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Objective To analyze the correlation of interferon-induced protein with tetratricopeptide repeats 1 (IFIT1) with immune infiltration and prognosis of ovarian cancer (OC). Methods GEO database was employed to select the tumor immune related genes, and Kaplan-Meier and Prognoscan databases were used to identify the genes significantly associated with OC prognosis. The differential expression of IFIT1 between OC tissue and normal tissue were confirmed with GEPIA, Human Protein Atlas, and Timer databases. The expression level of IFIT1 in OC tissues with different grades and stages were analyzed in the UALCAN database. In addition, based on David database, GO enrichment analysis was used to analyze the interacting genes and proteins of IFIT1 in the String and Genemania databases. Timer and Tisidb databases verified the correlation between IFIT1 and immune cells mutually. Finally, after IFIT1 knockdown xenograft model was constructed based on lentiviral vector of IFIT1 shRNA, the tumor growth was observed in the transplanted nude mice, and infiltration of neutrophils was observed with immunohistochemical assay. Results FIT1, a tumor immune gene, selected from the GEO database, Kaplan-Meier and Prognoscan databases, was negatively correlated with the OC prognosis. GEPIA, Human Protein Atlas, Timer database, and UALCAN database indicated that the expression level of IFIT1 was significantly higher in the OC tissues than the normal ovarian tissues, and had no obvious correlation with tumor stage and grade. Analysis in String, Genemania, and David database found the interaction genes and proteins of IFIT1 were enriched in activation of 2'-5' oligonucleotide synthase, virus defense, and innate immunity, and other processes. The Timer database presented that IFIT1 was positively correlated with the infiltration of CD8+T cells, B cells, dendritic cells, neutrophils, and macrophage in OC, with neutrophils having the most significant correlation. Tisidb and GSCA also confirmed the positive correlation between IFIT1 and neutrophil infiltration in OC (P < 0.05). RT-qPCR and Western blotting confirmed that IFIT1 knockdown resulted in suppressed tumor growth in OC cells xenograft nude mice (P < 0.05) and reduced neutrophil infiltration in the xenograft tissues. Conclusion IFIT1 may influence the malignant progression of OC by promoting neutrophil infiltration.

Background: Irradiation disrupts the vascular niche where hematopoietic stem cells (HSCs) reside, causing delayed hematopoietic reconstruction. The subsequent recovery of sinusoidal vessels is key to vascular niche regeneration and a prerequisite for hematopoietic reconstruction. We hypothesize that resident bone marrow macrophages (BM-Mφs) are responsible for repairing the HSC niche upon irradiation injury. Methods: We examined the survival and activation of BM-Mφs in C57BL/6 mice upon total body irradiation. After BM-Mφ depletion via injected clodronate-containing liposomes and irradiation injury, hematopoietic reconstruction and sinusoidal vascular regeneration were assessed with immunofluorescence and flow cytometry. Then enzyme-linked immunosorbent assay (ELISA) and flow cytometry were performed to analyze the contribution of VEGF-A released by BM-Mφs to the vascular restructuring of the HSC niche. VEGF-A-mediated signal transduction was assessed with transcriptome sequencing, flow cytometry, and pharmacology (agonists and antagonists) to determine the molecular mechanisms of Piezo1-mediated responses to structural changes in the HSC niche. Results: The depletion of BM-Mφs aggravated the post-irradiation injury, delaying the recovery of sinusoidal endothelial cells and HSCs. A fraction of the BM-Mφ population persisted after irradiation, with residual BM-Mφ exhibiting an activated M2-like phenotype. The expression of VEGF-A, which is essential for sinusoidal regeneration, was upregulated in BM-Mφs post-irradiation, especially CD206+ BM-Mφs. The expression of mechanosensory ion channel Piezo1, a response to mechanical environmental changes induced by bone marrow ablation, was upregulated in BM-Mφs, especially CD206+ BM-Mφs. Piezo1 upregulation was mediated by the effects of irradiation, the activation of Piezo1 itself, and the M2-like polarization induced by the phagocytosis of apoptotic cells. Piezo1 activation was associated with increased expression of VEGF-A and increased accumulation of NFATC1, NFATC2, and HIF-1α. The Piezo1-mediated upregulation in VEGF-A was suppressed by inhibiting the calcineurin/NFAT/HIF-1α signaling pathway. Conclusion: These findings reveal that BM-Mφs play a critical role in promoting vascular niche regeneration by sensing and responding to structural changes after irradiation injury, offering a potential target for therapeutic efforts to enhance hematopoietic reconstruction.

PurposeRepairing the irradiation-induced osteogenic differentiation injury of bone marrow mesenchymal stem cells (BM-MSCs) is beneficial to recovering haematopoiesis injury in radiotherapy; however, its mechanism is elusive. Our study aimed to help meet the needs of understanding the effects of radiotherapy on BM-MSC osteogenic potential.Methods and materialsBalb/c mice and the BM-MSCs were used to evaluate the irradiation-induced osteogenic differentiation injury in vivo. The cellular and molecular characterization were applied to determine the mechanism for recovery of irradiation-derived haematopoiesis injuries.ResultsWe report a functional role of IL-12 in acute irradiation hematopoietic injury recovery and intend to dissect the possible mechanisms through BM-MSC, other than the direct effect of IL-12 on hematopoietic stem and progenitor cells (HSPCs). Specifically, we show that early use of IL-12 enhanced the osteogenic differentiation of BM-MSCs through IL-12Rβ1/TYK2/STAT3 signaling; furthermore, IL-12 induced osteogenesis facilitated bone formation and irradiation hematopoiesis recovery when transplanted BM-MSCs in the femur of Balb/c mice. For the mechanism of action, we found that IL-12 receptor beta 1 (IL-12Rβ1) expression of irradiated BM-MSCs was upregulated rapidly, coincidentally consistent with early use of IL-12 induced osteogenic differentiation enhancement. IL-12Rβ1 and tyrosine kinase 2 gene (Tyk2) silencing experiments and phosphotyrosine of signal transducer and activator of transcription 3 (p-STAT3) suppression experiments indicated the IL-12Rβ1/TYK2/STAT3 signaling was essential in IL-12-induced osteogenic differentiation enhancement of BM-MSCs.ConclusionThese findings suggested that IL-12 may exert BM-MSCs-based hematopoietic recovery by repairing osteogenic differentiation abilities damages through IL-12Rβ1/TYK2/STAT3 signaling pathway post-irradiation.

Objective To investigate the biological and metabolic roles of WNK2 in regulation of ovarian cancer based on transcriptome sequencing technology. Methods Ovarian cancer SKOV3 cells were divided into experimental group and control group, with 3 replicates in each group.The expression of WNK2 was knocked down with transfection of siRNA targeting WNK2 in SKOV3 cells.Then the total RNA was extracted and prepared for transcriptome sequencing and bioinformatics analysis after 48 h of transfection.The differential genes between siNC and siWNK2 were screened out in the range of padj < 0.01, fold change >1, and FPKM >500 to obtain the genes under the regulation of WNK2.DisGeNET and disease ontology (DO) enrichment analysis was used to analyze the relationship of the diseases and differentially expressed genes (DEGs).GO enrichment analysis was adopted to analyze the cellular functions and metabolic processes in which the DEGs were enriched.Finally, RT-qPCR was used to examine whether the 10 differential proteins in the reactive oxygen species (ROS) pathway were regulated by WNK2.The influence of WNK2 overexpression and knockdown on the ROS metabolic process of ovarian cancer cells was determined. Results The DisGeNET and DO enrichment analysis showed the obtained DEGs were closely related with malignant tumors (P < 0.05).GO functional enrichment analysis showed the obtained DEGs were enriched in biological processes such as ROS metabolism, apoptosis signaling pathway, formation of ubiquitin ligase complex and mitochondrial membrane protein complex, growth factor binding, and transcription activator activity and so on (P < 0.05).Among these enriched pathways, ROS related pathways were the most numerous and significant (P < 0.001).The results of RT-qPCR indicated that the changes in 9 DEGs were consistent with the sequencing results after WNK2 was knocked down, and one gene (PDGFB) was contrary to the sequencing results, and one gene had no significant change between the 2 groups.Knockdown of WNK2 in SKOV3 and CAOV3 cells resulted in decreased production of ROS, whereas overexpression of WNK2 increased the production. Conclusion WNK2 probably promotes ovarian cancer malignancy by inducing ROS metabolism.

Resistance to ionizing radiation (IR), which is a conventional treatment for osteosarcoma that cannot be resected, undermines the efficacy of this therapy. However, the mechanism by which IR induces radioresistance in osteosarcoma is not defined. Here, we report that CR6-interacting factor-1 (CRIF1) is highly expressed in osteosarcoma and undergoes nuclear-cytoplasmic shuttling of cyclin-dependent kinase 2 (CDK2) after IR. Osteosarcoma cells lacking CRIF1 show increased sensitivity to IR, which is associated with delayed DNA damage repair, inactivated G1/S checkpoint, and mitochondrial dysfunction. CRIF1 interacts with the DNA damage checkpoint regulator CDK2, and CRIF1 and CDK2 colocalize in the nucleus after IR. Nuclear localization of CDK2 is associated with phosphorylation changes that promote DNA repair and activation of the G1/S checkpoint. CRIF1 knockdown synergized with IR in an in vivo osteosarcoma model, leading to tumor regression. Based on these findings, we identify CRIF1 as a potential therapeutic target in osteosarcoma that can increase the efficacy of radiotherapy. More broadly, our findings may provide insights into the mechanism for other types of radioresistant cancers and be exploited for therapeutic ends.

Resistance to ionizing radiation (IR), which is a conventional treatment for osteosarcoma that cannot be resected, undermines the efficacy of this therapy. However, the mechanism by which IR induces radioresistance in osteosarcoma is not defined. Here, we report that CR6-interacting factor-1 (CRIF1) is highly expressed in osteosarcoma and undergoes nuclear-cytoplasmic shuttling of cyclin-dependent kinase 2 (CDK2) after IR. Osteosarcoma cells lacking CRIF1 show increased sensitivity to IR, which is associated with delayed DNA damage repair, inactivated G1/S checkpoint, and mitochondrial dysfunction. CRIF1 interacts with the DNA damage checkpoint regulator CDK2, and CRIF1 and CDK2 colocalize in the nucleus after IR. Nuclear localization of CDK2 is associated with phosphorylation changes that promote DNA repair and activation of the G1/S checkpoint. CRIF1 knockdown synergized with IR in an in vivo osteosarcoma model, leading to tumor regression. Based on these findings, we identify CRIF1 as a potential therapeutic target in osteosarcoma that can increase the efficacy of radiotherapy. More broadly, our findings may provide insights into the mechanism for other types of radioresistant cancers and be exploited for therapeutic ends.

Abstract Radiation induces rapid bone loss and enhances bone resorption and RANKL expression. RANKL provides the crucial signal to induce osteoclast differentiation and plays an important role in bone resorption. However, the mechanisms of radiation-induced osteoporosis are not fully understood. Here, we show that Crif1 expression increases in bone marrow cells after radiation. Conditional Crif1 deletion in bone marrow cells causes decreases in RANKL expression and the RANKL/OPG ratio, and relieves bone loss after radiation in mice. We further demonstrated in vitro that Crif1 promotes RANKL secretion via the cAMP/PKA pathway. Moreover, protein-protein docking screening identified five compounds as Crif1 inhibitors; these compounds dramatically suppressed RANKL secretion and CREB phosphorylation when cells were exposed to forskolin. This study enriches current knowledge of the pathogenesis of osteoporosis and provides insights into potential therapeutic strategies for osteoporosis treatment.

Objective To analyze the biological processes regulated by protein kinase WNK2 in ovarian cancer cells based on protein phosphorylation modified mass spectrometry. Methods Ovarian cancer cell line SKOV3 was transfected with lentiviral vector to knockdown WNK2 and the obtained cells were further screened by drug screening. Then the total protein was extracted for protein phosphorylation modification mass spectrometry analysis. Bioinformatics analysis was adopted to detect the phosphorylation changes before and after WNK2 knockdown, and differential proteins were screened out. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were employed to explore the functions of differentially phosphorylated proteins, and experiments were performed to verify the biological processes regulated by WNK2. Results Phosphorylation modified mass spectrometry indicated that WNK2 knockdown significantly decreased the phosphorylation of 745 proteins with 1 078 altered protein sites, and increased the phosphorylation of 328 proteins with 415 altered protein sites. KEGG and GO enrichment analyses showed WNK2 influences biological processes such as cytoskeleton organization, signal transduction, cell composition, and regulation of kinase activity. Phalloidin staining indicated after WNK2 knockdown resulted in broken and disordered cytoskeleton, while its overexpression promoted cytoskeleton remodeling. Conclusion WNK2 is involved in the cytoskeleton formation in ovarian cancer cells by regulating the phosphorylation of vimentin and other proteins.

In this paper, the structure design and optimization of the column of the tree-shaped parking garage are carried out. The base dimensions are 2.4 m、0.7 m and 8.5 m respectively. The simplified model of column takes into account the axial stress caused by the total weight FN of each component and the bending stress caused by the suspended loading plate and the vehicle. The maximum stress is easy to occur at the root of the column. The calculated results are 63.06 MPa, less than allowable stress 117.5 MPa, maximum deflection 8.93 mm, much less than standard value 113.90 mm. In order to meet the requirements of strength and deflection displacement, the penalty function method is used to optimize the size of the main support of the column, and the optimized model is measured and compared with the software analysis. Under the same working condition, the stress change data at the root of the column are collected by the test instrument and compared with the software analysis. The stress at the same position is less than 10% of the deviation between the measured and the simulated analysis results. The design and analysis of column structure are carried out by combining finite element analysis with actual stress test, and the mixed penalty function method is used to optimize the column structure. The results show that the method is effective. It provides some means and reference for the analysis of this kind of project.

Simple Summary: Xinjiang Brown cattle are a unique and widely distributed breed of dual-purpose cattle in Xinjiang, the People's Republic of China. It is known that this cattle breed has different milk production performance compared to Holstein cattle. However, the microbiota differences between Xinjiang Brown and Holstein cattle are not well known. Therefore, this study aimed to compare the bacterial community composition of the rumen and feces of these two cattle breeds under the same dietary and management conditions using 16s rRNA sequencing. This study identified microbiota with different relative abundance between these two cattle breeds and their biological functions might be related to milk synthesis. Our study increases the understanding of the differences in bacterial microbiota between Xinjiang Brown and Holstein cattle. Xinjiang Brown cattle are a unique and widely distributed breed of dual-purpose cattle in the Xinjiang area of China, whose milk production performance differs from Holstein cattle. It has been known that variations in bacterial species of the gastrointestinal tract influence milk protein, fat, and lactose synthesis. However, the microbiota differences between Xinjiang Brown and Holstein cattle are less known. This study aims to compare the bacterial community composition of the rumen and feces of these two cattle breeds under the same dietary and management conditions. The 16s rRNA sequencing data and milk production of 18 Xinjiang Brown cows and 20 Holstein cows on the same farm were obtained for analysis. The results confirmed differences in milk production between Xinjiang Brown and Holstein cattle. Microbiota with different relative abundance between these two cattle breeds were identified, and their biological functions might be related to milk synthesis. This study increases the understanding of the differences in microbiota between Xinjiang Brown and Holstein cattle and might provide helpful information for microbiota composition optimization of these dairy cattle. [ABSTRACT FROM AUTHOR]

Application Programming Interface (API) migration is a common task for adapting software across different programming languages and platforms, where manually constructing the mapping relations between APIs is indeed time-consuming and error-prone. To facilitate this process, many automated API mapping approaches have been proposed. However, existing approaches were mainly designed and evaluated for mapping APIs of statically-typed languages, while their performance on dynamically-typed languages remains unexplored. In this article, we conduct the first extensive study to explore existing API mapping approaches' performance for mapping APIs in dynamically-typed languages, for which we have manually constructed a high-quality dataset. According to the empirical results, we have summarized several insights. In particular, the source code implementations of APIs can significantly improve the effectiveness of API mapping. However, due to the confidentiality policy, they may not be available in practice. To overcome this, we propose a novel API mapping approach, named Matl, which leverages the transfer learning technique to learn the semantic embeddings of source code implementations from large-scale open-source repositories and then transfers the learned model to facilitate the mapping of APIs. In this way, Matl can produce more accurate API embedding of its functionality for more effective mapping without knowing the source code of the APIs. To evaluate the performance of Matl, we have conducted an extensive study by comparing Matl with state-of-the-art approaches. The results demonstrate that Matl is indeed effective as it improves the state-of-the-art approach by at least 18.36% for mapping APIs of dynamically-typed language and by 30.77% for mapping APIs of the statically-typed language. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression of folate receptor 1(FOLR1) and mesothelin (MSLN) in epithelial ovarian cancer(EOC) and explore their clinical significance. Methods The data of clinical samples and transcriptome sequencing expression matrix of EOC were obtained from TCGA and GEO database(Ovarian Cancer TCGA, Provisional; GSE6008; GSE66957; GSE49997) to analyze the expressions of FOLR1 and MSLN in different tissues and their association with tumor staging and the prognosis of EOC. We also collected paraffin-embedded tissue samples of EOC(105 cases), ovarian borderline tumor(11 cases), and normal ovarian tissue(30 cases) from patients undergoing surgeries in our hospital between 2018 and 2019. The expression levels of FOLR1 and MSLN were detected by immunohistochemistry and their correlation with the clinicopathological characteristics of the patients was analyzed. Results Analysis of TCGA and GEO databases showed that the mRNA levels of FOLR1 and MSLN were significantly higher in EOC tissues than in normal ovarian tissues(P < 0.05), and was also higher in stage Ⅲ~Ⅳ than in stage Ⅰ~Ⅱ EOC(P < 0.05). Survival analysis showed that the patients with a high expression of FOLR1 had significantly longer overall survival(P < 0.05) and disease-free survival(P < 0.05), while the patients with a high expression of MSLN had significantly shortened overall survival(P < 0.05) and disease-free survival(P < 0.05). Immunohistochemical analysis showed that FOLR1 and MSLN proteins were lowly expressed in normal ovarian tissues, and their expressions increased moderately in borderline ovarian tumor and significantly in EOC tissues. The positive expression rates of FOLR1 and MSLN in EOC were 80.95% and 53.33%, respectively. The levels were of FOLR1 and MSLN differed significantly in different tissue types(P < 0.001), and their positive rates were significantly higher in stage Ⅲ~Ⅳ than in stage Ⅰ~Ⅱ EOC(P < 0.05). The positive rate of MSLN was significantly increased in patients with lymph node metastasis(P < 0.05). Subgroup survival analysis of TCGA database showed that patients with a low MSLN expression and a high FOLR1 expression(MLFH) at the mRNA level had the best prognosis, while those with a high MSLN expression and a low FOLR1 expression(MHFL) had the worst prognosis(median survival time 70.57 vs 25.85 months, P < 0.000 1); compared with the MLFL patients, the MHFL patients in the early stage(< 50 months) had a better prognosis, while beyond 50 months, the survival rate of the patients decreased, but the difference was not statistically significant. Conclusion In patients with EOC, a high expression of FOLR1 indicates a more favorable prognosis while a high expression of MSLN is associated with a poor prognosis. These 2 proteins may participate in the process of EOC, and their combined detection may help in prognostic evaluation of EOC.