite electrode  with a commercialnegative composite electrode . The P10N6 separator has been used(see below) and swelled using a 1 M dimethyl carbonate/ethylene carbonatecommercial solution  as the electrolyte.Galvanostatic cycling tests have been carried out using an MTI Corpbattery analyzer.The electrochemical stabilitywindows were evaluated by linear sweep voltammetry (LSV) for anodicscan and cyclic voltammetry (CV) for cathodic scan. SuperP carboncasted on aluminum (cathodic) or copper (anodic) foil was used asthe working electrode and a lithium foil as the counter electrode.The voltage range used was 0–6 V for LSV and 0.01–2.5V for CV, while the scan rate was 1 mV/s. Electrochemical impedancespectroscopy (EIS) tests have been performed using an Ivium Vertexinstrument by applying a sinusoidal Δ51Our work aimed to design ecofriendlyelectrospun battery separators;therefore, the amorphous character of PDLLA for improving batteryperformances, along with its recyclability and biodegradability, isconvenient. Rather than inorganic nanofillers, CNCs have been selectedfor their superior capacity to enhance composite scaffolds’mechanical, chemical, and electrochemical properties.54Therefore, a set of different PDLLA/CNC membranes was developedby electrospinning polymer blends described in Neat PDLLA membranes  wereobtained from solutionspresenting polymer quantities ranging from 8.0 to 12.0% (w/V) in HFP.To each of them, CNCs were added after being sieved in a concentrationranging from 3 to 9% (w/w), obtaining hybrid PDLLA/CNC membranes .55For evaluating CNC’s incorporationinto the hybrid scaffolds, P12 and P12N3–P12N9 hybrid electrospunscaffold diffractograms were recorded as representative behavior ofall samples and compared with the CNC powder as a reference. Thislatter .To confirm further the nanofillerpresence, the hybrid membranes were characterized at a molecular levelby ATR-FTIR and compared with the CNC powder as a reference and to aliphatic C–H stretching vibrations (filled black circle).Typical of carbohydrates is the band at ∼1360 cm–1, which is related to the symmetrical C–H and O–H bendingvibrations (filed red circle) the band at ∼1184 cm–1 associated C–O–C asymmetric stretching vibrations(filled orange circle) of β-(1 → 4) glycosidic bond incellulose and hemicelluloses as well as the three bands at ∼1120,1088, and 1044 cm–1 associated with the stretchingvibrations C–CO stretching vibrations (filled brown circle).58Regarding CNCs (black curve),two broad bands, from ∼3600 to ∼3000 cm–1 the aliphatic C–H and C–H3 stretching vibrations (filled black circle) and at ∼1750cm–1 the ester C=O stretching vibrationsband (filled blue circle). Moreover, in the region from ∼1454to ∼1330 cm–1, the spectrum displayed bandsdue to C–H3 (filled green circle) and C–Hbending vibrations (filled red circle). This latter overlapped at∼1267 cm–1 to ester C–O–C stretchingvibrations,59 which generated bands at∼1181 and at ∼1084 cm–1 (filled browncircle).60P12 (red curve), P12N3 (blue curve), P12N6 (purple curve),andP12N9 (green curve) showed in the region from ∼2998 to ∼2943cm–1  and with polysaccharide C–H bending vibrationsoverlapped with C–O stretching vibrations (filled purple circle),respectively. Moreover, this feature was also confirmed by the increaseof peaks in the region from ∼1200 to 1000 cm–1 where polysaccharide C–C and C–O symmetrical stretchingfall (filled orange and brown circles).61In the region of interest, it can be seen thatthe increase ofbands’ intensity at 1330 and at ∼1267 cm62 Moreover, the porosityshould ideally fall in the 40–60% range to ensure a balancebetween good ionic conductivity and mechanical strength, necessaryfor battery operation.63Morphological characterization of scaffoldswas conducted by SEM image analysis, focusing, besides fiber diameter,on structure uniformity and porosity since they are fundamental aspectsin separator design. Both strongly influence LIBs’ performanceand safety: it is well known that the absence of defects promotesthe electrolyte absorption and ion transport between the electrodesand prevents short circuits in the cell thanks to the good mechanicalstrength of mats.Figure S2a,d) presented beaded randomly oriented fibers, as well as P10N3 . On the contrary, defectlessmorphology has been detected in P12N3 .As seenin 17This feature could be considered a consequenceof increasing viscositydue to higher polymer concentration: a parameter responsible, in addition,for the enlargement of fibers and, consequently, the reduction ofporosity.S3).The average diameterof fibers as a frequency function is representedas histograms 4 and S3.p* ≤ 0.01) of mean diameters  for P12 pure polylactide mats. Nanocrystals’addition to the electrospun mixture produced both enlargements andreductions of fibers’ mean diameter compared with neat PDLLAscaffolds, as reported in Table S1. Thesecontrasting features are respectively associated with an increasein the viscosity and electrical conductivity of the polymer blendscontaining different percentages of CNCs.53Pure polylactide electrospun mats P8, P10, andP12 possessed meandiameters of 402.93 ± 21.56, 410.37 ± 11.30, and 716.10± 18.06 nm, respectively. Statistical analysis displayed a statisticalincrease (p* ≤ 0.01) of the mean diameterin the case of P8N9 (483.83 ± 21.72 nm) in comparison with P8,P8N3, and P8N6, respectively, with 402.93 ± 21.56, 393.60 ±3.65, and 384.73 ± 12.83 nm . For 10% w/V PDLLA, the addition of nanocrystals led to all statisticallydifferent (p* ≤ 0.01) samples: P10N3, P10N6,and P10N9 showed higher values in terms of mean fibers’ diameterwith respect to pure P10 (410.37 ± 11.30 nm), respectively, with746.03 ± 43.19, 617.93 ± 24.55, and 508.97 ± 4.59 nm.However, a statistical reduction has been observed with a growingquantity of nanofillers .The addition of CNCs to 8% w/V PDLLA led to a statisticallydifferentincrement (p* ≤ 0.01) increments andreductions of fiber diameter both in comparison to P12 (716.10 ±18.06 nm) and among composite scaffolds with 686.80 ± 25.12,571.90 ± 17.66, and 898.07 ± 34.42, respectively, for P12N3,P12N6, and P12N9 . These featurescould be ascribed simultaneously to the effect of nanocrystals onviscosity and electric conductivity of polymer blends.12% w/V PDLLA nanocomposite fibers with CNCs reported at the sametime statistical (Table S1): depending on data above reported, P8 and P10 displayed respectivelyvalues of 48.72 ± 3.07 and 53.50 ± 3.89%, with a statisticallyrelevant (p* ≤ 0.01) drop to 37.11 ±1.00 in the case of P12 . Thereby,the addition of CNCs to 8% w/V PDLLA led to a reduction of porosity,moving from 3 to 9% w/w CNC-loading level, despite in P8N3 it wasstatistically higher (67.19 ± 4.10%) than the pure P8 scaffoldand composite ones P8N6 and P8N9, respectively, with 48.72 ±3.07, 52.74 ± 2.35, and 39.61 ± 6.30% . Differently, in 10% PDLLA-containing scaffolds,the inclusion of additives first provoked a drop in porosity comparedto P10 (53.50 ± 3.89%). Then their crescent quantity induceda rise with P10N3 (34.29 ± 4.89%) lower than P10N6 (41.42 ±4.69%) and P10N9 (52.26 ± 5.01%). Statistical analysis showeda relevant difference only among P10, P10N3, and P10N9 . The effect of nanocrystals for 12%w/V PDLLA scaffolds was a gradual increment in porosity compared toP12 (37.11 ± 1.00%), followed by a decline: P12N3, P12N6, andP12N9 exhibited, respectively, values of 50.18 ± 3.26, 53.99± 1.50, and 45.19 ± 3.23%, with a statistical relevanceamong P12, P12N3, and P12N6 .Morphologicalanalysis indicated an inverse relationship betweenmean diameters and scaffold porosity -lactide onesas a term of comparison, whereas 8% w/V PDLLA and relative nanocompositescaffolds were discarded due to a large number of beads in the fibers.Membranes with the best morphological features were obtainedwith12% w/V PDLLA mixtures and, for this reason selected, for furtherstudies, using 10% w/V of poly- compared to the pure P12 (500 ± 50%), the increasein the doping by CNCs corresponds to an increase in the electrolyteuptake.Overall, all uptakes measured by swelling the membraneswith theelectrolyte are due to the strong affinity of CNC moieties with ions.64 SuperP carbon electrodes castedon copper or aluminum were used as working electrodes, while Li foilsas counter electrodes and LP30 as the electrolyte. Celgard separatorswere used as a comparison for all electrochemical tests. In order to measure the electrochemicalstability window of the scaffolds, LSV and CV tests have been carriedout.Focusing on the anodic scan 6a, the PFigures S6 and S7. The expected irreversible feature around 0.5 V vs Li+ is observed in both series of membranes in the first cathodic polarizationand, it disappears in the subsequent cycles. This peak originatesfrom the inevitable reduction of the electrolyte solvent species toform the stable solid electrolyte interface (SEI) over the carbonaceouscomponents of the working electrode. The stability of the formed SEIis demonstrated by the disappearance of the 0.5 V vs Li peak: in fact,in the second and the third cycles, only the peaks of the lithiumintercalation in the SuperP carbon are visible.65 Apparently, all scaffolds show very similar electrochemicalactivity in the CV tests without remarkable effects originated bythe addition of CNCs.Turning to the cathodic branch, Starting from LSV and CV data, the electrochemicalstability windows(ESWs) of the different membranes can be estimated see 2.4, due to the smaller onsetanodic limit.Overall, our estimates of the ESW suggests a straightforwardapplicabilityof the P10 series in LIBs with standard formulation  whereas the P12 series requires the use of a low-voltagepositive electrode, like LiFePO67 In this respect, the impedance responses of symmetric Li/Li cellshave been measured for 24 h at a temperature of 30 °C. The spectrafor all samples and Celgard separator are shown in Figure S8. Starting for the EIS spectra, it is possible toevaluate the cell ohmic resistance (or electrolyte resistance) thatis mainly related to the ionic transport across the separator,68 as well as the charge transfer resistance (orinterface resistance), that is due to the deposition/stripping oflithium to/from the metallic surfaces.Besides ESWs, the chemical stability of separatorstoward lithiumis another relevant performance parameter for LIB applications.The ohmic resistanceas well as the charge transfer resistanceis constant in an ideal Li/Li symmetric cell, and therefore, theirmonitoring is useful to highlight the occurrence of spontaneous chemicalparasitic reactions between lithium, electrolyte, and separator thatmay impact the motion of ions in the electrolyte and the electrokineticsof the plating/stripping. The obtained resistances as a function ofstored hours are reported in The additive-free P10 membrane shows a remarkablyconstant electrolyteresistance upon time, very close to the commercial Celgard. The additionof the CNC additive leads in variations of the electrolyte resistanceevolution upon time: it is apparently quite similar for the P10N3scaffold, whereas it shows smaller relative values compared for theP10N6 and the P10N9 ones. In these last two cases, this trend maysuggest a decrease in the tortuosity with the addition of CNCs.Turning to the P12 series, the behavior of samples is less homogeneous.Only P12N6 shows an evolution of the electrolyte resistance betterthan the Celgard separator, whereas P12 and P12N3 have resistancevalues close to the commercial separator. P12N9 suffers a sudden hugeincrease in the electrolyte resistance even only few hours of directcontact with lithium. Furthermore, stable electrolyte resistance valuesupon time are observed only for the P12N6 membrane.On the otherhand, Overall, theelectrochemical stability tests suggest that onlythe P10, P10N6, and P12N6 membranes are suitable as separators forlithium metal-based cell applications whereas in all other cases,unstable interfaces form leading to large and scattered values ofthe lithium/(electrolyte & separator) interface resistances.Among all the tested separators, the P10N6 formulation has beenselected for further preliminary electrochemical tests in a full Li-ioncell configuration.–2) and a commercial electrolyte (1 M LiPF6 EC:DMC 1:1 v/v)have been used in order to check the applicability of the P10N6 separator.Performance test results are shown in Commercial composite positive and negativeelectrodes, namely NCAand graphite (nominal capacity 3.5 and 3.8 mAh cmIn the present work, a set of electrospunmembranes to be employedas separators in aprotic LIBs has been developed with different amountsof PDLLA and CNC fillers. The morphology of the nanofibrous membranesimproved proportionally to the quantity of polymers.Among theevaluated separators, the P10 series exhibited the mostencouraging results, with neat P10 and P10N6. They displayed homogeneouselectrolyte resistance and electrochemical stability, and their electrolyteuptake was empowered gradually with the CNCs embedding empowered electrolyteuptake, gradually with the nanofiller content. For a practical evaluationof a hybrid scaffold, once assembled in a full Li-ion cell configurationusing commercial electrodes and electrolytes, the P10N6 separatorsuccessfully delivered performance in terms of reversibility and rateresponse. These findings all suggest the applicability of the PDLLA-basedseparators for lithium-based cells.


Despite advances in diagnosing and treating chronic heart failure (HF), the underlying mechanisms in different HF phenotypes remain unclear. Mitochondrial energy metabolism is crucial in HF etiology. Our study aimed to explore the value of metabolic-associated biomarker peroxisome proliferator-activated receptor-γ coactivator-1α (PGC1α) in identifying different HF phenotypes.N = 46), heart failure with reduced ejection fraction , heart failure with mildly reduced ejection fraction , and heart failure with preserved ejection fraction  groups. Each participant’s baseline data were recorded, blood samples were taken, and echocardiography was conducted. The level of PGC1α expression was determined using an enzyme-linked immunosorbent assay (ELISA) kit. The receiver operative characteristics (ROC) curve was further established in the four groups to assess the diagnostic value for overall HF and each HF phenotype with the calculation of the area under the curve (AUC) and 95% confidence interval (CI).A total of 172 participants were enrolled in the Affiliated Hospital of Xuzhou Medical University and were subsequently divided into four groups based on the European Society of Cardiology HF management guideline: the non-HF control  compared to non-HF participants (233.3 ± 32.69 nmol/L). Considering different HF phenotypes, PGC1α expression was considerably higher in the HFmrEF group (401.6 ± 45.1 nmol/L)than in the other two phenotypes .Furthermore, the AUCs of PGC1α in overall HF and each HF phenotype were all over 0.8, showing the ideal diagnostic value. Additionally, we provided the cut-off criteria for clinical use, which needs further validation. There was no significant correlation between PGC1α and N-terminal (NT)-prohormone B-type natriuretic peptide (BNP)/blood glucose, suggesting that PGC1α might exert a unique function in HF yet in a different pattern.We discovered that PGC1α could be used as a potential biomarker for differentiating HF patients from those without HF and for distinguishing HFmrEF from HFrEF and HFpEF. As one of the most stumbling blocks in the cardiovascular field, chronic heart failure (HF) remains a clinical and public health problem with high morbidity and mortality . In receThe current guidelines propose that HF diagnostic process should be constructed based on these aspects: left ventricular ejection fraction (LVEF) as determined by echocardiography, blood N-terminal pro-B-type natriuretic peptide (NT-proBNP) or BNP level, and clinical signs or symptoms of the patients. HF phenotypes can be divided into three main categories: heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), and heart failure with mildly reduced ejection fraction (HFmrEF) . HoweverMetabolism is crucial in the pathophysiology of cardiovascular diseases, particularly HF . The eneAdenosine 5′-monophosphate-activated protein kinase (AMPK)/Sirtuin 1 (Sirt1)/peroxisome proliferator-activated receptor-γ coactivator-1α (PGC1α) is a critical pathway in cardiac metabolism. Li et al.  have fouThe clinical study was constructed according to the Declaration of Helsinki, and the study protocol was approved by the Ethics Committee of the Affiliated Hospital of Xuzhou Medical University .A total of 172 participants were recruited for the study from May 2021 to December 2021 and divided into four groups  based on the inclusion and exclusion criteria  displayeWe recorded the participants’ demographic data  and drug use , measured several items , collected data on cardiovascular disease events , assessed cardiac function using echocardiography and NYHA classification, and performed blood biochemistry tests.Transthoracic echocardiography was performed on all participants using a commercial echocardiographic device . Measurements of left ventricular diastolic function were performed according to the guidelines. Trans-mitral early (E) and mitral annular early (e′) diastolic velocity indices were obtained, and E/e′ ratio was calculated to reflect diastolic function.Whole blood samples were collected, and routine laboratory tests  were performed in the clinical lab of the Affiliated Hospital of Xuzhou Medical University according to the manufacturer’s instruction. A dedicated kit-based NT-proBNP assay  was used to measure plasma NT-proBNP levels.PGC1α expression was quantified using an enzyme-linked immunosorbent assay (ELISA) kit . In brief, we added serum, standards, and HRP-labeled detection antibodies to pre-coated PGC1α antibody-coated micropores. For color rendering, 3,3′,5,5′-tetramethylbenzidine (TMB) was used after incubation and thorough washing. TMB is converted to blue by peroxidase and to yellow by acid finally. A positive correlation was observed between color depth and PGC1α in the samples. We measured the absorbance [optic density (OD) value] at 450 nm with a microplate reader and finally calculated the sample concentration.To evaluate the diagnostic value of PGC1α in identifying HF and each HF phenotype, the Receiver operative characteristics (ROC) curve was established using “pROC” R package (R version 3.6.3). To assess the sensitivity and specificity of the ROC curve, the area under curve (AUC) and 95% confidence interval (CI) were calculated via the “ggplot2” R package.post-hoc test. Data with a normal distribution were represented by mean ± standard deviation (SD), whereas median [interquartile range (IQR)] was used to represent data of a non-normal distribution. The χ2 test was used to compare categorical variables that were presented as numbers and percentages. P < 0.05 was considered statistically significant.Data were analyzed using SPSS software Version 20.0 ; GraphPad Prism Version 8.3.0 . Regarding continuous variables, initial analyses of the normality were conducted using the Kolmogorov-Smirnov (KS) test in conjunction with Q-Q plots . DiffereFurthermore, we assessed all participants by echocardiography. We mainly focused on LVEF and E/e′ ratio to assess cardiac systolic and diastolic functions. The results showed that LVEF (mean ± SD) in the non-HF, HFrEF, HFmrEF, and HFpEF groups were 62.13 ± 4.064, 33.33 ± 4.979, 43.73 ± 2.208, and 58.24 ± 5.057, respectively, which was consistent with the guidelines. Also, the E/e′ ratio for the four groups was 11.43 ± 5.084, 20.84 ± 12.92, 16.25 ± 5.437, and 19.6 ± 8.486, respectively, indicating that the diastolic cardiac function was impaired in HF.N = 46 for the non-HF group, N = 54 for the HFrEF group, N = 22 for the HFmrEF group, and N = 50 for the HFpEF group). There was no difference in terms of gender or height. Intriguingly, several items showed significant differences among the four groups. HF patients had a higher coronary heart disease rate and used more angiotensin-converting enzyme inhibitor (ACEI)/angiotensin receptor blocker (ARB) and diuretic drugs. Regarding blood biochemistry tests, HF patients had lower hemoglobin, higher creatinine, and higher C-reactive protein levels, indicating that HF patients are prone to inflammation when compared with non-HF controls.As shown in The ROC curve analysis showed that the AUC of PGC1α for the diagnosis of HF compared to non-HF was 0.843 (CI: 0.785–0.900), and the cut-off criterium was 293.918 nmol/L, suggesting that 293.918 could be used as the criterion for distinguishing HF from non-HF. Additionally, the AUC for diagnosing HFrEF, HFmrEF, and HFpEF compared to non-HF was 0.804, 1.000, and 0.815, respectively. The above-mentioned results revealed the ideal diagnostic value in identifying HF and its phenotypes from non-HF controls . FurtherCompared to the most widely accepted categories of HF (HFrEF and HFpEF), HFmrEF represents a borderline group that has received little attention, lacking effective diagnostic methods and treatment . HoweverPGC1α has been initially identified as a cofactor for nuclear hormone receptor peroxisome proliferator-activated receptor gamma (PPARγ) in adipocytes, which is required for the adaptive thermogenic response to a lower temperature . It reguIn our study, we revealed that PGC1α had the highest expression level in the HFmrEF group, which could be considered the transitory stage from HFpEF to HFrEF. PGC1α participates in mitochondrial biosynthesis ; hence, The value of PGC1α in diagnosing HF and identifying HF phenotypes is important, as shown on the ROC curve. The AUCs of the four groups were all greater than 0.8. Furthermore, AUC and Youden index were equal to 1in HFmrEF, showing the highest sensitivity and specificity in diagnosing HFmrEF. Meanwhile, we established a detailed threshold of PGC1α in diagnosing different HF phenotypes, which could be helpful for clinical practice. Additionally, we conducted the correlation analysis using Pearson test to investigate the relationship between PGC1α and blood glucose/NT-proBNP in HFmrEF patients. The results showed that there was no statistically significant correlation between PGC1α and blood glucose/NT-proBNP in HFmrEF group . TherefoThe newest AHA/ACC/HFSA Guideline for the Management of Heart Failure  publisheWe discovered that PGC1α expression was significantly upregulated in HF patients compared with non-HF participants. Besides, HFmrEF patients had a higher PGC1α expression level than HFrEF and HFpEF patients, which could be employed as a potential biomarker for differentiating HF patients from those without HF, as well as for distinguishing HFmrEF from HFrEF and HFpEF.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The study protocol was approved by the Ethics Committee of the Affiliated Hospital of Xuzhou Medical University  . The patients/participants provided their written informed consent to participate in this study.SZ and YH contributed to hypothesis development and manuscript preparation. SZ and YZ contributed to the study design, drafted and revised the manuscript. SZ was responsible for collecting clinical samples. YM and ZL undertook data analyses. All authors contributed to the article and approved the submitted version.


While oxygen-glucose deprivation (OGD) has been widely utilized in many cell lines to mimic certain biological changes, it has yet to be validated in mesenchymal stem cells. We performed RNA sequencing on adipose-derived stem cells (ADSCs) under hypoxic and glucose-free conditions after 4 h and 8 h. A total of 335 common differentially expressed genes (DEGs) were identified in the two OGD groups compared with the normal control group, consisting of 292 upregulated and 43 downregulated genes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that DEGs are mainly involved in metabolic processes, programmed cell death, and DNA-binding transcription activator activity. Protein‒protein interaction and hub gene analysis revealed various potential hub genes, in which response to oxygen levels, the IL-17-related biological function and the hypoxia-inducible factor 1 signaling pathway have been of vital importance. In summary, changes in transcription factor activity may play pivotal roles in oxygen-glucose deprivation. Through RNA sequencing, we have a deeper understanding of the changes in ADSCs after OGD treatment, providing more precise insight into predicting and regulating the stemness of ADSCs. Hypoxia is a fundamental component of the microenvironment and is associated with cell proliferation, cell differentiation and cell death. Appropriate hypoxic preconditioning of ADSCs can boost cell proliferation , promotein vivo  on ADSCs after different durations of OGD pretreatment to explore the functional impact of oxygen and glucose levels on ADSCs at the cellular level to obtain a more comprehensive understanding of possible changes in cellular biological activities during the early stage of transplantation.2, 5% CO2, and 94% N2.We harvested and identified ADSCs and built an OGD model as previously described . Briefly2, 5% CO2, and 74% N2; (b) OGD-4 h: cells with OGD treatment for 4 h; (c) OGD-8 h: cells with OGD treatment for 8 h.The cells were divided into three groups: (a) NC: normal control group cells were maintained in normal culture medium with 21% O2 or in MSCM in a normal incubator as a control. After 4 h and 8 h, 10 μl of CCK-8 reagent was added to each well and incubated at 37 °C for 2 h. The absorbance of each sample, which was proportional to the activity level of cell proliferation, was measured at a wavelength of 450 nm using a microplate reader. Each group was prepared in triplicate. The experiment was repeated three times.Cell Counting Kit-8  was used to detect cell proliferation according to the manufacturer’s protocols. Briefly, 10,000 cultured cells were seeded in 96-well plates overnight and then cultured in glucose-free DMEM under 1% OAnnexin V/7-AAD double staining was performed to analyze the state of the cells under different OGD treatment times. Briefly, three groups of cells were harvested, centrifuged, and resuspended in PBS at a concentration of 1 × 10/ml. The cell suspension was incubated with annexin V conjugated with FITC  and 7-AAD  for 15 min in the dark and loaded on a flow cytometer.Total RNA was isolated from the control, OGD-4 h, and OGD-8 h groups using the TRIzol method. The total amount and integrity of RNA were assessed using the RNANano 6000 Assay Kit of the Bioanalyzer 2100 system . RNA purity was evaluated by calculating the A260/A280 ratio, which should be between 1.8 and 2.0.Total RNA was used as input material for the RNA sample preparations. Briefly, mRNA was purified from total RNA by using poly-T oligo-attached magnetic beads. First strand cDNA was synthesized using random hexamer primers and M-MuLV Reverse Transcriptase, while second strand cDNA synthesis was subsequently performed using DNA Polymerase I and dNTPs. After adenylation of the 3’ ends of DNA fragments, adaptors with hairpin loop structures were ligated to prepare for hybridization. To preferentially select cDNA fragments 370–420 bp in length, the library fragments were purified with an AMPure XP system . Then, PCR amplification was performed, the PCR product was purified by AMPure XP beads, and the library was finally obtained.After the library was constructed and qualified by Qubit 2.0 Fluorometer, the different libraries were pooled according to the effective concentration and the target amount of data off the machine and then sequenced by the Illumina NovaSeq 6000 . The image data measured by the high-throughput sequencer are converted into sequence data (reads) by CASAVA base recognition. Clean data were obtained by removing reads containing adapters, reads containing N bases, and low-quality reads from the raw data. The index of the reference genome was built using HISAT2 (v2.0.5), and paired-end clean reads were aligned to the reference genome using HISAT2. FeatureCounts (v1.5.0-p3) was used to count the read numbers mapped to each gene.via principal component analysis (PCA)  . The R pnge| > 1 . The orilanguage .p value < 0.05 were selected for further analysis. In addition, the ggplot2 and GOplot packages were used for cluster analysis. Networks were also generated in the Cytoscape software environment with the EnrichmentMap plugin using an uncorrected p value threshold of 0.005, an FDR cutoff of 0.1, and an overlap coefficient threshold of 0.1.The biological functions of the shared DEGs were explored with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses . GO is tProtein–protein interaction (PPI) analysis was performed using an online database named Search Tool for the Retrieval of Interacting Genes (STRING). DEGs with a minimum required confidence score ≥0.7 were chosen to build a full network model, which was then visualized in Cytoscape software. In the network, each node corresponds to a protein, and each edge represents an interaction. MCODE, a molecular composite detection plugin in Cytoscape, was used to filter out hub genes from numerous candidates. The parameters were set as follows: Degree Cutoff = 2, Node Score Cutoff = 0.2, K-Core = 2, Max. Depth = 100. The most common and largest module was defined as having an MCODE score >4.via the clusterProfiler package.Hub genes were screened by the combination of 12 prediction algorithms in the Cytoscape plugin cytoHubba . For eacp < 0.05.All quantitative results are presented as the means ± standard deviations (SD). Statistical comparisons were performed using one-way ANOVA in three independent experiments. GraphPad Prism version 9.0 software  was used for data analysis. Statistical significance was set at After being treated with OGD for 4 h or 8 h, some ADSCs were separated from the bottom of a culture dish, and the number of floating cells was measured . OGD trein vivo, we cultured ADSCs under hypoxic and glucose-free conditions and performed high-throughput sequencing at three different time intervals  to further identify the underlying mechanism in the pathophysiological process. After filtering the original data, checking the sequencing error rate, and verifying the distribution of GC content, clean reads for follow-up analysis were obtained. The 4 h samples OGD4h_1, OGD4h_2 and OGD4 h _3 generated 42.36, 40.77 and 45.51 million clean reads, while the 8 h samples OGD8h_1, OGD8h_2 and OGD8 h _3 generated 52.28, 45.78 and 46.14 million clean reads, respectively. Normal control samples NC_1, NC_2, and NC_3 generated 42.83, 42.97, and 39.58 million clean reads, respectively , CDKN1B, CRKL, CYCS (CytC), EGFR, FOS, FOXO1, HIF1A, IL6, JUN, MYC (C-Myc), NGF, NOTCH1, NOTCH3, PDGFRB, PXN, RORΑ, TGFB1, TLR4, VEGFA and WDR12, were identified as hub genes in in vivo, we speculate that hypoxia and a low glucose level were associated with stemness maintenance and contributed to the decreased cell number and increased apoptosis. Therefore, we searched for genes with potential roles in ADSCs under oxygen-glucose deprivation. Three time points  in the oxygen-glucose deprivation phase were chosen, and the high-throughput sequencing data identified hub genes guiding cell proliferation and death.Most of the studies exploring oxidative stress and glycometabolism investigated the proliferation and death of ADSCs exposed to hypoxia without glucose level modification . Given tp values. These findings suggest the importance of transcription factors in the OGD model.At two time points of oxygen-glucose deprivation, gene expression analyses revealed a higher number of differentially expressed genes at 8 h. A total of 335 DEGs overlapped between the 4 h group and the 8 h group , includiIn the identification of key modules and hub genes, JUN, IL6, CXCL1, VEGFA, TLR4, PTGS2 (COX2), RORΑ, KDM3A, KDM4B, KDM6B, KDM7A, EGFR, NOTCH1, JUN, HIF1A, FOXO1, RORΑ, CDKN1A (p21), and CDKN1B (p27) were characterized with the highest score , and theIn general, through bioinformatic research methods, we identified genes differentially expressed in the cytopathological process of hypoxia and hypoglycemia compared with the normal group and explored their potential function and relevant pathways. Changes in transcription factor activity and the HIF-1 pathway may play pivotal roles, and further experiments are required to further clarify the effect of these hub genes.


For all ear fasciation-related traits, fasciation level positively correlated with KRN (0.30 ≤ r ≤ 0.68). Prolificacy and tillering were not correlated and their h2 ranged from 0.41 to 0.78. QTL mapping identified four QTLs for ear fasciation, on chromosomes 1 (two QTLs), 5 and 7, the latter two overlapping with QTLs for number of kernel rows. Notably, at these QTLs, the Lo1016 alleles increased both ear fasciation and KRN across populations, thus showing potential breeding applicability. Four and five non-overlapping QTLs were mapped for ear prolificacy and tillering, respectively. Two ear fasciation QTLs, qFas1.2 and qFas7, overlapped with fasciation QTLs mapped in other studies and spanned compact plant2 and ramosa1 candidate genes. Our study identified novel ear fasciation loci and alleles positively affecting grain yield components, and ear prolificacy and tillering loci which are unexpectedly still segregating in elite maize materials, contributing useful information for genomics-assisted breeding programs.Maize ear fasciation originates from excessive or abnormal proliferation of the ear meristem and usually manifests as flattened multiple-tipped ear and/or disordered kernel arrangement. Ear prolificacy expresses as multiple ears per plant or per node. Both ear fasciation and prolificacy can affect grain yield. The genetic control of the two traits was studied using two recombinant inbred line populations (B73 × Lo1016 and Lo964 × Lo1016) with Lo1016 and Lo964 as donors of ear fasciation and prolificacy, respectively. Ear fasciation-related traits, number of kernel rows (KRN), ear prolificacy and number of tillers were phenotyped in multi-year field experiments. Ear fasciation traits and KRN showed relatively high heritability ( OntogenArabidopsis, SAM homeostasis was shown to be controlled by the CLAVATA3 (CLV3) - WUSCHEL (WUS) feedback signaling pathway , causing the decline of stem cell proliferation, and a corresponding decrease of CLV3 production (CLV3/EMBRYO-SURROUNDING REGION7 (ZmCLE7) is a CLV3 ortholog; THICK TASSEL DWARF1 (TD1) and FASCIATED EAR2 (FEA2) encode receptor-like proteins related with CLV1 and CLV2, and regulates the size of the ear IM and SAM growth; ZmWUS1 seems to be a direct WUS ortholog and is expressed in the late vegetative SAM (ZmFEA4 encodes a basic leucine zipper domain (bZIP) transcription factor that is expressed in the shoot meristem peripheral zone and is likely involved in cell transiting from SAM to organ primordium (unbranched2 (UB2) and unbranched3 (UB3) are expressed in the initiating leaf primordia and the base of the SAM and control lateral organs initiation (ramosa1 (ra1), ra2 and ra3 , encoding a homeodomain leucine zipper transcription factor , encoding a bHLH transcription factor orthologous to rice LAX PANICLE1 , a transcriptional regulator with an AT-hook DNA binding motif  RIL populations, B73 × Lo1016 and Lo964 × Lo1016 analyzed as a joint population. Line Lo1016 was the genetic source of mild ear fasciation while Lo964 was the source of ear prolificacy.22.11s. Both Lo964 and Lo1016 are typical dent lines originally bred at the Bergamo breeding station , shown to be genetically related based on molecular marker analysis and classified as BSSS = Iowa Stiff Stalk Synthetic heterotic group  and Lo964 × Lo1016 (L×L) populations eventually included 97 and 68 RILs, respectively. We will refer to the Joint Population (JP) as the assembly of the whole set of 165 (97 + 68) RILs.Two recombinant inbred line (RIL) populations were developed as follows. The line Lo1016 was crossed with B73 and with Lo964 to create two Fic group . B73 is ic group . Single 2.2Field trials were carried out in Cadriano, near Bologna, Italy  in 2017 and in Monselice, near Padova, Italy  in 2018 and 2019. Field experiments were organized as a randomized complete block design with two replicates (one rep = one plot with 10 plants per RIL). Plots for each of the three parental lines were included in the experiment. Plot length was 2.5 m, distance between rows was 0.8 m and between plants was 0.25 m. Plots were overplanted by hand and thinned at the V7 growth stage to one plant per hill equivalent to 11 plants per plot, and with an overall investment of 5.5 plants per square meter. The field was managed following standard agronomic practices of the area.Phenotyping for ear fasciation was carried out by collecting four traits, namely ‘ear ovality’ ; ‘kernel row disorder’ ; ‘ear diameters ratio’ ; ‘ear fasciation index’ . Visual scores per ear were given by three persons independently, and mean values were utilized as entries for subsequent analysis. Number of kernel rows (KRN) was collected by counting the number of kernel rows at mid-ear position, on the same ears subjected to phenotyping for fasciation. Plant architecture traits collected were number of tillers per plants (TIL) and proportion of plants per plot showing prolificacy (PROL), i.e., proportion of plants showing >1 ear at the top ear node. For all traits, three central contiguous plants per plot were considered.r’ instead of the more appropriated ‘rho’ for clarity in text. Trait distributions were normal or reached normality after square root transformation  in the R package “tidyverse” . BLUES vormation , except Analysis of variance (ANOVA) was run for six out of seven traits and for two  out of three years. Specifically, ANOVA was not run for ear fasciation index since this phenotypic dataset was only available as mean value of two repetitions. Similarly, phenotypic data for year 2017 were not included in ANOVA because mean values (as mean of two repetitions) were available only. ANOVA output tables for six traits are provided in 2.3P ≤ 0.05 with 1,000 permutation). QTL additive effects were always computed by the formula 2a = (mean homozygous B73 – mean homozygous Lo1016) or 2a = (mean homozygous Lo964 – mean homozygous Lo1016), for the two RIL populations, respectively.B×L and L×L were genotyped using a high density 15-K SNP array  using a qProl1 and qProl4, for which we provided footnotes in Additionally, QTL mapping for ear diameters rate, ear prolificacy and tillering  was also carried out by Kruskal-Wallis (KW) test, known to be robust to deviation from distribution normality, as implemented in MapQTL6 . For the2.4www.maizegdb.org/nomenclature.QTL confidence intervals from this study were projected on B73 reference genome B73v5.  and incl33.1vs 16.33 or 14.54, for B73 or Lo964, respectively, P < 0.01). Lo964 showed the highest ear prolificacy among the three parental lines . In addition, Lo1016 was the only parental line developing tillers (ca. 1.5 tillers per plant). Values for additional plant architecture traits recorded in this study are reported in P < 0.01) while ear ovality and number of tillers were only mildly affected (0.01 < P < 0.05) (P < 0.01) except number of tillers. No ‘Population-Year’ interactions were observed for any traits with the exception of prolificacy. For any trait, RIL (ie. lines) was a major source of variation, in line with the presence of segregating QTLs. Broad sense heritability (h2) for ear-fasciation traits ranged from 0.13 for ear diameters rate in L×L to 0.95 for ear fasciation index in B×L. Ear prolificacy and TIL h2 were relatively high  with KRN and ear diameters rate the lowest  resulted significantly correlated, with 3.2The three linkage maps, namely B×L, L×L and JP included 1,186, 984 and 1,303 markers, and covered 1,819.52 cM, 2,504.5 cM and 1,661.0 cM, respectively  within two narrow supporting intervals of < 2 Mb, and both segregated within B×L only. qFas5 was mapped on bin 5.07 for fasciation index and ear ovality and appeared to segregate mainly in L×L. qFas7 was mapped on bin 7.02 and shown to affect ear fasciation index, ear ovality and kernel disorder in both B×L and L×L. Notably, for all qFas QTLs, the fasciation-increasing allele was provided by Lo1016, the parental line showing ear fasciation  will be considered as the same QTL whenever their supporting intervals overlap, considering QTL results from B×L, L×L and JP  of phenotypic variation with a genetic effect 2a = 0.69 or 0.87 kernel rows, in JP or L×L, respectively . The QTL qKRN5 on bin 5.07 controlled 10% of phenotypic variance and showed an effect of 2a = − 0.57 kernel rows . Two KRN QTLs, qKRN7.1 and qKRN7.2, mapped on bin 7.02 and 7.03, respectively, and had similar genetic effect . The QTL qKRN8 mapped on bin 8.02, with a genetic effect of 2a = −0.79 in L×L . Notably, all KRN QTLs segregated in L×L while none in B×L. Overall, qKRN2 was the only KRN QTL with the ‘+’ allele contributed by Lo964, while Lo1016 contributed the ‘+’ allele at the other four KRN QTLs  also increased kernel row number, supporting the hypothesis of a functional association due to the presence of causative gene(s) affecting both ear fasciation and number of kernel rows, and in line with the observed positive correlation between the two traits  and qProl9 (16.7%). At qProl1, qProl2 and qProl4, the high ear prolificacy parent Lo964 contributed the ‘+’ QTL allele. Both qProl1 and qProl2 showed narrow physical supporting intervals . Tillering variation was also shown to be under polygenic control with four QTLs. The two strongest QTLs in terms of genetic effect, qTil1 and qTil2, mapped on bin 1.05 and 2.01, controlled 12-13% of phenotypic variance in JP with a genetic effect of 2a = 0.6 tillers per plant. Although the ‘+’ allele was contributed by Lo1016 for qTil1, qTil4 and qTil9, in both B×L and L×L B73 contributed a positive tillering allele at qTil2. No overlap was found between prolificacy and tillering QTLs.Four QTLs were mapped for ear prolificacy (3.3.4www.maizegdb.org) along with gene expression information in meristem and ear primordium. Alongside, a list of 42 genes involved in development and/or proliferation of ear meristem was collected by screening the scientific literature , compact plant2 . Additionally, at qKRN8 , barren inflorescence1  and arftf3  were identified within the supporting interval of qTil2, on chr. 2.In order to search for candidate genes of ear fasciation QTLs identified in this study, we extracted all gene models included in the QTL supporting intervals present in B73 v4  or based on the de novo whole genome shotgun sequences obtained in this work (Lo964 and Lo1016), and compared in order to identify functional variants. Specifically, variants were searched for ra1 (candidate at qKRN7.1/qFas7), ct2 and bgh1 (candidates at qFas1.2) and crr1  and the presence of QTLs affecting only one out of four ear-fasciation-related traits  confirmed that ear fasciation is a genetically and physiologically complex polygenic trait and that at least some genes can possibly affect kernel disorder without affecting cob ovality/flatness, or vice versa. The presence of QTLs specific for single components of ear fasciation was already reported , as previously hypothesized or shown. Notably, at both qFas5-qKRN5 and qFas7-qKRN7.1 the fasciation/KRN-increasing allele was provided by Lo1016, and the effect of the Lo1016 alleles at the two QTLs was detected in both B×L and L×L. In other words, at both qFas5-qKRN5 and qFas7-qKRN7.1, Lo1016 carries alleles increasing both ear fasciation index and kernel row number, and their positive effect on ear fasciation were detected across genetic backgrounds. However, the KRN effect was detected in the L×L background only, likely because the KRN mean value in the L×L genetic background was lower than that in B×L  by qFas1.2 mapped in our study. qKRN5-4) between umc1971 and umc1071 on chr. 5 affecting kernel row number, and mapping nearby our qFAS5. Finally, the chr. 7 region corresponding to our qFas7-qKRN7.1 appears as an ear fasciation/KRN QTL hot spot. QTLs mapped in this region included cob and ear flatness qCF7 and qEF7 by Mei et al. (2021), ear fasciation fa_c1 by qkrn7 by KRN7.1 by Chen et al. (2019), qKRN7 by Our QTL consensus map  known to be involved in ear prolificacy , a known gene that when mutated produces barred shoots with no ear  had the tillers-increasing allele provided by Lo1016, in line with the phenotype of parental lines (Lo1016 is the only parent showing tillering when grown in the field in standard conditions). However, qTil2 had the tiller-increasing allele contributed by B73 that develops hardly no tillers in our field conditions  perfectly overlapped with an ear prolificacy QTL formerly identified in a maize × teosinte cross. Quite unexpectedly, we did not find correlation or QTL overlaps between ear prolificacy and tillering, although the two traits share obvious developmental basis.Overall, our study provides clear entry points for the molecular dissection of important yield component traits, which should help both developing molecular markers for marker-assisted selection to be deployed in breeding programs and starting the procedures leading to cloning the genes underpinning the QTLs described and eventually their manipulation by engineering or editing.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.SS and RT conceived the project and acquired funding; KL, AT, SG, SR, CU and SS designed and curated field trials, collected phenotypic and genotypic data and analyzed the data; KL, RT and SS wrote the first manuscript draft. All authors contributed to the article and approved the submitted version.


Cupressus sempervirens, sometimes called Italian or Mediterranean cypress, is found in subtropical Asia, North America and eastern Mediterranean region. Pharmacological investigations of Cupressus sempervirens showed biological properties such as aromatherapeutic, antiseptic, astringent, balsamic or anti-inflammatory, astringent, antiperspirant, diuretic and antispasmodic. Chemical analysis of Cupressus sempervirens gives phytochemicals like monoterpenes, diterpenes, flavonoid glycosides and bioflavonoids. The current review highlights interactions, conventional uses and biological actions of Cupressus sempervirens plant and plant products.The roles of plants and its products in all forms of life cannot be overemphasized. The medicinal products from plant are phytochemicals, drugs, food supplements, beauty products, etc. In ethnomedicine, leaves, fruits, stem, bark, root and fluids from plants are used in the cure, management and prevention of several diseases.  Cupressus sempervirens revealed that essential oils, alkaloids 0.7%, saponins 1.9%, flavonoids 0.22%, phenols 0.067%, tannins 0.31%, and other bioactive compounds were present in the plant. Medical examination in the past found that Cupressus sempervirens contained antioxidant, anticancer, antifungal, antibacterial, antiparasitic, antiviral, insecticidal, anticoagulant, estrogenic, healing and numerous properties  A. Camus, Cupressus sempervirens L. variety sphaerocarps (Parl.) Parl., Cupressus sempervirens L. variety umbilicata (Parl.) Parl., Cupressus sempervirens L. Forma stricta (Aiton) Rehder, Cupressus sempervirens L. subspecies indica (Parl.) Silba, Cupressus sempervirens L. variety Atlantic (Gaussen) Silba, Cupressus sempervirens L. variety dupreziana (Camus) Silba, Cupressus sempervirens L. variety globulifera Parl., Cupressus sempervirens L. variety horizontalis (Mill) Loudon, Cupressus sempervirens L. variety indica Parl., Cupressus sempervirens L. variety numidica Trab., Cupressus sempervirens L. variety pendula (Endl.) A. Camus, Cupressus sempervirens L. variety stricta Aiton  from J. phoenicea and C. sempervirens were obtained respectively  of Moroccan leaves  (w / w) and α-terpinyl acetate (14.7%) as its second most important constituents  is the second most important component; this is similar to Moroccan J. phoenicea . These plants are directly used or can be processed by hydrodistillation (HD) to obtain essential oils or by extraction to obtain what must be termed ‘volatile fractions’. These oils are used in pharmacy, cosmetics, perfumes and food industry among others. The compounds identified in essential oils are thirteen in number. Mainly, the components of both the fruits and the leaves were -3-carene, α-pinene, terpinolene and α-terpenyl acetate, although the identified volatile oil is from Cupressus sempervirens L cv cereiformis. Leaves and fruit% respectively were: α-pinene 39.0 and 30.0, sabinene 3.0 and 2.0, β-pinene 2.2 and 2.6, myrcene 3.9 and 4.1, Δ-3-carene 24.0 and 24.0, limonene 3.0 and 4.0, terpinolene 4.3 and 6.6, traces and 1.7 of bronyl acetate, α-terpenyl acetate 6.6 and 5.6, 1.2 and traces of β-caryophyllene, 1.3 and traces of α-humulene, germacrene D 1.7 and 4.0, while grouped compounds: ; (monerpenes containing oxygen 7.3 and 6.6); (sesquiterpenic hydrocarbons 1.7 and 10.5); (sesquiterpenes containing 4.0 and trace oxygen)  and -hydroxybenzoic acid methyl ester (15.5%). Other important aglycones were p-cymene-8-ol (5.3–6.4%) and perilla alcohol (3.6–8.2%), and carvacrol (2.5–6.3%) and 2-phenylethanol (2.7–6.9%). Aglycones with glycosidic bonds and simultaneously free compounds found in essential oil were found to be different     and was used in the treatment of hemorrhoids, venous circulation disorders and varicose veins. The essential oil was utilized as an antispasmodic for persistent coughs and antiseptic , Staphylococcus aureus (ATCC6538), Bacillus subtilis (ATCC6633), Escherichia coli (ATCC15224), Pseudomonas aeruginosa (ATCC6643) and Salmonella typhimurium (ATCC13048). These extracts’ concentration was utilized . All Cupressus sempervirens extract administered showed a dose-dependent inhibition of bacterial development against all microbes examined , Staphylococcus aureus (Gram-positive), Escherichia coli, Pseudomonas aeruginosa and parasitic species of Aspergillus niger and Candida albicans. Cupressus sempervirens was used to assess the bactericidal and antifungal action of Cupressus sempervirens chloroform and water extracts’ high intensity against Gram positive microbes (inhibition zone of 9–12 mm and 9–14 mm and chloroform and water extracts respectively) and reduced action against Gram negative microorganisms (zone of inhibition 1–6 mm and 1–5 mm water extract and chloroform extract). In any case, the water extract did not show much potency against growths; however, the chloroform extract displayed slight action against Candida albicans (3 mm) . Methanolic extract shows huge antimicrobial activities followed by the ethyl acetate and ethanol extracts. The methanolic extract demonstrated greatest inhibitory activity against B. subtilis, K. pneumonia and S. aureus. The ethanolic extract showed its action against P. aeruginosa, while the ethyl acetic acid extract of Cupressus sempervirens demonstrated a more prominent inhibitory action against E. coli and S. typhimurium  and Gram negative ; the inhibitory zones have diameter of 4 to 12 mm, with MBC and MIC values ranging somewhere in the range of 62.5 and 250 μg/ml. In spite of the fact that Cupressus sempervirens methanolic extract firmly restricted the progression of many bacteria growth . The outcome revealed that Cupressus sempervirens oil restricted the bacteria development, yeast and filamentous growths. The MCC and MIC values showed that the Cupressus sempervirens essential oil remained exceptionally powerful. Therefore, the MIC/MCC proportion affirmed antibacterial and anti-fungi essential oil action. In spite of the fact that the antimicrobial action of Cupressus sempervirens essential oils was evident against Gram-positive compared to Gram-negative bacteria : Staphylococcus aureus 7 and 8; Micrococcus luteus 10 and 13; Mycobacterium simegmatis 10 and 11; Bacillus megaterium 7 and 9; Enterococcus faecalis 7 and 9; Streptococcus faecalis 7 and 9; Saccharomyces cerevisiae 9 and 10; Bacillus brevis 7 and 8; Pseudomonas pyocyaneus 9 and 11; Aeoromonas hydrophila 7 and 10; Yersinia enterolitica 8 and 9; and Klyveromyces fragilis 15 and 17 mm  and seven parasites . Essential oil inhibition zones for 96 h of brooding against Bifidobacterium lactis were 24.05 mm, Escherichia coli was 16.11 mm, while Micrococcus luteus was 11.90 mm. Concerning essential oil fungicidal action inhibition zones were 5.7 mm against F. solani and 29 mm against P. digitatum following 96 h of incubation  against methicillin-resistant Staphylococcus aureus , and all Cupressus sempervirens essential oil tests demonstrated astounding antifungal control against all growths examined . Antifungal in vitro tests were tried on the oils, and the effect revealed a serious growth inhibition of 10 plant pathogenic fungi  contamination was performed on HeLa cell monolayers to assess the antiviral property of plant extract under a light magnifying lens utilizing the hematoxylin and eosin technique. Test outcomes were estimated against acyclovir, a positive control. As indicated by the outcomes, antiviral properties against the HSV-1 infection were seen in the three plants. The strongest extract was that of C. sempervirens. Of the different parts tried, the organic product extract was appeared to have the highest anti-HSV action (92). The in vitro assessment done indicated that a small amount of proanthocyanidin polymer  was extracted from Cupressus sempervirens L.  that are treated with Cupressus sempervirens powder cones, gathered in Oxford, Mississippi, were examined. The utilization of centrifugal slim layer chromatography caused the isolation of many 13-abietatrien-12-one), 6-deoxytaxodione , diterpenes, taxodione, sugiol and ferruginol. 6-Deoxytaxodione , 13-abietatrien-12-one) of which taxodione indicated a solid antileishmanial action with values of a fraction—greatest inhibitory focus (IC50) of 0.077 μg/ml and 0.025 μg/ml, separately, against promastigotes of Leishmania donovani, compared with those of the standard antileishmanial drugs, pentamidine (IC50 1.62 μg/ml) and amphotericin B (IC50 0.11 μg/ml)  > acetone extract (LC50 104.3 ppm) > crude oil ether separates (LC50 37.8 ppm). A huge decrease was seen in both the level of pupae and the growth of the adults although all the extracts demonstrated a delayed lethal effect in pupae and grown-ups after larval treatment. Furthermore, diverse degrees of morphogenic deviations from the norm were seen in the young and adult stages  against radicals was 65% unlike the chloroform extract (50 μg/ml) which is 6% , reducing power test was 2.85 (mg of ascorbic acid/g of extract); via metal chelation the antioxidant action of two varieties of extracts was tried utilizing 2,2-diphenyl-1-picrylhydrazyl (DPPH) and N, N-dimethyl-p-phenylenediamine (DMPD) radical scavenging action, chelating capacity of metals along with ferric-(FRAP) and lessening antioxidant intensity of phosphorus-molybdenum (PRAP). The antioxidant activities test was done at 2000 µg/ml. From the exploration discovering, there was variety in antioxidant activities because of the technique utilized. For example, Cupressus sempervirens ethyl acetate cone extract var. horizontalis indicated greatest DPPH radical scavenging activities (87.53 ± 0.17%), while just six of the extracts had the capacity to displace the radical of DMPD with a range of 6.06 ± 0.23 and 30.34 ± 0 69%. The resultant effects of the FRAP test show that Cupressus sempervirens cone acetone extract var. horizontalis demonstrated the absorbance value at its peak showing elevated antioxidant action, despite that the extracts commonly had low action in the PRAP examined. Methanolic extract of the leaf of Cupressus sempervirens var. horizontalis has the elevated antioxidant action. The cone and leaf methanol extract lack metal chelating limit in the metal chelation test. Conversely, Cupressus sempervirens ethyl acetate foliar extracts var. horizontalis (75.86 ± 0.33%) and Cupressus sempervirens var. pyramidalis (77.07 ± 3.22%) indicated the maximum activity in this test. Cupressus sempervirens essential oil was examined for its antioxidant property by estimating the scavenging effect of the radical on 2,2-diphenyl-1-picrylhydrazyl (DPPH) and utilizing 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonate) (ABTS) paper. The test result showed elevated antioxidant activities  matched with BHT , the peroxyl radical-interceded red platelet hemolysis (RBC) and linoleic acid peroxidation test and test were utilized. The peroxidation of linoleic acid for 4 h was assessed every incubation hour. The anti-glycation action of the oils at 600, 400, 200 µg/ml was observed utilizing glycation surveys for hemoglobin and insulin. Hemoglobin glycation was restrained both by the twigs  and by the natural products  at 600, 400, 200 μg/ml of oil respectively. The inhibition rates of insulin glycation include  that of twig oil and  that of oil of natural products at 600, 400, 200 μg/ml, respectively. The red platelet hemolysis was further inhibited by twig oil  and natural product oil  at 260, 220, 180 μg/ml, respectively. Lastly, there is decreased linoleic acid peroxidation by the oils which arrived at its greatest point after 4 h for the two twigs  and natural products  at 260, 220, 180 μg/ml of oil separately; 600 and 400 µg/ml were assessed utilizing insulin and hemoglobin glycation surveys. The glycation of hemoglobin was inhibited both by the twigs  and by the organic products and  at 600, 400 and 200 μg/ml of oil respectively; the inhibition rates of insulin glycation were  for twig oil and  for organic product oil 600, 400 and 200 μg/ml, respectively. The twig oil inhibited red platelets hemolysis by  including natural product oil  at 260, 220, 180 μg/ml, respectively. To conclude, linoleic acid peroxidation was lessened by the oil; after 4 h the greatest point was reached for the two twigs  and also for organic products  at 260, 220 and 180 μg/ml of oil individually.  or extract  or) show the safety index in normal rats and hepatoprotective action against toxicity of paracetamol   ethanolic extract of the fruits of the soil activity given in its part rich in diterpene, solvent in chloroform. Eight essential diterpenes were gotten indicating mild to strong action with the strongest diterpene (labda-8 (17), 12,14-trien-19-oic acid) showed an IC50 of 37.5 μM (with antiproliferative action against BPH stroma cells in humans). There was a significant inhibition of Stat-3 enactment (phosphorylation) in BPH stromal cells and inhibited androgen-delicate KLK3/PSA transamination in LNCaP cells TMPRSS2 qualities. The CS fraction in acid12, 14-trien-19-oico, labda-8 (17), inhibited rat model prostatic hyperplasia thereby causing TUNEL marking in stromal cells with lower articulations of bcl portion—2/bax, IGF-I, TGF-ß and PCNA  was monitored. Utilizing the trial rat model, paracetamol was utilized to induce liver toxicity in rats, Cupressus sempervirens hydroethanolic extract were used to find out the hepatoprotective and anticancer activities of while using silymarin as reference agent. A 4-week study from previous researches has used silymarin  or hydroethanolic extract  or) showing hepatoprotective activities compared to one toxic dose of paracetamol (4 g/kg bw) and a good profile in normal rats  in Wistar rats was done with hypolipidemic effect. The extract orally administered caused a huge reduction in total serum cholesterol, which remain significant for a period of 6 weeks. After the beginning of treatment (p < 0.001), the animals were presented with lower cholesterol levels at (p < 0.05); there was a substantial decrease in serum fatty oils, contrasting 0 weeks and 6–24 weeks. It was seen that between the animals treated with CSE and the control there were no significant difference in fatty substance levels and HDL cholesterol level throughout the experiment  (positive control), another group got CCl4 and were treated with Cupressus sempervirens extract thrice each week for 6 weeks and a group got CCl4 for about 6 weeks and were allowed to recover by themselves. After the experiment, all animals were sacrificed after which biochemical and histological parameters were assayed. There was a clear difference across all the groups. The rats treated with CCl4 and left for 6 weeks to recover on their own exhibited mild enhancements in the parameters studied. Administration of plant extract improved the altered biochemical parameters to a large extent. The histopathological evaluation of the liver and kidney of the group treated with Cupressus sempervirens was closely related to that of control groups (Koriem Cupressus sempervirens flavonoids protective action (quercetin and rutin) against the toxicological effect of lead acetate on the liver was studied. Thirty male albino rats were distributed into five groups (six rats for each group). The control is group I; Group II got lead acetate of 0.5 mg/g in their feed for 60 days. Group III got 8 mg/100 g body weight of Cupressus sempervirens (lyophilized from methanol extract of seeds) 14 days before the administration of lead acetate. Group IV were given 0.3 mg/100 g of body weight of the flavonoid quercetin every day for 14 days before the administration of lead acetate; Group V were given flavonoid daily 0, 1 mg/100 g of body weight for 14 days before lead acetate. Lead acetate was found to cause elevated serum and tissue ALT, ALP, AST and tissue MDA, bilirubin, tissue and plasma NO, regardless of significant elevated serum cholesterol, fatty substances, LDL and HDL. Though, lead caused a significant reduction in all out serum and tissue protein, albumin/globulin proportion albumin, globulin, SOD and GPx in blood and tissue compared with the benchmark group. Treatment with methanolic extract of Cupressus sempervirens, quercetin and rutin for 14 days before lead acetate did not allow increase in parameters. In like manner, treatment with methanolic extract of Cupressus sempervirens and its flavonoids can increase the defensive activity against the poisonous effect of lead acetate  and var. pyramidalis (CSP) cones and leaves was performed to counter acetylcholinesterase (AChE), tyrosinase (TYRO) and butyrylcholinesterase (BuChE). The extracts demonstrated mild to adequate cholinesterase inhibition at 200 µg/ml. The CSP cone dichloromethane extricate demonstrated the maximum inhibition (36.10 ± 1.45%) against AChE, while the greatest inhibition (40.01 ± 0.77%) against BuChE was done by the CSH leaf acetone extract  was 0.2837 ± 0.0115 mg/ml . Among the four studied, it was observed that the dose of 1.0 mg/kg bw of sugiol showed osteoprotective effects significantly, but at the same dose there was not uterine estrogenicity. Additionally, there was an improvement of biomechanical properties, as demonstrated by increase energy, power and rigidity in the femoral bones in the group treated with sugiol compared to the Ovx animals that were untreated . The endothelium-derived nitric oxide-associated maximal unwinding (Emax) and EC50 approved for acetylcholine were notably higher in the pretreated group compared with the control. From the information, it was proposed isochemically tested rats pretreated with cypress water extracts build the suitability  in rats from endothelial cellular function in isolated aortic rings assessed in vitro. Also, it had anticoagulant properties. Because of these effects, their effect on the endurance of arbitrary augmentations of ischemic pivotal folds was analyzed. The previously treated group was treated for 7 days with 30% cypress cone water extract before a fold rise, another 3 days a short time later. The ischemic objective was a 6 × 7 cm islet epigastric supply route flap dependent on the inferior epigastric pedicle at the right. At the time of the conclusion of the research, hemodynamic factors, mean arterial pressure including the pulse rate were evaluated. Flap endurance and perfusion rates were assessed using laser Doppler flowmetry and microangiography. The aortic isometric pressure separated from the control and pretreated groups was extracted to mirror the vascular feedback. Dose–response connections to acetylcholine were resolved and placed side by side with the control group. Hemodynamic factors did not show any difference. In the group pretreated, micro angiograms caused prolonged angiogenesis, slender thickness and expanded folds’ perfusion (blood perfusion parts) in the proximal and distal parts  and orally ). From the outcome, it was discovered that 90.7% of the total amentoflavone circulated as metabolite that are conjugated after oral administration. The infusion ip and iv in the plasma of rats 70.2% ± 5.18% and 73.2% ± 6.29% became metabolites from amentoflavone administered altogether. Moreover, the compound bioavailability declined in the ip infusion (77.4% ± 28.0%) than oral intake 0.04% ± 0.01% , intravenous  and orally . Therefore, it was discovered that 90.7% of the total amentoflavone turned into metabolites after oral administration. The iv and ip infusion was seen in rats’ plasma in the placebo; 73.2% _6.29% and 70.2% _ 5.18% of the all amentoflavone were available as formed metabolites  crude extraction from roots as the major component (85.5%) was given to human subjects intravenously and through multiple infusion; the half-lives were − 1.2–1.3 h for paeoniflorin. Following the exposure to major organs  while its essential urging dynamic metabolite was accounted for peric acid , but urea increased (p £ 0.05) compared with the normal control group , but the control estimation of ALT (17.12 IU L-1) and AST (25.90 IU L-1), a huge increase in the degrees of 24.05 IU L-1 and 33, 73 IU L-1 of the separate compounds were seen in the essential oil. Besides, a huge elevation in serum LDH action was discovered in mice orally given the essential oil. The serum activity of important phosphatase was not significantly different (p £ 0.05) when compared to the control after the oil administration. Actually, all the properties acquired in these groups were within the normal range (Mitruka and Rawnsley In an investigation by Fayed , cypressup Fayed . Serum eThe function of renal tissues is obtained by screening for creatinine and urea. Since they remain overseen principally through filtration in the glomerulus with practically zero renal guideline or transformation throughout renal capacity deterioration Kamal . The masCupressus sempervirens leaf essential oil is toxic, and it is accounted here for the first time (Fayed Whenever there is elevation in the enzyme’s activities, there is also serious damage to the liver (Eteng et al. me Fayed .Cupressus sempervirens extracts has various biological and pharmacological effects. Recent pharmacological discoveries have shown the effects of the extracts in different points of view; these include anti-inflammatory, antioxidant, antidiabetic, antiviral, anti-malignancy and also their effect on cardiovascular and central nervous system. Conversely, these bioactivities originate from animal studies in vivo and in vitro studies though still inconsequential. It is interesting that in vitro bioactivity cannot fully elucidate the biologic effect in vivo, while the animal’s model is essential for pharmacological researches before clinical use. The bioactivities in vitro need to be explored by doing comprehensive research in future studies. Presently, the study done on the pharmacokinetics has recommended that the amentoflavone digestion was fast, and the bioavailability of bioflavonoid after indigestion orally is low in rats. Animal model experiments are few but available. We propose that the bioavailability can be improved by introducing synthetic compounds, modifying them structurally or adding specific drug that meets needs, which might be a focal point of the amentoflavones studies. Nevertheless, since there are some pharmacokinetic contrasts among normal and model animals of the evaluation of Cupressus sempervirens. Overall, the various bio and pharmacological activity of Cupressus sempervirens is owned to its rich phytochemicals which can be worked on for drug development in the future.From the previous studies, we could infer that


Omicron variant (B.1.1.529) is a dominant variant worldwide. However, the risk factors for Omicron variant clearance are yet unknown. The present study aimed to investigate the risk factors for early viral clearance of Omicron variant in patients with a history of inactivated vaccine injection.Demographic, clinical, and epidemiological data from 187 patients were collected retrospectively during the Omicron variant wave.+ T lymphocyte count were associated with SARS-CoV-2 RNA-positivity on day 7. Multivariable analysis showed that glucose ≥ 6.1 mmol/L and CD4+T lymphocytes count were independent risk factors for RNA positivity on day 7. 163/187 patients had an undetectable RNA test on day 14, and uric acid was the only independent risk factor for RNA positivity. Moreover, baseline glucose was negatively correlated with uric acid and CD4+ and CD8+ T cell count, while uric acid was positively correlated with CD4+ and CD8+ T cell count.73/187 and 114/187 patients were administered two and three doses of vaccine, respectively. The median duration of SARS-CoV-2 RNA positivity was 9 days, and the difference between patients with two and three vaccine injections was insignificant (P = 0.722). Fever was the most common symptom (125/187), and most patients (98.4%) had a fever for < 7 days. The RNA was undetectable in 65/187 patients on day 7. Univariable logistic analysis showed that baseline glucose, uric acid, lymphocytes count, platelet count, and CD4Omicron variant clearance was delayed in breakthrough cases with elevated fasting blood glucose, irrespective of the doses of inactivated vaccine. In the past 2 years, approximately 527 million patients have been infected by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and 6.3 million were deceased globally. Omicron variant B.1.1.529), first reported in November 2021, has become the dominant variant worldwide [.1.529, fThe spike receptor-binding domain (RBD) in the Omicron variant has enhanced binding properties with the human angiotensin-converting enzyme 2 (ACE2) receptor, which facilitates cell permeability . Since RBased on previous studies, blood glucose predicted the mortality of coronavirus disease 2019 (COVID-19), even in patients without a history of diabetes –16. HighTo date, the risk factors for Omicron variant clearance are yet unknown. Herein, we investigated the risk factors for 7- and 14-day viral clearance post-infection of Omicron variant in patients with a history of inactivated vaccine injection.A total of 239 patients with COVID-19 were admitted to the Third People’s Hospital of Changzhou between March 13, 2022 and May 10, 2022. All the patients were diagnosed according to the Chinese guidelines for the diagnosis and treatment of COVID-19 [ORF1ab and N genes < 35 indicated a positive RNA test. Duplicate tests were performed at 24 h if the result was negative (Ct ≥ 35).COVID-19 was confirmed based on the RT-PCR assay performed by the Laboratory of the Third People’s Hospital of Changzhou using a commercial kit . According to the manufacturer’s instructions, cycle threshold (Ct) values of All analyses were performed using SPSS 25.0 software . Data were expressed as median  or frequencies and compared using the Kruskal–Wallis test or chi-square test. Correlation analysis was performed using Spearman’s correlation test. Logistic regression analysis was conducted to identify the independent risk factors for viral clearance. A two-sided P-value < 0.05 was considered statistically significant.Among the 239 patients with COVID-19, 52 were excluded because of the undefined history of the vaccine. Data from 187 patients were analyzed, including 73 patients who had two doses of vaccine injection and 114 patients who received three doses. The duration from the first, second, and third vaccine injections to infection was defined as T1, T2, and T3, respectively. Only 5 patients had a T2 < 6 months, and none of the patients had a T3 < 1 month.For comorbidity, 3 patients had chronic hepatitis B, 8 had diabetes, 17 had hypertension, 1 had pulmonary tuberculosis, and 1 had chronic kidney disease. None of the patients was administered glucocorticoid or immunosuppressor at admission or during hospitalization.A total of 43 patients had elevated fasting blood glucose (≥ 6.1 mmol/L) at admission, among whom 7 patients had a diabetes history, and 1 patient was diagnosed with diabetes during hospitalization. Among the 35 patients who were not diagnosed with diabetes, the fasting blood glucose in 17 patients decreased to a normal level in a week without any hypoglycemic treatment, and 18 patients with slightly elevated glucose (≥ 6.1 mmol/L and < 7.5 mmol/L) did not undergo a repeat test. In addition, only 9 patients had a glucose ≥ 7.9 mmol/L at admission, and 4 patients had a normal glucose level in a week and were excluded from diabetes.Fever was the most common symptom , and most patients  had a fever duration of < 7 days. Other symptoms included cough , pharyngalgia , muscular soreness , headache , and diarrhea . According to CT scans, 13 individuals were diagnosed with unilateral pneumonia and 6 with bilateral pneumonia, while none developed severe pneumonia or ARDS. The median duration of SARS-CoV-2 RNA-positivity was 9 days, and the difference between patients with two and three doses of vaccine injection was insignificant .Z = 3.943, P < 0.01). In addition, neutrophils were higher, while the lymphocyte and platelet counts were lower in patients with a positive RNA test than those with a negative RNA test . Moreover, CD4+ T lymphocytes were higher in patients with viral clearance on day 7 .As shown in Table +T lymphocyte count are associated with SARS-CoV-2 RNA positivity on day 7. Then, multivariable analysis showed that baseline glucose and CD4+T lymphocytes count were independent risk factors for SARS-CoV-2 RNA positivity on day 7 (odds ratio (OR) = 0.631 and 1.001, 95% confidence interval (CI): 0.456–0.874 and 1.000–1.002, P < 0.01 and P = 0.02, respectively).As shown in Table 2 = 6.426, P = 0.01). Then, the multivariable analysis used glucose ≥ 6.1 mmol/L as a categorical value, and the data showed that glucose ≥ 6.1 mmol/L and CD4+ T lymphocytes count were independent risk factors for SARS-CoV-2 RNA-positivity on day 7 .More patients had a glucose level ≥ 6.1 mmol/L at admission in the RNA-positive group  compared to those in the RNA-negative group  . The CD4+ and CD8+ T cell count was high in patients with viral clearance .As shown in Table + and CD8+ T lymphocyte count were associated with SARS-CoV-2 RNA-positivity on day 14. Then, multivariate analysis showed that only baseline uric acid is the independent risk factor for SARS-CoV-2 RNA positivity on day 14 .As shown in Table + and CD8+ T lymphocyte count was deleted, and data showed that uric acid is the only independent risk factor for SARS-CoV-2 RNA positivity on day 14.Adjusted multivariable analysis was conducted when CD4+ T cell count, and CD8+ T cell count , while uric acid was positively correlated with CD4+ T cell and CD8+ T cell count (both P < 0.05). In addition, there was a positive correlation between CD4+ T and CD8+ T cell count .As shown in Fig. + T lymphocyte count were independent risk factors for SARS-CoV-2 RNA positivity. On day 14 after admission, SARS-CoV-2 RNA became undetectable in 87.2% of patients, and uric acid was the only independent risk factor for RNA-positivity.In the present study, we investigated the risk factors for early clearance of Omicron variant in patients with a history of inactivated vaccine injection. On day 7 after admission, 34.8% of patients had an undetectable RNA test, and baseline glucose ≥ 6.1 mmol/L and CD4The knowledge of prolonged SARS-CoV-2 shedding during Omicron waves is limited, especially for patients receiving inactivated vaccine injections. For most mAbs, the neutralizing activity against the Omicron variant is limited , 11, 24.+ T cell count is low in patients without early viral clearance and is negatively related to glucose. Thus, glucose and cellular immunity regulation may be an appropriate strategy for managing COVID-19.Several studies indicated that glucose at admission is an independent risk factor for the poor prognosis of COVID-19 during the Delta variant wave , 16, 20.+ and CD8+ T cells. Interestingly, the patients with lower serum uric acid levels experienced severe symptoms than those with higher uric acid levels [The present data showed that uric acid is the only independent predictor for SARS-CoV-2-positivity on day 14, negatively correlated with glucose, and positively correlated with CD4d levels , 28. SARd levels . The intd levels .The main limitation of the present study is that neutralizing antibodies at admission are not estimated. Regarding the Omicron variant spread, data from the present study highlighted the potential predictive value of metabolic factors, such as glucose and uric acid, in clinical practice. Typically, the benefits of the vaccine outweigh the potential risks , 32; henOmicron variant clearance was delayed in breakthrough cases with elevated fasting blood glucose, regardless of the inactivated vaccine doses.


The compounds were isolated from lyophilized extract of saffron using a Symmetry Prep C18 column (300 × 19 mm × 7 µm), and identified by spectroscopic techniques . The purity of crocins and picrocrocin was more than 97%. A novel method proposed to obtain working standards is simple and reproducible for the routine analysis of saffron quality control.Saffron is widely cultivated and used as a spice. Recently published data on the chemical composition and pharmacological potential of saffron determine its use in pharmacy and medicine. The proposed high-performance thin-layer chromatography (HPTLC) method allows good separation of 11 analytes. The saffron quality  assessment was based on the European Pharmacopoeia monograph and ISO 3632. The HPTLC method for the safranal, crocin, and picrocrocin quantification was proposed and validated. The crocins content in Ukrainian saffron was from 17.80% to 33.25%. Based on qualitative and quantitative assessment results, the saffron sample from Zaporizhzhia (Ukraine) had the highest compounds content and was chosen to obtain the working standards of picrocrocin and crocins ( The World Health Organization “Good Manufacturing Practice” (GMP) guidelines are rules that have been developed in accordance with modern pharmaceutical quality standards that establish requirements for the organization of production and the quality control of medicines.It specifically relates to the principles of quality assurance of pharmaceuticals and specifications of medicinal substances and dosage forms . These sCrocus sativus L. (Iridaceae) and is one of the oldest natural food supplements − and 365.2 [M − H + HCl]−  from saffron methanolic extracts, the UPLC-ESI-MS/MS method was used. The use of electrospray mass spectrometric (ESI-MS) detection in the negative ion mode makes it possible to obtain mass fragmentation of each compound as an additional method for confirming their structure . Their r + HCl]− . The abs44H64O24, m/z 976) produced the [M − H]− ion at m/z 975 at 5.64 min and exhibited absorption at maxima 261, 331, 440, and 467 nm. During the formation of adducts with formic acid in the mass spectrum of crocin 4, the peaks formed with m/z 1021.0 were identified as [M − H + HA]−, 978.3 [M − H + HCl]−, and 697.9 [M – H − Gnt + HA]−. The presented identification of mass ions is also characteristic of compounds containing one chlorine atom and is consistent with previously published observations of Lech et al. − ion at m/z 813. The UV-spectrum registered for crocin-3 exhibits absorption maxima at 261, 327, 440, and 466 nm.Crocin-3  crocetin esterified with one gentiobiose at one end and a free carboxylic group at the other showed the [M – H]– ion at m/z 651 at 7.29 min. The compound has similar ions fragmentation as a crocin-4 and crocin-3, and also has four maxima absorption at 258, 324, 435, and 460 nm characteristic of crocins-family. The obtained data were in a good agreement with previously published research [Crocin-2 (Cresearch . The preC. sativus stigma samples were harvested in 2019 in different Ukraine regions: Kherson (S1), Zaporizhzhia (S2), Odesa (S3), Chernihiv (S4), Mykolaiv (S5), and Vinnitsa (S6). The commercial saffron samples were purchased from Spain  (S7), Morocco  (S8), and Iran  (S9). Samples were identified by Dr. Mykhailenko (voucher specimens No. 20191–20198). All the numerical indicators of saffron samples quality were verified according to ISO/TS 3632 standards . For analysis were used: HPLC grade methanol, acetonitrile, and acetic acid (purity ≥ 98.0%) , and crocin reference standard with a purity ˃ 90% .Identification test. Saffron samples were ground using liquid nitrogen. Each powdered sample weighed 100 mg, and 50% methanol (10 mL) was used for extraction. The flask was placed in an ultrasonic bath  at room temperature (20 ± 2 °C) for 15 min. After extraction, extracts were centrifuged and the supernatant was used for chromatography (HPTLC).Isolation method. Saffron sample from Zaporizhzhia (Ukraine) was used for further analysis. The extract  of the chosen saffron sample was obtained similarly to the description above. Then, it was lyophilized at a reduced temperature of –25 °C and under reduced pressure overnight . The obtained lyophilized extract was used for preparative isolation of picrocrocin and crocin derivatives by HPLC and UPLC.® Linomat 5 autosampler, TLC scanner 3, Camag® TLC Visualizer. Separation was performed on TLC precoated silica gel 60F254 plates 10 × 10 cm . As a mobile phase was applied ethyl acetate: isopropanol: water (10:25:65 by v/v). Application conditions: the width—5 mm (5 μL), rate—50 nL/s. TLC plates were dried on the open air, then heated up on Camag HPTLC plate heater III at 60 °C for about 5 min. The scanning was performed at daylight, at 366 nm, at 254 nm (without treatment), as well as at 254 nm after plate derivatization with a spray reagent  and heating at 100–105 °C for 5–10 min. Quantitative analysis of the compounds was done by scanning the plates. The amount (%) of picrocrocin and safranal are presented in terms of crocin standard.For analysis, we used Camag TLC system: win CATS 1.2.2 software, Camagtrans-4GG, trans-2G, trans-3Gg).The Waters preparative HPLC Purification System  was used for preparative chromatography. The system comprised a 2767 Sample Manager, binary 1525 HPLC pumps, a 2488 UV detector and was controlled by Mass Lynx software version 4.1. The column isolation was a Symmetry Prep C18 (300 × 19 mm × 7 µm). HPLC conditions: mobile phase speed—17.0 mL/min; injection volume—1 mL; column temperature—20 °C; the mobile phase—a mixture of 0.1% acetic acid (A) and acetonitrile (B) with the linear gradient program 0–10 min, 5–15% B; 10–36 min, 15–20% B; 36–58 min, 20–40% B; 58–70 min, 40–50% B; 70–78 min, 50% B; 78–80 min, 50–95% B. The retention time of picrocrocin in these conditions was 45.73 min, for crocin—57 min, crocin (2)—60.4 min, crocin (3)—70.65 min. UPLC, HPLC, UV-vis, and MS studies were used for structural confirmation of picrocrocin and crocins  for three replications of each sample. Statistical analysis of HPTLC analysis was performed by one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison test with the software package Prism v.5.04 . The value of trans-4GG, trans-2G, trans-3Gg) is isolation and purification from saffron. The resulting compounds may be used as working standards for routine quality control. In addition, the proposed HPTLC method showed good separation of 11 tentative detected saffron compounds. A comparative analysis by the HPTLC method of nine different saffron samples showed a similar chemical composition and good quality of the samples. Moreover, the proposed method was used to determine the amount of main compounds  in Crocus stigma. Reference standard crocin showed good linearity of the calibration curves, low detection limit, and quantification limit. The obtained data show that the presented HPTLC method is simple, precise, accurate, and will be quite suitable for regular quality control of saffron in pharmaceutical analysis.This study suggests the simple, fast, selective, and environmentally friendly preparative chromatography method of picrocrocin and crocins (


Journal of Experimental Botany, Volume 72, Issue 17, 2 September 2021, Pages 6066–6075, https://doi.org/10.1093/jxb/erab278This is a correction to: Wasim A Iqbal, Isabel G Miller, Rebecca L Moore, Iain J Hope, Daniel Cowan-Turner, Maxim V Kapralov, Rubisco substitutions predicted to enhance crop performance through carbon uptake modelling, After being contacted by a reader, the authors have requested to amend some units and definitions in their manuscript and supplementary data file as follows:2 assimilation (µmol m−2 s−1)”.In Figure 3 and Supplementary Figure S1, y-axis units have been corrected to “Net CO2 assimilation rate (µmol m−2 s−1 days)”. Further explanation of the calculation of the values presented in Figure 4 and Supplementary Figure S2 has been added to the legends of these figures.In Figure 4 and Supplementary Figure S2, y-axis units have been corrected to “Seasonal net COIn Supplementary Table S1, the definitions of some of the parameters have been corrected. These definitions have been corrected throughout the supplementary data file.The version of record and supplementary data file have both been corrected online.


These distinct properties enable the construction of practical optoelectrical sensor arrays. However, contact engineering for MoS2 films is not still optimized. In this work, we inserted a graphene interlayer between the MoS2 films and Au contacts (graphene/Au) via the wet-transfer method to boost the device performance. Using graphene/Au contacts, outstanding electrical properties, namely field-effect mobility of 12.06 cm2/V∙s, on/off current ratio of 1.0 × 107, and responsivity of 610 A/W under illumination at 640 nm, were achieved. These favorable results were from the Fermi-level depinning effect induced by the graphene interlayer. Our results may help to construct large-area photonic sensor arrays based on 2D materials.Two-dimensional (2D) molybdenum disulfide (MoS Many stonducted ,5,6. Dasred SBHs . Howeverred SBHs ,8. To erred SBHs ,10. The contacts ,11,12. Ooperties ,11,13. Holiation ,15,16, a2 films grown by chemical vapor deposition (CVD) and Au contacts (graphene/Au contacts) to induce Fermi-level depinning leading to the enhancement in device performance of the resultant MoS2 device arrays. To investigate the effect of the graphene interlayer, we also compare the performance of conventional Au contacts in terms of electrical and photoelectrical properties.Here, we inserted a graphene interlayer between large-area MoS2 film via CVD using precursor powders, including MoO3  and sulfur . Each precursor was placed into two ceramic crucibles. A MoO3 crucible was placed at the center of a 1 inch quartz tube, and a sulfur crucible was positioned upstream of the MoO3 crucible. A Si/SiO2 substrate was loaded downstream of the MoO3 crucible. Then, the furnace was heated to 650 °C and maintained at this temperature for 40 min with a flow of Ar (50 sccm) under 0.3 Torr. After finishing the reaction, natural cooling was performed with 200 sccm of Ar flow. Graphene monolayer film was also synthesized on Cu foil  via identical CVD. The specific synthetic process is explained in our previous report [We first synthesized MoSs report .2 devices. MoS2 films were first prepared on Si/SiO2 substrate. To construct patterned graphene, first, photolithography and oxygen plasma etching treatment were performed. Once the patterned graphene was prepared, it was transferred onto MoS2 film via the wet-transfer method, where we used a buffered oxide etch solution to delaminate graphene. The second photolithography was carried out on patterned graphene electrodes, followed by metal deposition (50 nm-thick Au). To minimize the misalignment, the size of the Au electrodes was slightly bigger (200 × 200 μm) than that of graphene interlayers (150 × 150 μm). To prevent current leakage, channel definition was performed with a length of 14 μm and width of 40 μm using the third photolithography. Finally, the uncovered MoS2 area was etched using SF6 plasma treatment. We also fabricated MoS2 devices with Au-only contacts via the same procedure without a graphene layer to investigate the effect of the graphene interlayer.We used a standard photolithography method to fabricate MoS2 and graphene film grown by CVD. Transmission electron microscopy (TEM) measurement was performed to investigate the crystal structure of the obtained MoS2 film. Electrical and photoelectrical properties were characterized using a semiconducting parameter analyzer  with a 10−6 Torr vacuum at 80–300 K and light emitting diode (LED) visible light at 640 nm.Atomic force microscopy  and Raman/photoluminescence (PL) spectroscopy  equipped with a 532 nm laser were utilized at room temperature to explore the physical properties of MoS2 film synthesized via CVD. Overall, the obtained film shows good uniformity with no remarkable voids or cracks.  and on/off current ratio of 1.01 × 107(±2.52 × 106), compared with those of Au-contacted devices (mobility: 2.1 cm2/V∙s (±0.75) and on/off current ratio: 1.0 × 106 (±1.34 × 105)), as shown 2 devices with graphene/Au contacts while preserving off-state current levels. This enhancement presumably results from the decreased SBHs of the resultant devices and the Fermi-level depinning effect induced by the graphene interlayer, providing an efficient charge transfer between the graphene/Au contacts and MoS2 channels [Using the fabricated MoSeristics a,b. Notechannels ,21.T is the temperature (K), I0 is the saturated current level, q is the electronic charge, k is the Boltzmann constant, σ is the standard deviation of the Gaussian function of the SBH, A is the device area, A* is the Richardson constant, and Bφ is the barrier height. The Richardson constant is strongly associated with the effective mass of the material. It is estimated as A* = 4πqm*k2/h3, where m* is the effective mass of MoS2 and h is Planck’s constant [Rc) with respect to that of the Au-only–contacted devices I0T2)−q2σk2T2=ln, defined as R = Iph/(AP), where Iph is the difference between dark and photocurrents, A is the device area, and P is the light power density  than those of Au-contacted devices . This faster photosensor performance of the graphene/Au-contacted device is due to the efficient carrier separation behavior and low recombination rate caused by the improved carrier mobility [We finally assessed the photoelectrical properties of MoSt 640 nm a,b. Unde density c. Both d density . Furthermobility .2 device arrays with graphene/Au contacts. These graphene/Au-contacted devices have noticeable electrical performance, including a field-effect mobility of 12.06 cm2/V∙s and on/off current ratio of 1.0 × 107. This improvement can be attributable to the Fermi-level depinning effect caused by the graphene interlayer between Au and MoS2 channels, whereby the SBH and contact resistance are reduced. Furthermore, they show a visible photoresponse, achieving the responsivity of 610 A/W under illumination at 640 nm. This value is significantly comparable to that of TMDC photodetectors. Fabricating large-area MoS2 device arrays using graphene films constructed via CVD rather than exfoliated flakes offers insight into practical large-scale optoelectronic applications based on 2D TMDC materials.This report demonstrates the construction of large-area MoS


Nature Communications 10.1038/s41467-022-33362-1, published online 03 May 2009Correction to: 41; decadal solar irradiance42, mean annual insolation at 65°N48–50, and relative sea level47’ and now reads: ‘volcanic radiative forcing41; decadal total solar irradiance variations42 at earth surface /4, with mean albedo =0.29), mean annual insolation at 65°N48–50, and relative sea level47’. This has been corrected in both the PDF and HTML versions of the Article.The original version of this Article contained an error in Figs. 3 and 8, in which the global forcing curve was calculated and plotted incorrectly with incorrect description of variables in the Figure Caption of Figure 8. This read ‘volcanic radiative forcing