Circulating EGFR Mutations in Patients with Lung Adenocarcinoma by Circulating Tumor Cell Isolation Systems: A Concordance Study.

Background: We developed a hybrid platform using a negative combined with a positive selection strategy to capture circulating tumor cells (CTCs) and detect epidermal growth factor receptor (EGFR) mutations in patients with metastatic lung adenocarcinoma. Methods: Blood samples were collected from patients with pathology-proven treatment-naïve stage IV lung adenocarcinoma. Genomic DNA was extracted from CTCs collected for EGFR mutational tests. The second set of CTC-EGFR mutational tests were performed after three months of anti-cancer therapy. Results: A total of 80 samples collected from 28 patients enrolled between July 2016 and August 2018. Seventeen patients had EGFR mutations, including Exon 19 deletion (n = 11), L858R (n = 5), and de-novo T790 and L858R (n = 1). Concordance between tissue and CTCs before treatment was 88.2% in EGFR- mutant patients and 90.9% in non-mutant patients. The accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of EGFR mutation tests for CTCs were 89.3%, 88.2%, 90.9%, 93.8%, and 83.3%, respectively. Conclusions: CTCs captured by a hybrid platform using a negative and positive selection strategy may serve as a suitable and reliable source of lung cancer tumor DNA for detecting EGFR mutations, including T790M.

A safety comparative study between freezing nitrogen ethanol composite and liquid nitrogen for cryotherapy of musculoskeletal tumors.

Freezing nitrogen ethanol composite (FNEC) showed effective cryoablative ability for bone tumor ex vivo and in vivo comparable to liquid nitrogen (LN). We therefore wished to compare the radiant cooling damage of the surrounding tissue between FNEC and LN. The evaluation of the radiant cooling damage was demonstrated human bone xenograft transplantation (HXT) in a mouse model. Characterizations and quantifications of the damaging effects on morphologic features and apoptosis of the cryoablative surrounding bone tissue, muscle and epidermal layer of skin were compared. The radiant cooled damaging effects including epidermal rupture, hair follicle atrophy, dermis and subcutaneous crystal vacuolation of skin were significantly greater in LN than FNEC. Muscular apoptosis, structural shrinkage and bone cellular apoptosis were supposedly 15%-33% destroying degrees of LN more than FNEC. We concluded that FNEC is an innovative cryogenic material, and it could cause less cryoablative damage to surrounding normal tissue than LN. The findings might support the safety of FNEC being applied in clinical cryoablation therapy.

Distinct effects of Disabled-2 on transferrin uptake in different cell types and culture conditions.

Iron uptake by the transferrin (Tf)-transferrin receptor (TfR) complex is critical for erythroid differentiation. The mechanisms of TfR trafficking have been examined, but the adaptor proteins involved in this process are not fully elucidated. We have investigated the role of the adaptor protein, Disabled-2 (Dab2), in erythroid differentiation and Tf uptake in the cells of hematopoietic lineage. Dab2 was upregulated in a time-dependent manner during erythroid differentiation of mouse embryonic stem cells and human K562 erythroleukemic cells. Attenuating Dab2 expression in K562 cells diminished TfR internalization and increased surface levels of TfR concomitantly with a decrease in Tf uptake and erythroid differentiation. Dab2 regulated Tf uptake of the suspended, but not adherent, cultures of K562 cells. In contrast, Dab2 is not involved in TfR trafficking in the HeLa cells with epithelial origin. These differential effects are Dab2-specific because attenuating the expression of adaptor protein 2 µ subunit inhibited the uptake of Tf regardless of culture condition. We offer novel insight of Dab2 function in iron uptake and TfR internalization for the suspended culture of hematopoietic lineage cells.

The Prognostic Value of Circulating Tumor Cells in Asian Neuroendocrine Tumors.

Circulating tumor cells (CTC) play important roles in various cancers; however, few studies have assessed their clinical utility in neuroendocrine tumors. This study aimed to prospectively evaluate the prognostic value of CTC counts in Asian patients with neuroendocrine tumors before and during anti-cancer therapy. Patients who were diagnosed with unresectable histological neuroendocrine tumors between September 2011 and September 2017 were enrolled. CTC testing was performed before and during anti-cancer therapy using a negative selection protocol. Chromogranin A levels were also assessed. Univariate and multivariate Cox's proportional hazard model with forward LR model was performed to investigate the impact of independent factors on overall survival and progression-free survival. Kaplan-Meier method with log-rank tests were used to determine the difference among different clinicopathological signatures and CTC cutoff. The baseline CTC detection rate was 94.3% (33/35). CTC counts were associated with cancer stages (I-III vs. IV, P = 0.015), liver metastasis (P = 0.026), and neuroendocrine tumor grading (P = 0.03). The median progression-free survival and overall survivals were 12.3 and 30.4 months, respectively. In multivariate Cox regression model, neuroendocrine tumors grading and baseline CTC counts were both independent prognostic factors for progression-free survival (PFS, P = 0.005 and 0.015, respectively) and overall survival (OS, P = 0.018 and 0.023, respectively). In Kaplan-Meier analysis, lower baseline chromogranin A levels were associated with longer PFS (P = 0.024). Baseline CTC counts are associated with the clinicopathologic features of neuroendocrine tumors and are an independent prognostic factor for this malignancy.

A Delphi-based approach to developing expert systems with the cooperation of multiple experts.

Knowledge acquisition has been a critical bottleneck in building knowledge-based systems. In past decades, several methods and systems have been proposed to cope with this problem. Most of these methods and systems were proposed to deal with the acquisition of domain knowledge from single expert. However, as multiple experts may have different experiences and knowledge on the same application domain, it is necessary to elicit and integrate knowledge from multiple experts in building an effective expert system. Moreover, the recent literature has depicted that "time" is an important parameter that might significantly affect the accuracy of inference results of an expert system; therefore, while discussing the elicitation of domain expertise from multiple experts, it becomes an challenging and important issue to take the "time" factor into consideration. To cope with these problems, in this study, we propose a Delphi-based approach to eliciting knowledge from multiple experts. An application on the diagnosis of Severe Acute Respiratory Syndrome has depicted the superiority of the novel approach.

A Mobile Sleep-Management Learning System for Improving Students' Sleeping Habits by Integrating a Self-Regulated Learning Strategy: Randomized Controlled Trial.

BACKGROUND: Insomnia can significantly affect students' learning performance. Researchers have indicated the importance and challenge of coping with insomnia using nondrug treatments, such as cognitive behavioral therapy (CBT) for insomnia. However, it is easy for the traditional CBT for insomnia to be interrupted owing to the overly lengthy period of sleep therapy. Self-regulated learning (SRL) strategies are known to be an effective approach for helping students improve their time management, as well as their ability to set learning goals and adopt learning strategies. OBJECTIVE: The objective of this study was to propose a mobile sleep-management learning system integrated with SRL strategies and CBT. METHODS: A total of 18 undergraduate students from a university in northern Taiwan participated in the 2-week experiment of using this sleep-management system. RESULTS: The experimental results showed that the proposed approach was useful and easy for students to use. In addition, the number of students with insomnia significantly decreased; that is, the proposed approach could help students improve their sleep quality and cultivate better sleeping habits, which is important for them to enhance their learning efficiency. CONCLUSIONS: With the assistance of this proposed approach, students can plan their daily life by setting goals, applying strategies, monitoring their life habits process, and modifying strategies to cultivate good learning and healthy lifestyle habits. TRIAL REGISTRATION: Government Research Bulletin MOST104-3011-E038-001; https://www.grb.gov.tw/search/planDetail? id=11568383&docId=467988 (Archived by WebCite at http://www.webcitation.org/73MnPHNri).

Erythroid differentiation is augmented in Reelin-deficient K562 cells and homozygous reeler mice.

Reelin is an extracellular glycoprotein that is highly conserved in mammals. In addition to its expression in the nervous system, Reelin is present in erythroid cells but its function there is unknown. We report in this study that Reelin is up-regulated during erythroid differentiation of human erythroleukemic K562 cells and is expressed in the erythroid progenitors of murine bone marrow. Reelin deficiency promotes erythroid differentiation of K562 cells and augments erythroid production in murine bone marrow. In accordance with these findings, Reelin deficiency attenuates AKT phosphorylation of the Ter119(+)CD71(+) erythroid progenitors and alters the cell number and frequency of the progenitors at different erythroid differentiation stages. A regulatory role of Reelin in erythroid differentiation is thus defined.