Generation of Multicellular Tumor Spheroids with Microwell-Based Agarose Scaffolds for Drug Testing

Three dimensional multicellular aggregate, also referred to as cell spheroid or microtissue, is an indispensable tool for in vitro evaluating antitumor activity and drug efficacy. Compared with classical cellular monolayer, multicellular tumor spheroid (MCTS) offers a more rational platform to predict in vivo drug efficacy and toxicity. Nevertheless, traditional processing methods such as plastic dish culture with nonadhesive surfaces are regularly time-consuming, laborious and difficult to provide uniform-sized spheroids, thus causing poor reproducibility of experimental data and impeding high-throughput drug screening. In order to provide a robust and effective platform for in vitro drug evaluation, we present an agarose scaffold prepared with the template containing uniform-sized micro-wells in commercially available cell culture plates. The agarose scaffold allows for good adjustment of MCTS size and large-scale production of MCTS. Transparent agarose scaffold also allows for monitoring of spheroid formation under an optical microscopy. The formation of MCTS from MCF-7 cells was prepared using different-size-well templates and systematically investigated in terms of spheroid growth curve, circularity, and cell viability. The doxorubicin cytotoxicity against MCF-7 spheroid and MCF-7 monolayer cells was compared. The drug penetration behavior, cell cycle distribution, cell apoptosis, and gene expression were also evaluated in MCF-7 spheroid. The findings of this study indicate that, compared with cellular monolayer, MCTS provides a valuable platform for the assessment of therapeutic candidates in an in vivo-mimic microenvironment, and thus has great potential for use in drug discovery and tumor biology research

Concepts, Structure and Developments of High-Reliability Cyber-Physical Fusion Based Coordinated Planning for Distribution System

Coordinated control is imperative for the distribution network with the integration of wind power, photovoltaic system, and energy storage system. Meanwhile, the advanced automation terminal, intelligent control technology, and information communication technologies have greatly promoted the informatization of distribution networks which also increase the correlation between the physical system (primary system) and the cyber system (secondary system). Hence, it is critical to comprehensively coordinate the planning of the cyber-physical system for building a highly reliable power grid. This work summarizes a series of challenges brought by the highly coupled cyber-physical system, such as the primary and secondary collaborated planning models and solution algorithms. Then, the reliability assessment theories of cyber-physical systems and their application in distribution network planning models are introduced. Finally, three development directions of distribution network planning in the future are proposed, considering primary and secondary system coordinated planning

Safety risk assessment of subway shield construction under-crossing a river using CFA and FER

Numerous subway projects are planned by China's city governments, and more subways can hardly avoid under-crossing rivers. While often being located in complex natural and social environments, subway shield construction under-crossing a river (SSCUR) is more susceptible to safety accidents, causing substantial casualties, and monetary losses. Therefore, there is an urgent need to investigate safety risks during SSCUR. The paper identified the safety risks during SSCUR by using a literature review and experts' evaluation, proposed a new safety risk assessment model by integrating confirmatory factor analysis (CFA) and fuzzy evidence reasoning (FER), and then selected a project to validate the feasibility of the proposed model. Research results show that (a) a safety risk list of SSCUR was identified, including 5 first-level safety risks and 38 second-level safety risks; (b) the proposed safety risk assessment model can be used to assess the safety risk of SSCUR; (c) safety inspection, safety organization and duty, quicksand layer, and high-pressure phreatic water were the high-level risks, and the onsite total safety risk was at the medium level; (d) management-type safety risks, environment-type safety risks, and personnel-type safety risks have higher expected utility values, and manager-type safety risks were expected have higher risk-utility values when compared to worker-type safety risks. The research can enrich the theoretical knowledge of SSCUR safety risk assessment and provide references to safety managers for conducting scientific and effective safety management on the construction site when a subway crosses under a river

Coordinated Planning and Energy Conservation for Distribution Network with Renewable Energy: Standardized Information Model and Software

In recent years, energy conservation and environmental protection have attracted great attention by the state, and many efforts have been made from the policy and planning level. In view of the current distribution network planning requirements about energy-saving and environmental protection attributes such as loss reduction, carbon reduction, and environmental friendliness, this study proposes a set of energy-saving and environmental protection evaluation indicators for distribution network. Then, the CIM file library is constructed for typical equipment. Based on the CIM file, the digital planning technology for distribution network is designed. Besides, the feature library of energy conservation and environmental protection indicators, power flow calculation module, carbon flow calculation module, and renewable energy integration planning module are described

<i>Mir505–3p</i> regulates axonal development via inhibiting the autophagy pathway by targeting <i>Atg12</i>

<p>In addition to the canonical role in protein homeostasis, autophagy has recently been found to be involved in axonal dystrophy and neurodegeneration. Whether autophagy may also be involved in neural development remains largely unclear. Here we report that <i>Mir505–3p</i> is a crucial regulator for axonal elongation and branching in vitro and in vivo, through modulating autophagy in neurons. We identify that the key target gene of <i>Mir505–3p</i> in neurons is <i>Atg12</i>, encoding ATG12 (autophagy-related 12) which is an essential component of the autophagy machinery during the initiation and expansion steps of autophagosome formation. Importantly, axonal development is compromised in brains of <i>mir505</i> knockout mice, in which autophagy signaling and formation of autophagosomes are consistently enhanced. These results define <i>Mir505–3p</i>-ATG12 as a vital signaling cascade for axonal development via the autophagy pathway, further suggesting the critical role of autophagy in neural development.</p

Table_1_Safety risk assessment of subway shield construction under-crossing a river using CFA and FER.pdf

Numerous subway projects are planned by China's city governments, and more subways can hardly avoid under-crossing rivers. While often being located in complex natural and social environments, subway shield construction under-crossing a river (SSCUR) is more susceptible to safety accidents, causing substantial casualties, and monetary losses. Therefore, there is an urgent need to investigate safety risks during SSCUR. The paper identified the safety risks during SSCUR by using a literature review and experts' evaluation, proposed a new safety risk assessment model by integrating confirmatory factor analysis (CFA) and fuzzy evidence reasoning (FER), and then selected a project to validate the feasibility of the proposed model. Research results show that (a) a safety risk list of SSCUR was identified, including 5 first-level safety risks and 38 second-level safety risks; (b) the proposed safety risk assessment model can be used to assess the safety risk of SSCUR; (c) safety inspection, safety organization and duty, quicksand layer, and high-pressure phreatic water were the high-level risks, and the onsite total safety risk was at the medium level; (d) management-type safety risks, environment-type safety risks, and personnel-type safety risks have higher expected utility values, and manager-type safety risks were expected have higher risk-utility values when compared to worker-type safety risks. The research can enrich the theoretical knowledge of SSCUR safety risk assessment and provide references to safety managers for conducting scientific and effective safety management on the construction site when a subway crosses under a river.</p

Metabolomics analyses of serum metabolites perturbations associated with Naja atra bite.

Naja atra bite is one of the most common severe snakebites in emergency departments. Unfortunately, the pathophysiological changes caused by Naja atra bite are unclear due to the lack of good animal models. In this study, an animal model of Naja atra bite in Guangxi Bama miniature pigs was established by intramuscular injection at 2 mg/kg of Naja atra venom, and serum metabolites were systematically analyzed using untargeted metabolomic and targeted metabolomic approaches. Untargeted metabolomic analysis revealed that 5045 chromatographic peaks were obtained in ESI+ and 3871 chromatographic peaks were obtained in ESI-. Screening in ESI+ modes and ESI- modes identified 22 and 36 differential metabolites compared to controls. The presence of 8 core metabolites of glutamine, arginine, proline, leucine, phenylalanine, inosine, thymidine and hippuric acid in the process of Naja atra bite was verified by targeted metabolomics significant difference (P<0.05). At the same time, during the verification process of the serum clinical samples with Naja atra bite, we found that the contents of three metabolites of proline, phenylalanine and inosine in the serum of the patients were significantly different from those of the normal human serum (P<0.05). By conducting functional analysis of core and metabolic pathway analysis, we revealed a potential correlation between changes in key metabolites after the Naja atra bite and the resulting pathophysiological alterations, and our research aims to establish a theoretical foundation for the prompt diagnosis and treatment of Naja atra bite

The primer sequences used for qRT-PCR analyses.

<p>The primer sequences used for qRT-PCR analyses.</p

Assessment of drug cytotoxicity on agarose scaffolds with micro-wells.

<p>Data were expressed as mean ± standard deviation (SD) (n = 6). (A) Cytotoxicity testing of doxorubicin against MCF-7 cells in 3-D and 2-D cell culture models. (B) Size-dependent drug responses in 3-D cell culture models.</p