泪液蛋白质组学分析在眼部疾病中的应用及研究进展

泪液由泪腺及副泪腺分泌,主要通过眼睑和瞬目运动构成泪膜的水样层。泪液的蛋白质组学等研究是当前面临的难点也是热点,其主要目的是研究眼部疾病甚至部分全身疾病的代表其特异性和高敏感性的易检测出的生物标志物,以提高临床疾病的诊断和监测,还可通过识别疾病的靶点促进个性化治疗以及改善预后,并可以设计有针对性的后续实验了解疾病分子机制。在眼部疾病和部分全身疾病中或比血液、尿液、脑脊液等现有常规体液检查更有应用前景的标本检测。目前大量研究表明很多疾病在泪液中有成分的变化,本文就应用蛋白质组学的方法探讨眼部疾病在泪液中的蛋白质成分变化进行综述。

Construction and expression of retroviral vector pLEGFP-N1-TERT in preparation of seed cells for skin tissue engineering

Objective:To construct the retroviral vector pLEGFP-N1-telomerase reverse transcriptase(TERT)and to investigate the expression of TERT in neonatal mouse bypodermal cells.Methods:The polymerase chain reaction(PCR)-amplified TERT gene was inserted into plasmid pLEGFPN1.The positive clone was identified by restriction enzyme digestion and sequencing,then was transfected into packaging cells to produce retrovirus particles.Neonatal mouse hypodermal cells were infected with the virus to generate a stable cell line.The TERT mRNA expression level,telomerase activity,and enhanced green fluorescent protein(EGFP)expression level were analyzed.Results:Retroviral vector pLEGFP-N1-TERT was constructed successfully,and a stable cell line of neonatal mouse hypodermal cells expressing EGFP was established.Western blot and immunohistochemical assay showed that the expression level of TERT was significantly elevated in the neonatal mouse hypodermal cells.Conclusions:A high titer of retrovirus pLEGFP-N1-TERT mediates high-level expression of the exogenous TERT gene in the neonatal mouse hypodermal cells.This protocol has potential applications for skin tissue engineering and cell transplantation therapy.

Hybridization of iron phthalocyanine and MoS_(2)for high-efficiency and durable oxygen reduction reaction

Hybrid catalysts based on iron phthalocyanine(FePc)have raised much attention due to their promising applications in electrocatalytic oxygen reduction reaction(ORR).Various hybridization strategies have been developed for improving their activity and durability.However,the influence of different hybridization strategies on their catalytic performance remains unclear.In this study,Fe Pc was effectively distributed on molybdenum disulfide(MoS_(2))forming Fe Pc-based hybrid catalysts,namely Fe Pc-MoS_(2),Fe Pc*-MoS_(2),and Fe Pc-Py-MoS_(2),respectively,to disclose the related influence.Through direct hybridization,the stacked and highly dispersed Fe Pc on MoS_(2)resulted in Fe Pc-MoS_(2),and Fe Pc*-MoS_(2),respectively,in which the substrate and Fe Pc are mainly bound through van der Waals interactions.Through covalent hybridization strategy using pyridyl(Py)as a linker,Fe Pc-Py-MoS_(2)hybrid catalyst was prepared.Experimental and theoretical results disclosed that the linker hybridization of Fe Pc and MoS_(2)facilitated the exposure of Fe-N4 sites,maintained the intrinsic activity of Fe Pc by forming a more dispersed phase and increased the durability via Fe-N bonding,rendering the Fe Pc-Py-MoS_(2)an excellent ORR hybrid catalyst.Compared with van der Waals hybridized Fe Pc-MoS_(2)and Fe Pc*-MoS_(2)in alkaline media,the linker hybridized Fe Pc-Py-MoS_(2)showed an obviously enhanced ORR activity with a half-wave potential(E_(1/2))of 0.88 V vs RHE and an ultralow Tafel slope of 26 m V dec-1.Besides,the Fe Pc-Py-MoS_(2)exhibited a negligible decay of E_(1/2) after 50,000 CV cycles for ORR,showing its superior durability.This work gives us more insight into the influence of different hybrid strategies on Fe Pc catalysts and provides further guidance for the development of highly efficient and durable ORR catalysts.

Highly selective photocatalytic oxidation of methane to methyl hydroperoxide

Partial oxidation of methane into primary oxidation products with high value remains a challenge.In this work,photocatalytic oxidation of methane(CH_(4))with high methyl hydroperoxide(CH_(3)OOH)selectivity is achieved using pure titanium oxide(TiO_(2))without any cocatalyst at room temperature and atmospheric pressure.The CH_(3)OOH production rate can reach up to 2050±88μmol·g^(-1)·h^(-1) at pH≈7.0 with 100%selectivity in the liquid product.The stable reaction cycle can reach more than 30 times.This low-cost system achieves superior CH_(4) conversion activity and selectivity compared with similar work.The energy of hydrogen peroxide(H_(2)O_(2))to adsorbed hydroperoxyl radical(^(*)OOH)has a significantly lower reaction energy than conversion to adsorbed hydroxyl radical(*OH)on the(210)surface of the TiO_(2).The^(*)OOH preferentially combines with methyl radical(·CH_(3))to form the most energetically favorable CH_(3)OOH.The mild oxidative environment of this system prevents the reduction of CH_(3)OOH to CH_(3)OH or over-oxidation of CH_(4),which ensures the final CH_(3)OOH with high selectivity and stability.This work provided a low-cost but highly efficient method to achieve partial oxidation with superior selectivity,i.e.,to convert CH_(4) into high-value chemicals.

Limitation standard of toxic aconitines in Aconitum proprietary Chinese medicines using on-line extraction electrospray ionization mass spectrometry

Development of rapid analytical methods and establishment of toxic component limitation standards are of great importance in quality control of traditional Chinese medicine. Herein, an online extraction electrospray ionization mass spectrometry(oEESI-MS) coupled with a novel whole process integral quantification strategy was developed and applied to direct determination of nine key aconitine-type alkaloids in 20 Aconitum proprietary Chinese medicines(APCMs). Multi-type dosage forms(e.g., tablets, capsules, pills, granules, and liquid preparation) of APCM could be determined directly with excellent versatility. The strategy has the characteristics of high throughput, good tolerance of matrix interference, small amount of sample(~0.5 mg) and reagent(~240 μL) consumption, and short analysis time for single sample(<15 min). The results were proved to be credible by high performance liquid chromatographyàmass spectrometry(LC-MS) and electrospray ionization mass spectrometry, respectively. Moreover, the limitation standard for the toxic aconitines in 20 APCMs was established based on the holistic weight toxicity(HWT) evaluation and the Chinese Pharmacopoeia severally, and turned out that HWT-based toxicity evaluation results were closer to the real clinical applications. Hence,a more accurate and reliable APCM toxicity limitation was established and expected to play an important guiding role in clinics. The current study extended the power of ambient MS as a method for the direct quantification of molecules in complex samples, which is commonly required in pharmaceutical analysis,food safety control, public security, and many other disciplines.

Nonlinear characterization and performance optimization for broadband bistable energy harvester

As two crucial indicators of bistable energy harvesting performance,band width and power amplitude are simultaneously investigated for obtaining the synergy effect.Toward this goal,a nonlinear electromechanical-coupled distributed-parameter model of the bistable piezoelectric energy harvester is established.Based on the electromechanical decoupled method,approximate higher-order analytical solutions of the beam displacement,harvested power and effective bandwidth are derived.The cubic-function discriminant of the analytical solution is introduced to determine the nonlinear excitation-frequency boundaries of multiple solutions and power peak.The stability of the multiple solutions is analyzed through Jacobi matrix of the modulation equation.Superharmonic resonance is notified.Upward and downward sweep experiments and numerical solutions of time history curves,phase portraits and power spectra confirm the analytical findings.To realize optimized broadband energy harvesting,the parametric study on the coefficients of the linear and cubic elastic external forces with the corresponding optimal load resistance is performed.For the nonlinear hardening case,more positive linear coefficient is preferred.For the nonlinear softening case,the cubic coefficient slightly larger than its optimal value is recommended at each given linear coefficient.By tuning the load resistance and linear and cubic coefficients of the external force,broadband bistable energy harvesting with optimized power is realized.

Anisotropic iron-doping patterns in two-dimensional cobalt oxide nanoislands on Au(111)

An in teg rated approach combi ning den sity functional theory(DFT)calculati ons and atomic resol utio n sea nning tun neling microscopy(STM)is used to study well-defined iron-doped cobalt oxide nanoislands supported on Au(111).The focus is on the structure and distribution of Fe dopants within these nanoislands of CoO as a function of Fe to Co ratio.The DFT and STM results agree strongly and complement each other to allow for a more complete un dersta nding of the dopa nt structure trends on the nano scale.Using Fe as a marker,we first find that the stacki ng seque nee of the moire structure of the host cobalt oxide nano islands can be ide ntified un ambiguously through a combinati on of DFT and STM.Using the distinct con trast of the embedded Fe dopa nt atoms as observed with atom-resolved STM,we find correlatio ns betwee n Fe dopa nt positi on and the CoO/Au(111)moire patter n at varyi ng Fe dopa nt den sities.Formatio n of Fe-dopa nt clusters within the nano islands is investigated in detail through DFT and found to agree with the dopant patter ns observed in STM.We find that the structural effects of Fe dopants throughout the nanoislands with the basal planes and the two types of edges—the oxygen and metal edges—have different nature.Both DFT calculations and STM images show a strong preferenee for Fe dopants to be located directly on or near the oxygen edge of the nanoislands as opposed to being directly on or n ear the metal edge.Take n together,our results illustrate that Fe dopa nt incorporati on and distributi on within CoO nanoislands are highly an isotropic and gover ned by both the moire structure of the basal planes as well as nano-size effects present at the under-coordinated edges of different local geometry and chemistries.

DES-Fe3O4 composite for rapid extraction of residual plant growth regulators in edible vegetable oil

A rapid DES-Fe3O4 microextraction coupled with liquid chromatography method was ingeniously developed for simultaneous control of five typical plant growth regulators in a planting cycle including indole-3-acetic acid, abscisic acid, thidiazuron, 1-naphthylacetic acid and forchlorfenuron. The experiments were carried out with one type of carboxylic acid-assisted deep eutectic solvent (nChCl:nacetic acid=1:3) as extractant, coupled with ultrasonic wave extraction for 30 min at bath temperature at 50 ℃. Under the optimum conditions, good extraction performances of linearities for five plant growth regulators were achieved separately in the range of 0.10–50 mg/L of indole-3-acetic acid, 0.06–50 mg/L of abscisic acid, 0.20–50 mg/L of thidiazuron, 0.50–50 mg/L of 1-naphthylacetic acid and 0.06–50 mg/L of forchlorfenuron and recoveries were ranged from 70.4%to 102.2%. Meanwhile, Fe3O4 was preferentially combined with DES in the oil matrix, the advantage of which was pointed to rapid and efficient separation of targeted plant growth regulators from complicated oil matrix. The proposed approach achieved satisfied results and met the monitoring requirements of plant growth regulators control in edible vegetable oil samples.

球几何区域上Helmholtz传输特征值问题有效的谱Galerkin逼近

本文提出球几何区域上Helmholtz传输特征值问题基于Legendre-Galerkin逼近的一种有效的谱方法.首先,通过球坐标变换和球调和函数展开,将原问题化为一系列等价的一维广义特征值问题.然后,针对每个一维广义特征值问题建立相应的弱形式和离散格式,并利用紧算子的谱理论证明特征值和特征函数的误差估计.特别地,对于实心的球形区域,需要克服由球坐标变换引入的极点奇性这一困难.因此,本文推导了相应的极条件,并根据极条件引入带权的Sobolev空间,从而建立了每个一维广义特征值问题相应的弱形式和离散格式.最后给出一些数值例子,数值结果表明算法的有效性和理论结果的正确性.

A novel aptamer-based histochemistry assay for specific diagnosis of clinical breast cancer tissues

Pathological detection using immunohistochemistry(IHC)has become an indispensable process in the diagnosis confirmation of various cancers.However,the production of monoclonal antibodies is always very complex,expensive and time-consuming,and the batch differences are significant due to the corporeity and health statuses of animals may be different.In this work,an aptamer-based histochemistry(aptahistochemistry)assay was developed using a DNA aptamer for specific diagnosis of clinical breast cancer tissue sections.This aptahistochemistry assay can specifically distinguish Luminal A breast cancer molecular subtype from Luminal B(HER2+),HER2-enriched,and triple-negative breast cancer molecular subtypes,as well as para-carci noma tissue,mastitis tissue and normal breast tissue.The accuracy of this aptahistochemistry assay for the diagnosis of Luminal A breast cancer was as high as 80%,which showed a great potential for clinical pathological diagnosis applications.