Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Regional Coverage Planning and Obstacle Avoiding for Cleaning Robot in a Partially Known Environment

[[abstract]]本文研究居家清掃機器人在已知複雜居家環境之清掃規劃與行進控制，包括清掃區域規劃、行進路徑控制、機構設計、機電運動控制等課題。為提升清掃效率與導航控制，首先運用已知的環境參數建立電子地圖，藉由房門與通道特徵指定行進控制節點，以規劃出多個清掃區域與其清掃順序。在室內導航部分，則運用區域節點進行路徑規畫，配合臨時障礙的避障策略，以快速找到前往指定位置的最佳路徑。為克服室內設計常見的高架地板與門檻障礙，本研究開發具有攀爬機構設計的機器人為載具，在行進控制中能自動偵測可攀爬的障礙，利用跨越機構的展開與收合，跨越高架地板或門檻。機器人利用單晶片PIC16F877為基本動作控制核心，透過碰撞感測器與六個紅外線感測器判斷障礙物類型與位置，完成直行、轉向、循邊與跨越的動作。再藉由遠端伺服電腦上MATLAB的GUI 環境，設計整合與控制介面，以藍芽通訊，利用馬達的編碼器來估計機器人的方位，再將規劃路徑以模組化指令結構進行導航控制。最後透過製作原型實體與機電整合控制，來測試清掃機器人執行運動行為的能力，與驗證本控制架構的可行性。[[abstract]]This paper addresses regional coverage planning and obstacle avoiding for domestic cleaning robot in a partially known environment, including cleaning regional planning, path planning, mechanism design, motion control, and mechatronic control. To increase efficiency of cleaning and navigation, we first set up an electronic map from given environmental parameters. Nodal points are designated in doors and corridors for connected rooms for regional clean sequencing. Path planning among regional nodes and a strategy for provisional obstacle avoidance are proposed for fast indoor navigation. In order to negotiate staged floor and doorsill common in oriental domestic layouts, this thesis develops a cleaning robot with striding mechanism. By unfolding and folding of the striding arms, the robot can negotiate multiple rooms with level difference. The robot uses a single-chip PIC16F877 as a basic core for motion control. Contact sensors and six infrared sensors are arranged in the robot to detect types and locations of obstacles. A control module in a remote server PC is designed using MATLAB GUI environment, and communicates with the robot through Bluetooth technology. The encoder information of two actuators is used to estimate the location and orientation of the robot. The control module decodes the navigation path into parameterized structural commands. Finally, this study presents a prototype robot and the navigation system to demonstrate the feasibility and the functionality of the proposed scheme

Promoting dentinogenesis of DPSCs through inhibiting microRNA-218 by using magnetic nanocarrier delivery

Purpose: The purposes of this study are to explore the roles of microRNA-218 (miR-218) delivered by a newly designed magnetic nanocarrier: GCC-Fe3O4 (GCC-Fe) in dentinogenesis potentials of human dental pulp stem cells (DPSCs). Methods: Human DPSCs were obtained from impacted wisdom teeth of healthy donors under the permission of National Taiwan University Hospital institutional review board (NTUH IRB). Meanwhile, the transfection efficiency of GCC-Fe was evaluated. After transfecting miR-218 (GFm) and miR-218 inhibitor (GFmi) into DPSCs for 24 h, the dentinogenesis potentials of DPSCs were then evaluated with Alizarin Red S (ARS) staining with or without induction for 1, 4, and 9 days. Possible signaling pathway was further investigated by Western Blotting. Results: We found that the magnetic GCC-Fe3O4 nanocarrier was serum endurable with about 90% transfection efficiency in DPSCs under normal culture condition. Results of ARS staining indicated that miR-218 was negatively regulating dentinogenesis potentials of DPSCs after induction. When the miR-218 inhibitor was delivered, calcium deposits in DPSCs were increased significantly. We also discovered that the effects of miR-218 were further regulated through the MAPK/ERK pathway. Conclusion: We identified that miR-218 had a negative regulation role in the dentinogenesis of DPSCs. By inhibiting miR-218, the mineralization potentials of DPSCs were promoted after induction. In addition, we also confirmed that the highly efficient magnetic GCC-Fe3O4 nanocarrier not only was suitable for gene manipulation in biomedical studies, but also ideal for future clinical applications due to its serum endurable property. Keywords: microRNA, miR-218, Dental pulp stem cells, Dentinogenesis, Magnetic nanocarrie

Diagnostic accuracy of the CAM-ICU and ICDSC in detecting intensive care unit delirium: A bivariate meta-analysis

Background Delirium is a critical and highly prevalent problem among critically ill patients. The Confusion Assessment Method for the Intensive Care Unit (CAM-ICU) and the Intensive Care Delirium Screening Checklist (ICDSC) are the most recommended assessment tools for detecting intensive care unit (ICU) delirium. Objectives To synthesize the current evidence and compared the diagnostic accuracy of the two tools in the detection of delirium in adults in ICUs. Design Systematic review and meta-analysis. Data source A comprehensive search of the following electronic databases was performed using PubMed, Embase, CINAHL and ProQuest Dissertations and Theses A&I. The date range searched was from database inception to April 26, 2019. Review methods Two researchers independently identified articles, systematically abstracted data and evaluated the sensitivity and specificity of the CAM-ICU or the ICDSC against standard references. Bivariate diagnostic statistical analysis with a random-effects model was performed to summarize the pooled sensitivity and specificity of the two tools. Results In total, 29 CAM-ICU and 12 ICDSC studies were identified. The pooled sensitivity was 0.84 and 0.83 and pooled specificity was 0.95 and 0.87 for the CAM-ICU and the ICDSC, respectively. The CAM-ICU had higher summary specificity than the ICDSC did (p = 0.04). The percentage of hypoactive delirium, ICU type, use of mechanical ventilation, number of participants, and female percentage moderated the accuracy of the tools. Most of the domains of patient selection, index test, reference standards, and flow and timing were rated as having a low or unclear risk of bias. Conclusions Although both the CAM-ICU and the ICDSC are accurate assessment tools for screening delirium in critically ill patients, the CAM-ICU is superior in ruling out patients without ICU delirium and detecting delirium in patients in the medical ICU and those receiving mechanical ventilation. Further investigations are warranted to validate our findings

Imiquimod-induced ROS production disrupts the balance of mitochondrial dynamics and increases mitophagy in skin cancer cells

Background: Mitochondrial homeostasis is a highly dynamic process involving continuous fission and fusion cycles and mitophagy to maintain mitochondrial functionality. Imiquimod (IMQ), a Toll-like receptor (TLR) 7 ligand, is used to treat various skin malignancies. IMQ also induces apoptotic and autophagic cell death in various cancers through a TLR7-independent pathway. Objective: To investigate whether IMQ-induced ROS production is involved in mitochondrial dysfunction, mitochondrial fragmentation and mitophagy in skin cancer cells. Methods: BCC/KMC-1, B16F10 and A375 skin cancer cells, AGS gastric cancer cells and primary human keratinocytes were treated with 50 μg/mL IMQ. After 4 h, ROS were detected by CM-H2DCFDA, DHE, and MitoSOX Red staining. After 24 h, cell viability and the mitochondrial membrane potential were evaluated by a CCK-8 assay and JC-1 staining, respectively. Oxygen consumption was assessed with an Oroboros instrument. Mitochondrial morphology and mitophagy were evaluated by MitoTracker and LysoTracker staining. Mitochondrial dynamics markers, including MFN-1, DRP-1 and OPA1, and mitophagy markers, including LC3, S65-phosphorylated ubiquitin, PINK1 and TOM20, were detected by immunoblotting. Results: IMQ not only induced severe ROS production but also resulted in increased mitochondrial membrane potential loss, mitochondrial fission and mitophagy and decreased oxygen consumption in skin cancer cells compared with normal keratinocytes. Pretreatment with the antioxidant NAC reduced IMQ-induced ROS production and attenuated IMQ-induced mitochondrial fission and mitophagy in skin cancer cells. Conclusions: IMQ-induced ROS might be associated with mitochondrial dysfunction, mitochondrial fission and mitophagy in cancer cells. Alleviating IMQ-induced ROS production would reduce mitochondrial fission-to-fusion skewing and further reduce IMQ-induced mitophagy

A multicenter, randomized, open-label, controlled trial to evaluate the efficacy and tolerability of hydroxychloroquine and a retrospective study in adult patients with mild to moderate coronavirus disease 2019 (COVID-19).

ObjectiveIn this study, we evaluated the efficacy of hydroxychloroquine (HCQ) against coronavirus disease 2019 (COVID-19) via a randomized controlled trial (RCT) and a retrospective study.MethodsSubjects admitted to 11 designated public hospitals in Taiwan between April 1 and May 31, 2020, with COVID-19 diagnosis confirmed by pharyngeal real-time RT-PCR for SARS-CoV-2, were randomized at a 2:1 ratio and stratified by mild or moderate illness. HCQ (400 mg twice for 1 d or HCQ 200 mg twice daily for 6 days) was administered. Both the study and control group received standard of care (SOC). Pharyngeal swabs and sputum were collected every other day. The proportion and time to negative viral PCR were assessed on day 14. In the retrospective study, medical records were reviewed for patients admitted before March 31, 2020.ResultsThere were 33 and 37 cases in the RCT and retrospective study, respectively. In the RCT, the median times to negative rRT-PCR from randomization to hospital day 14 were 5 days (95% CI; 1, 9 days) and 10 days (95% CI; 2, 12 days) for the HCQ and SOC groups, respectively (p = 0.40). On day 14, 81.0% (17/21) and 75.0% (9/12) of the subjects in the HCQ and SOC groups, respectively, had undetected virus (p = 0.36). In the retrospective study, 12 (42.9%) in the HCQ group and 5 (55.6%) in the control group had negative rRT-PCR results on hospital day 14 (p = 0.70).ConclusionsNeither study demonstrated that HCQ shortened viral shedding in mild to moderate COVID-19 subjects