Rhodopsin, composed of opsin and isomeric retinal, acts as the primary photoreceptor by converting light into electric signals. Inspired by rhodopsin, we have fabricated a light-regulated ionic gate on the basis of the design of a graphene oxide (GO)-biomimetic DNA-nanochannel architecture. In this design, photoswitchable azobenzene (Azo)-DNA is introduced to the surface of porous anodic alumina (PAA) membrane. With modulation of the interaction between the GO blocker and Azo-DNA via flexibly regulating trans and cis states of Azo under the irradiation of visible and ultraviolet light, alternatively, the ionic gate is switched between ON and OFF states. This newly constructed ionic gate can possess high efficiency for the control of ion transport because of the high blocking property of GO and the rather tiny path within the barrier layer which are both first employed to fabricate ionic gate. We anticipate that this rhodopsin-like ionic gate may provide a new model and method for the investigation of ion channel, ion function, and ion quantity. In addition, because of the advantages of simple fabrication, good biocompatibility, and universality, this bioinspired system may have potential applications as optical sensors, in photoelectric transformation, and in controllable drug delivery.

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With hydrophilic surface and high density of functional groups, MXene can efficiently adsorb single-stranded DNA to enhance target-induced strand release and quench the fluorescence. Herein, MXene is coupled with CRISPR-Cas12a to sensitively detect LPS and bacteria. Specifically, the aptamer is well designed to initiate the trans-cleavage activity of CRISPR-Cas12a to indiscriminately cleave single-stranded DNA, resulting it to be far away from MXene and the recovery of fluorescence. The target can effectually induce the release of the aptamer strand from the hybrid duplex with the assistance of MXene. The formed aptamer/target complex will inhibit the activation of CRISPR-Cas12a and its trans-cleavage on single-stranded DNA. The established method can selectively and sensitively quantify LPS and Gram-negative bacteria in different samples with detection limits of 11 pg/mL and 23 CFU/mL, respectively. Our study provides a new insight for exploration of universal analytical methods based on MXene coupled with CRISPR-Cas12a.

Rapid and accurate identification of semen is critical for male infertility diagnosis and the arrangement of personalized treatment. However, the complexity and diversity of samples impose lots of restrictions in detection. To solve this problem, we propose a colorimetric sensor array in this work by coupling zirconium metal–organic frameworks (Zr-MOFs) with single-stranded-DNA-decorated gold nanoparticles (ssDNA-AuNPs) for human semen identification. Because of the coordination interactions between the Zr6 clusters and the DNA phosphate backbone, as well as π–π stacking and H-bonding, Zr-MOFs can absorb and precipitate AuNPs with the aid of single-stranded DNA. What’s more, addition of semen samples in the test solution, proteins, or other contents in the samples will affect the co-precipitation of Zr-MOFs and ssDNA-AuNPs. Subsequently, the color of the supernatant will change and a method to identify human semen can be developed. Further studies reveal that the method can completely detect different semen cases based on the differences in inclusions, demonstrating the characteristics of simplicity, feasibility, and sensitivity in the application of male infertility diagnosis.

ABSTRACT. The cytoskeleton mediates various cellular processes such as differentiation and fusion, including in the filopodia and podosomes. However, apart from cell migration and formation of the sealing zone, little is known regarding the changes and related regulatory mechanisms of the cytoskeleton and additional roles of the filopodia and podosomes during the differentiation and fusion of osteoclasts. The cytomorphology and cytoskeleton of osteoclasts in the differentiation process were evaluated using tartrate-resistant acid phosphatase staining and immunofluorescence staining. Moreover, the expression levels of Rho GTPases and enzymes related to osteoclast differentiation and bone resorption were detected by quantitative reverse transcription-polymerase chain reaction. We detected 3 types of filopodia in osteoclast precursors and only 1 type of filopodia in undifferentiated cells. Mature osteoclasts were completely devoid of filopodia. Interestingly, cell fusion was highly specific, and the fusion New roles of filopodia and podosomes of osteoclasts initially occurred to the filopodia. Confocal images revealed that F-actin and microtubules significantly differed among fused cells. These results suggest that filopodia and podosomes not only play important roles in cell migration and the formation of sealing zones but also in the prefusion selectivity of 2 cells and the movement direction of the cell nucleus and cytoplasm during the fusion process. In addition, cdc42v1, RhoU, and RhoF regulate the formation of 3 types of filopodia during various stages of differentiation, while Rac1, Rac2, and filament A may be associated with cell selectivity during the fusion process.

Fluorescence imaging tools enable the in situ visualization of those molecules involved in various cell signaling pathways, but directly describing these pathways instead of the separate mediators in situ is much more meaningful and still full of challenges. In this work, a dualresponsive DNA nanodevice that allows the available imaging of an apoptotic signaling pathway in living cells has been developed. The nanodevice is constructed through assembling an elaborately designed Y-shaped DNA (Y-DNA) layer on gold nanoparticles (AuNPs). Only if an apoptotic signaling pathway involving the manganese superoxide dismutase (MnSOD) mRNA and downstream cytochrome c (Cyt c) is presented to serve as the input, the nanodevice can perform an "AND" logic gate operation, disassembling the Y-DNA from the AuNP surface and thereafter outputting a fluorescence signal. In comparison with the fluorescence imaging methods that target individual specific molecules, our strategy allows direct profiling of a specific signaling pathway by connected characterization of two correlative targets. The programmable feature of this strategy also shows the potential for the profiling of other signaling pathways. The concept of studying the line connecting two points will contribute to the systematic interrogation on the signaling networks in situ as the networks are composed of lines instead of individual points.

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Sustained expression of the GH gene has been shown to have detrimental effects on the health of animals. In the current study, transgenic founder pigs, with controllable pig growth hormone (pGH) expression, were cloned via the handmade cloning method (HMC), and pGH expression levels were examined at the cellular and organismal levels. The serum pGH levels in 3 founder male pigs were found to be significantly higher after induction with intramuscular injection of doxycycline (DOX) compared to baseline. A daily dose of DOX was administered via feed to these animals for a period of 65 to 155 days. The growth rate, feed efficiency and pGH serum concentration increased in the DOX-induced transgenic group compared with the other groups. 8 numbers of animals were euthanized and the dressing percentage, loin muscle and lean meat percentage were significantly higher in the DOX-induced F1 transgenic group compared with the other groups. In this study a large population of transgenic pigs, with integrated controllable expression of a transgene, was obtained. The transgenic pigs were healthy and normal in terms of reproductive capability. At the same time, feed efficiency was improved, production processes were accelerated and meat yield was increased.