Modelling the Flow Behaviour of Al Alloy Sheets at Elevated Temperatures Using a Modified Zerilli-Armstrong Model and Phenomenological-Based Constitutive Models.

The flow behaviour of AA2060 Al alloy under warm/hot deformation conditions is complicated because of its dependency on strain rates (ε˙), strain (ε), and deformation modes. Thus, it is crucial to reveal and predict the flow behaviours of this alloy at a wide range of temperatures (T) and ε˙ using different constitutive models. Firstly, the isothermal tensile tests were carried out via a Gleeble-3800 thermomechanical simulator at a T range of 100, 200, 300, 400, and 500 °C and ε˙ range of 0.01, 0.1, 1, and 10 s-1 to reveal the warm/hot flow behaviours of AA2060 alloy sheet. Consequently, three phenomenological-based constitutive models (L-MJC, S1-MJC, S2-MJC) and a modified Zerilli-Armstrong (MZA) model representing physically based constitutive models were developed to precisely predict the flow behaviour of AA2060 alloy sheet under a wide range of T and ε˙. The predictability of the developed constitutive models was assessed and compared using various statistical parameters, including the correlation coefficient (R), average absolute relative error (AARE), and root mean square error (RMSE). By comparing the results determined from these models and those obtained from experimentations, and confirmed by R, AARE, and RMSE values, it is concluded that the predicted stresses determined from the S2-MJC model align closely with the experimental stresses, demonstrating a remarkable fit compared to the S1-MJC, L-MJC, and MZA models. This is because of the linking impact between softening, the strain rate, and strain hardening in the S2-MJC model. It is widely known that the dislocation process is affected by softening and strain rates. This is attributed to the interactions that occurred between ε and ε˙ from one side and between ε, ε˙, and T from the other side using an extensive set of constants correlating the constitutive components of dynamic recovery and softening mechanisms.

Secondary somatosensory cortex glutamatergic innervation of the thalamus facilitates pain.

ABSTRACT: Although the secondary somatosensory cortex (SII) is known to be involved in pain perception, its role in pain modulation and neuropathic pain is yet unknown. In this study, we found that glutamatergic neurons in deep layers of the SII (SII Glu ) responded to bilateral sensory inputs by changing their firing with most being inhibited by contralateral noxious stimulation. Optical inhibition and activation of unilateral SII Glu reduced and enhanced bilateral nociceptive sensitivity, respectively, without affecting mood status. Tracing experiments revealed that SII Glu sent dense monosynaptic projections to the posterolateral nucleus (VPL) and the posterior nucleus (Po) of the thalamus. Optical inhibition and activation of projection terminals of SII Glu in the unilateral VPL and Po inhibited and facilitated pain on the contralateral side, respectively. After partial sciatic nerve ligation, SII Glu became hyperactive as evidenced by higher frequency of spontaneous firing, but the response patterns to peripheral stimulation remained. Optical inhibition of SII Glu alleviated not only bilateral mechanical allodynia and thermal hyperalgesia but also the negative affect associated with spontaneous pain. Inhibition of SII Glu terminals in the VPL and Po also relieved neuropathic pain. This study revealed that SII Glu and the circuits to the VPL and Po constitute a part of the endogenous pain modulatory network. These corticothalamic circuits became hyperactive after peripheral nerve injury, hence contributes to neuropathic pain. These results justify proper inhibition of SII Glu and associated neural circuits as a potential clinical strategy for neuropathic pain treatment.

Coupling Computational Homogenization with Crystal Plasticity Modelling for Predicting the Warm Deformation Behaviour of AA2060-T8 Al-Li Alloy.

This study aimed to propose a new approach for predicting the warm deformation behaviour of AA2060-T8 sheets by coupling computational homogenization (CH) with crystal plasticity (CP) modeling. Firstly, to reveal the warm deformation behaviour of the AA2060-T8 sheet, isothermal warm tensile testing was accomplished using a Gleeble-3800 thermomechanical simulator at the temperatures and strain rates that varied from 373 to 573 K and 0.001 to 0.1 s-1. Then, a novel crystal plasticity model was proposed for describing the grains' behaviour and reflecting the crystals' actual deformation mechanism under warm forming conditions. Afterward, to clarify the in-grain deformation and link the mechanical behaviour of AA2060-T8 with its microstructural state, RVE elements were created to represent the microstructure of AA2060-T8, where several finite elements discretized every grain. A remarkable accordance was observed between the predicted results and their experimental counterparts for all testing conditions. This signifies that coupling CH with CP modelling can successfully determine the warm deformation behaviour of AA2060-T8 (polycrystalline metals) under different working conditions.

Serendipita indica chitinase protects rice from the blast and bakanae diseases.

Serendipita indica, a multifunctional and useful endophyte fungus, has been intensively investigated in promoting plant growth and resistance towards biotic and abiotic stress. Multiple chitinases from microorganisms or plants have been identified to have a high antifungal activity as a biological control. However, chitinase of S. indica still needs to be characterized. We functionally characterized a chitinase (SiChi) in S. indica. The result showed that the purified SiChi protein confers high chitinase activity; importantly, SiChi inhibits the conidial germination of Magnaporthe oryzae and Fusarium moniliforme. After the successful colonization of rice roots by S. indica, both the rice blast disease and bakanae disease were significantly reduced. Interestingly, the purified SiChi could promptly induce rice disease resistance towards M. oryzae and F. moniliforme pathogens when sprayed on rice leaves. Like S. indica, SiChi could upregulate rice pathogen-resistant proteins and defense enzymes. In conclusion, chitinase of S. indica has direct antifungal activity and indirect induced resistance activity, implying an efficient and economic strategy for rice disease control by applying S. indica and SiChi.

Contribution of the Tyrosinase (MoTyr) to Melanin Synthesis, Conidiogenesis, Appressorium Development, and Pathogenicity in Magnaporthe oryzae.

Dihydroxynapthalene-(DHN) and L-dihydroxyphenylalanine (L-DOPA) are two types of dominant melanin in fungi. Fungal melanins with versatile functions are frequently associated with pathogenicity and stress tolerance. In rice blast fungus, Magnaporthe oryzae, DHN melanin is essential to maintain the integrity of the infectious structure, appressoria; but the role of the tyrosinase-derived L-DOPA melanin is still unknown. Here, we have genetically and biologically characterized a tyrosinase gene (MoTyr) in M. oryzae. MoTyr encodes a protein of 719 amino acids that contains the typical CuA and CuB domains of tyrosinase. The deletion mutant of MoTyr (ΔMoTyr) was obtained by using a homologous recombination approach. Phenotypic analysis showed that conidiophore stalks and conidia formation was significantly reduced in ΔMoTyr. Under different concentrations of glycerol and PEG, more appressoria collapsed in the mutant strains than in the wild type, suggesting MoTyr is associated with the integrity of the appressorium wall. Melanin measurement confirmed that MoTyr loss resulted in a significant decrease in melanin synthesis. Accordingly, the loss of MoTyr stunted the conidia germination under stress conditions. Importantly, the MoTyr deletion affected both infection and pathogenesis stages. These results suggest that MoTyr, like DHN pigment synthase, plays a key role in conidiophore stalks formation, appressorium integrity, and pathogenesis of M. oryzae, revealing a potential drug target for blast disease control.

Special-Oriented Annealing-Twins-Induced Orange Peel Morphology of Heat Pipe under Bending Deformation.

The thin-wall heat pipe is an efficient heat transfer component that has been widely used in the field of heat dissipation of high-power electronic equipment in recent years. In this study, the orange peel morphology defect of thin-wall heat pipes after bending deformation was analyzed both for the macro-3D profile and for the micro-formation mechanism. The morphology and crystal orientations of the grains and annealing twins were carefully characterized utilizing optical metallography and the electron backscatter diffraction technique. The results show that after high-temperature sintering treatment, the matrix grains of the heat pipe are seriously coarsened and form a strong Goss texture, while certain annealing twins with the unique copper orientation are retained. The distribution of the Schmid factor value subjected to the uniaxial stress indicates that inhomogeneity in the intergranular deformation exists among the annealing twins and matrix grains. The annealing twin exhibits a "hard-oriented" component during the deformation; thus, it plays a role as a barrier and hinders the slipping of dislocation. As the strain accumulates, part of the annealing twins may protrude from the surface of the heat pipe, forming a large-scale fluctuation of the surface as the so-called "orange peel" morphology. The 3D profile shows the bulged twins mostly perpendicular to the drawing direction, about 200-300 in width and 10-20 µm in height.

Effect of Hot Metal Gas Forming Process on Formability and Microstructure of 6063 Aluminum Alloy Double Wave Tube.

The hot metal gas forming process can significantly improve the formability of a tube and is suitable for the manufacturing of parts with complex shapes. In this paper, a double wave tube component is studied. The effects of different temperatures (400 °C, 425 °C, 450 °C and 475 °C) and different pressures (1 MPa, 1.5 MPa, 2 MPa, 2.5 MPa and 3 MPa) on the formability of 6063 aluminum alloy tubes were studied. The influence of hot metal gas forming process parameters on the microstructure was analyzed. The optimal hot metal gas forming process parameters of 6063 aluminum alloy tubes were explored. The results show that the expansion rate increases with the increase in pressure. The pressure affects the deformation of the tube, which in turn has an effect on the dynamic softening of the material. The expansion rate of parts also increases with the increase in forming temperature. The increased deformation temperature is beneficial to the dynamic recrystallization of 6063, resulting in softening of the material and enhanced deformation uniformity between grains, so that the formability of the material is improved. The optimum hot metal gas forming process parameters of 6063 aluminum alloy tubes are the temperature of 475 °C and the pressure of 2.5 MPa; the maximum expansion ratio is 41.6%.

Bone-Metabolism-Related Serum microRNAs to Diagnose Osteoporosis in Middle-Aged and Elderly Women.

Objective: Postmenopausal osteoporosis (PMOP), a chronic systemic metabolic disease prevalent in middle-aged and elderly women, heavily relies on bone mineral density (BMD) measurement as the diagnostic indicator. In this study, we investigated serum microRNAs (miRNAs) as a possible screening tool for PMOP. Methods: This investigation recruited 83 eligible participants from 795 community-dwelling postmenopausal women between June 2020 and August 2021. The miRNA expression profiles in the serum of PMOP patients were evaluated via miRNA microarray (six PMOP patients and four postmenopausal women without osteoporosis (n-PMOP) as controls). Subsequently, results were verified in independent sample sets (47 PMOP patients and 26 n-PMOP controls) using quantitative real-time PCR. In addition, the target genes and main functions of the differentially expressed miRNAs were explored by bioinformatics analysis. Results: Four highly expressed miRNAs in the serum of patients (hsa-miR-144-5p, hsa-miR-506-3p, hsa-miR-8068, and hsa-miR-6851-3p) showed acceptable disease-independent discrimination performance (area under the curve range: 0.747-0.902) in the training set and verification set, outperforming traditional bone turnover markers. Among four key miRNAs, hsa-miR-144-5p is the only one that can simultaneously predict changes in BMD in lumbar spine 1-4, total hip, and femoral neck (ß = -0.265, p = 0.022; ß = -0.301, p = 0.005; and ß = -0.324, p = 0.003, respectively). Bioinformatics analysis suggested that the differentially expressed miRNAs were targeted mainly to YY1, VIM, and YWHAE genes, which are extensively involved in bone metabolism processes. Conclusions: Bone-metabolism-related serum miRNAs, such as hsa-miR-144-5p, hsa-miR-506-3p, hsa-miR-8068, and hsa-miR-6851-3p, can be used as novel biomarkers for PMOP diagnosis independent of radiological findings and traditional bone turnover markers. Further study of these miRNAs and their target genes may provide new insights into the epigenetic regulatory mechanisms of the onset and progression of the disease.

A Novel Hydroforming Process by Combining Internal and External Pressures for High-Strength Steel Wheel Rims.

As one of the key safety components in motor vehicles, the steel wheel rim is commonly fabricated with the roll forming process. However, due to the varied cross-sections of the rim and the low formability of high-strength steel, it is difficult to produce thin-wall and defect-free wheel rims to realize the purpose of light weight. To solve these problems, a novel hydroforming process by combining internal and external pressures (HIEP) was proposed to produce thin-wall wheel rims in the current study. The designed initial tube with diameter between the maximum and minimum diameter of the wheel rim ensures dispersed deformation and effectively avoids local excessive thinning. During HIEP, a hydroforming process was performed with two successive stages: the external pressure and internal pressure stages. Theoretical analysis and finite element method (FEM) were jointly used to investigate the effect of process parameters on the wrinkling and thinning. With the optimized parameters for internal and external pressure, the wrinkling of wheel rims is prevented under compressive state during the external pressure forming stage. Additionally, HIEP was experimentally carried out with high-strength steel rims of 650 MPa ultimate tensile strength (UTS). Finally, wheel rims with weight reduction of 13% were produced successfully, which shows a uniform thickness distribution with a local maximum thinning ratio of 11.4%.

HMGB1 in the mPFC governs comorbid anxiety in neuropathic pain.

BACKGROUND: Whether neuroinflammation causes comorbid mood disorders in neuropathic pain remains elusive. Here we investigated the role of high mobility group box 1 protein (HMGB1), a proinflammatory cytokine, in the medial prefrontal cortex (mPFC) in anxiety comorbidity of neuropathic pain. METHODS: Neuropathic pain was induced by partial transection of the infraorbital nerve (p-IONX) or partial sciatic nerve ligation (PSL) in mice and evaluated by measuring nociceptive thresholds to mechanical and heat stimulation. Anxiety-like behaviors were assessed by elevated plus maze, light dark box and open field tests. Aversive or anti-aversive effect was detected by conditioned place preference test. Neuronal activity was evaluated by single-unit and patch clamp recordings. The contribution of mPFC pyramidal neurons to anxiety was further examined by selectively inhibiting them by optogenetics. HMGB1 expression was measured by immunohistochemistry and western blotting. Antagonism of HMGB1 was achieved by injecting anti-HMGB1 monoclonal antibody (mAb) intracerebrally or intraperitoneally. RESULTS: Anxiety-like behaviors were presented earlier after p-IONX than after PSL. HMGB1 expression was upregulated in the mPFC temporally in parallel to anxiety onset, rather than in other regions associated with anxiety. The upregulation of HMGB1 expression and its translocation from the nucleus to cytoplasm in the mPFC occurred predominantly in neurons and were accompanied with activation of microglia and astrocytes. Infusion of anti-HMGB1 mAb into the mPFC during the early and late phases after either p-IONX or PSL alleviated anxiety-like behaviors and aversion without changing pain sensitization, while local infusion of exogenous ds-HMGB1, the proinflammatory form of HMGB1, into the mPFC induced anxiety and aversion but not pain sensitization in naïve mice. In addition to reversing established pain sensitization and anxiety simultaneously, intraperitoneal injection of anti-HMGB1 mAb reduced HMGB1 upregulation and suppressed the hyperexcitability of layer 2/3 pyramidal neurons in the mPFC after p-IONX. Moreover, optogenetic inhibition of mPFC pyramidal neurons alleviated anxiety in p-IONX mice. CONCLUSION: These results demonstrate that HMGB1 in the mPFC drives and maintains anxiety comorbidity in neuropathic pain by increasing the excitability of layer 2/3 pyramidal neurons, and justify antagonism of HMGB1, e.g., neutralization by mAb, as a promising therapeutic strategy for neuropathic pain with anxiety comorbidity.

AGC/AKT Protein Kinase SCH9 Is Critical to Pathogenic Development and Overwintering Survival in Magnaporthe oryzae.

Primary inoculum that survives overwintering is one of the key factors that determine the outbreak of plant disease. Pathogenic resting structures, such as chlamydospores, are an ideal inoculum for plant disease. Puzzlingly, Magnaporthe oryzae, a devastating fungal pathogen responsible for blast disease in rice, hardly form any morphologically changed resting structures, and we hypothesize that M. oryzae mainly relies on its physiological alteration to survive overwintering or other harsh environments. However, little progress on research into regulatory genes that facilitate the overwintering of rice blast pathogens has been made so far. Serine threonine protein kinase AGC/AKT, MoSch9, plays an important role in the spore-mediated pathogenesis of M. oryzae. Building on this finding, we discovered that in genetic and biological terms, MoSch9 plays a critical role in conidiophore stalk formation, hyphal-mediated pathogenesis, cold stress tolerance, and overwintering survival of M. oryzae. We discovered that the formation of conidiophore stalks and disease propagation using spores was severely compromised in the mutant strains, whereas hyphal-mediated pathogenesis and the root infection capability of M. oryzae were completely eradicated due to MoSch9 deleted mutants' inability to form an appressorium-like structure. Most importantly, the functional and transcriptomic study of wild-type and MoSch9 mutant strains showed that MoSch9 plays a regulatory role in cold stress tolerance of M. oryzae through the transcription regulation of secondary metabolite synthesis, ATP hydrolyzing, and cell wall integrity proteins during osmotic stress and cold temperatures. From these results, we conclude that MoSch9 is essential for fungal infection-related morphogenesis and overwintering of M. oryzae.

Deformation Characteristics, Formability and Springback Control of Titanium Alloy Sheet at Room Temperature: A Review.

Titanium alloy sheets present inferior formability and severe springback in conventional forming processes at room temperature which greatly restrict their applications in complex-shaped components. In this paper, deformation characteristics and formability and springback behaviors of titanium alloy sheet at room temperature are systematically reviewed. Firstly, deformation characteristics of titanium alloys at room temperature are discussed, and formability improvement under high-rate forming and other methods are summarized, especially the impacting hydroforming developed by us. Then, the main advances in springback prediction and control are outlined, including the advanced constitutive models as well as the optimization of processing paths and parameters. More importantly, notable springback reduction is observed with high strain rate forming methods. Finally, potential investigation prospects for the precise forming of titanium alloy sheet in the future are suggested.

The optimum parameters and neuroimaging mechanism of repetitive transcranial magnetic stimulation to post-stroke cognitive impairment, a protocol of an orthogonally-designed randomized controlled trial.

OBJECTIVE: Repetitive Transcranial Magnetic Stimulation (rTMS) has been used in cognition impairment due to various neuropsychiatric disorders. However, its optimum parameters and the neuroimaging mechanism are still of uncertainty. In order to simulate a study setting as close to real world as possible, the present study introduces a new orthogonally-designed protocol, consisting of the rTMS intervention with four key parameters (stimulating site, frequency, intensity and pulse number) and three different levels in each one, and aims to investigate the optimum parameters and the brain activity and connectivity in default mode network (DMN), dorsal attention network (DAN), central executive network (CEN) following rTMS intervention to post-stroke cognition impairment (PSCI). METHODS: A single-center, orthogonally-designed, triple-blind randomized controlled trial will be conducted and forty-five PSCI patients will be recruited and randomly assigned to one of nine active rTMS groups based on four rTMS paraments: stimulating site, frequency, intensity and pulse number. Neuropsychological, activities of daily living, quality of life and functional magnetic resonance imaging (fMRI) evaluations were be performed pre-, post- and 3 months after rTMS. DISCUSSION: This study evaluates the optimum parameters of rTMS for patients with post-stroke cognition impairment and explores the alteration of neural function in DMN, DAN, CEN brain network. These results would facilitate the standardized application of rTMS in cognition impairment rehabilitation.

Aspartate Transaminase AST2 Involved in Sporulation and Necrotrophic Pathogenesis in the Hemibiotrophs Magnaporthe oryzae and Colletotrichum graminicola.

Aspartate family includes five additional amino acids other than aspartate, among which most except aspartate have been reported for their action in pathogenesis by amino acid biosynthesis. However, how aspartate, the initial substrate of this family metabolic pathway, is involved in pathogenesis remains unknown. Here, we focused on aspartate transaminase (AST) that catalyzes transamination reaction between glutamate-aspartate in Magnaporthe oryzae. Three MoAST genes were bioinformatically analyzed, of which MoAST2 was uniquely upregulated when invasive hyphae switched to necrotrophic pathogenesis. MoAST2 deletion (ΔMoast2) caused a drastic reduction in conidiogenesis and appressorium formation. Particularly, ΔMoast2 was observed to be proliferated at the biotrophic phase but inhibited at the necrotrophic stage, and with invisible symptoms detected, suggesting a critical role in necrotrophic phase. Glutamate family restored the ΔMoast2 defects but aspartate family did not, inferring that transamination occurs from aspartate to glutamine. MoAST2 is cytosolic and possessed H2O2 stress tolerance. In parallel, Colletotrichum graminicola AST2, CgAST2 was proven to be a player in necrotrophic anthracnose development. Therefore, conserved AST2 is qualified to be a drug target for disease control.

Dissecting the Neural Circuitry for Pain Modulation and Chronic Pain: Insights from Optogenetics.

Pain is an unpleasant sensory and emotional experience associated with, or resembling that associated with, actual or potential tissue damage. The processing of pain involves complicated modulation at the levels of the periphery, spinal cord, and brain. The pathogenesis of chronic pain is still not fully understood, which makes the clinical treatment challenging. Optogenetics, which combines optical and genetic technologies, can precisely intervene in the activity of specific groups of neurons and elements of the related circuits. Taking advantage of optogenetics, researchers have achieved a body of new findings that shed light on the cellular and circuit mechanisms of pain transmission, pain modulation, and chronic pain both in the periphery and the central nervous system. In this review, we summarize recent findings in pain research using optogenetic approaches and discuss their significance in understanding the pathogenesis of chronic pain.

Contribution of the Mitochondrial Carbonic Anhydrase (MoCA1) to Conidiogenesis and Pathogenesis in Magnaporthe oryzae.

The interconversion of CO2 and HCO3 - catalyzed by carbonic anhydrases (CAs) is a fundamental biochemical process in organisms. During mammalian-pathogen interaction, both host and pathogen CAs play vital roles in resistance and pathogenesis; during planta-pathogen interaction, however, plant CAs function in host resistance but whether pathogen CAs are involved in pathogenesis is unknown. Here, we biologically characterized the Magnaporthe oryzae CA (MoCA1). Through detecting the DsRED-tagged proteins, we observed the fusion MoCA1 in the mitochondria of M. oryzae. Together with the measurement of CA activity, we confirmed that MoCA1 is a mitochondrial zinc-binding CA. MoCA1 expression, upregulated with H2O2 or NaHCO3 treatment, also showed a drastic upregulation during conidiogenesis and pathogenesis. When MoCA1 was deleted, the mutant ΔMoCA1 was defective in conidiophore development and pathogenicity. 3,3'-Diaminobenzidine (DAB) staining indicated that more H2O2 accumulated in ΔMoCA1; accordingly, ATPase genes were downregulated and ATP content decreased in ΔMoCA1. Summarily, our data proved the involvement of the mitochondrial MoCA1 in conidiogenesis and pathogenesis in the rice blast fungus. Considering the previously reported HCO3 - transporter MoAE4, we propose that MoCA1 in cooperation with MoAE4 constitutes a HCO3 - homeostasis-mediated disease pathway, in which MoCA1 and MoAE4 can be a drug target for disease control.

The Bicarbonate Transporter (MoAE4) Localized on Both Cytomembrane and Tonoplast Promotes Pathogenesis in Magnaporthe oryzae.

Bicarbonate (HCO3-) transporter family including the anion exchanger (AE) group is involved in multiple physiological processes through regulating acid-base homeostasis. HCO3- transporters have been extensively studied in mammals, but fungal homologues of AE are poorly understood. Here, we characterized the AE group member (MoAE4) in Magnaporthe oryzae. MoAE4 exhibits more sequence and structure homologies with the reported AE4 and BOR1 proteins. In addition to the common sublocalization on cytomembrane, MoAE4 also localizes on tonoplast. Yeast complementation verified that MoAE4 rescues boron sensitivity and endows NaHCO3 tolerance in the BOR1 deleted yeast. MoAE4 gene is bicarbonate induced in M. oryzae; and loss of MoAE4 (ΔMoAE4) resulted in mycelial growth inhibited by NaHCO3. Lucigenin fluorescence quenching assay confirmed that ΔMoAE4 accumulated less HCO3- in vacuole and more HCO3- in cytosol, revealing a real role of MoAE4 in bicarbonate transport. ΔMoAE4 was defective in conidiation, appressorium formation, and pathogenicity. More H2O2 was detected to be accumulated in ΔMoAE4 mycelia and infected rice cells. Summarily, our data delineate a cytomembrane and tonoplast located HCO3- transporter, which is required for development and pathogenicity in M. oryzae, and revealing a potential drug target for blast disease control.

A fecal-based test for the detection of advanced adenoma and colorectal cancer: a case-control and screening cohort study.

BACKGROUND: Colorectal cancer (CRC) is the leading cause of cancer death worldwide. Screening is a confirmed way to reduce the incidence and mortality rates of CRC. This study aimed to identify a fecal-based, noninvasive, and accurate method for detection of colorectal cancer (CRC) and advanced adenoma (AA). METHODS: Through detection in tissue (n = 96) and fecal samples (n = 88) and tested in an independent group of fecal samples (n = 294), the methylated DNA marker ITGA4 and bacterial markers Fusobacterium nucleatum (Fn) and Pepetostreptococcusanaerobius (Pa) were identified from the candidate biomarkers for CRC and AA detection. A prediction score (pd-score) was constructed using the selected markers and fecal immunochemical test (FIT) for distinguishing AA and CRC from healthy subjects by logistic regression method. The diagnostic performance of the pd-score was compared with FIT and validated in the external validation cohort (n = 117) and in a large CRC screening cohort. RESULTS: The pd-score accurately identified AA and CRC from healthy subjects with an area under the curve (AUC) of 0.958, at a specificity of 91.37%; the pd-score showed sensitivities of 95.38% for CRC and 70.83% for AA, respectively. In the external validation cohort, the sensitivities of the pd-score for CRC and AA detection were 94.03% and 80.00%, respectively. When applied in screening, the pd-score identified 100% (11/11) of CRC and 70.83% (17/24) of AA in participants with both colonoscopy results and qualified fecal samples, showing an improvement by 41.19% compared to FIT. CONCLUSIONS: The current study developed a noninvasive and well-validated approach for AA and CRC detection, which could be applied widely as a diagnostic and screening test.

Overexpression of an aquaporin protein from Aspergillus glaucus confers salt tolerance in transgenic soybean.

Salt stress is an important abiotic factor that causes severe losses in soybean yield and quality. Therefore, breeding salt-tolerant soybean germplasm resources via genetic engineering has gained importance. Aspergillus glaucus, a halophilic fungus that exhibits significant tolerance to salt, carries the gene AgGlpF. In this study, we used the soybean cotyledonary node transformation method to transfer the AgGlpF gene into the genome of the soybean variety Williams 82 to generate salt-tolerant transgenic soybean varieties. The results of PCR, Southern blot, ddPCR, and RT-PCR indicated that AgGlpF was successfully integrated into the soybean genome and stably expressed. When subjected to salt stress conditions via treatment with 250 mM NaCl for 3 d, the transgenic soybean plants showed significant tolerance compared with wild-type plants, which exhibited withering symptoms and leaf abscission after 9 d. The results of this study indicated that the transfer of AgGlpF into the genome of soybean plants produced transgenic soybean with significantly improved salt stress tolerance.

Alternative Splicing of MoPTEN Is Important for Growth and Pathogenesis in Magnaporthe oryzae.

Human PTEN, a dual-phosphatase tumor suppressor, is frequently dysregulated by alternative splicing. Fungi harbor PTEN homologs, but alternative splicing of fungal PTENs has not been reported as far as we know. Here, we described an alternative splicing case in the PTEN homolog of Magnaporthe oryzae (MoPTEN). Two splice variants of MoPTEN were detected and identified, which are resulted from an intron retention and exclusion (MoPTEN-1/2). Both proteins were different in lipid and protein phosphatase activity and in expression patterns. The MoPTEN deletion mutant (ΔMoPTEN) showed the defects in conidiation, appressorium formation, and pathogenesis. ΔMoPTEN could be completely restored by MoPTEN, but rescued partially by MoPTEN-1 in the defect of conidium and appressorium formation, and by MoPTEN-2 in the defect of invasive development. Assays to assess sensitivity to oxidative stress reveal the involvement of MoPTEN-2 in scavenging exogenous and host-derived H2O2. Taken together, MoPTEN undergoes alternative splicing, and both variants cooperatively contribute to conidium and appressorium development, and invasive hyphae growth in plant cells, revealing a novel disease development pathway in M. oryzae.