Feature-based molecular networking with MS2LDA to profile compounds in Lanbuzheng based on ultra-high-performance liquid chromatography-quadrupole Exactive Orbitrap high-resolution mass spectrometry.

Lanbuzheng (Geum japonicum Thunb. var. chinense Bolle), a plant found in Southwest China, is a traditional Chinese medicine that promotes hematopoiesis and antioxidant functions.

Catecholamines-based myocardial injury after acute ischemic stroke and effects of Naoxintong capsule therapy.

Background: Myocardial injury (MI) after acute ischemic stroke (AIS) poses a significant threat to patient prognosis. However, effective intervention strategies are currently lacking.

Purpose: To elucidate the mechanism of MI after AIS and effects of Naoxintong capsule (NXT) therapy.

Efficient and markerless gene integration with SlugCas9-HF in Kluyveromyces marxianus.

The nonconventional yeast Kluyveromyces marxianus has potential for industrial production, but the lack of advanced synthetic biology tools for precise engineering hinders its rapid development. Here, we introduce a CRISPR-Cas9-mediated multilocus integration method for assembling multiple exogenous genes. Using SlugCas9-HF, a high-fidelity Cas9 nuclease, we enhance gene editing precision.

Engineering of a high-fidelity Cas12a nuclease variant capable of allele-specific editing.

CRISPR-Cas12a, often regarded as a precise genome editor, still requires improvements in specificity. In this study, we used a GFP-activation assay to screen 14 new Cas12a nucleases for mammalian genome editing, successfully identifying 9 active ones. Notably, these Cas12a nucleases prefer pyrimidine-rich PAMs.

Phage-assisted evolution of compact Cas9 variants targeting a simple NNG PAM.

Compact Cas9 nucleases hold great promise for therapeutic applications. Although several compact Cas9 nucleases have been developed, many genomic loci still could not be edited due to a lack of protospacer adjacent motifs (PAMs). We previously developed a compact SlugCas9 recognizing an NNGG PAM.

Skeletal muscle-specific inducible AMPKα1/α2 knockout mice develop muscle weakness, glycogen depletion, and fibrosis that persists during disuse atrophy.

The 5' adenosine monophosphate-activated protein kinase (AMPK) is an important skeletal muscle regulator implicated as a possible therapeutic target to ameliorate the local undesired deconditioning of disuse atrophy. However, the muscle-specific role of AMPK in regulating muscle function, fibrosis, and transcriptional reprogramming during physical disuse is unknown. The purpose of this study was to determine how the absence of both catalytic subunits of AMPK in skeletal muscle influences muscle force production, collagen deposition, and the transcriptional landscape.

In Vivo Base Editing of Rescues Type 3 Long QT Syndrome in Mice.

Background: Pathogenic variants in can result in long QT syndrome type 3, a life-threatening genetic disease. Adenine base editors can convert targeted A T base pairs to G C base pairs, offering a promising tool to correct pathogenic variants.

Methods: We generated a long QT syndrome type 3 mouse model by introducing the T1307M pathogenic variant into the gene.

Effects of nitrogen inputs and mowing on the abundance and species richness of herbivorous insects in a meadow steppe.

To reveal the effects of nitrogen (N) inputs and mowing on the abundance and richness of insect community in meadow steppe, we investigated the abundance and diversity of herbivorous insects under four treatments in Inner Mongolia meadow steppe in August 2022, including control, N addition, mowing, and combined N addition and mowing. At a long-term control experimental platform, we collected insects using the vacuum sampling method. The results showed that N addition significantly increased the abundance of herbivorous insects, and mowing significantly decreased the abundance of herbivorous insects.

A highly specific CRISPR-Cas12j nuclease enables allele-specific genome editing.

The CRISPR-Cas system can treat autosomal dominant diseases by nonhomologous end joining (NHEJ) gene disruption of mutant alleles. However, many single-nucleotide mutations cannot be discriminated from wild-type alleles by current CRISPR-Cas systems. Here, we functionally screened six Cas12j nucleases and determined Cas12j-8 as an ideal genome editor with a hypercompact size.

Genome editing with natural and engineered CjCas9 orthologs.

CjCas9 is one of the smallest CRISPR-associated (Cas9) nucleases for mammalian genome editing. However, it requires a long NRYAC (R = A or G; Y = C or T) protospacer-adjacent motif (PAM), limiting its DNA targeting scope. In this study, we investigated the PAMs of three CjCas9 orthologs, including Hsp1Cas9, Hsp2Cas9, and CcuCas9, by performing a GFP-activation assay.

Closely related type II-C Cas9 orthologs recognize diverse PAMs.

The RNA-guided CRISPR/Cas9 system is a powerful tool for genome editing, but its targeting scope is limited by the protospacer-adjacent motif (PAM). To expand the target scope, it is crucial to develop a CRISPR toolbox capable of recognizing multiple PAMs. Here, using a GFP-activation assay, we tested the activities of 29 type II-C orthologs closely related to Nme1Cas9, 25 of which are active in human cells.

Integrated Strategy From , , to for Predicting Active Constituents and Exploring Molecular Mechanisms of Tongfengding Capsule for Treating Gout by Inhibiting Inflammatory Responses.

The study of screening active constituents from traditional Chinese medicine (TCM) is important for explicating the mechanism of action of TCM and further evaluating the safety and efficacy effectively. However, detecting and identifying the active constituents from complicated biological samples still remain a challenge. Here, a practical, quick, and novel integrated strategy from , , to for rapidly screening the active constituents was developed.

Effect of CYP3A4 Inhibitors and Inducers on Pharmacokinetics and Pharmacodynamics of Saxagliptin and Active Metabolite M2 in Humans Using Physiological-Based Pharmacokinetic Combined DPP-4 Occupancy.

We aimed to develop a physiological-based pharmacokinetic and dipepidyl peptidase 4 (DPP-4) occupancy model (PBPK-DO) characterized by two simultaneous simulations to predict pharmacokinetic (PK) and pharmacodynamic changes of saxagliptin and metabolite M2 in humans when coadministered with CYP3A4 inhibitors or inducers. Ketoconazole, delavirdine, and rifampicin were selected as a CYP3A4 competitive inhibitor, a time-dependent inhibitor, and an inducer, respectively. Here, we have successfully simulated PK profiles and DPP-4 occupancy profiles of saxagliptin in humans using the PBPK-DO model.

Proteomic profiling sheds light on alkali tolerance of common wheat (Triticum aestivum L.).

Alkali (high-pH) stress is an important factor limiting agricultural production and has complex effects on plant metabolism. Transcriptomics is widely used in the discovery of stress-response genes, but it provides only a rough estimation for gene expression. Proteomics may be more helpful than transcriptomics for the discovery and identification of stress-response genes.