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Zhaohua Xiao,1 Xumei Feng,2 Yongjia Zhou,1 Peiwei Li,3 Junwen Luo,1 Wenhao Zhang,1 Jie Zhou,1 Jiangfeng Zhao,1 Dong Wang,4 Yongjie Wang,5 Zhongxian Tian,1,6 Xiaogang Zhao1,6 1Department of Thoracic Surgery, The Second Hospital of Shandong University, Jinan, 250033, People’s Republic of China; 2Health Management Center, The Second Hospital of Shandong University, Jinan, People’s Republic of China; 3Institute of Medical Sciences, The Second Hospital of Shandong University, Jinan, People’s Republic of China; 4Department of Thoracic Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, 250021, People’s Republic of China; 5Department of Thoracic Surgery, The Affiliated Hospital of Qingdao University, Qingdao, 266000, People’s Republic of China; 6Key Laboratory of Chest Cancer, Shandong University, The Second Hospital of Shandong University, Jinan, People’s Republic of ChinaCorrespondence: Zhongxian Tian; Xiaogang Zhao, Department of Thoracic Surgery, The Second Hospital of Shandong University, Jinan, 250033, People’s Republic of China, Tel +86-17660082365 ; +86-053185875009, Email tianzhongxian@email.sdu.edu.cn; zhaoxiaogang@sdu.edu.cnBackground: Cancer cell-derived exosomal microRNAs (miRNAs) play critical role in orchestrating intercellular communication between tumor cells and tumor microenvironmental factors, including lymphatic endothelial cells (LECs). Nevertheless, the functions and underlying mechanisms of exosomal miRNAs in lymphatic metastasis and lymphangiogenesis in esophageal squamous cell carcinoma (ESCC) remain unclear.Methods: Small RNA sequencing, Gene Expression Omnibus (GEO) analysis and qRT‒PCR were performed to identify the candidate exosomal miRNAs involved in ESCC metastasis. Receiver operating characteristic curve analysis was conducted to evaluate the diagnostic potential of exosomal miR-10527-5p in predicting lymph node metastasis (LNM) status. An in vitro coculture system was used to investigate the effects of exosomal miR-10527-5p on ESCC cells and human LECs (HLECs), followed by a popliteal LNM assay in vivo. The relationship between miR-10527-5p and Rab10 was identified by dual-luciferase reporter, fluorescence in situ hybridization and qRT‒PCR assays. Then, a series of rescue assays were performed to further investigate whether Rab10 is involved in exosomal miR-10527-5p mediated ESCC metastasis.Results: MiR-10527-5p was found to be notably reduced in both the plasma exosomes and tumor tissues of ESCC patients with LNM, and plasma exosomal miR-10527-5p had a high sensitivity and specificity for discrimination of LNM status. Moreover, exosome-shuttled miR-10527-5p suppressed the migration, invasion and epithelial-to-mesenchymal transition (EMT) of ESCC cells as well as the migration and tube formation of HLECs via Wnt/β-catenin signaling in vitro and in vivo. Further investigation revealed that Rab10 was a direct target of miR-10527-5p, and re-expression of Rab10 neutralized the inhibitory effects of exosomal miR-10527-5p.Conclusion: Our study demonstrated that exosomal miR-10527-5p had a strong capability to predict preoperative LNM status and anti-lymphangiogenic effect. Exosomal miR-10527-5p inhibited lymphangiogenesis and lymphatic metastasis of ESCC in a vascular endothelial growth factor-C (VEGF-C)-independent manner, showing potential as a therapeutic target for ESCC patients.Keywords: esophageal squamous cell carcinoma, exosomes, miR-10527-5p, biomarker, lymphangiogenesis

BACKGROUND: The mutation rate of the tumor protein P53 (TP53) has been reported to be greater than 50% in non-small cell lung cancer (NSCLC), and gain-of-function (GOF) mutations in unfolded P53 (TP53(R175H) and TP53(Y220C)) have been associated with poor prognosis. However, the best treatment for patients with NSCLC harboring unfolded mutant P53 (mutp53) remains unclear. Triptolide is a natural compound derived from Tripterygium wilfordii that has shown a strong antitumor effect in a variety of cancers. Our study aimed to explore the GOF mutations in unfolded mutp53 (TP53(R175H) and TP53(Y220C)) and to clarify the molecular mechanisms by which triptolide regulates the degradation of unfolded mutp53 proteins in NSCLC. METHODS: Two unfolded proteins harboring TP53(R175H) and TP53(Y220C) mutations were selected to explore their functions in NSCLC progression. NCI-H1299 cells (TP53-null) were transfected with wild-type TP53 (TP53(WT)), TP53(R175H), or TP53(Y220C) genes and treated with triptolide or a vehicle. Wound healing and transwell assays were performed to measure cell migration and invasion in vitro. Lung metastasis models were constructed through tail vein injection of mutant cells into BALB/c nude mice to evaluate the effect of triptolide on metastasis in vivo. Western blotting, quantitative real-time polymerase chain reaction (qRT-PCR), immunoprecipitation, and dual-luciferase reporter assays were performed to explore the relevant molecular mechanisms. RESULTS: Our study revealed that triptolide treatment reduced TP53(R175H) levels and that the TP53(Y220C) mutation enhanced the invasion and migration of NCI-H1299 cells. Mechanistically, triptolide promoted TP53(R175H) and TP53(Y220C) protein proteasomal degradation mediated through the E3 ligase murine double minute 2 (MDM2) by directly interacting with heat shock protein 70 (HSP70). Moreover, by upregulating HSP70 transcription, triptolide contributed to the protein degradation of the GOF mutp53. CONCLUSIONS: Our study reports, for the first time, the mechanism underlying triptolide-regulated protein degradation of TP53(R175H) or TP53(Y220C), which offers new insight into developing a better therapeutic strategy for patients with NSCLC who express the unfolded mutp53 GOF protein.

Lung adenocarcinoma (LUAD) is the predominant pathological subtype of lung cancer, which is the most prevalent and lethal malignancy worldwide. Cyclins have been reported to regulate the physiology of various types of tumors by controlling cell cycle progression. However, the key roles and regulatory networks associated with the majority of the cyclin family members in LUAD remain unclear. In total, 556 differentially expressed genes were screened from the GSE33532, GSE40791 and GSE19188 mRNA microarray datasets by R software. Subsequently, protein-protein interaction network containing 499 nodes and 4,311 edges, in addition to a significant module containing 76 nodes and 2,631 edges, were extracted through the MCODE plug-in of Cytoscape. A total of four cyclin family genes [

Product design is an important part of the manufacturing system from a macro point of view. In the design process, creative design is one of the important factors to survive the fierce competition. During creative design process, designers are the most critical component. However, design fixation often negatively influences the design outcomes. Many researches from cognitive science and design science reveal that the presentation of outer information can alleviate design fixation effectively. Among different kinds of outer information, language terms are proved to be effective. This work presents a method of automatically generating language terms as design inspirations based on 500,000 granted patents. This method adopts evolutionary computation as the fundamental algorithm to retrieve language terms from a vocabulary base. To implement the algorithm, the vocabulary is first encoded by high dimensional vectors through word embedding, which is a three layers neural network. Further, two metrics for measuring the inspiration potential of language terms are defined in a computable manner. This work also conducts experiments to validate the method, and the experimental results show that (1) the algorithm is efficient and has the potential to be extended to larger vocabulary; (2) the generated design inspirations have a positive influence on the design outcomes.

Earlier studies have shown that the degree of abstract knowledge has a significant impact on the result of innovative design, which plays an important role in the process of design thinking, thereby affecting the generation of innovative design. This paper presents a design experiment that was conducted to study the effects of abstract knowledge to individual’s design thinking and the cognitive behavior. In the experiment, subjects were given different levels of abstract knowledge, respectively from three aspects of the product: Function-Behavior-Structure, and then they were asked to complete a design task. Then analyze the role of abstract knowledge’s form of cognition, and explore the mechanism of inspired design thinking further. The paper finally provides a theoretical basis for the design of innovative knowledge-oriented services.