A Convenient Fluorescent-labeled Assay for in vitro Measurement of DNA Mismatch Repair Activity

Objective The assay of DNA mismatch repair （MMR） activity can be used as a biomarker for environmental condition detection and human disease diagnosis. Radioactive 32p-endlabeled DNA containing mismatch is extensively used as the substrate for MMR activity analyses. The aim of the present study is to develop a simple non-radioactive, but equally specific and sensitive method for the MMR activity assay. Methods A fluorescent label was chosen to replace the radioactive isotope label. Sensitive evaluation of the fluorescent label was carried out for the first time, and then the fluorescent label was compared with the isotope label in the MMR activity and DNA binding assays. Result LOD （limit of detection） of the fluorescent label was about 0.1 fmol and the relative signal strength displayed a pretty good linear relationship. Moreover, the fluorescent label method has equivalent sensitivity and performance as compared with the classical radioactive method in experiments. Conclusion In light of the sensitivity, reproducibility, safety, rapidity and long lifespan of the fluorescent label, this improved method can be applied to evaluation of biologic and toxic effects of environmental pollutants on man and other forms of life.

Preparation and Performance of Fluorescein Isothiocyanate⁃Labeled Fluorescent Starch and Polyvinyl Alcohol for Warp Sizing

In order to solve the problems of low accuracy and poor variety adaptability of the current measurement method of the permeability and coating property of sizing paste in textile industry, taking starch and polyvinyl alcohol(PVA) as the representatives of the most commonly used textile sizes, various concentrations of fluorescent molecules fluorescein isothiocyanate(FITC) were used to label starch and PVA to prepare fluorescein(F)-starch and F-PVA fluorescent sizes with different degrees of labeling(DLs) for the first time, respectively. Then the starch and PVA derivatives were employed to size pure cotton warp yarns. After preparing the sections of the sized yarns, the permeability and coating percentage of starch and PVA paste to the yarns were calculated by using a fluorescence microscope and the Photoshop software, respectively. The results demonstrate that F-starch and F-PVA with appropriate DL of 0.791% and 0.161%, respectively, exhibit good fluorescence property and similar sizing performance to the sizing performance of unlabeled starch and PVA. It is considered that fluorescence labeling of sizing agents with fluorescein units can provide an innovative way for the accurate determination of the permeability and coating property of sizing paste to warp yarns.

Construction of bicistronic green fluorescent protein labeled pSELECT GFPzeo human bone morphogenetic protein 2 eukaryotic expression vector

Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector.Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by using polymerase

The LB Films of Dansyl Chloride Labeled Octadecylamine and Its Fluorescence Lifetime

Octadecylamine was derivatized with dansyl chloride (5-dimethylaminonaphthalene-1-sulfonyl chloride) In order to simplify and understand the LB films of fluorescent probe labeling proteins. its monolayer and multilayers in the absence and presence of stearic acid were deposited by LB technique. Fluorescence spectra and lifetimes of the fluorescent products were studied to elucidate the microenvironment of molecules in the LB films.

RGDC Peptide Modified Quantum Dots Labelling and Imaging of Tumor Cells

The labelling and imaging of tumor cells were investigated via arginine-glycine-aspartic acidcysteine（RGDC） peptide-labelled quantum dots（QDs）. The results show that RGDC modified QDs can label SMMC-7721 tumor cells and adhere to cellular membrane. In constrast, the unmodified QDs are mainly dispersed around the cell. We also found that the RGDC-QDs can penetrate into the cell at 2 h of incubation. After 6 h of incubation, RGDC-QDs can accumulate in a unique intracellular region.

Visualization of integrin molecules by fluorescence imaging and techniques

Integrin molecules are transmembraneαβheterodimers involved in cell adhesion,trafficking,and signaling.Upon activation,integrins undergo dynamic conformational changes that regulate their affinity to ligands.The physiological functions and activation mechanisms of integrins have been heavily discussed in previous studies and reviews,but the fluorescence imaging techniques-which are powerful tools for biological studies-have not.Here we review the fluorescence labeling methods,imaging techniques,as well as Förster resonance energy transfer assays used to study integrin expression,localization,activation,and functions.

利用定量单病毒示踪技术探究SARS-CoV-2与细胞膜融合动态过程

Viral envelope fusion with the host plasma membrane(PM)for genome release is a hallmark step in the life cycle of many enveloped viruses.This process is regulated by a complex network of biomolecules on the PM,but robust tools to precisely elucidate the dynamic mechanisms of virus-PM fusion events are still lacking.Here,we developed a quantitative single-virus tracking approach based on highly efficient dual-color labelling of viruses and batch trajectory analysis to achieve the spatiotemporal quantification of fusion events.This approach allows us to comprehensively analyze the membrane fusion mechanism utilized by pseudotyped severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)at the singlevirus level and precisely elucidate how the relevant biomolecules synergistically regulate the fusion process.Our results revealed that SARS-CoV-2 may promote the formation of supersaturated clusters of cholesterol to facilitate the initiation of the membrane fusion process and accelerate the viral genome release.

Shaping Small Bioactive Molecules to Untangle Their Biological Function： A Focus on Fluorescent Plant Hormones

Modern biology overlaps with chemistry in explaining the structure and function of all cellular processes at the molecular level. Plant hormone research is perfectly located at the interface between these two disciplines, taking advantage of synthetic and computational chemistry as a tool to decipher the complex biological mechanisms regulating the action of plant hormones. These small signaling molecules regulate a wide range of developmental processes, adapting plant growth to ever changing environmental conditions. The synthesis of small bioactive molecules mimicking the activity of endogenous hormones allows us to unveil many molec- ular features of their functioning, giving rise to a new field, plant chemical biology. In this framework, fluores- cence labeling of plant hormones is emerging as a successful strategy to track the fate of these challenging molecules inside living organisms. Thanks to the increasing availability of new fluorescent probes as well as advanced and innovative imaging technologies, we are now in a position to investigate many of the dynamic mechanisms through which plant hormones exert their action. Such a deep and detailed comprehension is mandatory for the development of new green technologies for practical applications. In this review, we sum- marize the results obtained so far concerning the fluorescent labeling of plant hormones, highlighting the basic steps leading to the design and synthesis of these compelling molecular tools and their applications.

Virtual Fluorescence Translation for Biological Tissue by Conditional Generative Adversarial Network

Fluorescence labeling and imaging provide an opportunity to observe the structure of biological tissues,playing a crucial role in the field of histopathology.However,when labeling and imaging biological tissues,there are still some challenges,e.g.,time-consuming tissue preparation steps,expensive reagents,and signal bias due to photobleaching.To overcome these limitations,we present a deep-learning-based method for fluorescence translation of tissue sections,which is achieved by conditional generative adversarial network(cGAN).Experimental results from mouse kidney tissues demonstrate that the proposed method can predict the other types of fluorescence images from one raw fluorescence image,and implement the virtual multi-label fluorescent staining by merging the generated different fluorescence images as well.Moreover,this proposed method can also effectively reduce the time-consuming and laborious preparation in imaging processes,and further saves the cost and time.

Determination of genetic relationships between evergreen azalea cultivars in China using AFLP markers

Evergreen azaleas are among the most important ornamental shrubs in China.Today,there are probably over 300 cultivars preserved in different nurseries,but with little information available on the cultivar itself or relationships between cultivars.Amplified fragment length polymorphism(AFLP) markers were employed to determine the genetic relationships between evergreen azalea cultivars in China.One hundred and thirty genotypes collected from gardens and nurseries,including cultivars classified in the groups East,West,Hairy,and Summer,unknown cultivars,and close species,were analyzed using three primer pairs.A total of 408 polymorphic fragments were generated by AFLP reactions with an average of 136 fragments per primer pair.The average values of expected heterozygosity and Shannon's information index were 0.3395 and 0.5153,respectively.Genetic similarities were generated based on Dice coefficients,used to construct a neighbor joining tree,and bootstrapped for 100 replicates in Treecon V1.3b.Principal coordinate analysis(PCO) was performed based on Dice distances using NTSYS-pc software.The AFLP technique was useful for analyzing genetic diversity in evergreen azaleas.Cluster analysis revealed that cultivars in the West and Summer groups were quite distinct from other groups in the four-group classification system and that the East and Hairy groups should be redefined.

Blue light-induced rare-earth free phosphors for the highly sensitive and selective imaging of latent fingerprints based on enhanced hydrophobic interaction

The demand for in-situ detection of latent fingerprints(LFPs)in ways of high sensitivity,high selectivity,high contrast,low cost and user-friendly is still urgent.To overcome this challenge,a moisture-stable,red-emitting fluoride phosphor K_(3)AlF_(6):Mn^(4+)(KAF:Mn^(4+))with an organic hydrophobic skin was prepared.The phosphor has a uniform and superfine morphology with excellent luminescence properties.More importantly,this non-ultraviolet(UV)or non-near infrared(NIR)induced phosphor was proved to be an ideal fluorescent label for LFP imaging,which is both friendly for touch DNA analysis and compatible to forensic light sources.The well-defined ridge details with little background interference on various surfaces were presented by the oleic acid(OA)modified KAF:Mn^(4+)(KAF:Mn^(4+)-OA)phosphor in few seconds using the powder dusting method.To confirm the high selectivity of KAF:Mn^(4+)-OA for LFP imaging,an efficient quantitative evaluation method is proposed with the aid of ImageJ&Origin software.Due to the superiority of the Mn^(4+)-doped fluoride for the rapid imaging of LFPs in terms of lowcost,high compatibility and good availability,it is expected to be a promising candidate for forensic science as well as fluorescence imaging in other fields instead of rare earth luminescent materials.

Mesencephalic Progenitors Can Differentiate into TH-Positive Neurons and Improve Dysfunction in PD Rats

This study investigated the possibility of using mesencephalic progenitors (MPs) in the treatment of Parkinson's disease (PD). MPs were prepared from E 11 13 rats and proliferated in the serum free medium with basic fibroblast growth factor (bFGF) for 10 days. Cells were then collected and implanted into the striatum only-single grafts, or simultaneously into the substantia nigra (SN) and the striatum-double grafts. Twelve weeks after transplantation, 1,1' dioctadecyl 3,3,3',3' tetramethylindocarbocyanine perchlorate (DiI), a fluorescent dye, was microinjected into the ipsilateral striatum. The results show that double grafted MPs have more potent effects on rotational behavior than single grafted MPs. Injection of the retrograde tracer DiI into the striatum results in fluorescent labeled cells within the intranigral grafts only in double graft subjects. These data suggest that MP transplants not only can improve rotational behavior, but also help to reestablish nigrostriatal connections, so that such transplants may become an efficient way of treating PD.