Development and validation of polyamines metabolism-associated gene signatures to predict prognosis and immunotherapy response in lung adenocarcinoma.

Background: Polyamines metabolism is closely related to tumor development and progression, as well as tumor microenvironment (TME). In this study, we focused on exploring whether polyamines metabolism-associated genes would provide prognosis and immunotherapy response prediction in lung adenocarcinoma (LUAD). Methods: The expression profile data of polyamines metabolism-associated genes were acquired from the Cancer Genome Atlas (TCGA) database. Utilizing the least absolute shrinkage and selection operator (LASSO) algorithm, we created a risk score model according to polyamines metabolism-associated gene signatures. Meanwhile, an independent cohort (GSE72094) was employed to validate this model. Through the univariate and multivariate Cox regression analyses, the independent prognostic factors were identified. Subsequently, quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect their expression in LUAD cells. By consensus clustering analysis, polyamines metabolism-associated subgroups were determined in LUAD patients, with differential gene expression, prognosis, and immune characteristics analyses explored. Results: A total of 59 polyamines metabolism genes were collected for this study, of which 14 genes were identified for the construction of risk score model using LASSO method. High- and low- risk groups of LUAD patients in TCGA cohort were distinguished via this model, and high-risk group presented dismal clinical outcomes. The same prognostic prediction of this model had been also validated in GSE72094 cohort. Meanwhile, three independent prognostic factors (PSMC6, SMOX, SMS) were determined for constructing the nomogram, and they were all upregulated in LUAD cells. In addition, two distinct subgroups (C1 and C2) were identified in LUAD patients. Comparing the two subgroups, 291 differentially expressed genes (DEGs) were acquired, mainly enriching in organelle fission, nuclear division, and cell cycle. Comparing to C1 subgroup, the patients in C2 subgroup had favorable clinical outcomes, increased immune cells infiltration, and effective immunotherapy response. Conclusion: This study identified polyamines metabolism-associated gene signatures for predicting the patients' survival, and they were also linked to immune cells infiltration and immunotherapy response in LUAD patients.

Experimental study on the suppressive effect of B3GNT3 on the apoptosis of lung adenocarcinoma cells and its application in early screening for lung adenocarcinoma.

Background: Lung adenocarcinoma, as a common histological type in lung cancer, the overall survival is very low, and the prognosis is poor because it is difficult to find and easily recurs. Therefore, this study aimed to explore the role of the secreted protein beta-1,3-N-acetylglucosaminyltransferase 3 (B3GNT3) in the development of lung adenocarcinoma and to evaluate its potential significance for early clinical biomarker screening. Methods: The mRNA expression profiles of patients with lung adenocarcinoma and normal controls were analyzed via The Cancer Genome Atlas (TCGA) database. Serum samples of clinical lung cancer patients and healthy people were obtained, and the differences in B3GNT3 expression in different stages of lung adenocarcinoma and in healthy tissues were compared. Kaplan-Meier (K-M) curves were drawn to clarify the influence of high and low expression of B3GNT3 on the prognosis of patients. Peripheral blood samples from patients with lung adenocarcinoma and healthy people were obtained clinically, and receiver operating characteristic (ROC) curves were drawn to clarify the sensitivity and specificity of B3GNT3 expression for the diagnosis of lung adenocarcinoma. Lung adenocarcinoma cells were cultured in vitro, the expression of B3GNT3 was knocked down by lentivirus infection. The expression of the apoptosis-associated genes was detected by reverse transcription-polymerase chain reaction (RT-PCR). Results: The secreted protein B3GNT3 is significantly differentially expressed in the serum of patients with lung adenocarcinoma versus normal controls. Subgroup analysis according to lung adenocarcinoma clinical stage showed that the higher the clinical stage of lung adenocarcinoma was, the higher the B3GNT3 expression. Enzyme-linked immunosorbent assay (ELISA) revealed that B3GNT3 expression was significantly increased in the serum of patients with lung adenocarcinoma and significantly decreased after surgery. By inhibiting programmed cell death-ligand 1 (PD-L1), the level of apoptosis was significantly increased and the proliferative capacity was significantly inhibited. In contrast, the level of apoptosis was significantly increased and the proliferation ability was significantly inhibited after simultaneous overexpression of B3GNT3 and inhibition of PD-L1. Conclusions: High expression of the secreted protein B3GNT3 in lung adenocarcinoma is closely related to prognosis and can serve as a potential biological marker for the early screening of lung adenocarcinoma.

Characterization of Fatigue Crack Growth Based on Acoustic Emission Multi-Parameter Analysis.

In engineering structures that are subject to cyclic loading, monitoring and assessing fatigue crack growth (FCG) plays a crucial role in ensuring reliability. In this study, the acoustic emission (AE) technique was used to monitor the FCG behavior of 2.25Cr1Mo0.25V steel in real-time. Specifically, an AE multi-parameter analysis was conducted to qualitatively assess the crack growth condition and quantitatively correlate the crack growth rate with AE. Various AE parameters were extracted from AE signals, and the performances of different AE parameters were analyzed and discussed. The results demonstrated that four stages of FCG, which correspond to macrocrack initiation, stable crack growth with low crack growth rate, stable crack growth with high crack growth rate, and unstable crack growth, are distinctly identified by several AE time domain parameters. The sudden and continuous occurrence of many AE signals with high count (>100) and high energy (>40 mV·ms) can provide early and effective warning signs for accelerated crack growth before final failure occurs. Moreover, linear correlations between crack growth rate and different AE parameters are established for quantifying crack growth. Based on the AE multi-parameter analysis, it was found that the count, energy, and kurtosis are superior AE parameters for both qualitatively and quantitatively characterizing the FCG in 2.25Cr1Mo0.25V steel. Results from this research provide an AE strategy based on multi-parameter analysis for effective monitoring and assessment of FCG in engineering materials.

Mesonephric-Like Adenocarcinoma of Uterine Corpus: A Clinicopathological and Targeted Genomic Profiling Study in a Single Institution.

Background: Mesonephric-like adenocarcinoma (MLA) is a recently characterized, rare, and aggressive neoplasm that mostly arises in the uterine corpus and ovary. MLA shows characteristic pathological features similar to mesonephric adenocarcinoma of the cervix. The origin of MLA is still controversial and recognition of it remains challenging for pathologists. The aim of this study was to enrich the clinicopathological features of MLA in the uterine corpus and explore its molecular alterations by targeted next-generation sequencing (NGS). Methods: Four cases of MLA were identified among a total of 398 endometrial carcinomas diagnosed in our institution between January 2014 and December 2021. Immunohistochemistry and targeted NGS spanning 437 cancer-relevant genes were performed. Results: The most common symptom was abnormal vaginal bleeding, and the average age was 68 years. Histologically, the tumors showed a mixture of varied growth patterns including papillary, glandular, tubular, cribriform, solid, and slit-like architectures, which were lined by columnar to cuboidal cells with overlapping vesicular nuclei and sometimes nuclear grooves. Intraluminal eosinophilic colloid-like secretions were focally evident in three of the four cases. Immunohistochemically, the MLAs were positive for GATA3 (4/4), TTF-1 (3/3), luminal CD10 (3/3), calretinin (2/3), and patchy P16 (3/3) and were negative for ER (0/4) and PR (0/4). The expression of P53 was "wild type" (4/4). By targeted NGS, 3/4 (75%), 2/4 (50%), and 1/4 (25%) cases harbored PIK3CA, KRAS, and PTEN mutations, respectively. None of the tumors had mutations in DNA mismatch repair genes, ARID1A/B, POLE, CTNNB1, SMARCA4, or TP53. At the time of diagnosis, three were presented with FIGO IB stage and one with IIIC stage. Two patients received postoperative chemotherapy and radiotherapy and they were alive without evidence of disease at 8 and 56 months follow-up, respectively. One patient developed pulmonary metastasis 13 months after surgery and chemotherapy, and one was dead of the disease 24 months after the operation without adjuvant therapy. Conclusions: MLA is a rare and aggressive malignancy, representing approximately 1% of all endometrial carcinomas. It exhibits mixed architectures associated with distinctive immunophenotype and recurrent KRAS and PIK3CA mutations, supporting classified as of Müllerian origin with mesonephric differentiation.

FGF21 alleviates pulmonary hypertension by inhibiting mTORC1/EIF4EBP1 pathway via H19.

Long non-coding RNAs (lncRNAs) play a significant role in pulmonary hypertension (PH). Our preliminary data showed that hypoxia-induced PH is attenuated by fibroblast growth factor 21 (FGF21) administration. Therefore, we further investigated the regulatory role of long non-coding RNAs in PH treated with FGF21. RNA sequencing analysis and real-time PCR identified a significantly up-regulation of the H19 after FGF21 administration. Moreover, gain- and loss-of-function assays demonstrated that FGF21 suppressed hypoxia-induced proliferation of pulmonary artery smooth muscle cells partially through upregulation of H19. In addition, FGF21 deficiency markedly exacerbated hypoxia-induced increases of pulmonary artery pressure and pulmonary vascular remodelling. In addition, AAV-mediated H19 overexpression reversed the malignant phenotype of FGF21 knockout mice under hypoxia expose. Further investigation uncovered that H19 also acted as an orchestra conductor that inhibited the function of mechanistic target of rapamycin complex 1 (mTORC1) by disrupting the interaction of mTORC1 with eukaryotic translation initiation factor 4E-binding protein 1 (EIF4EBP1). Our work highlights the important role of H19 in PH treated with FGF21 and suggests a mechanism involving mTORC1/EIF4EBP1 inhibition, which may provide a fundamental for clinical application of FGF21 in PH.

Swin-UNet++: A Nested Swin Transformer Architecture for Location Identification and Morphology Segmentation of Dimples on 2.25Cr1Mo0.25V Fractured Surface.

The precise identification of micro-features on 2.25Cr1Mo0.25V steel is of great significance for understanding the mechanism of hydrogen embrittlement (HE) and evaluating the alloy's properties of HE resistance. Presently, the convolution neural network (CNN) of deep learning is widely applied in the micro-features identification of alloy. However, with the development of the transformer in image recognition, the transformer-based neural network performs better on the learning of global and long-range semantic information than CNN and achieves higher prediction accuracy. In this work, a new transformer-based neural network model Swin-UNet++ was proposed. Specifically, the architecture of the decoder was redesigned to more precisely detect and identify the micro-feature with complex morphology (i.e., dimples) of 2.25Cr1Mo0.25V steel fracture surface. Swin-UNet++ and other segmentation models performed state-of-the-art (SOTA) were compared on the dimple dataset constructed in this work, which consists of 830 dimple scanning electron microscopy (SEM) images on 2.25Cr1Mo0.25V steel fracture surface. The segmentation results show Swin-UNet++ not only realizes the accurate identification of dimples but displays a much higher prediction accuracy and stronger robustness than Swin-Unet and UNet. Moreover, efforts from this work will also provide an important reference value to the identification of other micro-features with complex morphologies.

A Nomogram Integrating Ferroptosis- and Immune-Related Biomarkers for Prediction of Overall Survival in Lung Adenocarcinoma.

Ferroptosis plays a dual role in cancer, which is known to be affected to antitumor immune responses. However, the association between ferroptosis and antitumor immune responses is uncertain in lung adenocarcinoma (LUAD). In this work, 38 ferroptosis-related genes (FRGs) and 429 immune-related genes (IRGs) were identified as being differentially expressed between tumor and normal samples. Two risk score formulas consisting of seven FRGs and four IRGs, respectively, were developed by Lasso-penalized Cox regression and verified in the GSE13213 dataset. The CIBERSORT algorithm was used to estimate the relative abundance of immune cells in tumors. The correlation between FRGs and immune cells was evaluated using the TIMER database. The results indicated that the development of ferroptosis was synergistic with that of anti-tumor immunity in LUAD. The concordance index and calibration curves showed that the performance of a nomogram that combines clinical staging and risk scores is superior to that of models using a single prognostic factor. In conclusion, ferroptosis might be synergistic with anti-tumor immunity in LUAD. The combined nomogram could reliably predict the probability of overall survival of LUAD patients. These findings may be useful for future investigation of prognostic value and therapeutic potential related to ferroptosis and tumor immunity in LUAD.

Experimental Investigation of Fatigue Crack Growth Behavior of the 2.25Cr1Mo0.25V Steel Welded Joint Used in Hydrogenation Reactors.

In this work, the fatigue crack growth (FCG) behavior and fatigue damage mechanism of the 2.25Cr1Mo0.25V steel welded joint used in hydrogenation reactors were investigated. The multi-pass welding was carried out to manufacture the welded joint using the combined shielded metal arc welding (SMAW) and submerged automatic arc welding (SAAW) processes. The FCG behavior of different zones in the welded joint, including the base metal (BM), the heat-affected zone (HAZ) and the weld metal (WM), were studied by compact tension tests. Moreover, the acoustic emission (AE) technique was used to monitor AE signals generated from FCG process for further understanding FCG behavior and fatigue mechanisms. Additionally, the microstructures and fracture surfaces of different specimens were observed by optical microscopy (OM) and scanning electron microscopy (SEM). The results revealed that the microstructure of BM is fine granular bainite, while the WM shows coarser bainite grains. The HAZ exhibits the most significant inhomogeneity with large dispersion of grain size. FCG results showed that the HAZ exhibits much higher fatigue crack growth rate (FCGR) at low ΔK values, while the BM shows the most superior fatigue resistance. The AE technique is successful in monitoring and identifying damage evolutions during the FCG process. Moreover, an enhanced AE activity is observed in FCG of the WM specimen, which is attributed to the combined influence of the formation of numerous secondary cracks and coarse-grained microstructures.

High-Temperature Tensile and Creep Behavior in a CrMoV Steel and Weld Metal.

The 2.25Cr1Mo0.25V steel is a vanadium-modified 2.25Cr1Mo steel and is being widely used in the manufacture of heavy-wall hydrogenation reactors in petrochemical plants. However, the harsh service environment requires a thorough understanding of high-temperature tensile and creep behaviors of 2.25Cr1Mo0.25V steel and its weld for ensuring the safety and reliability of hydrogenation reactors. In this work, the high-temperature tensile and creep behaviors of base metal (BM) and weld metal (WM) in a 2.25Cr1Mo0.25V steel weldment used for a hydrogenation reactor were studied experimentally, paying special attention to its service temperature range of 350-500 °C. The uniaxial tensile tests under different temperatures show that the WM has higher strength and lower ductility than those of BM, due to the finer grain size in the WM. At the same time, the short-term creep tests at 550 °C reveal that the WM has a higher creep resistance than that of BM. Moreover, the creep damage mechanisms were clarified by observing the fracture surface and microstructures of crept specimens with the aid of scanning electron microscopy (SEM). The results showed that the creep damage mechanisms of both BM and WM are the initiation and growth of creep cavities at the second phase particles. Results from this work indicate that the mismatch in the high-temperature tensile strength, ductility, and creep deformation rate in 2.25Cr1Mo0.25V steel weldment needs to be considered for the design and integrity assessment of hydrogenation reactors.

Screening and verifying key genes with poor prognosis in colon cancer through bioinformatics analysis.

BACKGROUND: Colon cancer (CC) is one of the tumors with high morbidity and mortality in the world, and has a trend of younger generation. The molecular level of CC has not been fully elaborated. The purpose of this study is to screen and identify important genes with poor prognosis and their mechanisms at different levels. METHODS: GSE74602 and GSE10972 gene expression profiles were downloaded from the Gene Expression Omnibus (GEO) database. There were 58 normal tissues and 58 CC tissues. Differentially expressed genes (DEGs) were screened out by using the GEO2R tool and Venn diagram. Then, the DAVID online database was used to perform the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Six hub genes with the highest correlation were screened out after the modular analysis of the protein-protein interaction (PPI) network by using Cytoscape's MCODE plug-in. Finally, the overall survival of key hub genes and potential pathways were verified in GEPIA and UALCAN database. RESULTS: A total of 78 up-regulated DEGs were enriched in the mitotic nuclear division, cell division, cell proliferation, anaphase-promoting complex-dependent catabolic process and G2/M transition of the mitotic cell cycle. In total, 130 down-regulated DEGs were enriched in muscle contraction, bicarbonate transport, cellular response to zinc ion, negative regulation of growth, negative regulation of leukocyte apoptotic process and one-carbon metabolic process. CDK1, CCNB1, CDC20, AURKA, CCNA2 and TOP2A were the top six hub genes, mainly enriched in cell cycle pathways. Among them, CCNB1, CDK1, CDC20, CCNA2 were enriched in the G2/M phase. GEPIA and UALCAN database confirmed that CCNA2 and CCNB1 had a significant relationship with the poor prognosis of CC patients. Meanwhile, there was a positive correlation between the two. CONCLUSIONS: Screening out genes with abnormal expression in CC help understand the initiation and progression of CC at the molecular level and explore candidate biomarkers for diagnosis, treatment and prognosis.

Investigation of the Influence of Pre-Charged Hydrogen on Fracture Toughness of As-Received 2.25Cr1Mo0.25V Steel and Weld.

Fracture failure caused by hydrogen embrittlement (HE) is a major concern for the system reliability and safety of hydrogen storage vessels, which are generally made of 2.25Cr1Mo0.25V steel. Thus, study of the influence of pre-charged hydrogen on fracture toughness of as-received 2.25Cr1Mo0.25V steel and weld is of significant importance. In the current work, the influence of hydrogen on fracture toughness of as-received 2.25Cr1Mo0.25V steel and weld was systematically studied. Base metal (BM) and weld metal (WM) specimens under both hydrogen-free and hydrogen-charged conditions were tested using three-point bending tests. Hydrogen was pre-charged inside specimens by the immersion charging method. The J-integral values were calculated for quantitatively evaluating the fracture toughness. In order to investigate the HE mechanisms, optical microscopy (OM) and scanning electron microscopy (SEM) were used to characterize the microstructure of BM and WM specimens. The results revealed that the presence of pre-charged hydrogen caused a significant decrease of the fracture toughness for both BM and WM specimens. Moreover, the pre-charged hydrogen led to a remarkable transition of fracture mode from ductile to brittle pattern in 2.25Cr1Mo0.25V steel.

Effect of Cementite on the Hydrogen Diffusion/Trap Characteristics of 2.25Cr-1Mo-0.25V Steel with and without Annealing.

Hydrogen embrittlement (HE) is a critical issue that affects the reliability of hydrogenation reactors. The hydrogen diffusivity/trap characteristics of 2.25Cr-1Mo-0.25V steel are important parameters mainly used to study the HE mechanism of steel alloys. In this work, the hydrogen diffusivity/trap characteristics of heat-treated (annealed) and untreated 2.25Cr-1Mo-0.25V steel were studied using an electrochemical permeation method. The microstructures of both 2.25Cr-1Mo-0.25V steels were investigated by metallurgical microscopy. The effect of cementite on the hydrogen diffusivity/trap mechanisms was studied using thermodynamics-based and Lennard⁻Jones potential theories. The results revealed that the cementite located at the grain boundaries and at the interfaces of lath ferrite served as a kind of hydrogen trap (i.e., an irreversible hydrogen trap). In addition, hydrogen was transported from ferrite to cementite via up-hill diffusion, thereby supporting the hypothesis of cementite acting as a hydrogen trap.

Experimental Investigation of the Effect of Hydrogen on Fracture Toughness of 2.25Cr-1Mo-0.25V Steel and Welds after Annealing.

Hydrogen embrittlement (HE) is a critical issue that hinders the reliability of hydrogenation reactors. Hence, it is of great significance to investigate the effect of hydrogen on fracture toughness of 2.25Cr-1Mo-0.25V steel and weld. In this work, the fracture behavior of 2.25Cr-1Mo-0.25V steel and welds was studied by three-point bending tests under hydrogen-free and hydrogen-charged conditions. The immersion charging method was employed to pre-charge hydrogen inside specimen and the fracture toughness of these joints was evaluated quantitatively. The microstructure and grain size of the specimens were observed by scanning electron microscopy (SEM) and by metallurgical microscopy to investigate the HE mechanisms. It was found that fracture toughness for both the base metal (BM) and the weld zone (WZ) significantly decreased under hydrogen-charged conditions due to the coexistence of the hydrogen-enhanced decohesion (HEDE) and hydrogen-enhanced localized plasticity (HELP) mechanisms. Moreover, the formation and growth of primary voids were observed in the BM, leading to a superior fracture toughness. In addition, the BM compared to the WZ shows superior resistance to HE because the finer grain size in the BM leads to a larger grain boundary area, thus distributing more of the diffusive hydrogen trapped in the grain boundary and reducing the hydrogen content.