Molecular cloning and functional characterization of BcTSA in the biosynthesis of indole alkaloids in Baphicacanthus cusia

Baphicacanthus cusia (Nees) Bremek (B. cusia) is an essential traditional Chinese herb that is commonly used to treat colds, fever, and influenza. Indole alkaloids, such as indigo and indirubin, are the primary active constituents of B. cusia. The indole-producing reaction is crucial for regulating the flow of indole alkaloids metabolites along the pathways and coordinating primary and secondary product biosynthesis in plants. The tryptophan synthase alpha-subunit (TSA) can catalyse a process that produces indole, which is free to enter secondary metabolite pathways; however, the underlying potential mechanism of regulating indigo alkaloids synthesis remains unknown. Here, a BcTSA was cloned from the transcriptome of B. cusia. The BcTSA has a significant degree of similarity with other plant TSAs according to bioinformatics and phylogenetic analyses. Quantitative real-time PCR (RT-qPCR) research showed that BcTSA was dramatically enhanced in response to treatment with methyl jasmonate (MeJA), salicylic acid (SA), and abscisic acid (ABA), and was predominantly expressed in the stems as opposed to the leaves and rhizomes. Subcellular localization revealed that BcTSA is localized in chloroplasts, which is compatible with the fact that the conversion of indole-3-glycerol phosphate (IGP) to indole occurs in chloroplasts. The complementation assay results showed that BcTSA was functional, demonstrating that it was capable of catalyzing the conversion of IGP to indole. BcTSA was shown to stimulate the manufacture of indigo alkaloids including isatin, indigo, and indirubin when the gene was overexpressed in the hairy roots of Isatis indigotica. In conclusion, our research provides novel perspectives that might be applied to manipulating the indole alkaloid composition of B. cusia

Protectin conjugates in tissue regeneration 1 alleviates sepsis-induced acute lung injury by inhibiting ferroptosis

Background: Acute lung injury (ALI) is a common and serious complication of sepsis with high mortality. Ferroptosis, categorized as programmed cell death, contributes to the development of lung injury. Protectin conjugates in tissue regeneration 1 (PCTR1) is an endogenous lipid mediator that exerts protective effects against multiorgan injury. However, the role of PCTR1 in the ferroptosis of sepsis-related ALI remains unknown. Methods: A pulmonary epithelial cell line and a mouse model of ALI stimulated with lipopolysaccharide (LPS) were established in vitro and in vivo. Ferroptosis biomarkers, including ferrous (Fe2+), glutathione (GSH), malondialdehyde (MDA) and 4-Hydroxynonenal (4-HNE), were assessed by relevant assay kits. Glutathione peroxidase 4 (GPX4) and prostaglandin-endoperoxide synthase 2 (PTGS2) protein levels were determined by western blotting. Lipid peroxides were examined by fluorescence microscopy and flow cytometry. Cell viability was determined by a CCK-8 assay kit. The ultrastructure of mitochondria was observed with transmission electron microscopy. Morphology and inflammatory cytokine levels predicted the severity of lung injury. Afterward, related inhibitors were used to explore the potential mechanism by which PCTR1 regulates ferroptosis. Results: PCTR1 treatment protected mice from LPS-induced lung injury, which was consistent with the effect of the ferroptosis inhibitor ferrostatin-1. PCTR1 treatment decreased Fe2+, PTGS2 and lipid reactive oxygen species (ROS) contents, increased GSH and GPX4 levels and ameliorated mitochondrial ultrastructural injury. Administration of LPS or the ferroptosis agonist RSL3 resulted in reduced cell viability, which was rescued by PCTR1. Mechanistically, inhibition of the PCTR1 receptor lipoxin A4 (ALX), protein kinase A (PKA) and transcription factor cAMP-response element binding protein (CREB) partly decreased PCTR1 upregulated GPX4 expression and a CREB inhibitor blocked the effects ofPCTR1 on ferroptosis inhibition and lung protection. Conclusion: This study suggests that PCTR1 suppresses LPS-induced ferroptosis via the ALX/PKA/CREB signaling pathway, which may offer promising therapeutic prospects in sepsis-related ALI

Quantitative evaluation of structural complexity in Shengquan minefield based on improved local linear embedding algorithm

Mine water disaster is closely related to the geological structure of the mine. A scientific evaluation of the complexity of the minefield structure can greatly contribute to mining safety. In this study, we optimized the fault impact index in the study area and proposed an improved local linear embedding (LLE) algorithm by investigating the Shengquan coal mine. We analyzed the fault formation age and connectivity between faults in the study area, combined with topology theory, and based on previous studies. The mechanical properties of faults and the influence of small faults on the strata can be combined to derive a quantitative evaluation model of the structural complexity of mines. Based on the evaluation model, the study area was divided into a simple structural area, a medium structural area, and a complex structural area. By comparing the location of water inrush points in recent years with the three-dimensional high-density electrical exploration of the 21,304 working face, the effectiveness and rationality of structural complexity zoning were determined

Computational Efficient Motion Planning Method for Automated Vehicles Considering Dynamic Obstacle Avoidance and Traffic Interaction.

In complex driving scenarios, automated vehicles should behave reasonably and respond adaptively with high computational efficiency. In this paper, a computational efficient motion planning method is proposed, which considers traffic interaction and accelerates calculation. Firstly, the behavior is decided by connecting the points on the unequally divided road segments and lane centerlines, which simplifies the decision-making process in both space and time span. Secondly, as the dynamic vehicle model with changeable longitudinal velocity is considered in the trajectory generation module, the C/GMRES algorithm is used to accelerate the calculation of trajectory generation and realize on-line solving in nonlinear model predictive control. Meanwhile, the motion of other traffic participants is more accurately predicted based on the driver's intention and kinematics vehicle model, which enables the host vehicle to obtain a more reasonable behavior and trajectory. The simulation results verify the effectiveness of the proposed method

Predicting Dynamics of the Potential Breeding Habitat of Larus saundersi by MaxEnt Model under Changing Land-Use Conditions in Wetland Nature Reserve of Liaohe Estuary, China

Identifying waterfowl habitat suitability under changing environments, especially land-use change, is crucial to make waterfowl habitat conservation planning. We took Wetland Nature Reserve of Liaohe Estuary, the largest breeding area of Saunders’s Gulls (Larus saundersi) in the world, as our study area, generated land-use-type maps through interpretation of satellite images from four different years (1988, 2000, 2009, 2017), and predicted the potential breeding habitat of Saunders’s Gulls by MaxEnt model based on the land-use map, along with other environmental variables (NDVI, distance to roads and artificial facilities, distance to rivers and water bodies, DEM and distance to shoreline) for the four years, respectively. The models were evaluated using the area under the curve (AUC). We analyzed the changes of the breeding habitat from 1988 to 2017 and utilized RDA to explore the relationships among the changes of suitable habitat of Larus saundersi and the dynamics of land uses. Our results showed that the most suitable habitat decreased by 1286.46 ha during 1988-2009 and increased by 363.51 ha from 2009 to 2017. The suitable habitat decreased by 582.48 ha from 1988 to 2009 and then increased to 1848.96 ha in 2017, while the unsuitable habitat increased by 2793.87 ha during 1988–2009 and then decreased by 178.83 ha from 2009 to 2017. We also found that land use, distance to the coastline, distance to artificial facilities, distance to rivers, distance to roads, and NDVI had certain degrees of impact on the Larus saundersi distribution. The contribution of land use ranged from 16.4% to 40.3%, distance to coastline from 34.7% to 48.0%, distance to artificial facilities from 5.9% to 11.1%, distance to rivers from 5.5% to 11.0%, distance to roads from 3.9% to 12.5%, and NDVI from 0.3% to 6.3%. The change in suitable habitat of Larus saundersi has a positive relationship with the change of seepweed marsh. Human-induced changes in seepweed marsh and coastline position are the main factors influencing the potential breeding habitat of Saunders’s Gulls. We suggest strict conservation of seepweed marsh and implementation of habitat management practices to better protect Saunders’ Gull’s breeding habitat

GLT-T++: Global-Local Transformer for 3D Siamese Tracking with Ranking Loss

Siamese trackers based on 3D region proposal network (RPN) have shown remarkable success with deep Hough voting. However, using a single seed point feature as the cue for voting fails to produce high-quality 3D proposals. Additionally, the equal treatment of seed points in the voting process, regardless of their significance, exacerbates this limitation. To address these challenges, we propose a novel transformer-based voting scheme to generate better proposals. Specifically, a global-local transformer (GLT) module is devised to integrate object- and patch-aware geometric priors into seed point features, resulting in robust and accurate cues for offset learning of seed points. To train the GLT module, we introduce an importance prediction branch that learns the potential importance weights of seed points as a training constraint. Incorporating this transformer-based voting scheme into 3D RPN, a novel Siamese method dubbed GLT-T is developed for 3D single object tracking on point clouds. Moreover, we identify that the highest-scored proposal in the Siamese paradigm may not be the most accurate proposal, which limits tracking performance. Towards this concern, we approach the binary score prediction task as a ranking problem, and design a target-aware ranking loss and a localization-aware ranking loss to produce accurate ranking of proposals. With the ranking losses, we further present GLT-T++, an enhanced version of GLT-T. Extensive experiments on multiple benchmarks demonstrate that our GLT-T and GLT-T++ outperform state-of-the-art methods in terms of tracking accuracy while maintaining a real-time inference speed. The source code will be made available at https://github.com/haooozi/GLT-T.Comment: Need further revisio

DIC imaging for identification of motor and sensory nerves

Identification of motor and sensory nerves is important in applications such as nerve injury repair. Conventional practice relies on time consuming staining methods for this purpose. Here, we use laser scanning infrared differential interference contrast (IR-DIC) microscopy for label-free observation of the two types of nerve. Ventral and dorsal nerve roots of adult beagle dogs were collected and sections of different thicknesses were imaged with an IR-DIC microscope. Different texture patterns of the IR-DIC images of the motor and sensory nerve can be distinguished when the section thickness increases to 40μm. This suggests that nerve fibers in motor and sensory nerves have different distribution patterns. The result hints a potential new way for more rapid identification of nerve type in peripheral nerve repair surgery