SRFR1 Negatively Regulates Plant NB-LRR Resistance Protein Accumulation to Prevent Autoimmunity

Plant defense responses need to be tightly regulated to prevent auto-immunity, which is detrimental to growth and development. To identify negative regulators of Resistance (R) protein-mediated resistance, we screened for mutants with constitutive defense responses in the npr1-1 background. Map-based cloning revealed that one of the mutant genes encodes a conserved TPR domain-containing protein previously known as SRFR1 (SUPPRESSOR OF rps4-RLD). The constitutive defense responses in the srfr1 mutants in Col-0 background are suppressed by mutations in SNC1, which encodes a TIR-NB-LRR (Toll Interleukin1 Receptor-Nucleotide Binding-Leu-Rich Repeat) R protein. Yeast two-hybrid screens identified SGT1a and SGT1b as interacting proteins of SRFR1. The interactions between SGT1 and SRFR1 were further confirmed by co-immunoprecipitation analysis. In srfr1 mutants, levels of multiple NB-LRR R proteins including SNC1, RPS2 and RPS4 are increased. Increased accumulation of SNC1 is also observed in the sgt1b mutant. Our data suggest that SRFR1 functions together with SGT1 to negatively regulate R protein accumulation, which is required for preventing auto-activation of plant immunity

Optimization of Process Parameters and Analysis of Microstructure and Properties of 18Ni300 by Selective Laser Melting

In this research, we studied the influence of process parameters on the quality of selective laser melting of 18Ni300 maraging steel. The effects of laser power and scanning speed on the relative density and hardness of 18Ni300 were studied by single-factor experiment and the orthogonal experimental method. The relative optimal process parameters of 18Ni300 were obtained when the layer thickness was 0.03 mm, and the hatch space was 0.1 mm. The microstructures and mechanical properties of the samples formed under different process parameters were characterized. The results showed that the optimal hardness and relative density of the sample were 44.7 HRC and 99.98% when the laser power was 230 W and the scanning speed was 1100 mm/s, respectively; the microstructure of the material was uniform and dense, exhibiting few pores. Some columnar crystals appeared along the boundary of the molten pool due to vertical epitaxial growth. The orientation of fine grains at the boundary of the molten pool was random, and some coarse columnar crystals in the molten pool exhibited a certain orientational preference along the <001> orientation. In the case of optimal process parameters, the SLM-formed 18Ni300 was composed of 99.5% martensite and 0.5% retained austenite; the indentation hardness was distributed in the range of 3.2–5 GPa. The indentation modulus was between 142.8–223.4 GPa, exhibiting stronger fluctuations than the indentation hardness. The sample’s mechanical properties showed obvious anisotropy, while the tensile fracture characteristics exhibited necking. The tensile fracture morphology was ductile, and large equiaxed dimples and holes could be observed in the fiber area, accompanied by tearing characteristics

Identification of a novel nonsense mutation in the UNC13D gene from a patient with hemophagocytic lymphohistiocytosis: a case report

Abstract Background Hemophagocytic lymphohistiocytosis (HLH) is a heterogeneous and potentially fatal disease that presents symptoms of persistent fever, splenomegaly and cytopenia. Primary HLH is identified as an autosomal recessive disorder with causative genes including HPLH1, PRF1, UNC13D, STX11 and STXBP2. Case presentation Here, we reported an 8-month-old female patient with compound heterozygosity in the UNC13D gene. The patient, who presented typical symptoms, was diagnosed with HLH based on HLH-2004 guidelines. High-throughput amplicon sequencing for the full-length exon, including a 5 bp padding region and 6 HLH-related genes, was performed to identify the pathogenic mutations in this patient. In all, 9 heterozygous variations were detected, namely, 7 nonpathogenic SNPs, one nonsense mutation (NM_199242.2:c.2206C > T, p.Gln736X), and one splicing mutation (NM_199242.2:c.2709 + 1G > A). These two mutations were considered pathogenic according to previous studies and functional prediction. A two-generation pedigree analysis based on Sanger sequencing was performed to confirm the result. Conclusion Compound heterozygosity in the UNC13D gene was identified in trans and considered a causative mutation in a female patient with HLH. The nonsense mutation (NM_199242.2:c.2206C > T, p.Gln736X) was novel in cases of HLH. Our data expand the spectrum of HLH-related mutations in China and demonstrate the potential of high-throughput amplicon sequencing in the diagnosis of HLH

Gas Expansion Energy Model and Numerical Simulation of Outburst Coal Seam under Multifield Coupling

Due to the insufficient understanding of the outburst mechanism, the coal and gas outburst disasters in China are more serious. Gas expansion energy is the main source of energy that causes outburst. In order to explore the distribution law of gas expansion energy in outburst coal seams, a gas-solid coupling equation of outburst coal seams was established. The distribution law of coal stress field, deformation field, gas flow field, and gas expansion energy were simulated and analyzed by using COMSOL Multiphysics. The results showed that from the excavation face to the deep part of coal seam, the stress presented unloading zone, stress concentration zone, and original stress zone. The volumetric strain and permeability reached the minimum, while the gas pressure reached the maximum at the peak value of vertical stress. As time goes on, the gas pressure in the fracture near the working face gradually decreased and was less than the pressure in coal matrix. The total gas expansion energy consists of free gas and desorption gas expansion energy. Affected by the excavation, free gas expansion energy maintained a constant value in the original coal seam and gradually decreased in the area close to the working face. The expansion energy provided by desorption gas was zero in the original coal seam. And it first increased and then decreased rapidly near the working face. Compared with stress and coal seam thickness, gas pressure and initial diffusion coefficient had significant influence on gas expansion energy of coal seam. When the diffusion coefficient was greater than 1e-9 m2/s, the gas expansion energy of the coal seam near the working face was significantly higher than that of the original coal seam, which had the risk of inducing outburst

Characterization of a novel splicing mutation in UNC13D gene through amplicon sequencing: a case report on HLH

Abstract Background Hemophagocytic lymphohistiocytosis (HLH) is a rare but fatal disease caused by uncontrolled proliferation of activated lymphocytes and macrophages. Six genes including SH2D1A, PRF1, UNC13D, STX11, STXBP2 and XIAP were reported as causative genes in most cases. Case presentation Here we report a novel splicing mutation in UNC13D gene, which was identified in an 18-year-old female. Patient was diagnosed as HLH base on HLH-2004 guidelines, no history of inherited diseases was revealed in this family, parents were healthy and non-consanguineous. Splenomegaly and hemophagocytosis in bone marrow were observed in clinical examination. Amplicon sequencing for the whole coding region of 6 HLH-related genes was performed on Ion S5XL genetic analyzer. In all, four heterozygous mutations were detected, including 2 nonpathogenic SNPs (PRF1:c.900C > T, STX11:c.*70G > A) and 2 splicing mutations in UNC13D gene (UNC13D:c.1299 + 1G > A and UNC13D:c.2709 + 1G > A), both of which were predicted to be potentially pathogenic by human splicing finder (HSF3) tool. The result was confirmed by two-generation pedigree analysis base on sanger sequencing. Conclusions Two compound heterozygous splicing mutations in UNC13D gene were identified and considered to be potential pathogenesis in a female patient of HLH. The mutation UNC13D:c.1299 + 1G > A was reported in HLH for the first time. The inheritance mode and source of the mutation in the proband was examined by family analysis. Our data suggest that further studies of the spectrum of HLH-related mutations in China are warranted

Understanding of Excellent Mechanical Performance of 304L Manufactured by Optimal Selective Laser Melting (SLM) Conditions

The optimal SLM conditions of 304L stainless steel were obtained by single factor and orthogonal tests. Results indicated that the optimal hardness (75 HRB) and Relative Density (RD 99.24%) could be obtained when the laser output power was 190 W, the scanning distance was 0.09 mm and the scanning speed was 800 mm/s. The microstructure of fish scales was uniform and compact with a few pores in the optimal sample. The fine particles were randomly distributed near the edge of the molten pool, and some preferred granular columnar crystal structures were formed. Abundant entanglement dislocations were observed between cell structures, forming dislocation clusters. Spherical nano-precipitates, rich in Si, Mn, and O, were also observed near cell structures. The mechanical properties of the specimens were highly anisotropic, and there were obvious necking and ductility at the tensile fracture

Single-cell combined bioinformatics analysis: construction of immune cluster and risk prognostic model in kidney renal clear cells based on CD8+ T cell-associated genes

Abstract Background Kidney cancer is an immunogenic solid tumor, characterized by high tumor burden and infiltration of CD8+ T cells. Although immunotherapy targeting the PD1/CTLA-4 axis has demonstrated excellent clinical efficacy, clinical outcomes in most patients are poor. Methods We used the RNA sequencing data from the GEO database for KIRC GSE121636 and normal kidney tissue GSE131685, and performed single-cell analysis for cluster identification, pathway enrichment, and CD8+ T cell-associated gene identification. Subsequently, the significance of different CD8+ T-cell associated gene subtypes was elucidated by consensus clustering, pathway analysis, mutated gene analysis, and KIRC immune microenvironment analysis in the TCGA–KIRC disease cohort. Single gene analysis identified LAG3 as the most critical CD8+ T-cell-associated gene and its function was verified by cell phenotype and immunohistochemistry in KIRC. Results In the present study, CD8+ T-cell associated genes in KIRC were screened, including GZMK, CD27, CCL4L2, FXYD2, LAG3, RGS1, CST7, DUSP4, CD8A, and TRBV20-1 and an immunological risk prognostic model was constructed (risk score =  − 0.291858656434841*GZMK − 0.192758342489394*FXYD2 + 0.625023643446193*LAG3 + 0.161324477181591*RGS1 − 0.380169045328895*DUSP4 − 0.107221347575037*TRBV20-1). LAG3 was identified and proved as the most critical CD8+ T cell-associated gene in KIRC. Conclusion We proposed and constructed an immunological risk prognostic model for CD8+ T cell-associated genes and identified LAG3 as a pivotal gene for KIRC progression and CD8+ T-cell infiltration. The model comprehensively explained the immune microenvironment and provided novel immune-related therapeutic targets and biomarkers in KIRC