Identification and Characterization of PSEUDO-RESPONSE REGULATOR (PRR) 1a and 1b Genes by CRISPR/Cas9-Targeted Mutagenesis in Chinese Cabbage (Brassica rapa L.)

The CRISPR/Cas9 site-directed gene-editing system offers great advantages for identifying gene function and crop improvement. The circadian clock measures and conveys day length information to control rhythmic hypocotyl growth in photoperiodic conditions, to achieve optimal fitness, but operates through largely unknown mechanisms. Here, we generated core circadian clock evening components, Brassica rapa PSEUDO-RESPONSE REGULATOR (BrPRR) 1a, 1b, and 1ab (both 1a and 1b double knockout) mutants, using CRISPR/Cas9 genome editing in Chinese cabbage, where 9–16 genetic edited lines of each mutant were obtained. The targeted deep sequencing showed that each mutant had 2–4 different mutation types at the target sites in the BrPRR1a and BrPRR1b genes. To identify the functions of BrPRR1a and 1b genes, hypocotyl length, and mRNA and protein levels of core circadian clock morning components, BrCCA1 (CIRCADIAN CLOCK-ASSOCIATED 1) and BrLHY (LATE ELONGATED HYPOCOTYL) a and b were examined under light/dark cycles and continuous light conditions. The BrPRR1a and 1ab double mutants showed longer hypocotyls, lower core circadian clock morning component mRNA and protein levels, and a shorter circadian rhythm than wildtype (WT). On the other hand, the BrPRR1b mutant was not significantly different from WT. These results suggested that two paralogous genes may not be associated with the same regulatory function in Chinese cabbage. Taken together, our results demonstrated that CRISPR/Cas9 is an efficient tool for achieving targeted genome modifications and elucidating the biological functions of circadian clock genes in B. rapa, for both breeding and improvement.     

Isolation and Characterization of Urinary Extracellular Vesicles from Healthy Donors and Patients with Castration-Resistant Prostate Cancer

Prostate cancer (PCa) is the most commonly diagnosed malignancy among men in developed countries. The five-year survival rate for men diagnosed with early-stage PCa is approximately 100%, while it is less than 30% for castration-resistant PCa (CRPC). Currently, the detection of prostate-specific antigens as biomarkers for the prognosis of CRPC is criticized because of its low accuracy, high invasiveness, and high false-positive rate. Therefore, it is important to identify new biomarkers for prediction of CRPC progression. Extracellular vesicles (EVs) derived from tumors have been highlighted as potential markers for cancer diagnosis and prognosis. Specifically, urinary EVs directly reflect changes in the pathophysiological conditions of the urogenital system because it is exposed to prostatic secretions. Thus, detecting biomarkers in urinary EVs provides a promising approach for performing an accurate and non-invasive liquid biopsy for CPRC. In this study, we effectively isolated urinary EVs with low protein impurities using size-exclusion chromatography combined with ultrafiltration. After EV isolation and characterization, we evaluated the miRNAs in urinary EVs from healthy donors and patients with CRPC. The results indicated that miRNAs (miR-21-5p, miR-574-3p, and miR-6880-5p) could be used as potential biomarkers for the prognosis of CRPC. This analysis of urinary EVs contributes to the fast and convenient prognosis of diseases, including CRPC, in the clinical setting.     

Therapeutic Effects of Insulin-Producing Human Umbilical Cord-Derived Mesenchymal Stem Cells in a Type 1 Diabetes Mouse Model

In patients with type 1 diabetes (T1D), compromised pancreatic β-cell functions are compensated through daily insulin injections or the transplantation of pancreatic tissue or islet cells. However, both approaches are associated with specific challenges. The transplantation of mesenchymal stem cells (MSCs) represents a potential alternative, as MSCs have tissue-forming capacity and can be isolated from various tissues. The human umbilical cord (hUC) is a good source of freely available MSCs, which can be collected through pain-free, non-invasive methods subject to minimal ethical concerns. We sought to develop a method for the in vitro generation of insulin-producing cells (IPCs) using MSCs. We examined the potential therapeutic uses and efficacy of IPCs generated from hUC-derived MSCs (hUC-IPCs) and human adipose tissue (hAD)-derived MSCs (hAD-IPCs) through in vitro experiments and streptozotocin (STZ)-induced C57BL/6 T1D mouse models. We discovered that compared to hAD-IPCs, hUC-IPCs exhibited a superior insulin secretion capacity. Therefore, hUC-IPCs were selected as candidates for T1D cell therapy in mice. Fasting glucose and intraperitoneal glucose tolerance test levels were lower in hUC-IPC-transplanted mice than in T1D control mice and hAD-IPC-transplanted mice. Our findings support the potential use of MSCs for the treatment of T1D.     

用响应面法进行压缩机环状管*的设计优化

从结构振动和结构噪声的角度来看,压缩机的环状管是电冰箱中最重要的部件。压缩机内部机体产生的振动能量通过环状管传到壳体和外界,因此压缩机环状管的设计非常重要。但是,由于环状管几何形状的复杂性和运动的非线性,分析和设计环状管十分困难,往往不得不用统计法和试验法进行系统设计。对于类似环状管这样的复杂系统,响应面法(RSM)是常用的建模技术。用有限元模型和简单的试验模型代替环状管的实际模型作为环状管优化的起点。建立RS模型以后,用Sensitivity-based优化法进行优化。采用动态最小二乘法(MLSM)减少逼近误差。     

The real-time functional test facility for advanced instrumentationand control in nuclear power plants

Testing and validation of the functions and performance of the digital instrumentation and control (I&C) system should be done prior to installation in nuclear power plants. The objective of the I&C Functional Test Facility (FTF) is to test and validate the functions of developed digital control and various monitoring systems. The FTF provides the simulated testing environment as an experimental test bed. The FTF software consists of a mathematical modeling program which simulates a three-loop 993 MWe pressurized water reactor and a supervisory program that comprises all the instructions necessary to run the FTF. The hardware equipment provides an interface between a host computer and a simple test panel or the developed target systems to be tested. The graphical user interface supports an easy and friendly interface between the FTF and users. It is implemented through a Picasso-3 graphic tool developed by the Halden Reactor Project. The FTF is applied to an advanced I&C system prototype, such as an automatic start-up intelligent control system, dynamic alarm system, accident identification system, and intelligent logic tracking system, to test its algorithm or performance. The results of the test show good operational performance of the FTF in normal and transient conditions     

Low energy electron storage ring with tunable compaction factor

A low energy electron storage ring is designed to have many desirable properties, such as varying momentum compaction factor, damping partition numbers, favorable betatron tunes for multiturn accumulations, and excellent dynamic aperture. This storage ring can be used for debunching rf linac beams in one turn, for compression of linac pulses, and more importantly for a compact photon source based on inverse Compton scattering of laser beams.     

Kinematics, kinetics and muscle activities of the lower extremity during the first four steps from gait initiation to the steady-state walking

In this study, biomechanical characteristics during the whole process of gait initiation for twenty normal healthy volunteers were determined by the motion analysis with six near-infrared cameras, four forceplates, and an EMG system. Gait initiation, a transitional movement phenomenon from quiet stance to steady-state walking, involves a series of muscular activities, GRFs, movements of COP and COM, and joint motions. Results showed that the location of the net COP to be most lateral during double limb stance at the beginning of gait initiation. During gait initiation, changes in anteroposterior components of GRFs were first found and then changes in vertical components followed. Hip and knee motions were found before the ankle joint motion. Walking speed, step length, and stride length gradually increased until the second step. The interaction between the COM and COP is tightly regulated to control the trajectory of the COM and thereby control total body balance. This paper was recommended for publication in revised form by Associate Editor Hong Hee Yoo Sun-Woo Park received a B.S. degree in Biomedical Engineering from Yonsei University in 2006. He is currently a M.S. candidate in the Department of Biomedical Engineering, Yonsei University, Korea. His research interests in the area of Human Movement and detection of gait phase using motion sensors. Hue-Seok Choi received a B.S. degree in Computer Engineering from Daejeon University in 2004. He is currently a P.D. candidate at the Department of Biomedical Engineering at Yonsei University, Korea. His research interests are in the area of Human Movement and Rehabilitation Engineering. Ki-Hong Ryu received a B.S. and M.S. degrees in Biomedical Engineering from Yonsei University in 1998 and 2001, respectively. He is currently a P.D. candidate in the Department of Biomedical Engineering, Yonsei University, Korea. His research interests are in the area of Human Movement and Gait Training System using Functional Electrical Stimulation. Sa-Yup Kim received a B.S. degree in Electrical Engineering from Yeungnam University in 2002 and M.S. degrees in Biomedical Engineering from Yonsei University in 2006. He is currently working from Korea Institute of Industrial Technology in Cheonan, Korea. Young-Ho Kim received a B.S. degree in Mechanical Engineering from Hanyang University in 1982. He then went on to receive his M.S. and Ph.D. degrees from University of Iowa in 1989 and 1991, respectively. He is currently a Professor in the Department of Biomedical Engineering, Yonsei University, Korea. His research interests are in the area of Human Movement, Rehabilitation Engineering, and Biomechanics.     

A study on the critical speed of worn wheel profile using a scale model

A study was conducted to analyze the influence of wheel profile wear on running stability by using a 1/5 scale bogie model. The critical speed was analyzed by constructing a dynamic model for the bogie model. And the 1/5 scale bogie and a scaled roller rig prototype were manufactured to verify the feasibility of applying the scale model; a critical speed test was conducted for worn wheel profile samples. The test results for the scale model were consistent with the simulation results for the scale model within the range of 5% or less. Also, the variation of the critical speed of the scale model for the wheel profile wear shows a similar trend with the analysis results of the full scale model. Therefore, using the scale model to analyze the influence of wheel profile wear on running stability shows sufficient feasibility as a model to analyze the stability of a full scale bogie.     

Bioinformatics of Differentially Expressed Genes in Phorbol 12-Myristate 13-Acetate-Induced Megakaryocytic Differentiation of K562 Cells by Microarray Analysis

Megakaryocytes are large hematopoietic cells present in the bone marrow cavity, comprising less than 0.1% of all bone marrow cells. Despite their small number, megakaryocytes play important roles in blood coagulation, inflammatory responses, and platelet production. However, little is known about changes in gene expression during megakaryocyte maturation. Here we identified the genes whose expression was changed during K562 leukemia cell differentiation into megakaryocytes using an Affymetrix GeneChip microarray to determine the multifunctionality of megakaryocytes. K562 cells were differentiated into mature megakaryocytes by treatment for 7 days with phorbol 12-myristate 13-acetate, and a microarray was performed using RNA obtained from both types of cells. The expression of 44,629 genes was compared between K562 cells and mature megakaryocytes, and 954 differentially expressed genes (DEGs) were selected based on a p-value < 0.05 and a fold change >2. The DEGs was further functionally classified using five major megakaryocyte function-associated clusters—inflammatory response, angiogenesis, cell migration, extracellular matrix, and secretion. Furthermore, interaction analysis based on the STRING database was used to generate interactions between the proteins translated from the DEGs. This study provides information on the bioinformatics of the DEGs in mature megakaryocytes after K562 cell differentiation.