Comprehensive analyses of the proteome and ubiquitome revealed mechanism of high temperature accelerating petal abscission in tree peony

Tree peony(Paeonia suffruticosa Andrews)is a well-known ornamental plant with high economic value,but the short fluorescence is a key obstacle to its ornamental value and industry development.High temperature accelerates flower senescence and abscission,but the associated mechanisms are poorly understood.In this study,the tandem mass tag(TMT)proteome and label-free quantitative ubiquitome from tree peony cut flowers treated with 20℃for 0 h(RT0),20℃or 28℃for 60 h(RT60 or HT60)were examined based on morphological observation,respectively.Totally,6970 proteins and 1545 lysine ubiquitinated(Kub)sites in 844 proteins were identified.Hydrophilic residues(such as glutamate and aspartate)neighboring the Kub sites were in preference,and 36.01%of the Kub sites were located on the protein surface.The differentially expressed proteins(DEPs)and Kub-DEPs in HT60 vs RT60 were mainly enriched in ribosomal protein,protein biosynthesis,secondary metabolites biosynthesis,flavonoid metabolism,carbohydrate catabolism,and auxin biosynthesis and signaling revealed by GO and KEGG analysis,accompanying the increase of endogenous abscisic acid(ABA)accumulation and decrease of endogenous indoleacetic acid(IAA)level.Additionally,the expression patterns of six enzymes(SAMS,ACO,YUC,CHS,ANS and PFK)putatively with Kub modifications were analyzed by proteome and real-time quantitative RT-PCR.The cell-free degradation assays showed PsSAMS and PsACO proteins could be degraded via the 26 S proteasome system in tree peony flowers.Finally,a working model was proposed for the acceleration of flower senescence and abscission by high temperature.In summary,all results contributed to understanding the mechanism of flower senescence induced by high temperature and prolonging fluorescence in tree peony.

HBV Infection Promotes the Occurrence and Development of Hepatocellular Carcinoma through Impairing the Inhibitory Effect of PPP2R5A on MAPK/AKT/WNT Signaling Pathway

Reversible phosphorylation and dephosphorylation play important roles in cell function and cell signal transduction. PPP2R5A (protein phosphatase 2 regulatory subunit B’ alpha) is responsible for specifically regulating the catalytic function, substrate specificity and intracellular localization of the tumor suppressor phosphatase PP2A (serine/threonine protein phosphatase 2A). Therefore, the abnormal expression and function of PPP2R5A may be related to canceration. The aim of this study was to reveal its role in the occurrence and development of hepatocellular carcinoma (HCC). It is hoped that the results of this study can provide guidance for the prevention and treatment of HCC. The results showed that PPP2R5A inhibited the proliferation and metastasis of HCC cells, and acted as a tumor suppressor in HCC cells, but it had no significant effect on cell cycle. Further research found that PPP2R5A exerted tumor suppressor efficacy by inhibiting the MAPK/AKT/WNT signaling pathway. Combined with analysis of clinical tissue samples and TCGA database, it was found that the expression of PPP2R5A in tumor tissues of Chinese HCC patients was down-regulated and significantly correlated with the progression-free survival (PFS) of HCC patients. On the contrary, PPP2R5A showed an up-regulation trend in HCC cases in TCGA database although its effect on PFS was the same with that in Chinese HCC patients. Hepatitis B virus (HBV) infection is the main pathogenic factor of HCC in China. It was found that HBV infection reduced the content of PPP2R5A in cells. It was concluded that HBV inhibited the initiation of the protective mechanism mediated by PPP2R5A, making the occurrence and progress of HCC more “unimpeded”. This conclusion will further reveal the role of PPP2R5A in HBV-induced and HBV-unrelated HCC, therefore, providing clues for the prevention and treatment of the two types of HCC, respectively.

Dual functions of PsmiR172b-PsTOE3 module in dormancy release and flowering in tree peony (Paeonia suffruticosa)

MicroRNAs(miRNAs)are non-coding RNAs that interact with target genes and are involved in many physiological processes in plants.miR172-AP2 mainly plays a role in the regulation of f lowering time and floral organ differentiation.Bud dormancy release is necessary for forcing culture of tree peony in winter,but the mechanism of dormancy regulation is unclear.In this study,we found that a miR172 family member,PsmiR172b,was downregulated during chilling-induced bud dormancy release in tree peony,exhibiting a trend opposite to that of PsTOE3.RNA ligase-mediated(RLM)5-RACE(rapid amplification of cDNA ends)confirmed that miR172b targeted PsTOE3,and the cleavage site was between bases 12(T)and 13(C)within the complementary site to miR172b.The functions of miR172b and PsTOE3 were detected by virus-induced gene silencing(VIGS)and their overexpression in tree peony buds.PsmiR172b negatively regulated bud dormancy release,but PsTOE3 promoted bud dormancy release,and the genes associated with bud dormancy release,including PsEBB1,PsEBB3,PsCYCD,and PsBG6,were upregulated.Further analysis indicated that PsTOE3 directly regulated PsEBB1 by binding to its promoter,and the specific binding site was a C-repeat(ACCGAC).Ectopic expression in Arabidopsis revealed that the PsmiR172b-PsTOE3 module displayed conservative function in regulating f lowering.In conclusion,our results provided a novel insight into the functions of PsmiR172-PsTOE3 and possible molecular mechanism underlying bud dormancy release in tree peony.

PsRGL1 negatively regulates chilling- and gibberellin-induced dormancy release by PsF-box1-mediated targeting for proteolytic degradation in tree peony

Tree peony bud endodormancy is a common survival strategy similar to many perennial woody plants in winter,and the activation of the GA signaling pathway is the key to breaking endodormancy.GA signal transduction is involved in many physiological processes.Although the GA-GID1-DELLA regulatory module is conserved in many plants,it has a set of specific components that add complexity to the GA response mechanism.DELLA proteins are key switches in GA signaling.Therefore,there is an urgent need to identify the key DELLA proteins involved in tree peony bud dormancy release.In this study,the prolonged chilling increased the content of endogenously active gibberellins.PsRGL1 among three DELLA proteins was significantly downregulated during chilling-and exogenous GA3-induced bud dormancy release by cell-free degradation assay,and a high level of polyubiquitination was detected.Silencing PsRGL1 accelerated bud dormancy release by increasing the expression of the genes associated with dormancy release,including PsCYCD,PsEBB1,PsEBB3,PsBG6,and PsBG9.Three F-box protein family members responded to chilling and GA3 treatments,resulting in PsF-box1 induction.Yeast two-hybrid and BiFC assays indicated that only PsF-box1 could bind to PsRGL1,and the binding site was in the C-terminal domain.PsF-box1 overexpression promoted dormancy release and upregulated the expression of the dormancy-related genes.In addition,yeast two-hybrid and pull-down assays showed that PsF-box1 also interacted with PsSKP1 to form an E3 ubiquitin ligase.These findings enriched the molecular mechanism of the GA signaling pathway during dormancy release,and enhanced the understanding of tree peony bud endodormancy.

Treatment of Sjogren's Syndrome with Liver Depression

Liu Chunying is a famous old Chinese medicine doctor for more than 30 years.He has rich experience in the understanding and treatment of Sjogren's syndrome(SS).Liu Shi systematically discusses the etiology and pathogenesis of SS from the angle of liver depression.In the treatment,it is emphasized that"Muyu Da Zhi"is the first,with the addition and subtraction of the disease,to provide a new idea for the treatment of Sjogren's syndrome from liver depression,and to attach a test case.

HIV encephalopathy is contributed by macrophages and microglia

Dear Editor,Acquired immunodeficiency syndrome(AIDS),characterized by a significant decrease in the number of CD4^(+)T cells,causes a defective T cell immune response,which subsequently leads to various conditional pathogenic infections and tumors(Calmy et al.,2018;Seligmann et al.,1987).Neurological disorders have been reported in 70%–80%of patients with AIDS(Saylor et al.,2016)。

Broad-spectrum and powerful neutralization of bacterial toxins by erythroliposomes with the help of macrophage uptake and degradation

Anti virulence strategy has been considered as one of the most promising approaches to combat drug-rcesistant bacterial infections.Porc-forming toxins(PFTs)are the largest class of bacterial toxins,inficting their virulence ffect through creating pores on the cell membrane.However,curent solutions for eliminating PFTs are mostly designed based on their molecular structure,requiring customized design for different interactions.In the present study,we employed erythroliposome(denoted as RM-PL),a biomimetic platform constructed by artificial lipid membranes and natural erythrocyle membranes,to necutralize different hemolytic PFTs regardless of their molecular structure.When tested with model PFTs,including a-hemolysin,listeriolysin O,and streptolysin O,RM-PL could completely inhibit toxin-induced hemolysis in a concentration-dependent manner.In vivo studies further confirmed that RM-PL.could eficiently neuralize various toxins and save animals'lives without causing damage to 0rgans or tissues.In addition,we explored the underlying mechanisms of this efficient detoxification ability and found that it was mainly macrophages in the spleen and the liver that took up RM-PL-absorbed toxins through a variety of endocytosis pathways and digested them in lysosomes.In summary,the biomimetic RM-PL presented a promising system for broad-spectrum and powerful toxin neutralization with a mech-anism of lysosome-mediaed loxin degradation.

Morphology and orientation evolution of Cu_(6)Sn_(5)grains on(001)Cu and(011)Cu single crystal substrates under temperature gradient

The morphology and orientation evolution of Cu_(6)Sn_(5)grains formed on(001)Cu and(011)Cu single crystal substrates under temperature gradient(TG)were investigated.The initial orientated prism-type Cu_(6)Sn_(5)grains transformed to non-orientated scallop-type after isothermal reflow.However,the Cu_(6)Sn_(5)grains with strong texture were revealed on cold end single crystal Cu substrates by imposing TG.The Cu_(6)Sn_(5)grains on(001)Cu grew along their c-axis parallel to the substrate and finally merged into one grain to form a fully IMC joint,while those on(011)Cu presented a strong texture and merged into a few dominant Cu_(6)Sn_(5)grains showing about 30°angle with the substrate.The merging between neighboring Cu_(6)Sn_(5)grain pair was attributed to the rapid grain growth and grain boundary migration.Accordingly,a model was put forward to describe the merging process.The different morphology and orientation evolutions of the Cu_(6)Sn_(5)grains on single crystal and polycrystal Cu substrates were revealed based on crystallographic relationship and Cu flux.The method for controlling the morphology and orientation of Cu_(6)Sn_(5)grains is really benefitial to solve the reliability problems caused by anisotropy in 3 D packaging.

Cytokine nanosponges suppressing overactive macrophages and dampening systematic cytokine storm for the treatment of hemophagocytic lymphohistiocytosis

Hemophagocytic lymphohistiocytosis(HLH)is a highly fatal condition with the positive feedback loop between continued immune cell activation and cytokine storm as the core mechanism to mediate multiple organ dysfunction.Inspired by macrophage membranes harbor the receptors with special high affinity for proin-flammation cytokines,lipopolysaccharide(LPS)-stimulated macrophage membrane-coated nanoparticles(LMNP)were developed to show strong sponge ability to both IFN-γand IL-6 and suppressed overactivation of macrophages by inhibiting JAK/STAT signaling pathway both in vitro and in vivo.Besides,LMNP also efficiently alleviated HLH-related symptoms including cytopenia,hepatosplenomegaly and hepatorenal dysfunction and save the life of mouse models.Furthermore,its sponge effect also worked well for five human HLH samples in vitro.Altogether,it’s firstly demonstrated that biocompatible LMNP could dampen HLH with high potential for clinical transformation,which also provided alternative insights for the treatment of other cytokine storm-mediated pathologic conditions such as COVID-19 infection and cytokine releasing syndrome during CAR-T therapy.

黄瓜苋科凝集素基因的表达分析与逆境调控

凝集素是一类具有特异性糖结合活性的蛋白质,通常具有1个或多个非催化的糖结合结构域。凝集素在植物对病原菌的防御反应中发挥重要作用。由于其抗细菌、真菌、病毒和昆虫等的活性,凝集素在农业和生物医药领域都具有很大的应用潜力。作为最小的凝集素家族之一,苋科凝集素的研究较少。该文通过对重要经济作物黄瓜(Cucumissativus)的基因组进行分析,对16种苋科凝集素基因在黄瓜基因组中的分布和位置进行研究,并分析相关基因的外显子/内含子组成。进一步通过启动子分析,阐明了苋科凝集素基因对非生物胁迫的响应情况。最后,通过实时荧光定量PCR,检测了黄瓜中4种苋科凝集素基因对低温、高盐、干旱和ABA处理的响应情况。研究结果可为揭示苋科凝集素的生理功能及其在植物胁迫响应中的作用提供参考。

Shenmai injection enhances cisplatin-induced apoptosis through regulation of Mfn2-dependent mitochondrial dynamics in lung adenocarcinoma A549/DDP cells

Aim:Chemoresistance is the biggest obstacle in cancer treatment.Our previous study demonstrated that Shenmai injection(SMI),a Chinese herbal medicine,enhanced the antitumor effect of cisplatin via glucose metabolism reprogramming.This study aimed to further determine whether the SMI sensitizes the non-small cell lung cancer(NSCLC)cells to cisplatin through regulation mitochondrial dynamics.Methods:The Kaplan-Meier Plotter database was used to investigate the relationship between mRNA expression of mitofusin-2(Mfn2)and the survival analysis of NSCLC patients.The protein expression of Mfn2 in a lung adenocarcinoma tissue chip was detected by immunohistochemistry staining.The expression of Mfn2 and ATAD3A were compared between cisplatin-sensitive A549 and cisplatin-resistant A549/DDP cells.Additionally,A549/DDP cells were co-treated with cisplatin and SMI to detect mitochondrial morphology by fluorescent staining,apoptosis-related protein expression with Western blotting,and mitochondrial membrane potential(ΔΨm)with flow cytometry analysis.Results:The mean survival time of the Mfn2^(low) group was significantly lower than that of the Mfn2^(high) group by Kaplan-Meier Plotter database analysis,and the Mfn2 protein expression level was lower in cancer tissues than in adjacent tissues.The combination of SMI and cisplatin induced dynamic changes in A549/DDP cells,with increased mitochondrial fusion followed by upregulation of Mfn2 and downregulation of ATAD3A and reduced mitochondrial mass and ΔΨm.Moreover,SMI significantly enhanced cisplatin-induced A549/DDP apoptosis,upregulated Bax and the active subunit of caspase-3,and downregulated Bcl-2 expression,as shown via Hoechst staining and flow cytometry analysis.Conclusion:Our findings suggest SMI enhances cisplatin-induced apoptosis through regulation of Mfn2-dependent mitochondrial dynamics in cisplatin-resistant lung adenocarcinoma cells.

Nanoplateletsomes for rapid hemostasis performance

Uncontrollable hemorrhage remains staple trouble in surgical procedures and a leading cause after major trauma.The bleeding issue may trigger various pathologic scenarios that can lead to tissue morbidities and mortalities,and currently available on-site hemostatic agents are confined to a narrow therapeutic index and may carry the risk of immunogenicity.Inspired by the crucial role of platelets in the process of thrombus,a platelet-mimetic plateletsome with wound targeting and blood coagulation properties is developed for hemorrhage control.Plateletsome is formulated by integrating platelet membranes with functionalized synthetic liposomes and exhibits superior wound targeting and effective hemostasis properties.It presents less blood loss and shorter hemostasis time than the platelet membrane vehicles or the conventional liposomes in the mouse tail transection model.The strong homing of the biomimetic plateletsome to the thrombus was also confirmed,demonstrating the potential of this engineered cell membrane vesicle as a biomimetic hemostat for bleeding treatment.

Sqh is involved in the regulation of Ci stability in Hh signaling pathway

Dear Editor,Hedgehog(Hh)pathway is widely involved in cell growth and differentiation during both vertebrate and invertebrate development.Malfunction of Hh pathway couldcausevarioushumandiseases,including birth defects and cancers.In Drosophila,Cubitus interruptus(Ci),the homologue of mammal Glioblastomas(Gli),is a key transcription factor of Hh pathway.The activity andstabilityofCi/Gliare regulatedbyHhconcentration,and further precisely induce downstream gene expressions.Previous studies have suggested that in the absence of Hh,full-length CiFL(Ci155)is partially degraded into the repressor form CiR(Ci75),which blocks downstream gene expressions(Chen and Jiang,2013),through Cul1-Slimb-based E3 ligase-mediated proteolysis(Aza-Blanc et al.,1997;Jia et al.,2002;Price and Kalderon,2002)。