Given the controversial roles of FMS-like tyrosine kinase 3 inhibitors (FLT3i) in various treatment stages of acute myeloid leukemia (AML), this study was designed to assess this problem and further explored which FLT3i worked more effectively.FLT3(+) patients, or demonstrating the efficiency of allogeneic hematopoietic stem cell transplantation  in FLT3(+) AML. Relative risk (RR) with 95% confidence intervals (CI) was used for estimating complete remission (CR), early death and toxicity. Hazard ratio (HR) was used to assess overall survival (OS), event-free survival (EFS), relapse-free survival (RFS) and cumulative incidence of relapse (CIR).A systematic review, meta-analysis and network meta-analysis (NMA) were conducted by filtering PubMed, Embase, Cochrane library, and Chinese databases. We included studies comparing therapeutic effects between FLT3i and non-FLT3i group in AML, particularly p = 0.04) and refractory and relapsed FLT3(+) AML (rrAML)  compared to non-FLT3i arm, followed by improved survival  rrAML: OS, HR 0.60, p < 0.01; RFS, HR 0.40, p = 0.01). In addition, allo-HSCT improved survival in FLT3(+) AML , which was further prolonged by FLT3i administrated after allo-HSCT . Additionally, FLT3i consistently improved OS (p < 0.05) regardless of FLT3-ITD ratio, when compared to non-FLT3i group. Besides, FLT3i showed significantly increased risk of thrombocytopenia, neutropenia, anemia, skin- and cardiac-related adverse effects, increased alanine aminotransferase, and increased risk of cough and dyspnea (p < 0.05). In NMA, gilteritinib showed the highest probability for improved prognosis.After addressing all criteria, 39 studies were eventually analyzed. Better CR was accomplished by FLT3i in untreated AML  AML and further boosted survival benefits from allo-HSCT as maintenance therapy, suggesting better prognosis if FLT3i is combined before and after allo-HSCT. In NMA, gilteritinib potentially achieved the best prognosis, which should be identified in direct trials.FLT3i safely improved prognosis in induction/reinduction stage of  FLT3) mutations are detected in approximately one-third of patients (FLT3(+) patients with internal tandem duplication (FLT3-ITD) ranging from 3 to more than 100 amino acids located in the juxtamembrane region and a FLT3 point mutation in the tyrosine kinase domain (TKD) in approximately 8% of newly diagnosed AML  patients always manifest poor prognosis, distinguished by high resistance frequency to induction chemotherapy and relapse, decreased response to salvage therapy, and shorter survival. However, TKD influence on prognosis remains contradictory (Acute myeloid leukemia (AML) is a heterogeneous hematologic malignancy characterized by a maturation block and accumulation of myeloid progenitor cells . Among tpatients , comprispoptosis . FLT3-ITadictory .FLT3 inhibitors (FLT3i) are a type of tyrosine kinase inhibitors with FLT3 inhibitory activity that may be particularly utilized to treat FLT3(+) AML with likely improved prognosis. According to current clinical studies, they proved that AML patients benefited from FLT3i  patients were also summarized to observe the effects of FLT3i as the maintenance therapy after allo-HSCT. We further performed network meta-analyses (NMA) and ranked the prognostic effects of various FLT3i based on phase 2 and 3 RCT to check the most effective FLT3i.These FLT3i are used in various stages, including induction with/without consolidation therapy in newly diagnosed AML, maintenance treatment after allogeneic hematopoietic stem cell transplantation  and salvage therapy in refractory and relapsed AML (rrAML). Up to now, no extensive study has been found to comprehensively explore the role of FLT3i in various AML treatment stages and explore which FLT3i probably works the best. Herein, we conducted a meta-analysis in an attempt to clarify the clinical benefit and safety of FLT3i. Data from allo-HSCT in This study was conducted according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA)  and was FLT3”, “acute myeloid leukemia”, “AML”, “hematopoietic stem cell transplant”, “HSCT”, “sorafenib”, “lestaurtinib”, “midostaurin”, “quizartinib”, and “gilteritinib”. The included reports were: (i) published in English or Chinese, (ii) limited to retrospective cohort studies or RCT reporting the therapeutic effects of FLT3i on AML, especially for FLT3(+) patients, or restricted to studies demonstrating prognostic effects of allo-HSCT on FLT3(+) AML, (iii) designed to contain two arms or more for comparing prognostic influence of FLT3i or allo-HSCT with controls. Studies were excluded if they: (i) reported unavailable or insufficient data, (ii) were reviews, case reports, editorials and letters, (iii) had overlapping cohorts and (iv) were sing-arm studies.A literature search was conducted through databases of PubMed, Embase, Cochrane library, China National Knowledge Infrastructure, and Wanfang since inception until September 30th, 2020, following keywords “FMS-like tyrosine kinase 3”, “Study selection was conducted in two stages. Initially, abstracts and titles of potential literature were independently browsed and screened by QX and SH according to inclusion and exclusion criteria. Both reviewers then evaluated the candidate articles and decided on their inclusion. Any discrepancy was discussed and, if required, settled through discussion or consultation with a third reviewer (LY). After selecting the candidate studies, full texts were checked to identify final eligible ones.P-value < 0.05 implied publication bias existence.The methodologic quality of primary studies was assessed separately by two reviewers (QX and SH), based on Newcastle-Ottawa-Scale (NOS)  and CochClinical information from the included studies was extracted independently by two authors (QX and SH), and any reported disagreement was settled by discussion or consultation with the third author (LY). The extracted data were comprised of the first author, study characteristics, patients’ baseline and prognostic information. The endpoints included overall survival (OS), event-free survival (EFS), relapse-free survival (RFS), cumulative incidence of relapse (CIR), early death , adverse events and complete remission (CR), defined by revised International Working Group Criteria , withoutp-value of < 0.05. The χ2-based Q statistic was used to assess the heterogeneity among studies. Low, moderate, substantial and considerable heterogeneity showed 2I < 30%, 30%–50%, 50%–75% and > 75%, respectively. A P-value ≥ 0.10 implied no heterogeneity or slight heterogeneity, whereas P < 0.10 meant significant heterogeneity existence  for survival were calculated with the inverse variance method, and pooled RR and 95% CI for CR, early death and adverse events were produced from the Mantel-Haenszel method . Analysexistence . When P-via packages of “gemtc” and “rjags” in RCT. We also calculated HR or risk ratios (RR) regarding non-FLT3i group as the baseline to act as the effect measure, displayed in forest plots, where RR and HR with 95% credible intervals (95% Crl) were utilized to explain the extent of effects in CR and survival, respectively. The range of 95% Crl without covering 1.00 implied statistical significance. To estimate relative HR and RR, a Markov Chain Monte Carlo simulation was finished with 10000 adaptations and 100000 iterations of each of the three automatically generated Markov chains. After finishing all simulations, NMA determined the probability that each treatment would be best by calculating the probability of simulations in which a certain treatment ranked best. For each iteration, therapies were ranked according to the evaluated log HR or log RR. The data from Bayesian NMA were compared with data from pairwise meta-analyses to assess inconsistency using the node splitting method  AML, except for Roellig et al. (FLT3(+) AML patients given sorafenib-related therapies as induction strategy, with or without consolidation and maintenance therapy following allo-HSCT. Six studies (n = 951) only focused on the maintenance therapy of sorafenib after allo-HSCT, and two studies (n = 235) regarded sorafenib as part of salvage therapy following relapse. Midostaurin was used in the stages of induction, consolidation and maintenance treatment in CR in four studies (n = 1604) and one article (n = 60) reported results of midostaurin-related maintenance therapy after allo-HSCT. For lestaurtinib studies, two (n = 500) focused on the induction stage in untreated FLT3(+) AML and one (n = 224) involved salvage therapy in FLT3(+) rrAML. In addition, the clinical efficacy of gilteritinib (n = 371) and quizartinib (n = 367) was evaluated in RCT for rrAML, and two studies (n = 178) combined several FLT3i. Finally, fourteen studies (n = 1797) compared prognostic discordance between allo-HSCT and non-HSCT in FLT3(+) AML.Totally, 6859 AML patients were included, and their studies were conducted in g et al.  and Servg et al.   in FLT3i groups but inconsistently in prolonged EFS  and better RFS   and better RFS    and Xuan= 0.025) , the comp = 0.01; heterogeneity: 2I = 13.3%, P = 0.330) and RFS . Besides, midostaurin consistently prolonged OS  but lestaurtinib did not provide better survival (p > 0.05).The subgroup analyses were further evaluated among various FLT3i (FLT3(+) AML were then investigated in 14 studies . OS and RFS were improved by allo-HSCT but with high heterogeneity . After sensitivity analyses, combined HR of OS and RFS became consistent, demonstrating pooled HR of 0.53 for OS  and pooled HR of 0.57 for RFS , respectively .We next analyzed FLT3i role as the maintenance treatment after allo-HSCT in eight studies, consistently showing better survival , and only one study was involved in gilteritinib, quizartinib, and lestaurtinib, respectively. Better OS was observed in gilteritinib  and quizartinib . However, lestaurtinib did not support better survival (p > 0.05).Prognostic effects of FLT3i in salvage regimen in rrAML were estimated among seven studies. After excluding Levis et al.  to decrep = 0.04; heterogeneity: 2I = 63.9%, P = 0.005) and rrAML . After sensitive analysis, combined RR of CR in rrAML became consistent, demonstrating pooled RR of 0.61 . In the subgroup analyses for newly diagnosed AML , 0.85  and 1.36 , respectively. In the subgroup analyses for salvage regimen in rrAML (p = 0.01), 0.79 , 0.45 , and 0.56 , respectively.CR from 15 studies was assessed  and low ratio  when compared to non-FLT3i group. However, FLT3i significantly prolonged RFS in patients with high-ratio FLT3-ITD  rather than those with low-ratio FLT3-ITD mutations , when compared to non-FLT3i arm. Finally, there was only one study involved in EFS, showing no prolonged survival in FLT3i group in both of high and low FLT3-ITD ratio.The treatment effectiveness of FLT3i was then analyzed in patients with low and high vs. non-FLT3i: 28.96% vs. 18.36%; RR, 2.20; 95% CI, 1.39–3.48, p = 0.0008; 2I = 55.2%, P = 0.108), neutropenia , as well as anemia  regardless of grades.Totally, early death and 21 types of toxic effects were analyzed , especially for grades ≥ 3  with low heterogeneity . FLT3i also consistently showed a tendency of increased risk for all-grade gastrointestinal-related toxicities . Additionally, significantly increased alanine aminotransferase was found in FLT3i group at all grades  and grade ≥ 3 . Cardiac-related adverse events were mainly reported in sorafenib in all grades  and grade ≥ 3 . Higher risk of all-grade cough  and dyspnea  was reported in FLT3i group.Besides, FLT3i were clearly related to higher risk of skin-related adverse effects at all grades .P < 0.10) existed. The heterogeneity sources were listed in FLT3(+) AML, as well as OS of FLT3i in salvage regimen. The funnel plots were illustrated in Sensitivity analyses were conducted if high heterogeneity  and OS   and ServFLT3(+) AML, and which FLT3i might be the best, which were investigated in this study. FLT3i possibly supported better CR in induction and salvage therapy that further boosted the improved survival of FLT3i. Allo-HSCT improved survival in FLT3(+) AML and FLT3i as the maintenance therapy after allo-HSCT might further enhance survival benefit gained from allo-HSCT. FLT3i also improved OS regardless of stratification of FLT3-ITD ratio, when compared to non-FLT3i group. Additionally, FLT3i consistently showed significantly increased risk of thrombocytopenia, neutropenia, anemia, skin-related adverse effects, increased alanine aminotransferase, cardiac-related adverse events, cough and dyspnea, but were not associated with high early death and increased risk of GVHD. NMA showed that gilteritinib probably favored the highest possibility toward better prognosis, which should be identified in more direct head-to-head RCT.Until now, no consensus was present on FLT3i role in various AML treatment stages, especially for FLT3 was the antileukemic activity of sorafenib in inhibiting other kinases such as RAF  (FLT3(+) patients aged 18-59 years, midostaurin plus chemotherapy achieved prolonged survival, regardless of FLT3-TKD and different levels of mutant FLT3-ITD ratio, probably resulting from enough exposure to this inhibitor. Given a benefit among patients with low allelic FLT3-ITD mutation and a large disease burden, the benefit of this multitargeted kinase inhibitor might lie beyond its ability to inhibit FLT3, like inhibiting KIT in FLT3 (–) AML (FLT3(+) AML, possibly due to the rising level of FLT3 ligand induced by chemotherapy, which could interfere with activity of FLT3 inhibition, including lestaurtinib  AML  confirmeaurtinib .FLT3(+) AML. However, some variables greatly affect the effectiveness of allo-HSCT, including disease status (CR or not), FLT3 variables , and using FLT3i before and/or after allo-HSCT. Unfortunately, no RCT assessed the most suitable post-remission treatment in FLT3(+) AML, considering diverse combinations (FLT3(+) AML (30%-59%) (FLT3-ITD(+) patients undergoing allo‐HSCT (FLT3-ITD(+) AML. In the RADIUS phase 2 RCT (FLT3(+) AML patients may still benefit from sorafenib after allo-HSCT because of the less use of other FLT3i  . Indeed,30%-59%)  showed ello‐HSCT , probabise 2 RCT , midostaer FLT3i .FLT3(+) AML (FLT3(+) rrAML, we also explored consistent or different therapeutic efficiency between various FLT3i in such patients. Except for lestaurtinib, the combined OS could be consistently improved by sorafenib, gilteritinib, and quizartinib, showing the potential of FLT3i in treating FLT3(+) rrAML. A phase 3 RCT randomized 371 FLT3(+) rrAML to either gilteritinib or salvage chemotherapy (FLT3(+) rrAML patients also confirmed quizartinib availability in improving prognosis (p < 0.05). Until now, the promising efficiency of gilteritinib and quizartinib could be observed in FLT3(+) rrAML in the two large RCT. However, a lack of evidence is present to add the two FLT3i to induction, consolidation, and maintenance therapy after allo-HSCT. Several phase 2 or 3 RCT are being processed . Additionally, two retrospective studies of sorafenib from Bazarbachi et al. (FLT3(+) rrAML, which should be further confirmed in RCT. Finally, midostaurin might be specifically effective in the untreated AML rather than rrAML, since in vitro studies, midostaurin had broader activity and might achieve greater clinical utility in newly diagnosed AML with blasts tending to be less addicted to FLT3-mediated signaling than rrAML (Due to high refractory and relapse incidence in 3(+) AML  and limiotherapy , showingi et al.  and Xuani et al.  also revan rrAML .FLT3-ITD(+) AML stratified by ITD allelic ratios, showing that FLT3i consistently achieved significantly improved OS in both of high and low ratio crossing sorafenib, quizartinib, midostaurin and even lestaurtinib, further suggesting the benefits of FLT3i in FLT3-ITD(+) AML. However, there was no information involved in assessing clinical efficiency of gilteritinib in patients with different levels of FLT3-ITD ratio, which needs to be further explored in more studies.We also explored the effectiveness of FLT3i on For adverse events, FLT3i were significantly associated with increased risk of thrombocytopenia, neutropenia and anemia regardless of grades, high risk of skin- and cardiac-related adverse effects, especially for grades ≥ 3, all-grade increased alanine aminotransferase, high risk of all-grade cough and dyspnea, as well as a tendency of increased risk for all-grade gastrointestinal-related toxicities. However, these adverse events were generally manageable based on treatment interruptions or dose reductions . MoreoveFinally, due to co-existence of several FLT3i, it would also be meaningful to explore which inhibitor might be the best. Herein, we finished an NMA based on all RCT to settle this problem. The results displayed that gilteritinib probably tended to favor the highest probability of improving prognosis compared with other FLT3i, standard of care and standard chemotherapy. The relevant RCT involved in gilteritinib for rrAML treatment also illustrated improved survival as mentioned above . HoweverFLT3(+) AML and therapeutic efficiency of FLT3i as maintenance therapy after allo-HSCT and as salvage regimens in rrAML were summarized and explored, showing that combining FLT3i, especially sorafenib, into the treatment before and after allo-HSCT might be more beneficial in improving prognosis, which should be further explored in RCT. Our study also primitively proposed NMA to compare various FLT3i, but with limitations as mentioned above. There were also several limitations in this study. Of 39 studies, 28 studies were retrospective, making it difficult to precisely control selection, attrition, information and confounding bias. In particular, due to RCT lack, limited results and conclusion were obtained from NMA. Besides, some data were extracted from Kaplan-Meier survival curves and numeric reports, probably resulting in slight disparity with the fact. Finally, this study did not make subgroup analyses based on age, co-existing mutations with FLT3 mutations, and cytogenetic stratifications due to limited relevant data in primary studies. As a consequence, we will continue updating this study to explore suitable treatment of FLT3i in AML patients.Overall, our study was up to now the biggest and the most comprehensive meta-analysis involved in various FLT3i in AML, containing all RCT and retrospective cohort studies. In addition to FLT3i function in induction stage in untreated AML, the role of allo-HSCT in FLT3(+) AML and salvage therapy of FLT3(+) rrAML and further enhanced survival advantages from allo-HSCT as the maintenance therapy. This probably indicates that better prognosis could be achieved if FLT3i is added into both treatments before and after allo-HSCT. Besides, FLT3i probably improved OS regardless of FLT3-ITD ratio. Additionally, FLT3i were significantly linked to increased risk of all-grade thrombocytopenia, neutropenia, anemia, all-grade skin- and cardiac-related adverse effects, especially for grade ≥ 3, as well as all-grade increased alanine aminotransferase, enhanced risk of all-grade cough and dyspnea, and a tendency of increased risk for all-grade gastrointestinal-related toxicities, but not related to higher incidence of early death and GVHD compared to non-FLT3i group. In NMA, gilteritinib potentially accomplished the best prognosis, which should be identified in direct head-to-head RCT.In this work, FLT3i reinforced better prognosis in the induction stage of newly diagnosed The original contributions presented in the study are included in the article/QX and LY contributed to conception and design of the study. QX and SH collected data from database. QX and SH performed the statistical analyses. QX and LY wrote the first draft of the manuscript. All authors contributed to the article and approved the submitted version.This work was supported by Chinese National Major Project for New Drug Innovation (2019ZX09201002003), National Natural Science Foundation of China , Shenzhen Science and Technology Foundation  and Shenzhen Key Laboratory Foundation (ZDSYS20200811143757022).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.


Waterlogging was one of the main abiotic stresses affecting maize yield and growth in the North China Plain, while ridge tillage effectually improved soil environment, enhanced crop stress resistance to waterlogging, and increased grain yield of waterlogged maize. In order to explore the responses of nitrogen (N) efficiency and antioxidant system of summer maize to waterlogging stress under different tillage, a field experiment was conducted to explore N use efficiency, leaf activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), and malondialdehyde (MDA) content of waterlogged maize Denghai 605 (DH605) and Zhengdan 958 (ZD958) under different tillage system (ridge planting and flat planting). Our results showed that ridge tillage was beneficial to ameliorate waterlogging damages on antioxidant system by increasing SOD, POD, and CAT activities, and decreasing MDA content. Moreover, ridge tillage significantly increased N efficiency of waterlogged maize. N translocation amount (NTA), N translocation efficiency (NTE), N contribution proportion (NCP), N harvest index (NHI), and N use efficiency (NUE) of waterlogging treatment under ridge planting system (W-V3+R) for DH605 was increased by 108%, 69%, 60%, 8% and 16%, while ZD958 increased by 248%, 132%, 146%, 13% and 16%, respectively, compared to those of waterlogging treatment under flat planting system (W-V3). Ultimately, ridge tillage led to a significant yield improvement by 39% and 50% for DH605 and ZD958, respectively, compared to that of W-V3. In conclusion, ridge tillage was conducive to retard leaf aging, and enhance nitrogen efficiency, thereby resulting in a yield improvement of waterlogged summer maize. With global climate change, the frequency and intensity of extreme precipitation events is increased . WaterloWaterlogging mainly happened in areas with high groundwater level, poor drainage and ventilation . Under fVisibly, ridge tillage can effectually improve N efficiency and antioxidant system, and enhance crop resistance to abiotic stresses. However, very little attention has been given to assess the physiological mechanism of ridge planting to regulate yield and growth of waterlogged maize from the aspects of N efficiency and antioxidant system. In this study, we performed a field experiment to explore waterlogging stresses on antioxidant enzymes activities, MDA content, N accumulation and distribution, and N use efficiency of summer maize under different planting system, which will illuminate the regulation mechanism of ridge tillage to antioxidative system and N efficiency of waterlogged summer maize.−1 organic matter, 0.9 g kg−1 total N, 50.7 mg kg−1 rapidly available P, and 86.2 mg kg−1 rapidly available K before the experiment. The effective accumulated temperature was 1,740.9 °C d and 1,710.5 °C d, and the mean total precipitation was 356.0 and 378.5 mm during the growth periods of summer maize in 2014 and 2015, respectively. Summer maize hybrids Denghai605 (DH605) and Zhengdan958 (ZD958), the most widely planted hybrids in China, were used as experimental materials. Maize was sown on June 16 at 67,500 plants ha−1 in both experimental years. Experimental treatments were as follows: tillage treatments , and water treatments . Details of experimental treatments  maintained by the State Key Laboratory of Crop Biology of Shandong Agricultural University, China in 2014 and 2015. This area belongs to a temperate continental monsoon climate. The experimental topsoil (0–20 cm) type is brown loam with 10.7 g kgeatments , accordieatments . There wAt the sixth leaf stage (V6), the twelfth leaf stage (V12), the tasseling stage (VT), the milk stage (R3), and the physiological maturity stage (R6), according to –1) = grain yield/total N uptake amount by plantNUE (kg kgNHI (%) = grain N amount/total N amount of plantNTA (g/plant) = total N amount at VT – total N amount excluding grain N amount at R6NTE (%) = NTA/ total N amount at VTNCP (%) = grain N amount/NTAAt V6, V12, VT, R3 and R6, the functional leaves from three plant samples were obtained to analyze the activities of Superoxide dismutase (SOD), Peroxidase (POD), and Catalase (CAT), and Malondialdehyde (MDA) content, according to the method we described previously .At R6, 30 ears were harvested from three rows at the center of each plot to analysis yield, and yield components. Each treatment was repeated three timesAnalysis of variance (ANOVA) was performed according to the general linear model procedure of SPSS . The least significant difference (LSD) between the means was estimated at the 95% confidence level.The activities of leaf antioxidative enzyme were significantly changed after waterlogging. The activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) for waterlogging treatments were lower than those in the CK treatment at the corresponding growth stages. The overall trend of SOD, POD and CAT activities was consistent among the treatments. Overall, the activities increased firstly and then decreased at growth period. The activities of SOD, POD and CAT of W-V3 (waterlogging treatment under flat planting) for DH605 were averagely significantly decreased by 30%, 25% and 37% for each developmental stage, respectively, compared to those of CK. Those of W-V3 for ZD958 were averagely decreased by 33%, 29% and 37% respectively, compared to those of CK. However, ridge tillage effectually mitigated waterlogging stress on antioxidative enzyme activities. The activities of SOD, POD, and CAT for DH605 under W-V3+R (waterlogging treatment under ridge tillage) were averagely significantly increased by 15%, 24% and 29% for each developmental stage, respectively, compared to those under W-V3. Those of ZD958 under W-V3+R were averagely increased by 22%, 28% and 39% respectively, compared to those under W-V3 .In addition, the malondialdehyde (MDA) content was significantly increased after waterlogging at the corresponding growth stages, compared to CK. However, ridge tillage was beneficial to alleviate the increase of MDA content induced by waterlogging. The MDA content of W-V3+R for DH605 and ZD958 was averagely decreased by 17% and 15% for each developmental stage, respectively, compared to that of W-V3 .The N accumulation of DH605 and ZD958 were averagely reduced after waterlogging by 23% and 33% for each developmental stage across years, respectively, compared to that of CK. Following waterlogging stress, there was an improved N accumulation of waterlogged summer maize under the ridge tillage at each stage. N accumulation of W-V3+R for DH605 and ZD958 was improved by 18% and 29% on the average, respectively, compared to that of W-V3 . FurtherThe N efficiency of summer maize was obviously declined due to waterlogging. N translocation amount (NTA), N translocation efficiency (NTE), N contribution proportion (NCP), N harvest index (NHI), and N use efficiency (NUE) of W-V3 for DH605 were 76%, 64%, 63%, 15% and 20% lower, while those of ZD958 were reduced by 79%, 62%, 64%, 14% and 14% across years, respectively, compared to those of CK. However, ridge tillage helped to improve N efficiency of waterlogged maize. NTA, NTE, NCP, NHI, and NUE of W-V3+R for DH605 were 108%, 69%, 60%, 8% and 16% higher than those of W-V3, respectively. Those of W-V3+R for ZD958 were enhanced by 248%, 132%, 146%, 13% and 16% across years, respectively, compared to those of W-V3 . The redWaterlogging led to soil oxygen deficiency, resulting in the inhibition of nutrient uptake and transportation, and the decrease of grain yield . NitrogeThe inhibition of N uptake and transportation would result in the breakdown of soluble protein and the increase of MDA content, leading to the disorder of reactive oxygen species (ROS) scavenging system, and the acceleration of leaf senescence . ROS scaThe results from the present study showed that ridage tillage was a viable approach to improve N efficiency and leaf antioxidative system of waterlogged maize. The potental areas of future study should focus on the regulation mechanism of ridge farming analyzed from the aspects of root growth, and soil chemicophysical properties, etc.Ridge tillage was conducive to improve antioxidant enzymes activities and nitrogen accumulation and distribution, and decrease MDA content, and thus enhancing nitrogen efficiency, resulted in yield increase of waterlogged summer maize.10.7717/peerj.11834/supp-1Supplemental Information 1Click here for additional data file.


Cardiovascular disease (CVD) has long been the leading cause of death worldwide, and myocardial infarction (MI) accounts for the greatest proportion of CVD. Recent research has revealed that inflammation plays a major role in the pathogenesis of CVD and other manifestations of atherosclerosis. Overwhelming evidence supports the view that macrophages, as the basic cell component of the innate immune system, play a pivotal role in atherosclerosis initiation and progression. Limited but indispensable resident macrophages have been detected in the healthy heart; however, the number of cardiac macrophages significantly increases during cardiac injury. In the early period of initial cardiac damage , numerous classically activated macrophages (M1) originating from the bone marrow and spleen are rapidly recruited to damaged sites, where they are responsible for cardiac remodeling. After the inflammatory stage, the macrophages shift toward an alternatively activated phenotype (M2) that promotes cardiac repair. In addition, extensive studies have shown the therapeutic potential of macrophages as targets, especially for emerging nanoparticle-mediated drug delivery systems. In the present review, we focused on the role of macrophages in the development and progression of MI, factors regulating macrophage activation and function, and the therapeutic potential of macrophages in MI. Despite advances in prevention, diagnosis, and treatment, cardiovascular disease (CVD) remains the primary cause of death worldwide, and myocardial infarction (MI) makes the greatest contribution to CVD . Furtherα), interferon-gamma (IFN-γ), and lipopolysaccharide (LPS) are the leading subtypes that initially respond to the removal of dead cells and cellular and extracellular matrix (ECM) debris [Previous studies have demonstrated that vertebrate zebrafish is capable of complete cardiac regeneration –7, while) debris , 17. In ) debris , 19. TheIn the present review, we discuss recent findings on the association of macrophages with the development of post-MI. More specifically, we focus on the phenotypes and functions of macrophages in a steady state and during MI, as well as possible mechanisms underlying macrophage polarization in the heart. Importantly, we discuss potential therapeutic strategies to improve injury control and functional recovery by modulating macrophage polarization, which involves self-assembly/engineered extracellular vesicles (EVs), nanomedicine, and stem cells Fig. .Fig. 1Mβ (IL-1β), and IL-3 [l° CCR2− CX3CR1hi patrolling monocytes (CD14l°CD16+ in humans), which are responsible for surveying the vascular lumen and for clearing dead cells and cellular and ECM debris, and (2) Ly6Chi CCR2+ CX3CR1l° inflammatory monocytes (CD14hiCD16− in humans), which produce proinflammatory cytokines, such as IL-1, IL-10, and TNF-α. Over the past 40 years, it has been accepted that all forms of macrophages, including resident macrophages, originate from monocytes; however, this has been challenged in the past few years [l°CCR2− and MHC-IIhiCCR2− cells. The others are known as circulating monocyte-derived MHC-IIhiCCR2+ cardiac resident macrophages that are responsible for inflammation initiation  and also regulate organ development and function . Macrophand IL-3 , 23, andand IL-3 : 1) Ly6ew years . Recent ew years , 26, 27.ew years –32. In rew years , 29. It ew years , 31, 33, Ly6ew yein vitro) or microenvironmental milieu (in vivo). Macrophages not only possess the essential functions of phagocytic killing of pathogens and antigen presentation activating an adaptive immune response but also maintain tissue homeostasis by eliminating dead cells, cellular debris, and ECM debris and by promoting adaptive remodeling of the ECM [in vitro stimulation, surface molecule expression pattern, secretory profile, and function. Although this distinction has deficiencies in adequately including the entire macrophage biological complexity, it provides a common scheme to classify macrophage function. In addition, the switch of macrophage polarization is tightly regulated by signaling networks at the transcriptional and translational levels ; TNF-α; chemokine (C-C motif) ligand 2 (CCL2), CCL15, and CCL20; C-X-C motif chemokine 9 (CXCL9), CXCL10, and CXCL11; and CD80 and CD86 [β, ultimately leading to the activation of NF-κB to induce M1 polarization [γ in an autocrine manner to activate these macrophages [via the JAK/STAT signaling pathway, favoring M1 polarization [γ, which is produced by natural killer (NK) cells responding to stress and infections, can promote macrophages to produce proinflammatory cytokineswhich in turn increase the killing ability of NK cells [γ expression are more susceptible to protozoal and certain kinds of bacterial or viral infections [via an exogenous TLR ligand can clear the parasite completely [M1 macrophages are characterized by increased microbicidal or tumoricidal capacity and secrete high amounts of proinflammatory cytokines and mediators. Classically activated M1 macrophages are triggered by IFN-and CD86 –39. M1 mrization . In addirophages . IFNs merization , 42. IFNNK cells . Typicalfections . Furthermpletely . Finallyβ, CD163, CD206, chil3 chitinase-like 3 (known as Ym-1), resistin-like-α (known as Fizz1), arginase 1 (Arg-1), and IL-10 to promote cell proliferation, collagen formation, and tissue repair [via STAT6 and STAT3 [γ, Krüppel-like factor 4 (KLF-4), and HIF-2α also mediate the induction of the M2 phenotype [γ receptors and TLR stimulation can trigger M2b macrophages, and GC, IL-10, or TGF-β ligands are responsible for M2c activation [M2 macrophages are characterized as having wound healing and proliferative properties. M2 macrophages, which are activated by the stimulation of IL-4, IL-13, or IL-21, are involved in wound repair, homeostasis, and tumor metastasis and tumor promotion. They also secrete anti-inflammatory cytokines –47. M2 mnd STAT3 –50. IRF4henotype . M2 macrtivation . In functivation .l°CCR2− and MHC-IIhiCCR2− cells, macrophages derived from fetal monocytes and macrophages derived from adult monocytes [Cardiac resident macrophages, which account for 6% to 8% of the noncardiomyocyte population according to the data from healthy adult mouse heart, are indispensable for maintaining the cardiac homeostasis and neonatal heart regeneration . The lononocytes , 54, 55.− macrophages play a major role in the coronary development and cardiac repair. Deletion of CCR2− macrophages in the embryonic period could cause abnormal remodeling, diminished LV systolic function, larger LV chamber dimensions, and increased akinetic myocardium [+ cardiac resident macrophages are abundant in proinflammatory genes involved in inflammation initiation and exhibit an M1 polarization profile to maintain cardiac homeostasis. Interestingly, more substantial contributions of circulating monocytes have been observed in the aging heart, indicating that circulating blood monocytes may differentiate into CCR2− macrophages [− macrophages with age, as demonstrated by Molawi et al. [As noted, yolk sac–derived CCR2ocardium . Circularophages , 58, whiα, IL-1, and IL-6) and exacerbate local inflammation [Atherosclerosis is considered to be a chronic inflammatory disease. Perturbation of lipid metabolism and local inflammation are the two major causes in the pathogenesis of atherosclerosis, including cells such as platelets, which play an initiating role in the development of atherosclerosis and the atherosclerotic plaque , monocytammation . In addiammation . Next, mammation . The apoammation .MI is defined as pathologically myocardial cell death resulting from prolonged myocardial ischemia. MI is the leading cause of mortality and disability worldwide  and resuhi monocytes and macrophages [hi monocytes are recruited to the infarcted site via MCP-1. Its receptor, CCR2 [α, IL-1β, and proteases, originating from activated Ly-6Chi/M1, are released in the damaged site, which contributes to the clearance of dead cells and debris in the infarcted region through the activated M1 macrophages. The process of phagocytosis is indispensable for proper initiation of the wound repair after MI [MI can lead to necrosis of cardiac myocytes within a few minutes. To maintain tissue integrity and function, inflammatory cells, including neutrophils and macrophages, are activated. After MI, neutrophils are the first immune cells to occur in the infarcted area in large numbers and are responsible for clearing cellular debris and further recruiting leucocytes such as Ly-6Crophages , 68. Monrophages . The twoor, CCR2 , and corafter MI . HoweverMI results in necrosis of cardiac myocytes within a few minutes, contributing to chamber dilatation, contractile dysfunction, and eventually heart failure. The regenerative capacity of mammals persists for only a short period , 9. Therhi monocytes and M1 macrophages are replaced by the Ly-6Clow monocytes and M2 macrophages. The proliferative phase after MI origin from the macrophages shifts from inflammatory (M1) to reparative phenotypes (M2). M2 macrophages gradually dominate in the infarcted sites. Then, M2 macrophages establish an anti-inflammatory environment by downregulating inflammatory cytokines and upregulating anti-inflammatory cytokines, such as IL-10, VEGF, and TGF-β [β and IL-10 can trigger myofibroblasts to produce collagen, and VEGF promotes cell proliferation and blood vessel development. The lack of the Trib1 gene in the myocardial-infarcted mouse model presented selective deletion of M2 macrophages, which resulted from an impaired ability to form M2 macrophages in the spleen, liver, and adipose tissue. Compared with a control group, diminished reparative function after MI, similar to frequent cardiac rupture, was observed. In addition, IL-4 has been demonstrated to improve the post-MI prognosis of wild-type mice with an increased number of M2 macrophages. Together, M2 macrophages play a vital role in facilitating myocardial wound healing in adult murine heart [LysMCre mice, Galuppo et al. reported that the glucocorticoid receptor in macrophages critically determines post-MI repair by regulating myofibroblast differentiation in the infarct microenvironment during the early phase of wound healing [From days 4 to 14 post-MI, Ly-6Cnd TGF-β . TGF-β ane heart . Moreove healing .As noted, the most timely and effective treatment involves amelioration of myocardial ischemia and restriction of the size of MI. Ischemia/reperfusion (I/R) such as percutaneous coronary intervention and intravenous thrombolysis has so far been the principal or only strategy for MI treatment, thereby promptly restoring blood supply . Howeverγ production [Though M1 macrophages are believed to damage the heart in the early period of reperfusion by releasing reactive oxygen species, inflammatory mediators, and proteases , 80, somoduction . M1 macrIn vivo experiments have shown that transplantation of M2b macrophages into the myocardium that had been subjected to I/R injury improved cardiac function and reduced the cardiac fibrosis and myocardial remodeling caused by I/R injury [M2 macrophages, polarized by Th2 cytokines and characterized by the production of high levels of anti-inflammatory cytokines and pro-fibrogenic factors, exhibit anti-inflammatory and tissue repair properties. Accumulated evidence has demonstrated that M2 macrophages play an important role in alleviating myocardial I/R injury. A previous study reported that alternatively activated M2 macrophages by Chemerin15 protect against myocardial I/R injury in mice by significantly suppressing proinflammatory cytokines and markedly increasing the level of anti-inflammatory cytokine IL-10 . In receR injury . Yue et R injury .in vitro and in vivo [It has been reported that stem cells and stromal cells could be utilized for the treatment of myocardial injury by modulating macrophage polarization. Cardiosphere-derived cells (CDCs), essentially cardiac stromal cells (CSCs), represent a promising stem cell source for repairing damaged heart tissue –88. Thes in vivo .EVs have been demonstrated to play a crucial role in cell-cell communication during different pathological and physiological processes , 97. EVsMSCs have long become a promising therapeutic strategy for ischemic heart disease, although the mechanism remains elusive. Several recent studies have implicated that MSC-exo could polarize macrophage to create an anti-inflammatory environment under myocardial I/R injury or MI , 103. MSvia EV transfer of miR-26a to modulate the expression of MerTK and C1qa [in vitro and confer cardioprotection in vivo by minimizing infarct size, decreasing the total number of CD68+ macrophages, and inducing macrophages to develop into a distinct subtype [CDC or CDC-derived EVs polarize the macrophage into a special phenotype that is highly phagocytic and anti-inflammatory to display cardioprotection , 106. Deand C1qa . Moreove subtype . Another subtype . Further subtype .via transferring miRNAs, especially the miR-146a, which downregulates target genes IGF2BP1 and HuR in recipient cells, thereby promoting the progression of atherosclerosis [in vivo experiments revealed a lower incidence of cardiac rupture and improved cardiac function in the miR-155–deficient mice compared with controls [Nguyen et al. reported that atherogenic macrophage-secreted EVs can inhibit macrophage migration clerosis . Moreovecontrols . In addicontrols . Furthercontrols .Although EVs have been demonstrated to be an attractive therapeutic approach to ischemic heart disease, low retention and short-lived therapeutic effects remain significant challenges. To reduce off-target delivery, engineered exosomes and prior blocking of endocytosis of exosomes by macrophages have been utilized to enhance the delivery efficiency of exosomes to specific cells, offering therapeutic benefit –116. ThuTo improve the safety and efficiency of therapeutic agents, specially designed nanocarriers, including liposomes, polymeric nanoparticles, and complexes, have been widely applied, particularly in the cardiovascular and oncology fields. These nanocarriers containing active substances, such as siRNAs or statins, effectively polarize macrophages into the anti-inflammatory phenotype or mediate gene expression in macrophages Table . Increas+ macrophages and the level of anti-inflammatory cytokines, such as TGF-β and IL-10, and downregulates the expression of proinflammatory markers at the same time, which promotes angiogenesis and halts ventricular dilatation and adverse remodeling in a rat model of acute MI [Liposomes are microscopic phospholipid bubbles, which include one or more biocompatible lipid bilayers with an aqueous core inside, ranging from a few hundreds to thousands of nanometers in size –123. Lipacute MI . Moreoveacute MI .Polymeric nanoparticles (NPs) mainly consist of biodegradable and biocompatible polymers such as natural polymers , polylactide, and poly (PLGA) and are heterogeneous in size, often ranging from a few tens to thousands of nanometers in diameter . BecauseMonocytes/macrophages have been known as targets that are modulated by polymeric NP– and complex-based carriers in the field of cardiac repair and have caught special attention in the past few years. Recent studies have demonstrated that systemic administration of siRNA against messenger (mRNA) loaded into NPs imparts beneficial outcomes in cardiac repair , 116. Acγ (PPARγ) agonist, shows a marked impact on monocyte and macrophage polarization, transferring them into the anti-inflammatory subtype [−/− mouse model showed a significant reduction in the number of fibrous caps by decreasing proinflammatory monocytes along with a moderate increase in anti-inflammatory phenotypes [Furthermore, NPs containing pioglitazone have been demonstrated to modulate monocyte/macrophage subtype , 132. Pi subtype . Matoba  subtype . In a siMoreover, the timely endocytosis of dead cells by macrophages can trigger the anti-inflammatory response and transform M1 macrophages into M2 macrophages. Given these findings, a theranostic nanosystem with mimicking apoptosis was established and showed a remarkable ability in resolving inflammation and promoting cardiac repair .Macrophages play an indispensable role in the mammalian heart and respond to both the post-MI regeneration and repair by mediating inflammation and immunity. M1 and M2 macrophages are indispensable in repairing the myocardium and in retaining functional architecture. Different populations of macrophages respond to a special stage after MI. Several studies have focused on the mechanism of macrophage polarization that is related to the cardioprotective potential. With advancement in therapeutic approaches, especially the burgeoning NP-mediated drug delivery system, modulating macrophage activation to guide cardiac repair and regeneration is now a promising therapeutic strategy. These strategies are also available to other CVDs associated with macrophages, such as atherosclerosis and myocarditis.As this review highlights, the roles of macrophages in terms of cardiac repair and regeneration are complex. To better understand the mechanisms of macrophages during the various stages of acute and chronic myocardial disease, further research is warranted. Moreover, although specially designed nanocarriers have been applied to improve the efficiency of therapeutic agents and a major breakthrough has been reached, many challenges remain, including the off-target delivery and security issues, which require additional investigation.


SOD1 is the major superoxide dismutase responsible for catalyzing dismutation of superoxide to hydrogen peroxide and molecular oxygen. It is well known as an essential antioxidant enzyme for maintaining cellular redox homeostasis. SOD1 dysregulation has been associated with many diseases, including amyotrophic lateral sclerosis (ALS), cancer, accelerated aging, and age-related diseases. Recent studies also revealed that SOD1 can serve as a regulatory protein in cell signaling, transcription, and ribosome biogenesis. Notably, SOD1 is localized in the nucleus under both normal and pathological conditions, contributing to oxidative stress response and growth control. Moreover, increasing evidence points to the importance of nuclear SOD1 in the pathogenesis of ALS and cancer. As much as one to two percent of oxygen consumed by mammalian cells is estimated to be converted to superoxide [2O2), which is comparatively less reactive. Both superoxide and hydrogen peroxide are key reactive oxygen species (ROS) that play important roles in regulatory processes and stress responses [Superoxide , hydroxyl radical (HO•), and reactive aldehydes, can oxidize DNA, RNA, lipids, and proteins, producing not only aberrant regulatory responses to alter cellular processes, but also reversible and irreversible damage to cells and tissues . Oxidati2•− + 2H+ → H2O2 + O2 [2+/Zn2+ enzyme; SOD2, a Mn2+ enzyme; and SOD3, a distinct Cu2+/Zn2+ enzyme [Because of its severely deleterious effects, superoxide is rapidly dismutated through enzyme-catalyzed reactions into hydrogen peroxide, that in turn is converted to water and molecular oxygen. Superoxide dismutases (SODs) are enzymes responsible for scavenging superoxide into hydrogen peroxide and molecular oxygen via the following chemical reaction: 2O2O2 + O2 . All cel+ enzyme . The thr+ enzyme , whereas+ enzyme . SOD1 caSod1 knockout in mice fails to recapitulate motor neuron disease phenotypes, fALS mutations in SOD1 are thought to give rise to a gain-of-function causing motor neuron degeneration [SOD1 was discovered more than 50 years ago by biochemical purification of the enzymatic activity catalyzing dismutation of superoxide ions . SOD1 haneration ,14.Sod1 knockout attenuates these oncogene-driven proliferations, but not normal proliferation of the mammary gland associated with pregnancy or other normal proliferative tissues, such as skin and gastro-intestinal tract epithelium [Sod1 knockout significantly reduced lung tumor burdens due to inhibition of cancer cell proliferation [Sod1 knockout does not have discernible effect on proliferation of normal lung cells or other tissues [SOD1 is overexpressed in many human cancers, including non-small-cell lung cancer (NSCLC), breast cancer, and nasopharyngeal carcinoma (NPC) ,16,17,18feration . In cont tissues . These oGiven its role in cancer cell proliferation, SOD1 represents a promising anticancer drug target. Several distinct small molecule inhibitors of SOD1 have been identified that are under preclinical and clinical evaluations, including ATN-224 and LCS-1. ATN-224, a copper chelator, is the most advanced SOD1-targeting agent that has been tested in Phase 1 and Phase 2 clinical trials for patients with relapsed prostate cancer ,24. RegrSod1 knockout mice exhibit widespread oxidative stress, elevated oxidative DNA damage, and shortened lifespans [Sod1 knockout increased tumor incidence [ifespans . These mifespans , likely ncidence . These oncidence . In the ncidence .https://www.proteinatlas.org/ENSG00000142168-SOD1/tissue, accessed on 18 January 2022). Nuclear SOD1 is particularly enriched in the breast, cerebellum, the dermis layer of the skin, and testis.Early studies using subcellular fractionation showed that SOD1 is present in the cytosol, nucleus, peroxisomes, and mitochondrial intermembrane space of rat liver . Immunof2O2 levels [2O2 [2O2 was also shown to elicit SOD1 nuclear accumulation in human fibroblasts and HeLa cells [ATM-deficient fibroblasts [In yeast and human fibroblasts, SOD1 becomes more enriched in the nucleus in response to oxidative stress elicited by increased H2 levels . In yeas2 levels . ATM kinels [2O2 . In yeasels [2O2 . In humaLa cells ,32. Thisroblasts , suggestroblasts . It shouProtein transport into the nucleus is mediated by short amino acid sequences known as nuclear localization sequences (NLSs), which are recognized by importins . SimilarIn yeast cells, hydrogen peroxide stimulates SOD1 entry into the nucleus and regulates the expression of oxidative-stress-responsive genes. SOD1 target genes include those involved in oxidative stress resistance , DNA damage repair and genomic stability maintenance , DNA replication stress , and iron/copper homeostasis . Consistent with a role of nuclear SOD1 in DNA damage repair and maintenance of genomic stability, nuclear SOD1, not cytoplasmic SOD1, was shown to provide protection against DNA damage in yeast and chicken cells ,34.Mechanistically, yeast SOD1 was shown to bind to promoters of its target genes, indicating that SOD1 acts as a transcription factor . In a lahttps://www.proteinatlas.org/ENSG00000142168-SOD1/pathology, accessed on 18 January 2022). Consistently, SOD1 is found enriched in the nuclei of breast and lung cancer cells [Sod1 knockout mouse NSCLC cells [Sod1 knockout mouse NSCLC cells, including NAC (N-acetyl-l-cysteine), MnTBAP and Tempol (superoxide dismutase mimetics), GSH-MEE (a membrane permeable GSH derivative), and Trolox  [SOD1 is overexpressed in many human cancers, including non-small-cell lung cancer (NSCLC), nasopharyngeal carcinoma (NPC), and breast cancer ,16,17,18Sod1 knockout blunts nucleolar hypertrophy in Kras/TP53-driven NSCLC tumors in mice [Ribosome biogenesis occurs primarily in the nucleolus . It invo in mice . These oSOD1 as a transcription factor is also thought to contribute to cancer development and maintenance. Due to aberrant metabolism, cancer cells contain much higher ROS content than that found in normal cells. Although moderate levels of ROS stimulate cancer cell proliferation, excess ROS damages macromolecules, such as DNA and lipids. As described above, SOD1 promotes expression of many genes involved in oxidative stress response. Ribonucleotide reductase (RNR), for example, is a SOD1 transcriptional target ,32. ThisALS is characterized by accumulation of SOD1 protein aggregates that are thought to be toxic to motor neurons ,47. DespIn PBMCs from sALS patients, SOD1 aggregates prevent hydrogen peroxide-induced ATM/CHK2-dependent translocation of SOD1 into the nucleus, which is correlated with elevated oxidative DNA damage . One posSOD1 has been known to be localized in the nucleus for over two decades. Only recently did its nuclear functions come to light. Nuclear SOD1 engages in non-canonical biochemical functions, such as regulation of transcription and ribosome biogenesis, both processes important for oxidative defense and growth. Importantly, nuclear SOD1 functions provide new insights into pathological mechanisms of SOD1 in cancer and ALS. Moving forward, it will be important to further understand the molecular details of these new functions in more physiologically relevant in vivo models, which will be necessary for improving prevention and treatment of these diseases. Given the difficulty to treat KRAS-driven NSCLC, targeting nuclear SOD1 should open a new avenue for discovery and development of new targeted therapeutic agents for management of this lethal malignancy.


Herbal medicine is one of crucial symbols of Chinese national medicine. Investigation on molecular responses of different herbal strategies against viral myocarditis is immeasurably conducive to targeting drug development in the current international absence of miracle treatment.  Literature retrieval platforms were applied in the collection of existing empirical evidences for viral myocarditis-related single-herbal strategies. SwissTargetPrediction, Metascape, and Discovery Studio coordinating with multidatabases investigated underlying target genes, interactive proteins, and docking molecules in turn. Hedysarum Multijugum Maxim), Yuganzi (Phyllanthi Fructus), Kushen (Sophorae Flavescentis Radix), Jianghuang (Curcumaelongae Rhizoma), Chaihu (Radix Bupleuri), and Jixueteng (Spatholobus Suberectus Dunn) meet the requirement. There were 11 overlapped and 73 unique natural components detected in these herbs. SLC6A2, SLC6A4, NOS2, PPARA, PPARG, ACHE, CYP2C19, CYP51A1, and CHRM2 were equally targeted by six herbs and identified as viral myocarditis-associated symbols. MCODE algorithm exposed the hub role of SRC and EGFR in strategies without Jianghuang. Subsequently, we learned intermolecular interactions of herbal components and their targeting heart-tissue-specific CHRM2, FABP3, TNNC1, TNNI3, TNNT2, and SCN5A and cardiac-myocytes-specific IL6, MMP1, and PLAT coupled with viral myocarditis. Ten interactive characteristics such as π-alkyl and van der Waals were modeled in which ARG111, LYS253, ILE114, and VAL11 on cardiac troponin (TNNC1-TNNI3-TNNT2) and ARG208, ASN106, and ALA258 on MMP1 fulfilled potential communicating anchor with ellagic acid, 5α, 9α-dihydroxymatrine, and leachianone g via hydrogen bond and hydrophobic interaction, respectively.  Six single-herbal medicines consisting of Huangqi ( The comprehensive outcomes uncover differences and linkages between six herbs against viral myocarditis through component and target analysis, fostering development of drugs. Herbal medicine is the keystone to uphold the existence and development of Traditional Chinese Medicine (TCM); besides China, herbs are also widely applied to improve human health in Sumer and ancient Egypt for thousands of years –4. CurreGalega officinalis, is a prevalent first-line treatment for diabetes [Cymbopogon citratus) and dampens polyamine biosynthesis and cell growth [Artemisia capillaris is a famous traditional Chinese herb, and its extract enynes are responsible for the effect of anti-hepatitis B virus significantly inhibiting viral DNA replication [μg/mL doses of Sambucus nigra fruit extract, the titer and protein synthesis of H9N2 influenza virus are palpably decreased in the human epithelium cell which reflects the herb interferes with either entry of viruses or release of the virus particle [There are several clinical and experimental evidence of herbal medicinal efficacy on angiocardiopathy, diabetes, cancer, and other inflammatory or viral diseases. A cardiovascular investigation involving 781 patients indicated that the intake of standardised garlic extract (600 to 900 mg per day) is coupled with 0.41 mmol/L reduction in serum cholesterol level [diabetes . A priordiabetes . Epidemidiabetes . A rat sdiabetes . Implical growth . The stul growth , 15. Artlication . Throughparticle .Myocarditis is an inflammatory cardiomyopathy, symptoms of which include irregular heartbeat, pectoralgia, shortness of breath, and impaired ability to exercise . CompareAlthough abundant achievements clarify herbs' effectiveness on viruses, the differences of single-herbal strategies have been seldom pursued, especially against VMC. Herein, relying on open-resource platforms and bioinformatics methods, we designed and executed an investigation to compare the chemical compositions, molecular targets, and their interactions of distinct single-herbal strategies potentially coupled with treatment of VMC and attempted to provide inspirations against VMC.https://pubmed.ncbi.nlm.nih.gov) and Web of Science (http://www.webofscience.com), free retrieval engines about the biomedical literature [To comprehend medical strategies of single herb that treat with a single herb and have been revealed for antiviral activity on VMC, information search was performed by PubMed  displays twelve essential properties such as herbal distribution, absorption, excretion, and metabolism and is invoked to completely view herbal medicines based on the framework of systems pharmacology [https://pubchem.ncbi.nlm.nih.gov) as a public repository presents mostly small and also larger molecule data such as chemical structures, safety, and toxicity and is ordinarily applied to chemical biology investigation and drug discovery [The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP)  is an analysis platform of ligand-based target prediction on a bioactive small molecule and delivers services to more than one hundred countries worldwide [SwissTargetPrediction (orldwide . Taking orldwide . The herhttps://www.genecards.org) as an integrative and searchable database supplies inclusive, authoritative compendium of annotative information about human genes. The knowledge database integrates gene-related data from nearly one hundred and fifty web sources, embodying genomic, transcriptomic, proteomic, genetic, clinical, and functional information [GeneCards  provides a comprehensive annotation and analysis of gene list to experimental biologists [p value less than 0.05 was defined as statistically significant.The web-based resource Metascape (ologists . The enrhttp://www1.rcsb.org/) archives and shares experimentally determined 3D structures of nucleic acids, proteins, and complex assemblies derived from crystallography, nuclear magnetic resonance spectroscopy, and electron microscopy [Homo sapiens setting checked. The receptor-ligand interaction between the target protein and active component was carried out by the software BIOVIA Discovery Studio v16.1.0 [The Protein Data Bank (PDB) (croscopy . We used v16.1.0 .Hedysarum Multijugum Maxim), Yuganzi , Kushen , Jianghuang , Chaihu , and Jixueteng  as the qualified objects to analyse. In line with the preestablished criteria, we collected 17 , 14 , 36 , 2 (stigmasterol and CLR), 13 , and 19  constituents in HQ, YGZ, KS, JH, CH, and JXT in turn  3514, 3290, 3979, 556, 3328, and 3811 terms and 393, 366, 418, 23, 370, and 416 pathways in all targets of HQ, YGZ, KS, JH, CH, and JXT in turn, as well as 1553, 1691, 1956, 117, 1540, and 2076 terms and 255, 253, 319, 0, 229, and 335 pathways in VMC-related targets of that. By analysing the top 10  located in heart, as significant (p < 0.001) as KS targeting whole AXL, CHRM2, CHRNA5, S1PR3, UTS2R, KCNA5, LNPEP, PDE3A, PLA2G5, SCN5A, TNNC1, TNNI3, TNNT2, TNKS, MAPKAPK2, and TNNI3K and VMC-related CHRM2, CHRNA5, SCN5A, TNNC1, TNNI3, and TNNT2 (p < 0.01) by HQ targeting whole AXL, MMP1, PLAT, RGS4, and PLK2, CH targeting whole AXL, F2R, and MMP1, and JXT targeting whole AXL, IL6, MMP1, PLAT, PLK2 and VMC-related IL6, MMP1, and PLAT , FABP3 , TNNC1-TNNI3-TNNT2 (PDB ID: 1j1e), SCN5A , IL6 , MMP1 , PLAT , and their binding components revealed reactive CHRM2 (PDB ID: 4mqs), FABP3 (PDB ID: 3wxq), TNNC1-TNNI3-TNNT2 (PDB ID: 1j1e), and MMP1 (PDB ID: 3shi) with respective 12, 3, 4, and 9 components . The typI3-TNNT2  and S4.β, and tumor necrosis factor in the CVB3-infected mice [Previous research has reported that 10-mL HQ oral liquid daily significantly decreases sinus tachycardia, frequent premature ventricular contractions, and supraventricular tachycardia and improves myocardial enzymes and cardiac function indexes compared to placebo daily in 68 VMC children . With inted mice . CH protted mice . Aqueousted mice . Hence, SLC6A2 and responsible for presynaptic noradrenaline reuptake. Between the vasculature, heart, and kidney, it plays an essential role in the distribution of sympathetic activity. Genetic SLC6A2 dysfunction is capable of triggering the postural tachycardia syndrome while the impaired function of cardiac SLC6A2 is familiar in a variety of organic heart disease such as ischemic heart disease, congestive heart failure, and stress-induced cardiomyopathy [NOS2 encodes inducible nitric oxide synthase. Myocardial infiltrating macrophages express high levels of inducible nitric oxide synthase in CVB3-infected male mice [NOS2 contribute to lower viral stocks [NOS2 results in a sudden rise in the mortality of mice with Coxsackievirus infection [NOS2 expression exaggerates myocardial damage [SLC6A4 is active in heart valve development, and its deficiency is conjoined with apparent perivascular, interstitial, and valvular fibrosis [PPARA encodes peroxisome-proliferator-activated receptor alpha whose activation improves experimental autoimmune myocarditis through restraining Th17 cell differentiation under expression inhibition of retinoic acid receptor-related orphan nuclear receptor gamma and phosphorylated signal transducer and activator of transcription 3 in vivo [PPARG encodes peroxisome-proliferator-activated receptor gamma. A small heterodimer partner expressed in the heart can attenuate the hypertrophic response, while changes in inflammation and metabolism are correlated with marked alterations in the mRNA levels of PPARA and PPARG in small heterodimer partner overexpressing cells [CYP2C19 and frequent expression of the corresponding protein cytochrome P450 2C19 have been considered as a protective compensation reaction in chronic Keshan disease, an endemic cardiomyopathy [CYP51A1 encodes lanosterol 14-alpha demethylase. The CYP51A1 deficiency in mice shows heart failure and lethality owing to heart hypoplasia, vasculogenesis, ventricle septum, and epicardial defects [ACHE is involved in regulating levels of acetylcholine which is an anti-inflammatory molecule connected to inflammatory response [CHRM2 encodes muscarinic acetylcholine receptor M2 such that the missense mutation (C722 G) identified in the CHRM2 triggers heart failure, arrhythmia, and sudden death in the patients with dilated cardiomyopathy [Using the TCMSP, PubChem, and SwissTargetPrediction, we screened out 79 components and their 786 potential targets by duplication removing from six single-herbal strategies. The whole 786 targets ranged over 150 VMC-associated symbols. Our priority was to focus on analysing nine common VMC-associated targets including SLC6A2, NOS2, SLC6A4, PPARA, ACHE, CYP2C19, PPARG, CYP51A1, and CHRM2 in six herbal strategies. Sodium-dependent noradrenaline transporter targeted by 9 herbal components is encoded by myopathy . NOS2 enale mice . The higl stocks . Lack ofnfection . But notl damage . Sodium-fibrosis . Peroxisfibrosis . PPARA e in vivo . PPARG eng cells . There ing cells . The perng cells . Besidesng cells . In termng cells , 71. Themyopathy . CYP51A1 defects . Acetylcresponse . CHRM2 emyopathy . In lighin vitro and in living COS-7 cells and may be defensive in human heart failure [What follows is machine learning of prospective targets that refers to functional enrichment, protein interaction, and specific location analyses comparing VMC-associated targets to whole targets in different herbal strategies. In terms of numbers of the abovementioned elements enriched by VMC-associated targets, more than 3000 GO terms and 300 KEGG pathways were recruited by the whole targets of herbal strategies without JH. Our findings demonstrated that cellular response to the nitrogen compound (GO:1901699) and positive regulation of transferase activity (GO:0051347) preferred to be significantly enriched by whole and VMC-associated targets. There is a report that nitric oxide disables the coxsackieviral protease 2A by active-cysteine S-nitrosylation  failure . Histone failure . Respons failure . Moreove failure . Adoptiv failure , while a failure . With vi failure . The P38 failure . The inv failure –86. ThisSRC and EGFR separately encode proto-oncogene tyrosine-protein kinase Src and receptor protein-tyrosine kinase. Under coxsackieviral infection, the viral production in myocytes is reduced by SRC inhibition [CYP51A1, CYP2C19, PPARG, and NOS2 noted earlier, FDFT1 and VDR are responsible for encoding squalene synthase and vitamin D3 receptor, respectively. Ding et al. reported that changes in a network of coexpressed cholesterol metabolism genes encompassing sterol synthesis gene FDFT1 are a characteristic mark of inflammatory stress [Besides single target, multiple targets are the tendency of new pharmaceutical development. We hope that, with the help of the PPI network, examines the role of single target or several targets on the balance of network and its perturbations. In the present work, we discovered that SRC and EGFR as PPI hubs have more than twenty partners possessing interactive potential both in whole and VMC-associated targets of HQ, YGZ, KS, CH, and JXT. hibition . EGFR rehibition . Comparey stress . VDR is y stress . InterfeFABP3) deficiency alleviates myocardial apoptosis and cardiac remodeling, forming a protection from ischemic heart injuries [TNNC1 encodes slow skeletal and cardiac-type troponin C1, and its mutations play an essential role in the development of cardiomyopathy, in which the TNNC1-A8V mutant evokes diastolic disorder through raising the calcium-ion-binding affinity of the thin filament and altering calcium ion homeostasis and cellular remodeling [TNNI3 and SCN5A. Seven of 42 patients with acute myocarditis carry infrequent biallelic nonsynonymous or splice-site variations in cardiomyopathy-related TNNI3 or SCN5A [TNNT2 elevates the cardiomyogenic efficiency of cardiosphere-derived cells to form large cardiomyocytes populations [CHRNA5 encodes neuronal acetylcholine receptor subunit alpha-5. The secretion of proinflammatory cytokine interleukin-1β is significantly decreased by fifty percent in bone-marrow-derived macrophages by comparing CHRNA5 knockout mice with wild-type controls [IL6) whose levels were reduced after receiving antiviral therapy [MMP1) expression and attenuates the myocardial fibrosis and reduces the mortality in mice with chronic myocarditis [PLAT are implicated in strokes and myocardial infarctions and susceptible to bacterial osteomyelitis [The next detail is that specific targets were detected in 31 kinds of tissues and 29 types of cells. Taking significant enrichment as the screening standard, the categories of specific tissues focused by whole targets generally exceed that covered by VMC-associated targets in number. As shown in injuries . TNNC1 emodeling . Cardiacor SCN5A . As a caulations . CHRNA5 controls . In card therapy , 97. Astcarditis . Polymormyelitis . A priormyelitis . This coα, 9α-dihydroxymatrine (MOL006582 index 1), as well as ARG208, ASN106 coupled to conventional hydrogen bond, and ALA258 connected to hydrophobic interaction on MMP1 with leachianone g (MOL006626 index 2). A recent study proved that hydrophobic groups and hydrogen bond acceptors may work in the inhibitory potency of flavonoids existed in herbal products on breast cancer resistance protein [Trypanosoma brucei pteridine reductase 1 or Leishmania major pteridine reductase 1 with chroman-4-one moiety expose their relevance on the compound activity and could be one of the causes of inhibitory effects of chroman-4-one moiety to the two reductases [Shewanella sp. SIB1, to the native or reduced states of insulin is mainly facilitated by hydrophobic interaction [Intermolecular interactions dominate various important physical and chemical properties of herbal components. Correlated with 12, 3, 4, 9 components, and their respective target CHRM2 (PDB ID: 4mqs), FABP3 (PDB ID: 3wxq), TNNC1-TNNI3-TNNT2 (PDB ID: 1j1e), and MMP1 (PDB ID: 3shi), we found that, on human cardiac troponin (TNNC1-TNNI3-TNNT2), amino acid ARG111 showed a conventional hydrogen bond with ellagic acid (MOL001002 index 6), and LYS253, ILE114, and VAL118 individually acted as an interactive anchor of the conventional hydrogen bond and hydrophobic interaction with 5 protein . The intductases . The bineraction . TherefoIn the present work, we collected six single-herbal strategies against VMC and screened out active components and their corresponding targets. Enrichment analysis underlined centric targets fixed in the PPI network and specific targets localized in heart, following annotation of VMC-related symbols. Besides that, a receptor-ligand interaction model clarified the underlying categories of intermolecular interactions and efficient amino acids based on herbal components and targets in the location of heart lesions. These findings may contribute to the development of new treatments and targeted drugs against VMC in the future.


During the COVID-19 pandemic, patients at the John G. Bartlett Specialty practice experienced disruptions in viral load (VL) monitoring due to 1) conversion to telemedicine visits and 2) closure of the onsite lab from March 16-July 13, 2021. We described the impact of the pandemic on VL monitoring. We measured time from all index VLs collected during 3 periods: January 1, 2019 to March 15, 2020 (pre-pandemic); March 16 to July 12, 2020 ; and July 13 to December 31, 2020  until a subsequent VL, 1 year after the index VL, or administrative censoring on December 31, 2020, whichever came first. We classified follow-up time according to these periods (treating period as a time-varying variable). We report hazard ratios (HRs) and 95% Confidence Intervals (CI) from a Cox proportional hazards model comparing the hazard of a VL during the pandemic periods to the pre-pandemic period, stratified by whether the index VL was suppressed (≤200 copies/mL). We tested for interactions between patient characteristics  and period, to investigate differential effects of the pandemic on delayed VL.After 7,760 suppressed VL measurements, median times to subsequent VL during the pre-pandemic, pandemic (closed lab) and pandemic (open lab) periods, were 4.6 (HR=1.0), 8.9 , and 5.8  months respectively. After 1,025 non-suppressed VL measurements, median times to subsequent VL were 2.0 (HR=1.0), 3.9 , and 2.1  months respectively. Time to subsequent VL after an index suppressed VL was less affected by the pandemic for patients who are white; had private insurance; or had no recent cocaine or heroin use. The effect of the pandemic on time to subsequent VL after a non-suppressed index VL did not significantly differ across patient characteristics.Onsite lab closure disrupted VL collection for all groups. Once the onsite lab opened, the pandemic period was still associated with a delay among suppressed patients, but not non-suppressed patients. Further studies are needed to investigate if these delays are associated with lapses in viral suppression. All Authors: No reported disclosures


Manihot esculenta Crantz), 23 tubulin genes (MeTubulins) were identified, which were divided into four main groups based on their type and phylogenetic characteristics. The same grouping generally has the same or similar motif composition and exon–intron structure. Collinear analysis showed that fragment repetition event is the main factor in amplification of cassava tubulin superfamily gene. The expression profiles of MeTubulin genes in various tissue were analyzed, and it was found that MeTubulins were mainly expressed in leaf, petiole, and stem, while FtsZ2-1 was highly expressed in storage root. The qRT-PCR results of the FtsZ2-1 gene under hormone and abiotic stresses showed that indole-3-acetic acid (IAA) and gibberellin A3 (GA3) stresses could significantly increase the expression of the FtsZ2-1 gene, thereby revealing the potential role of FtsZ2-1 in IAA and GA3 stress-induced responses.Filamentous temperature-sensitive protein Z (Tubulin/FtsZ) family is a group of conserved GTP-binding (guanine nucleotide-binding) proteins, which are closely related to plant tissue development and organ formation as the major component of the cytoskeleton. According to the published genome sequence information of cassava ( Microtubules are polymerized by tubulins, which participate in cell division as one of the cytoskeleton systems, while tubulins are encoded by multiple gene families in plants ,2. The tthaliana . Transgeild type .FtsZ gene, most plants contain three functionally complementary genes, FtsZ1-1, FtsZ2-1, and FtsZ2-2, which presumably arose by gene replication of a single ancestral FtsZ gene [FtsZ1-1 or FtsZ2-1 null mutants can cause chloroplast division defects. Compared with the wild type, a decreased number and increased size of chloroplasts in the cell of green leaf were observed. Unlike FtsZ1-1 and FtsZ2-1 mutants, the phenotype of the FtsZ2-2 null mutant is milder, and the number and size of the chloroplast are less variable [FtsZ, homologs of tubulin, were first found in bacteria and involved in the bacterial division as cytoskeletal proteins, which became key proteins of chloroplast division in higher plants with the occurrence of endosymbiotic events ,17,18,19tsZ gene ,23,24,25variable ,27. Durivariable ,29.The cytoskeleton tubulin and FtsZ proteins have similar functions; both are involved in polymer formation and play a key role in cell division, while tubulin and FtsZ proteins have little sequence identity but showed high conformational similarity ,30. Tubuβ-tubulin1 gene under high concentration of auxin stress, while good stability under other abiotic stresses; the expression of α-tubulin2 gene from Hevea brasiliensis was regulated by NaCl, drought, and MeJA [the tubulin gene family during cassava growth, especially the regulation of FtsZ on chloroplast, it is of great significance to research the function of the cassava tubulin gene family to improve the quality of cassava. For this reason, we analyzed the gene characteristics, phylogeny, gene structure, gene repetition, protein motif, and expression profiles in various tissues, of cassava tubulin family members, and the expression of the FtsZ2-1 gene was studied under IAA, MeJA, ABA, GA3, salt, and drought treatments.Cassava is an important cash crop and food source growing in Africa, Asia, and tropical America because of its high starch content in storage root; about 750 million people depend on cassava as food ,36. Meanand MeJA ,48. BaseMeTubulin genes were named from MeTubulin1 to MeTubulin23. Through BLAST  search and HMMER  analysis, 23 genes with tubulin domain were identified and annotated as MeTubulins in the cassava genome. According to the order of screening, 23 Oryza sativa, Hevea brasiliensis, Arabidopsis thaliana, Dioscorea rotundata, and Vitis vinifera, the unrooted phylogenetic trees were constructed by the maximum likelihood (ML) method, and their evolutionary relationship were further analyzed. A total of 115 tubulins contained 31 Hevea brasiliensis, 16 Arabidopsis thaliana, 16 Oryza sativa Japonica, 11 Dioscorea rotundata, and 18 Vitis vinifera, which were divided into groups I, II, III, and IV corresponding to β-tubulin, α-tubulin, γ-tubulin and FtsZ based on the four types of tubulin , one AtTubulin, three VitTubulins, three OsTubulin10, two HbTubulins, and one DrTubulins belonged to group Ib, which generally contains an extra intron in the 5′ untranslated region. Group Ic mainly consisted of dicotyledons, four MeTubulins , two AtTubulins, seven HbTubulins, and three ViTubulins, and only two monocotyledons OsTubulin10, and DrTubulin2. Similarly, group IIa usually contains three introns, including three MeTubulins , together with two AtTubulins, four HbTubulins, two OsTubulins, four VitTubulins, and two DrTubulins. Group IIb generally contains four introns, consisting of three MeTubulins , two AtTubulins, two VitTubulins, two OsTubulins, four HbTubulins, and two DrTubulins. Two MeTubulins (MeTubulin 6 and 16) clustered with two AtTubulins, DrTubulin7, three HbTubulins, OsTubulin4, and ViTubulin13 in group III. Group IV consisted of three MeTubulins , three AtTubulins, three DrTubulins, three OsTubulins, two VitTubulins, and three HbTubulins. HbTubulin4 was excluded from the grouping because it was too divergent. Furthermore, a total of four sister pairs of MeTubulins were showed in the phylogenetic tree, including MeTubulin2–MeTubulin10, MeTubulin8–MeTubulin23, MeTubulin15–MeTubulin21, MeTubulin12–MeTubulin19. Tubulin domain sequence alignment of MeTubulin proteins showed that sequence varied greatly from position 33 to 46 amino acids, but the sequence was identical or similar in the same group  in In order to gain a better understanding of the evolution of the cassava tubulin family, the exon–intron structures of all identified MeTubulins were unevenly distributed on 11 chromosomes, and each chromosome contained 1-3 MeTubulins in MeTubulins by BLASTP and multiple collinearity scan toolkit (MCScanX) methods in MeTubulin genes might be produced by segmental replication events.In total, 23 tubulin family further, five comparative syntenic maps of cassava associated with five representative species were constructed, including two monocots (Oryza sativa and Dioscorea rotundata) and three dicots  (Dioscorea rotundata (7), and Oryza sativa (1) have fewer collinear genes than Vitis vinifera (18), Populus trichocarpa (14), and Arabidopsis thaliana (9). Some MeTubulins have been observed to be associated with more than one syntenic gene pairs, especially cassava and Vitis vinifera, such as MeTubulin9 and MeTubulin17, speculated that these genes might play a significant role in the evolution of the MeTubulin family.To research the evolutionary mechanisms of the cassava inifera) . In geneinifera) . For exaTubulin genes  database under the accession number GSE7951 , which cin genes . We founFtsZ2-1 gene by different abiotic stresses and hormonal treatments , ABA (abscisic acid), GA3, NaCl, and PEG (polyethylene glycol) treatments to research the expression of the eatments . Under 1FtsZ2-1 gene under various stresses was researched. Tubulin genes were screened from the cassava genome, along with six other species, for instance, Arabidopsis thaliana [Dioscorea rotundata [Hevea brasiliensis [Oryza sativa [Vitis vinifera [tubulins were found and renamed as MeTubulin1 to MeTubulin23 in the order of screening, lower than Hevea brasiliensis (31), but higher than Arabidopsis thaliana (16), Dioscorea rotundata (11), Oryza sativa (16), and Vitis vinifera (18); according to amino acid sequences, motif composition, intron structure, phylogenetic feature, and tubulin type of cassava, the 23 cassava tubulin genes can be divided into four main groups.Tubulins consist of cytoskeleton proteins that are present in all plant species. Understanding the role of tubulins in growth and non-specific response to abiotic stress factors in plants is essential ,51. In othaliana , Dioscorotundata , Hevea biliensis , Oryza sa sativa , and Vitvinifera . In thisMeTubulin23 had the least conserved residues, and MeTubulin2, MeTubulin10, and MeTubulin23 had low similarity with other amino acid sequences  and monocotyledon (Dioscorea rotundata).In the course of evolution, gene diversity proceeds through sequence divergence, recombination, and replication . Gene reMeTubulin in various tissues were analyzed by using RNA-seq (RNA sequencing) data of cassava in the GEO database, which is helpful to research the potential function of the MeTubulin gene . Local BLAST searches were performed based on the hidden Markov model (HMM) profile of tubulin domains from the Pfam database under the accession number PF00091  [https://www.ncbi.nlm.nih.gov/cdd/, access date: 12 July 2020) and SMART website  with default parameters to confirm the conservative tubulin domain [http://web.expasy.org/protparam/, access date: 5 August 2020) and Psort server  with default parameters.The whole genome sequence of cassava was downloaded from the EnsemblPlant database (ly 2020) ,79. The n domain . The seqhttp://gsds.cbi.pku.edu.cn, access date: 21 August 2020) [http://meme.nbcr.net/meme/intro.html, access date: 20 September 2020) was used to analyze the conserved motifs in full-length tubulin proteins, with the following parameters: the site distribution, any number of repetitions; the number of motifs, 10; optimum motif length = 10–20 residues, and shuffle the sequences several times under the same parameters to ensure the reliability of motif [http://www.ebi.ac.uk/Tools/pfa/iprscan/, access date: 8 March 2021).The gene structures were analyzed by the gene structure display server (GSDS: st 2020) . The strof motif . MeanwhiMeTubulin genes on the chromosome was analyzed by MapChart software  [http://circos.ca/, access date: 20 October 2020) [https://sourceforge.net/projects/kakscalculator2/, access date: 18 November 2020) [The location of er 2020) . Multipler 2020) . The syner 2020) . The puter 2020) . Non-syner 2020) .http://plants.ensembl.org/, access date: 22 June 2020). All tubulin proteins were compared by ClustalW [The whole protein sequences of MeTubulin, AtTubulin, HbTubulin, ViTubulin, DrTubulin, and OsTubulin were according to the descriptions in the relevant literature and obtained from the EnsemblPlant database  to research the expression profiles of tubulin genes in different tissue types. Based on Hiplot tool (https://hiplot.com.cn/basic/heatmap access date: 15 February 2021), the absolute FPKM (Fragments per kilobase of exon per million fragments mapped) values were divided by the average of all values, and the ratios were transformed by log2 to obtain data that are suitable for cluster displays, and the heatmap was generated [The RNA-seq data of cassava  were obtenerated .FtsZ2-1 gene under different abiotic stresses and hormone treatments, further qRT-PCR analysis was carried out. For hormone treatments, cassava seedlings were sprayed with 100 μM IAA, MeJA, ABA, and GA3, respectively, and the leaves were collected at 2, 4, 8, 12, 24, and 48 h after treatment. In addition, 300 mM NaCl and 20%PEG were used to simulate salt stress and drought stress, which were consistent with the way of hormone treatments. All treated leaves were immediately frozen in liquid nitrogen and stored at −80 °C for subsequent analysis.SC8 (South China 8), a typical cassava cultivar was derived from the Tropical Crops Genetic Resource Institute . A large number of plants regenerated from tissue culture were obtained by using lateral buds of cassava on MS medium. Then, they were transferred to soil culture for two months to select seedlings with similar growth status for subsequent experiments. To study the expression pattern of the ® Ⅲ SuperMix for qPCR (+ gDNA wiper) Kit (Vazyme #R323) according to the manufacturer’s recommendations. ChamQTM Universal SYBR® qPCR Master Mix (Vazyme #Q711) was used for qRT-PCR on a Roche Lightcyler® 480. The reaction system is as follows: 5.0 μL 2 × ChamQ Universal SYBR ®qPCR Master Mix, 0.2 μL forward primers, 0.2 μL reverse primers, 1.0 μL template cDNA, 3.6 μL ddH2O. The PCR reaction was carried out under the following condition: preincubation (95 °C for 60 s), 40 cycles of three-step amplification , melting , and cooling (37 °C for 30 s). According to the related literature, the cassava β-tubulin gene (MeTubulin1) was suitable to be used as an internal reference for all the qRT-PCR analyses [β-tubulin and FtsZ2-1 (MeTubulin10) CDS sequences . Each sample was performed with three independent replications. Relative expression levels were calculated by the 2−ΔΔCt method [FtsZ2-1 gene at each time point was compared to the corresponding NTC (no treatment control). Statistical differences were analyzed by one-way ANOVA, followed by the post hoc tests, and a p-value less than 0.05 was deemed as significant. The data were processed and plotted in pictures by the GraphPad Prism 7.0.The RNA of each sample was extracted according to Trizol Reagent (Invitrogen #15596026). All RNA was analyzed by 1% agarose gel electrophoresis and then quantified with a Nanodrop ND-1000 spectrophotometer. RNA was used for the synthesis of the first strand of cDNA by using HiScriptanalyses . The speequences  by Primet method . The expMeTubulin genes were identified. The biochemical characteristics of proteins, prediction of subcellular localization, gene structure, conservative motifs, chromosome location, and gene replication were analyzed, and their basic classification and evolutionary characteristics were established. This biological information provides plentiful resources for the functional identification of tubulin genes. The expression atlas analysis of the MeTubulin genes provides evidence of the potential functions of these genes in specific tissues. Moreover, analysis of FtsZ2-1 gene expression under different treatments showed that the FtsZ2-1 gene had a comprehensive response to IAA, MeJA, ABA, NaCl, and PEG. In general, this study laid a foundation for further analysis of the function of the MeTubulin gene family.In the study, a genome-wide analysis of the tubulin family in cassava was carried out, and 23


Genome integrity is constantly threatened by internal and external stressors, in both animals and plants. As plants are sessile, a variety of environment stressors can damage their DNA. In the nucleus, DNA twines around histone proteins to form the higher-order structure “chromatin”. Unraveling how chromatin transforms on sensing genotoxic stress is, thus, key to understanding plant strategies to cope with fluctuating environments. In recent years, accumulating evidence in plant research has suggested that chromatin plays a crucial role in protecting DNA from genotoxic stress in three ways: (1) changes in chromatin modifications around damaged sites enhance DNA repair by providing a scaffold and/or easy access to DNA repair machinery; (2) DNA damage triggers genome-wide alterations in chromatin modifications, globally modulating gene expression required for DNA damage response, such as stem cell death, cell-cycle arrest, and an early onset of endoreplication; and (3) condensed chromatin functions as a physical barrier against genotoxic stressors to protect DNA. In this review, we highlight the chromatin-level control of genome stability and compare the regulatory systems in plants and animals to find out unique mechanisms maintaining genome integrity under genotoxic stress. Arabidopsis thaliana )Chromatin modifications around damaged sites have a significant impact on DNA repair; namely, specific chromatin modifications can function as a scaffold for DNA repair machineries. It is also known that alterations in particular chromatin modifications decrease chromatin compaction, thereby enabling DNA repair proteins to have easy access to the damaged sites.The best-studied histone modification in DDR is phosphorylation of the histone H2A variant H2AX; hereafter, phosphorylated H2AX is referred to as γH2AX. H2AX is phosphorylated at the sites flanking DSBs in an ATM- or ATR-dependent manner, thereby attracting DNA repair proteins ,34,35 F. While γConstitutive heterochromatin is tightly packed and, thus, forms a challenging environment for DNA repair. To repair heterochromatic DSBs, a multistep reaction involving chromatin remodeling is indispensable. Following the induction of DSBs by ionizing radiation, ATM phosphorylates KRAB-ASSOCIATED PROTEIN 1 (KAP1), which induces heterochromatin relaxation, leading to the release of CHROMODOMAIN HELICASE DNA-BINDING PROTEIN 3 (CHD3) from chromatin ,41,42,43Drosophila cells  transcription factors, plays a crucial role in transmitting DNA damage signals in plants ,13,65,66A repair ,69.CBL-INTERACTING PROTEIN KINASE 11 (CIPK11), REPLICATION PROTEIN A 1E (RPA1E), ARGONAUTE 2 (AGO2), and RAD51, all of which are direct targets of SOG1, were elevated significantly after γ-irradiation in Arabidopsis , the former being classified into four types based on amino acid sequence similarities: GNAT (Gcn5-related N-acetyltransferase), p300/CBP, TAFII250, and MYST  families [HAM1 and HAM2 are upregulated upon UV-B irradiation, and that the UV-B-dependent induction of DNA repair genes, such as UV RESISTANCE 2 (UVR2) and UVR7, is compromised in the ham1 or ham2 mutants, compared to wild types, leading to the overaccumulation of CPDs. HAM1 and HAM2 are known to function in globally maintaining H4K5ac [UVR2/7 in UV-B irradiation  and HDA19, in which histone acetylation is increased at the genome-wide level, exhibited a reduced sensitivity to high-B stress [Boron (B) is an essential micronutrient for plant growth and development , but a p factors . Indeed,B stress  (Figure B stress ,89,90; tArabidopsis cultured cells treated with PEO-IAA, an auxin antagonist blocking auxin signaling, were subjected to ATAC-seq analysis, nucleosome-free regions were found to be increased in the genome, especially in the gene body [A possible role of the phytohormone auxin in controlling the chromatin structure was described in suspension cultured cells. Hasegawa et al.  examinedene body . Interesene body . This viene body . Howeverene body ,94, makiene body ,96,97, eArabidopsis root tip, DSBs elevate the cytokinin level and repress the expression of some PIN-FORMED genes encoding auxin efflux carriers, thereby inhibiting downward auxin flow. The resultant reduction of auxin signaling causes cell-cycle arrest in the meristem and stem cell death, which contribute to maintaining genome integrity by providing time for DNA repair and removing DNA-damaged stem cells, respectively [Arabidopsis [Recently, Takahashi et al.  reportedectively . These dectively . In the bidopsis ,101. Thebidopsis .Arabidopsis seedlings can survive 150-Gy γ-irradiation without showing any obvious defects in leaf production [Arabidopsis, as described above, chromatin organization might be one of the crucial factors that determine DNA damage tolerance in plants. Interestingly, a recent study reported that genome size is associated with the sensitivity to DNA damage in plants; plants with a larger genome size are more sensitive to DNA damage than those with smaller genomes [Arabidopsis, which has a smaller genome, does not organize its chromatin into apparent TADs, suggesting that chromosomes can be partitioned into gene-rich euchromatic arms and constitutive heterochromatin [Plants are much more tolerant to genotoxic stress than animals. For instance, 10-Gy irradiation reduces the survival rate of rats to about 20% of those in non-irradiated conditions , whereasoduction . As plan genomes . This fi genomes ,106,107.hromatin . TherefoArabidopsis, which lacks obvious TAD formation, thereby lowering its risk of suffering fatal translocations. This could be one explanation why Arabidopsis can survive higher doses of irradiation than animals. Future studies will precisely estimate the 3D chromatin architecture in plants and animals and identify the cause of DNA damage resistance of plant cells. Additionally, it will be important to explore plant-specific mechanisms to organize higher-order chromosomal architecture, which hormonal signals may control, to illuminate unique strategies of coping with genotoxic stress [Individual chromosomes occupy their own space within the cell nucleus, shaping “chromosome territories”, which are larger in the scale of chromatin compartmentalization than TADs . “Chromac stress ,112,113.Originally, “epigenetics” was a notion proposed from plant research, and various plant-specific epigenetic regulators have been identified so far ,114. Nev