As a core component of power transmission, substations often receive little attention regarding their internal ecological conditions. In this study, we established artificial grasslands at the Xining 330 kV Huayuan Substation and evaluated their ecological benefits. Our results showed that Duchesnea indica (SP1) and Poa annua (SP4) exhibited extremely high coverage (>90%) and aboveground biomass, positioning them as ideal pioneer species for vegetation restoration. The establishment of artificial grasslands significantly increased soil available nutrient content and moisture, reduced soil pH, and created a more favorable microenvironment for microbial communities. Significant differences were observed in the β-diversity of soil bacteria, fungi, and protists: bacterial communities diverged only between the artificial grasslands and the control, whereas fungal and protist communities not only differed from the control but also showed significant separation among different grass species treatments. Atificial grasslands substantially enhanced the complexity of the multi-domain network encompassing bacteria, fungi, and protists; importantly, the degree centrality of the three groups did not differ, underscoring their equivalent importance in maintaining network structure. Untargeted LC-MS metabolomics identified a total of 877 soil metabolites, with lipids and lipid-like molecules being the predominant class. PLS-DA analysis revealed significant differences in metabolites among treatments, and random forest analysis identified protists and their associated micro-food webs as the primary factors driving variation in metabolite composition. In summary, the construction of artificial grasslands within substations triggered multidimensional and profound ecosystem changes, driving cascading effects that extended from aboveground plant communities through the soil micro-food web to soil metabolites. These findings provide a scientific basis for developing ecological restoration technologies based on targeted regulation of the soil micro-food web, and advance our understanding of multi-trophic interactions in terrestrial ecosystems.

Rheum tanguticum is renowned for its medicinal properties, including purgative, anti-inflammatory and hepatoprotective effects, primarily attributed to anthraquinones (AQs). However, the molecular mechanisms of AQs biosynthesis have largely been hindered by insufficient genomic resources and functional genomics investigations. Here, we employed multi-omics approaches to address this knowledge gap. The high-quality T2T-level genome was constructed with a size of 2.68 Gb and a contig N50 of 233.65 Mb. Functional annotation revealed that the specific and expanded gene families of R. tanguticum are involved in efficient energy metabolism and secondary metabolite biosynthesis, providing a molecular basis for its stress adaptation and medicinal value. Integrated widely targeted metabolomics, spatial metabolomics and targeted quantification of AQs, we successfully elucidate the AQs spatiotemporal accumulation patterns. Seeds and leaves are key sites for the synthesis and transport of free AQs, while the root core serves as the primary location for the synthesis and accumulation of conjugated AQs. Through integrated genomic, transcriptomic, metabolomic and functional validation analyses, we preliminarily characterised the positive regulatory role of RtPKSIII-8 in the synthesis of aloe-emodin and emodin, as well as the potential function of RtUGT85AD11 in the biosynthesis of AQs and flavonoids. These findings provide essential genomic and functional data for deciphering AQs biosynthetic pathways and lay a theoretical foundation for the medicinal development and genetic improvement of R. tanguticum.

ABSTRACT Rational design of molecular glues (MGs) remains challenging, as most have been discovered serendipitously and have found limited application in antivirals. Previously, we identified the enteroviral 3A protein as a viral suppressor of RNAi (VSR) that functions through homodimerization to inhibit the antiviral RNA interference (RNAi) pathway. Herein, capitalizing on this homodimerization mechanism, we rationally designed 3A‐targeting broad‐spectrum anti‐enteroviral molecular glues targeting the dimeric interface to induce dysfunctional dimerization. The optimal compound, VTP‐32, exhibited good binding affinity with 3A (KD = 0.29 µm), potent and pan‐enterovirus (groups A, B, D) antiviral effects (EC50 = 0.21–0.92 µm), and good safety (CC50 > 500 µm). VTP‐32 treatment (20 mg/kg) could effectively reduce viral load, alleviate clinical symptoms, and improve survival in EV‐A71‐infected mouse models. Mechanistic studies revealed that VTP‐32 stabilizes 3A protein into an abnormal dimer, promotes viral siRNA generation, and ultimately leads to RNAi‐mediated viral genome degradation. Overall, this study provides a promising countermeasure against enteroviral diseases and a rational design strategy for developing antiviral molecular glues.

Rheum tanguticum (R. tanguticum) originates from high-altitude regions such as Qinghai and Gansu in China. It serves as both a precious traditional medicinal botanical drug and a potential functional food, and its extensive pharmacological activities have stimulated global demand. However, existing research predominantly focuses on optimizing individual drying techniques or single bioactive metabolites, failing to integrate multi-omics technologies to elucidate the chemical and physical alterations induced by drying. To this end, we evaluated five drying methods by integrating an approach that links physical indicators with chemical composition, revealing that, for the preservation of physical indicators, especially color and rehydration properties, vacuum freeze drying (LD) is optimal; for the retention of the key pharmaceutical metabolite free anthraquinones, microwave drying (WB) is the most effective. Whereas in the comprehensive evaluation of multi-dimensional quality, SG is optimal. Metabolomic analysis confirmed that LD and YG were most effective in preserving key metabolites. Flavonoids, phenolic acids, and amino acids collectively form the core metabolites of differential metabolites in dried R. tanguticum, primarily enriched in pathways such as purine metabolism. Collectively, this work systematically elucidates the impact of drying on R. tanguticum’s metabolite profile and physicochemical traits, offering both a theoretical basis for its precision processing and a transferable framework for optimizing drying techniques in related medicinal edible plants.

Gastrodia elata is a traditional and valuable Chinese medicinal herb that possesses both medicinal and edible properties; its clinical efficacy is significantly influenced by its quality. However, as cultivation expands, the quality of G. elata from different producing areas exhibits considerable variability. Therefore, a systematic evaluation of G. elata from different origins is essential for cultivating high-quality medicinal materials. This study innovatively integrates multi-source active ingredient data with environmental factors and employs a machine learning-SHAP analysis framework to explore the above scientific issues, thereby achieving precise quantification of factor contributions and spatial prediction of quality. The results indicate that the quality of wild G. elata surpasses that of cultivated varieties, with significantly higher levels of gastrodin, 4-Hydroxybenzyl alcohol, parishin A, and parishin B. Notably, 96% of cultivated G. elata meet the basic requirements outlined in the Chinese Pharmacopoeia, although the contents of effective components vary significantly across regions. Among these, 4-Hydroxybenzyl alcohol emerges as the most significant effective component for distinguishing G. elata from different origins, followed closely by gastrodin. Spatial distribution maps reveal that regions with high concentrations of gastrodin and 4-Hydroxybenzyl alcohol are predominantly located in the traditionally interconnected areas of eastern Sichuan, northeastern Yunnan, and western Guizhou. Furthermore, SHAP analysis identifies the mean temperature of the coldest quarter as the most critical determinant affecting its quality. This study not only confirms traditional knowledge but also uncovers new patterns, thereby providing important scientific evidence and data support for the standardized cultivation and systematic quality evaluation of G. elata.

Anisodus tanguticus, a rare medicinal plant producing clinically vital tropane alkaloids: anisodine, anisodamine, scopolamine, and atropine, faces challenges in sustainable cultivation due to wild resource scarcity and suboptimal farming practices. While P addition is known to influence secondary metabolites, its microbiome mediated effects on alkaloid synthesis remain unclear. Therefore, in this study, we systematically analysed the mechanism of P on the association between alkaloid content of Anisodus tanguticus and microbiome by setting P addition gradients (LP: 120 kg/hm2, MP: 180 kg/hm2, HP: 240 kg/hm2). The results demonstrated that P addition significantly reduced the content of 4 alkaloids in roots, stems and leaves. While microbial alpha-diversity remained unresponsive, but induced a structural reorganization of the microbial community, encompassing rhizosphere and bulk soil bacteria, which exhibited a marked association with alkaloids, intensified nutrient competition through the nitrogen cycling and organic matter catabolism pathways. Concurrently, the abundance of pathogenic functions within the fungal community underwent a significant increase with P addition. In this study, it was demonstrated that the addition of P inhibits alkaloid accumulation through dual pathways: (1) soil C:N:P stoichiometry imbalance interferes with plant secondary metabolism resource allocation; and (2) the introduction of pathogenic fungi hinders alkaloid synthesis and accumulation in plants. These findings provide a theoretical basis for the cultivation of Anisodus tanguticus by reducing the amount and increasing the efficiency of chemical fertilisers. The study suggests that precise fertilisation of medicinal plants needs to take into account the balance of soil nutrients and microbial function regulation. By suppressing the pathogenic bacterial flora and optimising the abundance of soil probiotic bacteria, it is possible to reduce the amount of phosphorus fertiliser applied and at the same time increase the yield of medicinal active ingredients, which is of great practical value in achieving the sustainable use of ethnomedicinal resources.

R. tanguticum (Rheum tanguticum Maxim. ex Regel) is a herbaceous plant belonging to Polygonaceae family and Rheum L. genus. It holds considerable value in culinary and medicinal realms, primarily due to their rich Anthraquinones (AQs) content. Understanding the molecular mechanisms that regulate AQs biosynthesis is a prerequisite for increasing their yield. MYB transcription factors (TFs) can regulate the synthesis of a variety of plant secondary metabolites. However, only a few research have explored the role of MYB TFs in Rheum L. species. In this study, 1054 MYB genes from four Rheum L. species were identified. The number of MYB genes in each species was similar, distributed across 11 chromosomes. To investigate the phylogeny of identified MYB TFs, they were classified into four subfamilies. Sequence characteristics, phylogenetic relationships, evolutionary trends, and tissue expression of MYB genes in Rheum L. species were further studied. Subsequently, 12 MYB genes were selected, which shown differential expression in different tissues. Further research on these genes indicated a significant correlation with genes in shikimate pathway and polyketide pathway of AQs biosynthesis. Protein-protein interaction simulations in Arabidopsis thaliana and qRT-PCR experiments further confirmed this situation. This research lays the foundation for studying molecular mechanisms by which MYB TFs regulates AQs biosynthesis in four Rheum L. species.

The extraordinary chemodiversity of secondary metabolites in Rheum tanguticum Maxim. ex Balf. underpins its promise as a great Food-Medicine Homology crop. Elucidating how these metabolites accumulate and are spatially organized during root maturation is therefore an essential prerequisite for quality-oriented cultivation and processing.Here, we harvested roots at three developmental stages and integrated MALDI-MSI, UPLC-ESI-MS/MS and RNA-seq to dissect the morphological, metabolic and transcriptional dynamics governing quality formation. Quantitative and imaging results indicate that AQs and flavonoids account for a significant proportion of the bioactive constituents in rhubarb roots and exhibit complementary spatial gradients that expand centripetally with root age. Transcriptome co-mapping identified stage-specific PKSIII and MYBs expression hotspots that precisely colocalize with these metabolite maxima, providing the first genetic evidence for their coordinated biosynthesis in R. tanguticum.These findings not only redefine the spatiotemporal blueprint of metabolite accumulation in rhubarb roots, but also offer high-resolution targets for marker-assisted breeding and targeted processing strategies to maximize both medicinal efficacy and functional-food value.

Co-inhibition of histone deacetylase (HDAC) and cyclin-dependent kinase (CDK) synergizes to produce enhanced antitumor effects and potentially overcomes the drug resistance. In this work, we discovered a series of novel CDK9/HDACs dual inhibitors. Among them, compound 8e was identified to show potent CDK9 and HDAC1 inhibitory activities, with IC50 values at 88.4 and 168.9 nM, respectively, and exhibited antiproliferative capacities against hematological and solid tumor cells. Meanwhile, 8e showed high selectivity for CDK9 and HDAC1, remarkably induced MV-4-11 cell apoptosis and S cell cycle arrests. Furthermore, 8e possessed a significant antitumor potency with a T/C value of 29.98% in the MV-4-11 xenograft model. Interestingly, a potent FLT3/HDAC dual inhibitor 9e was also identified (FLT3/HDAC1/3 IC50 = 30.4/52.4/14.7 nM) and found to possess powerful apoptosis induction ability in MV-4-11 cell and potent antiproliferative capacities against FLT3 mutant-transformed BaF3 cells. Overall, our work provided valuable lead compounds for dual inhibitors with potent anticancer activity.

Acute myeloid leukaemia(AML)is characterized mainly by an increase in the number of myeloid cells in the bone marrow and a decrease in the number of mature cells;AML accounts for 28％of leukaemia cases,and it has a five-year survival rate of only 30.5％[1].

The outcomes of FLT3-ITD acute myeloid leukaemia (AML) have been improved since the approval of FLT3 inhibitors (FLT3i). However, approximately 30-50% of patients exhibit primary resistance (PR) to FLT3i with poorly defined mechanisms, posing a pressing clinical unmet need. Here, we identify C/EBPα activation as a top PR feature by analyzing data from primary AML patient samples in Vizome. C/EBPα activation limit FLT3i efficacy, while its inactivation synergistically enhances FLT3i action in cellular and female animal models. We then perform an in silico screen and identify that guanfacine, an antihypertensive medication, mimics C/EBPα inactivation. Furthermore, guanfacine exerts a synergistic effect with FLT3i in vitro and in vivo. Finally, we ascertain the role of C/EBPα activation in PR in an independent cohort of FLT3-ITD patients. These findings highlight C/EBPα activation as a targetable PR mechanism and support clinical studies aimed at testing the combination of guanfacine with FLT3i in overcoming PR and enhancing the efficacy of FLT3i therapy.

Current therapy for acute myeloid leukemia (AML) is largely hindered by the development of drug resistance of commonly used chemotherapy drugs, including cytarabine, daunorubicin, and idarubicin. In this study, we investigated the molecular mechanisms underlying the chemotherapy drug resistance and potential strategy to improve the efficacy of these drugs against AML. By analyzing data from ex vivo drug-response and multi-omics profiling public data for AML, we identified autophagy activation as a potential target in chemotherapy-resistant patients. In THP-1 and MV-4-11 cell lines, knockdown of autophagy-regulated genes ATG5 or MAP1LC3B significantly enhanced AML cell sensitivity to the chemotherapy drugs cytarabine, daunorubicin, and idarubicin. In silico screening, we found that chloroquine phosphate mimicked autophagy inactivation. We showed that chloroquine phosphate dose-dependently down-regulated the autophagy pathway in MV-4-11 cells. Furthermore, chloroquine phosphate exerted a synergistic antitumor effect with the chemotherapy drugs in vitro and in vivo. These results highlight autophagy activation as a drug resistance mechanism and the combination therapy of chloroquine phosphate and chemotherapy drugs can enhance anti-AML efficacy.

Abstract Two new maytansinoids, N-methyltreflorine (1) and methyltrewiasine (2), were isolated from the dried fruits of Trewia nudiflora, together with three known congeners (3 – 5). Their structures were elucidated by spectroscopic methods, and the absolute configuration of 1 and 2 was determined by X-ray crystallographic analysis. Compounds 1 – 5 exhibited strong cytotoxicity against human tumor cell lines, including HeLa, MV-4 – 11, and MCF-7, with IC50 values ranging from 0.12 to 11 nM. Compounds 1 and 4 also showed inhibitory activity against the MCF-7/ADR cell line with IC50 values of 13 and 28 nM, respectively. Compounds 1 and 2 significantly inhibited tubulin polymerization in vitro with IC50 values of 3.6 and 3.2 µM, respectively.

Three maytansinoids with strong cytotoxicities, dehydrotrewiasine, maytanbutine, and trewiasine, were isolated and identified from Trewia nudiflora, and maytanbutine was obtained from this plant for the first time. A quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction combined with high-performance liquid chromatography (HPLC) was established to determine the three maytansinoids in T. nudiflora. The effects of major factors on the extraction efficiency of the QuEChERS method were evaluated and the optimal conditions using acetonitrile-ethyl acetate (1:1, v/v) as the extraction solvent and PestiCarb as the clean-up sorbents were established. Compared with Soxhlet extraction (SE) and ultrasonic-assisted extraction (UAE), the QuEChERS method was easy-to-operate and afforded a cleaner extract. A phenomenex HyperClone BDS C-18 column was used for HPLC analysis. Methanol-acetonitrile-water was chosen as mobile phase for gradient elution. Method validation showed that all analytes showed good linearity (r > 0.999) over the investigated ranges and satisfactory recoveries ranging from 95.0% to 105.0%. The developed QuEChERS-HPLC method was simple, efficient, and applicable to the determination of maytansinoids in T. nudiflora. (C) 2021 Elsevier B.V. All rights reserved.

Checkpoint kinase 1 inhibitors (CHK1i) have shown impressive single-agent efficacy in treatment of certain tumors, as monotherapy or potentiators of chemotherapy in clinical trials, but the sensitive tumor types and downstream effectors to dictate the therapeutic responses to CHK1i remains unclear. In this study we first analyzed GDSC (Genomics of Drug Sensitivity in Cancer) and DepMap database and disclosed that hematologic malignancies (HMs) were relatively sensitive to CHK1i or CHK1 knockdown. This notion was confirmed by examining PY34, a new and potent in-house selective CHK1i, which exhibited potent anti-HM effect in vitro and in vivo, as single agent. We demonstrated that the downregulation of c-Myc and its signaling pathway was the common transcriptomic profiling response of sensitive HM cell lines to PY34, whereas overexpressing c-Myc could partially rescue the anticancer effect of PY34. Strikingly, we revealed the significant correlations between downregulation of c-Myc and cell sensitivity to PY34 in 17 HM cell lines and 39 patient-derived cell (PDC) samples. Thus, our results demonstrate that HMs are more sensitive to CHK1i than solid tumors, and c-Myc downregulation could represent the CHK1i efficacy in HMs.

Diffuse large B-cell lymphoma (DLBCL) is the most widespread type of non-Hodgkin lymphoma (NHL). As the most aggressive form of the DLBCL, the activated B-cell-like (ABC) subtype is often resistant to standard chemotherapies. Bruton’s tyrosine kinase (BTK) inhibitor ibrutinib provides a potential therapeutic approach for the DLBCL but fails to improve the outcome in the phase III trial. In the current study, we investigated the molecular mechanisms underlying ibrutinib resistance and explored new combination therapy with ibrutinib. We generated an ibrutinib-resistant ABC-DLBCL cell line (OCI-ly10-IR) through continuous exposure to ibrutinib. Transcriptome analysis of the parental and ibrutinib-resistant cell lines revealed that the ibrutinib-resistant cells had significantly lower expression of the unfolded protein response (UPR) marker genes. Overexpression of one UPR branch-XBP1s greatly potentiated ibrutinib-induced apoptosis in both sensitive and resistant cells. The UPR inhibitor tauroursodeoxycholic acid (TUDCA) partially reduced the apoptotic rate induced by the ibrutinib in sensitive cells. The UPR activator 2-deoxy- D -glucose (2-DG) in combination with the ibrutinib triggered even greater cell growth inhibition, apoptosis, and stronger calcium (Ca 2+ ) flux inhibition than either of the agents alone. A combination treatment of ibrutinib (15 mg·kg −1 ·d −1 , po.) and 2-DG (500 mg/kg, po, b.i.d.) synergistically retarded tumor growth in NOD/SCID mice bearing OCI-ly10-IR xenograft. In addition, ibrutinib induced the UPR in the sensitive cell lines but not in the resistant cell lines of the DLBCL. There was also a combined synergistic effect in the primary resistant DLBCL cell lines. Overall, our results suggest that targeting the UPR could be a potential combination strategy to overcome ibrutinib resistance in the DLBCL.

Tetrandrine (Tet) bisbenzylisoquinoline alkaloids isolated from Stephania tetrandra and other related species of Menispermaceae. It has been demonstrated to have positive therapeutic effects on cardiovascular disease, hypertension, silicosis, autoimmune diseases. In recent years, some reports have shown that Tet has anticancer activity in human cancers. To explore the pharmacological activity and mechanism of Tet on colon cancer and its unique advantages as a natural product. In the present study, analyses of the cell cycle, apoptosis, targets prediction, molecular docking, and alterations in protein levels were performed to elucidate how Tet functions in colon cancer. We found that Tet robustly induced arrest at the G1 phase in colon cancer cell line HT-29. It induced HT-29 cell apoptosis in a dose-dependent manner. Similarly, analysis of protein expression levels in HT-29 cells showed down-regulation of Bcl-2, pro-caspase 3, pro-caspase 8, PARP, cyclin D1 (CCND1), cyclin-dependent kinase 4 (CDK 4), and up-regulation of Bax, active caspase 3, and active caspase 8. These results indicate that Tet induces apoptosis of colon cancer cells through the mitochondrial pathway and caspase family pathway. Molecular docking showed interaction effects and binding energy. Comparing with the CDK4 inhibitors ribociclib and palbociclib, the docking energy is similar to the docked amino acid residues. Therefore, we conclude that Tet and the CCND1/CDK4 compound could form hydrogen bonds and a stable compound structure, which can inhibit colon cancer cells proliferation by regulating CCND1/CDK4 compound and its downstream proteins phosphorylated Rb (p-Rb). In summary, Tet may be a potential drug for colon cancer therapy.

Shikonin, a natural naphthoquinone compound derived from the herb Lithospermum erythrorhizon, is widely used for its various pharmacological activities. However, its potential interactions with other medications by inhibiting human carboxylesterases 2 (hCE2) remain unknown. In this study, the inhibitory effects of shikonin on the activity of hCE2 in human liver microsomes are investigated by using fluorescein diacetate (FD), N-(2-butyl-1,3-dioxo-2,3-dihydro-1H-phenalen-6-yl)-2-chloroacetamide (NCEN), and CPT-11 as substrates of hCE2. The results demonstrate that shikonin significantly inhibits the activity of hCE2 when FD and NCEN are used as substrates, whereas the half inhibition concentration value of shikonin increased by 5-30 times when CPT-11 was used as the substrate. The inhibition types of shikonin against hCE2 activity reflected by 3 substrates were all best fit to noncompetitive manners. In addition, shikonin was found to distinctly suppress endogenous hCE2 activity, characterized with attenuated fluorescence. Furthermore, for drugs metabolized by hCE2 with the similar binding sites with FD or NCEN, the estimated magnitudes of area under the curve variation were approximately 9-357% in the presence of shikonin. Also, the area under the curve of CPT-11 could be increased by 1-14% following administration of shikonin. These findings have clear clinical implications for the combination of shikonin and hCE2-metabolizing prodrugs.