Preparation and Formation Mechanism Study of Antibiofilm Coating Based on Phase Transition of Glutenin.

The surface of food processing equipment is easily affected by biofilm-forming bacteria, leading to cross-contamination and food safety hazards. The critical issue is how to endow the surface of contact materials with antibacterial and antibiofilm abilities. A sustainable, stable, and antibiofilm coating was prepared by phase transition of glutenin. The disulfide bonds in glutenin were reduced by tris(2-carboxyethyl)phosphine, triggering the phase transition of glutenin. Hydrophobic interactions and intermolecular disulfide bonds may be the primary forces. Furthermore, the phase-transited products formed a nanoscale coating on the surface of stainless steel and glass under their own adhesion force and gravity. The coating exhibited good stability in harsh environments. More importantly, after 3 h of direct contact, the colony of Escherichia coli and Staphylococcus aureus decreased by one logarithm. The amount of biofilm was observed to be significantly decreased through optical microscopy and scanning electron microscopy. This article provides a foundational module for developing novel coatings.

Detection and analysis of microplastics in tissues and blood of human cervical cancer patients.

Microplastics (MPs) can enter the reproductive system and can be potentially harmful to human reproductive health. In this study, 13 types of microplastics (MPs) were identified in patient blood, cancer samples, and paracarcinoma samples using Raman spectroscopy, with polyethylene, polypropylene and polyethylene-co-polypropylene being the most abundant polymer types. Futher, cotton was also found in our study. The diversity and abundance of MPs were higher in blood samples than in cancerous tissues, and there was a significant positive correlation between diversity (p < 0.05). Furthermore, the diversity and abundance of MPs in cancerous tissues were higher than in paracancerous tissues. The dimensional sizes of MPs in these samples were also very similar, with the majority of detected MPs being smaller in size. Correlation analysis showed that patient's age correlated with the abundance of MPs in blood samples, body mass index (BMI) correlated with the abundance of MPs in cancerous tissues. Notably, the frequency with which patients consume bottled water and beverages may also increase the abundance of MPs. This study identifies for the first time the presence of MPs and cotton in cancerous and paracancerous tissues of human cervical cancer patients. This provides new ideas and basic data to study the risk relationship between MP exposure and human health.

Molecular and proteomic response of Pseudomonas fluorescens biofilm cultured on lettuce (Lactuca sativa L.) after ultrasound treatment at different intensity levels.

Ultrasonic treatment is widely used for surface cleaning of vegetables in the processing of agricultural products. In the present study, the molecular and proteomic response of Pseudomonas fluorescens biofilm cultured on lettuce was investigated after ultrasound treatment at different intensity levels. The results show that the biofilm was efficiently removed after ultrasound treatment with intensity higher than 21.06 W/cm2. However, at an intensity of less than 18.42 W/cm2, P. fluorescens was stimulated by ultrasound leading to promoted bacterial growth, extracellular protease activity, extracellular polysaccharide secretion (EPS), and synthesis of acyl-homoserine lactones (AHLs) as quorum-sensing signaling molecules. The expression of biofilm-related genes, stress response, and dual quorum sensing system was upregulated during post-treatment ultrasound. Proteomic analysis showed that ultrasound activated proteins in the flagellar system, which led to changes in bacterial tendency; meanwhile, a large number of proteins in the dual-component system began to be regulated. ABC transporters accelerated the membrane transport of substances inside and outside the cell membrane and equalized the permeability conditions of the cell membrane. In addition, the expression of proteins related to DNA repair was upregulated, suggesting that bacteria repair damaged DNA after ultrasound exposure.

Deciphering the interactions between altertoxins and glutenin based on molecular dynamic simulation: inspiration from detection.

BACKGROUND: Mycotoxin contamination of food has been gaining increasing attention. Hidden mycotoxins that interact with biological macromolecules in food could make the detection of mycotoxins less accurate, potentially leading to the underestimation of the total exposure risk. Interactions of the mycotoxins alternariol (AOH) and alternariol monomethyl ether (AME) with high-molecular glutenin were explored in this study. RESULTS: The recovery rates of AOH and AME (1, 2, and 10 µg kg-1) in three types of grains (rice, corn, and wheat) were relatively low. Molecular dynamics (MD) simulations indicated that AOH and AME bound to glutenin spontaneously. Hydrogen bonds and π-π stacking were the primary interaction forces at the binding sites. Alternariol with one additional hydroxyl group exhibited stronger binding affinity to glutenin than AME when analyzing average local ionization energy. The average interaction energy between AOH and glutenin was -80.68 KJ mol-1, whereas that of AME was -67.11 KJ mol-1. CONCLUSION: This study revealed the mechanisms of the interactions between AOH (or AME) and high-molecular glutenin using MD and molecular docking. This could be useful in the development of effective methods to detect pollution levels. These results could also play an important role in the evaluation of the toxicological properties of bound altertoxins. © 2024 Society of Chemical Industry.

Microplastics supply contaminants in food chain: non-negligible threat to health safety.

The occurrence of microplastics (MPs) and organic pollutants (OPs) residues is commonly observed in diverse environmental settings, where their interactions can potentially alter the behavior, availability, and toxicity of OPs, thereby posing risks to ecosystems. Herein, we particularly emphasize the potential for bioaccumulation and the biomagnification effect of MPs in the presence of OPs within the food chain. Despite the ongoing influx of novel information, there exists a dearth of data concerning the destiny and consequences of MPs in the context of food pollution. Further endeavors are imperative to unravel the destiny and repercussions of MPs/OPs within food ecosystems and processing procedures, aiming to gain a deeper understanding of the joint effect on human health and food quality. Nevertheless, the adsorption and desorption behavior of coexisting pollutants can be significantly influenced by MPs forming biofilms within real-world environments, including temperature, pH, and food constituents. A considerable portion of MPs tend to accumulate in the epidermis of vegetables and fruits, thus necessitating further research to comprehend the potential ramifications of MPs on the infiltration behavior of OPs on agricultural product surfaces.

Predictive value of inflammation and nutritional index in immunotherapy for stage IV non-small cell lung cancer and model construction.

Identifying individuals poised to gain from immune checkpoint inhibitor (ICI) therapies is a pivotal element in the realm of tailored healthcare. The expression level of Programmed Death Ligand 1 (PD-L1) has been linked to the response to ICI therapy, but its assessment typically requires substantial tumor tissue, which can be challenging to obtain. In contrast, blood samples are more feasible for clinical application. A number of promising peripheral biomarkers have been proposed to overcome this hurdle. This research aims to evaluate the prognostic utility of the albumin-to-lactate dehydrogenase ratio (LAR), the Pan-immune-inflammation Value (PIV), and the Prognostic Nutritional Index (PNI) in predicting the response to ICI therapy in individuals with advanced non-small cell lung cancer (NSCLC). Furthermore, the study seeks to construct a predictive nomogram that includes these markers to facilitate the selection of patients with a higher likelihood of benefiting from ICI therapy. A research initiative scrutinized the treatment records of 157 advanced NSCLC patients who received ICI therapy across two Jiangxi medical centers. The cohort from Jiangxi Provincial People's Hospital (comprising 108 patients) was utilized for the training dataset, while the contingent from Jiangxi Cancer Hospital (49 patients) served for validation purposes. Stratification was based on established LAR, PIV, and PNI benchmarks to explore associations with DCR and ORR metrics. Factorial influences on ICI treatment success were discerned through univariate and multivariate Cox regression analysis. Subsequently, a Nomogram was devised to forecast outcomes, its precision gauged by ROC and calibration curves, DCA analysis, and cross-institutional validation. In the training group, the optimal threshold values for LAR, PIV, and PNI were identified as 5.205, 297.49, and 44.6, respectively. Based on these thresholds, LAR, PIV, and PNI were categorized into high (≥ Cut-off) and low (< Cut-off) groups. Patients with low LAR (L-LAR), low PIV (L-PIV), and high PNI (H-PNI) exhibited a higher disease control rate (DCR) (P < 0.05) and longer median progression-free survival (PFS) (P < 0.05). Cox multivariate analysis indicated that PS, malignant pleural effusion, liver metastasis, high PIV (H-PIV), and low PNI (L-PNI) were risk factors adversely affecting the efficacy of immunotherapy (P < 0.05). The Nomogram model predicted a concordance index (C-index) of 0.78 (95% CI: 0.73-0.84). The areas under the ROC curve (AUC) for the training group at 6, 9, and 12 months were 0.900, 0.869, and 0.866, respectively, while the AUCs for the external validation group at the same time points were 0.800, 0.886, and 0.801, respectively. Throughout immunotherapy, PIV and PNI could act as prospective indicators for forecasting treatment success in NSCLC patients, while the devised Nomogram model exhibits strong predictive performance for patient prognoses.

Microscopic Raman-based rapid detection of submicron/nano polypropylene plastics in tea and tea beverages.

Polypropylene (PP) is suitable for a broad range of applications and represents the most extensively utilized plastic in food packaging. Micro- and nano-PP plastics are prevalent categories of microplastics (MPs). However, the majority of MPs particles currently utilized in laboratory studies are man-made polystyrene (PS) spheres, and there has been limited research on micrometer- and nanoscale PP plastic particles. This study aims to employ a top-down approach in crafting micro/nanoparticle (M/NPs) models of PP particles, ensuring their enhanced relevance to real-world environments. Micro/nano PP particles, featuring a negatively charged particle size ranging from 203 to 2101 nm, were synthesized through variations in solution concentration and volume. Simultaneously, the devised MPs model was employed to develop a Raman-based qualitative and quantitative detection method for micro/nano PP particles, considering diverse sizes and concentrations. This method integrates Raman spectroscopy and microscopy to measure PP particles with varying sizes, utilizing the coffee ring effect. The Limit of detection (LOD) for 203 nm PP reached 31.25 µg/mL, while those for 382-2101 nm PP were approximately 3.9 µg/mL. The method underwent quantitative analysis by introducing 203 nm PP nanospheres into real food media (i.e., tea beverages, tea leaves), revealing a minimum LOD of approximately 31.25 µg/mL.

Identification and Visualization of Polystyrene Microplastics/Nanoplastics in Flavored Yogurt by Raman Imaging.

The contamination of food by microplastics has garnered widespread attention, particularly concerning the health risks associated with small-sized microplastics. However, detecting these smaller microplastics in food poses challenges attributed to the complexity of food matrices and instrumental and method limitations. Here, we employed Raman imaging for visualization and identification of polystyrene particles synthesized in polymerization reactions, ranging from 400 to 2600 nm. We successfully developed a quantitative model of particle size and concentration for polystyrene, exhibiting excellent fit (R2 of 0.9946). We established procedures for spiked flavored yogurt using synthesized polystyrene, providing fresh insights into microplastic extraction efficiency. Recovery rates calculated from models validated the method's feasibility. In practical applications, the assessment of the size, type, shape, and quantity of microplastics in unspiked flavored yogurt was conducted. The most common polymers found were polystyrene, polypropylene, and polyethylene, with the smallest polystyrene sizes ranging from 1 to 10 µm. Additionally, we conducted exposure assessments of microplastics in branded flavored yogurt. This study established a foundation for developing a universal method to quantify microplastics in food, covering synthesis of standards, method development, validation, and application.

Study on the Correlations between Quality Indicators of Dry-Aged Beef and Microbial Succession during Fermentation.

Dry-aged beef has been long favored by people due to its unique flavor and taste. However, the inner relationship between its overall quality formation and microbial changes during dry aging has not yet received much attention and research. To deeply reveal the forming mechanism of the unique flavor and taste of dry-aged beef, correlations between its three main quality indicators, i.e., texture, free amino acids (FAAs), volatile flavor compounds (VFCs), and microbial succession were analyzed in this study. The results showed that Staphylococcus spp. and Macrococcus spp. were key strains that influenced the total quality of dry-aged beef and strongly correlated with chewiness, hardness, and sweet FAAs (Ala), providing beef with unique palatability and taste. Additionally, among VFCs, Staphylococcus spp. and Macrococcus spp. showed a strong correlation with octanal and heptanal, and meanwhile, those highly correlated with nonanal, pentanol, and oct-1-en-3-ol were Debaryomyces spp., Psychrobacter spp., and Brochothrix spp., respectively, providing beef with a unique flavor. Staphylococcus spp. was proposed to be the dominant genus for dry-aged beef. This study provides valuable reference for the understanding of the role of microorganisms involved in dry aging.

Degradation of carbofuran and acetamiprid in wolfberry by dielectric barrier discharge plasma: Kinetics, pathways, toxicity and molecular dynamics simulation.

Carbofuran and acetamiprid pose the highest residual risk among pesticides found in wolfberries. This study aimed to degrade these pesticides in wolfberries using a multi-array dielectric barrier discharge plasma (DBD), evaluate the impact on safety and quality and explore their degradation mechanism. The results showed that DBD treatment achieved 90.6% and 80.9% degradation rates for carbofuran and acetamiprid, respectively, following a first-order kinetic reaction. The 120 s treatment successfully reduced pesticide contamination to levels below maximum residue limits. Treatment up to 180 s did not adversely affect the quality of wolfberries. QTOF/MS identification and degradation pathway analysis revealed that DBD broke down the furan ring and carbamate group of carbofuran, while replacing the chlorine atom and oxidizing the side chain of acetamiprid, leading to degradation. The toxicological evaluation showed that the degradation products were less toxic than undegraded pesticides. Molecular dynamics simulations revealed the reactive oxygen species (ROS) facilitated the degradation of pesticides through dehydrogenation and radical addition reactions. ROS type and dosage significantly affected the breakage of chemical bonds associated with toxicity (C4-O5 and C2-Cl1). These findings deepen insights into the plasma chemical degradation of pesticides.

Effect of tannic acid binding on the thermal degradation behavior and product toxicity of boscalid.

The effect of tannic acid (TA) binding on the thermal degradation of boscalid was studied in this work. The results revealed that TA binding has a significant impact on boscalid degradation. The degradation rate constant of bound boscalid was reduced, and its corresponding half-life was significantly prolonged compared to the free state. Four identical degradation products were detected in both states through UHPLC-Q-TOF-MS, indicating that degradation products were not affected by TA binding. Based on DFT and MS analysis, the degradation pathways of boscalid included hydroxyl substitution of chlorine atoms and cleavage of CN and CC bonds. The toxicity of B2 and B3 exceeded that of boscalid. In summary, the binding of TA and boscalid significantly affected the thermal degradation rate of boscalid while preserving the types of degradation products. This study contributed to a fundamental understanding of the degradation process of bound pesticide residues in complex food matrices.

The adsorption-desorption behavior of chlorothalonil in the cuticles of apple and red jujube.

The distribution fate of chlorothalonil (CHT) in the environment (soil and water) and fruits is controlled by the capacity of cuticles to adsorb and desorb CHT, which directly affects the safety of both the environment and fruits. Batch experiments were conducted to reveal the adsorption-desorption behaviors of CHT in the cuticles of apple and red jujube. The adsorption kinetics showed that both physisorption and chemisorption occurred during the adsorption process. Furthermore, the isothermal adsorption of CHT in the fruit cuticles followed the Freundlich model. The thermodynamic parameters (ΔG ≤ -26.16 kJ/mol, ΔH ≥ 31.05 kJ/mol, ΔS ≥ 0.20 kJ/(mol K) showed that the whole CHT adsorption process was spontaneous, and the hydrophobic interaction was predominant. The CHT adsorption capacity of the apple cuticle was higher than that of the red jujube cuticle, potentially due to the significantly higher alkanes content of apples than that of red jujubes. An appropriate ionic strength (0.01 moL/L) could induce a higher adsorption capacity. In addition, the desorption kinetics were shown to conform to a Quasi-first-order model, meaning that not all the adsorbed CHT could be easily desorbed. The desorption ratios in apple and red jujube cuticles were 41.38% and 35.64%, respectively. The results of Fourier-transformed infrared spectroscopy and X-ray photoelectron spectroscopy further confirmed that CHT could be adsorbed and retained in the fruit cuticles. Investigating the adsorption-desorption behavior of CHT in the apple and red jujube cuticles allowed to determine the ratio of its final distribution in the fruits and environment, providing a theoretical basis to evaluate the risk of residue pesticide.

Raman silent region - based method for detection of pesticides with cyano group.

Based on the fact that not all chemical substances possess good Raman signals, this article focuses on the Raman silent region signals of pesticides with cyano group. Under the optimized conditions of methanol-water (1:1, v/v) as the solvent, irradiation at 302 nm light source for 20 min, and the use of 0.5 mol/L KI as the aggregating agent, Surface-enhanced Raman spectroscopy (SERS) method for azoxystrobin detection was developed by the Raman silent region signal of 2230 cm-1, and verified by detecting the spiked grapes with different concentrations of azoxystrobin. Other four pesticides with cyano group also could be identified at the peak of 2180 cm-1, 2205 cm-1, 2125 cm-1, and 2130 cm-1 for acetamiprid, phoxim, thiacloprid and cymoxanil, respectively. When azoxystrobin or acetamiprid was mixed respectively with chlorpyrifos without cyano group, their SERS signals in the Raman silent region of chlorpyrifos were not interfered, while mixed with cymoxanil in different ratios (1:4, 1:1 and 4:1), respectively, each two pesticides with cyano group could be distinguished by the changes in the Raman silent region. In further, four pesticides with or without cyano group were mixed together in 1:1:1:1 (acetamiprid, cymoxanil, azoxystrobin chlorpyrifos), and each pesticide still could be identified even at 0.5 mg/L. The results showed that the SERS method combined with UV irradiation may provide a new way to monitor the pesticides with C≡N performance in the Raman silent region without interference from the food matrix.

Probing the effect of protein corona on SERS signals: insights from melamine detection in milk matrix.

Matrix effects limit the application of surface-enhanced Raman scattering (SERS) technology in the field of food safety. This study elucidated it from the perspective of protein corona by employing a model system for melamine SERS detection in milk. Compared with the melamine standard solution, higher detection limits (1 mg/L and 10 mg/L) are observed in milk matrix. The melamine signal exhibits an 80% reduction in whey protein solution, suggesting that protein has a significant impact on SERS signals. The changes in particle size, zeta potential and UV-vis spectra indicate the AuNPs interact with whey protein. Forming protein corona inhibits the melamine-induced AuNPs aggregation, reducing the number of 'hot spot' and the adsorption of melamine on AuNPs (from 0.28 mg/L to 0.07 mg/L), which may be responsible for signal loss. The found matrix effect from protein corona provides new insights for developing strategies about reducing matrix effect in SERS application.

First identification of microplastics in human uterine fibroids and myometrium.

Microplastics (MPs) pollution has received widespread attention in recent years as the use of plastics continues to increase. However, currently no studies have reported the finding of MPs in human uterine fibroids (UFs) and myometrium tissues. In this study, UFs tissues (n = 48) and myometrium tissues (n = 40) from 48 patients and myometrium tissues (n = 8) from healthy population were collected. Following digestion of the samples by 10% KOH and 30% H2O2, MPs were analyzed qualitatively and quantitatively using Raman spectroscopy. The 16 UFs and myometrium tissue samples contained an average of 1.5 ± 1.17 MP particles per gram of tissue. Notably, the abundance of MPs in the UFs tissues (2.13 ± 1.17 particles per gram) was higher than in the myometrium tissues (0.88 ± 0.78 particles per gram). In the same cohort of individuals with UFs, the quantities of MPs detected in the affected UFs tissue (2.63 ± 1.77 particles per gram) exceeded those detected in healthy tissue (1.08 ± 0.93 particles per gram), particularly in elderly patients. A correlation was observed between elevated MP levels and frequent consumption of takeout meals and bottled water among patients, indicating that MP ingestion through food sources might have contributed to the increased abundance and variety of MPs within UFs. Furthermore, UFs increased in size with higher concentrations of MPs, which may have been related to elevated levels of MPs-induced hormones. This study provides new insights into the assessment of the relationship between exposure to MPs and human disease risk.

Interaction behavior, mechanisms and hazardous changes of microplastics on single and binary component pesticide in the environment and food: Diethofencarb and pyrimethanil.

In order to investigate the adsorption behavior and mechanism of microplastics (MPs) on multiple coexisting pesticides in practical systems, as well as their hazardous changes upon binding, diethofencarb and pyrimethanil were selected to be studied with four MPs. The adsorption rate of both pesticides would be faster in the binary-component case, conforming to pseudo-second-order kinetics, with adsorption sites and chemical adsorption dominating. And the more hydrophobic the pesticide, the faster the adsorption rate and the higher the adsorption capacity. Diethofencarb belonged to monolayer adsorption, whereas pyrimethanil belonged to monomolecular combined with multilayer adsorption, depending on the size of pesticides. And the adsorption process was both competitive and synergistic when pesticides coexist. In addition, the adsorption process was a spontaneous heat absorption process. Electrostatic forces have little effect on adsorption, while the adsorption capacity can be altered by the adsorption sites and hydrophobicity of MPs. The salting-out effect also facilitated the adsorption process. As for changes in hazard, the bioluminescence of A. fischeri wasn't significantly inhibited, lacking of acute environmental toxicity. However, in vitro digestion experiments demonstrated a significant increase in bioavailability of diethofencarb and pyrimethanil in combination with MPs. These findings suggest the stronger adsorption behaviors and higher loading capacities between pesticides and MPs could lead more serious hazards to the human body, which deserves further attention.

First-line penpulimab combined with paclitaxel and carboplatin for metastatic squamous non-small-cell lung cancer in China (AK105-302): a multicentre, randomised, double-blind, placebo-controlled phase 3 clinical trial.

BACKGROUND: Penpulimab is a novel programmed death (PD)-1 inhibitor. This study aimed to establish the efficacy and safety of first line penpulimab plus chemotherapy for advanced squamous non-small-cell lung cancer. METHODS: This multicentre, randomised, double-blind, placebo-controlled, phase 3 clinical trial enrolled patients with locally advanced or metastatic squamous non-small-cell lung cancer from 74 hospitals in China. Eligible participants were aged 18-75 years, had histologically or cytologically confirmed locally advanced (stage IIIb or IIIc) or metastatic (stage IV) squamous non-small-cell lung cancer, were ineligible to complete surgical resection and concurrent or sequential chemoradiotherapy, had an Eastern Cooperative Oncology Group (ECOG) performance status of 0-1, did not have previous systemic chemotherapy for locally advanced or metastatic non-small-cell lung cancer, and had one or more measurable lesions according to RECIST (version 1.1). Participants were randomly assigned (1:1) to receive intravenous penpulimab 200 mg or placebo (excipient of penpulimab injection), plus paclitaxel 175 mg/m2 and carboplatin AUC of 5 intravenously on day 1 every 3 weeks for four cycles, followed by penpulimab or placebo as maintenance therapy. Stratification was done according to the PD-L1 tumour proportion score (<1% vs 1-49% vs ≥50%) and sex (male vs female). The participants, investigators, and other research staff were masked to group assignment. The primary outcome was progression-free survival assessed by the masked Independent Radiology Review Committee in the intention-to-treat population and patients with a PD-L1 tumour proportion score of 1% or more (PD-L1-positive subgroup). The primary analysis was based on the intention-to-treat analysis set (ie, all randomly assigned participants) and the PD-L1-positive subgroup. The safety analysis included all participants who received at least one dose of study drug after enrolment. This trial was registered with ClinicalTrials.gov (NCT03866993). FINDINGS: Between Dec 20, 2018, and Oct 10, 2020, 485 patients were screened, and 350 participants were randomly assigned (175 in the penpulimab group and 175 in the placebo group). Of 350 participants, 324 (93%) were male and 26 (7%) were female, and 347 (99%) were of Han ethnicity. In the final analysis (June 1, 2022; median follow-up, 24·7 months [IQR 0-41·4]), the penpulimab group showed an improved progression-free survival compared with the placebo group, both in the intention-to-treat population (median 7·6 months, 95% CI 6·8--9·6 vs 4·2 months, 95% CI 4·2-4·3; HR 0·43, 95% CI 0·33-0·56; p<0·0001) and in the PD-L1-positive subgroup (8·1 months, 5·7-9·7 vs 4·2 months, 4·1-4·3; HR 0·37, 0·27-0·52, p<0·0001). Grade 3 or worse treatment-emergent adverse events occurred in 120 (69%) 173 patients in the penpulimab group and 119 (68%) of 175 in the placebo group. INTERPRETATION: Penpulimab plus chemotherapy significantly improved progression-free survival in patients with advanced squamous non-small-cell lung cancer compared with chemotherapy alone. The treatment was safe and tolerable. Penpulimab combined with paclitaxel and carboplatin is a new option for first-line treatment in patients with this advanced disease. FUNDING: The National Natural Science Foundation of China, Shanghai Municipal Health Commission, Chia Tai Tianqing Pharmaceutical, Akeso.

Efficacy, Safety, and Population Pharmacokinetics of MW032 Compared With Denosumab for Solid Tumor-Related Bone Metastases: A Randomized, Double-Blind, Phase 3 Equivalence Trial.

Importance: The bioequivalence of denosumab biosimilar has yet to be studied in a 53-week, multicenter, large-scale, and head-to-head trial. A clinically effective biosimilar may help increase access to denosumab in patients with solid tumor-related bone metastases. Objectives: To establish the biosimilarity of MW032 to denosumab in patients with solid tumor-related bone metastases based on a large-scale head-to-head study. Design, Setting, and Participants: In this 53-week, randomized, double-blind, phase 3 equivalence trial, patients with solid tumors with bone metastasis were recruited from 46 clinical sites in China. Overall, 856 patients were screened and 708 eligible patients were randomly allocated to receive either MW032 or denosumab. Interventions: Patients were randomly assigned (1:1) to receive MW032 or reference denosumab subcutaneously every 4 weeks until week 49. Main Outcomes and Measures: The primary end point was percentage change from baseline to week 13 of natural logarithmic transformed urinary N-telopeptide/creatinine ratio (uNTx/uCr). Results: Among the 701 evaluable patients (350 in the MW032 group and 351 in the denosumab group), the mean (range) age was 56.1 (22.0-86.0) years and 460 patients were women (65.6%). The mean change of uNTx/uCr from baseline to week 13 was -72.0% (95% CI, -73.5% to -70.4%) in the MW032 group and -72.7% (95% CI, -74.2% to -71.2%) in the denosumab group. These percent changes corresponded to mean logarithmic ratios of -1.27 and -1.30, or a difference of 0.02. The 90% CI for the difference (-0.04 to 0.09) was within the equivalence margin (-0.13 to 0.13); the mean changes of uNTx/uCr and bone-specific alkaline phosphatase (s-BALP) at each time point were also similar during 53 weeks. The differences of uNTx/uCr change were 0.015 (95% CI, -0.06 to 0.09), -0.02 (95% CI, -0.09 to 0.06), -0.05 (95% CI, -0.13 to 0.03) and 0.001 (95% CI, -0.10 to 0.10) at weeks 5, 25, 37, and 53, respectively. The differences of s-BALP change were -0.006 (95% CI, 0.06 to 0.05), 0.00 (95% CI, -0.07 to 0.07), -0.085 (95% CI, -0.18 to 0.01), -0.09 (95% CI, -0.20 to 0.02), and -0.13 (95% CI, -0.27 to 0.004) at weeks 5, 13, 25, 37 and 53, respectively. No significant differences were observed in the incidence of skeletal-related events (-1.4%; 95% CI, -5.8% to 3.0%) or time to first on-study skeletal-related events (unadjusted HR, 0.86; P = .53; multiplicity adjusted HR, 0.87; P = .55) in the 2 groups. Conclusions and Relevance: MW032 and denosumab were biosimilar in efficacy, population pharmacokinetics, and safety profile. Availability of denosumab biosimilars may broaden the access to denosumab and reduce the drug burden for patients with advanced tumors. Trial Registration: ClinicalTrials.gov Identifier: NCT04812509.

Adipocytes-derived exosomal miR-122 promotes non-alcoholic fat liver disease progression via targeting Sirt1 / El miR-122 exosomal derivado de los adipocitos promueve la progresión de la enfermedad hepática por grasa no alcohólica a través de la focalización de Sirt1

Aims: Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease that affects adipose function. This study aimed to explore the function of adipocytes-derived exosomal (ADEs) miR-122 in NAFLD. Methods: A high-fat and high-fructose diet-induced rat model and a palmitic acid (PA)-induced in vitro model were established. The RNA level of miR-122 and Sirt1 was measured using qRT-PCR. The protein levels of exosome biomarkers, and lipogenesis, inflammation and fibrosis biomarkers were determined by western blotting. Cell viability and apoptosis were assessed using cell counting kit-8 and flow cytometry, respectively. Serum alanine aminotransferase, aspartate aminotransferase, total cholesterol, triglyceride levels were measured. Liver tissue damage was assessed using haematoxylin and eosin staining. The interaction between miR-122 and Sirt1 3′UTR was assessed using a luciferase reporter gene assay. Results: ADEs exhibited abundant level of miR-122 and promoted lipogenesis, impaired hepatocyte survival, enhanced liver damage and increased serum lipid levels in vivo and in vitro. Inhibition of miR-122 in ADEs alleviated NAFLD progression, lipid and glucose metabolism, liver inflammation and fibrosis both in vivo and in vitro. miR-122 binds directly to the 3′UTR of Sirt1 to suppress its expression. Moreover, Sirt1 overexpression reversed the increase in cell apoptosis, glucose and lipid metabolism, liver inflammation and fibrosis induced by ADEs in vivo and in vitro. Conclusions: The ADEs miR-122 promotes the progression of NAFLD via modulating Sirt1 signalling in vivo and in vitro. The ADEs miR-122 may be a promising diagnostic biomarker and therapeutic target for NAFLD.(AU)

Objetivos: La enfermedad del hígado graso no alcohólico (EHGNA) es una enfermedad hepática crónica que afecta a la función adiposa. Este estudio tiene como objetivo explorar la función de los exosomas derivados de los adipocitos (ADEs) miR-122 en la EHGNA. Métodos: Se estableció un modelo de rata inducido por una dieta alta en grasas y fructosa y un modelo in vitro inducido por ácido palmítico (AP). Se midió el nivel de ARN de miR-122 y Sirt1 mediante qRT-PCR. Los niveles de proteína de los biomarcadores de exosomas y los biomarcadores de lipogénesis, inflamación y fibrosis se determinaron mediante western blotting. La viabilidad celular y la apoptosis se evaluaron mediante el kit de recuento de células-8 y la citometría de flujo, respectivamente. Se midieron los niveles séricos de alanina aminotransferasa, aspartato aminotransferasa, colesterol total y triglicéridos. El daño tisular del hígado se evaluó mediante tinción con hematoxilina y eosina. La interacción entre miR-122 y Sirt1 3’UTR se evaluó mediante un ensayo de gen reportero de luciferasa. Resultados: Los ADEs mostraron un nivel abundante de miR-122 y promovieron la lipogénesis, perjudicaron la supervivencia de los hepatocitos, potenciaron el daño hepático y aumentaron los niveles de lípidos séricos in vivo e in vitro. La inhibición de miR-122 en los ADEs alivió la progresión de la EHGNA, el metabolismo de los lípidos y la glucosa, la inflamación del hígado y la fibrosis tanto in vivo como in vitro. miR-122 se une directamente a la 3’UTR de Sirt1 para suprimir su expresión. Además, la sobreexpresión de Sirt1 revirtió el aumento de la apoptosis celular, el metabolismo de la glucosa y los lípidos, la inflamación del hígado y la fibrosis inducida por los ADEs in vivo e in vitro. Conclusiones: El ADEs miR-122 promueve la progresión de la EHGNA a través de la modulación de la señalización de Sirt1 in vivo e in vitro...(AU)