Artificial Skin with Patterned Stripes for Color Camouflage and Thermoregulation.

Chameleons are famous for their quick color changing abilities, and it is commonly assumed that they do this for camouflage. However, recent reports revealed that chameleons also change color for body temperature regulation. Inspired by the structure of the panther chameleon's skin, a stripe-patterned poly(N-isopropylacrylamide) (PNIPAM) and polyacrylamide (PAM) hydrogel film with a laminated structure is fabricated in this work; thus, both camouflage and thermoregulation can be achieved through controlling Vis and NIR light effectively. For the PNIPAM stripe, the upper layer is the native PNIPAM hydrogel and the lower layer is the carbon nanotube-composited PNIPAM hydrogel. Thus, the PNIPAM stripe is capable of reaching 28 °C at a low environmental temperature (12 °C) and a low radiation intensity (20 mW cm-2), while preventing the body temperature from rising by changing to white under a strong radiation intensity (100 mW cm-2). For the PAM stripe, the upper layer combines colloidal photonic crystals and displays a tunable structural color by stretching, and the lower layer is mixed with PNIPAM microgels for thermal regulation. Through the fabrication of multifunctional patterns, the film can achieve both dynamic structural color and thermoregulation by precisely controlling solar radiation absorption, scattering, and reflection. More importantly, in the stripe-patterned system, the shrinkage of the PNIPAM stripes can effectively trigger the elongation of the PAM stripe, which endows the structural color changing process to be self-powered completely. The performances show that the stripe-patterned film may have potential applications in intelligent coatings, especially in areas with large temperature differences during the day such as high plains.

Bio-Inspired Hydrogel-Elastomer Actuator with Bidirectional Bending and Dynamic Structural Color.

In nature, some creatures can change their body shapes and surface colors simultaneously to respond to the external environments, which greatly inspired researchers in the development of color-tunable soft actuators. In this work, we present a facile method to prepare a smart hydrogel actuator that can bend bidirectionally and change color simultaneously, just like an octopus. The actuator is fabricated by elastomer/hydrogel bilayer and the hydrogel layer was decorated with thermoresponsive microgels as the photonic crystal blocks. Compared with the previously reported poly(N-isopropylacrylamide) hydrogel-based bilayer hydrogel actuators, which are generally limited to one-directional deformation, the elastomer/hydrogel bilayer actuator prepared in our work exhibits unique bidirectional bending behavior in accordance with the change of structural color. The bending degrees can be changed from -360° to 270° in response to solution temperatures ranging from 20 °C to 60 °C. At the same time, the surface color changes from red to green, and then to blue, covering the full visible light spectrum. The bending direction and degree of the hydrogel actuator can easily be adjusted by tuning the layer thickness ratio of the elastomer/hydrogel or the composition of the hydrogel. The color-tunable hydrogel-elastomer actuator reported in this work can achieve both programmable deformations and color-changing highly resembling the natural actuating behaviors of creatures.

Overexpression of OsSnRK1a through a green tissue-specific promoter improves rice yield by accelerating sheath-to-panicle transport of nonstructural carbohydrates and increasing leaf photosynthesis.

The redistribution of nonstructural carbohydrates (NSCs) in rice (Oryza sativa) sheaths contributes greatly to grain filling. Sucrose nonfermenting-1-related protein kinase 1 (SnRK1) regulates sheath-to-panicle transport of NSCs during rice grain filling; however, it is unknown whether elevated activity of SnRK1 in sheaths improves NSC transport and grain filling. Expression of OsSnRK1a is mainly responsible for regulating SnRK1 activity in rice sheaths. Analysis of transgenic rice plants containing the OsSnRK1a promoter::GUS construct indicated that OsSnRK1a is widely expressed in rice. Notably, OsSnRK1a is highly expressed in mesophyll cells of sheaths. Therefore, a green tissue promoter specifically expressed in sheaths and leaf parenchyma cells and phloem tissue was used to over-express OsSnRK1a in japonica rice. The transgenic lines exhibited increased SnRK1a expression and SnRK1 activity in sheaths. The NSC and starch in the transgenic lines and WT all showed accumulation before heading and during the early-filling stage, and declining at the peak filling stage. But the starch and NSC content in transgenic lines was lower than that of WT. Moreover, the transgenic lines showed lower sucrose contents and higher sucrose efflux rates. The accelerated sheath NSC transport improved grain filling, and stimulated panicle development in transgenic lines. SnRK1a expression and SnRK1 activity were also increased in the leaves of transgenic lines, which improved leaf photosynthetic activity and contributed to optimal grain filling and panicle development. These results verify the promotion of high SnRK1 activity in sheath NSC transport, and also provide a new approach to improving sheath NSC transport and rice yield.

Design and preparation of highly active TiO2 photocatalysts by modulating their band structure.

Titanium dioxide photocatalysts with high reduction potential and visible light response hold great promise in photochemical conversion. Here, we used a simple glycine doping method to synthesize novel N-TiO2@C photocatalysts with upward shifted conduction bands and narrowed band gaps as well as inhibited recombination of photoinduced electron-hole pairs. The N-TiO2@C photocatalysts exhibited higher visible light response and remarkably enhanced photocatalytic activity in the production of nicotinamide adenine dinucleotide (NADH) by photomediated reduction of NAD+ without any electron mediator. The yield of NADH was up to 70.3 % far greater than that of the undoped TiO2 (11.3 %), and it stabilized at ca. 60 % after 10 cycles. The viability of coupling NADH regeneration with enzymatic reaction (alcohol dehydrogenase) was established in aldehyde reduction where formaldehyde was specifically reduced to methanol. These findings shed new light on the modulation of the band structure of semiconductors and develop an electron mediator free strategy for NADH-dependent artificial photosynthesis through coupled photocatalytic and enzymatic approaches.

Increasing SnRK1 activity with the AMPK activator A-769662 accelerates seed germination in rice.

Sucrose non-fermenting-1-related protein kinase 1 (SnRK1) plays a key role in rice germination. The small molecule drug, A-769662, activates AMP-activated protein kinase, a mammalian homolog of SnRK1. However, it is unknown whether A-769662 activates SnRK1, thereby affecting germination. SnRK1 in desalted extracts from germinating rice seeds was strongly activated by adding A-769662 in vitro. Applying 50 or 100 µM A-769662 accelerated germination and increased the root length, shoot length, and seedling fresh weight. 50 µM A-769662 treatment increased the catalytic activity and phosphorylation of SnRK1 during germination. Transcriptome analysis and biochemical validation were performed to investigate the mechanism whereby A-769662 treatment promoted rice germination. A-769662 treatment promoted starch hydrolysis by increasing the expression and activity of amylase and inhibited starch biosynthesis by decreasing the expression of OsAGPL2, OsAGPS2a, Wx, and SSIIa. The abscisic acid (ABA) level and gene expression of ABA-induced transcription factors, including OsNF-YC9, OsNF-YC12, OsWRKY24, OsPYL8, OsMKKK62, and OsMKKK63, which reduced the inhibition of germination by ABA were decreased under 50 µM A-769662 treatment. The increased expression of the OsACO3 and OsACO5 genes and increased ethylene levels under A-769662 treatment, which counteracted the inhibition of ABA on germination and, thus, promoted germination. These results demonstrate the activation of A-769662 on SnRK1 and further reveal the regulatory mechanism of A-769662 in rice seed germination and nutrient remobilization.

Sucrose nonfermenting-1-related protein kinase 1 regulates sheath-to-panicle transport of nonstructural carbohydrates during rice grain filling.

The remobilization of nonstructural carbohydrates (NSCs) reserved in rice (Oryza sativa) sheaths is essential for grain filling. This assimilate distribution between plant tissues and organs is determined by sucrose non-fermenting-1-related protein kinase 1 (SnRK1). However, the SnRK1-mediated mechanism regulating the sheath-to-panicle transport of NSCs in rice remains unknown. In this study, leaf cutting treatment was used to accelerate NSC transport in the rice sheaths. Accelerated NSC transport was accompanied by increased levels of OsSnRK1a mRNA expression, SnRK1a protein expression, catalytic subunit phosphorylation of SnRK1, and SnRK1 activity, indicating that SnRK1 activity plays an important role in sheath NSC transport. We also discovered that trehalose-6-phosphate, a signal of sucrose availability, slightly reduced SnRK1 activity in vitro. Since SnRK1 activity is mostly regulated by OsSnRK1a transcription in response to low sucrose content, we constructed an snrk1a mutant to verify the function of SnRK1 in NSC transport. NSCs accumulated in the sheaths of snrk1a mutant plants and resulted in a low seed setting rate and grain weight, verifying that SnRK1 activity is essential for NSC remobilization. Using phosphoproteomics and parallel reaction monitoring, we identified 20 SnRK1-dependent phosphosites that are involved in NSC transport. In addition, the SnRK1-mediated phosphorylation of the phosphosites directly affected starch degradation, sucrose metabolism, phloem transport, sugar transport across the tonoplast, and glycolysis in rice sheaths to promote NSC transport. Therefore, our findings reveal the importance, function, and possible regulatory mechanism of SnRK1 in the sheath-to-panicle transport of NSCs in rice.

Fractional flow reserve for coronary stenosis assessment derived from fusion of intravascular ultrasound and X-ray angiography.

BACKGROUND: Intravascular ultrasound (IVUS) provides good insight into lumen boundary and plaques; however, it is still difficult to detect functionally significant stenosis from IVUS images for the guidance of coronary percutaneous intervention (PCI). This study aimed to develop a novel method to estimate fractional flow reserve (FFR) value for determining the functional significance of coronary artery disease through the fusion of IVUS and X-ray angiographic images. METHODS: We developed a novel approach to 3D vessel reconstruction by integrating IVUS with X-ray angiographic images. Based on the reconstructed geometry and the inlet flow derived from the thrombolysis in myocardial infarction (TIMI) frame count, a simplified fluid dynamics equation was established to compute the pressure drop and IVUS-derived FFR (AccuFFRivus) was subsequently obtained. To validate the feasibility and performance of this IVUS-based FFR method, we performed AccuFFRivus calculations on 32 coronary vessels with invasive FFR as the reference standard. RESULTS: Great correlation (r=0.86, P<0.001) was observed between AccuFFRivus and FFR. The area under the receiver-operating characteristic curve (AUC) was higher for AccuFFRivus than minimal lumen area (MLA, <4 mm2) and diameter stenosis rate (DS% ≥50%) [0.98 (95% CI: 0.86 to 1.0) vs. 0.78 (95% CI: 0.60 to 0.91) and 0.66 (95% CI: 0.47 to 0.82)]. Bland-Altman plot showed a mean difference value of -0.011 (limits of agreement: -0.156 to 0.134). CONCLUSIONS: AccuFFRivus is a novel method for hybridizing IVUS and X-ray angiographic images to identify functionally significant stenosis with FFR ≤0.80. The good diagnostic performance from the initial validation study demonstrates the potential for clinical utilization of physiologically guided decision-making. Further validation is required in future studies with a large number of cases.

Diagnostic Performance of Angiography-Based Fractional Flow Reserve for Functional Evaluation of Coronary Artery Stenosis.

Background: A new method for calculating fraction flow reserve (FFR) without pressure-wire (angiography-derived FFR) based on invasive coronary angiography (ICA) images can be used to evaluate the functional problems of coronary stenosis. Objective: The aim of this study was to assess the diagnostic performance of a novel method of calculating the FFR compared to wire-based FFR using retrospectively collected data from patients with stable angina. Methods: Three hundred patients with stable angina pectoris who underwent ICA and FFR measurement were included in this study. Two ICA images with projections >25° apart at the end-diastolic frame were selected for 3D reconstruction. Then, the contrast frame count was performed in an angiographic run to calculate the flow velocity. Based on the segmented vessel, calculated velocity, and aortic pressure, AccuFFRangio distribution was calculated through the pressure drop equation. Results: Using FFR ≤ 0.8 as a reference, we evaluated AccuFFRangio performance for 300 patients with its accuracy, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). Comparison of AccuFFRangio with wire-measured FFR resulted in an area under the curve (AUC) of 0.954 (per-vessel, p < 0.0001). Accuracy for AccuFFRangio was 93.7% for Pa set from measurement and 87% for Pa = 100 mmHg in this clinical study. Overall sensitivity, specificity, PPV, and NPV for per-vessel were 90, 95, 86.7, 96.3, and 57.5, 97.7, 90.2, 86.3%, respectively. Overall accuracy, sensitivity, specificity, PPV, and NPV for 2-dimensional (2D) quantitative coronary angiography (QCA) were 63.3, 42.5, 70.9, 34.7, and 77.2%, respectively. The average processing time of AccuFFRangio was 4.30 ± 1.87 min. Conclusions: AccuFFRangio computed from coronary ICA images can be an accurate and time-efficient computational tool for detecting lesion-specific ischemia of coronary artery stenosis.

Mechanistic process understanding of the self-assembling behaviour of asymmetric bolaamphiphilic short-peptides and their templating for silica and titania nanomaterials.

Investigation of the self-assembly of peptides is critically important to clarify certain biophysical phenomena, fulfill some biological functions, and construct functional materials. However, it is still a challenge to precisely predict the self-assembled structures of peptides because of their complicated driving forces and various assembling pathways. In this work, to elucidate the effects of noncovalent interactions including hydrogen bonding, molecular geometry, and hydrophobic and electrostatic interactions on the peptide self-assembly, a series of asymmetric bolaamphiphilic short peptides consisting of Ac-EI3K-NH2 (EI3K), Ac-EI4K-NH2 (EI4K), Ac-KI3E-NH2 (KI3E) and Ac-KI4E-NH2 (KI4E) were designed and their self-assembling behaviors at different solution pH values were investigated systematically. The peptides self-assembled into twisted nanofibers under most conditions except for EI4K in a strongly alkaline solution and KI4E under a strongly acidic condition, in which they self-assembled into nanotubes via helical monolayer nanosheet intermediates. In particular, KI4E nanotubes are formed under acidic conditions, and its diameters are ∼500 nm much greater than most of the self-assembled structures from bolaamphiphilic peptides. Moreover, reversible morphological transition between the nanotubes and twisted nanofibers was observed with the change in solution pH. Such tunable self-assembled structures and switchable surface properties of the asymmetric bolaamphiphilic short-peptides allow them to be used as templates to construct advanced materials. Silica and titania nanomaterials faithful to the peptide templates in morphology were prepared at ambient temperature. This work clearly elucidates the effects of noncovalent interactions on the peptide self-assembly and also provides new insights into the design and preparation of complicated inorganic materials from tunable organic templates.

Diagnostic accuracy of coronary computed tomography angiography-derived fractional flow reserve.

BACKGROUND: Fractional flow reserve (FFR) is a widely used gold standard to evaluate ischemia-causing lesions. A new method of non-invasive approach, termed as AccuFFRct, for calculating FFR based on coronary computed tomography angiography (CCTA) and computational fluid dynamics (CFD) has been proposed. However, its diagnostic accuracy has not been validated. OBJECTIVES: This study sought to present a novel approach for non-invasive computation of FFR and evaluate its diagnostic performance in patients with coronary stenosis. METHODS: A total of 54 consecutive patients with 78 vessels from a single center who underwent CCTA and invasive FFR measurement were retrospectively analyzed. The CT-derived FFR values were computed using a novel CFD-based model (AccuFFRct, ArteryFlow Technology Co., Ltd., Hangzhou, China). Diagnostic performance of AccuFFRct and CCTA in detecting hemodynamically significant coronary artery disease (CAD) was evaluated using the invasive FFR as a reference standard. RESULTS: Diagnostic accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for AccuFFRct in detecting FFR ≤ 0.8 on per-patient basis were 90.7, 89.5, 91.4, 85.0 and 94.1%, respectively, while those of CCTA were 38.9, 100.0, 5.71, 36.5 and 100.0%, respectively. The correlation between AccuFFRct and FFR was good (r = 0.76 and r = 0.65 on per-patient and per-vessel basis, respectively, both p < 0.0001). Area under the curve (AUC) values of AccuFFRct for identifying ischemia per-patient and per-vessel basis were 0.945 and 0.925, respectively. There was much higher accuracy, specificity and AUC for AccuFFRct compared with CCTA. CONCLUSIONS: AccuFFRct computed from CCTA images alone demonstrated high diagnostic performance for detecting lesion-specific ischemia, it showed superior diagnostic power than CCTA and eliminated the risk of invasive tests, which could be an accurate and time-efficient computational tool for diagnosing ischemia and assisting clinical decision-making.

Programmable Color in a Free-Standing Photonic Microgel Film with Ultra-Fast Response.

In this work, a free-standing microgel film with programmable and angle-independent structural color is prepared via a simple but effective method. Dried poly(styrene-N-isopropylacrylamide-acrylic acid) (pStNIPAAmAA) microgels were stabilized by inter-microgel crosslinking, and thus, only microgels were used to build the optical hydrogel. The free-standing microgel film displayed tunable structural color by the swelling/deswelling of the microgels under external stimuli, such as temperature, pH, ionic strength, and organic solvent. Moreover, the structural color of the film is angle-independent for the disordered microgel arrays. It is worth noting that programmable color stripes which have the panther chameleon's ability to change skin color are successfully fabricated by patterning microgels with different thermoresponsivities. More importantly, the microgel film has an ultrafast response to temperature (1.41 s from 20 to 40 °C) and pH (2.24 s from pH 8.3 to pH 2.0), much faster than that of most optical materials reported in previous studies.

Bioinspired construction of light-harvesting antenna via hierarchically co-assembling approach.

Biomimetic construction of artificial photosystem capable of converting light energy to chemical energy is a promising strategy in solving the increasing serious energy and environmental problems. Herein, we present a new strategy to construct light-harvesting antenna via hierarchical co-assembly of short-peptide and porphyrin and subsequent self-metallization process. The hierarchically organized antenna exhibits both excellent photocatalytic performance and remarkable sustainability under strong light irradiation (35000 lx) and extraordinary sensitivity to weak light (700 lx). In such cases, light energy can be converted into chemical energy and stored in the energy-storage molecules (nicotinamide adenine dinucleotide, NADH) even under weak light irradiation. This provides a promising step towards an artificial photosystem that can utilize weak light. Moreover, the structures and properties of the antenna are dependent on the competition of short-peptide self-assembling and co-assembling with porphyrin molecules and can be regulated by their molar ratio. This provides new insights into the design and construction of light-harvesting antennas with integrated functionality via precise control of pigments aggregation and coupling of different functional units.

Peptide-Mediated Synthesis of Zeolitic Imidazolate Framework-8 with Controllable Morphology and Size.

Peptides with a sequence of Nap-Ix-GPLGLAG-R4-NH2 (x = 2, 4, and 6, shorted as I2R4, I4R4, and I6R4) were used as capping agents for the synthesis of zeolitic imidazolate framework-8 (ZIF-8) in water. Peptide addition can significantly inhibit the growth of ZIF-8 crystals. The shape and size of ZIF-8 crystals was related closely to the number of isoleucine (Ile, I) residues as well as concentration of the peptide. The shape of ZIF-8 crystals changes from rhomboid dodecahedron to truncated rhombic dodecahedron to cube with the decreasing number of isoleucine residues from six to two. At a peptide concentration of 1.0 mM, the morphology of ZIF-8 crystals was cubic, truncated rhombic dodecahedron, and typical rhombic dodecahedron in the cases of I2R4, I4R4, and I6R4, respectively. Also, the particle size can be regulated from ca. 1.7 µm to <100 nm by controlling the peptide concentration from 0 to 2.0 mM. This work develops a simple and green method for the synthesis of ZIF-8 crystals with controllable shape and size in water, which shows high potential for biomedical and biological applications.

Peptide-mediated porphyrin based hierarchical complexes for light-to-chemical conversion.

We present a new strategy for the biomimetic preparation of integrated photoactive complexes consisting of light harvesting and electron separation/transfer components via the hierarchical assembly of porphyrin and platinum nanoparticles on the surface of short-peptide self-assembled structures. The complexes can catalyze the conversion of visible light energy into chemical energy in the absence of an electron mediator and store it as nicotinamide adenine dinucleotide (NADH). This provides a promising step towards artificial photosystems through precisely controlled interactions of light-capturing agents, electron separators and biomolecules.

Poly(N-isopropylacrylamide)-Based Thermoresponsive Composite Hydrogels for Biomedical Applications.

Poly(N-isopropylacrylamide) (PNIPAM)-based thermosensitive hydrogels demonstrate great potential in biomedical applications. However, they have inherent drawbacks such as low mechanical strength, limited drug loading capacity and low biodegradability. Formulating PNIPAM with other functional components to form composited hydrogels is an effective strategy to make up for these deficiencies, which can greatly benefit their practical applications. This review seeks to provide a comprehensive observation about the PNIPAM-based composite hydrogels for biomedical applications so as to guide related research. It covers the general principles from the materials choice to the hybridization strategies as well as the performance improvement by focusing on several application areas including drug delivery, tissue engineering and wound dressing. The most effective strategies include incorporation of functional inorganic nanoparticles or self-assembled structures to give composite hydrogels and linking PNIPAM with other polymer blocks of unique properties to produce copolymeric hydrogels, which can improve the properties of the hydrogels by enhancing the mechanical strength, giving higher biocompatibility and biodegradability, introducing multi-stimuli responsibility, enabling higher drug loading capacity as well as controlled release. These aspects will be of great help for promoting the development of PNIPAM-based composite materials for biomedical applications.

Cell Adhesion and Migration Behaviors on Patterned Thermoresponsive Microgel Stripes.

Cell adhesion and migration behaviors are influenced by their surrounding environments. In this work, patterned poly(N-isopropylacrylamide-styrene) (pNIPAAmSt) microgel stripes, which can provide a more complex environment, were fabricated on polyethyleneimine (PEI)-precoated glass substrate, and their effects on cell adhesion and migration behavior were investigated. The results showed that patterned cell layers can be formed on the space regions either by selective adhesion of the cells or by detaching the attached cells from the thermoresponsive microgel stripes via mild temperature stimuli. The migration behavior of the cells was affected by both the cell types and the width of the microgel stripes. The wider the microgel stripe, the harder it is for the cells to migrate, and the longer the patterned cell layers can be conserved. It is worth noting that the cell-cell interaction in the cocultured system plays a key role in the cell migration process. This work may offer a platform to study the adhesion and migration behavior of mono/cocultured cells in a more controllable system.

Fabrication of Patterned Thermoresponsive Microgel Strips on Cell-Adherent Background and Their Application for Cell Sheet Recovery.

Interfaces between materials and cells play a critical role in cell biomedical applications. Here, a simple, robust, and cost-effective method is developed to fabricate patterned thermoresponsive poly(N-isopropylacrylamide-co-styrene) microgel strips on a polyethyleneimine-precoated, non-thermoresponsive cell-adherent glass coverslip. The aim is to investigate whether cell sheets could be harvested from these cell-adherent surfaces patterned with thermoresponsive strips comprised of the microgels. We hypothesize that if the cell-to-cell interaction is strong enough to retain the whole cell sheet from disintegration, the cell segments growing on the thermoresponsive strips may drag the cell segments growing on the cell-adherent gaps to detach, ending with a whole freestanding and transferable cell sheet. Critical value concerning the width of the thermoresponsive strip and its ratio to the non-thermoresponsive gap may exist for cell sheet recovery from this type of surface pattern. To obtain this critical value, a series of strip patterns with various widths of thermoresponsive strip and non-thermoresponsive gap were prepared using negative microcontact printing technology, with COS7 fibroblast cells being used to test the growth and detachment. The results unraveled that COS7 cells preferentially attached and proliferated on the cell-adherent, non-thermoresponsive gaps to form patterned cell layers and that they subsequently proliferated to cover the microgel strips to form a confluent cell layer. Intact COS7 cell sheets could be recovered when the width of the thermoresponsive strip is no smaller than that of the non-thermoresponsive gap. Other cells such as HeLa, NIH3T3, 293E, and L929 could grow similarly; that is, they showed initial preference to the non-thermoresponsive gaps and then migrated to cover the entire patterned surface. However, it was difficult to detach them as cell sheets due to the weak interactions within the cell layers formed. In contrast, when COS7 and HeLa cells were cultured successively, they formed the cocultured cell layer that could be detached together. These freestanding patterned cell sheets could lead to the development of more elaborate tumor models for drug targeting and interrogation.

Visible and Near-Infrared Dual-Emission Carbogenic Small Molecular Complex with High RNA Selectivity and Renal Clearance for Nucleolus and Tumor Imaging.

Fluorescence imaging requires bioselective, sensitive, nontoxic molecular probes to detect the precise location of lesions for fundamental research and clinical applications. Typical inorganic semiconductor nanomaterials with large sizes (>10 nm) can offer high-quality fluorescence imaging due to their fascinating optical properties but are limited to low selectivity as well as slow clearance pathway. We here report an N- and O-rich carbogenic small molecular complex (SMC, MW < 1000 Da) that exhibits high quantum yield (up to 80%), nucleic acid-binding enhanced excitation-dependent fluorescence (EDF), and a near-infrared (NIR) emission peaked at 850 nm with an ultralarge Stokes shift (∼500 nm). SMCs show strong rRNA affinity, and the resulting EDF enhancement allows multicolor visualization of nucleoli in cells for clear statistics. Furthermore, SMCs can be efficiently accumulated in tumor in vivo after injection into tumor-bearing mice. The NIR emission affords high signal/noise ratio imaging for delineating the true extent of tumor. Importantly, about 80% of injected SMCs can be rapidly excreted from the body in 24 h. No appreciable toxicological responses were observed up to 30 days by hematological, biochemical, and pathological examinations. SMCs have great potential as a promising nucleolus- and tumor-specific agent for medical diagnoses and biomedical research.

An Ultra-High Fluorescence Enhancement and High Throughput Assay for Revealing Expression and Internalization of Chemokine Receptor CXCR4.

Revealing chemokine receptor CXCR4 expression, distribution, and internalization levels in different cancers helps to evaluate cancer progression or prognosis and to set personalized treatment strategy. We here describe a sensitive and high-throughput immunoassay for determining CXCR4 expression and distribution in cancer cells. The assay is accessible to a wide range of users in an ordinary lab only by dip-coating poly(styrene-co-N-isopropylacrylamide) spheres on the glass substrate. The self- assembled spheres form three-dimensional photonic colloidal crystals which enhance the fluorescence of CF647 and Alexa Fluor 647 by a factor of up to 1000. CXCR4 in cells is detected by using the sandwich immunoassay, where the primary antibody recognizes CXCR4 and the secondary antibody is labeled with CF647. With the newly established assay, we quantified the total expression of CXCR4, its distribution on the cell membrane and cytoplasm, and revealed their internalization level upon SDF-1α activation in various cancer cells, even for those with extremely low expression level.

Patterned Thermoresponsive Microgel Surfaces to Control Cell Detachment.

The aim of this work is to examine how adhered individual cells could detach from the patterned, discontinuous thermoresponsive coating substrate and how different patterns in the form of thermoresponsive squares and gaps would affect cell detachment. Microgels prepared from copolymerization of N-isopropylacrylamide and styrene (pNIPAAmSt) were spin-coated on polyethylenimine (PEI) precoated glass coverslips to form a uniform microgel monolayer; then a surface-moisturized PMDS stamp was used to contact the microgel monolayer at room temperature. The thin layer of water on the PDMS stamp surface worked as an ink to penetrate the microgels so that any microgels in direct contact with the wet stamp surface became swollen and could be peeled away, while uncontacted microgels formed patterns. Using this method, various patterns with different thermo-island diameters and gaps could be fabricated. NIH3T3 fibroblast cells were then cultured on these patterns to study their detachment behavior. It was found that cells could detach not only from these discontinuous thermoresponsive coatings, but also from the patterned surfaces with the thermoresponsive area being as low as 20% of the cell spread area.