聚对氯苯胺修饰电极的制作及应用研究

通过对聚对氯苯胺膜修饰电极的制作和实验特性的研究,发现聚对氯苯胺膜表面均匀、光滑、牢固,在pH3～11范围内均有响应,测定的重现性好。电极使用长久且稳定性高。该电极和参比电极可直接插入家兔等活体内测定血乳酸的变化情况,对生命科学的活体检测和诊断有其实用意义。     

基于Arduino的智能送药小车

智能单片机小车由于其智能、方便、快捷,被广泛应用于各行业.本研究的智能送药小车包括了Ardunio mega2560单片机、OpenMv4智能识别装置、寻迹装置、L298N模块、433M无线模块等装置.研究结果表明:智能送药小车通过识别地面安置路线以及相应的病房号,可以自动躲避障碍,自动沿设定的路线行驶,既可以实现单车...     

稻田转为菜地初始阶段温室气体排放特征

【目的】近年来,随着我国社会经济的快速发展和人们生活水平的提高及膳食结构的改善,越来越多的稻田被转为蔬菜种植,影响了土壤碳氮转化过程及其引起的温室气体排放。因此有必要探究稻田转为蔬菜种植,特别是该土地利用方式转变初始阶段的温室气体（CH₄和N₂O）排放特征及其关键影响因素。【方法】试验选取了长期种植水稻的双季稻田,将其中一部分转为蔬菜种植,另一部分继续种植水稻,每个处理设置了3个重复,按照当地常规模式进行管理。采用静态暗箱—气相色谱法连续3年进行田间原位观测,比较分析稻田和由稻田转变的菜地CH₄和N₂O排放特征及其年际变化差异,明确稻田转为菜地初始阶段CH₄和N₂O排放的关键影响因素。【结果】稻田是重要的CH₄排放源,其第一年的排放强度（183.91 kg CH₄-C·hm^-2?a^-1）明显低于后续两年（241.56—371.50 kg CH₄-C·hm^-2?a^-1）,这主要归功于后两年降雨量的增加引起了土壤水分含量的升高。稻田转为菜地显著减少了CH₄排放,减少量相当于稻田CH₄年累积排放量的83%—100%。菜地第一年的CH₄累积排放量（31.22 kg CH₄-C·hm^-2）显著高于第二年（0.45 kg CH₄-C·hm^-2）和第三年（0.89 kg CH₄-C·hm^-2）,表明稻田转菜地对CH₄排放的影响具有时间滞后效应。稻田是弱的N₂O排放源（1.35—3.49 kg N₂O-N·hm^-2?a^-1）,其转为菜地显著增强了N₂O排放。菜地第一年的N₂O累积排放量（95.12 kg N₂O-N·hm^-2）显著高于第二年（38.28 kg N₂O-N?hm^-2）和第三年（40.07 kg N₂O-N·hm^-2）。菜地土壤异养呼吸对N₂O排放的影响在第一年明显高于第二、三年,表明稻田转为蔬菜种植的第一年,有机质矿化对N₂O排放有重要贡献。在100年尺度CO₂当量下,稻田转为蔬菜种植第一和第二年的综合增温潜势（GWP）相对于稻田分别显著增加了390%和98%,主要是由于增加的N₂O增温潜势超过了减少的CH₄增温潜势。但是,稻田转为菜地的第三年,菜地的GWP（(16.72±3.25) Mg CO₂-eq·hm^-2）与稻田（(14.84±1.39) Mg CO₂-eq·hm^-2）相比无显著差异,主要是由于减少的CH₄ 增温潜势完全抵消了增加的N₂O增温潜势。这些研究结果表明稻田转菜地对GWP的影响主要集中在该土地利用方式转变的第一年。【结论】稻田转为菜地显著减少了CH₄排放,增加了N₂O排放,增强了菜地第一和第二年的综合增温潜势。有机质矿化过程对新转菜地第一年较高的N₂O排放有重要贡献。这些研究结果表明了评价土地利用方式转变初始阶段温室气体排放特征的重要性,便于及时采取有效管理措施缓解温室气体排放,实现环境友好型农业可持续生产。     

辛基酚对家蚕（Bombyx mori L.）生长发育的影响

为探讨辛基酚对鳞翅目模式昆虫家蚕生长发育的影响,使用添加0.01～0.08 g·kg~(-1)辛基酚(4-tert-octylphenol,4-t-OP)的人工饲料饲养家蚕(Bombyx mori L.),研究了4-t-OP对家蚕幼虫和蛹的生长发育速度、生长量、发育整齐度和生命力等影响。结果表明:连续取食0.01 g·kg~(-1)以上浓度4-t-OP的饲料,家蚕幼虫和蛹的生长发育速度(幼虫期和蛹期发育经过时间)、生长量(幼虫期体质量和蛹期茧层量)、生命力(3龄起蚕绝食生命时数、4龄起蚕结茧率、蚕茧的死笼率和羽化率)等都明显下降,抑制眠蚕体质量。4-t-OP对家蚕幼虫生长发育后期、化蛹及羽化等发育变态期的影响更加明显,其在家蚕体内可能有累积效应。添食0.08 g·kg~(-1)4-t-OP的家蚕不能完成生活史。研究表明,4-t-OP对家蚕生长发育毒性影响有显著的浓度效应和时间效应。     

胡椒4-香豆酸：辅酶A连接酶Pn4CL基因的克隆及表达分析

根据胡椒4-香豆酸：辅酶A连接酶（4-coumarate:coenzyme A ligase, 4CL）基因的部分序列设计引物,运用RACE方法获得其家族成员的1个全长cDNA,命名为Pn4cl,长度2130 bp,开放阅读框1638 bp,编码545个氨基酸。预测Pn4CL分子量为59.57 kDa,理论等电点为5.70。该基因含有AMP-binding（AMP-binding enzyme）、CaiC[Acyl-CoA synthetase (AMP-forming) /AMP-acid ligaseⅡ]、PLN02246、AFD-class I等结合域,具有植物4CL所共有的保守结构域。系统进化分析表明,Pn4CL与北细辛的同源性最高,同时与木兰分支类植物的4CL聚类在一起,与菊分支的进化距离较近,与蔷薇分支的进化距离较远。亚细胞定位表明,该蛋白定位在细胞膜上。Real-time RT-PCR结果表明,该基因受外援激素SA和MeJA诱导表达,同时接种辣椒疫霉菌后,Pn4CL基因的表达量在抗/感2种胡椒中均出现先增加后减少的现象,并且在抗病种质中表达量较高。研究结果为Pn4CL的功能研究提供了理论依据。     

Verrucosamide,a Cytotoxic 1,4-Thiazepane-Containing Thiodepsipeptide from a Marine-Derived Actinomycete

A new cytotoxic thiodepsipeptide, verrucosamide (1), was isolated along with the known, related cyclic peptide thiocoraline, from the extract of a marine-derived actinomycete, a Verrucosispora sp., our strain CNX-026. The new peptide, which is composed of two rare seven-membered 1,4-thiazepane rings, was elucidated by a combination of spectral methods and the absolute configuration was determined by a single X-ray diffraction study. Verrucosamide (1) showed moderate cytotoxicity and selectivity in the NCI 60 cell line bioassay. The most susceptible cell lines were MDA-MB-468 breast carcinoma with an LD₅₀ of 1.26 µM, and COLO 205 colon adenocarcinoma with an LD₅₀ of 1.4 µM. Also isolated along with verrucosamide were three small 3-hydroxy(alkoxy)-quinaldic acid derivatives that appear to be products of the same biosynthetic pathway.     

Sulodexide protects against hyperglycemia-induced retinal vascular injury via suppressing NOX4/NLRP3 pathway

AIM To investigate the effect of sulodexide (SDX) on high glucose-induced damage in retinal microvascular endothelial cells. METHODS (1) High-fat diet combined with intraperitoneal injection of streptozocin were used to induce type 2 diabetes mellitus (DM) followed by injection of saline or SDX in C57BL/6J male mice. Retinal microvascular leakage and density, and the protein levels of NLRP3 inflammasome-related proteins, zonula occludens-1 (ZO-1) and NADPH oxidase 4 (NOX4) were measured. (2) Human retinal microvascular endothelial cells (HRMECs) were treated with normal glucose or high glucose with or without SDX, and were further transfected with siRNA to knock down NOX4, or infected by adenovirus to over-express NOX4. The protein levels of ZO-1, VE-cadherin (VE-Cad), NOX4 and NLRP3 inflammasome-related proteins as well as the level of reactive oxygen species (ROS) were detected. RESULTS Treatment with SDX increased the protein level of ZO-1, attenuated retinal leakage and NLRP3 inflammasome activation, and enhanced the density of microvasculature and the number of ganglion cells in diabetic retinas. The protein levels of ZO-1 and VE-Cad were decreased, while the levels of NOX4, NLRP3 inflammasome-related proteins and ROS generation were increased in high glucose-treated HRMECs. Silencing of NOX4 inhibited high glucose-induced increases in NLRP3 inflammasome and ROS generation, and decreases in the protein levels of ZO-1 and VE-Cad. Over-expression of NOX4 significantly increased the levels of NLRP3 inflammasome-related proteins and ROS generation in HRMECs, and reduced the protein levels of ZO-1 and VE-Cad. Treatment with SDX partly reversed NOX4 over-expression-induced changes. CONCLUSION SDX alleviates hyperglycemia-induced retinal microvascular endothelial injury via inhibiting NOX4/ROS/NLRP3 pathways.     

Roles of protein phosphatase 4 in down-regulation of eNOS Ser633 phosphorylation induced by palmitic acid in human umbilical vein endotheli?al cells

AIMTo investigate the roles of protein phosphatase 4 (PP4) in down-regulation of endothelial nitric oxide synthase (eNOS) Ser633 phosphorylation induced by palmitic acid (PA). METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with PA at 25 μmol/L, 50 μmol/L, 100 μmol/L and 200μmol/L for 36 h, or treated with PA at 100 μmol/L for 12 h, 24 h, 36 h and 48 h. Protein phosphatase 2A (PP2A) family inhibitor fostriecin (FST, 20 nmol/L) or okadaic acid (OA, 5 nmol/L) was selected to pretreat the HUVECs for 30 min. Protein phosphatase 4 catalytic subunit (PP4c) siRNA or protein phosphatase 2A catalytic subunit (PP2Ac) siRNA was transfected into the HUVECs. The protein expression levels of of eNOS, PP4c and PP2Ac, as well as the level of eNOS Ser633 phosphorylation, were detected by Western blot. The intracellular nitric oxide (NO) content was measured by DAF-FM DA. RESULTS(1) Compared with control group, the levels of eNOS Ser633 phosphorylation were decreased in PA groups in which the HUVECs were treated with 25 μmol/L, 50 μmol/L, 100 μmol/L and 200 μmol/L PA for 36 h (P<0.05) and 100 μmol/L PA for 24 h, 36 h and 48 h (P<0.05). No significant difference in the level of total eNOS protein expression among all the groups was observed. (2) Compared with control group, both FST and OA pretreatment reversed the reduction of eNOS Ser633 phosphorylation (P<0.05) and the decrease in intracellular NO content (P<0.05) induced by PA. No significant difference in the level of total eNOS protein expression among all the groups was observed. (3) Compared with si-Control group, the PP4c protein expression was significantly reduced (P<0.05), while the level of eNOS Ser633 phosphorylation was significantly increased in si-PP4c group (P<0.05). Although the levels of PP2Ac protein expression declined significantly (P<0.05), the level of eNOS Ser633 phosphorylation remained unchanged in si-PP2Ac group. No significant differencein the level of total eNOS protein expression among all the groups was found. CONCLUSION PA significantly reduces the level of eNOS Ser633 phosphorylation and the content of NO in the HUVECs, which may be due to PA inducing the activation of the PP2A family member PP4 rather than PP2A.     

Role of IP3R in LH-EGFR-induced mouse oocyte meiotic resumption

AIM: To explore the effect of inositol 1, 4, 5-trisposphate receptor (IP₃R) in luteinizing hormone-epidermal growth factor receptor (LH-EGFR)-induced oocyte meiotic resumption. METHODS: Models of mouse cumulus-oocyte complexs (COCs) culture and follicle culture in vitro were generated to study the effects of 2-aminoethyl diphenyl borate (2-APB) and heparin (IP₃R specific inhibitors) on LH/EGF-induced oocyte meiotic resumption and EGF-induced cumulus cell expansion. Real-time PCR was used to detect the mRNA expression of cumulus expansion-related factors. The changes of the intracellular calcium level were monitored using Fluo 3-AM, and the cGMP level was measured by ELISA. RESULTS: The inhibitors of IP₃R, 2-APB and heparin, dramatically reversed EGF-induced oocyte maturation (P<0.05) and decreased cGMP levels in COCs (P<0.05). In addition, 2-APB and heparin reversed EGF-induced cumulus expansion, and significantly inhibited EGF-induced cumulus expansion-related factor expression (P<0.05). The activation of IP₃R increased intracellular calcium level, and the study found that 2-APB and heparin dramatically reversed EGF-induced elevation of calcium level in cumulus cells (P<0.05). Follicular culture in vitro showed that 2-APB and heparin significantly reversed the LH-induced oocyte maturation (P<0.05). CONCLUSION: LH-EGFR signaling pathway increases calcium level in cumulus cells through IP₃R, resulting in meiotic resumption.