28 research outputs found

    GE11-antigen-loaded hepatitis B virus core antigen virus-like particles efficiently bind to TNBC tumor

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    PurposeThis study aimed to explore the possibility of utilizing hepatitis B core protein (HBc) virus-like particles (VLPs) encapsulate doxorubicin (Dox) to reduce the adverse effect caused by its off-target and toxic side effect.MethodsHere, a triple-negative breast cancer (TNBC) tumor-targeting GE11-HBc VLP was constructed through genetic engineering. The GE11 peptide, a 12-amino-acid peptide targeting epidermal growth factor receptor (EGFR), was inserted into the surface protein loops of VLPs. The Dox was loaded into HBc VLPs by a thermal-triggered encapsulation strategy. The in vitro release, cytotoxicity, and cellular uptake of TNBC tumor-targeting GE11-HBc VLPs was then evaluated.ResultsThese VLPs possessed excellent stability, DOX loading efficiency, and preferentially released drug payload at high GSH levels. The insertion of GE11 targeting peptide caused improved cellular uptake and enhanced cell viability inhibitory in EGFR high-expressed TNBC cells.ConclusionTogether, these results highlight DOX-loaded, EGFR-targeted VLPs as a potentially useful therapeutic choice for EGFR-overexpressing TNBC

    Low-Cost Chlorophyll Fluorescence Imaging System Applied in Plant Physiology Status Detection

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    ObjectiveChlorophyll fluorescence (ChlF) emission from photosystem II (PSII) is closely coupled with photochemical reactions. As an efficient and non-destructive means of obtaining plant photosynthesis efficiency and physiological state information, the collection of fluorescence signals is often used in many fields such as plant physiological research, smart agricultural information sensing, etc. Chlorophyll fluorescence imaging systems, which is the experimental device for collecting the fluorescence signal, have difficulties in application due to their high price and complex structure. In order to solve the issues, this paper investigates and constructs a low-cost chlorophyll fluorescence imaging system based on a micro complementary metal oxide semiconductor (CMOS) camera and a smartphone, and carries out experimental verifications and applications on it.MethodThe chlorophyll fluorescence imaging system is mainly composed of three parts: excitation light, CMOS camera and its control circuit, and a upper computer based on a smartphone. The light source of the excitation light group is based on the principle and characteristics of chlorophyll fluorescence, and uses a blue light source of 460 nm band to achieve the best fluorescence excitation effect. In terms of structure, the principle of integrating sphere was borrowed, the bowl-shaped light source structure was adopted, and the design of the LED surface light source was used to meet the requirements of chlorophyll fluorescence signal measurement for the uniformity of the excitation light field. For the adjustment of light source intensity, the control scheme of pulse width modulation was adopted, which could realize sequential control of different intensities of excitation light. Through the simulation analysis of the light field, the light intensity and distribution characteristics of the light field were stuidied, and the calibration of the excitation light group was completed according to the simulation results. The OV5640 micro CMOS camera was used to collect fluorescence images. Combined with the imaging principle of the CMOS camera, the fluorescence imaging intensity of the CMOS camera was calculated, and its ability to collect chlorophyll fluorescence was analyzed and discussed. The control circuit of the CMOS camera uses an STM32 microcontroller as the microcontroller unit, and completes the data communication between the synchronous light group control circuit and the smartphone through the RS232 to TTL serial communication module and the full-speed universal serial bus, respectively. The smartphone upper computer software is the operating software of the chlorophyll fluorescence imaging system user terminal and the overall control program for fluorescence image acquisition. The overall workflow could be summarized as the user sets the relevant excitation light parameters and camera shooting instructions in the upper computer as needed, sends the instructions to the control circuit through the universal serial bus and serial port, and completes the control of excitation light and CMOS camera image acquisition. After the chlorophyll fluorescence image collection was completed, the data would be sent back to the smart phone or server for analysis, processing, storage, and display. In order to verify the design of the proposed scheme, a prototype of the chlorophyll fluorescence imaging system based on this scheme was made for experimental verification. Firstly, the uniformity of the light field was measured on the excitation light to test the actual performance of the excitation light designed in this article. On this basis, a chlorophyll fluorescence imaging experiment under continuous light excitation and modulated pulse light protocols was completed. Through the analysis and processing of the experimental results and comparison with mainstream chlorophyll fluorometers, the fluorescence imaging capabilities and low-cost advantages of this chlorophyll fluorometer were further verified.Results and DiscussionsThe maximum excitation light intensity of the chlorophyll fluorescence imaging system designed in this article was 6250 µmol/(m2·s). Through the simulation analysis of the light field and the calculation and analysis of the fluorescence imaging intensity of the CMOS camera, the feasibility of collecting chlorophyll fluorescence images by the OV5640 micro CMOS camera was demonstrated, which provided a basis for the specific design and implementation of the fluorometer. In terms of hardware circuits, it made full use of the software and hardware advantages of smartphones, and only consisted of the control circuits of the excitation light and CMOS camera and the corresponding communication modules to complete the fluorescence image collection work, simplifying the circuit structure and reducing hardware costs to the greatest extent. The final fluorescence instrument achieved a collection resolution of 5 million pixels, a spectral range of 400~1000 nm, and a stable acquisition frequency of up to 42 f/s. Experimental results showed that the measured data was consistent with theoretical analysis and simulation, which could meet the requirements of fluorescence detection. The instrument was capable of collecting images of chlorophyll fluorescence under continuous light excitation or the protocol of modulated pulsed light. The acquired chlorophyll fluorescence images could reflect the two-dimensional heterogeneity of leaves and could effectively distinguish the photosynthetic characteristics of different leaves. Typical chlorophyll fluorescence parameter images of Fv/Fm, Rfd, etc. were in line with expectations. Compared with the existing chlorophyll fluorescence imaging system, the chlorophyll fluorescence imaging system designed in this article has obvious cost advantages while realizing the rapid detection function of chlorophyll fluorescence.ConclusionsThe instrument is with a simple structure and low cost, and has good application value for the detection of plant physiology and environmental changes. The system is useful for developing other fluorescence instruments

    Methylation variation of N<sub>5</sub>CG in human and mouse cells.

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    <p><b>(A)</b> Human brain samples. <b>(B)</b> Mouse brain cells. <b>(C)</b> Human ESCs and iPSCs. <b>(D)</b> Human normal somatic cells. <b>(E)</b> Human PGCs. <b>(F)</b> Human gonadal somatic cells (SOMAs).</p

    Methylation variation (above) and demethylation variation (below) in human brain cells of difference sequencing depth.

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    <p>Methylation variation (above) and demethylation variation (below) in human brain cells of difference sequencing depth.</p

    The bimodal distribution of CpG methylation in human brain samples.

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    <p><b>(A)</b> The bimodal bias caused by the inhomogeneity of sequencing depth in human brain cells (chromosome 1 of the 12yr sample). <b>(B)</b> The observed ratio and the random ratio of 0 and 1 end of the methylation level distribution in different sequencing depths. <b>(C)</b> The observed and random methylation level distribution of all sequencing depth (black circle and red circle respectively) and the observed and random methylation level distribution under the sequencing depth of 16x is in green triangle and blue triangle respectively.</p

    Multifunctional Freestanding Microprobes for Potential Biological Applications

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    Deep-level sensors for detecting the local temperatures of inner organs and tissues of an animal are rarely reported. In this paper, we present a method to fabricate multifunctional micro-probes with standard cleanroom procedures, using a piece of stainless-steel foil as the substrate. On each of the as-fabricated micro-probes, arrays of thermocouples made of Pd–Cr thin-film stripes with reliable thermal sensing functions were built, together with Pd electrode openings for detecting electrical signals. The as-fabricated sword-shaped freestanding microprobes with length up to 30 mm showed excellent mechanical strength and elastic properties when they were inserted into the brain and muscle tissues of live rats, as well as suitable electrochemical properties and, therefore, are promising for potential biological applications

    Methylation variation of N<sub>5</sub>CGA (upper left), N<sub>5</sub>CGC (upper right), N<sub>5</sub>CGG (lower left) and N<sub>5</sub>CGT (lower right) of chromosome 1 in human brain cells.

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    <p>The <i>KL</i>(0) of ACGN<sub>3</sub>, CCGN<sub>3</sub>, GCGN<sub>3</sub> and TCGN<sub>3</sub> are represented as black circle, red square, green diamond and blue triangle, respectively. N<sub>5</sub>, N<sub>3</sub> = A, C, G or T.</p
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