Design of Optical System for Ultra-Large Range Line-Sweep Spectral Confocal Displacement Sensor.

The spectrum confocal displacement sensor is an innovative type of photoelectric sensor. The non-contact advantages of this method include the capacity to obtain highly accurate measurements without inflicting any harm as well as the ability to determine the object's surface contour recovery by reconstructing the measurement data. Consequently, it has been widely used in the field of three-dimensional topographic measuring. The spectral confocal displacement sensor consists of a light source, a dispersive objective, and an imaging spectrometer. The scanning mode can be categorized into point scanning and line scanning. Point scanning is inherently present when the scanning efficiency is low, resulting in a slower measurement speed. Further improvements are necessary in the research on the line-scanning type. It is crucial to expand the measurement range of existing studies to overcome the limitations encountered during the detection process. The objective of this study is to overcome the constraints of the existing line-swept spectral confocal displacement sensor's limited measuring range and lack of theoretical foundation for the entire system. This is accomplished by suggesting an appropriate approach for creating the optical design of the dispersive objective lens in the line-swept spectral confocal displacement sensor. Additionally, prism-grating beam splitting is employed to simulate and analyze the imaging spectrometer's back end. The combination of a prism and a grating eliminates the spectral line bending that occurs in the imaging spectrometer. The results indicate that a complete optical pathway for the line-scanning spectral confocal displacement sensor has been built, achieving an axial resolution of 0.8 µm, a scanning line length of 24 mm, and a dispersion range of 3.9 mm. This sensor significantly expands the range of measurements and fills a previously unaddressed gap in the field of analyzing the current stage of line-scanning spectral confocal displacement sensors. This is a groundbreaking achievement for both the sensor itself and the field it operates in. The line-scanning spectral confocal displacement sensor's design addresses a previously unmet need in systematic analysis by successfully obtaining a wide measuring range. This provides systematic theoretical backing for the advancement of the sensor, which has potential applications in the industrial detection of various ranges and complicated objects.

N4 -Allyldeoxycytidine: A New DNA Tag with Chemical Sequencing Power for Pinpointing Labelling Sites, Mapping Epigenetic Markers, and In Situ Imaging.

DNA tagging with base analogues has found numerous applications. To precisely record the DNA labelling information, it would be highly beneficial to develop chemical sequencing tags that can be encoded into DNA as regular bases and decoded as mutant bases following a mild, efficient and bioorthogonal chemical treatment. Here we reported such a DNA tag, N4 -allyldeoxycytidine (a4 dC), for labeling and identifying DNA by in vitro assays. The iodination of a4 dC led to fast and complete formation of 3, N4 -cyclized deoxycytidine, which induced base misincorporation during DNA replication and thus could be located at single base resolution. We explored the applications of a4 dC in pinpointing DNA labelling sites at single base resolution, mapping epigenetic marker N4 -methyldeoxycytidine, and imaging nucleic acids in situ. In addition, mammalian cellular DNA could be metabolically labelled with a4 dC. Our study sheds light on the design of next generation DNA tags with chemical sequencing power.

A metabolic labeling method detects m6A transcriptome-wide at single base resolution.

Transcriptome-wide mapping of N6-methyladenosine (m6A) at base resolution remains an issue, impeding our understanding of m6A roles at the nucleotide level. Here, we report a metabolic labeling method to detect mRNA m6A transcriptome-wide at base resolution, called 'm6A-label-seq'. Human and mouse cells could be fed with a methionine analog, Se-allyl-L-selenohomocysteine, which substitutes the methyl group on the enzyme cofactor SAM with the allyl. Cellular RNAs could therefore be metabolically modified with N6-allyladenosine (a6A) at supposed m6A-generating adenosine sites. We pinpointed the mRNA a6A locations based on iodination-induced misincorporation at the opposite site in complementary DNA during reverse transcription. We identified a few thousand mRNA m6A sites in human HeLa, HEK293T and mouse H2.35 cells, carried out a parallel comparison of m6A-label-seq with available m6A sequencing methods, and validated selected sites by an orthogonal method. This method offers advantages in detecting clustered m6A sites and holds promise to locate nuclear nascent RNA m6A modifications.

A Mutation-Based Method for Pinpointing a DNA N6 -Methyladenine Methyltransferase Modification Site at Single Base Resolution.

DNA N6 -methyladenine (6mA) has recently received notable attention due to an increased finding of its functional roles in higher eukaryotes. Here we report an enzyme-assisted chemical labeling method to pinpoint the DNA 6mA methyltransferase (MTase) substrate modification site at single base resolution. A designed allyl-substituted MTase cofactor was applied in the catalytic transfer reaction, and the allyl group was installed to the N6 -position of adenine within a specific DNA sequence to form N6 -allyladenine (6aA). The iodination of 6aA allyl group induced the formation of 1, N6 -cyclized adenine which caused mutations during DNA replication by a polymerase. Thus the modification site could be precisely detected by a mutation signal. We synthesized 6aA deoxynucleoside and deoxynucleotide model compounds and a 6aA-containing DNA probe, and screened nine DNA polymerases to define an optimal system capable of detecting the substrate modification site of a DNA 6mA MTase at single-base resolution.

Precise identification of an RNA methyltransferase's substrate modification site.

Here we report a simple and nonradioactive biochemical assay which is capable of accurately determining the substrate methylation sites of human RNA N6-methyladenosine methyltransferases METTL3/METTL14 and METTL16. This method employs enzyme-assisted chemical labelling of a specific base in an RNA substrate with the assistance of an allyl-substituted methyltransferase cofactor, and enables precise identification of the labelling site by a mutation signal from standard nucleic acid sequencing. Our method provides a platform to investigate the enzymatic methylations of long and structurally complex RNA substrates, and facilitates the discovery of new methyltransferases.

Design, synthesis, and biological evaluation of novel tetrahydroisoquinoline derivatives as potential antitumor candidate.

A novel class of tetrahydroisoquinoline derivatives was designed and synthesized as antitumor agents and evaluated for their in vitro and in vivo biological activities. The antiproliferative activities of all the target compounds on HUVEC, MCF-7, and HT-29 were evaluated. Compared with colchicine (1.04 × 10-2  µm), 17d and 17e exhibited outstanding activity on MCF-7 with IC50 values 0.26 × 10-2  µm and 0.89 × 10-3 µm in cell cytotoxicity assay. The tubulin polymerization assay demonstrated that 17d and 17e exhibited better inhibition rate. In the MCF-7-xenograft mouse model that was treated with 17d and 17e by intraperitoneal injection, the tumor weight was decreased at same rate with tamoxifen, and relative tumor proliferation rates were 59.48% and 41.33%, while tamoxifen was 45.08% with a daily dose of 20 mg/kg, which were demonstrated potent in vivo efficacy.