Mosaic sarbecovirus nanoparticles elicit cross-reactive responses in pre-vaccinated animals.

Immunization with mosaic-8b (nanoparticles presenting 8 SARS-like betacoronavirus [sarbecovirus] receptor-binding domains [RBDs]) elicits more broadly cross-reactive antibodies than homotypic SARS-CoV-2 RBD-only nanoparticles and protects against sarbecoviruses. To investigate original antigenic sin (OAS) effects on mosaic-8b efficacy, we evaluated the effects of prior COVID-19 vaccinations in non-human primates and mice on anti-sarbecovirus responses elicited by mosaic-8b, admix-8b (8 homotypics), or homotypic SARS-CoV-2 immunizations, finding the greatest cross-reactivity for mosaic-8b. As demonstrated by molecular fate mapping, in which antibodies from specific cohorts of B cells are differentially detected, B cells primed by WA1 spike mRNA-LNP dominated antibody responses after RBD-nanoparticle boosting. While mosaic-8b- and homotypic-nanoparticles boosted cross-reactive antibodies, de novo antibodies were predominantly induced by mosaic-8b, and these were specific for variant RBDs with increased identity to RBDs on mosaic-8b. These results inform OAS mechanisms and support using mosaic-8b to protect COVID-19-vaccinated/infected humans against as-yet-unknown SARS-CoV-2 variants and animal sarbecoviruses with human spillover potential.

Engineering RNA export for measurement and manipulation of living cells.

A system for programmable export of RNA molecules from living cells would enable both non-destructive monitoring of cell dynamics and engineering of cells capable of delivering executable RNA programs to other cells. We developed genetically encoded cellular RNA exporters, inspired by viruses, that efficiently package and secrete cargo RNA molecules from mammalian cells within protective nanoparticles. Exporting and sequencing RNA barcodes enabled non-destructive monitoring of cell population dynamics with clonal resolution. Further, by incorporating fusogens into the nanoparticles, we demonstrated the delivery, expression, and functional activity of exported mRNA in recipient cells. We term these systems COURIER (controlled output and uptake of RNA for interrogation, expression, and regulation). COURIER enables measurement of cell dynamics and establishes a foundation for hybrid cell and gene therapies based on cell-to-cell delivery of RNA.

Neutralizing monoclonal antibodies elicited by mosaic RBD nanoparticles bind conserved sarbecovirus epitopes.

Increased immune evasion by SARS-CoV-2 variants of concern highlights the need for new therapeutic neutralizing antibodies. Immunization with nanoparticles co-displaying spike receptor-binding domains (RBDs) from eight sarbecoviruses (mosaic-8 RBD-nanoparticles) efficiently elicits cross-reactive polyclonal antibodies against conserved sarbecovirus RBD epitopes. Here, we identified monoclonal antibodies (mAbs) capable of cross-reactive binding and neutralization of animal sarbecoviruses and SARS-CoV-2 variants by screening single mouse B cells secreting IgGs that bind two or more sarbecovirus RBDs. Single-particle cryo-EM structures of antibody-spike complexes, including a Fab-Omicron complex, mapped neutralizing mAbs to conserved class 1/4 RBD epitopes. Structural analyses revealed neutralization mechanisms, potentials for intra-spike trimer cross-linking by IgGs, and induced changes in trimer upon Fab binding. In addition, we identified a mAb-resembling Bebtelovimab, an EUA-approved human class 3 anti-RBD mAb. These results support using mosaic RBD-nanoparticle vaccination to generate and identify therapeutic pan-sarbecovirus and pan-variant mAbs.

Intermediate conformations of CD4-bound HIV-1 Env heterotrimers.

HIV-1 envelope (Env) exhibits distinct conformational changes in response to host receptor (CD4) engagement. Env, a trimer of gp120 and gp41 heterodimers, has been structurally characterized in a closed, prefusion conformation with closely associated gp120s and coreceptor binding sites on gp120 V3 hidden by V1V2 loops1-4 and in fully saturated CD4-bound open Env conformations with changes including outwardly rotated gp120s and displaced V1V2 loops3-9. To investigate changes resulting from substoichiometric CD4 binding, we solved single-particle cryo-electron microscopy (cryo-EM) structures of soluble, native-like heterotrimeric Envs bound to one or two CD4 molecules. Most of the Env trimers bound to one CD4 adopted the closed, prefusion Env state, with a minority exhibiting a heterogeneous partially open Env conformation. When bound to two CD4s, the CD4-bound gp120s exhibited an open Env conformation including a four-stranded gp120 bridging sheet and displaced gp120 V1V2 loops that expose the coreceptor sites on V3. The third gp120 adopted an intermediate, occluded-open state10 that showed gp120 outward rotation but maintained the prefusion three-stranded gp120 bridging sheet with only partial V1V2 displacement and V3 exposure. We conclude that most of the engagements with one CD4 molecule were insufficient to stimulate CD4-induced conformational changes, whereas binding two CD4 molecules led to Env opening in CD4-bound protomers only. The substoichiometric CD4-bound soluble Env heterotrimer structures resembled counterparts derived from a cryo-electron tomography study of complexes between virion-bound Envs and membrane-anchored CD4 (ref. 11), validating their physiological relevance. Together, these results illuminate intermediate conformations of HIV-1 Env and illustrate its structural plasticity.

Consolidation chemotherapy after definitive concurrent chemoradiotherapy in patients with inoperable esophageal squamous cell carcinoma: a multicenter non-inferiority phase III randomized clinical trial.

BACKGROUND: Definitive concurrent chemoradiotherapy (dCCRT) is the gold standard for the treatment of locally advanced esophageal squamous cell carcinoma (ESCC). However, the potential benefits of consolidation chemotherapy after dCCRT in patients with esophageal cancer remain debatable. Prospective randomized controlled trials comparing the outcomes of dCCRT with or without consolidation chemotherapy in patients with ESCC are lacking. In this study, we aim to generate evidence regarding consolidation chemotherapy efficacy in patients with locally advanced, inoperable ESCC. METHODS: This is a multicenter, prospective, open-label, phase-III randomized controlled trial comparing non-inferiority of dCCRT alone to consolidation chemotherapy following dCCRT. In total, 600 patients will be enrolled and randomly assigned in a 1:1 ratio to receive either consolidation chemotherapy after dCCRT (Arm A) or dCCRT alone (Arm B). Overall survival will be the primary endpoint, whereas progression-free survival, locoregional progression-free survival, distant metastasis-free survival, and treatment-related toxicity will be the secondary endpoints. DISCUSSION: This study aid in further understanding the effects of consolidation chemotherapy after dCCRT in patients with locally advanced, inoperable ESCC. TRIAL REGISTRATION: ChiCTR1800017646.

TMEM79 Ameliorates Cerebral Ischemia/Reperfusion Injury Through Regulating Inflammation and Oxidative Stress via the Nrf2/NLRP3 Pathway.

BACKGROUND: Cerebral ischemia/reperfusion injury (CIRI) is still a complicated disease with high fatality rates worldwide. Transmembrane Protein 79 (TMEM79) regulates inflammation and oxidative stress in some other diseases. METHODS: CIRI mouse model was established using C57BL/6J mice through middle cerebral artery occlusion-reperfusion (MCAO/R), and BV2 cells were subjected to oxygen and glucose deprivation/reoxygenation (OGD/R) to simulate CIRI. Brain tissue or BV2 cells were transfected or injected with lentivirus-carried TMEM79 overexpression vector. The impact of TMEM79 on CIRI-triggered oxidative stress was ascertained by dihydroethidium (DHE) staining and examination of oxidative stress indicators. Regulation of TMEM79 in neuronal apoptosis and inflammation was determined using TUNEL staining and ELISA. RESULTS: TMEM79 overexpression mitigated neurological deficit induced by MCAO/R and decreased the extent of cerebral infarct. TMEM79 prevented neuronal death in brain tissue of MCAO/R mouse model and suppressed inflammatory response by reducing inflammatory cytokines levels. Moreover, TMEM79 significantly attenuated inflammation and oxidative stress caused by OGD/R in BV2 cells. TMEM79 facilitated the activation of Nrf2 and inhibited NLRP3 and caspase-1 expressions. Rescue experiments indicated that the Nrf2/NLRP3 signaling pathway mediated the mitigative effect of TMEM79 on CIRI in vivo and in vitro. CONCLUSION: Overall, TMEM79 was confirmed to attenuate CIRI via regulating the Nrf2/NLRP3 signaling pathway.

EEG-Based Feature Classification Combining 3D-Convolutional Neural Networks with Generative Adversarial Networks for Motor Imagery.

BACKGROUND: The adoption of convolutional neural networks (CNNs) for decoding electroencephalogram (EEG)-based motor imagery (MI) in brain-computer interfaces has significantly increased recently. The effective extraction of motor imagery features is vital due to the variability among individuals and temporal states. METHODS: This study introduces a novel network architecture, 3D-convolutional neural network-generative adversarial network (3D-CNN-GAN), for decoding both within-session and cross-session motor imagery. Initially, EEG signals were extracted over various time intervals using a sliding window technique, capturing temporal, frequency, and phase features to construct a temporal-frequency-phase feature (TFPF) three-dimensional feature map. Generative adversarial networks (GANs) were then employed to synthesize artificial data, which, when combined with the original datasets, expanded the data capacity and enhanced functional connectivity. Moreover, GANs proved capable of learning and amplifying the brain connectivity patterns present in the existing data, generating more distinctive brain network features. A compact, two-layer 3D-CNN model was subsequently developed to efficiently decode these TFPF features. RESULTS: Taking into account session and individual differences in EEG data, tests were conducted on both the public GigaDB dataset and the SHU laboratory dataset. On the GigaDB dataset, our 3D-CNN and 3D-CNN-GAN models achieved two-class within-session motor imagery accuracies of 76.49% and 77.03%, respectively, demonstrating the algorithm's effectiveness and the improvement provided by data augmentation. Furthermore, on the SHU dataset, the 3D-CNN and 3D-CNN-GAN models yielded two-class within-session motor imagery accuracies of 67.64% and 71.63%, and cross-session motor imagery accuracies of 58.06% and 63.04%, respectively. CONCLUSIONS: The 3D-CNN-GAN algorithm significantly enhances the generalizability of EEG-based motor imagery brain-computer interfaces (BCIs). Additionally, this research offers valuable insights into the potential applications of motor imagery BCIs.

Solidification/stabilization and optimization of municipal solid waste incineration fly ash with aluminosilicate solid wastes.

Alkali-activation is an effective municipal solid waste incineration fly ash (MSWIFA) solidification/stabilization (S/S) technology. However, the characteristics of calcium-rich silica-poor aluminum phase in MSWIFA easily cause the structural instability and contamination of alkali activated MSWIFA S/S bodies. Therefore, the aluminosilicate solid wastes are used in this work to optimize the immobilization and structural properties. Results showed that incorporation of aluminosilicate solid wastes significantly improved the compressive strength and heavy metals pollution toxicity of MSWIFA S/S bodies. Compared to alkali activated MSWIFA, the compressive strength of S/S bodies with addition of coal fly ash, silica fume and granulated blast furnace slag improved by 31.0%, 47.6% and 50.8% when the curing time was 28 days, respectively. Leachability of Pb, Zn and Cd in these alkali activated MSWIFA S/S bodies was far below the threshold value specified in Standard GB16889. Aluminosilicate solid wastes provided abundant Si/Al structural units, and some new phases such as ettringite(AFt, 3CaO⋅Al2O3⋅3CaSO4⋅32H2O), calcium sulfoaluminate hydrate (3CaO⋅Al2O3⋅CaSO4⋅12H2O) and Friedel's salt (CaO⋅Al2O3⋅CaCl2⋅10H2O) can be detected in S/S matrix with aluminosilicate solid wastes, along comes increased the amount of the amorphous phases. Lower Ca/Si molar ratio tended to form the network structure gel similar to tobermorite with higher polymerization degree. Meanwhile, the silica tetrahedron of the gels changed from the oligomerization state like island to the hyperomerization state like chain, layer network or three-dimensional structure, and average molecular chain length increased. These findings provide theoretical basis for structural properties optimization and resource utilization of MSWIFA S/S matrices.

A structural framework for unidirectional transport by a bacterial ABC exporter.

The ATP-binding cassette (ABC) transporter of mitochondria (Atm1) mediates iron homeostasis in eukaryotes, while the prokaryotic homolog from Novosphingobium aromaticivorans (NaAtm1) can export glutathione derivatives and confer protection against heavy-metal toxicity. To establish the structural framework underlying the NaAtm1 transport mechanism, we determined eight structures by X-ray crystallography and single-particle cryo-electron microscopy in distinct conformational states, stabilized by individual disulfide crosslinks and nucleotides. As NaAtm1 progresses through the transport cycle, conformational changes in transmembrane helix 6 (TM6) alter the glutathione-binding site and the associated substrate-binding cavity. Significantly, kinking of TM6 in the post-ATP hydrolysis state stabilized by MgADPVO4 eliminates this cavity, precluding uptake of glutathione derivatives. The presence of this cavity during the transition from the inward-facing to outward-facing conformational states, and its absence in the reverse direction, thereby provide an elegant and conceptually simple mechanism for enforcing the export directionality of transport by NaAtm1. One of the disulfide crosslinked NaAtm1 variants characterized in this work retains significant glutathione transport activity, suggesting that ATP hydrolysis and substrate transport by Atm1 may involve a limited set of conformational states with minimal separation of the nucleotide-binding domains in the inward-facing conformation.

UDP-glucuronosyltransferase is involved in susceptibility of Chironomus kiiensis Tokunaga, 1936 (Diptera: Chironomidae) to insecticides.

UDP-glucuronosyltransferases (UGTs) could transform various exogenous and endogenous compounds, which help detoxification of pesticides in insects. To investigate the role of UGTs in the detoxification metabolism of insecticides in Chironomus kiiensis, CkUGT302M1, CkUGT302N1, CkUGT308N1 and CkUGT36J1 genes were identified with 1449-1599 bp encoding 482-532 amino acids. Four UGT genes shared 40.86∼53.36% identity with other homologous insect species, and expressed in all developmental stages, notably in the larval and adult stages. Expression of CkUGTs was higher in the gastric caecum, midgut and head. Moreover, CkUGTs expression and activity were significantly increased in C. kiiensis larvae in exposure to sublethal concentrations of carbaryl, deltamethrin and phoxim, respectively. To further explore the functions of UGT genes, the CkUGT308N1 was effectively silenced in 4th instar C. kiiensis larvae by RNA interference, which resulted in the mortality of dsCkUGT308N1 treated larvae increased by 71.43%, 111.11% and 62.50% under sublethal doses of carbaryl, deltamethrin and phoxim at the 24-h time point, respectively. The study revealed that the CkUGT308N1 gene in C. kiiensis could contribute to the metabolism of pesticides and provide a scientific basis for evaluating the water pollution of pesticides.

Solidification of municipal solid waste incineration fly ash with alkali-activated technology.

Alkali-activation is effective municipal solid waste incineration fly ash (MSWIFA) solidification/stabilization (S/S) technology. Percolation and migration of heavy metals in MSWIFA S/S matrix is a complicated and slow process. Here, several alkali-activated MSWIFA samples are selected to comparatively investigate the long-term leaching behavior and environmental availability of Pb, Zn and Cd when exposed in different erosion environment. Acid environment posed the more serious destroy to MSWIFA S/S matrices. RAC demonstrated that potential risk level of heavy metals is higher in acid rain environment, and Cd, Zn showed the prominent risk. When soaked in acid rain solution, the surface of alkali-activated MSWIFA S/S matrices was cracked seriously and a large number of hardened slurry peeled off. However, more stable structural properties and lower heavy metal leachability can be found in alkali-activated MSWIFA/aluminosilicate. The immobilization efficiency of Pb, Zn and Cd were all above 99.0%. Microstructure and morphology results indicated that there is new phase Friedel's salts generated and much more amorphous substance such as C-(A)-S-H gel with incorporation of aluminosilicate, which all contributed much to the formation of compact and stable microstructure, then significantly facilitated the encapsulation of heavy metal. These findings will provide theoretical basis and new insight for resource utilization and security landfill of MSWIFA.

A comparative study on characteristics and leaching toxicity of fluidized bed and grate furnace MSWI fly ash.

Grate furnace and fluidized bed are the most widely used technologies for municipal solid waste incineration (MSWI), which play significant roles in characteristics of MSWI fly ash. A comparative study of the physicochemical characteristics, microstructure morphology and leaching toxicity of fluidized bed and grate furnace MSWI fly ash was conducted in this work, and some resource utilization and disposal treatments were proposed. Results showed that calcium salt and chlorine salt formed the dominant components of MSWI fly ash. CaO-SiO2-Al2O3 ternary system indicated that MSWI fly ash had potential pozzolanic activity, similar to coal fly ash and blast furnace slag. The total Pb, Cd and Zn contents in fluidized bed MSWI fly ash was only 1/2, 1/3 and 2/3 of grate furnace MSWI fly ash, respectively. Leachability of Pb in MSWI fly ash collected from Dalian, Shanghai, Zhuji and Hangzhou was 4.25 mg/L, 3.83 mg/L, 3.84 mg/L and 3.68 mg/L, 28.3, 25.5, 25.6 and 24.5 times as much as the national standard limitation (GB16889-2008), respectively. However, grate furnace MSWI fly ash with high chloride and unstable chemical speciation distribution of heavy metals would pose more environmental risk toits immobilization and disposal. Fluidized bed fly ash is a promising candidate for preparing the Portland cement clinker, microcrystalline glass and ceramics due to its high Si and Al content. Grate furnace MSWI fly ash is more appropriate for Alinite cement preparation because of high chloride content.

Neoadjuvant chemotherapy versus neoadjuvant chemoradiotherapy for locally advanced oesophageal squamous cell carcinoma: a single-Centre, open-label, randomized, controlled, clinical trial (HCHTOG1903).

BACKGROUND: Neoadjuvant therapy plus oesophagectomy has been accepted as the standard treatment for patients with potentially curable locally advanced oesophageal cancer. No completed randomized controlled trial (RCT) has directly compared neoadjuvant chemotherapy and neoadjuvant chemoradiation in patients with oesophageal squamous cell carcinoma (ESCC). The aim of the current RCT is to investigate the impact of neoadjuvant chemotherapy plus surgery and neoadjuvant chemoradiotherapy plus surgery on overall survival for patients with resectable locally advanced ESCC. METHODS: This open label, single-centre, phase III RCT randomized patients (cT2-T4aN + M0 and cT3-4aN0M0) in a 1:1 fashion to receive either the CROSS regimen (paclitaxel 50 mg/m2; carboplatin (area under the curve = 2), q1w, 5 cycles; and concurrent radiotherapy, 41.4 Gy/23 F, over 5 weeks) or neoadjuvant chemotherapy (paclitaxel 175 mg/m2; and cisplatin 75 mg/m2, q21d, 2 cycles). Assuming a 12% 5-year overall survival difference in favour of the CROSS regimen, 80% power with a two-sided alpha level of 0.05 and a 5% dropout each year for an estimated 3 years enrolment, the power calculation requires 456 patients to be recruited (228 in each group). The primary endpoint is 5-year overall survival, with a minimum 5-year follow-up. The secondary endpoints include 5-year disease-free survival, toxicity, pathological complete response rate, postoperative complications, postoperative mortality and quality of life. A biobank of pre-treatment and resected tumour tissue will be built for translational research in the future. DISCUSSION: This RCT directly compares a neoadjuvant chemotherapy regimen with a standard CROSS regimen in terms of overall survival for patients with locally advanced ESCC. The results of this RCT will provide an answer for the controversy regarding the survival benefits between the two treatment strategies. TRIAL REGISTRATION: NCT04138212, date of registration: October 24, 2019.

On-Ground Processing of Yaogan-24 Remote Sensing Satellite Attitude Data and Verification Using Geometric Field Calibration.

Satellite attitude accuracy is an important factor affecting the geometric processing accuracy of high-resolution optical satellite imagery. To address the problem whereby the accuracy of the Yaogan-24 remote sensing satellite's on-board attitude data processing is not high enough and thus cannot meet its image geometry processing requirements, we developed an approach involving on-ground attitude data processing and digital orthophoto (DOM) and the digital elevation model (DEM) verification of a geometric calibration field. The approach focuses on three modules: on-ground processing based on bidirectional filter, overall weighted smoothing and fitting, and evaluation in the geometric calibration field. Our experimental results demonstrate that the proposed on-ground processing method is both robust and feasible, which ensures the reliability of the observation data quality, convergence and stability of the parameter estimation model. In addition, both the Euler angle and quaternion could be used to build a mathematical fitting model, while the orthogonal polynomial fitting model is more suitable for modeling the attitude parameter. Furthermore, compared to the image geometric processing results based on on-board attitude data, the image uncontrolled and relative geometric positioning result accuracy can be increased by about 50%.

Neoadjuvant treatment combined with planned surgery in laryngeal function preservation for locally advanced pyriform sinus carcinoma.

OBJECTIVES: This study aimed to determine the efficacy and feasibility of preoperative radiotherapy with or without chemotherapy (XRT) followed by surgery of locally advanced pyriform sinus carcinoma. MATERIALS AND METHODS: We performed a retrospective study of 482 patients with T3 and T4 pyriform sinus carcinoma treated with curative intent between 1979 and 2008. The patients were divided into 4 groups according to different treatment patterns. RESULTS: The 5-year disease-special survival rate was 32%. Survival was best for the patients treated with surgery followed by XRT (41.1%) and with neoadjuvant XRT followed by surgery (39.4%), but it was worse with surgery only (27.1%) and XRT alone (23%). The disease-special survival was significantly different between the patients who accepted multidisciplinary therapy and those who accepted single therapy (P<0.001). The 5-year laryngeal function preservation survival rate of the patients with XRT followed by surgery was 13.6%, which was similar to those with XRT alone (16.2%), and superior to those who accepted surgery followed by XRT (3.8%) and surgery alone (0). Multivariate analysis results revealed that treatment pattern is an independent predictor of both overall and laryngeal function preservation survival rates (P < 0.001; hazard ratio, 0.56; 95% confidence interval, 0.44-0.71). Perioperative wound complications were not different among the patients in the 3 groups who accepted surgical treatment (P > 0.05). CONCLUSIONS: The results suggested that neoadjuvant XRT followed by planned surgery was feasible with satisfactory oncological and functional outcomes.

The Fat Body-Specific GST Gene SlGSTe11 Enhances the Tolerance of Spodoptera litura to Cyantraniliprole and Nicotine.

Spodoptera litura is a significant agricultural pest, and its glutathione S-transferase (GST) plays a crucial role in insecticide resistance. This study aimed to investigate the relationship between the SlGSTe11 gene of S. litura and resistance to cyantraniliprole and nicotine. Transcriptome analysis revealed that SlGSTe11 is highly expressed mainly in fat bodies, with a significant increase in SlGSTe11 gene expression under induction by cyantraniliprole and nicotine. The ectopic expression of the SlGSTe11 gene in transgenic fruit flies resulted in a 5.22-fold increase in the tolerance to cyantraniliprole. Moreover, compared to the UAS-SlGSTe11 line, the Act5C-UAS>SlGSTe11 line laid more eggs and had a lower mortality after nicotine exposure. RNAi-mediated inhibition of SlGSTe11 gene expression led to a significant increase in the mortality of S. litura under cyantraniliprole exposure. In vitro metabolism experiments demonstrated that the recombinant SlGSTe11 protein efficiently metabolizes cyantraniliprole. Molecular docking results indicated that SlGSTe11 has a strong affinity for both cyantraniliprole and nicotine. These findings suggest that SlGSTe11 is involved in the development of resistance to cyantraniliprole and nicotine in S. litura.

Comparative analysis of codon usage bias in the chloroplast genomes of eighteen Ampelopsideae species (Vitaceae).

BACKGROUND: The tribe Ampelopsideae plants are important garden plants with both medicinal and ornamental values. The study of codon usage bias (CUB) facilitates a deeper comprehension of the molecular genetic evolution of species and their adaptive strategies. The joint analysis of CUB in chloroplast genomes (cpDNA) offers valuable insights for in-depth research on molecular genetic evolution, biological resource conservation, and elite breeding within this plant family. RESULTS: The base composition and codon usage preferences of the eighteen chloroplast genomes were highly similar, with the GC content of bases at all positions of their codons being less than 50%. This indicates that they preferred A/T bases. Their effective codon numbers were all in the range of 35-61, which indicates that the codon preferences of the chloroplast genomes of the 18 Ampelopsideae plants were relatively weak. A series of analyses indicated that the codon preference of the chloroplast genomes of the 18 Ampelopsideae plants was influenced by a combination of multiple factors, with natural selection being the primary influence. The clustering tree generated based on the relative usage of synonymous codons is consistent with some of the results obtained from the phylogenetic tree of chloroplast genomes, which indicates that the clustering tree based on the relative usage of synonymous codons can be an important supplement to the results of the sequence-based phylogenetic analysis. Eventually, 10 shared best codons were screened on the basis of the chloroplast genomes of 18 species. CONCLUSION: The codon preferences of the chloroplast genome in Ampelopsideae plants are relatively weak and are primarily influenced by natural selection. The codon composition of the chloroplast genomes of the eighteen Ampelopsideae plants and their usage preferences were sufficiently similar to demonstrate that the chloroplast genomes of Ampelopsideae plants are highly conserved. This study provides a scientific basis for the genetic evolution of chloroplast genes in Ampelopsideae species and their suitable strategies.

CF2-II Alternative Splicing Isoform Regulates the Expression of Xenobiotic Tolerance-Related Cytochrome P450 CYP6CY22 in Aphis gossypii Glover.

The expression of P450 genes is regulated by trans-regulatory factors or cis-regulatory elements and influences how endogenous or xenobiotic substances are metabolized in an organism's tissues. In this study, we showed that overexpression of the cytochrome P450 gene, CYP6CY22, led to resistance to cyantraniliprole in Aphis gossypii. The expression of CYP6CY22 increased in the midgut and remaining carcass of the CyR strain, and after repressing the expression of CYP6CY22, the mortality of cotton aphids increased 2.08-fold after exposure to cyantraniliprole. Drosophila ectopically expressing CYP6CY22 exhibited tolerance to cyantraniliprole and cross-tolerance to xanthotoxin, quercetin, 2-tridecanone, tannic acid, and nicotine. Moreover, transcription factor CF2-II (XM_027994540.2) is transcribed only as the splicing variant isoform CF2-II-AS, which was found to be 504 nucleotides shorter than CF2-II in A. gossypii. RNAi and yeast one-hybrid (Y1H) results indicated that CF2-II-AS positively regulates CYP6CY22 and binds to cis-acting element p (-851/-842) of CYP6CY22 to regulate its overexpression. The above results indicated that CYP6CY22 was regulated by the splicing isoform CF2-II-AS, which will help us further understand the mechanism of transcriptional adaption of cross-tolerance between synthetic insecticides and plant secondary metabolites mediated by P450s.

Natural Product-Inspired Discovery of Naphthoquinone-Furo-Piperidine Derivatives as Novel STAT3 Inhibitors for the Treatment of Triple-Negative Breast Cancer.

Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer, and STAT3 has emerged as an effective drug target for TNBC treatment. Herein, we employed a scaffold-hopping strategy of natural products to develop a series of naphthoquinone-furopiperidine derivatives as novel STAT3 inhibitors. The in vitro assay showed that compound 10g possessed higher antiproliferative activity than Cryptotanshinone and Napabucasin against TNBC cell lines, along with lower toxicity and potent antitumor activity in a TNBC xenograft model. Mechanistically, 10g could inhibit the phosphorylation of STAT3 and the binding affinity was determined by the SPR assay (KD = 8.30 µM). Molecule docking studies suggested a plausible binding mode between 10g and the SH2 domain, in which the piperidine fragment and the terminal hydroxy group of 10g played an important role in demonstrating the success of this evolution strategy. These findings provide a natural product-inspired novel STAT3 inhibitor for TNBC treatment.