Detection of BRAF Mutations on Direct Smears of Thyroid Fine Needle Aspirates through Cell Transfer Technique

Objectives: To determine the utility of the cell transfer technique (CTT) for BRAF molecular testing on thyroid fine-needle aspiration (FNA) specimens. Methods: Polymerase chain reaction (PCR)–based BRAF molecular testing was performed on tissues obtained through CTT from both air-dried and ethanol-fixed direct smears of thyroid FNA specimens and then compared with the corresponding thyroidectomy formalin-fixed, paraffin-embedded (FFPE) tissues on 30 cases. Results: BRAF testing was successfully performed on 29 of 30 air-dried CTT, 27 of 30 ethanol-fixed CTT, and 27 of 30 FFPE tissues. The results exhibited 11, 13, and 13 BRAF mutations and 18, 14, and 14 wild types for the air-dried CTT, the ethanol-fixed CTT, and the FFPE tissues, respectively. The concordance rate was 96% between air-dried and ethanol-fixed CTT tissues, 88% between air-dried CTT and FFPE tissues, and 92% between ethanol-fixed CTT and FFPE tissues. Conclusions: PCR-based BRAF mutational testing can be reliably performed on the direct smears of the thyroid FNA specimens through the application of CTT

Dub3 Inhibition Suppresses Breast Cancer Invasion and Metastasis by Promoting Snail1 Degradation

Snail1, a key transcription factor of epithelial–mesenchymal transition (EMT), is subjected to ubiquitination and degradation, but the mechanism by which Snail1 is stabilized in tumours remains unclear. We identify Dub3 as a bona fide Snail1 deubiquitinase, which interacts with and stabilizes Snail1. Dub3 is overexpressed in breast cancer; knockdown of Dub3 resulted in Snail1 destabilization, suppressed EMT and decreased tumour cell migration, invasion, and metastasis. These effects are rescued by ectopic Snail1 expression. IL-6 also stabilizes Snail1 by inducing Dub3 expression, the specific inhibitor WP1130 binds to Dub3 and inhibits the Dub3-mediating Snail1 stabilization in vitroand in vivo. Our study reveals a critical Dub3–Snail1 signalling axis in EMT and metastasis, and provides an effective therapeutic approach against breast cancer

Identification and Characterization of Two Novel Compounds: Heterozygous Variants of Lipoprotein Lipase in Two Pedigrees With Type I Hyperlipoproteinemia

BackgroundType I hyperlipoproteinemia, characterized by severe hypertriglyceridemia, is caused mainly by loss-of-function mutation of the lipoprotein lipase (LPL) gene. To date, more than 200 mutations in the LPL gene have been reported, while only a limited number of mutations have been evaluated for pathogenesis.ObjectiveThis study aims to explore the molecular mechanisms underlying lipoprotein lipase deficiency in two pedigrees with type 1 hyperlipoproteinemia.MethodsWe conducted a systematic clinical and genetic analysis of two pedigrees with type 1 hyperlipoproteinemia. Postheparin plasma of all the members was used for the LPL activity analysis. In vitro studies were performed in HEK-293T cells that were transiently transfected with wild-type or variant LPL plasmids. Furthermore, the production and activity of LPL were analyzed in cell lysates or culture medium.ResultsProband 1 developed acute pancreatitis in youth, and her serum triglycerides (TGs) continued to be at an ultrahigh level, despite the application of various lipid-lowering drugs. Proband 2 was diagnosed with type 1 hyperlipoproteinemia at 9 months of age, and his serum TG levels were mildly elevated with treatment. Two novel compound heterozygous variants of LPL (c.3G>C, p. M1? and c.835_836delCT, p. L279Vfs*3, c.188C>T, p. Ser63Phe and c.662T>C, p. Ile221Thr) were identified in the two probands. The postheparin LPL activity of probands 1 and 2 showed decreases of 72.22 ± 9.46% (p<0.01) and 54.60 ± 9.03% (p<0.01), respectively, compared with the control. In vitro studies showed a substantial reduction in the expression or enzyme activity of LPL in the LPL variants.ConclusionsTwo novel compound heterozygous variants of LPL induced defects in the expression and function of LPL and caused type I hyperlipoproteinemia. The functional characterization of these variants was in keeping with the postulated LPL mutant activity

Toward the development of smart capabilities for understanding seafloor stretching morphology and biogeographic patterns via DenseNet from high-resolution multibeam bathymetric surveys for underwater vehicles

The increasing use of underwater vehicles facilitates deep-sea exploration at a wide range of depths and spatial scales. In this paper, we make an initial attempt to develop online computing strategies to identify seafloor categories and predict biogeographic patterns with a deep learning-based architecture, DenseNet, integrated with joint morphological cues, with the expectation of potentially developing its embedded smart capacities. We utilized high-resolution multibeam bathymetric measurements derived from MBES and denoted a collection of joint morphological cues to help with semantic mapping and localization. We systematically strengthened dominant feature propagation and promoted feature reuse via DenseNet by applying the channel attention module and spatial pyramid pooling. From our experiment results, the seafloor classification accuracy reached up to 89.87%, 82.01%, and 73.52% on average in terms of PA, MPA, and MIoU metrics, achieving comparable performances with the state-of-the-art deep learning frameworks. We made a preliminary study on potential biogeographic distribution statistics, which allowed us to delicately distinguish the functionality of probable submarine benthic habitats. This study demonstrates the premise of using underwater vehicles through unbiased means or pre-programmed path planning to quantify and estimate seafloor categories and the exhibited fine-scale biogeographic patterns

Association of the Genomic Profile of Medullary Thyroid Carcinoma with Tumor Characteristics and Clinical Outcomes in an International Multicenter Study

Purpose: The prognostic importance of RET and RAS mutations and their relationship to clinicopathologic parameters and outcomes in medullary thyroid carcinoma (MTC) need to be clarified. Experimental Design: A multicenter retrospective cohort study was performed utilizing data from 290 patients with MTC. The molecular profile was determined and associations were examined with clinicopathologic data and outcomes. Results: RET germ line mutations were detected in 40 patients (16.3%). Somatic RET and RAS mutations occurred in 135 (46.9%) and 57 (19.8%) patients, respectively. RETM918T was the most common somatic RET mutation (n = 75). RET somatic mutations were associated with male sex, larger tumor size, advanced American Joint Committee Cancer (AJCC) stage, vascular invasion, and high International Medullary Thyroid Carcinoma Grading System (IMTCGS) grade. When compared with other RET somatic mutations, RETM918T was associated with younger age, AJCC (eighth edition) IV, vascular invasion, extrathyroidal extension, and positive margins. RET somatic or germ line mutations were significantly associated with reduced distant metastasis-free survival on univariate analysis, but there were no significant independent associations on multivariable analysis, after adjusting for tumor grade and stage. There were no significant differences in outcomes between RET somatic and RET germ line mutations, or between RETM918T and other RET mutations. Other recurrent molecular alterations included TP53 (4.2%), ARID2 (2.9%), SETD2 (2.9%), KMT2A (2.9%), and KMT2C (2.9%). Among them, TP53 mutations were associated with decreased overall survival (OS) and disease-specific survival (DSS), independently of tumor grade and AJCC stage. Conclusions: RET somatic mutations were associated with high-grade, aggressive primary tumor characteristics, and decreased distant metastatic-free survival but this relationship was not significant after accounting for tumor grade and disease stage. RETM918T was associated with aggressive primary tumors but was not independently associated with clinical outcomes. TP53 mutation may represent an adverse molecular event associated with decreased OS and DSS in MTC, but its prognostic value needs to be confirmed in future studies

Diagnostic Value of MAML2 Rearrangements in Mucoepidermoid Carcinoma

Mucoepidermoid carcinoma (MEC) is often seen in salivary glands and can harbor MAML2 translocations (MAML2+). The translocation status has diagnostic utility as an objective confirmation of the MEC diagnosis, for example, when distinction from the more aggressive adenosquamous carcinoma (ASC) is not straightforward. To assess the diagnostic relevance of MAML2, we examined our 5-year experience in prospective testing of 8106 solid tumors using RNA-seq panel testing in combinations with a two-round Delphi-based scenario survey. The prevalence of MAML2+ across all tumors was 0.28% (n = 23/8106) and the majority of MAML2+ cases were found in head and neck tumors (78.3%), where the overall prevalence was 5.9% (n = 18/307). The sensitivity of MAML2 for MEC was 60% and most cases (80%) were submitted for diagnostic confirmation; in 24% of cases, the MAML2 results changed the working diagnosis. An independent survey of 15 experts showed relative importance indexes of 0.8 and 0.65 for “confirmatory MAML2 testing” in suspected MEC and ASC, respectively. Real-world evidence confirmed that the added value of MAML2 is a composite of an imperfect confirmation test for MEC and a highly specific exclusion tool for the diagnosis of ASC. Real-world evidence can help move a rare molecular-genetic biomarker from an emerging tool to the clinic

PcmSU-A Packet Classification Method Supporting High-Speed Search and Fast Update

Packet classification is the core technology that underpins software-defined networking (SDN). With the rapid development of network applications, the increasing complexity of flow tables in SDN brings challenges to update and classification time. In order to achieve fast search and update at the same time, this article proposes a high-performance packet classification method PcmSU based on decision tree. The method optimizes the search and update process in the traditional decision tree-based classification method, and the classification efficiency of the constructed decision tree has been greatly improved compared with that before optimization. Based on the characteristics of the small interval, the reconstruction of the decision tree is greatly reduced and the rule update efficiency is improved. In addition, based on the characteristics of the decision tree constructed by this method, there is no need to consider the specific location and sequence number of rule updates when adding rules, which can avoid introducing rule conflicts. Experimental results show that the PcmSU method not only supports high-speed packet classification and linear memory consumption, but also has fast rule update speed

A review of environment problems in the coastal sea of South China

The coastal sea of South China provided an important habitat for protection and propagation of marine organisms. Rapid economic development and human activities, such as wastewater discharge, reclamation, overfishing and aquaculture in the South China Sea had already resulted in environmental degradation, and thus caused sharp contradictions between exploitation and protection of the coastal sea of South China. In this present article, the main environment problems and degradation trends were reviewed based on literatures and other sources of information, which mainly referenced nutrient pollution, persistent organic pollution and metal pollution, decrease of biodiversity, reduction of marine habitat and frequent natural and ecological disasters. The current efforts in China on protecting the environment in the coastal sea of South China were discussed, which included improving legislation by formulating a series of laws and regulations at national or local level, setting up natural reserves, and supporting research projects. There were many challenges regarding policy, management and science research to protect and sustain the coastal sea of South China, such as imperfect legal and administrative systems, lack of public participation, poor financial support and lack of monitoring and evaluation. Finally, some recommendations were put forward for the sake of the sustainable use of the environment in the coastal sea of South China, including reinforcing the planning of marine resource exploitation and use through integrated coastal zone management, strengthening the marine environment and protection awareness of the public, and scientifically establishing the fishery spawning spots and aquatic reserves.The coastal sea of South China provided an important habitat for protection and propagation of marine organisms. Rapid economic development and human activities, such as wastewater discharge, reclamation, overfishing and aquaculture in the South China Sea had already resulted in environmental degradation, and thus caused sharp contradictions between exploitation and protection of the coastal sea of South China. In this present article, the main environment problems and degradation trends were reviewed based on literatures and other sources of information, which mainly referenced nutrient pollution, persistent organic pollution and metal pollution, decrease of biodiversity, reduction of marine habitat and frequent natural and ecological disasters. The current efforts in China on protecting the environment in the coastal sea of South China were discussed, which included improving legislation by formulating a series of laws and regulations at national or local level, setting up natural reserves, and supporting research projects. There were many challenges regarding policy, management and science research to protect and sustain the coastal sea of South China, such as imperfect legal and administrative systems, lack of public participation, poor financial support and lack of monitoring and evaluation. Finally, some recommendations were put forward for the sake of the sustainable use of the environment in the coastal sea of South China, including reinforcing the planning of marine resource exploitation and use through integrated coastal zone management, strengthening the marine environment and protection awareness of the public, and scientifically establishing the fishery spawning spots and aquatic reserves

EGF/EGFR upregulates and cooperates with Netrin-4 to protect glioblastoma cells from DNA damage-induced senescence

BackgroundGlioblastoma multiforme (GBM) is the most malignant central nervous system tumor. Alkylating agent, temozolomide (TMZ), is currently the first-line chemotherapeutic agent for GBM. However, the sensitivity of GBM cells to TMZ is affected by many factors. And, several clinic trials, including co-administration of TMZ with other drugs, have failed in successful treatment of GBM. We have previously reported that Netrin-4 (NTN4), a laminin-like axon guidance protein, plays a protective role in GBM cell senescence upon TMZ-triggered DNA damage. However, the master regulator of NTN4 needs further elucidation. Epidermal growth factor/Epidermal growth factor receptor (EGF/EGFR) can modulate the expression of various extracellular matrix related molecules, and prevent DNA damage in GBM cells. In this study, we investigated the relationship between EGF/EGFR signaling and NTN4, and explored their effect on therapeutic efficacy in GBM cells upon TMZ treatment.MethodsCo-expression analysis were performed by using the RNA sequencing data from NIH 934 cell lines and from single cell RNA sequencing data of GBM tumor. The co-expressing genes were used for GO enrichment and signaling pathway enrichment. mRNA expression of the target genes were quantified by qPCR, and cell senescence were investigated by Senescence-Associated Beta-Galactosidase Staining. Protein phosphorylation were observed and analyzed by immunoblotting. The RNA sequencing data and clinical information of TMZ treated patients were extracted from TCGA-glioblastoma project, and then used for Kaplan-Meier survival analysis.ResultsAnalysis of RNA sequencing data revealed a potential co-expression relationship between NTN4 and EGFR. GO enrichment of EGFR-correlated genes indicated that EGFR regulates GBM cells in a manner similar to that in central nervous system development and neural cell differentiation. Pathway analysis suggested that EGFR and its related genes contribute to cell adhesion, extracellular matrix (ECM) organization and caspase related signaling. We also show that EGF stimulates NTN4 expression in GBM cells and cooperates with NTN4 to attenuate GBM cell senescence induced by DNA damage, possibly via AKT and ERK. Clinical analysis showed that co-expression of EGFR and NTN4 significantly predicts poor survival in TMZ-treated GBM patients.ConclusionsThis study indicates that EGF/EGFR regulates and cooperates with NTN4 in DNA damage resistance in GBM. Therefore, our findings provide a potential therapeutic target for GBM.Peer reviewe

Sediment Carbon Sequestration and Driving Factors in Seagrass Beds from Hainan Island and the Xisha Islands

Seagrass beds are considered to be substantial sinks of “blue carbon”. However, differentiation in the carbon sink capacities of seagrass beds in different regions with distinct nutrient conditions remains unclear. In this study, sediment carbon stocks, seagrass biomass, and microbial community structures and potential functions of seagrass beds in eutrophic seawater adjacent to Hainan Island and oligotrophic seawater around the Xisha Islands were compared. Our results showed that sediment mineralizable organic carbon and dry bulk density were substantially higher on Hainan Island than on the Xisha Islands (t-test, p p > 0.05). Similarly, seagrass biomass was much higher on Hainan Island (p p p < 0.05). Proteobacteria, Desulfobacterota, Ascomycota and Basidiomycota could account for degrading organic carbon on Hainan Island. Proteobacteria and Bacteroidota may contribute primarily to carbon loss in the seagrass beds of the Xisha Islands. This study strengthens our understanding of the effects of human activities on carbon sequestration in seagrass bed ecosystems