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1.
Biochem Biophys Res Commun ; 710: 149871, 2024 May 28.
Article in English | MEDLINE | ID: mdl-38579538

ABSTRACT

Brassinosteroid activated kinase 1 (BAK1) is a cell-surface coreceptor which plays multiple roles in innate immunity of plants. HopF2 is an effector secreted by the bacterial pathogen Pseudomonas syringae pv. tomato DC3000 into Arabidopsis and suppresses host immune system through interaction with BAK1 as well as its downstream kinase MKK5. The association mechanism of HopF2 to BAK1 remains unclear, which prohibits our understanding and subsequent interfering of their interaction for pathogen management. Herein, we found the kinase domain of BAK1 (BAK1-KD) is sufficient for HopF2 association. With a combination of hydrogen/deuterium exchange mass spectrometry and mutational assays, we found a region of BAK1-KD N-lobe and a region of HopF2 head subdomain are critical for intermolecular interaction, which is also supported by unbiased protein-protein docking with ClusPro and kinase activity assay. Collectively, this research presents the interaction mechanism between Arabidopsis BAK1 and P. syringae HopF2, which could pave the way for bactericide development that blocking the functioning of HopF2 toward BAK1.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Pseudomonas syringae/physiology , Brassinosteroids , Bacterial Proteins/chemistry , Arabidopsis Proteins/physiology , Plant Diseases/microbiology , Protein Serine-Threonine Kinases/chemistry
2.
Nanotechnology ; 35(14)2024 Jan 17.
Article in English | MEDLINE | ID: mdl-38081065

ABSTRACT

The kagome lattice is a well-known model system for the investigation of strong correlation and topological electronic phenomena due to the intrinsic flat band, magnetic frustration, etc. Introducing chirality into the kagome lattice would bring about new physics due to the unique symmetry, which is still yet to be fully explored. Here we report the investigation on a two-dimensional chiral kagome lattice utilizing tight binding band calculation and topological index analysis. It is found that the periodic chiral kagome lattice would bring about a robust zero-energy flat band. Furthermore, in the Su-Schrieffer-Heeger type dimer-/trimerized breathing chiral kagome lattice with particular edge terminations, topological corner states or metallic edge states would appear, implying new candidates for the second-order topological insulator. We also proposed the construction strategy for such lattices employing the scanning tunneling microscope atom manipulation technique.

3.
Proc Natl Acad Sci U S A ; 118(44)2021 11 02.
Article in English | MEDLINE | ID: mdl-34702740

ABSTRACT

Plant nucleotide-binding and leucine-rich repeat (NLR) receptors recognize avirulence effectors directly through their integrated domains (IDs) or indirectly via the effector-targeted proteins. Previous studies have succeeded in generating designer NLR receptors with new recognition profiles by engineering IDs or targeted proteins based on prior knowledge of their interactions with the effectors. However, it is yet a challenge to design a new plant receptor capable of recognizing effectors that function by unknown mechanisms. Several rice NLR immune receptors, including RGA5, possess an integrated heavy metal-associated (HMA) domain that recognizes corresponding Magnaporthe oryzae Avrs and ToxB-like (MAX) effectors in the rice blast fungus. Here, we report a designer rice NLR receptor RGA5HMA2 carrying an engineered, integrated HMA domain (RGA5-HMA2) that can recognize the noncorresponding MAX effector AvrPib and confers the RGA4-dependent resistance to the M. oryzae isolates expressing AvrPib, which originally triggers the Pib-mediated blast resistance via unknown mechanisms. The RGA5-HMA2 domain is contrived based on the high structural similarity of AvrPib with two MAX effectors, AVR-Pia and AVR1-CO39, recognized by cognate RGA5-HMA, the binding interface between AVR1-CO39 and RGA5-HMA, and the distinct surface charge of AvrPib and RAG5-HMA. This work demonstrates that rice NLR receptors with the HMA domain can be engineered to confer resistance to the M. oryzae isolates noncorresponding but structurally similar MAX effectors, which manifest cognate NLR receptor-mediated resistance with unknown mechanisms. Our study also provides a practical approach for developing rice multilines and broad race spectrum-resistant cultivars by introducing a series of engineered NLR receptors.


Subject(s)
NLR Proteins/metabolism , Oryza/genetics , Oryza/immunology , Ascomycota/genetics , Ascomycota/pathogenicity , Disease Resistance/genetics , Fungal Proteins/metabolism , Host-Pathogen Interactions/immunology , NLR Proteins/chemistry , NLR Proteins/genetics , Plant Diseases/microbiology , Plant Proteins/metabolism , Protein Binding , Protein Engineering/methods , Receptors, Immunologic/metabolism
4.
Biochem Biophys Res Commun ; 663: 147-153, 2023 06 30.
Article in English | MEDLINE | ID: mdl-37121125

ABSTRACT

Clathrin-mediated endocytosis (CME) is imperative for physiological processes in eukaryotic cells. In fungi, the Pan1/End3/Sla1 complex controls the transition between early and late stages of CME. Although it is acknowledged that End3 uses its N-terminal to interact with the C-terminal of Sla1, detailed mechanism remains obscure. Magnaporthe oryzae, the pathogenic fungus of rice, cause blast disease that threatens rice production worldwide. Here we report the detailed interaction mechanism between End3 and Sla1 of M. oryzae, i.e. MoEnd3 and MoSla1. The two EH domains of MoEnd3 (MoEnd3-EH1 and MoEnd3-EH2) is different both in evolution and calcium binding, but are indispensable for conformational stability of each other, an unreported effect of tandem-arranged EH domains. MoEnd3-EH1 and MoEnd3-EH2 interact with peptide MoSla11145-1155 containing a NPF motif with a conserved mode, and MoEnd3-EHs (containing both EH1 and EH2 domains) binds MoSla11145-1155 with a higher affinity, supporting the synergetic effect of EH domains. In addition, MoEnd3-EHs also recognize peptide MoSla1971-981 with a new MPF motif that has not been reported before, while Sla1 of yeast contains a DPF motif that bears EH domain interaction ability. Collectively, our research shows that the two EH domains of End3 synergize to interact with dual XPF motifs of Sla1, which conforms to a bivalent receptor-bivalent ligand model to improve both affinity and specificity.


Subject(s)
Peptides , Saccharomyces cerevisiae , Protein Binding , Saccharomyces cerevisiae/metabolism , Peptides/metabolism , Endocytosis/physiology
5.
New Phytol ; 239(6): 2235-2247, 2023 09.
Article in English | MEDLINE | ID: mdl-37403528

ABSTRACT

Heat stress greatly threatens crop production. Plants have evolved multiple adaptive mechanisms, including alternative splicing, that allow them to withstand this stress. However, how alternative splicing contributes to heat stress responses in wheat (Triticum aestivum) is unclear. We reveal that the heat shock transcription factor gene TaHSFA6e is alternatively spliced in response to heat stress. TaHSFA6e generates two major functional transcripts: TaHSFA6e-II and TaHSFA6e-III. TaHSFA6e-III enhances the transcriptional activity of three downstream heat shock protein 70 (TaHSP70) genes to a greater extent than does TaHSFA6e-II. Further investigation reveals that the enhanced transcriptional activity of TaHSFA6e-III is due to a 14-amino acid peptide at its C-terminus, which arises from alternative splicing and is predicted to form an amphipathic helix. Results show that knockout of TaHSFA6e or TaHSP70s increases heat sensitivity in wheat. Moreover, TaHSP70s are localized in stress granule following exposure to heat stress and are involved in regulating stress granule disassembly and translation re-initiation upon stress relief. Polysome profiling analysis confirms that the translational efficiency of stress granule stored mRNAs is lower at the recovery stage in Tahsp70s mutants than in the wild types. Our finding provides insight into the molecular mechanisms by which alternative splicing improves the thermotolerance in wheat.


Subject(s)
Heat-Shock Proteins , Thermotolerance , Heat-Shock Proteins/metabolism , Triticum/metabolism , Alternative Splicing/genetics , Heat-Shock Response/genetics , Thermotolerance/genetics , Gene Expression Regulation, Plant , Plant Proteins/genetics , Plant Proteins/metabolism
6.
J Pineal Res ; 74(1): e12839, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36314656

ABSTRACT

Melatonin is a low-cost natural small indole molecule with versatile biological functions. However, melatonin's fungicidal potential has not been fully exploited, and the mechanism remains elusive. Here, we report that melatonin broadly inhibited 13 plant pathogens. In the rice blast fungal pathogen Magnaporthe oryzae, melatonin inhibited fungal growth, formation of infection-specific structures named appressoria, and plant infection, reducing disease severity. Melatonin entered fungal cells efficiently and colocalized with the critical mitogen-activated protein kinase named Mps1, suppressing phosphorylation of Mps1. Melatonin's affinity for Mps1 via two hydrogen bonds was demonstrated using surface plasmon resonance and chemical modifications. To improve melatonin's efficiency, we obtained 20 melatonin derivatives. Tert-butyloxycarbonyl melatonin showed a 25-fold increase in fungicidal activities, demonstrating the feasibility of chemical modifications in melatonin modification. Our study demonstrated the broad-spectrum fungicidal effect of melatonin by suppressing Mps1 as one of the targets. Through further systematic modifications, developing an eco-friendly melatonin derivative of commercial values for agricultural applications appears promising.


Subject(s)
Melatonin , Oryza , Antifungal Agents/pharmacology , Protein Kinases , Phosphorylation , Plants , Plant Diseases/microbiology
7.
Epidemiol Infect ; 151: e64, 2023 04 03.
Article in English | MEDLINE | ID: mdl-37009679

ABSTRACT

The timely identification of the high-risk groups for nosocomial infections (NIs) plays a vital role in its prevention and control. Therefore, it is crucial to investigate whether the ABO blood group is a risk factor for NI. In this study, patients with NI and non-infection were matched by the propensity score matching method and a logistic regression model was used to analyse the matched datasets. The study found that patients with the B&AB blood group were susceptible to Escherichia coli (OR = 1.783, p = 0.039); the A blood group were susceptible to Staphylococcus aureus (OR = 2.539, p = 0.019) and Pseudomonas aeruginosa (OR = 5.724, p = 0.003); the A&AB blood group were susceptible to Pseudomonas aeruginosa (OR = 4.061, p = 0.008); the AB blood group were vulnerable to urinary tract infection (OR = 13.672, p = 0.019); the B blood group were susceptible to skin and soft tissue infection (OR = 2.418, p = 0.016); and the B&AB blood group were vulnerable to deep incision infection (OR = 4.243, p = 0.043). Summarily, the patient's blood group is vital for identifying high-risk groups for NIs and developing targeted prevention and control measures for NIs.


Subject(s)
ABO Blood-Group System , Cross Infection , Humans , Cross Infection/epidemiology , Escherichia coli , Pseudomonas aeruginosa , Risk Factors , Staphylococcus aureus
8.
Nat Immunol ; 11(10): 905-11, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20802483

ABSTRACT

Interleukin 1ß (IL-1ß) is a key orchestrator of inflammation and host defense that exerts its effects through IL-1 receptor type I (IL-1RI) and IL-1 receptor accessory protein (IL-1RAcP). How IL-1RAcP is recruited by IL-1ß-IL-1RI to form the signaling-competent complex remains elusive. Here we present the crystal structure of IL-1ß bound to IL-1 receptor type II (IL-1RII) and IL-1RAcP. IL-1ß-IL-1RII generated a composite binding surface to recruit IL-1RAcP. Biochemical analysis demonstrated that IL-1ß-IL-1RI and IL-1ß-IL-1RII interacted similarly with IL-1RAcP. It also showed the importance of two loops of IL-1 receptor antagonist (IL-1Ra) in determining its antagonism. Our results provide a structural basis for assembly and activation of the IL-1 receptor and offer a general cytokine-receptor architecture that governs the IL-1 family of cytokines.


Subject(s)
Interleukin-1 Receptor Accessory Protein/chemistry , Interleukin-1beta/chemistry , Receptors, Interleukin-1 Type II/chemistry , Amino Acid Sequence , Crystallography , Humans , Interleukin 1 Receptor Antagonist Protein/chemistry , Interleukin-1 Receptor Accessory Protein/metabolism , Interleukin-1beta/metabolism , Models, Molecular , Molecular Sequence Data , Protein Binding , Protein Structure, Tertiary , Receptors, Interleukin-1 Type II/metabolism , Sequence Alignment , Signal Transduction
9.
J Med Virol ; 94(10): 4975-4982, 2022 10.
Article in English | MEDLINE | ID: mdl-35710693

ABSTRACT

The introduction and scale-up of antiretroviral therapy (ART) have contributed to significantly improved patients with acquired immune deficiency syndrome (AIDS) quality of life and prolongs their survival. This has occurred by suppressing viral replication and recovering the CD4 cell count. However, some patients do not normalize their CD4 cell count, despite suppression of the viral load (VL). Patients with suboptimal immune recovery (SIR), as defined by a VL < 400 copies/ml with a CD4 cell count of<200 cells/µl, after ART initiation, exhibit severe immune dysfunction and have a higher risk of AIDS and non-AIDS events. In recent years, People living with HIV/AIDS (PLWHA) with first-line ART failure began to gradually switch to second-line ART. This study aimed to examine the prevalence and factors affecting SIR among PLWHA who switch to second-line ART in rural China. A 1-year retrospective cohort study was conducted among PLWHA who switched to second-line ART between January 2009 and December 2018. All patients with a VL < 400 copies/ml after 1 year of second-line ART were included. SIR was defined as a CD4 cell count <200 cells/µl and a VL < 400 copies/ml after 1 year of second-line ART. The data collected from medical records were analyzed by univariate and multivariate analyses. A total of 5294 PLWHA met the inclusion criteria, 24 died, and 1152 were lost to follow-up after 1 year of second-line ART. Among 4118 PLWHA who were followed up, 3039 with a VL < 400 copies/ml had their data analyzed, and the prevalence of SIR was 13.1%. The patients' mean age at recruitment was 47.6 ± 8.1 years and 45.3% were men. A total of 30.7% of patients were HIV-positive for >8 years and 88.2% were receiving ART before starting second-line ART for >3 years. The mean CD4 cell count was 354.8 ± 238.2 cells/µl. A multivariable analysis showed that male sex, single status (unmarried or divorced), and a low CD4 cell count were risk factors for SIR among PLWHA with second-line ART. The prevalence of SIR among PLWHA who switched to second-line ART in this retrospective cohort study is lower than that in most other studies. Several factors associated with SIR include male sex, marital status, and CD4 cell count levels in PLWHA.


Subject(s)
Acquired Immunodeficiency Syndrome , Anti-HIV Agents , HIV Infections , Acquired Immunodeficiency Syndrome/drug therapy , Acquired Immunodeficiency Syndrome/epidemiology , Anti-HIV Agents/therapeutic use , Anti-Retroviral Agents/therapeutic use , CD4 Lymphocyte Count , Female , HIV Infections/drug therapy , HIV Infections/epidemiology , Humans , Male , Quality of Life , Retrospective Studies , Viral Load
10.
Phys Rev Lett ; 129(23): 237402, 2022 Dec 02.
Article in English | MEDLINE | ID: mdl-36563194

ABSTRACT

For quasiparticle systems, the control of the quasiparticle lifetime is an important goal, determining whether the related fascinating physics can be revealed in fundamental research and utilized in practical applications. Here, we use double-layer graphene with a boron nitride spacer as a model system to demonstrate that the lifetime of coupled Dirac plasmons can be remotely tuned by electric field-controlled damping pathways. Essentially, one of the graphene layers serves as an external damping amplifier whose efficiency can be controlled by the corresponding doping level. Through this damping switch, the damping rate of the plasmon can be actively tuned up to 1.7 fold. This Letter provides a prototype design to actively control the lifetime of graphene plasmons and also broadens our horizon for the damping control of other quasiparticle systems.

11.
Scand J Gastroenterol ; 57(2): 131-142, 2022 02.
Article in English | MEDLINE | ID: mdl-34738858

ABSTRACT

BACKGROUND AND OBJECTIVES: The association between abdominal obesity and reflux esophagitis (RE) has been extensively evaluated, but the current findings are mixed and more convincing epidemiological evidence urgently needs to be established. To thoroughly explore this relationship, we summarized the latest studies, performed an updated meta-analysis, and examined the dose-response relationship. METHODS: We performed a systematic search of PubMed, Web of Science, and Embase up to 28 March 2021, using prespecified terms to identify studies investigating the association between abdominal obesity and RE. Odds ratios (ORs) with 95% confidence intervals (CIs), mean differences (MDs) or standardized mean differences (SMDs) with 95% CIs were taken as effect-size estimates. RESULTS: Forty-two observational studies, including 11 cohort studies, were meta-analyzed. Overall, a statistically significant association was observed between abdominal obesity and RE, by both the pooled OR (adjusted OR = 1.51, 95% CI: 1.37-1.66, p < .001) and the pooled SMD (SMD = 0.36, 95% CI: 0.30-0.42, p < .001). Moreover, this significant relationship persisted with subgroup stratification. In subgroup analyses, we found that study design, abdominal obesity measurement, adjustment for covariates and sex were possible sources of between-study heterogeneity. For the dose-response analyses, the risk of RE increased with the degree of abdominal obesity, and the increasing trend accelerated when waist circumference (WC) reached 87.0 cm. CONCLUSION: This meta-analysis indicated a significant association between abdominal obesity and RE, and the risk of RE increased with abdominal obesity especially when the WC was over 87.0 cm.


Subject(s)
Esophagitis, Peptic , Obesity, Abdominal , Body Mass Index , Esophagitis, Peptic/epidemiology , Esophagitis, Peptic/etiology , Humans , Obesity, Abdominal/complications , Risk Factors , Waist Circumference/physiology
12.
Sensors (Basel) ; 22(9)2022 Apr 27.
Article in English | MEDLINE | ID: mdl-35591039

ABSTRACT

Object detection is one of the key tasks in an automatic driving system. Aiming to solve the problem of object detection, which cannot meet the detection speed and detection accuracy at the same time, a real-time object detection algorithm (MobileYOLO) is proposed based on YOLOv4. Firstly, the feature extraction network is replaced by introducing the MobileNetv2 network to reduce the number of model parameters; then, part of the standard convolution is replaced by depthwise separable convolution in PAnet and the head network to further reduce the number of model parameters. Finally, by introducing an improved lightweight channel attention modul-Efficient Channel Attention (ECA)-to improve the feature expression ability during feature fusion. The Single-Stage Headless (SSH) context module is introduced to the small object detection branch to increase the receptive field. The experimental results show that the improved algorithm has an accuracy rate of 90.7% on the KITTI data set. Compared with YOLOv4, the parameters of the proposed MobileYOLO model are reduced by 52.11 M, the model size is reduced to one-fifth, and the detection speed is increased by 70%.


Subject(s)
Algorithms , Automobile Driving , Research Design
13.
Sensors (Basel) ; 22(12)2022 Jun 13.
Article in English | MEDLINE | ID: mdl-35746258

ABSTRACT

Contextual information and the dependencies between dimensions is vital in image semantic segmentation. In this paper, we propose a multiple-attention mechanism network (MANet) for semantic segmentation in a very effective and efficient way. Concretely, the contributions are as follows: (1) a novel dual-attention mechanism for capturing feature dependencies in spatial and channel dimensions, where the adjacent position attention captures the dependencies between pixels well; (2) a new cross-dimensional interactive attention feature fusion module, which strengthens the fusion of fine location structure information in low-level features and category semantic information in high-level features. We conduct extensive experiments on semantic segmentation benchmarks including PASCAL VOC 2012 and Cityscapes datasets. Our MANet achieves the mIoU scores of 75.5% and 72.8% on PASCAL VOC 2012 and Cityscapes datasets, respectively. The effectiveness of the network is higher than the previous popular semantic segmentation networks under the same conditions.


Subject(s)
Neural Networks, Computer , Volatile Organic Compounds , Image Processing, Computer-Assisted/methods , Semantics
14.
BMC Infect Dis ; 21(1): 50, 2021 Jan 11.
Article in English | MEDLINE | ID: mdl-33430792

ABSTRACT

BACKGROUND: Hand hygiene (HH) is the cornerstone of infection control, and the promotion of HH is the focus of the world. The study aims to compare the role of two different types of electronic hand hygiene monitoring systems (EHHMSs) in promoting HH of healthcare workers (HCWs) in the intensive care unit (ICU). METHODS: In a 16-bed ICU of a general tertiary hospital in Shenzhen, the research was divided into three stages with interrupted time series (ITS) design. In the first stage, the direct observation method was used to monitor and feed back the HH compliance rate of HCWs monthly. In the second stage, the type1 EHHMS was applied to monitor and feed back the individual number of HH events monthly. In the third stage, the type2 EHHMS with a function of instant reminder and feedback was employed, and the personal HH compliance rates were fed back monthly. Meanwhile, direct observation continued in the last two stages. RESULTS: In the second stage, The HH compliance rate increased. However, there was no significant difference in the trajectory of the rate compared with the first stage. In the first month of the third stage, the HH compliance rate increased by 12.324% immediately and then ascended by 1.242% over time. The number of HH events per bed day and HH products' consumption per bed day were consistent with the change of HH compliance rate observed. CONCLUSION: Monitoring and feedback can improve the HH of HCWs. The EHHMS, with the function of real-time reminders and feedback, has a more noticeable effect on promoting HH.


Subject(s)
Electrical Equipment and Supplies , Hand Hygiene/methods , Health Personnel , Health Promotion/methods , Infection Control/methods , Intensive Care Units , Adult , China , Cross Infection/prevention & control , Feedback , Female , Health Behavior , Humans , Interrupted Time Series Analysis , Male , Reminder Systems , Tertiary Care Centers
15.
Nucleic Acids Res ; 47(8): 4308-4318, 2019 05 07.
Article in English | MEDLINE | ID: mdl-30783673

ABSTRACT

In rice, the critical regulator of the salicylic acid signalling pathway is OsWRKY45, a transcription factor (TF) of the WRKY TF family that functions by binding to the W-box of gene promoters, but the structural basis of OsWRKY45/W-box DNA recognition is unknown. Here, we show the crystal structure of the DNA binding domain of OsWRKY45 (OsWRKY45-DBD, i.e. the WRKY and zinc finger domain) in complex with a W-box DNA. Surprisingly, two OsWRKY45-DBD molecules exchange ß4-ß5 strands to form a dimer. The domain swapping occurs at the hinge region between the ß3 and ß4 strands, and is bridged and stabilized by zinc ion via coordinating residues from different chains. The dimer contains two identical DNA binding domains that interact with the major groove of W-box DNA. In addition to hydrophobic and direct hydrogen bonds, water mediated hydrogen bonds are also involved in base-specific interaction between protein and DNA. Finally, we discussed the cause and consequence of domain swapping of OsWRKY45-DBD, and based on our work and that of previous studies present a detailed mechanism of W-box recognition by WRKY TFs. This work reveals a novel dimerization and DNA-binding mode of WRKY TFs, and an intricate picture of the WRKY/W-box DNA recognition.


Subject(s)
DNA, Plant/chemistry , DNA-Binding Proteins/chemistry , Oryza/genetics , Plant Proteins/chemistry , Protein Subunits/chemistry , Transcription Factors/chemistry , Amino Acid Sequence , Binding Sites , Cloning, Molecular , Crystallography, X-Ray , DNA, Plant/genetics , DNA, Plant/metabolism , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Genetic Vectors/chemistry , Genetic Vectors/metabolism , Models, Molecular , Oryza/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Promoter Regions, Genetic , Protein Binding , Protein Conformation, alpha-Helical , Protein Conformation, beta-Strand , Protein Interaction Domains and Motifs , Protein Multimerization , Protein Subunits/genetics , Protein Subunits/metabolism , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Sequence Alignment , Sequence Homology, Amino Acid , Transcription Factors/genetics , Transcription Factors/metabolism
16.
BMC Public Health ; 21(1): 2022, 2021 11 06.
Article in English | MEDLINE | ID: mdl-34742268

ABSTRACT

BACKGROUND: Prevention and control (P&C) of Corona Virus Disease 2019 (COVID-19) is still a critical task in most countries and regions. However, there are many single evaluation indexes to assess the quality of COVID-19 P&C. It is necessary to synthesize the single evaluation indexes reasonably to obtain the overall evaluation results. METHODS: This study was divided into three steps. Step 1: In February 2020, the improved Delphi method was used to establish the quality evaluation indexes system for COVID-19 P&C. Step 2: in March 2020, the CRITIC method was used to adjust the Order Relation Analysis (G1) method to obtain the subjective and objective (S&O) combination weights. The comprehensive evaluation value was obtained using the weighted Efficacy Coefficient (EC) method, weighted TOPSIS method, weighted rank-sum ratio (RSR) method, and weighted Grey Relationship Analysis (GRA) method. Finally, the linear normalization method was used to synthesize the evaluation values of different evaluation methods. Step 3: From April 2020 to May 2021, this evaluation method was used to monitor and assess COVID-19 P&C quality in critical departments prospectively. The results were reported to the departments monthly. RESULT: A quality evaluation indexes system for COVID-19 P&C was established. Kendall's consistency test shows that the four evaluation method had good consistency (χ2 = 43.429, P<0.001, Kendall's consistency coefficient = 0.835). The Spearman correlation test showed that the correlation between the combined evaluation results and the original method was statistically significant(P < 0.001). According to the Mann-Kendall test, from March 2020 to May 2021, the mean value of COVID-19 P&C quality in all critical departments showed an upward trend (P < 0.01). CONCLUSIONS: The combined comprehensive evaluation method based on the S&O combined weight was more scientific and comprehensive than the single weighting and evaluation methods. In addition, monitoring and feedback of COVID-19 P&C quality were helpful for the improvement of P&C quality.


Subject(s)
COVID-19 , Hospitals, General , Health Services , Humans , Prospective Studies , SARS-CoV-2
17.
Genes Dev ; 27(12): 1339-44, 2013 Jun 15.
Article in English | MEDLINE | ID: mdl-23756652

ABSTRACT

The R-spondin (RSPO) family of secreted proteins (RSPO1-RSPO4) has pleiotropic functions in development and stem cell growth by strongly enhancing Wnt pathway activation. Recently, leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), LGR5, and LGR6 have been identified as receptors for RSPOs. Here we report the complex structure of the LGR4 extracellular domain (ECD) with the RSPO1 N-terminal fragment (RSPO1-2F) containing two adjacent furin-like cysteine-rich domains (FU-CRDs). The LGR4-ECD adopts the anticipated TLR horseshoe structure and uses its concave surface close to the N termini to bind RSPO1-2F. Both the FU-CRD1 and FU-CRD2 domains of RSPO1 contribute to LGR4 interaction, and binding and cellular assays identified critical RSPO1 residues for its biological activities. Our results define the molecular mechanism by which the LGR4/5/6 receptors recognize RSPOs and also provide structural insights into the signaling difference between the LGR4/5/6 receptors and other members in the LGR family.


Subject(s)
Models, Molecular , Thrombospondins/chemistry , Thrombospondins/metabolism , Amino Acid Sequence , Animals , Cell Line , Humans , Molecular Sequence Data , Mutation , Protein Binding , Protein Structure, Tertiary , Receptors, G-Protein-Coupled/chemistry , Receptors, G-Protein-Coupled/genetics , Receptors, G-Protein-Coupled/metabolism , Sequence Alignment , Signal Transduction , Thrombospondins/genetics
18.
J Gene Med ; 22(10): e3240, 2020 10.
Article in English | MEDLINE | ID: mdl-32558063

ABSTRACT

BACKGROUND: Gene therapy has become a potential strategy for cancer treatment. However, the development of efficient gene vectors restricts the application for cancer gene treatment. Functionalization of polymers with functional groups can significantly improve their transfection efficacy. METHODS: Guanidyl can form bidentate hydrogen with the phosphate groups and phosphate groups are present in DNA and cell membranes, thus increasing DNA condensation and cellular uptake. Imidazolyl has high buffering capacity in endosomal/lysosomal acidic environment, facilitating endosome/lysosome escape. We designed a structure-integrated group of guanidyl and imidazolyl, 2-aminoimidazole (AM), which was conjugated to PAMAM generation 2 (G2) for gene therapy of gastric adenocarcinoma. RESULTS: Molecular docking results illustrated that G2-AM bound with DNA molecule effectively via multiple interactions. A quantitative luciferase assay showed that the transfection efficacy of G2-AM/pGL3 was approximately 100-fold greater than that of G2/pGL3, 90-fold greater than that of imidazolyl-modified G2 (G2-M) /pGL3 and 100-fold greater than that of G5/pGL3 without additional cytotoxicity. After introducing the pTRAIL gene into gastric adenocarcinoma cells, the apoptosis ratio of gastric adenocarcinoma cells treated with G2-AM/pTRAIL was 36.95%, which is much larger than the corresponding ratio of G2/pTRAIL (7.45%), G2-M/pTRAIL (11.33%) and G5/pTRAIL (23.2%). In a gastric adenocarcinoma xenograft model, the in vivo transfection efficacy of G2-AM/pRFP was much greater than that of G2/pRFP and G2-M/pRFP. CONCLUSIONS: These results demonstrate that AM could be modified with cationic polymers for potential application in gene delivery and gastric adenocarcinoma gene therapy.


Subject(s)
Adenocarcinoma/therapy , Dendrimers/pharmacology , Genetic Therapy , Stomach Neoplasms/therapy , Adenocarcinoma/genetics , Adenocarcinoma/pathology , Animals , Apoptosis/genetics , Cell Survival/drug effects , Cell Survival/genetics , Genetic Vectors/pharmacology , HEK293 Cells , HeLa Cells , Humans , Mice , Molecular Docking Simulation , Polyamines/pharmacology , Stomach Neoplasms/genetics , Stomach Neoplasms/pathology , Transfection , Xenograft Model Antitumor Assays
19.
Biochem J ; 476(21): 3227-3240, 2019 11 15.
Article in English | MEDLINE | ID: mdl-31455720

ABSTRACT

Trehalose-6-phosphate (T6P) synthase (Tps1) catalyzes the formation of T6P from UDP-glucose (UDPG) (or GDPG, etc.) and glucose-6-phosphate (G6P), and structural basis of this process has not been well studied. MoTps1 (Magnaporthe oryzae Tps1) plays a critical role in carbon and nitrogen metabolism, but its structural information is unknown. Here we present the crystal structures of MoTps1 apo, binary (with UDPG) and ternary (with UDPG/G6P or UDP/T6P) complexes. MoTps1 consists of two modified Rossmann-fold domains and a catalytic center in-between. Unlike Escherichia coli OtsA (EcOtsA, the Tps1 of E. coli), MoTps1 exists as a mixture of monomer, dimer, and oligomer in solution. Inter-chain salt bridges, which are not fully conserved in EcOtsA, play primary roles in MoTps1 oligomerization. Binding of UDPG by MoTps1 C-terminal domain modifies the substrate pocket of MoTps1. In the MoTps1 ternary complex structure, UDP and T6P, the products of UDPG and G6P, are detected, and substantial conformational rearrangements of N-terminal domain, including structural reshuffling (ß3-ß4 loop to α0 helix) and movement of a 'shift region' towards the catalytic centre, are observed. These conformational changes render MoTps1 to a 'closed' state compared with its 'open' state in apo or UDPG complex structures. By solving the EcOtsA apo structure, we confirmed that similar ligand binding induced conformational changes also exist in EcOtsA, although no structural reshuffling involved. Based on our research and previous studies, we present a model for the catalytic process of Tps1. Our research provides novel information on MoTps1, Tps1 family, and structure-based antifungal drug design.


Subject(s)
Fungal Proteins/chemistry , Glucosyltransferases/chemistry , Glucosyltransferases/metabolism , Magnaporthe/enzymology , Biocatalysis , Dimerization , Fungal Proteins/genetics , Fungal Proteins/metabolism , Glucosyltransferases/genetics , Magnaporthe/chemistry , Magnaporthe/genetics , Protein Domains , Uridine Diphosphate Glucose/chemistry , Uridine Diphosphate Glucose/metabolism
20.
Int J Mol Sci ; 21(3)2020 Feb 05.
Article in English | MEDLINE | ID: mdl-32033494

ABSTRACT

High concentrations of Na+ in saline soil impair plant growth and agricultural production. Protein tyrosine phosphorylation is crucial in many cellular regulatory mechanisms. However, regulatory mechanisms of plant protein tyrosine phosphatases (PTPs) in controlling responses to abiotic stress remain limited. We report here the identification of a Tyrosine (Tyr)-specific phosphatase, PdPTP1, from NE19 (Populus nigra × (P. deltoides × P. nigra). Transcript levels of PdPTP1 were upregulated significantly by NaCl treatment and oxidative stress. PdPTP1 was found both in the nucleus and cytoplasm. Under NaCl treatment, transgenic plants overexpressing PdPTP1 (OxPdPTP1) accumulated more Na+ and less K+. In addition, OxPdPTP1 poplars accumulated more H2O2 and O2·-, which is consistent with the downregulation of enzymatic ROS-scavengers activity. Furthermore, PdPTP1 interacted with PdMAPK3/6 in vivo and in vitro. In conclusion, our findings demonstrate that PdPTP1 functions as a negative regulator of salt tolerance via a mechanism of affecting Na+/K+ and ROS homeostasis.


Subject(s)
Homeostasis/genetics , Plant Proteins/genetics , Populus/genetics , Protein Tyrosine Phosphatases/genetics , Reactive Oxygen Species/metabolism , Salt Tolerance/genetics , Sodium/metabolism , Amino Acid Sequence , Down-Regulation/genetics , Gene Expression Regulation, Plant/genetics , Hydrogen Peroxide/metabolism , Oxidative Stress/genetics , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Populus/metabolism , Potassium/metabolism , Sequence Alignment , Stress, Physiological/genetics , Up-Regulation/genetics
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