A novel selective ERK1/2 inhibitor, Laxiflorin B, targets EGFR mutation subtypes in non-small-cell lung cancer.

Extracellular regulated protein kinases 1/2 (ERK1/2) are key members of multiple signaling pathways, including the ErbB axis. Ectopic ERK1/2 activation contributes to various types of cancer, especially drug resistance to inhibitors of RTK, RAF and MEK, and specific ERK1/2 inhibitors are scarce. In this study, we identified a potential novel covalent ERK inhibitor, Laxiflorin B, which is a herbal compound with anticancer activity. However, Laxiflorin B is present at low levels in herbs; therefore, we adopted a semi-synthetic process for the efficient production of Laxiflorin B to improve the yield. Laxiflorin B induced mitochondria-mediated apoptosis via BAD activation in non-small-cell lung cancer (NSCLC) cells, especially in EGFR mutant subtypes. Transcriptomic analysis suggested that Laxiflorin B inhibits amphiregulin (AREG) and epiregulin (EREG) expression through ERK inhibition, and suppressed the activation of their receptors, ErbBs, via a positive feedback loop. Moreover, mass spectrometry analysis combined with computer simulation revealed that Laxiflorin B binds covalently to Cys-183 in the ATP-binding pocket of ERK1 via the D-ring, and Cys-178 of ERK1 through non-inhibitory binding of the A-ring. In a NSCLC tumor xenograft model in nude mice, Laxiflorin B also exhibited strong tumor suppressive effects with low toxicity and AREG and EREG were identified as biomarkers of Laxiflorin B efficacy. Finally, Laxiflorin B-4, a C-6 analog of Laxiflorin B, exhibited higher binding affinity for ERK1/2 and stronger tumor suppression. These findings provide a new approach to tumor inhibition using natural anticancer compounds.

Magnetic resonance imaging-based Classifications for Symptom of Adenomyosis.

OBJECTIVES: To identify an optimal magnetic resonance imaging (MRI)-based classification for the severity of adenomyosis and explore the factors associated with disease severity (dysmenorrhea or menorrhagia). DESIGN AND PARTICIPANTS: Several classifications based on MRI have been proposed, and their phenotypes are reported to be associated with the severity of adenomyosis. However, a consensus classification based on MRI findings has not yet been reached. Our study was designed to retrospectively analyze data from a cohort of patients in the Affiliated Nanchong Central Hospital of North Sichuan Medical College from June 2017 to December 2021 before focused ultrasound ablation surgery (FUAS), to identify the optimal classification of adenomyosis severity from different classification criteria and explore factors associated with the presence of symptoms. METHODS: The proportions of disease severity among different classification groups were compared to obtain the one generating the most considerable chi-square value, which was identified as the optimal classification for informing disease severity. A logistic regression model was constructed to explore factors associated with disease severity. RESULTS: Classification of Kobayashi H (classification 4) concerning the affected areas and size (volumes of lesions) was recognized as the optimal one, which identified dysmenorrhea (χ2=18.550, p-value=0.002) and menorrhagia (χ2=15.060, p-value=0.010) secondary to adenomyosis. For volumes of uterine wall <2/3, the dysmenorrhea rate in subtype-4 was higher than that in subtype-1 (χ2=4.114, p-value=0.043), and the dysmenorrhea rate in subtype-5 was higher than that in subtype-2 (χ2=4.357, p-value=0.037). Age (OR=0.899, 95%CI=0.810～0.997, p-value=0.044) and external phenotype (OR=3.588, 95%CI=CI 1.018～12.643, p-value=0.047) were associated with dysmenorrhea. Concerning volumes of uterine wall ≥2/3, the menorrhagia rate in subtype-3 remarkably increased compared with that in subtype-6 (χ2=9.776, p-value=0.002), and internal phenotype was identified as an independent factor associated with menorrhagia (OR=1.706, 95%CI=1.131～2.573, p-value=0.011). LIMITATIONS: Patients in our study were all included before FUAS, which limited our result interpretation for the general patient population. CONCLUSIONS: MRI-based classification 4 is identified as an optimal classification for informing the severity of adenomyosis. The phenotype of classification is the main characteristic associated with disease severity.

Coagulation after paediatric miniaturised versus conventional cardiopulmonary bypass: Retrospective cohort study.

BACKGROUND: Cardiopulmonary bypass (CPB) causes coagulation disorders after surgery. This study aimed to compare the coagulation parameters after congenital cardiac surgery with miniaturised CPB (MCPB) versus conventional CPB (CCPB). METHODS: We gathered information about children who underwent cardiac surgery between 1/1/2016 and 12/31/2019. Using propensity score-matched data, we compared the coagulation parameters and postoperative outcomes of the MCPB and CCPB groups. RESULTS: A total of 496 patients (327 with MCPB, 169 withCCPB) underwent congenital cardiac surgery, and 160 matched pairs in each group were enrolled in the analysis. Compared with CCPB children, MCPB children had a lowermean prothrombin time (14.9 ± 2.0 vs 16.4 ± 4.1; p < 0.001)and international normalised ratio (1.3 ± 0.2 vs. 1.4 ± 0.3; p < 0.001), but higher thrombin time (23.4 ± 20.4 vs 18.2 ± 4.4; p = 0.002). The CCPB group had greaterperioperative changes inprothrombin time, international normalised ratio, fibrinogen, and antithrombin III activity (all p < 0.01) but lower perioperative changesin thrombin time (p = 0.001) thanthe MCPB group. Ultra-fasttrack extubation and blood transfusionrates, postoperative blood loss, and intensive care unitlength of stay were considerably decreased in the MCPB group. There were no considerable intergroup differences in the activated partial thromboplastin time or platelet count. CONCLUSIONS: Compared with CCPB, MCPB was associated with lower coagulation changes and better early outcomes, including shorter intensive care unit stay and less postoperative blood loss.

High internal phase Pickering emulsions stabilised by ultrasound-induced soy protein-ß-glucan-catechin complex nanoparticles to enhance the stability and bioaccessibility of curcumin.

AIMS: To evaluate the potential applications of soy protein-glucan-catechin (SGC) complexes prepared with different ultrasound times in stabilising high internal phase Pickering emulsion (HIPPE) and delivering curcumin. METHODS: The SGC complexes were characterised by particle size, morphology, zeta potential, Fourier transform infra-red, and fluorescence spectroscopy. Formation and stability of curcumin emulsions were monitored by droplet size, microstructure, rheological property, lipid oxidation, and in vitro digestion. RESULTS: Short-time ultrasound-induced complexes (SGC-U15) exhibited a small size and wettability of ∼82.5°. The chemical stability and bioaccessibility of curcumin was greatly improved by SGC-U15-stabilised HIPPEs, even after 70 days of storage, heating at 100 °C for 30 min, ultraviolet irradiation for 120 min, and in vitro digestion, owing to the formation of elastic gel-like structure at the oil/water interfaces. CONCLUSION: Our findings may contribute to the design of emulsion-based delivery systems using ultrasound-induced protein-polysaccharide-polyphenol complexes.

Dietary Proteins Alter Fermentation Characteristics of Human Gut Microbiota In Vitro.

The objective of this study was to evaluate the fermentation characteristics of proteins from diverse sources by human gut microbiota. Cereal proteins (rice and oat), red meat proteins (pork and beef), chicken protein and casein were selected as the substrates for simulated gastrointestinal digestion (SGID), and human faecal samples were collected from healthy donors as the inoculum of fermentation. In this study, we further analyzed the correlations of amino acids (AA) compositions, fermentation productions and gut microbiota. As the results, the animal protein groups had higher degree of hydrolysis (DH) after digestion and higher levels of ammonia nitrogen (NH3-N) after fermentation than cereal proteins. The pH value of fermentation liquid declined as proteins were added during fermentation. Cereal protein groups promoted the gut microbiota to produce more short chain fatty acids (SCFAs) with the high proportion of acetate, propionate and butyrate by lowering the pH than red meat proteins. The abundance of Firmicutes at phylum level in cereal protein groups was lower than red meat proteins after fermentation. The cereal protein groups enhanced the growth of Bacteroides spp. and Bifidobacterium spp. while red meat proteins stimulated the growth of Peptoclostridium spp.. Taken together, our research implies that cereal proteins have better fermentation characters than red meat proteins.

Simultaneous determination of dyes in wines by HPLC coupled to quadrupole orbitrap mass spectrometry.

A new method combining the QuEChERS (quick, easy, cheap, effective, rugged, and safe) method with ultra high performance liquid chromatography and ESI quadrupole Orbitrap high-resolution MS was developed for the highly accurate and sensitive screening of 69 dyes in wines. Response surface methodology was employed to optimize the QuEChERS sample preparation method for the determination of 69 different analytes in wines for the first time. After optimization, the maximum predicted recovery was 99.48% rate for canacert indigo carmine under the optimized conditions of 10 mL acetonitrile, 1.45 g sodium acetate, 107 mg primary secondary amine, and 96 mg C18 . For the matrices studied, the recovery rates of the other 68 compounds ranged from 87.2-107.4%, with coefficient of variation < 6.4%. The mass accuracy typically obtained is routinely better than 1.6 ppm and only needed to be calibrated once a week. The LODs for the analytes are in the range 1-1000 µg/kg. This method has been successfully applied on screening of dyes in commercial wines, and it is very useful for the fast screening of different food additives.

A biomimetic synthesis of (±)-basiliolide B.

A highly diastereoselective and practical biomimetic total synthesis of (±)-basiliolide B has been achieved through the study of the two proposed biosynthetic pathways (O-methylation and O-acylation) for the unprecedented 7-methoxy-4,5-dihydro-3H-oxepin-2-one (C ring). The synthesis featured a cyclopropanation/ring opening strategy for establishing the stereogenic centers at C8 and C9, a biomimetic 2-pyrone Diels-Alder cycloaddition for the synthesis of the ABD ring system, and finally a highly efficient biomimetic intramolecular O-acylation for the C ring formation. This result provides an important perspective on the biosynthetic origin of the unprecedented 7-membered acyl ketene acetal moiety of the C ring.

Advanced ethylene-absorbing and cushioning depending on the 3D porous-structured fruit packaging: Toward polyurethane foam manipulation using zein and soybean oil polyol substrates.

Fruits are essential sources of nutrients in our daily diet; however, their spoilage is often intensified by mechanical damage and the ethylene phytohormone, resulting in significant economic losses and exacerbating hunger issues. To address these challenges, this study presented a straightforward in situ synthesis protocol for producing Z/SOPPU foam, a 3D porous-structured fruit packaging. This innovative packaging material offered advanced ethylene-adsorbing and cushioning capabilities achieved through stirring, heating, and standing treatments. The results demonstrated that the Z/SOPPU foam, with its porous structure, served as an excellent packaging material for fruits, maintaining the intact appearance of tomatoes even after being thrown 72 times from a height of 1.5 m. Additionally, it exhibited desirable hydrophobicity (contact angle of 114.31 ± 0.82°), degradability (2.73 ± 0.88 % per 4 weeks), and efficient ethylene adsorption (adsorption rate of 13.2 ± 1.7 mg/m3/h). These remarkable characteristics could be attributed to the unique 3D micron-porous configuration, consisting of soybean oil polyol polyurethane foam for mechanical strain cushioning and zein for enhanced ethylene adsorption efficiency. Overall, this research offers an effective and original approach to the rational design and fabrication of advanced bio-based fruit packaging.

Recent advances and applications in high-throughput continuous flow.

High-throughput continuous flow technology has emerged as a revolutionary approach in chemical synthesis, offering accelerated experimentation and improved efficiency. With the aid of process analytical technology and automation, this system not only enables rapid optimisation of reaction conditions at the millimole to the picomole scale, but also facilitates automated scale-up synthesis. It can even achieve the self-planning and self-synthesis of small drug molecules with artificial intelligence incorporated in the system. The versatility of the system is highlighted by its compatibility with both electrochemistry and photochemistry, and its significant applications in organic synthesis and drug discovery. This highlight summarises its recent developments and applications, emphasising its significant impact on advancing research across multiple disciplines.

Effects of soaking glutinous sorghum grains on physicochemical properties of starch.

Glutinous sorghum grains were soaked (60-80 °C, 2-8 h) to explore the effects of soaking, an essential step in industrial processing of brewing, on starch. As the soaking temperature increased, the peak viscosity and crystallinity of starch gradually decreased, while the enzymatic hydrolysis rate and storage modulus first increased and then decreased. At 70 °C, the content of amylose, the enzymatic hydrolysis rate of starch, and the final viscosity first increase and then decrease with the increase of soaking time, reaching their maximum at 6 h, increased by 53.1 %, 11.0 %, and 10.4 %, respectively, as compared with the non-soaked sample. At 80 °C (4 h), the laser confocal microscopy images showed a network structure formed between the denatured protein chains and the leached-out amylose chains. The molecular weights of starch before and after soaking were all in the range of 3.82-8.98 × 107 g/mol. Since 70 °C is lower than that of starch gelatinization and protein denaturation, when soaking for 6 h, the enzymatic hydrolysis rate of starch is the highest, and the growth of miscellaneous bacteria is inhibited, which is beneficial for subsequent processing technology. The result provides a theoretical basis for the intelligent control of glutinous sorghum brewing.

Copper-catalyzed asymmetric allylic substitution of racemic/meso substrates.

The synthesis of enantiomerically pure compounds is a pivotal subject in the field of chemistry, with enantioselective catalysis currently standing as the primary approach for delivering specific enantiomers. Among these strategies, Cu-catalyzed asymmetric allylic substitution (AAS) is significant and irreplaceable, especially when it comes to the use of non-stabilized nucleophiles (pK a > 25). Although Cu-catalyzed AAS of prochiral substrates has also been widely developed, methodologies involving racemic/meso substrates are highly desirable, as the substrates undergo dynamic processes to give single enantiomer products. Inspired by the pioneering work of the Alexakis, Feringa and Gennari groups, Cu-catalyzed AAS has been continuously employed in deracemization and desymmetrization processes for the synthesis of enantiomerically enriched products. In this review, we mainly focus on the developments of Cu-catalyzed AAS with racemic/meso substrates over the past two decades, providing an explicit outline of the ligands employed, the scope of nucleophiles, the underlying dynamic processes and their practical applications.

Desymmetrization of Inert meso-Diethers through Copper-Catalyzed Asymmetric Allylic Alkylation with Grignard Reagents.

We have developed a highly regio-, diastereo-, and enantioselective Cu-catalyzed desymmetrization of inert meso-diethers using Grignard reagents. Moreover, previous inaccessible sterically hindered organometallic reagents are realized in the reaction with broad secondary alkyl Grignard reagents. Finally, detailed control experiments and density functional theory calculations revealed the desymmetrization of meso-diethers exploits a direct anti-SN2' pathway, in the absence of an in situ-generated allyl bromine intermediate. The following oxidative addition is the crucial rate-determining and enantioselectivity-determining step.

Distinct effects of phyllosphere and rhizosphere microbes on invader Ageratina adenophora during its early life stages.

Microbes strongly affect invasive plant growth. However, how phyllosphere and rhizosphere soil microbes distinctively affect seedling mortality and growth of invaders across ontogeny under varying soil nutrient levels remains unclear. In this study, we used the invader Ageratina adenophora to evaluate these effects. We found that higher proportions of potential pathogens were detected in core microbial taxa in leaf litter than rhizosphere soil and thus leaf inoculation had more adverse effects on seed germination and seedling survival than soil inoculation. Microbial inoculation at different growth stages altered the microbial community and functions of seedlings, and earlier inoculation had a more adverse effect on seedling survival and growth. The soil nutrient level did not affect microbe-mediated seedling growth and the relative abundance of the microbial community and functions involved in seedling growth. The effects of some microbial genera on seedling survival are distinct from those on growth. Moreover, the A. adenophora seedling-killing effects of fungal strains isolated from dead seedlings by non-sterile leaf inoculation exhibited significant phylogenetic signals, by which strains of Allophoma and Alternaria generally caused high seedling mortality. Our study stresses the essential role of A. adenophora litter microbes in population establishment by regulating seedling density and growth.

The effects of resonance acoustic mixing modulation on the structural and emulsifying properties of pea protein isolate.

The effects of resonant acoustic mixing (RAM) with different treatment times (0, 5, 10, 15, 20 and 30 min) on the structural and emulsifying properties of pea protein isolate (PPI) were investigated for the first time. Increasing the RAM treatment time from 0 to 20 min decreased the α-helix/ß-sheet ratio and particle size of the PPI samples by 37.84 % and 46.44 %, respectively, accompanied by an increase in solubility from 54.79 % to 71.80 % (P < 0.05). Consequently, the emulsifying activity index of PPI (from 10.45 m2/g to 14.2 m2/g) and the physical stability of RAM-PPI emulsions were effectively enhanced, which was confirmed by the small and uniformly distributed oil droplets in the micrographs of the emulsions. However, excessive RAM treatment (30 min) diminished the effectiveness of the aforementioned improvements. Therefore, obviously enhanced solubility and emulsifying properties of PPI can be attained through proper RAM treatment (15-20 min).

Neferine inhibits BMECs pyroptosis and maintains blood-brain barrier integrity in ischemic stroke by triggering a cascade reaction of PGC-1α.

Blood-brain barrier disruption is a critical pathological event in the progression of ischemic stroke (IS). Most studies regarding the therapeutic potential of neferine (Nef) on IS have focused on neuroprotective effect. However, whether Nef attenuates BBB disruption during IS is unclear. We here used mice underwent transient middle cerebral artery occlusion (tMCAO) in vivo and bEnd.3 cells exposed to oxygen-glucose deprivation/reoxygenation (OGD/R) injury in vitro to simulate cerebral ischemia. We showed that Nef reduced neurobehavioral dysfunction and protected brain microvascular endothelial cells and BBB integrity. Molecular docking, short interfering (Si) RNA and plasmid transfection results showed us that PGC-1α was the most binding affinity of biological activity protein for Nef. And verification experiments were showed that Nef upregulated PGC-1α expression to reduce mitochondrial oxidative stress and promote TJ proteins expression, further improves the integrity of BBB in mice. Intriguingly, our study showed that neferine is a natural PGC-1α activator and illustrated the mechanism of specific binding site. Furthermore, we have demonstrated Nef reduced mitochondria oxidative damage and ameliorates endothelial inflammation by inhibiting pyroptosis to improve BBB permeability through triggering a cascade reaction of PGC-1α via regulation of PGC-1α/NLRP3/GSDMD signaling pathway to maintain the integrity of BBB in ischemia/reperfusion injury.

Inhibition of Microglial Activation by Delayed Mild Hypothermia Reduced Preoligodendrocyte Injury in a Neonatal Rat Brain Slice Model.

Periventricular leukomalacia (PVL), characterized by distinctive form of white matter injury, often arises after neonatal cardiac surgery. Proven therapies for PVL are absent. In this study, we designed to quest therapeutic effects of delayed mild hypothermia on PVL and its mechanism in a neonatal rat brain slice model. With the increase of delayed mild hypothermia-treating time, the reduced expression of myelin basic protein and loss of preoligodendrocytes were significantly attenuated after oxygen-glucose deprivation. In addition, the proportion of ionized calcium binding adapter molecule 1 (Iba-1)-positive cells and the expression of Iba-1 were apparently reduced with the increased duration of mild hypothermia treatment. Furthermore, the levels of tumor necrosis factor alpha and interleukin-6 reduced after the mild hypothermia treatment relative to the control. Inhibition of microglial activation with prolonged mild hypothermia may be a potential strategy for white matter protection during cardiopulmonary bypass and hypothermic circulatory arrest.

Soy protein/ß-glucan/tannic acid complex coacervates with different micro-structures play key roles in the rheological properties, tribological properties, and the storage stability of Pickering high internal phase emulsions.

Soy protein-glucan-tannic acid (S-G-T) complex coacervates were prepared by high-intensity ultrasonic processing (US), subcritical water (SW) treatment, and microfluidization homogenization with different pressures (DHPM 500 and 1000) to stabilize Pickering high internal phase emulsions (HIPEs). Our results show that different treatments affect the microstructure and interfacial properties of ternary coacervates by modifying the intermolecular associations between components. In particular, US and DHPM 500 resulted in the formation of ternary complexes with smaller particle sizes and compact network structures. These properties facilitate the capture of oil droplets. S-G-T (US)- and S-G-T (DHPM500)-stabilized HIPEs exhibit gel-like structures, good creaming stability, and high friction coefficients due to the formation of a three-dimensional network in the continuous phase. However, a more uniform droplet distribution, higher viscoelasticity, and reduced friction were observed after storage for 60 days. These results provide a strategy to improve the sensory characteristics of emulsion-based foods after processing.

Molecular mechanisms affecting the stability of high internal phase emulsions of zein-soy isoflavone complexes fabricated with ultrasound-assisted dynamic high-pressure microfluidization.

In this study, zein-soy isoflavone complex (ZSI) emulsifiers were fabricated using ultrasound-assisted dynamic high-pressure micro fluidization to stabilise highinternal phase pickering emulsions. Ultrasound-assisted dynamic high-pressure micro-fluidization enhanced surface hydrophobicity, zeta potential, and soy isoflavone binding capacity, while it decreased particle size, especially during ultrasound and subsequent microfluidization. The treated ZSI could produce small droplet clusters and gel-like structures, with excellent viscoelasticity, thixotropy and creaming stability owing to their neutral contact angles. Ultrasound and subsequent micro fluidization treatment of the ZSI complexes were highly effective in preventing droplet flocculation and coalescence after long-term storage or centrifugation due to their higher surface load, thicker multi-layer interfacial structure, and stronger electronic repulsion between the oil droplets. This study provides insights and extends our current knowledge of how non-thermal technology affects the interfacial distribution of plant based particles and the physical stability of emulsions.

Neuroprotective Effects of a Novel Tetrapeptide SGGY from Walnut against H2O2-Stimulated Oxidative Stress in SH-SY5Y Cells: Possible Involved JNK, p38 and Nrf2 Signaling Pathways.

SGGY, an antioxidant tetrapeptide identified from walnut protein hydrolysate in our previous study, has been suggested to possess the potential to alleviate oxidative stress in cells. In this paper, the neuroprotective effects of SGGY on H2O2-stimulated oxidative stress in SH-SY5Y cells and the underlying mechanisms were investigated. Results showed that SGGY alleviated H2O2-induced oxidative stress by decreasing the intracellular reactive oxygen species (ROS) level and altering the mitochondrial membrane potential (MMP), thereby inhibiting apoptosis and increasing cell viability. SGGY significantly restored antioxidant enzyme activities and reduced malondialdehyde (MDA) content accordingly. Moreover, SGGY promoted the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) and suppressed the H2O2-induced activation of JNK and p38 mitogen-activated protein kinases (MAPKs). Taken together, these results suggested that SGGY protected SH-SY5Y cells from H2O2-provoked oxidative stress by enhancing the ability of cellular antioxidant defense, and the possible mechanism involved MAPKs and Nrf2 signaling pathways.