A facile fluorescence turn-on biosensor customized for monitoring of protein kinase activity based on carboxylic carbon nanoparticle–peptide complexes

In this study, we present a facile and low-cost approach for detecting protein kinase A (PKA) by assembling a purpose-designed carboxyfluorescein (FAM)-labelled peptide with carboxylic carbon nanoparticles (CNPs). Fluorescence of the FAM-labelled peptide gradually decreases to a low background signal as a result of the electron transfer from CNPs to FAM-labelled peptide via the peptide, which acts as a bridge. The reaction in the sensor in the presence of adenosine 5′-triphosphate and PKA phosphorylates the substrate peptide and disrupts the electrostatic repulsive force between the CNPs and the peptide, therefore altering the spectroscopic signal of the system. The change in fluorescence signal was directly proportional to the PKA concentration in the range 0–1.8 U/ml with a detection limit of 0.04 U/ml. These results suggest that PKA activity can be effectively measured using the developed PKA biosensor. Moreover, the fluorescence biosensor was successfully used in the investigation of PKA in spiked human embryonic kidney (HEK) 293 cells lysates, indicating its potential applications in protein kinase-related biochemical fundamental research.     

Nitrogen-doped carbon dots as a fluorescent probe for the highly sensitive detection of bilirubin and cell imaging

Nitrogen-doped carbon dots (NCDs) with bright blue fluorescence were constructed by a hydrothermal method using sucrose and l- proline as raw materials. The NCDs were characterized by transmitted electron microscopy, X-ray diffraction, Fourier-transform infrared spectrometry, X-ray photoelectron spectroscopy, and ultraviolet-visible absorption and fluorescence spectroscopy to investigate the morphology, elemental composition, and optical properties. The NCDs had good water solubility, high dispersibility with an average diameter of only 1.7 nm, and satisfactory optical properties with a fluorescence quantum yield of 23.4%. The NCDs were employed for the detection of bilirubin. A good linear response of the NCDs in the range 0.35–9.78 μM was obtained for bilirubin with a detection limit of 33 nM. The NCDs were also applied to the analysis of real samples, serum and urine, with a recovery of 95.34% to 104.66%. The low cytotoxicity and good biocompatibility of the NCDs were indicated by an MTT assay and cell imaging of HeLa cells. Compared with other detection systems, using NCDs for bilirubin detection was a facile and efficient method with good selectivity and sensitivity.     

Utilization of N,S-doped carbon dots as a fluorescent nanosensor for determination of cromolyn based on inner filter effect: application to aqueous humour

A simple and eco-friendly hydrothermal technique is used to prepare water soluble N- and S-co-doped carbon quantum dots probes (N,S-CQDs) from thiosemicarbazide and citric acid. Several characterization techniques were performed to ensure the successful synthesis of highly luminescent N,S-CQDs. The prepared probe exhibited analytical potential as an optical nanosensor for the spectrofluorimetric determination of cromolyn sodium (CRO) in its pharmaceutical dosage forms and aqueous humour. The emission intensity of the synthesized N,S-CQDs was measured at 411 nm after excitation at 345 nm. Addition of increasing concentrations of CRO to N,S-CQDs led to quenching of its fluorescence intensity. CRO was investigated within a wide concentration range 10.0–150.0 μM with a limit of detection of 2.0 μM and a limit of quantification of 6.0 μM. The quenching of fluorescent N,S-CQDs occurred through the inner filter effect (IFE). The developed spectrofluorimetric method was successfully optimized and validated according to the International Council of Harmonisation guidelines (ICH). The method greenness is proved through using both Eco-Scale and AGREE approaches.     

盐酸多奈哌齐联合综合康复训练对老年痴呆患者认知功能、事件相关电位及血清BDNF、IGF-1的影响

目的:探讨盐酸多奈哌齐联合综合康复训练对老年痴呆患者认知功能、事件相关电位及血清脑源性神经营养因子(BDNF)、胰岛素样生长因子1(IGF-1)的影响。方法:选取2017年3月~2020年3月期间来我院接受治疗的120例老年痴呆患者,根据随机数字表法分为对照组(n=60,盐酸多奈哌齐治疗)和联合组(n=60,盐酸多奈哌齐联合综合康复训练治疗)。对比两组疗效,治疗前、治疗3个月后的认知功能评分、事件相关电位、血清BDNF和IGF-1水平变化,以及治疗期间不良反应发生情况。结果:联合组的临床总有效率为91.67%,高于对照组的70.00%(P<0.05)。两组治疗3个月后日常生活能力评估量表(ADL)、临床痴呆量表(CDR)评分降低,且联合组低于对照组(P<0.05);简易精神状态量表(MMSE)评分升高,且联合组高于对照组(P<0.05)。两组治疗3个月后N1潜伏期、P2潜伏期均缩短,P3波幅均升高,且联合组治疗3个月后P2潜伏期短于对照组,P3波幅高于对照组(P<0.05);两组治疗3个月后N1潜伏期比较未见统计学差异(P>0.05)。联合组治疗3个月后BDNF水平高于对照组(P<0.05);两组治疗3个月后IGF-1水平比较未见统计学差异(P>0.05)。对照组与联合组不良反应发生率对比无统计学差异(P>0.05)。结论:老年痴呆患者在盐酸多奈哌齐基础上联合综合康复训练,认知功能可得到明显改善,同时还可调节患者事件相关电位及血清BDNF、IGF-1水平。     

Current capacity,bottlenecks, and future projections for offsetting habitat loss using Mitigation and Conservation banking in the United States

Habitat banking in its many iterations is an established and popular mechanism to deliver environmental offsets. The United States can look back at over 30 years of banking experience with the underlying framework and policies being consistently updated and improved. Given the increased demand in habitat banking, we provide insights into how bank area capacity is distributed across the United States for four different bank targets (wetlands, streams, multiple ecosystems, species) based on information extracted from the Regulatory In-lieu Fee and Bank Information Tracking System, as well as, estimating future capacities and area reserves through a predictive modeling approach based on data from the past 26 years. Future predictions indicate a decrease in available reserves for banks targeting wetlands or multiple ecosystems, with potential bottlenecks relating to large reserves being limited to the southeast and release schedules not catching up to the current and anticipated demand. Banks targeting species or streams are predicted to meet future demand, with species banks (conservation banks) following a different legislative and operational approach based on the listing of endangered species and pro-active approaches with anticipated future demand. Most current reserves for all four bank types are restricted to very few service areas with around one-third of all bank areas still awaiting release, limiting their availability on a broader scale. Strategic planning networks are necessary to meet future demand on a national scale and to identify areas suitable for banking or likely to experience future environmental or developmental stress.     

组织工程支架制备中超临界CO2技术的应用*

作为组织工程研究中三大要素之一,组织工程支架可为细胞的附着、迁移和增殖提供理想的环境。传统的组织工程支架制备方法,如粒子沥滤法、相分离法及静电纺丝法等在理论和技术上已较为成熟,但由于大多需要有机溶剂的参与,在制备过程中仍存在有机溶剂难以去除,以及支架孔洞难以控制、连通性较差等问题。超临界二氧化碳(supercritical carbon dioxide,SC-CO₂)密度近似液体,黏度和扩散系数近似气体,具有流动性强、溶解能力大、传热效率高等特殊的理化性质,与传统工艺相结合,可在绿色温和的反应体系中有效规避上述问题,在组织工程支架制备及药物负载方面具有广阔前景。     

纳布啡鞘内注射对糖尿病神经痛模型大鼠行为能力及背根神经节TRPV1表达的影响

摘要 目的：探讨纳布啡鞘内注射对糖尿病神经痛（diabetic neuropathic pain, DNP）模型大鼠行为能力及背根神经节瞬时受体电位V1（transient receptor potential V1,TRPV1）表达的影响。方法：将糖尿病神经痛模型大鼠（n=48）随机平方为三组-模型组、纳布啡1组与纳布啡2组,每组16只。纳布啡1组与纳布啡2组分别给予纳布啡鞘内注射0.5 μg/10 μL与1.0 μg/10 μL,模型组给予注射等剂量的0.9 %氯化钠溶液,每天1次。分别于治疗第7 d、第14 d,采用血糖仪测定与记录空腹血糖（fasting blood glucose,FBG）水平并进行机械痛阈检测;治疗第14 d、第28 d,采用酶联免疫法检测血清IL-6与TNF-α含量,采用免疫印迹法检测背根神经节TRPV1蛋白的相对表达。结果：模型组、纳布啡1组与纳布啡2组治疗第7 d、第14 d的空腹血糖水平都高于20.00 mmol/L,组内与组间对比差异不具有统计学意义（P>0.05）。纳布啡1组与纳布啡2组治疗第7 d、第14 d的机械痛阈高于模型组（P<0.05）,也高于治疗前（P<0.05）,纳布啡2组与纳布啡1组差异具有统计学意义（P<0.05）。纳布啡1组与纳布啡2组治疗第14 d、第28 d的血清白细胞介素（Interleukin,IL）-6与肿瘤坏死因子（Tumor necrosis factor,TNF）-α含量明显低于模型组（P<0.05）,纳布啡2组明显低于纳布啡1组（P<0.05）。纳布啡1组与纳布啡2组治疗第14 d、第28 d的背根神经节TRPV1相对表达水平明显低于模型组（P<0.05）,纳布啡2组明显低于纳布啡1组（P<0.05）。结论：纳布啡鞘内注射在糖尿病神经痛模型大鼠的应用能改善行为能力,抑制背根神经节TRPV1的表达,还可抑制血清IL-6与TNF-α的释放,从而发挥镇痛治疗效用。     

玉米秸秆还田对黑土微生物群落功能多样性的影响

为了解玉米秸秆还田对土壤微生物群落功能多样性的影响,采用ECO-Biolog微平板法,分析了以无玉米秸秆还田为对照(CK),6000 kg·hm^-2(S1)、9000 kg·hm^-2(S2)、12000 kg·hm^-2(S3)和15000 kg·hm^-2(S4)四个秸秆还田量的土壤微生物碳源代谢特征。结果表明:随着玉米秸秆还田量的增加,土壤微生物平均颜色变化率(AWCD)也随之增加,24~96 h的AWCD值变化迅速,96 h后进入平稳期,S4处理AWCD值始终大于其他处理;秸秆还田对Shannon指数与Simpson指数没有显著影响(P>0.05),但与CK相比,S4处理McIntosh指数显著增加了57.5%(P<0.05);主成分分析结果显示,秸秆还田影响着土壤微生物群落碳代谢能力,S1、CK、S2和S3处理在PC1和PC2上出现显著的分异,糖类、多聚物类、羧酸类碳源是研究区域内土壤微生物利用的主要碳源。因此,在东北黑土区增加玉米秸秆还田量能够提高土壤微生物对碳源的利用能力,提升黑土肥力。     

基于MaxEnt模型预测气候变化下杉木在中国的潜在地理分布

为研究杉木在中国的分布特征及其对气候变化的响应模式,本研究基于现有分布记录,应用最大熵(MaxEnt)模型和地理信息系统方法,结合气候、地形等环境要素,预测杉木在当前和未来气候变化下的潜在适生区。结果表明: 影响杉木分布的最主要因素是年平均降水量,在当前气候下,杉木适生区合计面积328万km²,占全国陆地总面积的34.5%,低、中和高适生区分别占18.3%、29.7%与52.0%。在未来气候情景下,杉木生长的适宜性在我国总体上呈上升趋势,适生区面积随气候变化增大,且明显向北扩张,南方湿润亚热带地区形成集中连片高适生区。模型经受试者工作特征曲线检验,训练集平均受试者工作特征曲线下面积为0.91,可信度高。     

放牧对黄土高原退耕草地土壤有机碳储量的影响

放牧是影响草地土壤碳固存的重要因素。本研究选取黄土高原水蚀风蚀交错区西部、中部、东部地区及水蚀区,以各区20年以上退耕封禁地为对照,分析3个放牧强度下(羊粪球密度分别为0～10、10～20、>20 ind·m^-2)退耕草地0～20 cm土层土壤有机碳储量的分布特征,研究放牧及其强度对退耕草地土壤固碳效应的影响。结果表明: 放牧对交错区西部0～20 cm、东部0～10 cm,水蚀区0～5 cm土层土壤有机碳储量有显著影响,对交错区中部各土层均无显著影响;羊粪球密度0～10、＞20 ind·m^-2强度的放牧使交错区西部0～20 cm土层土壤有机碳储量显著降低了34.8%～50.9%,而在其他3个区域,放牧对有机碳储量的影响较退耕封禁地差异不显著。在交错区东部,放牧强度是影响退耕草地土壤有机碳储量的主要因素,而其他3个区域有机碳储量主要受土壤理化性质和(或)枯落物生物量的影响。羊粪球密度10～20 ind·m^-2强度的放牧对各区域退耕草地0～20 cm土层土壤有机碳储量无显著影响。