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The rice blast fungus Magnaporthe oryzae causes one of the most devastating crop diseases world-wide and new control strategies for blast disease are urgently required. We have used insertional mutagenesis in M. oryzae to define biological processes that are critical for blast disease. Here, we report the identification of LEU2A by T-DNA mutagenesis, which putatively encodes 3-isopropylmalate dehydrogenase (3-IPMDH) required for leucine biosynthesis, implicating that synthesis of this amino acid is required for fungal pathogenesis. M. oryzae contains a further predicted 3-IPMDH gene (LEU2B), two 2-isopropylmalate synthase (2-IPMS) genes (LEU4 and LEU9) and an isopropylmalate isomerase (IPMI) gene (LEU1). Targeted gene deletion mutants of LEU1, LEU2A or LEU4 are leucine auxotrophs, and severely defective in pathogenicity. All phenotypes associated with mutants lacking LEU1, LEU2A or LEU4 could be overcome by adding exogenous leucine. The expression levels of LEU1, LEU2A or LEU4 genes were significantly down-regulated by deletion of the transcription factor gene LEU3, an ortholog of Saccharomyces cerevisiae LEU3. We also functionally characterized leucine biosynthesis genes in the wheat pathogen Fusarium graminearum and found that FgLEU1, FgLEU3 and FgLEU4 are essential for wheat head blight disease, suggesting that leucine biosynthesis in filamentous fungal pathogens may be a conserved factor for fungal pathogenicity and, therefore, a potential target for disease control. [ABSTRACT FROM AUTHOR]

Abstract The North Qaidam ultra-high pressure (UHP) metamorphic belt in the northern Tibetan Plateau is considered as a typical Alpine-type UHP metamorphic belt due to the Early Paleozoic subduction of the Qaidam Block beneath the Qilian Block. The well-preserved Paleozoic metatexite migmatite, diatexite migmatite, felsic sheet/dyke and anatectic granite in the North Qaidam UHP metamorphic belt provide us with an excellent opportunity to study the generation, transport and final fate of crustal melt and geodynamic processes associated with orogenesis. Based on structural relationships, petrology, geochronology, whole-rock geochemistry and Sr Nd isotope data, the Early Paleozoic anatexis during oceanic subduction to continental collision in the North Qaidam UHP metamorphic belt can be further divided into three stages of development ~470 Ma, 446–428 Ma and 432–420 Ma. The first-stage granitic leucosomes are rich in K, poor in Na with low Sr/Y and enrichment of large ion lithophile elements (LILEs), which were probably derived from partial melting of ancient felsic gneiss in continental arc circumstances during oceanic subduction. The second-stage Na-rich leucosomes and tonalite plutons are characterized by high Na, Sr, Sr/Y and La/Yb and low heavy rare earth elements (HREEs), with positive ε Nd (t) values of 0.1–4.3 and zircon ε Hf (t) values of 8.3–10.2, similar to typical tonalite-trondhjemite-granodiorites (TTGs). These TTG-like magmas were produced through partial melting of newly emplaced gabbroic rocks with arc affinity under high-pressure (HP) granulite-facies conditions in a thickened lower crust during continental collision. The volumetrically significant migrated plutons evolved from TTG-like melt will become segments of continental crust and contribute to crustal growth, with partial residual products of HP granulite and/or garnet pyroxenite to the mantle by delamination. The third stage of anatexis was preserved in both migmatitic UHP gneiss and eclogite in the Xitieshan and Luliangshan terranes. The Kfs-rich felsic leucosomes inside UHP gneisses in the third stage are characterized by high alkali contents and low mafic component with FeO T + MgO + TiO 2 <2%. In trace element distribution diagrams, these Kfs-rich leucosomes exhibit parallel patterns to their host gneisses but lower element contents and slightly positive Eu and Sr anomalies. The Pl-rich leucosomes within the retrograde eclogite have geochemical features as follows: (1) rich in CaO, Na 2 O and poor in K 2 O, with Na 2 O/K 2 O ratios >2.0; (2) high La/Yb and Sr/Y, and low Y and HREEs; (3) high Al 2 O 3 and low Mg# values; and (4) enriched in LILEs(e.g., Rb, Ba, K, Sr, Pb) and poor in high field strength elements (HFSEs). Partial melting of UHP eclogite and felsic gneiss during the initial retrogression stage with Pl-rich and Kfs-rich leucosome formation was triggered by dehydration melting involving predominant zoisite and muscovite. The anatectic melts from partial melting of deeply subducted UHP gneiss have accumulated and migrated outside the deeply subducted crustal slice in the form of syn -collisional plutons. The melt evolution from the leucosomes produced at the early exhumation stage to syn-collisional granitoids produced at the late exhumation, might contribute greatly to the exhumation of the North Qaidam UHP metamorphic belt from mantle depths to the lower crustal levels. [ABSTRACT FROM AUTHOR]

[Display omitted] • The dual mechanisms of I 3 –/I– redox and SHES have been proposed to boost cell cycle stability. • The benefits and drawbacks of 15-C-5 on electrolytic liquid system were revealed. • The dual-mechanism electrolyte has more than a twofold boost in cycle life of Li||Li and Li||Cu cells over the blank electrolyte. Uncontrollable lithium dendrite growth and interfacial parasitic reaction hinder the development of lithium metal batteries (LMBs). The 15-Crown-5 (15-C-5) has shown potential to inhibit dendrite growth due to a self-healing electrostatic shielding (SHES) mechanism. The strong complexation of 15-C-5 with Li+ induces to generate a Li 2 O-dominated solid electrolyte interphase (SEI), leading to a dramatic increase in inactive Li and dead Li. Meanwhile, the inhomogeneous SEI causes whisker Li tip deposition, which increases the contact specific surface area between the electrolyte and Li, further intensifies electrolyte consumption. This has impeded the development of 15-C-5 electrolyte. Therefore, we propose to add I 3 –/I– redox to eliminate the negative effect of 15-C-5, including several advantages: the rescuing engineering of inactive Li and dead Li during deposition process; the promotion of columnar Li tips deposition and the circumvention of electrolyte consumption; enhancing SEI mechanical strength and lowering the Li nucleation barrier. The SHES mechanism occurrs during Li deposition and I 3 –/I– redox works during Li stripping. Boosted interfacial stability and cycle life can be retained due to the dual mechanism effect of SHES and I 3 –/I– redox. The work provides a dual-mechanism strategy from optimizing electrolytes to deep the insight into the research toward self-healing lithium metal anodes. [ABSTRACT FROM AUTHOR]

• MoCdc25 plays a key role in mycelium growth, sporulation, conidia morphologenesis and appressorium formation, invasive growth and pathogenicity. • MoCdc25 is involved in regulating the response to osmotic and oxidative stress and cell wall integrity of M. oryzae. • MoCdc25 interacts with Mst50 and acts upstream from MAPK and cAMP-PKA pathways to regulate morphological differentiation and pathogenicity in M. oryzae. Ras guanine nucleotide exchange factors (RasGEFs) can trigger Ras GTPase activities and play important roles in controlling various cellular processes in eukaryotes. Recently, it has been exhibited that RasGEF Cdc25 regulates morphological differentiation and pathogenicity in several plant pathogenic fungi. However, the role of RasGEFs in Magnaporthe oryzae is largely unknown. In this study, we identified and functionally characterized a RasGEF gene MoCDC25 in M. oryzae , which is orthologous to Saccharomyces cerevisiae CDC25. Targeted gene deletion mutants (ΔMocdc25) were completely nonpathogenic and were severely impaired in hyphal growth, conidiation and appressorium formation. The mutants exhibited highly sensitive response to osmotic, cell wall integrity or oxidative stresses. MoCdc25 physically interacts with the MAPK scaffold Mst50 and the putative Cdc42GEF MoScd1 in yeast two-hybrid assays. Moreover, we found that MoCdc25 was involved in regulating the phosphorylation of the MAP kinases (Pmk1, Mps1, and Osm1). In addition, the intracellular cAMP content in hyphae of the ΔMocdc25 mutants was significantly reduced compared to the parent strain Ku80 and the defect of appressorium formation of the mutants could be partially restored by the supplement of exogenous cAMP. Taken together, we conclude that the RasGEF MoCdc25 regulates vegetative growth, conidiation, appressorium formation and pathogenicity via MAPK and cAMP response pathways in M. oryzae. [ABSTRACT FROM AUTHOR]

An edge-rich nitrogen doped micro-mesoporous interconnected carbon nanosphere (N-PCS) was rationally designed and prepared as an electrocatalyst for LSBs. The N-PCS with ultra-high content of edge N atom doping and unique pore structure can effectively bind LiPSs and greatly accelerate their reduction kinetics. As a result, the S/N-PCS operated continuously for more than 1000 cycles at 1.0 C with an ultra-low capacity decay rate (0.04% per cycle), and even provided a high area specific capacity of 5.27 mAh cm−2 at a high sulfur loading of 5.49 mg cm−2. [Display omitted] • We presented an edge-rich nitrogen-doped porous carbon nanospheres with an interconnection structure (N-PCS). • N-PCS possessed ideal structural framework, sophisticated chemical composition and brilliant physical electric-field effect. • Abundant edge doped-nitrogen substances acted as chemisorption centers and catalytic active sites for rapid conversion of LiPSs. • The S/N-PCS operated excellent electrochemical performance even under high sulfur loading and lean electrolyte/sulfur ratio conditions. Theoretically, lithium-sulfur batteries (LSBs) are highly competitive in achieving high energy density storage systems. However, the shuttle effect of polysulfides (LiPSs) and sluggish redox kinetics have severely hampered the commercialization of LSBs. Although a number of catalysts have been applied to solve these problems, their preparation processes and costs are hardly compatible with universalization. Herein, we presented an edge-rich nitrogen-doped porous carbon nanospheres with an interconnection structure (N-PCS) as sulfur redox host, of which mesoporous voids accommodated sulfur to achieve homogenization of sulfur, microporous spaces acted as physical confinement to bind the LiPSs. The interconnected carbon nanosphere could be regarded as penetrated conductive network, which greatly shorten the diffusion distance of Li+ and accelerated carrier diffusion. Moreover, abundant edge doped-nitrogen substances introduced a large number of chemisorption centers and catalytic active sites, realizing the strong adsorption and rapid conversion of LiPSs within the N-PCS skeleton. As a result, the S/N-PCS cathode had a first discharge specific capacity of 755 mAh g−1 at 1.0 C and operated enduringly with an ultra-low capacity decay rate (0.04% per cycle) up to 1000 cycles. Not to be outdone, the S/N-PCS cathode provided area specific capacities of up to 4.76 and 5.27 mAh cm−2 at elevated sulfur loadings of 4.31 and 5.49 mg cm−2. This work not only provides a low-cost improvement strategy, but also expands new research ideas for the development of high-performance LSBs cathodes. [ABSTRACT FROM AUTHOR]

Microglia are immune cells in the central nervous system (CNS) that participate in response to pathological process after ischemic injury. Non-mitogenic fibroblast growth factor 1 (nmFGF1) is an effective neuroprotective factor that is also known as a metabolic regulator. The present study aimed to investigate the effects and mechanism of the neuroprotective ability of nmFGF1 on microglia in mice after photothrombosis (PT) stroke model, to determine whether it could ameliorate ischemic injury in stroke experiment. We discovered that the intranasal administration of nmFGF1 reduced infarct size and ameliorated neurological deficits in behavioral assessment by regulating the secretion of proinflammatory and anti-inflammatory cytokines. Furthermore, in the in vitro experiments, we found that nmFGF1 regulated the expression levels of proinflammatory and anti-inflammatory cytokines in oxygen-glucose deprivation (OGD) and lipopolysaccharide (LPS) stimulation. Evidence have shown that when nuclear factor erythroid 2-related factor 2 (Nfr2) is activated, it inhibits nuclear factor-kappa B (NF-κB) activation to alleviate inflammation. Interestingly, nmFGF1 treatment in vivo remarkably inhibited NF-κB pathway activation and activated Nrf2 pathway. In addition, nmFGF1 and NF-κB inhibitor (BAY11-7082) inhibited NF-κB pathway in LPS-stimulated BV2 microglia. Moreover, in LPS-stimulated BV2 microglia, the anti-inflammatory effect produced by nmFGF1 was knocked down by Nrf2 siRNA. These results indicate that nmFGF1 promoted functional recovery in experimental stroke by modulating microglia/macrophage-mediated neuroinflammation via Nrf2 and NF-κB signaling pathways, making nmFGF1 a potential agent against ischemic stroke. • Intranasal administration of nmFGF1 ameliorated neurological deficits in PT mice. • NmFGF1 influenced microglia polarization process towards M2 phenotype in PT mice. • NmFGF1 alleviated inflammatory response via Nrf2 and NF-κB signaling pathways. [ABSTRACT FROM AUTHOR]

LIM domain proteins contain contiguous double-zinc finger domains and play important roles in cytoskeletal re-organisation and organ development in multi-cellular eukaryotes. Here, we report the characterization of four genes encoding LIM proteins in the rice blast fungus Magnaporthe oryzae. Targeted gene replacement of either the paxillin-encoding gene, PAX1, or LRG1 resulted in a significant reduction in hyphal growth and loss of pathogenicity, while deletion of RGA1 caused defects in conidiogenesis and appressorium development. A fourth LIM domain gene, LDP1, was not required for infection-associated development by M. oryzae. Live cell imaging revealed that Lrg1-GFP and Rga1-GFP both localize to septal pores, while Pax1-GFP is present in the cytoplasm. To explore the function of individual LIM domains, we carried out systematic deletion of each LIM domain, which revealed the importance of the Lrg1-LIM2 and Lrg1-RhoGAP domains for Lrg1 function and overlapping functions of the three LIM domains of Pax1. Interestingly, deletion of either PAX1 or LRG1 led to decreased sensitivity to cell wall-perturbing agents, such as Congo Red and SDS (sodium dodecyl sulfate). qRT-PCR analysis demonstrated the importance of both Lrg1 and Pax1 to regulation of genes associated with cell wall biogenesis. When considered together, our results indicate that LIM domain proteins are key regulators of infection-associated morphogenesis by the rice blast fungus. [ABSTRACT FROM AUTHOR]

Methylenetetrahydrofolate reductases (MTHFRs) play a key role in the biosynthesis of methionine in both prokaryotic and eukaryotic organisms. In this study, we report the identification of a novel T-DNA-tagged mutant WH672 in the rice blast fungus Magnaporthe oryzae, which was defective in vegetative growth, conidiation and pathogenicity. Analysis of the mutation confirmed a single T-DNA insertion upstream of MET13, which encodes a 626-amino-acid protein encoding a MTHFR. Targeted gene deletion of MET13 resulted in mutants that were non-pathogenic and significantly impaired in aerial growth and melanin pigmentation. All phenotypes associated with Δmet13 mutants could be overcome by addition of exogenous methionine. The M. oryzae genome contains a second predicted MTHFR-encoding gene, MET12. The deduced amino acid sequences of Met13 and Met12 share 32% identity. Interestingly, Δmet12 mutants produced significantly less conidia compared with the isogenic wild-type strain and grew very poorly in the absence of methionine, but were fully pathogenic. Deletion of both genes resulted in Δmet13Δmet12 mutants that showed similar phenotypes to single Δmet13 mutants. Taken together, we conclude that the MTHFR gene, MET13, is essential for infection-related morphogenesis by the rice blast fungus M. oryzae. [ABSTRACT FROM AUTHOR]

Background: Postweaning intestinal damage in piglets is a challenging issue in the livestock industry. Short-chain fatty acids (SCFAs) are important metabolic products of the gut microbiota and are widely recognized for their role in maintaining normal colonic function and regulating the intestinal immune system. However, the effects of branched short-chain fatty acid (BSCFA) isobutyrate on intestinal health remain largely unknown. This study aims to explore the potential of isobutyrate for alleviating postweaning intestinal damage., Results: This study indicates that isobutyrate can alleviate diarrhea in weaned piglets, enhance their growth performance, and optimize the gut microbiota. This is mainly achieved through increasing the relative abundance of probiotic bacteria such as Lactobacillus, Megasphaera, and Prevotellaceae&#95;UCG-003, while concurrently reducing the relative abundance of potentially harmful bacteria such as Clostridium&#95;sensu&#95;stricto-1 and Escherichia-Shigella. It promotes the production of SCFAs, including acetate, isobutyrate, and butyrate. Furthermore, it activates G-protein-coupled receptors (GPR43/109A), inhibits the TLR4/MyD88 signaling pathway, strengthens the intestinal barrier function, and regulates the expression of related cytokines., Conclusion: In summary, exogenous isobutyrate can be considered a promising feed additive for improving the intestinal microbiota and regulating intestinal health in piglets. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Chimeric antigen receptor T-cesll therapy (CAR-T) has achieved groundbreaking advancements in clinical application, ushering in a new era for innovative cancer treatment. However, the challenges associated with implementing this novel targeted cell therapy are increasingly significant. Particularly in the clinical management of solid tumors, obstacles such as the immunosuppressive effects of the tumor microenvironment, limited local tumor infiltration capability of CAR-T cells, heterogeneity of tumor targeting antigens, uncertainties surrounding CAR-T quality, control, and clinical adverse reactions have contributed to increased drug resistance and decreased compliance in tumor therapy. These factors have significantly impeded the widespread adoption and utilization of this therapeutic approach. In this paper, we comprehensively analyze recent preclinical and clinical reports on CAR-T therapy while summarizing crucial factors influencing its efficacy. Furthermore, we aim to identify existing solution strategies and explore their current research status. Through this review article, our objective is to broaden perspectives for further exploration into CAR-T therapy strategies and their clinical applications., Competing Interests: The authors declare that they have no competing interests., (© 2024 The Authors.)

Ulcerative colitis (UC) is often accompanied by intestinal inflammation and disruption of intestinal epithelial structures, which are closely associated with changes in the intestinal microbiota. We previously revealed that Min pigs, a native Chinese breed, are more resistant to dextran sulfate sodium (DSS)-induced colitis than commercial Yorkshire pigs. Characterizing the microbiota in Min pigs would allow identification of the core microbes that confer colitis resistance. By analyzing the microbiota linked to the disease course in Min and Yorkshire pigs, we observed that Bacillus spp. were enriched in Min pigs and positively correlated with pathogen resistance. Using targeted screening, we identified and validated Bacillus siamensis MZ16 from Min pigs as a bacterial species with biofilm formation ability, superior salt and pH tolerance, and antimicrobial characteristics. Subsequently, we administered B. siamensis MZ16 to conventional or microbiota-deficient BALB/c mice with DSS-induced colitis to assess its efficacy in alleviating colitis. B. siamensis MZ16 partially counteracted DSS-induced colitis in conventional mice, but it did not mitigate DSS-induced colitis in microbiota-deficient mice. Further analysis revealed that B. siamensis MZ16 administration improved intestinal ecology and integrity and immunological barrier function in mice. Compared to the DSS-treated mice, mice preadministered B. siamensis MZ16 exhibited improved relative abundance of potentially beneficial microbes ( Lactobacillus , Bacillus , Christensenellaceae R7 , Ruminococcus , Clostridium , and Eubacterium ), reduced relative abundance of pathogenic microbes ( Escherichia-Shigella ), and maintained colonic OCLN and ZO-1 levels and IgA and SIgA levels. Furthermore, B. siamensis MZ16 reduced proinflammatory cytokine levels by reversing NF-κB and MAPK pathway activation in the DSS group. Overall, B. siamensis MZ16 from Min pigs had beneficial effects on a colitis mouse model by enhancing intestinal barrier functions and reducing inflammation in a gut microbiota-dependent manner., Competing Interests: Competing interests: The authors declare that they have no competing interests., (Copyright © 2024 Xiuyu Fang et al.)

Breast cancer (BCa) is one of the common malignancies among women. Doxorubicin (Dox), a type of anthracycline anti-tumor drug, is a first-line chemotherapy drug for BCa. It is badly needed to effectively reverse BCa resistance to Dox and improve the clinical symptoms of BCa. Chromatin Modification protein 4C (CHMP4C) is a subunit of the endosomal sorting complex and is expressed in the nucleus and cytoplasm. CHMP4C has been shown to be overexpressed in multiple types of cancers. However, its possible effects on the progression and drug resistance of BCa are still unclear. In this study, we found CHMP4C was highly expressed in BCa tissues and promoted cell proliferation. In addition, CHMP4C promoted resistance of BCa cells to Dox through targeting Snail. We further found that knockdown of CHMP4C inhibited tumor growth and enhanced sensitivity to Dox in vivo. We therefore thought CHMP4C could serve as a target for decreasing BCa drug resistance., (© 2023. The Author(s), under exclusive licence to the Japan Antibiotics Research Association.)

Large amount of coal ash is produced as industrial waste during the electricity generation through the combustion of lignite. Toxic elements arsenic exists in the coal ash, which hinders the subsequent recycling processes. Moreover, coal ash could be recycled further to retrieve scattered metals germanium and tungsten. It is believed that traditional recycling methods present barriers to scaled application, especially serious secondary pollution, such as toxic residue and waste liquid. In this work, a novel sequential vacuum distillation with disulphide method is proposed to separate arsenic, germanium and tungsten from coal ash. First, arsenic can be volatilized completely out of the reaction system at temperatures below 550 °C. Subsequently, Ge and W volatilized in the form of sulfide in the presence of Na 2 S 2 O 3 . The optimal condition was 1050 °C, the mass ratio of 0.6 with reaction a pressure of 1 Pa and a time duration of 120 min demonstrated the best evaporation ratio. For coal fly ash, chemical species As 2 S 3 , GeS, and WO x (x < 3)/WS 2 were the main condensed products. For coal bottom ash, As 2 S 3 , GeS, and WO 3 /WS 2 were dominant chemical components. Mechanisms for the process of release and evaporation of As, Ge, and W from coal ash, vacuum reaction, evaporation, and condensation were analyzed. In summary, the vacuum distillation method deserves to be further developed as it provides an eco-friendly method to recycle coal ash., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests., (Copyright © 2023. Published by Elsevier Ltd.)

Stanniocalcin 1 (Stc1) is well known for its role in regulating calcium uptake in fish by acting on ionocytes or NaR cells. A hallmark of NaR cells is the expression of Trpv6, a constitutively open calcium channel. Recent studies in zebrafish suggest that genetical deletion of Stc1a and Trpv6 individually both increases IGF signaling and NaR cell proliferation. While trpv6 -/- fish suffered from calcium deficiency and died prematurely, stc1a -/- fish had elevated body calcium levels but also died prematurely. The relationship between Stc1a, Trpv6, and IGF signaling in regulating calcium homeostasis and organismal survival is unclear. Here we report that loss of Stc1a increases Trpv6 expression in NaR cells in an IGF signaling-dependent manner. Treatment with CdCl 2 , a Trpv6 inhibitor, reduced NaR cell number in stc1a -/- fish to the sibling levels. Genetic and biochemical analysis results suggest that Stc1a and Trpv6 regulate NaR cell proliferation via the same IGF pathway. Alizarin red staining detected abnormal calcium deposits in the yolk sac region and kidney stone-like structures in stc1a -/- fish. Double knockout or pharmacological inhibition of Trpv6 alleviated these phenotypes, suggesting that Stc1a inhibit epithelial Ca 2+ uptake by regulating Trpv6 expression and activity. stc1a -/- mutant fish developed cardiac edema, body swelling, and died prematurely. Treatment of stc1a -/- fish with CdCl 2 or double knockout of Trpv6 alleviated these phenotypes. These results provide evidence that Stc1a regulates calcium homeostasis and organismal survival by suppressing Trpv6 expression and inhibiting IGF signaling in ionocytes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Li, Li, Wang and Duan.)

Background and Aims: Liver fibrosis predicts adverse clinical outcomes, such as liver-related death (LRD) and hepatocellular carcinoma (HCC) in patients with non-alcoholic fatty liver disease (NAFLD). We aimed to investigate the accuracy of semi-automated quantification of collagen proportionate area (CPA) as an objective new method for predicting clinical outcomes., Method: Liver biopsies from patients with NAFLD underwent computerized image morphometry of Sirius Red staining with CPA quantification performed by ImageScope. Clinical outcomes, including total mortality, LRD, and combined liver outcomes (liver decompensation, HCC, or LRD), were determined by medical records and population-based data-linkage. The accuracy of CPA for predicting outcomes was compared with non-invasive fibrosis tests (Hepascore, FIB-4, APRI)., Results: A total of 295 patients (mean age 50 years) were followed for a median (range) of 9 (0.2-25) years totalling 3253 person-years. Patients with CPA ≥ 10% had significantly higher risks for total death [hazard ratio (HR): 5.0 (1.9-13.2)], LRD [19.0 (2.0-182.0)], and combined liver outcomes [15.6 (3.1-78.6)]. CPA and pathologist fibrosis staging (FS) showed similar accuracy (AUROC) for the prediction of total death (0.68 vs. 0.70), LRD (0.72 vs. 0.77) and combined liver outcomes (0.75 vs. 0.78). Non-invasive serum markers Hepascore, APRI, and FIB-4 reached higher AUROC; however, they were not statistically significant compared to that of CPA except for Hepascore in predicting total mortality (0.86 vs. 0.68, p = 0.009)., Conclusion: Liver fibrosis quantified by CPA analysis was significantly associated with clinical outcomes including total mortality, LRD, and HCC. CPA achieved similar accuracy in predicting outcomes compared to pathologist fibrosis staging and non-invasive serum markers., (© 2023. The Author(s).)

Background and Aims: Management of NAFLD involves noninvasive prediction of fibrosis, which is a surrogate for patient outcomes. We aimed to develop and validate a model predictive of liver-related events (LREs) of decompensation and/or HCC and compare its accuracy with fibrosis models., Approach and Results: Patients with NAFLD from Australia and Spain who were followed for up to 28 years formed derivation (n = 584) and validation (n = 477) cohorts. Competing risk regression and information criteria were used for model development. Accuracy was compared with fibrosis models using time-dependent AUC analysis. During follow-up, LREs occurred in 52 (9%) and 11 (2.3%) patients in derivation and validation cohorts, respectively. Age, type 2 diabetes, albumin, bilirubin, platelet count, and international normalized ratio were independent predictors of LRE and were combined into a model [NAFLD outcomes score (NOS)]. The NOS model calibrated well [calibration slope, 0.99 (derivation), 0.98 (validation)] with excellent overall performance [integrated Brier score, 0.07 (derivation) and 0.01 (validation)]. A cutoff ≥1.3 identified subjects at a higher risk of LRE, (sub-HR 24.6, p < 0.001, 5-year cumulative incidence 38% vs 1.0%, respectively). The predictive accuracy at 5 and 10 years was excellent in both derivation (time-dependent AUC,0.92 and 0.90, respectively) and validation cohorts (time-dependent AUC,0.80 and 0.82, respectively). The NOS was more accurate than the fibrosis-4 or NAFLD fibrosis score for predicting LREs at 5 and 10 years ( p < 0.001)., Conclusions: The NOS model consists of readily available measures and has greater accuracy in predicting outcomes in patients with NAFLD than existing fibrosis models., (Copyright © 2023 American Association for the Study of Liver Diseases.)

Background: Claudin18.2 (CLDN18.2) is a tight junction protein that has been identified as a clinically proven target in gastric cancer. Stimulation of 4-1BB with agonistic antibodies is also a promising strategy for immunotherapy and 4-1BB + T cells were reported to be present within the tumor microenvironment of patients with gastric cancer. However, hepatotoxicity-mediated by 4-1BB activation was observed in clinical trials of agonistic anti-4-1BB monoclonal antibodies., Methods: To specifically activate the 4-1BB + T cells in tumor and avoid the on-target liver toxicity, we developed a novel CLDN18.2×4-1BB bispecific antibody (termed 'givastomig' or 'ABL111'; also known as TJ-CD4B or TJ033721) that was designed to activate 4-1BB signaling in a CLDN18.2 engagement-dependent manner., Results: 4-1BB + T cells were observed to be coexisted with CLDN18.2 + tumor cells in proximity by multiplex immunohistochemical staining of tumor tissues from patients with gastric cancer (n=60). Givastomig/ABL111 could bind to cell lines expressing various levels of CLDN18.2 with a high affinity and induce 4-1BB activation in vitro only in the context of CLDN18.2 binding. The magnitude of T-cell activation by givastomig/ABL111 treatment was closely correlated with the CLDN18.2 expression level of tumor cells from gastric cancer patient-derived xenograft model. Mechanistically, givastomig/ABL111 treatment could upregulate the expression of a panel of pro-inflammatory and interferon-γ-responsive genes in human peripheral blood mononuclear cells when co-cultured with CLDN18.2 + tumor cells. Furthermore, in humanized 4-1BB transgenic mice inoculated with human CLDN18.2-expressing tumor cells, givastomig/ABL111 induced a localized immune activation in tumor as evident by the increased ratio of CD8 + /regulatory T cell, leading to the superior antitumor activity and long-lasting memory response against tumor rechallenge. Givastomig/ABL111 was well tolerated, with no systemic immune response and hepatotoxicity in monkeys., Conclusions: Givastomig/ABL111 is a novel CLDN18.2×4-1BB bispecific antibody which has the potential to treat patients with gastric cancer with a wide range of CLDN18.2 expression level through the restricted activation of 4-1BB + T cells in tumor microenvironment to avoid the risk of liver toxicity and systemic immune response., Competing Interests: Competing interests: ZW, WJ, YZ, ZM, YN, ZS, CC, XL, XC, CL, JZZ, and AXZ are employees of I-Mab Biopharma and JJ, ES, and HC are employees of ABL Bio which develop antibodies presented in this manuscript., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Phosphatidylcholine (PC) plays crucial biological roles in eukaryotic cells. In Saccharomyces cerevisiae , apart from phosphatidylethanolamine (PE) methylation pathway, PC is also synthesized via CDP-choline pathway. Phosphocholine cytidylyltransferase Pct1 is the rate-limiting enzyme to catalyze the conversion from phosphocholine to CDP-choline in this pathway. Here, we report the identification and functional characterization of an ortholog of the budding yeast PCT1 in Magnaporthe oryzae , named MoPCT1 . Targeted gene deletion mutants of MoPCT1 were impaired in vegetative growth, conidiation, appressorium turgor accumulation and cell wall integrity. Also, the mutants were severely compromised in appressorium-mediated penetration, infectious growth and pathogenicity. Western blot analysis revealed that cell autophagy was activated by the deletion of MoPCT1 under nutrient-rich conditions. Moreover, we found several key genes in PE methylation pathway, such as MoCHO2 , MoOPI3 , and MoPSD2 , were significantly up-regulated in the ΔMopct1 mutants, indicating that a pronounced compensation effect exists between the two PC biosynthesis pathways in M. oryzae . Interestingly, in the ΔMopct1 mutants, histone H3 was hypermethylated and expression levels of several methionine cycling-related genes were significantly up-regulated, suggesting that MoPCT1 is involved in histone H3 methylation and methionine metabolism. Taken together, we conclude that the phosphocholine cytidylyltransferase coding gene MoPCT1 plays important roles in vegetative growth, conidiation and appressorium-mediated plant infection by M. oryzae ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Xu, Tong, Lv, Xiao and Wang.)

Background: Oral administration offered a painless way and improved compliance for diabetics. However, the emerging GLP-1 analog peptide drugs for diabetes primarily rely on the injection route, and the development of oral dosage forms was hampered by the low oral bioavailability due to the structural vulnerability to digestive enzymes and molecule impermeability in the gastrointestinal tract., Results: In this study, the non-covalent interaction between cholic acid (CA) and liraglutide (LIRA) was found and theoretically explained by molecular docking simulation. Formation of this physical complex of liraglutide and cholic acid (LIRA/CA Complex) reduced the self-aggregation of LIRA and accelerated intestinal epithelium penetration. By the anti-solvent method, LIRA/CA Complex was loaded into zein/rhamnolipids nanoparticles (LIRA/CA@Zein/RLs) with a loading efficiency of 76.8%. LIRA was protected from fast enzymatic degradation by the hydrophobic zein component. Meanwhile, Rhamnolipids, a glycolipid with surface activity, promoted endocytosis while also stabilizing the nanoparticles. The two components worked synergistically to ensure the delivery of LIRA/CA Complex to intestinal villi and improved oral absorption without disrupting tight junctions. LIRA/CA@Zein/RLs demonstrated a considerable intestinal epithelium absorption in mouse gastrointestinal section and a retention in vivo over 24 h, resulting in a significant and long-lasting hypoglycemic effect in Type 2 diabetes mice., Conclusion: This study provided a promising oral delivery approach for LIRA and exhibited the potential for further translation into clinical application., (© 2023. The Author(s).)

Paxillin is a focal adhesion-associated protein that functions as an adaptor to recruit diverse cytoskeleton and signaling molecules into a complex and plays a crucial role in several signaling pathways in mammal cells. However, paxillin-mediated signal pathways are largely unknown in phytopathogenic fungi. Previously, Pax1 of Magnaporthe oryzae (MoPax1), a paxillin-like protein, has been identified as a crucial pathogenicity determinant. Here, we report the identification of a mitogen-activated protein (MAP) kinase (MAPK) activator, Mka1 of M. oryzae (MoMka1), that physically interacts with MoPax1. Targeted gene deletion of MoMKA1 resulted in pleiotropic defects in aerial hyphal growth, conidiation, appressorium formation, and pathogenicity in M. oryzae. MoMka1 interacts with Mst50, an adaptor protein of the Mst11-Mst7-Pmk1 and Mck1-Mkk2-Mps1 cascades. Moreover, the phosphorylation levels of both Pmk1 and Mps1 in aerial hyphae of the ΔMomka1 mutant were significantly reduced, indicating that MoMka1 acts upstream from the MAPK pathways. Interestingly, we found that MoMka1 interacts with MoAtg6 and MoAtg13. Deletion of MoMKA1 led to impaired MoAtg13 phosphorylation and enhanced autophagic flux under nutrient-rich conditions, indicating that MoMka1 is required for regulation of autophagy in M. oryzae. Taken together, the paxillin MoPax1 may activate MAP kinase signaling pathways and autophagy through MAP kinase activator MoMka1 and play important roles during appressorium-mediated plant infection by the rice blast fungus. IMPORTANCE Paxillin, as an adaptor recruiting diverse cytoskeleton and signaling molecules into a complex, plays a crucial role in several signaling pathways in mammal cells. However, paxillin-mediated signal pathways are largely unknown in phytopathogenic fungi. Here, we identified that MoMka1 physically interacts with MoPax1. Furthermore, MoMka1 acts upstream from the MAPK pathways through interacting with Mst50, a key protein of the Mst11-Mst7-Pmk1 and Mck1-Mkk2-Mps1 cascades. Meanwhile, MoMka1 interacts with both MoAtg6 and MoAtg13 and controls autophagy initiation by influencing the phosphorylation level of MoAtg13. In summary, we describe a model in which MoPax1 activates MAP kinase signaling pathways and autophagy through MoMka1 during appressorium-mediated plant infection by M. oryzae.

Background: Plants can recruit beneficial microbes to enhance their ability to defend against pathogens. However, in contrast to the intensively studied roles of the rhizosphere microbiome in suppressing plant pathogens, the collective community-level change and effect of the phyllosphere microbiome in response to pathogen invasion remains largely elusive., Results: Here, we integrated 16S metabarcoding, shotgun metagenomics and culture-dependent methods to systematically investigate the changes in phyllosphere microbiome between infected and uninfected citrus leaves by Diaporthe citri, a fungal pathogen causing melanose disease worldwide. Multiple microbiome features suggested a shift in phyllosphere microbiome upon D. citri infection, highlighted by the marked reduction of community evenness, the emergence of large numbers of new microbes, and the intense microbial network. We also identified the microbiome features from functional perspectives in infected leaves, such as enriched microbial functions for iron competition and potential antifungal traits, and enriched microbes with beneficial genomic characteristics. Glasshouse experiments demonstrated that several bacteria associated with the microbiome shift could positively affect plant performance under D. citri challenge, with reductions in disease index ranging from 65.7 to 88.4%. Among them, Pantoea asv90 and Methylobacterium asv41 identified as "recruited new microbes" in the infected leaves, exhibited antagonistic activities to D. citri both in vitro and in vivo, including inhibition of spore germination and/or mycelium growth. Sphingomonas spp. presented beneficial genomic characteristics and were found to be the main contributor for the functional enrichment of iron complex outer membrane receptor protein in the infected leaves. Moreover, Sphingomonas asv20 showed a stronger suppression ability against D. citri in iron-deficient conditions than iron-sufficient conditions, suggesting a role of iron competition during their antagonistic action., Conclusions: Overall, our study revealed how phyllosphere microbiomes differed between infected and uninfected citrus leaves by melanose pathogen, and identified potential mechanisms for how the observed microbiome shift might have helped plants cope with pathogen pressure. Our findings provide novel insights into understanding the roles of phyllosphere microbiome responses during pathogen challenge. Video abstract., (© 2022. The Author(s).)

Background:  Hunner's interstitial cystitis (HIC) is a complex disorder characterized by pelvic pain, disrupted urine storage, and Hunner lesions seen on cystoscopy. There are few effective diagnostic biomarkers. In the present study, we used the novel machine learning tool CIBERSORT to measure immune cell subset infiltration and potential novel diagnostic biomarkers for HIC., Methods: The GSE11783 and GSE57560 datasets were downloaded from the Gene Expression Omnibus for analysis. Ten HIC and six healthy samples from GSE11783 were analyzed using the CIBERSORT algorithm. Gene Set Enrichment Analysis (GSEA) was performed to identify biological processes that occur during HIC pathogenesis. Finally, expression levels of 11 T cell follicular helper cell (Tfh) markers were compared between three healthy individuals and four patients from GSE57560., Results: Six types of immune cells in HIC from GSE11783 showed significant differences, including resting mast cells, CD4 + memory-activated T cells (CD3 + CD4 + HLA-DR + cells), M0 and M2 macrophages, Tfh cells, and activated natural killer cells. Except for plasma cells, there were no significant differences between Hunner's lesion and non-Hunner's lesion areas in HIC. The GSEA revealed significantly altered biological processes, including antigen-antibody reactions, autoimmune diseases, and infections of viruses, bacteria, and parasites. There were 11 Tfh cell markers with elevated expression in patients from GSE57560., Conclusion: This was the first demonstration of Tfh cells and CD3 + CD4 + HLA-DR + cells with elevated expression in HIC. These cells might serve as novel diagnostic biomarkers., (© 2021. The Author(s).)

A powder-based 3D printing technology, selective laser sintering (SLS), is a novel strategy of manufacturing complex components with specially tailored properties, including mechanical properties, as well as thermal and electrical conductivity. In this study, the effect of incorporating Al 2 O 3 particles and BN plates on the thermal conductivity of PA12 composites was investigated. PA12 composite powders, which can be well applied to SLS, were prepared via a two-step approach to mixing. Morphology characteristics demonstrated that the fillers dispersed uniformly in the PA12 matrix, as expected. With 35 wt% Al 2 O 3 and 15 wt% BN hybrid fillers, the tensile strength had the potential to reach 25.7 MPa, while the thermal conductivity could reach 1.05 W m -1 K -1 , 275% higher than that of pure PA12. In addition, the study investigated the effects of filler content on the thermal stability and mechanical properties whilst analysing the melting and crystallisation behaviours of SLS components. The results demonstrate that these composites have favourable thermal stability and exhibit no severe deterioration in mechanical properties. The PA12 composites prepared in this work therefore illustrated vast potential in thermal management materials., Competing Interests: The authors declare no conflicts of interest., (This journal is © The Royal Society of Chemistry.)

Background: Increasing evidences indicated that diabetes might increase the incidence of gallbladder cancer. However, no sufficient data has ever clarified the impact of diabetes on the survival of patients with gallbladder cancer., Methods: We comprehensively searched PubMed, Embase, and the Cochrane Library databases through July 2019 in order to find sufficient eligible researches. The pooled hazard risks (HRs) and relative risks (RRs) with 95% confidence intervals (CIs) were calculated with either fix-effects or random-effects model. Due to the low gallbladder cancer mortality in general population, the RRs and standard mortality ratios (SMRs) were considered the similar estimates of the HRs., Results: Ten eligible studies were included in this meta-analysis. Analysis of eight cohorts found that diabetes was closely associated with the mortality of gallbladder cancer (HR = 1.10; 95% CI: 1.06-1.14; P < 0.00001). However, the mortality in male diabetes patients was not higher than female patients (RR = 1.08, 95%CI = 0.57-2.04, P = 0.80)., Conclusions: These findings indicated that diabetes patients had a higher mortality of gallbladder cancer compared with non-diabetes.

Background: In Arabidopsis, a long day flowering plant, CONSTANS (CO) acts as a transcriptional activator of flowering under long day (LD) condition. In rice, a short day flowering plant, Hd1, the ortholog of CO, plays dual functions in respond to day-length, activates flowering in short days and represses flowering in long days. In addition, alleles of Hd1 account for ~ 44% of the variation in flowering time observed in cultivated rice and sorghum. How does it work in bamboo? The function of CO in bamboo is similar to that in Arabidopsis?, Results: Two CO homologous genes, PvCO1 and PvCO2, in Phyllostachys violascens were identified. Alignment analysis showed that the two PvCOLs had the highest sequence similarity to rice Hd1. Both PvCO1 and PvCO2 expressed in specific tissues, mainly in leaf. The PvCO1 gene had low expression before flowering, high expression during the flowering stage, and then declined to low expression again after flowering. In contrast, expression of PvCO2 was low during the flowering stage, but rapidly increased to a high level after flowering. The mRNA levels of both PvCOs exhibited a diurnal rhythm. Both PvCO1 and PvCO2 proteins were localized in nucleus of cells. PvCO1 could interact with PvGF14c protein which belonged to 14-3-3 gene family through B-box domain. Overexpression of PvCO1 in Arabidopsis significantly caused late flowering by reducing the expression of AtFT, whereas, transgenic plants overexpressing PvCO2 showed a similar flowering time with WT under LD conditions. Taken together, these results suggested that PvCO1 was involved in the flowering regulation, and PvCO2 may either not have a role in regulating flowering or act redundantly with other flowering regulators in Arabidopsis. Our data also indicated regulatory divergence between PvCOLs in Ph. violascens and CO in Arabidopsis as well as Hd1 in Oryza sativa. Our results will provide useful information for elucidating the regulatory mechanism of COLs involved in the flowering., Conclusions: Unlike to the CO gene in Arabidopsis, PvCO1 was a negative regulator of flowering in transgenic Arabidopsis under LD condition. It was likely that long period of vegetative growth of this bamboo species was related with the regulation of PvCO1.

This study aimed to elucidate the role of PDIA3 in visceral hypersensitivity of rats with irritable bowel syndrome (IBS). Fourty eight SD rats were randomly divided into four groups (n=12): control group, IBS-empty virus group (IBS-1), IBS-PDIA3 silence group (IBS-2), and IBS-the control group (IBS-3). Visceral hypersensitivity models were established by using acetic acid enema combined with restraint stress, and assayed by abdominal withdrawal reflexes (AWR). Mast cells (MCs) in ileocecal mucosa were counted with toluidine blue staining. Degranulation of MCs was observed under electron microscopy. Serum and mucosal levels of IL-4 and IL-9 were measured with ELISA and QT-PCR. Intestinal tryptase and PAR-2 expression was examined with ELISA and Western blot. The Results showed that PDIA3 plays an important role in the formation of visceral hypersensitivity by increasing systemic and colon mucosal expressions of IL-4 and IL-9, activating mast cells and upregulating PAR-2 expression of target organs., Competing Interests: None., (IJCEP Copyright © 2017.)

Objective: To study the effect of tea saponin in ameliorating nonoxynol(N-9) spermicidal action in vitro., Methods: According to the improved spermicidal test method in vitro recommended by International Planned Parenthood Foundation (IPPF), we evaluated the minimum spermicidal concentration of N-9, tea saponin and their mixed solution in 20 s and 3 min., Results: The minimum spermicidal concentration of N-9 in the mixed solution was (0.13 +/- 0.05) g/L in 20 s and (0.05 +/- 0.004) g/L in 3 min, and that of the tea saponin in the mixed solution was (2.40 +/- 1.07) g/L in 20 s and (1.27 +/- 0.38) g/L in 3 min, compared with the single ingredient N-9 [(0.48 +/- 0.15) g/L in 20 s, (0.34 +/- 0.079 g/L in 3 min], and tea saponin [(5.78 +/- 1.40) g/L in 20 s, (1.71 +/- 0.176) g/L in 3 min], P < 0.01., Conclusion: Tea saponin can improve N-9 spermicidal action in vitro, and tea saponin and nonoxynol have proved of synergic effect.

Objectives: To study the PDE5 activity in corpora cavernosa of the Ganyu Qizhi model penis and the effect of the Chinese herbal medicine Shugan Liqi Huoxue (SLH) ointment on it., Methods: Non-injury stress stimulus method similar to human spirit stress was used to extablish the Ganyu Qizhi animal(rat) model, and the PDE5 activity in corpora cavernosa of the rat penis was measured by the method of immunohistochemistry and computer image analysis., Results: The PDE5 activity in corpora cavernosa of the high-dosage SLH group was significantly different from that of the model group (P < 0.01)., Conclusion: Ganyu Qizhi may increase the PDE5 activity in corpora cavernosa of the penis while SLH can reduce such activity.