ILC2s interact with CD4+T cells to play a leading role in a variety of diseases through secretory factors. Here, we review the latest progress on ILC2s and CD4+T cells in the lung, the close relationship between the two, and their relevance in the lung disease and immunity. This literature review aids future research in pulmonary type 2 immune diseases and guides innovative treatment approaches for these diseases.Cytokine secretion, such as interleukin-4 (IL-4), IL-5, IL-9, IL-13, and amphiregulin (Areg), by type 2 innate lymphoid cells (ILC2s) is indispensable for homeostasis, remodeling/repairing tissue structure, inflammation, and tumor immunity. Often viewed as the innate cell surrogate of T helper type 2 (Th2) cells, ILC2s not only secrete the same type 2 cytokines, but are also inextricably related to CD4 The lung is a fragile organ, sensitive to temperature variation (cold and heat), constantly exposed to the external environment, and consequently susceptible to pathogen invasion. Thus, the lung is protected by a complex network of highly specialized immune cells and their mediators to support tissue homeostasis and prevent extensive tissue damage. The resting lung is prominently immune-tolerant, despite a plethora of leukocytes including B, T, and myeloid cells , which numbers and proportions changing dramatically during infection and inflammation.+T helper) 1, Th2, and Th17 T cell effector subsets, respectively [+T cells, especially their counterpart Th2 cells, share common stimulators and suppressors, as summarized in Recently a small subset of other immune cells was identified in the lung, namely, innate lymphoid cells (ILCs), which are distinct from T and B lymphocytes. ILC1s, ILC2s, and ILC3s act as the innate counterparts of the traditional Th CD4+T helperectively , but alsectively , 7. ILC2ectively  and thymectively , 9), andectively , 11, indectively , and TL1ectively , 14. In ectively  and Roraectively . Howeverectively . Thus, I+Th cells, ILCs also have a unique and nonredundant genetic organization in terms of kinetics, fine-tuning, and spatial organization of the immune response [+T cells in the lung and how their interaction participates in the immune regulation and lung homeostasis and will explore how their dysregulation can lead to lung diseases.Compared to CD4response . Here, win vitro and in vivo [–ILC2s and IL-10+Treg cells [in vivo, but can be restimulated upon new stimulus, similar to the Tl effector and memory cells [The ILC2 is a tissue-resident population and the major helper type ILCs in the steady-state lung of mice , 21. Dur in vivo . Increas in vivo . Some iI in vivo , suggest in vivo , 27. Ano in vivo . A new seg cells . This pory cells .Lung ILC2s are activated by the alarmins IL-33 , IL-25 , and TSL+ILC2s could lead to uncontrolled proliferation and activation of these cells [in vivo [γ [However, the inherent defect of programmed death 1(PD-1) signaling in KLRG1se cells . The inn[in vivo . Basophi[in vivo , and and[in vivo , endogen[in vivo , 45, pro[in vivo , and int vivo [γ  are key Interestingly, ILC2s are amateur antigen-presenting cells (APC) that can cooperate with dendritic cells (DCs) to maintain type 2 immune response . ILC2s nIn humans, ILC2s accumulate in fetal lungs at 10-fold higher levels than adult lungs . Another+T lymphocytes are a key element of adaptive immunity, and their role is to direct and enhance innate cell function [+T cells proliferate and differentiate into several possible effector subpopulations, including traditional T helper effector cells , T regulatory cells (Treg), and T follicular helper cells (Tfh) [+DCs have a self-maturation process, promoting Th17 differentiation at partial maturation and Th2 differentiation at maturation [CD4function . Naive Cls (Tfh)  that canls (Tfh) . Lung CDturation . These cα [α migrating upregulate T-bet transcription factor in CD11b+DCs inducing IL-12 secretion which in turn prevents the Th2 cells differentiation [Early Th2 cells tilt susceptibility to allergies and are often seen as remnants of fetal maternal symbiosis . Dopaminα . TNF-α mntiation . It has ntiation . Additiontiation .+T lymphocyte subsets (Th1/Th2 and Treg/Th17) [Fasting can inhibit the proliferation of Th2 cells in the lung and downregulate the level of Th2 cytokines . The pereg/Th17) .+Th cells while decreasing CD25+FOXP3+Treg. Additionally, fluoxetine promoted Th differentiation to Th1 cells while inhibiting Th2 and Th17 differentiation [Fluoxetine can inhibit cancer cell proliferation in cancer patients with depression . Fluoxetntiation . Major dntiation .+T cells, and crosstalk between the two contributes to their mutual maintenance, expansion, and cytokine production [There are two main mechanisms of crosstalk between ILC2s and CD4+T cells . During oduction , 76, as +T cells, particularly with Th2 cells, plays an important role in a variety of lung diseases  causing pathological changes . During in vivo required ILC2s activation, which was independent of ILC2s secreted cytokines but partially dependent on direct co-stimulatory interactions via ICOSL:ICOS [γ inhibits the activation of ILC2s and aggregation of Treg cells by IL-33 in infected tissues, and this inhibitory effect is enhanced with age and high-fat diet-induced obesity [Gastrointestinal helminth infection can cause local proliferation and accumulation of ILC2s in the lungs, which is promoted by IL-4/IL-13 secreted by Th2 cells . This ocOSL:ICOS . IFN-γ i obesity .+T cells and ILC2s cooperate to quickly expel worms within 48 hours [+T cells, so as to ensure the rapid activation and maintenance of IL-13-dependent M2 macrophages immunity in the lung [+T cells require further confirmation in human pulmonary parasitosis.During helminth infection, adventitial stromal cell depletion impairs the accumulation and function of lung ILC2s and Th2 cells partially dependent on adventitial stromal cells-derived IL-33, thus impairing the effect of expelling helminth . After h48 hours . This rethe lung . It shou+T cells in the lungs of mice [+T cells, but can also increase the cytokine production by CD4+T cells, which is dependent on direct contact between them [+T cells aggravate disease by promoting each other's proliferation and activation in mice.RSV infection is a major cause of bronchiolitis and pneumonia, especially in infants and young children . After t of mice . Adoptiveen them . NF-B/ILeen them , and NF-een them . Thus, iHuman primary bronchial epithelial cells infected with rhinovirus can induce IL-33, and the culture of human T cells and ILC2s with its supernatant can strongly induce type 2 cytokines . Viral i+T cells and ILC2s in the lung increases dramatically, with the significant increase of IL-33 expression in the lung [+T cells (rather than each cell population) can induce strong airway inflammation and antigen-specific type 2 immune response [Allergic airway diseases are characterized by sneezing, itching, wheezing, chest tightness, airway obstruction, and hyperresponsiveness . After athe lung . ILC2s aresponse . IL-5 anresponse . Thus, sresponse . In the response . IL-9 seresponse .+Th2 cells, which synergized with ILC2s, resulting in the deterioration of murine allergic airway diseases with prominent eosinophilia [Both T cells and ILC2s contribute to the deterioration of influenza-induced allergic airway inflammation, but the dynamics are different . Specifinophilia .+T cells, the effect of inhibiting both can be achieved, to reduce the pulmonary inflammation of asthmatic mice [Thus, by reducing the number of ILC2s or CD4tic mice , 103. Intic mice . The regtic mice . As an itic mice .+ILC2s are more resistant to this therapy in humans than Th2 cells and are closely related to asthma control status [+T cells and ILC2s [Similar findings have been found in human. A prospective study found that Th2 cells were high during asthma exacerbation and returned to baseline levels (similar to those of stable asthma patients) after aggressive treatment, suggesting that Th2 cells may be a biomarker of impending exacerbation in asthma patients . Pediatrl status . Interesnd ILC2s .+T was statistically significant in the stable state, in the acute exacerbation state, and after treatment, suggesting that Th2 cells can be used as a biomarker for acute exacerbations of asthma [However, the number of ILC2s was only found to be statistically significant in the acute exacerbation state versus the stable state, and not in the acute exacerbation phase versus after treatment in one clinical study . In thisf asthma . The inc+ILC2s peaked on day 7, and the expression of such cells was approximately five times higher than IL-13+CD4+T cells [+T cells [Pulmonary fibrosis is the end-stage change in a large group of lung diseases characterized by damaged alveolar structure, the massive proliferation of fibroblasts, and deposition of extracellular matrix , which m+T cells . This st+T cells . Areg pr+T cells , 114, an+T cells , which a+T cells .+CD4+T cells and regulating Th1/Th2 function can inhibit the progression of the COPD [COPD is an obstructive and progressive airway disease , which ithe COPD . The incthe COPD , 122. Ththe COPD . In addithe COPD .γ production by natural killer cells, but not by CD4+ and CD8+ T cells, thereby suppressing natural killer cell-mediated intrinsic antitumor immunity and leading to increased cancer lung metastasis and mortality [+T cells and CD8+T cells [Lung cancer is one of the most lethal cancers in the world . IL-33-dortality . ILC2s portality . They ac+T cells . IL-33 e+T cells .+/CD8+T cell ratios and a negative correlation with Treg cell levels [+T seems to promote the occurrence of human primary lung cancer and mouse primary and metastatic lung cancer and its role in human metastatic lung cancer required further research.In lung cancer patients, low levels of stimulator of interferon genes (STING) are strongly associated with poor patient prognosis . There wl levels . The lowl levels . ILC2s el levels –130. Thil levels . Anotherl levels . In summVaccines are biological products made from various types of pathogenic microorganisms for immunization . Vaccina+T in human vaccines is still lacking and urgently needs to be further studied.It was found that IL-33 secretion induced by epithelial cells necroptosis initiates APC- and ILC2-mediated T cell activation, which facilitated alum-enhanced Ag-specific IgA antibody production . IL-33-a+T cells are the command center of the adaptive immune system, also known as the “helper” of the immune system, and ILC2s are important innate lymphoid cells that serve as a bridge between innate and adaptive immunity. As summarized in +T cells interact in various ways, especially Th2 cells which seem to mutually promote each other's activation, proliferation, and function. Similar to the biological role of Th2 cells, ILC2s are involved in the defense against pulmonary parasitosis, the exacerbation of RSV infection, the aggravation of airway hyperresponsiveness during allergic diseases, the induction of lung injury, the acute exacerbation of COPD, the increase of cancer lung metastasis and mortality, and the improvement of vaccine efficacy, as summarized in CD4


High-performance blue-emitting counterparts are still eagerly required for next-generation full-color displays. However, it is challenging to obtain efficient blue perovskite NCs, especially in a deep blue region with an emission wavelength of around 460 nm or shorter. Herein, calcium halide and ammonium ions are applied simultaneously to modify the CsPb(Br/Cl)3 NCs in situ to reduce surface defects, finally remarkably enhancing the photoluminescence quantum yield (PLQY) from 13% to 93% with an emission peak at 455 nm and the Commission Internationale de l’Eclairage (CIE) coordinates at , which is close to the requirement of the Rec.2020 standard and also meets the requirement of blue emission in DCI-P3. Bright white emission and a wide color gamut are also achieved by combining the commercial red-emitting and green-emitting phosphors. The combination of time-resolved PL spectra and femtosecond transient absorption results discloses the reason for PLQY improvement as suppressing the nonradiative recombination.Colloidal cesium lead halide (CsPbX As emerging optoelectronic materials, colloidal cesium lead halide perovskite NCs show great potential as light-emitting diodes (LEDs). Considering their high photoluminescence quantum yield (PLQY), tunable emission wavelength covering the whole visible spectra, narrowband emission with high color purity, and solution processability ,3,4,5,6,2+) [2+ caused a dual-band emission. Additionally, copper [To figure out a solution for enhancing the performance of blue perovskite emitters, metal ion modification plays a critical role. As a special element that introduced an energy transfer pathway in the perovskite NCs, manganese ions (Mn2+)  were int, copper , tin [15, copper , zinc [1, copper  ions wer, copper  was used, copper ,23,24, a4+, exhibiting a similar ionic radius and chemical properties compared to alkali-earth metal ions [2 and power efficiency of 51 lm/W were obtained as well. The color gamut achieved by three primary color perovskite NCs exhibited excellent performance, with 147% DCI-P3 and 104.6% Rec.2020.Herein, we reported a facile, effective, and low-cost approach to achieving highly emissive blue-emitting cesium lead halide perovskite NCs by introducing calcium halide and ammonium ion additives. Calcium halide was used to passivate surface defects. Ammonium ions or NHtal ions ,26,27, c2CO3, 99.99% metals basis, Aladdin, Shanghai, China), ammonium carbonate ((NH4)2CO3, 99.999% metals basis, Aladdin), lead bromide , lead chloride , lead iodide , calcium bromide hydrate , calcium chloride , calcium iodide hydrate , oleic acid , oleylamine , 1-octadecene , and ethyl acetate  were used.Cesium carbonate .2, 0.752 mmol of PbCl2, i times of CaBr2·xH2O, and i times of CaCl2 were added, where i is the symbol of our sample that can be seen in the main part of our article, i = 0, 1, 2, and 3 (pristine sample corresponded to i = 0). Then, 6 mL of OA, 6 mL of OLA, and 4 mL of ODE were applied to dissolve these metal halide reactants and served as ligands and solvents. All reactants were heated at 120 °C under vacuum for 30 min until no solids could be observed in the flask. The temperature then rose to 170 °C under a nitrogen atmosphere and was maintained for 5 min for the complete dissolving of all reactants. Then, 2 mL of cesium and ammonium oleate solution which had been heated to 150 °C in advance was quickly injected into the lead, calcium, and halide reactants (precursor solution), and then the whole three-neck flask was put into ice water after 5 s reaction. The crude solution obtained was mixed with ethyl acetate with a volume ratio of 1:2 in a centrifuge tube. After a 10,000 rpm centrifugation for 5 min, the supernatants were discarded and solids were redispersed in 2 mL of hexane. Another low-speed centrifugation of 5000 rpm for 5 min was applied to remove large particles, and this time the supernatants were filtered by PTFE filters and stored for further use.To a three-neck flask, 0.752 mmol of PbBr3:Eu and 2SiO4:Eu, providing red and green emissions, respectively, and all three were mixed in a certain proportion. The others were consistent with the preparation method of blue light devices.The NCs solution was placed in a vacuum drying box at 60 °C to obtain a solid powder. The NCs solid powder was directly placed on the surface of the 395 nm commercial UV chip and encapsulated with a transparent resin protective cover to obtain a blue light-emitting device. The same quantity of each NCs solid powder was directly placed on the surface of the 395 nm commercial UV chip and encapsulated with a transparent resin protective cover to obtain a blue light-emitting device. The preparation of white photoluminescent devices was to mix blue perovskite NCs solid powder with two commercial phosphors, AlSiN3, a large number of halide vacancies, especially chloride vacancies [Generally, perovskite NCs synthesized by the two-precursor hot-injection method suffer from high-density surface defect states, especially blue-emitting perovskite NCs. Normally, the low performance of blue perovskite materials can be attributed to the following reason. Firstly, when the emission wavelength was tuned  to the bacancies ,30, exisacancies  where ex3 NCs were prepared via the hot-injection method proposed by Protesescu et al. [2) at the ends can effectively passivate surface defects and decrease non-radiative recombination, and protonated alkyl amines (–NH3+) with positive charges can fill cesium vacancies on the surface of nanocrystals. NH4+ is similar to the structure of –NH3+, and the effect of ammonium ions was speculated to reduce cesium vacancies. Indeed, the addition of ammonium oleate enhanced the luminescent property. The pristine synthesis without ammonium ions produced a PLQY of 5%, which was improved to ~12% by adding the ammonium oleate. However, the effect of ammonium ions was limited as the further increase in the number of ammonium ions did not show much higher PLQYs. Time-resolved PL decay measurements confirmed the limit effect of ammonium ions, as shown in Herein, the blue-emitting perovskite CsPb(Br/Cl)u et al. , with slu et al. ,32 have 2) and calcium chloride (CaCl2) to the reactants. Since calcium is an element with abundant storage in the earth shell, ranking 5th among all the known elements in the periodic table, we expected the introduction of calcium halide to be a low-cost strategy for highly emissive blue-emitting perovskite NCs. Here, we denoted Ca-i as the name of each sample, where i was defined as the multiple of calcium to lead used in the synthesis. With the purpose to determine the optimized value of i, three amounts of calcium were investigated, with i = 1, 2, and 3. The pristine sample with no calcium (the same amount of ammonium ions existed) was denoted as i = 0.Another method to realize further PLQY improvement was obtained by introducing calcium bromide  as well. We chose two commercial phosphors, AlSiN3:Eu and 2SiO4:Eu, to provide red and green emissions  and transmission electron microscopy (TEM) measurements were applied to explore the structural influence of calcium halide.3 (PDF#54-0752) and CsPbCl3 (PDF#18-0366) (XRD patterns exhibited distinct peaks located between the standard peaks of CsPbBr18-0366) a, confir18-0366) d,e. High18-0366) b,c revea18-0366) .2), protonated alkyl amines (–NH3+), and ammonium ions (NH4+), respectively  measurements were performed to characterize the composition of the perovskite NCs . The N 1ectively a.2 and CaCl2 solid powders, which shifted to lower binding energies. Zeng et al. investigated the XPS peaks of metal ions in different chemical environments, illustrating that metal-oleate exhibited a lower binding energy than that of metal-Br [7/2 and 142.98 eV for 4f5/2, and at 138.28 and 143.18 eV for the Ca-2 sample, respectively. The peaks in the Ca-2 sample revealed a 0.2 eV shift to higher binding energies compared to those in the Ca-0 sample, indicating that a stronger Pb–X  interaction was formed after introducing the calcium halide. The increase of atoms with high electronegativity around a given atom would cause the increase of positive charges on it [The detection of ammonium ions’ signals confirmed their successful incorporation. In metal-Br . Therefoes on it . Consequi from 0 to 3, reflecting better passivation of surface atoms and good dispersion in colloidal solutions, which was demonstrated by excellent configuration and no observed agglomeration in TEM images.Further exploration of the surface properties of our blue perovskite NCs was performed with quantitative XPS results. The molar ratio of halide to Pb a indicat2 bending vibration, and C–H stretching vibration peaks increased (The quantified Fourier transform infrared (FTIR) spectroscopy showed that with the increase of the Ca addition, the intensity of C=O stretching vibration, –NHncreased . The enhncreased , indicat1, and a larger one, t2, that were generally assigned [i = 0 to i = 2, with both radiative and nonradiative recombination lifetimes increasing. Meanwhile, the composition of nonradiative behaviors (A1) reduced and that of the radiative process (A2) increased, rendering highly bright blue emission , with a supreme PLQY of 93% and high color purity, with a narrow FWHM of around 20 nm. The modification successfully realized highly bright, deep blue-emitting perovskite emitters close to the requirement of the Rec.2020 standard and met the requirement of DCI-P3. Remarkable surface defect passivation was confirmed by time-resolved PL spectra, fs-TA, and XPS measurements. A white luminescent device near the standard white emission was fabricated with good performance. An extreme color gamut of 147% DCI-P3 and 104.6% Rec.2020 was realized by three primary color perovskite NCs, demonstrating the great potential of our modified perovskite NCs in the next-generation full-color display.


Myeloid sarcoma (MS) is a rare extramedullary mass with myeloid expression, which is easy to be missed and misdiagnosed, especially in the pediatric population. We analyze the clinicopathological characteristics, immunophenotypic, cytogenetic, and molecular studies, therapeutic approaches, and outcomes, to optimize the management of such patients.A retrospective, single-center, case series study of eleven children diagnosed with MS by pathology was performed.Class I-MS and four cases (36%) presented as Class II-MS. Immunohistochemically, MPO and CD117 were the most commonly expressed markers, followed by CD33, CD43, CD34, CD68, and lysozyme. Chromosomal abnormalities were detected in 4 patients. Two patients had the presence of deleterious mutations  on molecular detection. Ten patients were treated with chemotherapy based on AML regimens. The median follow-up time was 33.5 months in eleven patients. Two patients relapsed, one died, and one lost to follow-up. The 2-year overall survival (OS) rate estimated by Kaplan-Meier curves was 90.9% ± 8.7%, and the event-free survival (EFS) rate was 64.9% ± 16.7%.The male-to-female ratio was 8:3, and the median age at diagnosis was 7 years. The most commonly involved sites were the skin and orbital region, followed by lymph nodes, central nervous system, and testis. Seven cases (64%) with MS diagnosis is usually challenging. Adequate tumor biopsy and expanded immunohistochemistry are necessary for the correct diagnosis of MS. Early and regular systemic chemotherapy promises long-term survival. There aThe diagnosis of MS is based on clinical manifestations, imaging, mass biopsy, immunohistochemistry, cytogenetics, and molecular biology . There aHere, we report a comprehensive study of MS, including their clinical and laboratory manifestations, pathological and genetic features, treatments, and outcomes, in a series of 11 children. Literature review and single-center experience are used to improve pediatricians' understanding of the disease.This study was approved by the ethics committee of Xiangya Hospital Central South University in accordance with the Helsinki Declaration. Eleven children diagnosed with MS by pathology of the tumor mass were admitted from January 1, 2016, to April 1, 2022. Diagnoses were reconfirmed by two pathologists. The data we collected included the age, gender, clinical manifestations, site, history of AML or other neoplastic disorders, laboratory inspection, pathologic characteristics, radiological findings, treatment, and outcome.Class I, AML with MS; Class II, isolated MS ; Class III, extramedullary relapse of AML, including relapse after bone marrow transplantation; Class IV, acute/transformed stage of MDS/MPNs.WHO classifies them according to their extramedullary manifestations : Class IA. Morphological features: The morphology of MS manifests differently depending on the degree of myeloid differentiation, typically showing different stages of myeloid cell infiltration. B. Immunohistochemistry: it mostly showed MPO, CD43, CD34, CD13, CD33, CD117, CD14, CD11b, CD11c, CD68, CD163, lysozyme, and other myeloid and monocyte antigens positive, generally not expressing lymphocyte markers such as CD3, CD19, CD20)  panel including probes for deletion (5)/-5, deletion (7)/-7, t , t , and inversion (16), etc. histone lysine [K]-MethylTransferase 2A (KMT2A) gene formerly known as mixed-lineage leukemia (MLL) gene. Polymerase chain reaction (PCR) was used to detect fusion genes including MLL/AF10, AML1/ETO, PML/RARα, ET6V/RUNX1, BCR/ABL1, etc. Myeloid malignancies mutation panel detected in MS tumor mass or marrow by next-generation sequencing (NGS), such as fms like tyrosine kinase 3 (FLT3), tyrosine kinase receptor protein family KIT gene, nucleophosmin 1 (NPM1), CCAAT/enhancer binding protein alpha (CEBPA), additional sex combs-like 1 (ASXL1), etc.The G-banding technique was used to analyze the karyotype. Fluorescent Patients with MS were followed up through inpatient records, outpatient visits, and telephone calls, with a deadline of April 1, 2022. Overall survival (OS) was calculated as the time from diagnosis of MS until death or last follow-up. Event-free survival (EFS) was defined as the duration between diagnosis to the occurrence of events such as relapse, progression, death for any reason, or last follow-up.Descriptive statistics were performed for clinicopathological, molecular characteristics, and radiologic attributes. The quantitative data were represented by median , while the qualitative data were represented by a number of cases (%). OS and EFS were calculated with the Kaplan–Meier method.The major clinical features of all cases were summarized in All patients were evaluated by Computed Tomography (CT) or Magnetic Resonance Imaging (MRI) to identify the lesion site. Multi-site involvement in four patients (36%) and single-site in seven (64%). CT showed soft tissue density foci with uneven signal and irregular boundary in most patients. Bone necrosis was seen in some patients. MRI had no specific manifestations. T1-weighted images were mostly showed an equal or low signal, while T2-weighted mainly low signal, enhanced after contrast injection. Radiological data on part patients is shown in Class I-MS because of notable evidence of AML, with four AML-M2 (36%) and three AML-M5 (27%). Four patients (36%) presented as Class II-MS due to normal bone marrow, without a prior history of the hematopoietic disorder. No patients with Class III and IV-MS in our cohort. Patients diagnosed with AML underwent bone marrow flow cytometry. Six (55%) were positive for classic myeloid makers on flow cytometry, including MPO, CD33, CD34, and CD117. CSF flow cytometry in a patient with isolated MS showed positive myeloid markers for CD33, CD117, CD38, and CD64.Laboratory data is found in KMT2A (MLL) gene rearrangement, a common genetic abnormality of AML, were reported, and an expanded antibody panel retested IHC showed positive for CD33 and CD117 , FLT3 (p.I836del). The bone marrow myeloid mutation panel was identified positively in one patient with KIT (p.K558 > RGG), ASXL1 (p.S689fs*29), and DHX15 (p.R222G).Karyotyping results were available in eight patients because isolated MS was not routinely examined and one patient with AML refused. Cytogenetic abnormalities were found in four patients (36%) including +3, + 16, + 22, t , i (17) and del (1). Two patients with t   formed an KIT gene, and another patient with sorafenib for the mutation of the FLT3 gene. The median follow-up time was 33.5 months in eleven patients. two patients relapsed, one died, and one lost to follow-up. The 2-year OS rate estimated by Kaplan-Meier curves was 90.9% ± 8.7%, and the EFS rate was 64.9% ± 16.7%.Nine patients received surgical treatment. All patients were treated with chemotherapy based on AML regimens except one who eventually died of disease progression. AML regimens consisting of induction therapy with daunorubicin and cytarabine and consolidation therapy with high-dose cytarabine. CNS-MS patients were managed with central nervous system leukemia (CNSL) regimen. Eight patients (73%) achieved complete remission (CR) after the first or second induction chemotherapy. Of the eight patients, one patient discontinued chemotherapy and relapsed after 4 months w hile one lost to follow-up. Three patients who achieved CR to induction completed hematopoietic stem cell transplantation (HSCT). Of the three patients, two patients achieved sustained remission, and one relapsed 1 year after. One patient was treated with the addition of dasatinib due to the mutation of the In this study, we investigated a series of pediatric patients with MS at a chinese single institution. We highlights a precise pathological diagnosis based on immunohistochemical staining and the importance of genetic examination as well as early and regular systemic chemotherapy.According to the single-center data of 11 children diagnosed with MS, the most common subtype was MS accompanied by AML, which supported the previous literature –11. ApprRadiological test plays an important role in the early recognition of MS. CT and MRI imaging can assess the size and location of the tumor and help distinguish it from other lesions such as hematomas or abscesses . ConsistMS, especially isolated MS, is often a challenging diagnosis in pediatric patients . StudiesKMT2A (MLL) gene rearrangements or fusions, chromosome 5 or 7 monosomy, 5q-, and complex karyotypes of chromosomes  suggested a poor prognosis and high risk  gene were characterized by leukocytosis, CNS disease, and testicular involvement, which is consistent with the literature (Cytogenetic and molecular biological tests play an important role in the ancillary diagnosis, risk stratification and prognostic assessment of MS. Chromosomal abnormalities in 32%–84% of MS subjects , 21. Theigh risk –30. Patiterature , 28. NGSJohnston et al. showed that pediatric patients with orbital MS and CNS MS presented a lower relapse rate and better outcome than those without extramedullary manifestations (The prognostic data on the involved sites are different in the literature . Johnstostations . Studiesstations , 31. In stations . Hence, Based on previous reports, the treatment of MS is generally systemic chemotherapy, surgery, radiotherapy, HSCT, or a combination of these approaches. Surgery and radiotherapy cannot improve the overall survival of patients but are required when tumors cause obstruction or compression, or leukemia-related skin pain , 3, 33. In summary, MS show a variety of clinical manifestations that can be easily misdiagnosed. Adequate tumor biopsy and immunohistochemistry are necessary for the correct diagnosis of MS. Early and regular systemic chemotherapy promises long-term survival. Due to the rarity of MS, we reviewed too few patients to meaningfully study the impact of potential prognostic factors. It is hoped that future multicenter or even global collaborative large-sample prospective clinical trials will investigate pediatric MS in greater depth.


Melanogenesis is a major part of the environmental responses and tissue development of the integumentary system. The balance between reduction and oxidation (redox) governs pigmentary responses, for which coordination among epidermal resident cells is indispensable. Here, we review the current understanding of melanocyte biology with a particular focus on the “master regulator” of oxidative stress responses  and the autoimmune pigment disorder vitiligo. Our investigation revealed that the former is essential in pigmentogenesis, whereas the latter results from unbalanced redox homeostasis and/or defective intercellular communication in the interfollicular epidermis (IFE). Finally, we propose a model in which keratinocytes provide a “niche” for differentiated melanocytes and may “imprint” IFE pigmentation. Since t [LCEs]) ,9.Melanocytes originate from the neural crest and localize in the hair follicles (HFs) and IFE, endowing the body surface with pigmentation . Human m2, prostaglandin F2α, and nerve growth factor) and cell–cell interaction controls melanocyte proliferation, differentiation, melanogenesis, and dendritogenesis [The crosstalk between melanocytes and KCs through secreted factors , and the costimulatory receptor CD40 ,16,17,182− or H2O2 [Melanogenesis (melanin synthesis) is a complex biosynthetic process  that occ or H2O2 . Dopaqui or H2O2 . The rel2− or H2O2 from mitochondria [In general, the pheomelanin synthetic pathway requires cysteine/GSH, whereas the eumelanin pathway is expedited in the absence of thiols (high cystine/cysteine or GSSG/GSH ratios) ,28. Compchondria ,25, and chondria   mutation in the Slc7a11 gene reduces intracellular GSH levels and increases GSSG levels . The failure in response against extracellular oxidative signals skews melanogenesis toward the eumelanin synthesis pathway [NNT) controls melanosomal maturation (and eumelanogenesis) [+/NAD(P)H homeostasis by mediating electron transfer [Slc7a11 (sut mice) [NNT gene clearly differentiate skin pigmentary phenotypes, and inhibition takes place at the post-transcriptional levels [TYRP1 and TYRP2 [sut melanocytes, which mount suboptimal counter-responses against an extracellular oxidative milieu [A proper antioxidative response induced by changes in the intra-/extracellular redox milieu appears to give an important direction regarding the trajectory of the pigmentary pathways. Defective transmembrane cystine transport caused by the subtle gray ( pathway . The mitgenesis) . This intransfer . Althougtransfer , NNT inctransfer , similartransfer /Slc7a11 ut mice) . Howeverl levels . Existinnd TYRP2 . We shoue milieu , also exe milieu . Thus, re milieu . In summe milieu .Vitiligo is an acquired chronic pigmentary disorder that affects 0.5% to 2% of the world’s population without a clear preference for race or sex . The US Vitiligo pathogenesis involves multiple factors, such as genetic background, metabolic abnormalities, oxidative stress, generation of inflammatory mediators, autoimmune responses, and decreased melanocyte adhesiveness . These mHLA classes 1 and 2, PTPN22, IL2R α, GZMB, FOXP3, BACH2, CD80, and CCR6) and melanocyte-specific gene TYR in patients with vitiligo [NALP1 (the gene encoding NACHT leucine-rich repeat protein 1), a regulator of innate immunity, was found to be a risk factor for vitiligo [MTHFR (the gene encoding methylene tetrahydrofolate reductase), which regulates homocysteine levels, has been identified in patients with vitiligo [XBP1P1 (the gene encoding X-box binding protein 1) has also been associated with vitiligo. It is pivotal in attenuating the unfolded protein response and driving stress-induced inflammation in vivo [Multiple studies have revealed the genetic background of vitiligo development. Approximately 50 different genetic loci that contribute to the risk of vitiligo have been discovered, principally in European-derived whites and Chinese . A genomvitiligo . In addivitiligo . Recentlvitiligo . XBP1P1  in vivo .In vivo and in vitro investigations have confirmed widespread alterations of the antioxidant system in the skin and blood of patients with vitiligo ,61,62,632O2 induces vitiligo melanocyte cell death, oxeiptosis may contribute to vitiligo pathogenesis [Recently, a molecular mechanism involved in oxidative stress-induced melanocyte degeneration has been proposed. Oxeiptosis is an apoptosis-like, nonclassical, ROS-induced cell death pathway . Becauseogenesis . The micogenesis .Autoimmunity has been implicated in vitiligo pathogenesis . This is+ T cells that target melanocytes are responsible for melanocyte destruction [+ T cells produce several cytokines, including interferon (IFN)-γ [+ cytotoxic T cells (CTLs) to the skin [+ T cells and their localization within the epidermis [Oxidative stress connects innate immunity and adaptive immunity ,78. The truction . CD8+ T  (IFN)-γ ,89. IFN- (IFN)-γ . CXCL9 pthe skin ,91, wherpidermis ,91.2O2-induced oxidative stress [NRF2 or its downstream genes, such as NQO1 and PRDX6 reduces cultured melanocyte viability [sut mice that harbor the mutation in an NRF2-target Slc7a11 gene [sut mice exhibit reduced hair pheomelanin content, whereas that of eumelanin is barely affected although its effect on IFE pigmentation (tanning response) has not been assessed [2O2-induced oxidative stress promotes cytoplasmic translocation/release of high mobility group box 1 from NHEMs, resulting in indirect activation of NRF2 and its target genes  [NRF2 protects against cellular oxidative damage . This pre stress . α-MSH ie stress . NRF2 ace stress . Augmente stress . Accordie stress . Conversiability . Melanoca11 gene  exhibit a11 gene . Moreoveassessed . Furthernd GCLM) .2O2 levels [NRF2, NQO1, GCLC, and GCLM expression levels compared with the non-lesional epidermis [+ cytotoxic T cells (CTLs) [Evidence indicates that antioxidative response aberrations are central to vitiligo pathogenesis. SNPs in the NRF2 promoter may increase vitiligo risk , suggest2 levels ,61, and pidermis . An enhapidermis ,94, whicpidermis . Howevers (CTLs)  may brins (CTLs) , decreass (CTLs) . Collects (CTLs) . In thiss (CTLs) . These rs (CTLs)  as well s (CTLs) .N-acetylserotonin, and 5-methoxytryptamine) [Cistanche deserticola polysaccharides [Lycium barbarum polysaccharides [2O2, UVB, or MBEH. DMF is the methyl ester of fumaric acid and is one of the most successful NRF2 activators. DMF has been approved by the United States Food and Drug Administration (FDA) for relapsing-remitting multiple sclerosis, a demyelinating autoimmune disease [Augmenting the antioxidative responses of melanocytes and supporting their survival by pharmacological means could be attractive measures. Several compounds can protect melanocytes against oxidative stress through NRF2 activation . Melatonptamine) , vitaminptamine) , methylcptamine) , and vitptamine) ), naturaptamine) , ginsenoptamine) , Cistanccharides , glycyrrcharides , Lycium charides , paeonifcharides , 6-shogacharides , paeonolcharides , afzelincharides , apigenicharides , baicalecharides , vitexincharides , and bercharides ), and thcharides , dimethycharides , simvastcharides , moleculcharides , and colcharides ) are sho disease . The Eur disease . DMF tre disease . However disease . This mi disease ,78. TherVitiligo is not “just a cosmetic condition” but is psychologically devastating and stigmatizing . The psyThe aim of clinical management is to halt the autoimmune-driven depigmentation and restore the homeostatic pigmentation. Treatment options depend on several factors, such as disease subtype, extent, distribution, activity, patient age, phototype, effect on QOL, and motivation for treatment . These tPolypodium leucotomos, catalase/superoxide dismutase combination, and Ginkgo biloba may be administered with or without UV therapy [Although the abovementioned roles of oxidative stress in vitiligo rationalize antioxidant-based treatment, evidence of the efficacy of this treatment is quite limited . To achi therapy . Since t therapy . JAK inh therapy ,128,129. therapy  and 3 [1 therapy  trials; + cytotoxic epidermal resident CD8+ T cells efficiently [Keap1-null mutation not only augmented phase II detoxification but also led to uncontrolled keratinization of the squamous epithelium (SE) [Recent progress in vitiligo research has paved the way for disease pathway-based therapy. The IFN-γ-JAK-STAT pathway drives vitiligo pathogenesis, and JAK inhibitors, which presumably inhibit the effector function of CD49aiciently , hold priciently  appears Cutaneous pigmentary/depigmentary disorders, such as vitiligo or lentigines, do not necessarily accompany aberrant keratinization or acanthosis. Nonetheless, when considering the SE as a pigmentary “unit” , similar


Harumi tangor fruit with Ponkan as an interstock contains significantly higher levels of total soluble solids compared to Harumi tangor fruit cv.with no interstock. Transcriptome analysis of two graft combinations (Harumi/Hongjv (HP) and cv. cv.Harumi/Ponkan/Hongjv (HPP)) was conducted to identify the genes related to use of the Ponkan interstock. Soluble sugars and organic acids were also measured in the two graft combinations. The results showed that the contents of sucrose, glucose, and fructose were higher in the fruits of HPP than in those of HP; additionally, the titratable acid levels were lower in grafts with interstocks than in grafts without interstocks. Transcriptome analysis of HPP and HP citrus revealed that the interstock regulated auxin and ethylene signals, sugar and energy metabolism, and cell wall metabolism. Trend and Venn analyses suggested that genes related to carbohydrate-, energy-, and hormone-metabolic activities were more abundant in HPP plants than in HP plants during different periods. Moreover, weighted gene co-expression network analysis demonstrated that carbohydrates, hormones, cell wall, and transcription factors may be critical for interstock-mediated citrus fruit development and ripening. The contents of ethylene, auxin, cytokinin, transcription factors, starch, sucrose, glucose, fructose, and total sugar in HPP plants differed considerably than those in HP fruits. Interstocks may help to regulate the early ripening and quality of citrus fruit through the above-mentioned pathways. These findings provide information on the effects of interstock on plant growth and development. PBWRKY10 promotes the accumulation of organic acids in pears. The sugar-to-acid ratio is also an important determinant of fruit maturity  is a late-ripening cultivar that produces an oblate-shaped fruit; the rind color is orange and the fruit is easy to peel. The fruit ripens in January (C. reticulata × C. sinensis) is used as the base rootstock, using Ponkan (C. reticulata Ponkan) as an interstock can improve the fruit quality of ‘Harumi’ tangor. Thus, the main objective of this study was to explore how interstocks improve the quality of a common ‘Harumi’ tangor fruit. This effect was investigated using common ‘Harumi’ tangor grafted onto Hongjv  and cv.Ponkan/Hongjv .The mechanisms underlying interstock–scion interactions remain largely unknown. It is currently thought that the dwarfing mechanisms of rootstocks and interstocks alter the scion size, in which phytohormones may play important roles . Heterog January . This cu January . BecauseThis study was performed at the Citrus Modern Agricultural Park in Qingshen (29°86′–29°91′ N and 103°90′–106°38′ E) in Sichuan, China. The park has an elevation of 133 m, an annual accumulated temperature of >3,400°C, 3,000 h of sunshine, an annual frost-free period of 172–188 days, and a mean annual precipitation of 693 mm. Eighteen 2-year-old Hongjv seedlings were planted in September 2016. The first grafting (rootstock grafted with interstock) was conducted in early April 2017; Nine Hongjv were chip-budded with Ponkan at a height of 10 cm above the ground to form a Ponkan/Hongjv combination. Nine controls without grafting were also planted. The second branch grafting was conducted in March 2018, the interstocks were grafted with cultivar ‘Harumi’ tangor cv. cv.at 5 cm from the interstock/rootstock union to form a Harumi/Ponkan/red orange combination (HPP). Plants without interstock were also grafted with ‘Harumi’ tangor to form a Harumi/red orange combination (HP) at 10cm from the soil level. Plants were allowed to grow with full nutrition and standard treatments for citrus from 2018 to 2022.Fruit samples of ‘Harumi’ tangor were harvested at 45, 90, 135, and 180 days after flowering (DAF) from 2021 to 2022. Three biological replicates (three trees per replicate) were harvested at each developmental stage. The samples from the four stages were named as HP1, HP2, HP3, and HP4 for HP and HPP1, HPP2, HPP3, and HPP4 for HPP. Twenty-seven representative fruits (three fruits per tree) of each combination were sampled at each developmental stage. Fruits were transported to the laboratory on the day of harvest and then were divided into two groups. One group was used 15 samples for physiological index assessment, after measurement of TSS, single fruit weight, vertical diameter, and transverse diameter index, fruit flesh without skin from same batch of fruit were frozen in liquid nitrogen and stored at −80°C until high-performance liquid chromatography (HPLC) analysis. From the second group, the peel and flesh were isolated, and the flesh was frozen immediately in liquid nitrogen and stored at −80°C for subsequent analyses .A refractometer  was used for TSS measurements. The weight of each fruit was recorded using an electronic balance. The vertical and transverse diameters were measured for each fruit using a digital caliper gauge ( ± 0.01 mm).g for 10 min at 4°C. The residues from each treatment were extracted in triplicate, and the supernatant of each extracted residue was collected into a 10 mL volumetric flask and brought to a final volume (10 mL) with 80% ethanol. The samples were filtered through a membrane filter (0.45 μm pore size) before injection into the HPLC system. The filtered solution was used for sugar and organic acid analyses.Soluble sugars and organic acids were extracted as described by HPLC equipped with a refractive index detector . Samples were isolated on an Innoval NH2 column . Acetonitrile: water = 80:20 (v:v) was used as the mobile phase for isocratic elution with the following parameters: sample volume, 10 µL; flow rate, 1.0 mL·min-1; column temperature, 30°C; detection temperature, 40°C.The sugar content was analyzed using −1; column temperature, 25°C; detection temperature, 25°C. Organic acids were detected at a wavelength of 210 nm. For the determination of sugars and organic acids, three biological replicates were taken and technical replications were performed three times.Organic acids were analyzed using HPLC equipped with a UV detector . Samples were isolated on a C18-WP column . HPLC was performed using isocratic elution with the mobile phase . The parameters were as follows: sample volume, 10 µL; flow rate, 0.8 mL minand HPP across four developmental stages were harvested, and three biological replicates were harvested for each sample. A total of 24 transcriptome profiles were obtained by RNA-seq using the Illumina HiSeq™ 4000 sequencing platform at Novogene   .http://cucurbitgenomics.org/organism/21) using TopHat software version 2.0.12. Feature counting  was performed using HTSeq software version 0.6.1. Gene abundances were calculated and normalized to the number of reads per kilobase per million reads according to P-values < 0.05 were considered to reflect statistically significant differences and were adjusted using The raw sequencing data were deposited in the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus database under accession number PRJNA857756. The clean reads were aligned to the citrus reference genome , and first-strand cDNA was synthesized from total RNA using an RNA reverse transcription kit  according to the manufacturer’s instructions. To verify the accuracy of the transcriptome data, we used Primer 5 to design specific primers and performed quantitative reverse transcription-PCR analysis . One-way analysis of variance followed by Duncan’s multiple-range test was employed, and the standard deviation was calculated from three replicates. The differences between individual means were considered significant at Fruits of ‘Harumi’ tangor mandarins grown on HP and HPP trees were sampled at 45, 90, 135, and 180 DAF to explore the fruit quality at different stages of development  significantly differed during citrus fruit development  of fruit pulps harvested after 45 days were high in both the HP and HPP samples (2 (fold-change)| in expression of >0 and an adjusted P < 0.05. We identified 5,906 DEGs between the HP and HPP groups, including 1572, 441,1167, and 420 upregulated genes in the HPP1, HPP2, HPP3, and HPP4 groups, respectively, and 1422, 1207, 668, and 420 downregulated genes in the HPP1, HPP2, HPP3, and HPP 4 groups, respectively , glucan endo-1,3-β-glucosidase 14 (LOC18034912), TPS9 (LOC18044411), KOR1 (ciclev10003884m), BGL46 (LOC18052395), TPS11 (LOC18054628), XET 31 (LOC18034104), fructose-bisphosphate aldolase (FBA) genes , CSLG3 (LOC18046063), alkaline/neutral invertase CINVs , pyruvate kinase isozyme , inactive beta-amylase , and hexokinase . In contrast, the expression levels of several other genes were >2-fold higher in the HPP1 group, including genes encoding endoglucanases , PG , BGLU13 (LOC18044658), XYL4 (LOC18041989), glucose-6-phosphate isomerase , and malate dehydrogenase .The expression levels of genes related to carbohydrate metabolic processing were 1.3–10-fold higher in the HP1 group than in the HPP1 group. These upregulated genes included RPS30 (LOC18055660), RPL34A (LOC18048110), SUI1 (LOC18055413), alkaline/neutral invertase , phosphoenolpyruvate carboxykinase , cytochrome c , several genes encoding transcription factors (TFs) in the ERF family , and WRKYs . In contrast, the expression levels of several other genes were 1.2-fold higher in the HPP2 group, including genes encoding UTP-glucose-1-phosphate uridylyltransferase , 6-phosphofructokinase , sugar-phosphate translocator-related gene (LOC18047921), and pyruvate decarboxylase .The expression levels of genes related to organonitrogen compound biosynthesis were 2–60-fold higher in the HP2 group than in the HPP2 group. These upregulated genes included XTRs , XTHs , TCH4 (ciclev10010859m), phosphoenolpyruvate carboxylase , FBA (LOC18034756), pyruvate kinase , glyceraldehyde-3-phosphate dehydrogenase , NADH-quinone oxidoreductase , auxin-responsive protein SAUR , two ERF-type TFs , WRKYs , phosphoenolpyruvate carboxykinase , sucrose synthase , and cytochrome c .The expression levels of several genes encoding xyloglucan endotransglucosylase/hydrolase proteins were >1.5–11-fold higher in the HPP3 group than in the HP3 group. These genes encode FRK4, LOC18046804), and isocitrate dehydrogenase  were >1.2-fold higher in the HP4 group than in the HPP4 group. In contrast, the expression levels of genes encoding pyruvate kinase , cytochrome P450 , BR6OX1 (LOC18038016), CYP71B35 (LOC18043324), FAH1 (LOC18036838), CYP71B37 (LOC18047711), and CYP711A1 (LOC18054865) were 1.7–4.6-fold higher in the HPP4 group , serine carboxypeptidase-like , pyruvate kinase , fructokinase-4 (RPL18 (LOC18048753), and RPL36 (LOC18054512). However, the expression of genes encoding other ribosomal proteins were inhibited in the HPP2 group, including RPS30C (LOC18055660), RPL34A(LOC18048110), ARS27A(ciclev10017430m), RS21(LOC18041783), and a hypothetical protein (ciclev10003310m). Several genes encoding proteins important in plant hormone signaling and flavonoid biosynthesis were also downregulated by the interstocks in the HP3 group , XTH22 (LOC18054150), TCH4 (ciclev10010859m), IAA29 (LOC18054871) encoding the xyloglucan endo-transglucosylase/hydrolase protein and cytochrome P450 gene CYP75B1 (LOC18050323), flavonone isomerase gene TT5 (LOC18043493), and flavanone-3-hydroxylase gene F3H (LOC18036490). Genes encoding proteins involved in carbon metabolism and amino acid biosynthesis were also downregulated by the interstocks in the HP4 group, including L-3-cyanoalanine synthase gene CYSC1 (LOC18045998) and pyruvate kinase gene PK (LOC18046239).Ribosomes were affected by the interstocks , dihydrolipoyl dehydrogenase gene mtLPD1 (LOC18031585), and malate dehydrogenase gene MDH (LOC18045776) were upregulated in the HPP group at 45, 90, and 135 DAF and downregulated in the HPP group at 180 DAF. The isocitrate dehydrogenase gene IDH-III (LOC18037111), fructokinase gene FRK4 (LOC18046804), alkaline/neutral invertase gene CINV2 , hexokinase gene HXK1 (LOC18035909), and sucrase/ferredoxin-like gene (LOC18051223) were downregulated in the HPP group at 45, 90, and 180 DAF and upregulated in the HPP group at 135 DAF. The sucrose-phosphate synthase gene SPS3F (LOC18032032) was downregulated throughout all fruit-development stages in the HPP group. Two DEGs associated with the TCA cycle, BMY3 (LOC18032416) and DPE2 (LOC18039894), were downregulated throughout development and ripening in the HPP group, whereas the 1,4-alpha-glucan-branching enzyme gene SBE2.2 (LOC18041588) and pullulanase gene ATPU1 (LOC18043634) were downregulated in the HPP group at 45 DAF and upregulated in the HPP group at 90, 135, and 180 DAF.We analyzed the glycolysis pathway and associated DEGs (CSLG3 (LOC18046063) was downregulated during all stages. INVI/PMEIs (ciclev10000415m) and GH3 (LOC18035312) were downregulated in the HPP group at 45, 90, and 135 DAF and upregulated at 180 DAF. DHFR (LOC18039093), PAE8 (LOC18040742), and GH9B6 (LOC18033025) were upregulated in the HPP group at 45 DAF and downregulated at 90, 135, and 180 DAF. Additionally, one gene, GH28 (LOC18048144), was upregulated in the HPP group at all stages.DEGs related to cell wall metabolism are shown in DMR6, LOC18033472) and ethylene-responsive TF ERF104 (LOC18031355) were downregulated at 45 DAF and upregulated at 90, 135, and 180 DAF in the HPP group, whereas the gene encoding 2-oxoglutarate-dependent dioxygenase  was upregulated at 45 DAF and downregulated at 90, 135, and 180 DAF in the HPP group. Only ethylene-responsive TF 1B  was downregulated in the HPP group at all stages. Among the six DEGs associated with gibberellic acid synthesis, a synthetic degradation gene (GA20OX2) was downregulated in the HPP group at all stages. The auxin-related genes TIR1 (LOC18052163) and TSJT1 (LOC18054207) and cytokinin-related gene CKI1 (LOC18042595) were downregulated at 45, 90, and 135 DAF, whereas they were upregulated at 180 DAF in the HPP group , MYB, WRKY, bZIP, Aux/IAA, and C2H2 zinc finger proteins, as well as members of other major plant TF families , C2C2(Zn) DOF zinc finger family , MADS box TF family , MYB-related TFs , Kelch repeat-containing protein , and phosphatidylinositol N-acetylglucosaminyl transferase  were downregulated throughout development in the HPP group. Genes encoding Aux/IAA family , NIN-like bZIP-related family , nucleosome/chromatin assembly factor , MYB domain TF family , C2H2 zinc finger family , and growth-regulating factor  were upregulated throughout development in the HPP group  and glucan endo-1,3-β-glucosidase genes  were grouped into profile 0, whereas the MLO-like protein (MLO) gene was grouped into profiles 2, 9, and 10. Genes encoding ferredoxins  were associated with profile 2. Genes encoding galacturonosyl transferases  were grouped in profile 9. Genes encoding isocitrate dehydrogenase , 2-oxoglutarate dehydrogenase , phosphoenolpyruvate carboxylase , and succinate dehydrogenase  belonged to profile 10. Finally, genes encoding α,α-trehalose-phosphate synthase  were grouped in profile 19. Moreover, genes related to metabolic pathways, responses to oxidative stress, the biosynthesis of secondary metabolites, and plant hormone signal transduction were differentially expressed in the HP and HPP groups , cellulose synthase-like protein , and sucrose synthase . Additionally, genes associated with terms such as cellular lipid metabolic process, cell wall, multicellular organismal process, ADP binding, starch and sucrose metabolism, pyruvate metabolism, MAPK signaling pathway, and plant hormone signal transduction were enriched in the HPP group (A Venn diagram was used to display the number of DEGs at the four stages in the HP (1836) and HPP (1775) groups , ATP-dependent zinc metalloprotease , and acyltransferase-like protein (LOC18052762) were closely related to other genes , monogalactosyldiacylglycerol synthase , aldehyde dehydrogenase family 7 , and sugar transporter ERD6-like 6 (LOC18036954), as well as cinnamoyl-CoA reductase (LOC18041277), coumarin 8-geranyltransferase , and chloride channel protein  were also closely related to other genes , the sugar transporter ERD6-like 7 (LOC18046354), l-ascorbate peroxidase 3 , tryptophan aminotransferase-related protein 3 (LOC18043155), uridine nucleosidase 1 , tryptophan synthase α chain , and NADH dehydrogenase  were identified as hub genes in the tan-shaded module , protein aspartic protease in guard cell 1 , taxadien-5-α-ol O-acetyltransferase (LOC18044180), probable β-1,4-xylosyltransferase , auxin-responsive protein , a TF , serine carboxypeptidase-like 35 , and type-I inositol polyphosphate 5-phosphatase 4 .Among the significantly correlated modules, gene pairs with the top 20 weight values for each trait (high connectivity) were used to construct networks , synthetic degradation gene (GA20OX2), auxin-related genes TIR1 (LOC18052163) and TSJT1 (LOC18054207), and cytokinin-related gene CKI1 (LOC18042595) were downregulated in the HPP group. In many fruits, decreases in auxin levels occur during the initial stages of ripening  were downregulated in HPP citrus samples  were upregulated in HPP citrus samples  and energy metabolism  or those encoding TFs  were strongly affected by interstocks during fruit ripening. WGCNA revealed that genes related to the growth-regulating factor GRF10, sugar transporter ERD6-like, glycosyl transferase gene IRX9H, auxin gene IAA26, malate dehydrogenase gene MDH, and TF MYB16 were hub genes between the HP and HPP groups  can be found in the article/Project and experiment design, ZW and LL; experiment execution, LL, YL, XB, BX, XW, HD, and MZ; data analysis, ZJ, ZH and GS; writing, LL; review, BX and HD; project management, GS and ZW. All authors contributed to the article and approved the submitted version.This research was funded by the National Key R&D Program of the Ministry of Science and Technology project (2021YFD1600800) and Sichuan Science and Technology Department Projects (2021YJ0486).We thank all editors and reviewers for their critical reading of the manuscriptand their suggestions.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Nrf2 is a transcription factor encoding a large number of antioxidant enzymes. The activation of Nrf2 can have antioxidant and anti-inflammatory effects, which can also have complex crosstalk with NF-κB signaling pathway. The activation of Nrf2 can inhibit the M1 polarization and promote the M2 polarization through potential signaling transductions including TGF-β/SMAD, TLR/NF-κB, and JAK/STAT signaling pathways, with the regulation or cooperation of Notch, NLRP3, PI3K/Akt, and MAPK signaling. And the expression of heme oxygenase-1 (HO-1) and the negative regulation of Nrf2 for NF-κB can be the main mechanisms for promotion. Furthermore, the candidates of OA treatment by activating Nrf2 to promote M2 phenotype macrophages in OA are also reviewed in this work, such as itaconate and fumarate derivatives, curcumin, quercetin, melatonin, mesenchymal stem cells, and low-intensity pulsed ultrasound.Macrophages are the most abundant immune cells within the synovial joints, and also the main innate immune effector cells triggering the initial inflammatory responses in the pathological process of osteoarthritis (OA). The transition of synovial macrophages between pro-inflammatory and anti-inflammatory phenotypes can play a key role in building the intra-articular microenvironment. The pro-inflammatory cascade induced by TNF-α, IL-1β, and IL-6 is closely related to M1 macrophages, resulting in the production of pro-chondrolytic mediators. However, IL-10, IL1RA, CCL-18, IGF, and TGF are closely related to M2 macrophages, leading to the protection of cartilage and the promoted regeneration. The inhibition of NF-κB signaling pathway is central in OA treatment  In vivo imaging evidence in patients with OA indicates a crucial role for activated macrophages, which is linked to its severity. Moreover, the disruption of macrophage transition may contribute to chronic and irreversible inflammatory changes in OA-affected joints (Osteoarthritis (OA) is a highly prevalent musculoskeletal disorder characterized by pain, deformity, and functional deficits. Globally, it is a major medical and socioeconomic burden . Histolod joints . The actd joints .+ tissue-resident macrophages could restrict the inflammatory response  is a transcription factor expressed in most tissues and cells at a low level in the cytoplasm under homeostatic conditions through binding to Kelch-like ECH-associated protein 1 (KEAP1). Nrf2 is released in response to stress signals sensed by KEAP1, translocates to the nucleus, accumulates, and binds to antioxidant response elements (AREs) of target gene promoters . Finallyignaling . Moreoveignaling , 32. GSHhenotype . Howeverhenotype . Similarhenotype . Furtherhenotype . In summ2+ influx evoked by transient receptor potential vanilloid 1 (TRPV1) mediated inhibition of M1 macrophage polarization through the phosphorylation of calmodulin-dependent protein kinase II (CaMKII), while the specific inhibitor of Nrf2 counteracted the anti-inflammatory effect , has been found on OA chondrocytes and synovial fibroblasts . In factMMPs are a superfamily of proteases that can remodel and degrade ECM in connective tissues. By stimulating the release of MMP1, MMP3, and MMP13 in chondrocytes, TNF-α and IL-1β can affect the synthesis of proteoglycans, connexins, and type II collagen . MMP1 anThe failure in the appropriate proportion of M1 and M2 phenotypes can be the main cause of OA-related low-grade inflammation . After ain vitro, the cell death of chondrocytes, the release of glycosaminoglycans, NO, and molecules that promote ECM degradation and inhibit its syntheses, such as MMP3, MMP13, ADAMTS-4, and inducible nitric oxide synthase (iNOS) are significantly reduced. Moreover, the subsequent study confirmed that the ECM protective effects of IL-10 can be time-dependent  belonging to the growth factor family, of which the structure is related to insulin by 50% sequence homology , 73. IGFA models . FurtherA models . NotablyA models . ThroughA models –79. MoreA models . IGF-1 cA models . Further family , 84. SigThe conversion of the phenotype of macrophages in OA could play a role in treatment because the secreted cytokines regulate inflammation and cartilage metabolism Figure 1The polarization of macrophages dynamically adapts to changes in the microenvironment, and macrophage reprogramming has a complex mechanism. The most studied several pathways relative to reprogramming of macrophages include TGF-β/SMAD, TLR/NF-κB, and JAK/STAT signaling pathways, with the regulation or cooperation of Notch, NLRP3, PI3K/Akt, and MAPK signaling Figure 2via the phosphorylated type I receptor. As a result of the heteromeric trimer with SMAD4, activated R-SMADs can translocate to the nucleus and trigger transcription of target genes  are also the target genes of TGF-β, which regulates cell homeostasis . Of notetination , 94. In tination . The TGFtination .TLRs are prototype pattern-recognition receptors (PRRs) that recognize pathogen-associated molecular patterns (PAMPs) from microorganisms or DAMPs from damaged tissue . TLRs arThere are five TLR adaptors containing a Toll/IL-1 receptor (TIR) domain, among which MyD88 and TRIF bind directly to TLRs and recruit MAL and TRAM, respectively, while SARM negatively regulates these pathways. When the MyD88-dependent TLR4 signaling pathway mediated by TRAM is activated, TIR-domain-containing adaptor-inducing interferon-β (dependent on TRIF) can be recruited and then activates a cascade of proteins including TRAF6, which finally induces the degradation of IκBs, and the release of NF-κB (P50/65) and its translocation to the nucleus . NF-κB ivia the regulated transcription of NF-κB , which belong to the family of tyrosine kinases (TYKs). JAK1, JAK2, JAK3, and TYK2 have been indicated as JAKs, which can act as tyrosine kinase and bind tyrosine kinase-associated receptors intracellularly. Tyrosine kinase-associated receptors can bind various cytokines and growth factors, followed by the phosphorylation of these receptors and JAKs, and the initiation of the phosphorylation and activation of STATs  including STAT1, STAT2, STAT3, STAT4, STAT5A, STAT5B and STAT6 are latent in cytoplasm, and can be phosphorylated and activated of STATs –118.via JAK1 and Tyk2  signaling pathway, for example, GM-CSF-induced activation of JAK2 . PI3Ks aSTATs can also be activated by serine threonine kinases other than JAKs, such as extracellular signal-regulated kinase (ERK) . ERK belNrf2 belongs to the Cap’n’collar basic leucine zipper transcription factor family, within which the 605 amino acids act their roles as seven highly conserved functional NRF2-ECH homology (Neh) domains . It is tNext, from the N-terminal to the C-terminal, there are Neh4, Neh5, Neh7, Neh6, Neh1, and Neh3, respectively. Neh4, Neh5, and Neh3 are transactivation domains that mediate the interaction of Nrf2 with other coactivators. Neh4 and Neh5 can bind CBP/P300 , 152, whIn cells exposed to stress or electrophilic agents, the alignment of Nrf2 lysine residues is disrupted by the specific thiol residues, leading to the lysine residues being modified by electrophiles within the weak interaction with KEAP1, preventing ubiquitination, and ultimately releasing Nrf2 , 157. NrFurthermore, polarized macrophages can be used to target synovial inflammation caused by OA , 20. TheHeme oxygenase (HO) is a microsomal enzyme that degrades heme to carbon monoxide (CO), iron, and biliverdin, which plays a protective role in intracellular detoxification during tissue injury. HO regulates a range of anti-inflammatory, antioxidant, and anti-apoptotic pathways through heme degradation, and HO-1 is responsible for most intracellular detoxification among all the HO members , 160. ThBACH1 plays an important role in Nrf2/HO-1 transcriptional activity . BACH1 iThe promoter regions of HO-1 genes contain many AP-1 functional sites . AP-1 isvia anti-inflammation response and cell protection. HO-1 expression induced by sphingosine-1-phosphate in the supernatant derived from apoptotic cells has been indicated to play a vital role in M2 polarization of macrophages, via the STAT1/STAT3 heterodimer , hydroxyl radical, and singlet oxygen. ROS generated in cytoplasm and mitochondria act their key roles as signaling molecules to regulate physiological processes of macrophages. The ROS generated in cytoplasm and mitochondria serve as important signaling molecules to regulate macrophage physiological processes. NADPH oxidases (NOXs) transfer one electron from NADPH to oxygen to produce cytosolic ROS. Besides, the production of mitochondrial ROS is higher under the biochemical activities of ETC, monoamine oxidases, and P66shc , hydrogen peroxide  β resulting in the release of NF-κB in the pro-inflammation micro-environment. IKKβ can bind KEAP1 of NF-κB –205. On of NF-κB . NF-κB  is an esterified itaconate derivative that can be converted into itaconate by a direct modification of intracellular cysteines , 220. Diin vivo . In macrin vivo . Howeverin vivo .via decreasing oxidative stress and inflammation responses, and put a brake on the progress of OA in vivo  . CurcumiThe effects of curcumin in OA can also be independent of the Nrf2/HO-1 axis in reducing inflammation, preventing ECM degradation, and promoting cartilage synthesis , 233. Thvia promoting the degeneration of KEAP1 , is an indolamine with numerous functions in neural, endocrine, and immune physiological activities, playing a versatile role in the regulation of circadian rhythms, the defense against oxidative and inflammation, and the modulation of mitochondrial homeostasis. Mel is synthesized from tryptophan under 5-hydroxytryptamine in multiple extrapineal tissues , 252. In1 receptors can activate PI3K/Akt and ERK signaling pathways in synovial fibroblasts leading to the up-regulation of microRNA-185a, which can reverse OA-induced pathology in animal models through reducing the secretion of TNF-α, IL-8, and vascular endothelial growth factors  are heterogeneous stromal cells commonly sourced from adipose tissue, bone marrow and umbilical cord blood. MSCs have the capacity of differentiating into adipocytes, chondrocytes, and osteoblasts. Except for the capacity of multidirectional differentiation, MSCs exert broad immunoregulatory abilities, which can induce the specific polarization (M1 to M2) of macrophages to promote the repair of damaged tissues  actions –262. The actions .via M2 macrophage polarization. MSCs derived from bone marrow, which are labeled by iron oxide nanoparticles, can induce the increase in CD206-positive cells out of F4/80-positive macrophages, while can also induce the decrease in iNOS-positive cells out of F4/80-positive macrophages in animals experiencing OA due to destabilization of medical meniscus  with antioxidative characteristics nd TNF-α . Accordind TNF-α . For MSCnd TNF-α , 269. Threvealed . Besidesrevealed . Howeverammation . HoweverO-1 axis .2  outputs in a pulse wave mode of ultrasound with a non-thermal effect, at an intensity lower than 3 W/cm2 . The used TGF-β1 . LIPUS hd TGF-β1 .Besides, LIPUS can promote the cartilage differentiation of bone marrow-derived MSCs due to promoting the nuclear localization of SOX9 dependent on the phosphorylation of ERK1/2 . The comIn this review, we reviewed the role of macrophage phenotypes in driving and relieving inflammation due to OA and summarized how NF-κB, Nrf2, and their crosstalk shape macrophage polarization. As Nrf2 signaling is believed to impact cellular metabolism, studying the effects of activators of Nrf2 on macrophage metabolism and phenotype related to inflammation could reveal how OA can be treated through reprogramming macrophage functions. Furthermore, studying the effect of Nrf2 activation on macrophages in the OA microenvironment may suggest a potential anti-inflammatory therapy target. Therefore, it is imperative to identify the relationship between Nrf2, macrophage function, and the progression of inflammatory diseases.Conception and design of study, LW and CH. Drafting of article, LW. Revision of draft, CH. All authors contributed to the article and approved the submitted version.This work was supported by the National Natural Science Foundation of China (81972146 to CH), the Department of Science and Technology of Sichuan Province (2020YJ0210 and 2021YFS0004 to CH).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Erythrina velutina is a Brazilian native tree of the Caatinga (a unique semiarid biome). It is widely used in traditional medicine showing anti-inflammatory and central nervous system modulating activities. The species is a rich source of specialized metabolites, mostly alkaloids and flavonoids. To date, genomic information, biosynthesis, and regulation of flavonoids remain unknown in this woody plant. As part of a larger ongoing research goal to better understand specialized metabolism in plants inhabiting the harsh conditions of the Caatinga, the present study focused on this important class of bioactive phenolics. Leaves and seeds of plants growing in their natural habitat had their metabolic and proteomic profiles analyzed and integrated with transcriptome data. As a result, 96 metabolites (including 43 flavonoids) were annotated. Transcripts of the flavonoid pathway totaled 27, of which EvCHI, EvCHR, EvCHS, EvCYP75A and EvCYP75B1 were identified as putative main targets for modulating the accumulation of these metabolites. The highest correspondence of mRNA vs. protein was observed in the differentially expressed transcripts. In addition, 394 candidate transcripts encoding for transcription factors distributed among the bHLH, ERF, and MYB families were annotated. Based on interaction network analyses, several putative genes of the flavonoid pathway and transcription factors were related, particularly TFs of the MYB family. Expression patterns of transcripts involved in flavonoid biosynthesis and those involved in responses to biotic and abiotic stresses were discussed in detail. Overall, these findings provide a base for the understanding of molecular and metabolic responses in this medicinally important species. Moreover, the identification of key regulatory targets for future studies aiming at bioactive metabolite production will be facilitated. Erythrina velutina (Fabaceae) is a pioneer tree from the Caatinga, a unique tropical dry forest located in the semiarid region of North-eastern Brazil  are present in small amounts, which hampers extraction and detection by analytical techniques, and makes the process a major challenge for development of new products , and gel-free proteomics. This study provides new insights into the regulation and bioprospecting of specialized metabolites of Erythrina, serving as a reference platform for understanding and modulating the unique biochemistry of plants from this little-known peculiar and biodiverse semiarid biome.As part of our research efforts to dissect specialized metabolism in plants adapted to the harsh and particular conditions of the Caatinga, herein we applied integrated tools and carried out a comparative global analysis of the metabolites set present in Erythrina velutina Willd in different locations and subjected to a multi-omics analyses approach . A voucher specimen was deposited at the Herbarium of the Federal University of Rio Grande do Norte, Brazil, under the reference number UFRN16079. Authorization to harvest plant material was granted by SISBIO (327493) and access to the Brazilian genetic heritage by SISGEN (A8E4663). All samples were analyzed in four biological replicates.Harvesting and processing of plant material, metabolite extraction, and HR-LC-DAD-ESI-MS/AutoMS data acquisition were as described in Chacon et al. . Sampleshttp://mzmine.github.io/). Preprocessing was carried out for all samples in the following steps: mass calibration, mass detection (in both MS1 and MS2 levels), extraction of retention time (RT) information by chromatogram builder, peak deconvolution, mass grouping of isotopic patterns, and spectral alignment  and processed using MzMine2 (https://ccms-ucsd.github.io/GNPSDocumentation/) on the GNPS website (http://gnps.ucsd.edu). Moreover, to carry out spectral correspondence against libraries in the GNPS, the mass tolerance of precursor ions was defined as 2.0 Da, whereas ion tolerance of fragments of MS/MS was set at 0.5 Da. The library spectra were filtered in the same way as the input data. All matches held between network spectra and library spectra required a score above 0.7 and at least 6 shared peaks.Following preprocessing, the resulting data was inserted into the Global Natural Products Social Molecular Networking Platform (GNPS) . A minimum of 30% identity, 50% coverage and 10−10 E-value were used as parameters.Raw sequences were assembled using the software Trinity v2.11.0 . Initially, the database was downloaded, filtered for classification viridiplantae_odb10.2020.-09-10.tar.gz, confronted with the set of identified transcripts, followed by the execution of tBLASTN to predict the orthologous proteins.To assess the genetic completeness of the RNA-seq dataset, we used the BuscoV3 tool v4.1.4|2020-10-01 . After mapping, the GO terms and their respective p-values were inserted in the ReviGO tool v1.2 to detect the overrepresented terms, removing the obsolete and redundant GO terms using default criteria.Differential expression analysis was performed with the R limma package v3.48.3 , followed by BLASTp. Other statistics and functional analyses were performed using R v.3.5.2, including graphical representations.KofamKOALA was used to assign KEGG IDs to the translated transcripts , configured with a 3 cm x 100 μm trap column containing 5 μm, 120 Å C18 particles  connected in series to the 24 cm x 75 μm analytical column containing 3 μm, 120 Å C18 particles . The samples were injected to obtain 1 μg in the column, submitted to a linear elution gradient between solvents A (0.1% formic acid in water) and B (0.1% formic acid in acetonitrile) from 2% B to 35% B within 155 min.m/z between 300 and 1,650. The 20 most intense ions, above the intensity limit of 3,000 were fragmented, generating MS2 spectra, in the CID ion trap analyzer , configured to operate in DDA (data-dependent acquisition) mode, and the MS1 spectra were acquired in the orbitrap analyzer, with a resolution of 120,000 and a range of http://www.patternlabforproteomics.org/. The predicted protein sequences from the Erythrina velutina transcriptome dataset were used as the database and matched against raw files from the experimental proteome, including the 123 common contaminants from mass spectrometry. The Comet 2021.01 rev. 0 search engine was used for identifying the mass spectra    in PatternLab. Four biological replicates (two from the city of Acari and two from the city of Jardim do Seridó) were independently quantified in both leaves and seeds, with two technical replicates each. As stringency parameters, proteins with at least 2 unique peptides were selected, common contaminants and decoys were included in the search. We also performed the differential analysis of the abundance using PatternLab's TFold module to compare leaves and seeds. The statistical filters were Benjamin Hochberg q-value (FDR) 0.05 and F-stringency 0.13  (3702). After retrieval, the IDs were entered into the String v11.5 platform (https://string-db.org/), selecting the organism Arabidopsis thaliana. The network type was defined as a full STRING network and minimum required interaction score of 0.500 confidence.Initially, mapping of the ID codes of the differentially expressed transcription factors and transcripts of the flavonoid pathway in E. velutina, allowing the outline of their putative biosynthesis routes , were analyzed by LC-HRMS, in positive ionization mode. The MS/MS data, once converted in mzXML and processed in Mzmine2, was uploaded to the GNPS platform for spectral similarity calculation and molecular network , carbohydrates , chalcone, and flavonols (compound 11). In leaves, the major metabolites eluted between 6 and 10 min of analysis and were characterized as flavones , isoflavone (daidzein-8-C-glucoside), phenylpropanoid (compound 27), and amino acid (compound 38). Details of annotated metabolites can be found in Variability of the chemical profile in the different plant structures and samples was investigated to check for replicate consistency and part specific signatures. Chromatogram analyses revealed homogeneous profiles among the replicates and highly similar ones in apolar compounds presence  eluted in the beginning of the chromatographic run (RT <20 min). In this range, the numbers of metabolites distributed in leaves and seeds were similar, but with different proportions of metabolic classes between the plant structures. For example, the number of unique metabolites of seeds and leaves corresponded to 25 and 22 annotated compounds, respectively, and 20 were common to both plant parts. However, 64% of the leaf-exclusive metabolites were flavones and amino acids. In seeds, the exclusive presence of flavones, isoflavone, flavonols, carbohydrates, amino acids, and chalcone represented 68%. Among the metabolites present in both leaves and seeds about 70% were flavones and isoflavones.The 29 compounds that eluted at the final stage of the chromatographic run (RT > 20 min) were distributed in different proportions. In total, 18 metabolites were present in the two plant structures, whereas 8 and 3 exclusive metabolites were observed in leaves and seeds, respectively. The main metabolic class found after elution in this final stage corresponded to fatty acids, representing 100% of the seed exclusive metabolites , 50% of the leaf exclusive metabolites, along with glycolipids , and 61% of the metabolites common to both plant structures . Lignans, a terpene, coumarin, and triterpene saponin were also annotated.The annotation of metabolites, their interrelationship, and similarity analysis from large MS/MS data sets generates a vast amount of information from LC-MS/MS. Molecular network may be used as a tool to organize voluminous data sets and spot differences between groups  were annotated using the GNPS spectral libraries ,D. A totnd seeds .The 96 annotated metabolites were distributed in different classes, including primary and secondary metabolism consisting of 25 flavones, 16 fatty acids, 8 isoflavones, 6 flavonols, 6 amino acids and analogs, 5 coumarins and derivatives, 4 terpenes and derivatives, 4 carbohydrates, 3 anthocyanins, 2 saponins, 2 phenylpropanoid derivatives, 2 lignans, 2 aurones, 2 chalcones, 2 glycerolipids, 1 flavonone, and 6 other compounds .E. velutina showed some differences (C-glycosides) constituted the greatest share of compounds, representing 43%, followed by amino acids (compounds 14 and 16), terpenes (compounds 24 and 84), and fatty acids (compounds 79 and 95), with smaller proportions (~10%)  . In thisC-glucoside, diosmetin), isoflavones , and fatty acids .In contrast, flavonols  were the predominant seed-specific compounds, representing about 15% of the 28 metabolites annotated. Similar numbers of metabolites were found in the flavone pathway , coumarin and derivatives , carbohydrates , and fatty acids  (11% each). A flavonone that was found exclusively in seeds was annotated as dihydrotricetin. The compounds common to both plant structures seem to comprise mainly flavones , 80% of the flavones, 75% of the isoflavones, and 67% of the flavanols. This total corresponds to 40 glycosylated secondary metabolites, 27% present in both structures. Our previous work reported the great diversity and high number of transcripts that encode glycosyltransferases, many of which act in secondary metabolism, including flavonoid biosynthesis  and 30,218 transcripts in seeds with an average size of 1,203 bp ; in contrast, we obtained 17,822 transcripts with an average length of 1,504 bp contig in the joint assembly. The mean N50 value was 1,882 bp  tool was used to reconstruct a network of metabolic pathways. In total, 6,316 translated transcripts were annotated and assigned to at least 400 pathways. The category with the highest number of transcripts was biosynthesis of secondary metabolites (381 transcripts) , having p < 0.01, being 3,937 recorded with fold-change variation (log2FC) > 2 and < −2   . Next, wp-values were incorporated into the reviGO web platform. We obtained 108 and 19 non-redundant GO terms of metabolic processes and molecular functions in the up-regulated category, respectively. For the downregulated category, 277 and 79 non-redundant GO terms of metabolic process and molecular functions were recorded, respectively.After investigating representative subsets of major biological processes and molecular functions of regulated genes, we mapped 144 enriched GO terms to up-regulated genes , and 438In the biological process category  and moleIn line with the leaf physiological role in photosynthetic metabolism, enriched terms related to pigment biosynthetic process, carotenoid, isoprenoid, and chlorophyll were up-regulated in this organ, and down-regulated in seeds . A greatE. velutina subfamily Papilionoideae (Fabaceae), most studies are associated with the ability of this clade to synthesize isoflavonoids , EvHIDH (log2FC 4.19 and 6.13), EvDFR (log2FC 3.38), EvRhGT1 (log2FC 1.73) and EvCHI (log2FC 1.57 and 1.73) were up-regulated in seeds, whereas the transcripts EvCHI (log2FC−1.71), EvF3H-2 (log2FC−3.24), EvGT6 (log2FC−3.56), EvCHR (log2FC−5.30), EvCHS (log2FC−5.43), EvFNSII (log2FC−5.67), EvFLS (log2FC−7.37), EvOMT1 (log2FC−7.32) and a transcript Ev4CL (log2FC−8.00) were up-regulated in leaves , FBH4 (AT2G42280.1) and six differentially expressed, all down-regulated in seeds: two transcripts UNE12 (AT4G02590.2), one transcript PIF4 (AT2G43010.2), one transcription factor ILR3-like (AT5G54680.1), one transcription factor NAI1-like (XP_027907442.1) and a basic helix-loop-helix encoding gene . The AP2/ERF superfamily of transcription factors is the ERF family, which in turn can be divided into two essential subfamilies: ERF and CBF/DREB , MYB111 (AT5G49330.1) and MYB73 (AT4G37260.1). Only MYB111 was regulated showing a log2FC−3.52. However, it had an adjusted stimulus .In addition, candidate transcripts encoding for transcription factors distributed among the bHLH, ERF, and MYB families were examined for potential interactions with genes of the flavonoid pathway. Based on interaction network analyses, several putative genes of the flavonoid pathway and transcription factors were related, particularly TFs of the MYB family , 14.In total, 1,152 and 749 proteins were identified in leaves and seeds, respectively . After c(f). Two isoforms of 14-3-3-like protein were up-regulated in leaves (g1 and g2). The 14-3-3 proteins are known to function as phosphosensors in signal transduction of hormonal, environmental stimuli, and stress responses  and superoxide dismutase [Fe] (b)  were up-r = 0.352 for leaves and r = 0.144 for seeds)  were selected. Correlation analysis was performed in two steps . In the r seeds) ,I. In a r seeds) , 9. OverE. velutina chemistry, the metabolic profile was obtained from plants growing in their natural habitats, deep into the Brazilian Caatinga biome. As an initial strategy to gain insight on this subject, different plant parts (leaves and seeds) were examined. The comparison between the different plant parts showed leaves having a higher number of annotated compounds, in line with their active metabolism and photosynthetic capacity. Like many woody species of Fabaceae, mature seeds of E. velutina are of the orthodox type and display dormancy due to the presence of impermeable teguments , responsible for the generation of chalcone from p-coumaroyl-CoA and three malonyl-CoA molecules to form the two phenyl rings (rings A and B) of the flavonoid skeleton (C6-C3-C6). The assembly of the C ring is catalyzed by chalcone isomerase (EvCHI), generating naringenin, which eventually leads to flavonoid subclasses . Of the 3 DET's corresponding to EvCHI, one was 2 times more expressed in leaves, whereas the other two were up-regulated in seeds  and EvFNSII (down-regulated in seeds), two EvCYP75A6 transcripts and one EvCYP75B1 transcript (these three displaying no change). FNS is responsible for converting flavanones such as naringenin, eriodictyol and liquiritigenin into flavones, not only apigenin and luteolin, but also 7,4′-dihydroxyflavone  FNSII, classified as CYP93B16, directly converted a flavonone to 7,4′-dihydroxyflavone  and CYP75B (EvCYP75B1), shared high identity with two non-Fabaceae species, Campanula medium (65%) and Arabidopsis thaliana (99%), respectively. Based on in the Swiss-Prot database, their functional domain was confirmed in silico as being compatible with the P450 superfamily. The CYP75A (F3'5'H) and CYP75B (F3'H) subfamilies can catalyze the conversion of apigenin to luteolin (KEGG: map0094) and perform hydroxylation of the B ring of flavonoids, important for the biosynthesis of blue and red anthocyanins  and GT6 (EvGT6) (both down-regulated in seeds) would be involved. In this step, the flavonol kaempferol 3-O-glucoside was uniquely annotated in leaves of our metabolome. FLS is involved in the formation of kaempferol, which when overexpressed in Brassica napus, results in accumulation of this flavonol , together with OMTs and UGTs, would lead to the formation of the other flavonols identified exclusively in seeds or the formation of leucocyanidin (step 3).In the indirect path, there is initially the formation of dihydroquercetin, in which enzymes of the CYP75B superfamily participate, mainly those encoded by EvRhGT1 (up-regulated in seeds), EvGT5 (no change) and EvGT6 (down-regulated), in addition to EvOMT1, down-regulated in seeds, and showing corresponding lower abundance of its protein product. In a previous publication, we observed that leaves had more proteins with a functional UDP-glycosyltransferase domain than seeds; moreover, genes encoding enzymes putatively involved in the glycosylation of different flavonoids, including flavonols and anthocyanidins, were recorded  may play relevant roles during early germination, often in redox regulation  was expressed 10 times more in leaves compared to seeds. Auxin is a key regulator of plant growth and development, controlling gene expression through the ARF family (DNA-binding auxin response factors) , distributed in different families, were identified in Arabidopsis leaves, DEWAX2 is involved in downregulating cuticular wax biosynthesis, an important barrier protecting plants from abiotic and biotic stresses  and ethylene (ET), being also involved in growth and development processes mediated by gibberellins (GAs), cytokinins (CTK) and brassinosteroids (BRs)  - as one of the main regulators of the flavonoid pathway. These protein complexes can provide interesting models for analyzing environmentally controlled transcript regulation -responsive pathogen defense and wound response genes, in addition to its role in positively regulating flavonoid biosynthesis  - Finance Code 001).This work was supported by the Serrapilheira Institute (grant number Serra-1709-19691), by the Ministry of Science, Technology, Innovation and Communications - MCTIC, by CNPq/National Council of Science and Technology - INCT BioNat, [grant number 465637/2014-0], Sáo Paulo Research Foundation (FAPESP grant INCTBioNat 2014/50926-0 and 2020/02207-5), and by The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.