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Abstract Background Breast cancer (BC) is the most prevalent malignant tumor among women worldwide and a significant cause of cancer-related deaths in females. Recent studies have shown that lipid metabolism-related genes (LMRGs) exhibit prognostic potential in various types of tumors, including BC. Our study aimed to establish a novel model to predict the metastasis of BC. Methods Clinical information and corresponding RNA data of patients with BC were downloaded from The Cancer Genome Atlas and Gene Expression Omnibus databases. Consensus clustering was performed to identify novel molecular subgroups. Estimation of Stromal and Immune Cells in Malignant Tumor Tissues using Expression, microenvironment cell populations counter, microenvironment cell populations counter, and single-sample gene set enrichment analyses were employed to determine the tumor immune microenvironment and immune status of the identified subgroups. Functional analyses, including Gene Ontology and gene set enrichment analyses, were conducted to elucidate the underlying mechanisms. A prognostic risk model was constructed using the Least Absolute Shrinkage and Selection Operator algorithm and multivariate Cox regression analysis. Results This study identified differential gene expression between patients with BC exhibiting metastasis and those without metastasis using public databases. Using the obtained data, we established predictive models based on six LMRGs. Furthermore, consensus clustering and prognostic score grouping analysis revealed that differentially expressed LMRGs influence tumor prognosis by regulating tumor immunity. To facilitate clinical application, we developed a nomogram integrating the risk model and clinical characteristics to accurately predict the prognosis of patients with BC. Conclusion We developed and validated a novel signature associated with LMRGs for predicting disease-free survival in patients with BC. The expression of LMRGs correlates with the immune microenvironment of patients with BC, providing new insights and improved strategies for the diagnosis and treatment of BC.

Abstract Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a devastating disease that seriously threatens the production of crops worldwide. Triadimefon is the widely-used fungicide for controlling the disease in China; however, as the fungicide targets a single site (position 401 in the 134th codon of the Cyp51 gene), the extensive application imposes a strong selection pressure on the pathogens, which may potentially lose the effect over time. In this study, 176 Pst field isolates sampled from different regions of Xinjiang were determined for their sensitivity to triadimefon because it is the few frequent Pst outbreak and representative area in China. The results showed that the Pst isolates collected from Yili, Xinjiang, exhibited a strong resistance to triadimefon with an average EC50 of 0.263 µg/mL, despite the rest of the isolates maintaining high sensitivity to triadimefon. The triadimefon-resistant and triadimefon-sensitive isolates did not display significant differences in sporulation, but the triadimefon-resistant isolates exhibited weaker adaptive traits in their latent period and urediniospore germination rate than the triadimefon-sensitive isolates. No cross-resistance was found for the other two fungicides, flubeneteram or pyraclostrobin; however, cross-resistance for the demethylation inhibitor (DMI) fungicides, tebuconazole and hexaconazole, was found. Genome sequencing revealed that the Tyrosine (Y) at 134 residue was mutated to Phenylalanine (F) in the Xinjiang isolates. Our study revealed that a natural mutation in Pst led to the efficacy loss of triadimefon to control the disease.

Objectives: This study aimed to describe the effects of low-dose (prednisolone acetate 2.5–7.5 mg/day) glucocorticoids (GCs) maintenance therapy in patients with primary nephrotic syndrome (NS) suffering from coronavirus disease 2019 (COVID-19).Methods: A single-center retrospective study of NS patients with COVID-19 infection in Zhongda Hospital Affiliated to Southeast University from 1 February 2022 to 31 March 2023 was conducted. All enrolled patients underwent renal biopsy for the pathological diagnosis and reached complete remission (CR) or near-CR before COVID-19 infection. According to the maintained therapy regimen, patients were divided into low-dose GCs group and non-GCs group.Results: A total of 125 patients were enrolled in the study. Their median age was 46.0 ± 15.6 years, and the median value of 24-h urine protein was 0.77 g. The majority of these patients received treatment for more than 6 months, with a significant portion achieving CR (29.6%) or near-CR (43.2%). The leading cause of NS was membranous nephropathy (52%). There were no significant differences in the baseline characteristics between low-dose GCs and non-GCs group. As compared to those in the non-GCs group, patients receiving low-dose GCs treatment showed less fatigue or muscle weakness, smell disorder, palpitations, decreased appetite, taste disorder, dizziness, sore throat or difficult to swallow and fever (p < 0.05). Moreover, patients in the low-dose GCs group were with higher median quality of life scores (85.0) than in the non-GCs group (p = 0.001). Further serum inflammatory factor analysis indicated that interleukin-6 (IL-6) levels in the non-GCs group were significantly higher than that in the low-dose GCs group (p < 0.05).Conclusion: Patients with NS in low-dose GCs maintenance therapy stage showed milder symptom, higher quality of life and decreased serum IL-6 levels compared to those, who were not on GCs maintenance therapy. These results suggest the beneficial effect of low-dose GCs therapy in NS patients with CR/near-CR suffering from COVID-19 infection.

Objective To explore the mechanism of FGFR3⁃TACC3 (F3⁃T3) fusion gene mediating DNA damage repair through promoting pyruvate kinase M2 (PKM2)'s nuclear translocation in glioblastoma. Methods The lentiviral transfection technology was used to constructed stable transitional cell lines U87MG cells and U251MG cells that stably expressing F3 ⁃ T3 and empty vector. In vivo glioblastoma model was constructed by intracranial in situ tumor implantation in athymic mice, and the tumorigenic ability of F3⁃T3 transfected cells in athymic mice of each treatment group was observed by small⁃animal in vivo imaging system. Bioinformatics analysis was performed to analyze gene microarray data exploring the possible biological functions of F3 ⁃ T3 mediating chemoresistance and identify the relationship between survival expectations and PKM2 expression levels in glioblastoma patients from The Cancer Genome Atlas (TCGA) database. Transient transfection of small interference RNA (siRNA) was used to knock down the expression of PKM2 in the F3 ⁃ T3 transfected cells. Proliferative activity of U87MG and U251MG cells treated with different concentrations of temozolomide (TMZ), transfected with siRNA, and TMZ in combination with Compound 3k, a PKM2 inhibitor, was observed in CCK ⁃ 8 cell proliferation assays. Nuclear and cytoplasmic proteins were extracted separately and PKM2 protein expression was observed in whole cell extract, cytoplasmic extract and cytosolic extract after TMZ treatment. Relative expression of PKM2, relative expression of cytosolic phosphorylated histone H2AX (p⁃H2AX) and relative expression of siRNA knockdown PKM2 gene in U87MG and U251MG cells stably expressing the F3 ⁃ T3 fusion gene, detected by Western blotting. Results 1) CCK⁃8 cell proliferation assay showed that the survival rate of U87MG cells in the F3 ⁃ T3 transfected group was higher than that in the empty vector transfected group after treatment with TMZ 640, 320, 160, 80, 40 μmol/L (P = 0.000, 0.000, 0.000, 0.000, 0.004, 0.010), and the survival rate of U251MG cells in the F3 ⁃ T3 transfected group was also higher than that in the empty vector transfected group after TMZ 640, 320, 160, 80, 40, 20, 5 μmol/L (P = 0.000, 0.000, 0.000, 0.002, 0.001, 0.002); the survial rate of U87MG cells in the si⁃PKM2⁃1009 transfected group was lower than that of F3 ⁃ T3 transfected group after TMZ 640, 320, 160, 80, 40, 20, 10, 5, 2.50 μmol/L, the survival rate of U251MG cells in the si⁃PKM2⁃1377 transfected group was also lower than that in F3⁃T3 transfected group after TMZ 640, 320, 160, 80, 40, 20, 5 μmol/L (P = 0.000, 0.000, 0.002, 0.000, 0.002, 0.048, 0.042); and the survival rate of U87MG cells in TMZ + Compound 3k group was lower than TMZ group after TMZ 640, 320, 160, 80, 40, 20 μmol/L (P = 0.000, 0.000, 0.000, 0.000, 0.001, 0.002), and the survival rate of U251MG cells in TMZ + Compound 3k group after treatment with high concentrations of TMZ (640, 320, 160, 80 and 40 μmol/L) was also lower than TMZ group (P = 0.000, 0.000, 0.000, 0.000, 0.003), while the survival rate of U251MG cells in TMZ + Compound 3k group after treatment with low concentrations of TMZ (10, 5 and 2.50 μmol/L) was higher than that of TMZ group (P = 0.000, 0.000, 0.006). 2) An animal model of glioblastoma showed the presence of TMZ resistance in homozygous mice. 3) Bioinformatic analysis showed that the biological function of F3⁃T3 was significantly enriched in the DNA repair pathway (P = 0.000). Survival and overall survival of glioblastoma patients in the TCGA database were lower in the PKM2 high expression group than in the low expression group (P < 0.05). 4) Western blotting showed that the relative expression of p⁃H2AX in U87MG (P = 0.000, 0.000, 0.004) and U251MG (P = 0.000, 0.007, 0.005) cells in the F3 ⁃ T3 transfected group were lower than those in the empty vector transfected group after TMZ treatment for 48 h and then medium change for 24, 36 and 48 h. PKM2 protein incorporation into the nucleus was observed in both U87MG and U251MG cells in the F3⁃T3 transfected group after TMZ treatment, whereas it was not observed in cells in the empty vector transfected group; si⁃PKM2⁃1009 and si⁃ PKM2⁃1377 knocked down the relative expression of p⁃H2AX in U87MG (P = 0.000, 0.001, 0.006) and U251MG (P = 0.000, 0.000, 0.000) cells, respectively. Conclusions In the presence of TMZ, F3 ⁃ T3 promotes PKM2's nuclear translocation and activates DNA damage repair pathways, which in the eventually results resistance of glioblastoma cell to TMZ. PKM2 inhibitors can compromise the resistance glioblastoma cells stably expressing F3⁃T3 fusion gene to TMZ.

Abstract The devastating wheat stripe (yellow) rust pathogen, Puccinia striiformis f. sp. tritici (Pst), is a macrocyclic and heteroecious fungus. Pst produces urediniospores and teliospores on its primary host, wheat, and pycniospores and aeciospores are produced on its alternate hosts, barberry (Berberis spp.) or mahonia (Mahonia spp.). Basidiospores are developed from teliospores and infect alternate hosts. These five spore forms play distinct roles in Pst infection, disease development, and fungal survival, etc. However, the specific genes and mechanisms underlying these functional differences are largely unknown. In this study, we performed, for the first time in rust fungi, the deep RNA sequencing to examine the transcriptomic shift among all five Pst spore forms. Among a total of 29,591 identified transcripts, 951 were specifically expressed in basidiospores, whereas 920, 761, 266, and 110 were specific for teliospores, pycniospores, aeciospores, and urediniospores, respectively. Additionally, transcriptomes of sexual spores, namely pycniospores and basidiospores, showed significant differences from those of asexual spores (urediniospores, teliospores, and aeciospores), and transcriptomes of urediniospores and aeciospores were more similar to each other than to the three other spore forms. Especially, the basidiospores and pycniospores which infected the berberis shows wide differences in the cell wall degrading-enzymes and mating and pheromone response genes. Besides, we also found that there are 6234 differential expressed genes between the urediniospores and pycniospores, while only have 3 genes have alternative splicing enents, suggesting that differential genes expression may make more contribution than AS. This comprehensive transcriptome profiling can substantially improve our understanding of the developmental biology of the wheat stripe rust fungus.

Abstract Molecular diagnostics for crop diseases can guide the precise application of pesticides, thereby reducing pesticide usage while improving crop yield, but tools are lacking. Here, we report an in-field molecular diagnostic tool that uses a cheap colorimetric paper and a smartphone, allowing multiplexed, low-cost, rapid detection of crop pathogens. Rapid nucleic acid amplification-free detection of pathogenic RNA is achieved by combining toehold-mediated strand displacement with a metal ion-mediated urease catalysis reaction. We demonstrate multiplexed detection of six wheat pathogenic fungi and an early detection of wheat stripe rust. When coupled with a microneedle for rapid nucleic acid extraction and a smartphone app for results analysis, the sample-to-result test can be completed in ~10 min in the field. Importantly, by detecting fungal RNA and mutations, the approach allows to distinguish viable and dead pathogens and to sensitively identify mutation-carrying fungicide-resistant isolates, providing fundamental information for precision crop disease management.

Deep learning technology is widely used in the field of number recognition.It constructs neural network model through deep learning technology,nonlinear transformation activation function in neurons,different activation functions with different parameter initialization strategies,trains MINIST handwritten data set,constructs analysis model and recognizes numbers in images,reduce the dimension of a large amount of data into a small amount of data,and ensure the effective retention of image features.Through the analysis of image data,adding the feature conversion process,using the gradient descent optimizer to build a network structure and reduce the dimension of data,which can effectively avoid over fitting.Cross-entropy verification is used to compile and train the model,and the output classification results are further analyzed.Through the K-nearest neighbor classification algorithm,KNN classifier is set to further improve the accuracy of classification and prediction.Through MNIST data set experiment,the recognition rate is about 96.2%.The K-nearest neighbor algorithm(KNN) is introduced into the output layer,combined with the full connection layer and softmax layer of traditional convolutional neural network(CNN).After cross verification,the recognition rate is 99.6%.

Objective To investigate the effects of JSH⁃23 combined with Stattic targeting inhibition of nuclear factor⁃κB (NF⁃κB) and signal transducer and activator of transcription factor 3 (STAT3) signaling pathways on the proliferation and migration ability of mesenchymal glioblastoma cells and the expressions of NF⁃κB pathway and STAT3 pathway⁃related proteins. Methods The mRNA⁃seq results of 529 glioma patients were downloaded from The Cancer Genome Atlas (TCGA). Bioinformatics was used to analyze the correlation between RelA/P65 and STAT3 and the markers of mesenchymal glioblastoma, as well as the expression of NF⁃κB pathway and STAT3 pathway⁃related proteins in mesenchymal, classical and proneural glioblastoma. The human glioma cell lines U87MG and U251MG in vitro were treated with JSH⁃23, Stattic, JSH⁃23 and Stattic, respectively. The half⁃inhibitory concentration (IC50) of the two drugs was calculated by cytotoxicity assay, and the synergistic effect of the two drugs was observed by drug synergistic assay. CCK⁃8 assay and colony⁃forming assay were used to detect the proliferation activity of tumor cells. Wound healing assay and Transwell assay were used to detect the migration ability of tumor cells. Western blotting was used to detect the relative expressions of NF⁃κB pathway and STAT3 pathway⁃related proteins P65, phosphorylated P65 (p⁃P65), STAT3, phosphorylated STAT3 (p⁃STAT3) and CD44. Results 1) Bioinformatics analysis showed that RelA/P65 mRNA and STAT3 mRNA were positively correlated with mesenchymal glioblastoma markers CD44, CXCR4, CHI3L1, IL⁃4R and TRADD (r=0.206-0.605; P＜0.01, for all); the expressions of NF⁃κB pathway and STAT3 pathway⁃related proteins in mesenchymal glioblastoma were the highest, followed by classical glioblastoma, and the lowest in proneural glioblastoma. 2) The cytotoxicity assay showed the IC50 of JSH⁃23 on U87MG and U251MG cells were 59.39 and 56.21 μmol/L, and Stattic were 0.96 and 1.08 μmol/L. The drug synergistic assay showed that the synergistic effect of JSH⁃23 at 40-80 μmol/L and Stattic at 0.50-1 μmol/L on U87MG was the highest, and the synergistic effect of JSH⁃23 at 40 μmol/L and Stattic at 1 μmol/L on U251MG was the highest. The final concentration of JSH⁃23 was 60 μmol/L and Stattic was 1 μmol/L. 3) After the combined treatment of JSH⁃23, Stattic, JSH⁃23 and Stattic, the proliferation activity of U87MG and U251MG cells decreased (P=0.000, for all), the colony forming efficiency decreased (P=0.000, for all), the cell mobility decreased (P＜0.05, for all), and the positive cell numbers of crystal violet staining decreased (P=0.000, for all). The relative expressions of P65 (P=0.000, for all), p⁃P65 (P=0.000, for all), STAT3 (P=0.000, for all), p⁃STAT3 (P=0.000, for all) and CD44 (P=0.000, for all) were decreased, especially after JSH⁃23 combined with Stattic treatment (P＜0.01, for all). Conclusions JSH⁃23 combined with Stattic targeting inhibition of NF⁃κB and STAT3 signaling pathways can effectively inhibit the proliferation and migration ability of mesenchymal glioblastoma cells, and down⁃regulate the relative expressions of NF⁃κB pathway and STAT3 pathway⁃related proteins.

The antibiotic resistance of the food-borne pathogen Vibrio parahaemolyticus has attracted researchers’ attention in recent years, but its molecular mechanism remains poorly understood. In this study, 7 genes encoding outer membrane proteins (OMPs) were individually deleted in V. parahaemolyticus ATCC33846, and the resistance of these 7 mutants to 14 antibiotics was investigated. The results revealed that the resistance of the 7 mutants to ampicillin was significantly increased. Further exploration of 20-gene transcription changes by real time-qPCR (RT-qPCR) demonstrated that the higher ampicillin resistance might be attributed to the expression of β-lactamase and reduced peptidoglycan (PG) synthesis activity through reduced transcription of penicillin-binding proteins (PBPs), increased transcription of L,D-transpeptidases, downregulated D,D-carboxypeptidase, and alanine deficiency. This study provides a new perspective on ampicillin resistance in OMP mutants with respect to PG synthesis.

Electricity theft causes significant damage to social and economic development.How to detect malicious electricity theft based on power big data has been widely concerned by academia and industry.Aiming at the problems of traditional methods relying on manual features,insufficient behavior sequence representation,poor detection accuracy,etc.,this paper proposes an electricity theft detection model based on multi-head attention mechanism (ETD-MHA).The bidirectional gated recurrent unit is used to fully capture the time features of the electricity consumption behavior sequence,and the distinction of key features is gradually enhanced in the multi-head attention mechanism,and finally,the learning effect is improved by deepening the networks.Extended experiments are conducted on the smart meter datasets of Ireland and China State Grid.The results show that the proposed method achieves better performance compared with the linear regression (LR),support vector machine (SVM),random forest (RF),and other traditional algorithms.For example,the AUC value of the proposed model is improved by up to 34.6%compared to the LR algorithm.

In order to solve the problems encountered in the passage of thin-bedrock shallow coal seam roadways, three roadways in the 4-2 coal seam of Zhangjiamao Coal Mine were taken as an example, through drilling engineering, water pressure test, core identification, drilling video, hydrochemical test and other exploration techniques, the structure of overlying rock, weathering degree of bedrock, thickness of fresh bedrock and hydro-geological characteristics of weathered zone of 4-2 coal in roadway trench driving section were found out. Based on the exploration results, the technical ideas of “advanced grouting on the ground, repairing and remediation of surface river channels, and supplementary grouting in underground” were proposed, which avoided the hydro-geological and engineering geological problems encountered in the direct excavation of the roadway, which not only ensured the efficient and safe production of the mine, but also protected surface water resources.