Cyclic immonium ion of lactyllysine reveals widespread lactylation in the human proteome.

Lactylation was initially discovered on human histones. Given its nascence, its occurrence on nonhistone proteins and downstream functional consequences remain elusive. Here we report a cyclic immonium ion of lactyllysine formed during tandem mass spectrometry that enables confident protein lactylation assignment. We validated the sensitivity and specificity of this ion for lactylation through affinity-enriched lactylproteome analysis and large-scale informatic assessment of nonlactylated spectral libraries. With this diagnostic ion-based strategy, we confidently determined new lactylation, unveiling a wide landscape beyond histones from not only the enriched lactylproteome but also existing unenriched human proteome resources. Specifically, by mining the public human Meltome Atlas, we found that lactylation is common on glycolytic enzymes and conserved on ALDOA. We also discovered prevalent lactylation on DHRS7 in the draft of the human tissue proteome. We partially demonstrated the functional importance of lactylation: site-specific engineering of lactylation into ALDOA caused enzyme inhibition, suggesting a lactylation-dependent feedback loop in glycolysis.

Carbene Footprinting Directs Design of Genetically Encoded Proximity-Reactive Protein Binders.

Genetically encoding proximal-reactive unnatural amino acids (PrUaas), such as fluorosulfate-l-tyrosine (FSY), into natural proteins of interest (POI) confer the POI with the ability to covalently bind to its interacting proteins (IPs). The PrUaa-incorporated POIs hold promise for blocking undesirable POI-IP interactions. Selecting appropriate PrUaa anchor sites is crucial, but it remains challenging with the current methodology, which heavily relies on crystallography to identify the proximal residues between the POIs and the IPs for the PrUaa anchorage. To address the challenge, here, we propose a footprinting-directed genetically encoded covalent binder (footprinting-GECB) approach. This approach employs carbene footprinting, a structural mass spectrometry (MS) technique that quantifies the extent of labeling of the POI following the addition of its IP, and thus identifies the responsive residues. By genetically encoding PrUaa into these responsive sites, POI variants with covalent bonding ability to its IP can be produced without the need for crystallography. Using the POI-IP model, KRAS/RAF1, we showed that engineering FSY at the footprint-assigned KRAS residue resulted in a KRAS variant that can bind irreversibly to RAF1. Additionally, we inserted FSY at the responsive residue in RAF1 upon footprinting the oncogenic KRASG12D/RAF1, which lacks crystal structure, and generated a covalent binder to KRASG12D. Together, we demonstrated that by adopting carbene footprinting to direct PrUaa anchorage, we can greatly expand the opportunities for designing covalent protein binders for PPIs without relying on crystallography. This holds promise for creating effective PPI inhibitors and supports both fundamental research and biotherapeutics development.

Dexmedetomidine administration during brain tumour resection for prevention of postoperative delirium: a randomised trial.

BACKGROUND: Delirium is common, especially after neurosurgery. Dexmedetomidine might reduce delirium by improving postoperative analgesia and sleep quality. We tested the primary hypothesis that dexmedetomidine administration during intracerebral tumour resection reduces the incidence of postoperative delirium. METHODS: This randomised, double-blind, placebo-controlled trial was conducted in two tertiary-care hospitals in Beijing. We randomised 260 qualifying patients to either dexmedetomidine (n=130) or placebo (n=130). Subjects assigned to dexmedetomidine were given a loading dose of 0.6 µg kg-1 followed by continuous infusion at 0.4 µg kg-1 h-1 until dural closure; subjects in the placebo group were given comparable volumes of normal saline. The primary outcome was the incidence of delirium, which was assessed with the Confusion Assessment Method twice daily during the initial 5 postoperative days. RESULTS: The average (standard deviation) age of participating patients was 45 (12) yr, duration of surgery was 4.2 (1.5) h, and patients assigned to dexmedetomidine were given an average of 126 (45) µg of dexmedetomidine. There was less delirium during the initial 5 postoperative days in patients assigned to dexmedetomidine (22%, 28 of 130 patients) than in those given placebo (46%, 60 of 130 patients) with a risk ratio of 0.51 (95% confidence interval: 0.36-0.74, P<0.001). Postoperative pain scores with movement, and recovery and sleep quality were improved by dexmedetomidine (P<0.001). The incidence of safety outcomes was similar in each group. CONCLUSIONS: Prophylactic intraoperative dexmedetomidine infusion reduced by half the incidence of delirium during the initial 5 postoperative days in patients recovering from elective brain tumour resection. CLINICAL TRIAL REGISTRATION: NCT04674241.

Living Cell-Target Responsive Accessibility Profiling Reveals Silibinin Targeting ACSL4 for Combating Ferroptosis.

We report a living cell-target responsive accessibility profiling (LC-TRAP) approach to identify the targetome of silibinin (SIL), a well-established hepatoprotective natural product (NP), in HepG2 cells. Proteins showing accessibility changes, probed by covalent lysine labeling reagents and leveraged by multiplexed quantitative proteomics, following the administration of SIL to the living cells were assigned as potential targets. Among the assigned targetome, ACSL4, an enzyme essential for ferroptosis induction, might be involved in the hepatoprotective effects of SIL and hence was intensively validated. We first demonstrated that SIL protected HepG2 cells from ferroptosis dependent on ACSL4. Then, we used biophysical assays and a SIL-derivatized chemical probe to corroborate that SIL can bind to ACSL4. The ensuing enzymatic assays showed that SIL inhibited ACSL4 enzymatic activity, thereby mitigating the ACSL4-mediated ferroptosis. As such, we revealed that ACSL4 inhibition, using SIL as a model compound, represents a promising hepatoprotective strategy. Further, since TRAP probes the accessibility changes of reactive proteinaceous lysines, it can pinpoint the proximal regions where the ligand engagement may occur. Thus, the LC-TRAP analysis of SIL, the newly discovered ligand of ACSL4, and arachidonic acid (AA), the substrate, intriguingly showed that SIL and AA both affected the conformation of the K536-proximal region of ACSL4, albeit through distinct binding patterns. Collectively, we describe a straightforward LC-TRAP workflow that does not involve ligand-derived probe synthesis and is widely applicable to target discovery of NPs.

The role of clinical characteristics and pulmonary function testing in predicting risk of pneumothorax by CT-guided percutaneous core needle biopsy of the lung.

BACKGROUND: We aim to analyze the risk factors for pneumothorax associated with computed tomography (CT)-guided percutaneous core needle biopsy (PCNB) of the lung. Whether the lung function characteristics are related to pneumothorax is unclear. METHODS: We retrospectively evaluated 343 patients who received CT-guided pulmonary PCNBs and underwent preoperative pulmonary function testing. Demographical, lesion-related, procedure-related features and histopathological diagnosis, as well as results of pulmonary function test were analyzed as risk factors of pneumothorax RESULTS: Variables associated with higher rate of pneumothorax were location of lesion, presence of emphysema, and dwell time. The proportion of middle lobe, lingular, or lower lobe lesions in pneumothorax group (30/50, 60.0%) is higher than non-pneumothorax group (113/293, 38.6%). The incidence of emphysema in pneumothorax group was significantly higher than that in non-pneumothorax group (34.0% vs. 7.5%). Obstructive pulmonary function abnormalities, not restrictive, mixed ventilation function abnormalities and small airway dysfunction, correlated with pneumothorax. Multivariate logistic regression analysis showed lower location of lesion sampled and presence of emphysema were independent predictors of pneumothorax. Although dwell time, FEV1/FVC ratio, FEF50%, FEF75% and FEF25-75% were significantly correlated with pneumothorax on univariate analysis, these were not confirmed to be independent predictors. CONCLUSIONS: Patients with obstructive pulmonary dysfunction have a higher risk of pneumothorax. Presence of emphysema was the most important predictor of pneumothorax, followed by location of lesion.

Rapid Hemostatic Biomaterial from a Natural Bath Sponge Skeleton.

Uncontrolled bleeding is the main cause of mortality from trauma. Collagen has been developed as an important hemostatic material due to its platelet affinity function. A bath sponge skeleton is rich in collagen, also known as spongin. To understand the hemostatic effect of spongin, spongin materials, SX, SFM and SR were prepared from the bath sponge Spongia officinalis, and hemostatic experiments were performed. The SX, SFM and SR were significantly better than the positive control, type I collagen, in shortening the whole blood clotting time in vitro and hemostasis upon rat tail amputation. In a hemostatic experiment of rabbit common carotid artery injury, the hemostatic time and 3 h survival rate of the SFM group were 3.00 ± 1.53 min and 100%, respectively, which are significantly better than those of the commercial hemostat CELOX-A (10.33 ± 1.37 min and 67%, respectively). Additionally, the SFM showed good coagulation effects in platelet-deficient blood and defibrinated blood, while also showing good biocompatibility. Through a variety of tests, we speculated that the hemostatic activity of the SFM is mainly caused by its hyperabsorbency, high affinity to platelets and high effective concentration. Overall, the SFM and spongin derivates could be potential hemostatic agents for uncontrolled bleeding and hemorrhagic diseases caused by deficiency or dysfunction of coagulation factors.

Genetic diversity within grouper species and a method for interspecific hybrid identification using DNA barcoding and RYR3 marker.

Groupers (family Epinephelidae) are an assemblage of coral reef fishes comprising more than 160 species in 16 genera, many of which are both environmentally and economically valuable. Because of their similar morphology, variable color patterns, and tendency for interspecies hybridization, morphological identification of groupers usually leads to taxonomic confusion. To find an effective method for identifying different grouper species and hybrids, evaluate genetic diversity and uncover any synonymous or cryptic species, we sampled a total of 221 specimens representing 57 species in 9 genera in the China Seas. Both mitochondrial (mt) cytochrome oxidase subunit I (COI) and NADH dehydrogenase subunit 2 (ND2) were found to be effective barcoding genes. We also developed an efficient protocol for identifying hybrid groupers using mt markers and the nuclear RYR3 gene and found the first record of wide interspecies hybridization in genus Epinephelus. This barcoding study revealed high genetic divergence in many widespread species and possible synonyms. In addition to providing a molecular method for identifying grouper species, this study offers important resources for the further study of grouper conservation genetics, speciation, hybridization and other evolutionary traits.

Skin Delivery of Hydrophilic Biomacromolecules Using Marine Sponge Spicules.

We report the development of sponge Haliclona sp. spicules, referred to as SHS, and its topical application in skin delivery of hydrophilic biomacromolecules, a series of fluorescein isothiocyanate-dextrans (FDs). SHS are silicious oxeas which are sharp-edged and rod-shaped (∼120 µm in length and ∼7 µm in diameter). SHS can physically disrupt skin in a dose-dependent manner and retain within the skin over at least 72 h, which allows sustained skin penetration of hydrophilic biomacromolecules. The magnitude of enhancement of FD delivery into skin induced by SHS treatment was dependent on its molecular weight. Specifically, SHS topical application enhanced FD-10 (MW: 10 kDa) penetration into porcine skin in vitro by 33.09 ± 7.16-fold compared to control group (p < 0.01). SHS dramatically increased the accumulation of FD-10 into and across the dermis by 62.32 ± 13.48-fold compared to the control group (p < 0.01). In vivo experiments performed using BALB/c mice also confirmed the effectiveness of SHS topical application; the skin absorption of FD-10 with SHS topical application was 72.14 ± 48.75-fold (p < 0.05) and 15.39 ± 9.91-fold (p < 0.05) higher than those from the PBS and Dermaroller microneedling, respectively. Further, skin irritation study and transepidermal water loss (TEWL) measurement using guinea pig skin in vivo indicated that skin disruption induced by SHS treatment is self-limited and can be recovered with time and efficiently. SHS can offer a safe, effective, and sustained skin delivery of hydrophilic biomacromolecules and presents a promising platform technology for a wide range of cosmetic and medical applications.

Isolation and diversity of natural product biosynthetic genes of cultivable bacteria associated with marine sponge Mycale sp. from the coast of Fujian, China.

The marine sponge Mycale sp., a potential source of natural bioactive products, is widely distributed along the coast of Fujian, China. The cultivable bacterial community associated with Mycale sp., the antibacterial activities, and the PKS (polyketide synthase) and NRPS (nonribosomal peptide synthetase) gene diversity of these bacteria were investigated. Phylogenetic analysis of the 16S rRNA gene showed that the 51 isolates from Mycale sp. belonged to Actinobacteria, Bacteroidetes, Gammaproteobacteria, Alphaproteobacteria, and Firmicutes. Among them, some bacteria were first isolated from marine sponge. The 20 isolates with antimicrobial activities were primarily clustered within the groups Actinobacteria, Gammaproteobacteria, and Bacillus. Strain HNS054, which showed 99% similarity to Streptomyces labedae, exhibited the strongest antimicrobial activity against Gram-positive bacteria (Staphylococcus aureus MTCC 1430, Bacillus subtilis MTCC 441) and Vibrio species. The screening of natural product biosynthetic genes revealed that 8 Actinobacteria species with antimicrobial activities possessed PKS-KS (ketosynthase) or NRPS-A domains, and the Nocardiopsis species contained a hybrid or mixed PKS-NRPS system. The phylogenetic analysis of the amino acid sequences indicated that the identified KS domains clustered with those from diverse bacterial groups, including Actinobacteria, Alphaproteobacteria, Cyanobacteria, and Firmicutes. Most KS domain sequences had high homology (>80%) to type I KSs, but the KS domain of Nocardiopsis sp. strain HNS048 had 77% similarity to the type II KS domain of Burkholderia gladioli. The NRPS-A domains of the 8 isolates were grouped into the Gammaproteobacteria, Actinobacteria, and Firmicutes groups. The NRPS-A gene of strain HNS052, identified as Nocardiopsis cyriacigeorgica, showed only 54% similarity to Rhodococcus opacus. All results suggested that Mycale sp. harboured diverse bacteria that could contribute to the production of novel bioactive substances in the future.

Bioaccumulation of heavy metals in plant leaves from Yan׳an city of the Loess Plateau, China.

Urban plants are capable of reducing environmental pollutions through bioaccumulation contaminants in their tissues. The accumulation of heavy metals (Pb, Cu, Cd, Cr, and Zn) in leaves of nine tree species and five shrub species from Yan׳an city of China were investigated, and total metal accumulation capacities of different plants were evaluated using the metal accumulation index (MAI). The results indicated that plants in polluted environments are enriched in heavy metals relative to those in pristine environments, this is mainly caused by traffic emissions and coal combustion. Species with the highest accumulation of a single metal did not have the highest total metal accumulation capacity, the MAI should be an important indicator for tree species selection in phytoextraction and urban greening. Considering total accumulation capacities, Sabina chinensis, Juniperus formosana, Ailanthus altissima and Salix matsudana var. matsudana could be widely used in the Loess Plateau.

Effects of thinning intensities on soil infiltration and water storage capacity in a Chinese pine-oak mixed forest.

Thinning is a crucial practice in the forest ecosystem management. The soil infiltration rate and water storage capacity of pine-oak mixed forest under three different thinning intensity treatments (15%, 30%, and 60%) were studied in Qinling Mountains of China. The thinning operations had a significant influence on soil infiltration rate and water storage capacity. The soil infiltration rate and water storage capacity in different thinning treatments followed the order of control (nonthinning): <60%, <15%, and <30%. It demonstrated that thinning operation with 30% intensity can substantially improve soil infiltration rate and water storage capacity of pine-oak mixed forest in Qinling Mountains. The soil initial infiltration rate, stable infiltration rate, and average infiltration rate in thinning 30% treatment were significantly increased by 21.1%, 104.6%, and 60.9%, compared with the control. The soil maximal water storage capacity and noncapillary water storage capacity in thinning 30% treatment were significantly improved by 20.1% and 34.3% in contrast to the control. The soil infiltration rate and water storage capacity were significantly higher in the surface layer (0~20 cm) than in the deep layers (20~40 cm and 40~60 cm). We found that the soil property was closely related to soil infiltration rate and water storage capacity.

Nutrient limitation mediates soil microbial community structure and stability in forest restoration.

Soil microorganisms are often limited by nutrients, representing an important control of heterotrophic metabolic processes. However, how nutrient limitations relate to microbial community structure and stability remains unclear, which creates a knowledge gap to understanding microbial biogeography and community changes during forest restoration. Here, we combined an eco-enzymatic stoichiometry model and high-throughput DNA sequencing to assess the potential roles of nutrient limitation on microbial community structure, assembly, and stability along a forest restoration sequence in the Qinling Mountains, China. Results showed that nutrient limitations tended to decrease during the oak forest restoration. Carbon and phosphorus limitations enhanced community dissimilarity and significantly increased bacterial alpha diversity, but not fungal diversity. Stochastic assembly processes primarily structured both bacterial (average contribution of 74.73 % and 74.17 % in bulk and rhizosheath soils, respectively) and fungal (average contribution of 77.23 % and 72.04 % in bulk and rhizosheath soils, respectively) communities during forest restoration, with nutrient limitation also contributing to the importance of stochastic processes in the bacterial communities. The migration rate (m) for bacteria was 0.19 and 0.23, respectively in both bulk soil and rhizosheath soil, and was greater than that for the fungi (m was 1.19 and 1.41, respectively), indicating a stronger dispersal limitation for fungal communities. Finally, nutrient limitations significantly affected bacterial and fungal co-occurrence with more interconnections occurring among weakly nutrient-limited microbial taxa and nutrient limitations reducing community stability when nutrient availability changed during forest restoration. Our findings highlight the fundamental effects of nutrient limitations on microbial communities and their self-regulation under changing environmental resources.

[Plant Diversity Changes and Its Driving Factors of Abandoned Land at Different Restoration Stages in the Middle of the Qinling Mountains].

The process of vegetation restoration is often accompanied by significant changes in aboveground plant diversity. To explore the driving mechanism of litter nutrient-soil nutrient-enzyme activity stoichiometry on aboveground vegetation change is of great importance for maintaining regional biodiversity conservation and ecological stability. Taking typical abandoned farmland of different restoration years （1， 8， 16， 31， and 50 a） in the Qinling Mountains as the research object， the variation characteristics of plant community diversity during vegetation restoration were analyzed through field investigation. Litter nutrients， soil nutrients， and the activities of five extracellular enzymes， including ß-1，4-glucosidase （BG）， cellobiohydrolase （CBH）， ß-1，4-N-acetylglucosaminidase （NAG）， leucine aminopeptidase （LAP）， and acid phosphatase （AP）， were determined. The characteristics of litter nutrients， soil nutrients， and enzyme stoichiometric ratios during vegetation restoration and the driving mechanism of plant diversity changes were discussed. The results showed that the plant community diversity index firstly decreased and then increased with the increase in vegetation restoration years， and the minimum was reached at 16 years after restoration. The results of principal component analysis showed that there were significant differences between total plant community diversity index and litter-soil-enzyme stoichiometric characteristics in different years of vegetation restoration. The plant community diversity index had a strong positive correlation with litter C∶P ratio and litter N∶P ratio but had a negative correlation with soil enzyme C∶P ratio （EEA C∶P）. The results of redundancy analysis showed that soil EEA C∶P had the highest explanation rate of plant diversity changes during vegetation restoration （25.93%）， followed by soil TP （5.94%）， which was the key factor regulating plant diversity changes. In conclusion， plant species and quantity increased significantly in abandoned farmland in the middle part of the Qinling Mountains at the late stage of vegetation restoration. Changes in the soil environment affected microbial metabolic activities and thus changed enzyme activities. Litter-soil-soil extracellular enzymes affected the community environment and plant diversity through feedback and regulation. EEA C∶P and TP were the main driving factors of aboveground plant diversity change during vegetation restoration.

Genetic changes in muscle protein following hybridization between Haliotis diversicolor reeve Japan and Taiwan populations revealed using a proteomic approach.

Protein expression patterns were compared in a Japan and Taiwan population of Haliotis diversicolor and in a hybrid between them using 2DE and MALDI-TOF-TOF analyses. Using the software PDQuest, 924 ± 7 protein spots were detected in the Japan population (RR), 861 ± 11 in the Taiwan population (TT), and 882 ± 9 in the F1 hybrid (TR). RR and TR were clustered together, but the distance between RR and TT was the maximum using hierarchical cluster analysis. A total of 46 gel spots were identified and a total of 15 spots matched with abalone proteins (a 33.6% identification rate). Hybrid exhibiting additivity or overdominance accounted for 73.9% of these 46 identified proteins. The 46 differentially expressed proteins were shown to be involved in major biological processes, including muscle contraction and regulation, energy metabolism, and stress response. The proteins involved in energy metabolism included adenosine triphosphate (ATP) synthase ß subunit, fructose 1, 6-bisphosphate aldolase, triosephosphate isomerase, enolase, arginine kinase, and tauropine dehydrogenase. These proteins exhibited additivity in their offspring. The proteins involved in stress responses included HSP Hsp70 (exhibiting overdominance in the offspring) and Cu/Zn-superoxide dismutase (exhibiting additivity). These results suggested that proteomic approach is suitable for analysis of heterosis and functional prediction of abalone hybridization.

Baud-rate flexible clock recovery and channel identification in OTDM realized by pulse position modulation.

We propose a novel scheme of OTDM utilizing pulse position modulation, where optical null headers (ONH) are inserted between the signal pulses periodically to allow channel identification. The ONH also achieves in-band clock distribution through the generation of high contrast pilot tone on the signal power spectra, enabling baud-rate flexible clock recovery. Using the novel scheme, clock recovery with a timing jitter of less than 200 fs is achieved at different baud rates up to 344 Gbaud. We demonstrate stable clock recovery with channel identification in 344-Gb/s OTDM transmissions over dispersion managed 3-km SMF.

Cross-sectional epidemiological survey of asthma in Jinan, China.

BACKGROUND AND OBJECTIVE: The prevalence and control of asthma in all ages are seldom known in Jinan, China. This study aims to determine the prevalence, related factors and control of asthma in populations of all ages in Jinan, China. METHODS: A cross-sectional epidemiological questionnaire survey was performed with all members from approximately 4500 randomly selected families from 60 communities in the urban area and in 40 villages in the rural area. The completed questionnaires were focused on asthma, asthma-like respiratory symptoms and asthma control. RESULTS: A total of 13,645 individuals were interviewed in the survey, and the number of effective questionnaires reached 13,419. The asthma prevalence of all ages in Jinan was 1.1%. Atopy, asthma family history, household size and age were independent factors related to asthma. The control of asthma in Jinan was suboptimal in a high proportion of patients and even worse in the rural area. CONCLUSIONS: The asthma prevalence for all ages in Jinan was relatively low compared with that in western countries; however, local asthma control fell markedly short from the goals of the Global Initiative for Asthma (GINA) criteria for asthma management, which may be attributed to the poor adherence to GINA guidelines, especially the underuse of preventative medication of inhaled corticosteroids.

Pulmonary Function Test and Obstructive Sleep Apnea Hypopnea Syndrome in Obese Adults: A Retrospective Study.

Objective: We explore risk factors related to severe obstructive sleep apnea (OSA) in obese patients, including pulmonary ventilation function, diffusion function, and impulse oscillometry (IOS) data. Methods: The medical records of 207 obese patients who were prepared to undergo bariatric surgery in a hospital from May 2020 to September 2021 were retrospectively reviewed. Polysomnography (PSG), pulmonary ventilation function, diffusion function, and IOS parameters were collected according to the ethical standards of the institutional research committee (registration number: KYLL-202008-144). Logistic regression analysis was used to analyze the related independent risk factors. Results: There were significantly statistical difference in a number of pulmonary ventilation and diffusion function parameters among the non-OSAHS group, the mild-to-moderate OSA group, and the severe OSA group. However, only airway resistance parameters R5%, R10%, R15%, R20%, R25%, and R35% increased with increasing OSA severity and were positively correlated with apnea hypopnea index (AHI). Age (P = 0.012, 1.104 (1.022, 1.192)), body mass index (P< 0.0001, 1.12 (1.057, 1.187)), gender (P = 0.003, 4.129 (1.625, 10.49)), and R25% (P = 0.007, 1.018 (1.005, 1.031)) were independent risk factors for severe OSA. In patients aged 35 to 60, RV/TLC (P = 0.029, 1.272 (1.025, 1.577)) is an independent risk factor for severe OSA. Conclusion: R25% was an independent risk factor for severe OSA in obese individuals, while RV/TLC was also an independent risk factor in those aged 35 to 60. Pulmonary function tests (PFTs), particularly IOS levels, are recommended to assess severe OSA in obese patients.

Anwuligan inhibits the progression of non-small cell lung cancer via let-7c-3p/PI3K/AKT/mTOR axis.

BACKGROUND: Anwuligan (ANW) isolated from nutmeg, also known as myristyl lignan, has attracted attention due to its therapeutic potential in diseases. However, its effect on lung cancer is still unclear. METHODS: In this study, the cytotoxicity of ANW on non-small cell lung cancer (NSCLC) cells was detected using a Cell Counting Kit-8 (CCK-8) assay. In vitro, clone formation, wound healing, and Transwell assays were used to investigate the migratory and invasive abilities of NSCLC cells after ANW treatment. The expression levels of the associated proteins were detected using Western blotting. A luciferase assay was used to validate the binding of let-7c-3p to the 3'-untranslated region (UTR) of PIK3CA. In vivo, an A549 cell xenograft mouse model was used to verify the effect of ANW on tumor growth. RESULTS: The results showed that ANW treatment (20 or 50 µM) had obvious cytotoxicity against A549 and H460 cells, suppressing cell proliferation and migration in vitro. In vivo, the growth of the implanted tumor was inhibited by ANW in a nude mouse model. The growth of NSCLC cells was also inhibited by let-7c-3p overexpression in vitro and in vivo. The inhibitory effect of ANW on the proliferation and metastasis of NSCLC cells was weakened by the downregulation of let-7c-3p, whereas it was enhanced by the overexpression of let-7c-3p; PIK3CA was the main target of let-7c-3p. CONCLUSIONS: In summary, ANW inhibits the growth and metastasis of NSCLC cells in vivo and in vitro by upregulating the expression of let-7c-3p, which can regulate the PI3K/AKT/mTOR signaling pathway. PIK3CA is the main target of let-7c-3p.

Electrochemical determination of ethylvanillin based on LaV@GAC nanocomposite.

Ethyl vanillin (EVA) is widely used as a flavor additive in foods, and sensitive monitoring of EVA is of great significance for food safety. In this paper, the biomass of gum arabic derived carbon (GAC) coated with lanthanum vanadate (LaV) was constructed for the EVA sensor based on the synergistic effects of the electrochemical catalytic ability of LaV, the enhanced electrical conductivity with the GAC coating and the oxygen-containing functional groups in LaV@GAC. The as-developed LaV@GAC sensor showed a remarkable linear range from 0.06 µM to 100 µM and a low detection limit (LOD) of 6.28 nM. The electrochemical oxidation of EVA is limited by a diffusion-controlled process involving 2 electrons and 2 protons. Moreover, the LaV@GAC sensor has good recoveries (94.5-103.05%) for the detection of EVA in real milk powder samples. The proposed LaV@GAC sensor has good repeatability, high stability, and great potential for sensitive detection of flavor additives in food.

Oxidation-Induced Protein Cross-Linking in Mammalian Cells.

A proximity-enabled protein cross-linking strategy with additional spatiotemporal control is highly desirable. Here, we report an oxidation-induced protein cross-linking strategy involving the incorporation of a vinyl thioether group into proteins in both Escherichia coli and mammalian cells via genetic code expansion. We demonstrated that vinyl thioether can be selectively induced by exogenously added oxidant or by reactive oxygen species from the cellular environment, as well as by photocatalysts, and converted into a Michael acceptor, enabling fluorescence labeling and protein cross-linking.