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RNA-Based dCas9–VP64 System Improves the Viability of Cryopreserved Mammalian Cells 基于rna的dCas9-VP64系统提高哺乳动物细胞低温保存活力
IF 0.8 Q4 MATERIALS SCIENCE, BIOMATERIALS Pub Date : 2018-09-01 DOI: 10.1142/S1793984418500046
Yong Hu, Lei Li, Yin Yu, Haishui Huang, B. Uygun, M. Yarmush
Regenerative therapies require availability of an abundant healthy cell source which can be achieved by efficient cryopreservation techniques. Here, we established a novel approach for improved cell cryopreservation using an mRNA-based dCas9-VP64 gene activation system for transient, yet highly efficient expression of epigenetic related genes in mammalian cells for repression of metabolic activity. Before freezing, mammalian cells were treated by dCas9-VP64-modified mRNA and guide RNAs for upregulation of histone deacetylase (HDAC), DNA methyltransferase (DNMT) and transcriptional co-repressor Sin3A genes. Cell viability, karyotype, pluripotency, and other cell specific functions were analyzed during post-thaw culture. Using conventional cryopreservation protocols, we found improvement of viability in dCas9-VP64 pretreated cells ([Formula: see text]) compared to untreated cells. Combined with dCas9-VP64 system, a reduced amount of cryoprotectant (5% DMSO) did not negatively affect the post-thaw viability. Co-delivering chemically modified dCas9-VP64 mRNA with gRNAs is an efficient gene delivery method compared to DNA-based strategies, without the associated safety concerns. This approach is a simple, yet effective way to accelerate a wide array of cellular research and translational medical applications.
再生疗法需要丰富的健康细胞来源,这可以通过有效的冷冻保存技术来实现。在这里,我们建立了一种改进细胞冷冻保存的新方法,使用基于信使核糖核酸的dCas9-VP64基因激活系统,在哺乳动物细胞中短暂但高效地表达表观遗传相关基因,以抑制代谢活性。冷冻前,用dCas9-VP64修饰的mRNA和引导RNA处理哺乳动物细胞,以上调组蛋白脱乙酰酶(HDAC)、DNA甲基转移酶(DNMT)和转录共阻遏物Sin3A基因。在解冻后培养过程中分析细胞活力、核型、多能性和其他细胞特异性功能。使用传统的冷冻保存方案,我们发现与未处理的细胞相比,dCas9-VP64预处理的细胞([公式:见正文])的生存能力有所提高。与dCas9-VP64系统相结合,冷冻保护剂(5%DMSO)的减少量不会对解冻后的生存能力产生负面影响。与基于DNA的策略相比,用gRNA共同递送化学修饰的dCas9-VP64mRNA是一种有效的基因递送方法,没有相关的安全问题。这种方法是一种简单而有效的方法,可以加速广泛的细胞研究和转化医学应用。
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引用次数: 2
Correction in Bicinchoninic Acid (BCA) Absorbance Assay to Analyze Protein Concentration 修正比辛胆酸(BCA)吸光度法分析蛋白质浓度
IF 0.8 Q4 MATERIALS SCIENCE, BIOMATERIALS Pub Date : 2018-09-01 DOI: 10.1142/S1793984418500058
Daniel Smith, Elizabeth N. Lemieux, Sutapa Barua
Conducting the bicinchoninic acid (BCA) assay directly after a coupling reaction using (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide) (EDC) and [Formula: see text]-hydroxysuccinimide (NHS) chemistry produces significant errors. Here we present a correction for the quantification of gelatin in the supernatant (SN) following gelatin conjugation to polymer microparticles using EDC and NHS chemistry. Following the conjugation reaction, SNs from the gelatin-microparticle formation reaction are treated with BCA assay reagents and quantified for the percentage of unbound gelatin in the solution. NHS was found to interfere with the BCA assay reagents and is dependent on incubation time. It is found that the large concentration (500[Formula: see text][Formula: see text]g/mL) of NHS in the conjugation reaction interferes with the sensitivity of gelatin present in SNs. The interference from NHS requires a careful analysis to distinguish the BCA background absorbance from the sample absorbance. Using an NHS control solution can correct NHS interference and thus decrease the expensive iterations in gelatin quantification and enable accurate analysis of gelatin content. The accuracy of gelatin quantification is further improved by reducing the BCA assay incubation time to approximately 20[Formula: see text]min, compared with the recommended 30[Formula: see text]min. This re-assessment of BCA assay is important to avoid misestimating biases in bioconjugation processes.
在使用(1-乙基-3-(3-二甲氨基丙基)碳二亚胺(EDC)和[公式:见正文]-羟基琥珀酰亚胺(NHS)化学偶联反应后直接进行bicinchoninic酸(BCA)测定会产生显着误差。在这里,我们提出了明胶定量上清(SN)后明胶偶联到聚合物微粒使用EDC和NHS化学校正。在结合反应之后,用BCA测定试剂处理明胶-微粒形成反应产生的SNs,并定量测定溶液中未结合明胶的百分比。发现NHS干扰BCA测定试剂,并依赖于孵育时间。结果发现,偶联反应中NHS的大浓度(500 g/mL)干扰了SNs中明胶的敏感性。来自NHS的干扰需要仔细分析,以区分BCA背景吸光度与样品吸光度。使用NHS控制解决方案可以纠正NHS干扰,从而减少明胶定量中昂贵的迭代,并能够准确分析明胶含量。与推荐的30分钟相比,将BCA测定的孵育时间减少到大约20分钟,进一步提高了明胶定量的准确性。对BCA检测的重新评估对于避免生物偶联过程中的错误估计偏差很重要。
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引用次数: 5
Nano Engineering of Artificial Granulocytes for Cancer Diagnosis and Therapeutics 用于癌症诊断和治疗的人造粒细胞纳米工程
IF 0.8 Q4 MATERIALS SCIENCE, BIOMATERIALS Pub Date : 2018-09-01 DOI: 10.1142/S1793984418710010
Donglu Shi
The study of nanomedicine research, to date, has been concentrating on developing nanovectors for medical imaging, drug/gene delivery, and cell targeting, particularly in cancer diagnosis and therapeutics. Although quite successful in some areas, critical challenges remain, especially in the clinical settings. Some of the critical issues include delivery inefficiency, targeting non-specificity, and low uptake of theranostic-payloads by the cancerous lesions upon intravenous injection. Therefore, an alternative approach may be possible, via nanotechnology, by simulating some of the immune cells. In this Technical Note, we propose using specifically designed nanoparticles to simulate neutrophils that are capable of effective cancer cells targeting and killing without the complications in drug loading, biomarker conjugation, and assembly of chemical and physical therapeutic means. The simulated artificial cells mimic neutrophils, namely, granulocytes, according to their biological characteristics, for instance, positively-charged cell surfaces and the ability to release perferin upon binding onto the cancer cells leading to cytolysis. This Technical Note is intended to deliver a new concept in artificially-engineered granulocytes for cancer diagnosis and therapeutics.
迄今为止,纳米医学研究的重点是开发用于医学成像、药物/基因递送和细胞靶向的纳米载体,特别是在癌症诊断和治疗中。尽管在某些领域相当成功,但仍然存在重大挑战,尤其是在临床环境中。一些关键问题包括递送效率低、靶向非特异性和静脉注射时癌性病变对治疗有效载荷的吸收低。因此,通过纳米技术,通过模拟一些免疫细胞,一种替代方法可能是可行的。在本技术说明中,我们建议使用专门设计的纳米颗粒来模拟中性粒细胞,这些中性粒细胞能够有效靶向和杀死癌症细胞,而不会出现药物负载、生物标志物结合以及化学和物理治疗手段组装方面的并发症。模拟的人工细胞模仿中性粒细胞,即粒细胞,根据它们的生物学特征,例如,带正电荷的细胞表面和结合到癌症细胞上释放perferin导致细胞溶解的能力。本技术说明旨在为癌症诊断和治疗提供人工工程粒细胞的新概念。
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引用次数: 0
Residential Satisfaction of Elderly as Determinant Behind Design Thinking in Urban Planning 老年人居住满意度:城市规划设计思维的决定因素
IF 0.8 Q4 MATERIALS SCIENCE, BIOMATERIALS Pub Date : 2018-06-01 DOI: 10.1142/S1793984418400044
Yi-ling Yu, Fuqin Wang, Feiyang Zhu, Zhigang Yin
Residential satisfaction (RS) is an important bolometer to capture the mood of residents and their opinions on the quality of life they are living. Here, it is our intent to focus on the evaluation of the home as the measurement for satisfaction of the occupant, as it occurs naturally for its occupants. In this case, we study the urban folks, in terms of density, location and housing type, designed by the urban planners. Using data from the 4th National Sample Survey on the living conditions of China’s urban and rural older population, we analyzed the RS among people in their 60[Formula: see text]s and older ([Formula: see text]) living in the suburban neighborhoods in Shanghai. Using Logistic Regression models, we compared and studied the impacts of these three independent variables on their view on RS. The results confirmed the housing type and the location as the two most impactful factors and as the two main determinants in RS among the elderly population living in the urban areas of Shanghai.
居住满意度(RS)是反映居民情绪和他们对生活质量的看法的重要指标。在这里,我们的目的是关注住宅的评估,作为居住者满意度的衡量标准,因为它对居住者来说是自然发生的。在这种情况下,我们研究城市人口,根据密度,位置和住房类型,由城市规划者设计。利用第四次全国城乡老年人口生活状况抽样调查数据,对上海郊区60岁以上(公式:见文本)老年人的RS进行了分析。采用Logistic回归模型,比较研究了这三个自变量对老年人生活满意度的影响,结果证实住房类型和地理位置是影响上海城区老年人生活满意度的两个最主要因素和两个主要决定因素。
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引用次数: 4
Spatial Clustering of Pedestrian Activity and the Built Environment Characteristics 行人活动空间聚类与建成环境特征
IF 0.8 Q4 MATERIALS SCIENCE, BIOMATERIALS Pub Date : 2018-06-01 DOI: 10.1142/S1793984418400056
Haotong Zhang, L. Yin
Promoting pedestrian activity has attracted increasing attention as an important strategy for the improvement of public health and urban revitalization. The impact on physical activity underpinned by built environment has been studied substantially; however, few studies had focused on the geographically varying relationships between pedestrian activity and the built environment characteristics. Built upon previous work, this study looks at the spatial patterns of pedestrian counts and the built environment contributors along two major streets in Buffalo, New York using global and local spatial autocorrelation tests and geographically weighted regression. Pedestrian generators, job density and land use mix are included as independent variables in order to study the impact on them due to the characteristics of built environment. Our findings suggest that (1) there are statistically significant clusters of street intersections with high pedestrian counts along the streets selected in our study; (2) there are some optimal sizes of clusters of pedestrian generators, which attract more pedestrians; (3) geographically weighted Poisson model helps to analyze the geographically varying relationships between the built environment and pedestrian activity with a more pronounced goodness of fit. This research contributes to the understanding of the spatial patterns of pedestrian activity and the geographically varying relationship between the built environment and pedestrian counts. Hopefully this research will help to guide and focus the minds of policy makers and urban planners alike to introduce street vitality through the modifications of the built environment, so as to improve the quality of life in their neighborhoods.
促进步行活动作为改善公共卫生和城市振兴的一项重要战略,越来越受到人们的关注。已对建筑环境对身体活动的影响进行了大量研究;然而,很少有研究关注行人活动与建成环境特征之间的地理变化关系。在先前工作的基础上,本研究使用全球和本地空间自相关测试和地理加权回归,研究了纽约布法罗两条主要街道上行人数量和建筑环境贡献者的空间模式。将行人产生量、工作密度和土地利用组合作为自变量,研究建筑环境特征对其的影响。研究结果表明:(1)所选街道沿线存在显著的行人数量较多的十字路口集群;(2)行人发生器集群存在一定的最优规模,能够吸引更多的行人;(3)地理加权泊松模型有助于分析建成环境与行人活动之间的地理变化关系,拟合优度更为显著。该研究有助于理解行人活动的空间格局以及建筑环境与行人数量之间的地理变化关系。希望这项研究能够帮助政策制定者和城市规划者通过改造建筑环境来引入街道活力,从而提高社区的生活质量。
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引用次数: 0
Impact of Built Environment on Respiratory Health: An Empirical Study 建筑环境对呼吸系统健康影响的实证研究
IF 0.8 Q4 MATERIALS SCIENCE, BIOMATERIALS Pub Date : 2018-06-01 DOI: 10.1142/S1793984418400019
Wangyue Xu, Xiaojing Zhao, Lan Wang
In many heavy industrialized countries such as China and India, the impact of air pollution on respiratory health has been headline news in recent years. Among the risk issues examined, exposure to particulate matter (PM) is cited as the prime contributing factor that causes respiratory diseases, yet it is traceable and controllable. In this paper, we report on an empirical study conducted in Shanghai, based on urban spatial determinants as independent variables to investigate its link to occurrence of lung cancer in their neighborhoods. A survey was conducted on a population of 472 lung cancer patients. After reliability and validity tests, only 156 pairs were included in this report. The questionnaire designed for this survey covers 11 outdoor and 6 indoor factors; these include the building density where they live, proximity to pollution sources, volume of traffic nearby, degree of enclosure by surrounding tall buildings, being residential or commercial with reference to their homes, proximity to parks, measured in terms of the plant type, green space per capita, accessibility to public open spaces for outdoor activities and water body; while parameters inside the house cover the age of the house, bedroom sizes, space per occupant, cooling-off time of taking up residency after renovation, humidity and dust inside the houses. Data collected were classified using random forest classification and further refined with Boruta algorithm for feature selection to identify possible correlation between risk of lung cancer to both outdoor and indoor factors of built environment. The results reveal a strong correlation between lung cancer and the environment where they live, so much so that the finding confirms our long-held belief that urban redevelopment could play an important role in reducing the risk of respiratory disease. Since prevention is better than cure, if by design to relocate pollution sources away from residential areas, provision of good public transportation to cut down vehicles on our streets, creation of green spaces to improve airflow pathway to deduce the concentration of PM in the atmosphere in our neighborhoods, we could perhaps reduce or even prevent lung cancer and a range of other respiratory diseases for the residents we served.
在中国和印度等许多重工业化国家,空气污染对呼吸系统健康的影响近年来一直是头条新闻。在检查的风险问题中,暴露于颗粒物(PM)被认为是导致呼吸道疾病的主要因素,但它是可追踪和可控的。在本文中,我们报道了在上海进行的一项实证研究,以城市空间决定因素为自变量,研究其与社区癌症发生的关系。对472名癌症患者进行了调查。经过可靠性和有效性测试,本报告仅包括156对。为本次调查设计的问卷包括11个室外因素和6个室内因素;这些因素包括他们居住的建筑密度、污染源的接近程度、附近的交通量、周围高层建筑的封闭程度(相对于他们的住宅或商业建筑)、公园的接近程度(按植物类型、人均绿地、户外活动公共开放空间的可达性和水体衡量);而房子内部的参数包括房子的年龄、卧室大小、每个居住者的空间、装修后居住的冷却时间、房子内部的湿度和灰尘。使用随机森林分类对收集的数据进行分类,并使用Boruta算法进一步细化特征选择,以识别癌症风险与建筑环境的室外和室内因素之间的可能相关性。研究结果表明,癌症与他们生活的环境之间存在着强烈的相关性,这一发现证实了我们长期以来的信念,即城市重建可以在降低呼吸道疾病风险方面发挥重要作用。由于预防胜于治疗,如果通过设计将污染源从居民区转移,提供良好的公共交通来减少街道上的车辆,创建绿色空间来改善气流通道,以降低我们社区大气中PM的浓度,我们也许可以为我们服务的居民减少甚至预防肺癌和一系列其他呼吸道疾病。
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引用次数: 3
Ethnography for Accessibility to Public Open Spaces 公共开放空间的可达性人种学
IF 0.8 Q4 MATERIALS SCIENCE, BIOMATERIALS Pub Date : 2018-06-01 DOI: 10.1142/S1793984418400020
Yu-lin Lu, H. Leng, Qing Yuan
The access to public open spaces (POSs) has been catching the attention of urban planners and architects alike to inject dynamism into their design so as to bring life back into the static structures they have or about to create. POSs are not just green spaces existing between buildings, but spaces to host events, grounds for physical exercise, as well as rehabilitation workout places for patients discharged from the hospitals. In China, 44.6% of the country, mostly located in high latitude and altitude areas, accessibility to POSs is often hampered by bad weather. In this paper, we reported on an ethnography process from the point of view of pedestrians walking in the cold to experience the accessibility to POSs using Global Position System (GPS) mobile phones carried in their pockets, as a biosensor, to study the behaviors of pedestrians on the move, so as to identify the access points for possible urban redevelopment, where weatherproof linkways could be put in place to facilitate and encourage people to venture out into the open during winter months. In the course of our research, we undertook a case study in the central districts of Harbin that includes Nangang, Daoli, Daowai and Xiangfang districts, during the winter months (from December 2015 to June 2016), enduring an average monthly temperature of [Formula: see text]C, to ethnography, the pattern of pedestrians walking in the cold. The results confirm our hypothesis that harsh cold environment and the extreme climate conditions have discouraged the access to POSs. At the same time, the data collected have also helped us to pinpoint access points where weatherproof link way could be provided to facilitate and encourage people to travel by foot during winter months.
进入公共开放空间(POS)一直吸引着城市规划者和建筑师的注意,为他们的设计注入活力,从而使他们拥有或即将创建的静态结构恢复活力。POS不仅仅是建筑物之间的绿地,也是举办活动的空间、体育锻炼的场地,以及出院患者的康复锻炼场所。在中国,44.6%的地区大多位于高纬度和高海拔地区,恶劣天气经常阻碍POS的使用。在本文中,我们报道了一个民族志过程,从行人在寒冷中行走的角度出发,使用口袋里携带的全球定位系统(GPS)手机作为生物传感器来体验POS的可达性,以研究行人在移动中的行为,从而确定可能的城市重建的接入点,在那里可以设置防风雨的连接道,以方便和鼓励人们在冬季冒险外出。在我们的研究过程中,我们在哈尔滨的中心区进行了一个案例研究,包括南岗、道里、道外和香坊区,在冬季(2015年12月至2016年6月),忍受着[公式:见正文]C的月平均温度,民族志,行人在寒冷中行走的模式。研究结果证实了我们的假设,即恶劣的寒冷环境和极端的气候条件阻碍了获得POS。与此同时,收集到的数据也帮助我们确定了可以提供防风雨连接方式的接入点,以方便和鼓励人们在冬季徒步旅行。
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引用次数: 2
Towards Pedestrian Microclimatic Comfort: A Rapid Predication Model for Street Winds and Pedestrian Thermal Sensation 行人微气候舒适度:街道风和行人热感觉的快速预测模型
IF 0.8 Q4 MATERIALS SCIENCE, BIOMATERIALS Pub Date : 2018-06-01 DOI: 10.1142/S1793984418400068
Jing Li, MengNan Qi, Qiuhua Duan, Lei Huo, Julian Wang
Significant changes in the urban built environment have occurred due to rapid urbanization and increases in the urban population. Such alterations may produce environmental health-related issues such as urban heat stress, air pollution and traffic noise. This research undertook a field study to collect data including urban design parameters, micro-environmental factors and city climatic information. This work was conducted over a two-year period on three pedestrian streets located in high-density urban areas in Beijing. These areas were selected in order to study the influences of urban street canyon texture within a particular geometric layout, wind flow corridors and variations in air temperature on pedestrian microclimatic comfort. The results will facilitate the work of urban planners by providing them with information for use in improving outdoor thermal comfort through their designs. A total of 60[Formula: see text]485 samples were organized into training, validation and test sets. We confirmed our hypothesis that internal wind speed ([Formula: see text] is attributable mainly to the urban texture coefficient ([Formula: see text], air temperature ([Formula: see text] and leading-in wind speed ([Formula: see text]. The model was tested using the test data collected onsite, which demonstrated a very accurate goodness-of-fit; the model achieved an R-squared value of 0.82, which meant that [Formula: see text] as a dependent variable was 82% correlated to the three predictors as independent variables. With this computer simulation, urban planners can now predict and visualize the impact of changes on the built environment in terms of either the direction of solar radiation received or increases in wind speed, in return for the desired thermal comfort level for residents of the neighborhood.
由于城市化的快速发展和城市人口的增加,城市建筑环境发生了重大变化。这种改变可能产生与环境有关的问题,如城市热应激、空气污染和交通噪音。本研究通过实地调研,收集了城市设计参数、微环境因子、城市气候信息等数据。这项工作在北京高密度城区的三条步行街进行了为期两年的研究。选择这些区域是为了研究特定几何布局下的城市街道峡谷纹理、风流道和气温变化对行人微气候舒适度的影响。研究结果将为城市规划者的工作提供便利,为他们通过设计改善室外热舒适提供信息。共60个[公式:见文]485个样本被组织成训练集、验证集和测试集。我们证实了我们的假设,即内部风速([公式:见文])主要归因于城市肌理系数([公式:见文])、气温([公式:见文])和引入风速([公式:见文])。利用现场采集的测试数据对模型进行了测试,得到了非常准确的拟合优度;模型的r平方值为0.82,这意味着[公式:见文本]作为因变量与三个预测因子作为自变量的相关性为82%。有了这个计算机模拟,城市规划者现在可以预测和可视化变化对建筑环境的影响,无论是接收太阳辐射的方向还是风速的增加,以换取社区居民所需的热舒适水平。
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引用次数: 0
Preface 前言
IF 0.8 Q4 MATERIALS SCIENCE, BIOMATERIALS Pub Date : 2018-06-01 DOI: 10.1142/s1793984418020014
Lanqin Wang
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引用次数: 0
Cardiovascular Diseases Due to Stress Arisen from Social Risk Factors: A Synopsis and Prospectiveness 社会风险因素引起的应激性心血管疾病:综述与展望
IF 0.8 Q4 MATERIALS SCIENCE, BIOMATERIALS Pub Date : 2018-06-01 DOI: 10.1142/S1793984418400032
Bin Jiang, Jielin Chen, Lan Wang
This paper reviews the causes of cardiovascular diseases (CVD) from a perspective other than traditional clinical pathology. We look at social factors that could arouse stressful feelings, and could further lead to cardiovascular diseases. Major social factors including job stress, daily hassles, life events, social inequity and trauma are covered in our review. This paper also discusses potential interventions which could be taken by environmental professionals to create dynamism within the existing static structures so as to eventually eradicate the causes of mental stress and ameliorate the quality of life for urban dwellers.
本文从传统临床病理学以外的角度综述了心血管疾病(CVD)的病因。我们关注的是可能引发压力感,并可能进一步导致心血管疾病的社会因素。我们的综述涵盖了主要的社会因素,包括工作压力、日常烦恼、生活事件、社会不平等和创伤。本文还讨论了环境专业人员可以采取的潜在干预措施,以在现有的静态结构中创造活力,从而最终消除心理压力的原因,改善城市居民的生活质量。
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引用次数: 0
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