一种具有高信噪比的脉搏波光电传感器的研制

目的：探讨研制一种生理信号光电传感器。方法：利用光电传感器检测通过指端毛细血管后的光强度的变化，将光信号转换成电信号，再经过电信号1/V转换、放大、滤波处理。结果：获得了清晰稳定的指端容积脉搏波。结论：高信噪比的光电传感器可用作于临床监测和生理信号分析处理系统。     

城市公共厕所发展趋势的探讨

介绍了改革开放以来各地出现的一些新型公厕,认为前店后厕、附属式公厕、生态公厕是发展趋势,而豪华型公厕的推广目前尚不具备条件.     

大鼠脑干神经元型一氧化氮合酶免疫阳性神经元的分布

目的　观察大鼠脑干神经元型一氧化氮合酶 (nNOS)免疫阳性神经元的分布 ,为探讨nNOS的作用提供形态学资料。方法　用ABC免疫细胞化学方法显示脑干nNOS免疫阳性神经元。结果　大鼠脑干nNOS免疫阳性神经元以中脑和脑桥分布丰富 ,延髓较稀少 ;在中脑 ,nNOS免疫阳性神经元主要分布于中脑水管周围灰质的背侧部、被盖背外侧核、中缝背核、上下丘灰质等部位 ;在脑桥 ,主要分布于被盖背外侧核、脑桥中缝核、被盖脚桥核、蓝斑、臂旁核、斜方体核 ,以及脑桥网状结构 ;与中脑和脑桥相比 ,延髓nNOS免疫阳性神经元较少 ,主要分布于延髓网状结构、三叉神经脊束核和孤束核等核团。结论　分布于脑干内丰富的nNOS免疫阳性神经元可能通过其生成的NO调节其他神经递质的分泌 ,共同参与内脏活动、感觉和运动的传导 ,以及睡眠和觉醒等脑的高级整合功能的调节。     

Apoptosis, proliferation and gene expression patterns in mouse developing tongue

The Fgf/Fgfr (Fgf receptor) and Bmp signal pathways are critical for embryonic development and postnatal growth. In order to address their roles in tongue development, preliminary study of expression patterns of some important members in the two families, as well as of apoptosis and proliferation, were carried out in mouse developing tongue. Apoptosis in tongue is a very late event in embryogenesis, restricted to the upper layer of the epithelium whereas proliferation is very vigorous at the early stage of tongue development and remains active throughout embryogenesis. Bmp2, −4 and -5 were localized within the mesenchyme at the early embryonic stage of tongue development (E12 to E13), whereas Bmp3 and Bmp7 were mainly expressed in the epithelium. Most of these molecules were also seen in the tongue muscles at postnatal stages. Among Fgfr isoforms, Fgfr1c, −2b, and -2c were detected in embryogenesis with peak expression at E11 to E13. Fgfr1c and Fgfr2c were localized within the mesenchyme, while Fgfr2b was mainly expressed in the epithelium. High expression of Fgf7 and Fgf10 was also detected in the mesenchyme at the early embryonic stage of tongue development, corresponding to the Fgfr expression, suggesting that they are among the principal ligands functioning at the early embryonic expanding stage. Fgf2 was seen in the tongue muscles at the late embryonic and postnatal stages. These results suggest that Bmp and Fgf signalling regulates tongue development at multiple stages, possibly related to proliferation and differentiation.     

Neuronal activity related to the visual representation of arm movements in the lateral cerebellar cortex

Testing the hypothesis that the lateral cerebellum forms a sensory representation of arm movements, we investigated cortical neuronal activity in two monkeys performing visually guided step-tracking movements with a manipulandum. A virtual target and cursor image were viewed co-planar with the manipulandum. In the normal task, manipulandum and cursor moved in the same direction; in the mirror task, the cursor was left-right reversed. In one monkey, 70- and 200-ms time delays were introduced on cursor movement. Significant task-related activity was recorded in 31 cells in one animal and 142 cells in the second: 10.2% increased activity before arm movements onset, 77.1% during arm movement, and 12.7% after the new position was reached. To test for neural representation of the visual outcome of movement, firing rate modulation was compared in normal and mirror step-tracking. Most task-related neurons (68%) showed no significant directional modulation. Of 70 directionally sensitive cells, almost one-half (n = 34, 48%) modulated firing with a consistent cursor movement direction, many fewer responding to the manipulandum direction (n = 9, 13%). For those "cursor-related" cells tested with delayed cursor movement, increased activity onset was time-locked to arm movement and not cursor movement, but activation duration was extended by an amount similar to the applied delay. Hence, activity returned to baseline about when the delayed cursor reached the target. We conclude that many cells in the lateral cerebellar cortex signaled the direction of cursor movement during active step-tracking. Such a predictive representation of the arm movement could be used in the guidance of visuo-motor actions.     

A HIT-trapping strategy for rapid generation of reversible and conditional alleles using a universal donor

Targeted mutagenesis in model organisms is key for gene functional annotation and biomedical research. Despite technological advances in gene editing by the CRISPR-Cas9 systems, rapid and efficient introduction of site-directed mutations remains a challenge in large animal models. Here, we developed a robust and flexible insertional mutagenesis strategy, homology-independent targeted trapping (HIT-trapping), which is generic and can efficiently target-trap an endogenous gene of interest independent of homology arm and embryonic stem cells. Further optimization and equipping the HIT-trap donor with a site-specific DNA inversion mechanism enabled one-step generation of reversible and conditional alleles in a single experiment. As a proof of concept, we successfully created mutant alleles for 21 disease-related genes in primary porcine fibroblasts with an average knock-in frequency of 53.2%, a great improvement over previous approaches. The versatile HIT-trapping strategy presented here is expected to simplify the targeted generation of mutant alleles and facilitate large-scale mutagenesis in large mammals such as pigs.

Following the completion of animal genome sequencing projects, rapid and efficient mutagenesis strategies are needed for analyzing gene function and for creating human disease models. Gene trapping is a high-throughput mutagenesis strategy whereby random vector insertion can be achieved across the mouse genome. A typical gene-trap vector contains a promoter-less reporter/selection gene flanked by an upstream splice acceptor (SA) and a downstream poly(A) signal. Upon insertion into an intron of a gene, the vector both inactivates the trapped gene and enables the gene-specific expression of a reporter gene (Gossler et al. 1989; Stanford et al. 2001). To date, gene-trapping approaches have been successfully applied toward large-scale mutagenesis in mouse embryonic stem cells (mESCs) and generation of gene knockout mice (Skarnes et al. 2004). The main drawback of random gene trapping is that gene-trap alleles are not specifically engineered to target genes of interest in advance. Therefore, methods to streamline the introduction of predesigned, site-specific modifications into the genome by homologous recombination would represent a significant technological advance. Previously, a hybrid approach combining gene targeting and gene trapping (targeted trapping) enabled mutation of expressed genes in mESCs with high efficiency, using a gene-trap construct flanked by homologous sequences of the target locus (Friedel et al. 2005). Also, homologous recombination is commonly used for creating conditional alleles, which is essential to avoid embryonic lethality and to study the stage- and tissue-specific functions of genes (Branda and Dymecki 2004). However, both standard gene trapping and targeted trapping are only suitable for genes expressed in embryonic stem (ES) cells. Furthermore, construction of targeting donor vectors with homology arms is labor intensive and costly, and the low efficiency of homologous recombination is also a rate-limiting step for gene targeting in mammalian genomes.Recently, by taking advantage of precise genomic double-strand breaks (DSBs) created by the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) system (Ran et al. 2013; Doudna and Charpentier 2014; Hsu et al. 2014), homology-directed repair (HDR) efficiency was substantially enhanced (Porteus and Carroll 2005), and even donors with short homology arms (Orlando et al. 2010) or single-stranded DNA oligonucleotides (Chen et al. 2011; Quadros et al. 2017) were found to be compatible with site-specific integration. However, each targeting donor for HDR still needs to be customized with gene-specific homology sequences. Because of the lack of ES cells for certain animals such as pigs, sheep, and cattle, the genome must be edited either in a zygote embryo or in a somatic cell for somatic cell nuclear transfer (SCNT) (Reddy et al. 2020). It is still not feasible to achieve large-scale insertional mutagenesis including conditional knockouts in these important species with random gene trapping or HDR-based methods. Also, the problem of genetic mosaicism in embryo editing remains unresolved (Mehravar et al. 2019), prompting a need for technological advances to accelerate genetic modification in somatic cells.Alternatively, the generally more efficient nonhomologous end joining (NHEJ) pathway has been exploited for site-specific insertion of exogenous DNA by simultaneous cleavage of both donor plasmid and genome using programmable nucleases (Cristea et al. 2013; Maresca et al. 2013; Brown et al. 2016; Suzuki et al. 2016; Sawatsubashi et al. 2018). In contrast to HDR-based strategies, NHEJ-mediated insertions do not require gene-specific homology arms, enabling diverse sites to be targeted with a universal donor vector. Therefore, we speculated that a gene-trap cassette could be inserted into a specific locus easily through this mechanism in any cell type.Here, by combining NHEJ-mediated knock-in and gene trapping, we developed a strategy for targeted mutagenesis, especially in somatic cells with low HDR activity, referred to as HIT-trapping. By using a universal donor, this strategy allows us to (1) create null alleles, (2) produce a fluorescent reporter signal that could potentially allow cells with null alleles to be identified very quickly, and (3) produce reversible and conditional alleles that would be very helpful to have in most animal models but are often cumbersome to create.     

基于TRPV1/TRPA1调控TLR/NLRP3通路探讨中医药干预动脉粥样硬化研究进展

动脉粥样硬化（AS）是一种脂质堆积、血管内皮功能障碍导致的慢性炎症性疾病,Toll样受体（TLR）/核转录因子-κB(NF-κB)通路与NOD样受体蛋白3(NLRP3)炎性小体通路在动脉粥样硬化的产生中发挥着重要的致炎作用,而瞬时受体电位香草酸亚型1(TRPV1)与瞬时受体电位锚蛋白1(TRPA1)在动脉粥样硬化的发生过程中发挥着重要的保护作用。作者对近10年国内外期刊发表的相关研究进行梳理,研究显示,TRPV1/TRPA1的激活可以通过多种途径激活内皮型一氧化氮合酶（eNOS）、抑制活性氧（ROS）、抑制胆固醇晶体（CC）的产生进而调控TLR/NF-κB与NLRP3炎性小体通路,抑制TLR/NLRP3介导的炎症反应。同时,中医药中多种单味药有效成分与方剂可有效激活TRPV1/TRPA1或其下游途径,其对动脉粥样硬化中TLR/NLRP3通路可能具有显著调控作用。该文进一步对已探明的方剂及中药单味药有效成分调控AS中TLR/NLRP3通路的机制进行梳理,将TRPV1/TRPA1调控TLR/NLRP3途径的机制与中药方剂与单体成分进行对应。为治疗动脉粥样硬化的中药药物相互作用机制提供新启发...     

多胺在动脉粥样硬化发生发展过程中的变化

目的：探讨动脉粥样硬化的发病机制。方法：采用雄性日本鹌鹑喂饲高胆固醇膳食,造成动脉粥样硬化动物模型,动脉观察主动脉壁的中内膜的多胺变化。结果：血浆胆固醇和分类胆固醇提示高脂血症,结合病理切片,说明本实验动脉粥样硬化模型可靠。结论：1．多胺对动脉粥样硬化的发生起着重要作用;2．多胺在动脉粥样硬化发生（病理标准）前升高,后又依次降低的特殊现象。     

Variations of Concentration Characteristics of Rainfall Runoff Pollutants in Typical Urban Living Areas

Based on a typical residential area, this paper studies the characteristics of pollutant concentration changes in two rainfall runoffs and the first flush effect of rainfall. In rainfall runoff, the concentrations of seven pollutants (COD_Mn, TN, DTN, NH₃-N, TP, DTP, and PO₄³⁻) increased during the initial rainfall period and decreased in the later period. Rainfall causes the erosion of pollutants on the underlying surface so that water pollution begins when rainfall runoff occurs, and the pollution level drops over time. The seven pollutants all experience this first flush effect, of which, rainfall has the strongest scouring effect on NH₃-N produced by domestic sewage. The significant excess of pollutants in rainfall runoff should be considered by management departments. In addition, the existence of the first flush effect makes it possible in theory to partially intercept rainfall runoff to control water pollution, thereby reducing the cost of pollution control.