Ruscogenin Exerts Anxiolytic-Like Effect via Microglial NF-κB/MAPKs/NLRP3 Signaling Pathways in Mouse Model of Chronic Inflammatory Pain.

Long-term inflammation can cause chronic pain and trigger patients' anxiety by sensitizing the central nervous system. However, effective drugs with few side effects for treating chronic pain-induced anxiety are still lacking. The anxiolytic and anti-inflammatory effects of ruscogenin (RUS), an important active compound in Ophiopogon japonicus, were evaluated in a mouse model of chronic inflammatory pain and N9 cells. RUS (5, 10, or 20 mg/kg/day, i.g.) was administered once daily for 7 days after CFA injection; pain- and anxiety-like behaviors were assessed in mice. Anti-inflammatory effect of RUS (0.1, 1, 10 µM) on N9 microglia after LPS treatment was evaluated. Inflammatory markers (TNF-α, IL-1ß, IL-6, CD86, IL-4, ARG-1, and CD206) were measured using qPCR. The levels of IBA1, ROS, NF-κB, TLR4, P-IKK, P-IκBα, and P65, MAPKs (ERK, JNK, and P38), NLRP3 (caspase-1, ASC, and NLRP3) were detected by Western blotting or immunofluorescence staining. The potential target of RUS was validated by molecular docking and adeno-associated virus injection. Mice in CFA group exhibited allodynia and anxiety-like behaviors. LPS induced neuroinflammation in N9 cells. Both CFA and LPS increased the levels of IBA1, ROS, and inflammatory markers. RUS (10 mg/kg in vivo and 1 µM in vitro) alleviated these alterations through NF-κB/MAPKs/NLRP3 signaling pathways but had no effect on pain hypersensitivity. TLR4 strongly interacted with RUS, and TLR4 overexpression abolished the effects of RUS on anxiety and neuroinflammation. RUS exerts anti-inflammatory and anxiolytic effects via TLR4-mediated NF-κB/MAPKs/NLRP3 signaling pathways, which provides a basis for the treatment of chronic pain-induced anxiety.

Advanced application of slightly acidic electrolyzed water for fresh-cut fruits and vegetables preservation.

Fresh-cut fruits and vegetables (F&V) play a pivotal role in modern diets due to their convenience and nutritional value. However, their perishable nature renders them susceptible to rapid spoilage, causing quality deterioration, safety risks, and economic losses along the supply chain. Traditional preservation methods, while effective to some extent, often fall short in maintaining the quality and safety of fresh-cut F&V. This comprehensive review examines the utilization of slightly acidic electrolyzed water (SAEW) as a novel preservation technique for fresh-cut F&V. The review encompasses the production mechanisms, sterilization principles, classifications and application of SAEW. It explores the effects of SAEW on microbial inactivation, quality parameters, and metabolic pathways in fresh-cut F&V. Additionally, it assesses the synergistic effects of SAEW when combined with other preservation methods. SAEW demonstrates remarkable potential in extending the shelf life of fresh-cut F&V by effectively inhibiting microbial growth, suppressing browning, preserving chemical content, and influencing various metabolic processes. Moreover, its synergy with different treatments enhances its overall efficacy in maintaining fresh-cut F&V quality. The review highlights the promising role of SAEW as an innovative preservation approach for fresh-cut F&V. However, challenges regarding its widespread implementation and potential limitations require further exploration. Overall, SAEW stands as a significant contender in ensuring the safety and quality of fresh-cut F&V paving the way for future research and application in the food industry.

L-Cysteine: A promising nutritional supplement for alleviating anxiety disorders.

Anxiety disorders are prevalent chronic psychological disease with complex pathogenic mechanisms. Current anxiolytics have limited efficacy and numerous side effects in many anxiety patients, highlighting the urgent need for new therapies. Recent research has been focusing on nutritional supplements, particularly amino acids, as potential therapies for anxiety disorders. Among these, L-Cysteine plays a crucial role in various biological processes. L-Cysteine exhibits antioxidant properties that can enhance the antioxidant functions of the central nervous system (CNS). Furthermore, metabolites of L-cysteine, such as glutathione and hydrogen sulfide have been shown to alleviate anxiety through distinct molecular mechanisms. Long-term administration of L-Cysteine has anxiolytic, antidepressant, and memory-improving effects. L-Cysteine depletion can lead to increased oxidative stress in the brain. This review delves into the potential mechanisms of L-Cysteine and its main products, glutathione (GSH) and hydrogen sulfide (H2S) in the management of anxiety and related diseases.

Enhancing Fresh-Cut Apple Preservation: Impact of Slightly Acidic Electrolyzed Water and Chitosan-Apple Essence Microencapsulation Coating on Browning and Flavor.

Fresh-cut apple preservation is a critical concern in the food industry due to the rapid deterioration of texture, color, and flavor. While our previous study introduced apple essence microencapsulation (AEM) to enhance flavor during storage, its impact on overall storage quality was minimal. Thus, this study explores the application of two preservation techniques, namely, slightly acidic electrolyzed water (SAEW) and chitosan-apple essence microencapsulation (CH-AEM) coating, to enhance the quality of fresh-cut apples. Our findings reveal that SAEW treatment significantly reduces the browning index (from 65.38 to 57.36) and respiratory rate (from 5.10% to 4.30% of CO2), and maintains a desirable aroma profile compared to uncoated treatment during 10 days of storage. Additionally, the CH-AEM coating acts as a protective barrier, further preserving the sensory characteristics of fresh-cut apples. Notably, the SAEW-CH-AEM group exhibits superior performance in firmness (8.14 N), respiratory rate (3.37% of CO2), ion leakage (34.86%), and juice yield (47.52%) after 10 days. Our research highlights the synergistic effect of combining these preservation strategies, providing a promising approach for extending the shelf life of fresh-cut apples while maintaining their visual appeal and aromatic quality. These results offer valuable insights for the fresh-cut produce industry, contributing to improved apple product preservation and consumer satisfaction.

Mice harboring the T316N variant in the GABAAR γ2 subunit exhibit sleep-related hypermotor epilepsy phenotypes and hypersynchronization in the thalamocortical pathway.

OBJECTIVE: Sleep-related hypermotor epilepsy (SHE) is a focal epilepsy syndrome characterized by seizures that predominantly occur during sleep. The pathogenesis of these seizures remains unclear. We previously detected rare variants in GABRG2, which encodes the γ2 subunit of γ-aminobutyric acid type A receptor (GABAAR), in patients with SHE and demonstrated that these variants impaired GABAAR function in vitro. However, the mechanisms by which GABRG2 variants contribute to seizure attacks during sleep remain unclear. METHODS: In this study, we designed a knock-in (KI) mouse expressing the mouse Gabrg2 T316N variant, corresponding to human GABRG2 T317N variant, using CRISPR/Cas9. Continuous video-electroencephalogram monitoring and in vivo multichannel electrophysiological recordings were performed to explore seizure susceptibility to pentylenetetrazol (PTZ), alterations in the sleep-wake cycle, spontaneous seizure patterns, and synchronized activity in the motor thalamic nuclei (MoTN) and secondary motor cortex (M2). Circadian variations in the expression of total, membrane-bound, and synaptic GABAAR subunits were also investigated. RESULTS: No obvious changes in gross morphology were detected in Gabrg2T316N/+ mice compared to their wild-type (Gabrg2+/+) littermates. Gabrg2T316N/+ mice share key phenotypes with patients, including sleep fragmentation and spontaneous seizures during sleep. Gabrg2T316N/+ mice showed increased susceptibility to PTZ-induced seizures and higher mortality after seizures. Synchronization of the local field potentials between the MoTN and M2 was abnormally enhanced in Gabrg2T316N/+ mice during light phase, when sleep dominates, accompanied by increased local activities in the MoTN and M2. Interestingly, in Gabrg2+/+ mice, GABAAR γ2 subunits showed a circadian increase on the neuronal membrane and synaptosomes in the transition from dark phase to light phase, which was absent in Gabrg2T316N/+ mice. CONCLUSION: We generated a new SHE mouse model and provided in vivo evidence that rare variants of GABRG2 contribute to seizure attacks during sleep in SHE.

Inactivation mechanisms on pectin methylesterase by high pressure processing combined with its recombinant inhibitor.

High pressure processing (HPP) juice often experiences cloud loss during storage, caused by the activity of pectin methylesterase (PME). The combination of HPP with natural pectin methylesterase inhibitor (PMEI) could improve juice stability. However, extracting natural PMEI is challenging. Gene recombination technology offers a solution by efficiently expressing recombinant PMEI from Escherichia coli and Pichia pastoris. Experimental and molecular dynamics simulation were conducted to investigate changes in activity, structure, and interaction of PME and recombinant PMEI during HPP. The results showed PME retained high residual activity, while PMEI demonstrated superior pressure resistance. Under HPP, PMEI's structure remained stable, while the N-terminus of PME's α-helix became unstable. Additionally, the helix at the junction with the PME/PMEI complex changed, thereby affecting its binding. Furthermore, PMEI competed with pectin for active sites on PME, elucidating. The potential mechanism of PME inactivation through the synergistic effects of HPP and PMEI.

TOM40 mediates the effect of TSPO on postpartum depression partially through regulating calcium homeostasis in microglia.

AIMS: To assess the effect of the translocator protein 18 kDa (TSPO) on postpartum depression and explore its mechanism. METHODS: Postpartum depression (PPD) mouse model was established, and flow cytometry, immunofluorescence, Western blot analysis, real-time quantitative PCR, adeno-associated virus (AAV), co-immunoprecipitation-mass spectrometry and immunofluorescence co-staining were used to detect the effect of TSPO ligand ZBD-2 on PPD mice. RESULTS: ZBD-2 inhibits the overactivation of microglia in the hippocampus and amygdala of PPD model mice. ZBD-2 not only inhibited the inflammation but also repressed the burst of reactive oxygen species (ROS) and mitochondrial ROS (mtROS). Meanwhile, ZBD-2 protects mitochondria from LPS-induced damages through inhibiting the influx of calcium. ZBD-2 modulated the calcium influx by increasing the level of translocase of the outer mitochondrial membrane 40 (TOM40) and reducing the interaction of TSPO and TOM40. In addition, the effect of ZBD-2 was partially dependent on anti-oxidative process. Knockdown of TOM40 by adeno-associated virus (AAV) in the hippocampus or amygdala dramatically reduced the effect of ZBD-2 on PPD, indicating that TOM40 mediates the effect of ZBD-2 on PPD. CONCLUSIONS: TOM40 is required for the effect of ZBD-2 on treating anxiety and depression in PPD mice. This study reveals the role of microglia TSPO in PPD development and provides the new therapeutic strategy for PPD.

Integrative Metabolomic and Transcriptomic Landscape during Akebia trifoliata Fruit Ripening and Cracking.

Akebia trifoliata fruit is prone to crack after ripening, but little is known about the mechanism underlying the cracking process. This study integrated transcriptomic and metabolomic data, revealing significant changes in 398 metabolites and 8414 genes during ripening and cracking, mainly impacting cell-wall metabolism. Multi-omics joint analysis indicated that genes related to polygalacturonase, pectate lyase, α-amylase, and glycogen phosphorylase were up-regulated after cracking, degrading cell wall and starch. Concurrently, diminished photosynthetic metabolism and heightened phenylpropanoid metabolism suggested alterations in cuticle structure, potentially impacting cell-wall robustness. Numerous auxin and abscisic acid signaling-related genes were expressed, and we assume that they contributed to the promoting peel growth. These alterations collectively might compromise peel strength and elevate expanding pressure, potentially leading to A. trifoliata cracking. Transcription factors, predominantly ethylene response factors and helix-loop-helix family members, appeared to regulate these metabolic shifts. These findings provide valuable insights into A. trifoliata cracking mechanisms; however, direct experimental validation of these assumptions is necessary to strengthen these conclusions and expedite their commercial utilization.

Latest Advances in Active Materials for Food Packaging and Their Application.

Food packaging plays a pivotal role in preserving the quality and safety of food products while extending their shelf life [...].

Effect of pasteurization processing and storage conditions on softening of acidified chili pepper: Pectin and it related enzymes.

The softening of acidified chili peppers induced by processing and storage has become a major challenge for the food industry. This study aims to explore the impact of pasteurization techniques, thermal processing (TP), high-pressure processing (HPP), addition of sodium metabisulfite (SMS), and storage conditions (25 °C, 37 °C, and 42 °C for 30 days) on the texture-related properties of acidified chili pepper. The results showed that the textural properties of samples were destructed by TP (the hardness of samples decreased by 19.43 %) but were less affected by HPP and SMS. Compared with processing, storage temperature had a more dominant impact on texture and pectin characteristics. With increased storage temperature, water-solubilized pectin fraction content increased (increased by 160.99 %, 136.74 %, and 13.01 % in TP, HPP, and SMS-stored groups, respectively), but sodium carbonate-solubilized pectin fraction content decreased (decreased by 29.84 %, 26.81 %, and 8.60 % in TP-, HPP-, and SMS-stored groups, respectively), especially in TP-stored groups. Multivariate data analysis showed that softening was more closely related to pectin conversion induced by acid hydrolysis and pectinase depolymerization. This finding offers new perspectives for the production of acidified chili pepper.

PJA1 mediates the effects of astrocytic GPR30 on learning and memory in female mice.

Hormone replacement therapy (HRT) is not recommended for treating learning and memory decline in menopausal women because it exerts adverse effects by activating classic estrogen receptors ERα and ERß. The membrane estrogen receptor G protein-coupled receptor 30 (GPR30) has been reported to be involved in memory modulation; however, the underlying mechanisms are poorly understood. Here, we found that GPR30 deletion in astrocytes, but not in neurons, impaired learning and memory in female mice. Astrocytic GPR30 depletion induced A1 phenotype transition, impairing neuronal function. Further exploration revealed that Praja1 (PJA1), a RING ubiquitin ligase, mediated the effects of astrocytic GPR30 on learning and memory by binding to Serpina3n, which is a molecular marker of neuroinflammation in astrocytes. GPR30 positively modulated PJA1 expression through the CREB signaling pathway in cultured murine and human astrocytes. Additionally, the mRNA levels of GPR30 and PJA1 were reduced in exosomes isolated from postmenopausal women while Serpina3n levels were increased in the plasma. Together, our findings suggest a key role for astrocytic GPR30 in the learning and memory abilities of female mice and identify GPR30/PJA1/Serpina3n as potential therapeutic targets for learning and memory loss in peri- and postmenopausal women.

Chemometric Classification of Apple Cultivars Based on Physicochemical Properties: Raw Material Selection for Processing Applications.

Apple cultivars exhibit significant diversity in fruit quality traits, creating distinct consumption scenarios. This study aimed to assess the physicochemical parameters and sensory attributes differences among fifteen apple cultivars and identify characteristic qualities suitable for various processed apple products using chemometric analysis. Relatively large differences were registered between cultivars for deflection, peel color, titratable acidity (TA), the ratio of total soluble solid to titratable acidity (TSS/TA), hardness, soluble sugar, and volatile organic compound contents. Sensory results showed significant differences existed among the preferences for different processed products. Based on the above results, all cultivars could be distinguished into three main clusters. Cluster I (i.e., Aziteke, Bakeai, Magic Flute, Royal Gala, Red General, Red Delicious, and Zhongqiuwang) demonstrated favorable appearance, high sensory scores, and rich aroma volatile compounds, making them suitable for direct consumption. Cluster II (i.e., Fuburuisi, Sinike, Honglu, and Huashuo) exhibited a higher sugar and acid content, making them suitable for apple juice production. Cluster III (i.e., Miqila, Honey Crisp, Shandong Fuji, and Yanfu 3) were more suitable for fresh-cut apples due to their good flavor and undesirable appearance. Several chemometric analyses effectively assessed differences among apple cultivars.

A Novel Strategy to Improve Cloud Stability of Orange-Based Juice: Combination of Natural Pectin Methylesterase Inhibitor and High-Pressure Processing.

This study investigated the prospect of producing cloud-stable orange-based juice by combining high-pressure processing (HPP) with a natural kiwifruit pectin methylesterase inhibitor (PMEI) during chilled storage. Kiwifruit is rich in a PMEI, which greatly improves the cloud loss caused by the pectin methylesterase (PME) demethylation of pectin. The results show that the cloud loss of orange juice occurred after 3 days, while the orange-kiwifruit mixed juice and kiwifruit puree were cloud stable during 28 days' storage. Although, the kiwifruit puree contained larger particles compared to the orange juice, its higher viscosity and solid-like behavior were dominant, improving the cloud stability of the juice systems. In addition, the particle size distribution and rheological properties were highly related to PME activity, PMEI activity, and pectin characterization. The kiwifruit PMEI showed higher resistance to HPP and storage time than PME. More water-solubilized pectin fractions with a high molecular mass were found in the kiwifruit puree, leading to its high viscosity and large particle size, but a more chelator-solubilized pectin fraction with a low esterification degree was observed in the orange juice, resulting in its cloud loss. In general, the outcome of this work provides a novel strategy to improve the cloud stability of orange-based juices using natural PMEIs and nonthermal processing technologies.

Preparation of garlic stem cellulose nanocrystal/leaf extract/chitosan film for black garlic preservation by electrostatic spraying.

In this study, a novel active chitosan (CH) packaging film that incorporates garlic leaf extract (GL) and stem cellulose nanocrystals (CNC) was prepared. The addition of CNC to the CH film increased its tensile strength, hydrophilicity, thermal stability, and water/oxygen barrier and decreased its water contact angle and weight-loss rate, while the addition of GL greatly enhanced its antioxidant and antibacterial activities. SEM and AFM analyses showed that the CNC agglomerates and deposits in the lower layer and the surface roughness of the film was the highest at 1.2 % concentration. The optimal composition of the film was determined to be 0.8 % CNC and 4 % GL by the fuzzy mathematics evaluation method. Then, black garlic was preserved with the optimized coating by electrostatic spraying and was found to slow water loss and migration, while its excellent antioxidant activities decreased the degree of browning during 90 d of storage.

Effect of electroacupuncture combined with rehabilitation training on hypertension: A systematic review and meta-analysis.

To evaluate the clinical effects of electroacupuncture combined with rehabilitation training (EART) versus conventional rehabilitation training (CRT) on hypertension. Multiple databases like PubMed, Embase, Web of Science and Wanfang database, China National Knowledge Internet database, and Chinese Biological Medical database were used to search for the relevant studies and full-text articles involved in evaluating EART versus CRT with hypertension. Review Manager 5.4 was used to estimate the effects of the results among included articles. Forest plots, sensitivity analysis, and funnel plots were also conducted on the included articles. In this meta-analysis study, there were 9 relevant studies were eventually satisfied the included criteria. There were significant differences between EART group and CRT group in systolic blood pressure after treatment (MD -16.62, 95 %CI = -21.84 to -11.39; P < 0.00001), diastolic blood pressure after treatment (MD = -16.03, 95 % CI = -21.55 to -10.50; P < 0.00001), and effective rate (MD = 1.22, 95 % CI = 1.13 to 1.32; P < 0.00001). Sensitivity analysis and funnel chart demonstrated that the study was robust and limited publication bias was observed. Our data showed that EART was clinically more significant than CRT in hypertension. Further studies need to be performed using large relevance references to verify the effectiveness of EART in the treatment of hypertension.

Evaluation of Radio Frequency-Assisted Enzymatic Extraction of Non-Anthocyanin Polyphenols from Akebia trifoliata Flowers and Their Biological Activities Using UPLC-PDA-TOF-ESI-MS and Chemometrics.

A new radio frequency heating-assisted enzymatic extraction (RF-E) method is applied for the determination of phenolic compounds in Akebia trifoliata flowers, compared with hot water, acidified ethanol (EtOH), and enzymatic-assisted (EA) extractions. Non-anthocyanin polyphenol profiles, antibacterial, angiotensin-converting enzyme (ACE) inhibitory, anti-inflammatory activities, and structures of extracts are evaluated. Results show no significant differences in the extraction of total flavonoid content (15.85-16.63 mg QEs/g) and ACE inhibitory activity (51.30-52.86%) between RF-E and EA extracts. RF-E extract shows the highest anti-inflammatory activities. FTIR and UV spectra reveal that acidified EtOH treatment has a significant effect on the structure of the extract due to its highest flavonoid content (20.33 mg QEs/g), thus it has the highest antibacterial activity against Staphylococcus aureus and Escherichia coli. Sixteen non-anthocyanin polyphenols are identified by UPLC-PDA-TOF-ESI-MS and RF pre-treatment did not cause significant compound degradation. The chemometric analysis shows that enzymatic hydrolysis significantly increased biological activities, and the presence of non-anthocyanin polyphenols correlates well with ACE inhibitory and anti-inflammatory activities. Accordingly, A trifoliata flowers have potential as reagents for the food and pharmaceutical industries due to their abundant polyphenols that could be extracted efficiently using RF-E.

Effects of pasteurization technologies and storage conditions on the flavor changes in acidified chili pepper.

In this study, the effects of thermal processing (TP), high pressure processing (HHP), and preservatives addition, i.e. sodium metabisulfite (SMS), on flavor changes in acidified chili peppers were compared. In addition, their changes during different storage periods (25, 37, and 42 °C for 30 days) were also investigated. The results indicate that TP clearly changed the flavor properties of acidified chili peppers compared to other processing, such as an increase in organic acid contents and titratable acid (TA) values but a decrease in pH value, free amino acid (FAA) concentrations, and some aromatic compound contents (e.g., esters and aldehydes). For SMS groups, more biter FAAs and higher alcohol concentrations were detected. Some terpenes (e.g., ß-ocimene) significantly increased in samples after HPP (P < 0.05). In addition, storage conditions also clearly affected their flavor, particularly for high storage temperature. During storage, pH fast decreased but TA values and organic acids increased; FAAs firstly increased but followed decreased; esters and terpenes were the main compounds decreasing. Furthermore, some off-flavor related compounds were produced when samples were stored at high temperature, such as furans, aldehydes, and oxides. The outcome of this study could provide new insights into the effects of processing and storage conditions on flavor changes and guide production for the acidified chili pepper industry.

Effects of antiepileptic drugs polytherapy on pregnancy outcomes in women with epilepsy: An observation study in northwest China.

OBJECTIVE: The management of pregnant women with epilepsy (WWE) treated with antiepileptic drugs (AEDs) polytherapy poses a great challenge. The purpose of this study was to evaluate the major congenital malformations (MCMs) associated with AED polytherapy, to assess the impacts of polytherapy regimens on seizure control and breastfeeding, and to determine the potential predictors for pregnancy outcomes. METHODS: This study was based on prospectively acquired data from a registry enrolling WWE in early pregnancy from Feb 2010 to July 2019, in which 123 pregnancies in 110 WWE were exposed to 27 different AED combinations. RESULTS: There were 123 pregnancies in 110 WWE analyzed in our study. The live birth rate was 86.2 % and the risk of MCMs was 10.4 %. Multivariate analysis indicated that prenatal exposure to phenobarbital (odds ratio [OR], 17.424; 95 %CI, 1.510-201.067; P = 0.022) and topiramate (OR, 9.469; 95 %CI, 1.149-62.402; P = 0.036) was associated with increased risk of MCMs. Valproate (OR, 4.441; 95 %CI, 1.165-16.934; P = 0.029), phenobarbital (OR, 13.636; 95 %CI, 2.146-86.660; P = 0.006) and topiramate (OR, 7.527; 95 %CI, 1.764-32.118; P = 0.006) were significantly correlated with adverse pregnancy outcomes. Among 67 pregnancies in four combinations over 10 patients, 15 (22.4 %) remained seizure free through pregnancy, seizure frequency increased in 17 (25.4 %), decreased in 24 (35.8 %) women, in 26 (38.8 %) remained unchanged. Only 23.6 % of mothers undertook exclusive breastfeeding. Planned pregnancy was the only independent factor significantly associated with decreased risk of adverse pregnancy outcomes (OR, 0.139; 95 % CI, 0.051-0.382; P < 0.001). Notably, no adverse pregnancy outcome was recorded in pregnancies exposed to the combination of lamotrigine plus levetiracetam. CONCLUSION: Prenatal exposure to the combinations containing valproate, phenobarbital, or topiramate was associated with increased risk of adverse pregnant outcomes. AED-related teratogenicity may be reduced by planned pregnancy in WWE exposed to polytherapy. Our findings also suggest the combination of lamotrigine and levetiracetam seems to be most desirable to balance seizure control and fetal safety.

Effect of radio frequency, ultrasound, microwave-assisted papain, and alcalase hydrolysis on the structure, antioxidant activity, and peptidomic profile of Rosa roxburghii Tratt. seed protein.

Traditional enzymatic hydrolysis methods have defects such as low efficiency and poor bioactivity in the production of active peptides. In this study, radio frequency (RF) technology was innovatively used to assist the hydrolysis of Rosa roxburghii Tratt. seed protein (RTSP) by papain and alcalase. RF-assisted hydrolysis was compared with ultrasound-(US) and microwave (MW)-assisted techniques in terms of the degree of hydrolysis, structure, antioxidant properties, and changes in the peptidome of the hydrolysates to clarify the mechanism of functional change of physically-assisted hydrolysate. All three methods improved hydrolysis efficiency. The degree of hydrolysis (DH) of papain group increased from 6.38% to 7.97%, 9.97% and 8.37% after US-, MW- and RF-assisted hydrolysis, respectively, while the DH of alcalase-treated group increased from 21.13% to 25.66%, 26.03%, and 23.01%, respectively. The in vitro antioxidant capacity and intracellular antioxidant capacity of RTSP and its hydrolysates were measured and evaluated by fuzzy statistical evaluation, and MW-assisted alcalase hydrolysis had the highest in vitro and intracellular antioxidant activity scores of 0.713 and 0.820, respectively. Fourier transform infrared and amino acid composition analysis explained the enhanced antioxidant properties of the hydrolysates. Further peptide profiling showed the physical assistance led to an increase in the species and contents of small molecule antioxidant peptides compared to enzyme treatment alone. Pearson's linear correlation analysis showed that AY, LY, IY, PHW, SVL, LHL, YYV, VYY, and NHAV were significantly correlated with the antioxidant properties of hydrolysates. Our data suggested that physical assistance such as US, MW, and RF were effective to improve the efficiency of enzymatic hydrolysis and produce novel antioxidant peptides. PRACTICAL APPLICATION: In this study, it was found that electromagnetic wave-assisted enzymatic hydrolysis could improve the efficiency of hydrolysis and enhance the antioxidant activity of hydrolysates compared to unassisted means. Compared with MW treatment, RF has the comparable hydrolysis effect, but has the advantages of high penetration ability, good uniformity, and low energy consumption and has greater potential for the production of bioactive peptides.

Tanshinone IIA improves contextual fear- and anxiety-like behaviors in mice via the CREB/BDNF/TrkB signaling pathway.

Posttraumatic stress disorder (PTSD) is one of the most common psychiatric diseases, which is characterized by the typical symptoms such as re-experience, avoidance, and hyperarousal. However, there are few drugs for PTSD treatment. In this study, conditioned fear and single-prolonged stress were employed to establish PTSD mouse model, and we investigated the effects of Tanshinone IIA (TanIIA), a natural product isolated from traditional Chinese herbal Salvia miltiorrhiza, as well as the underlying mechanisms in mice. The results showed that the double stress exposure induced obvious PTSD-like symptoms, and TanIIA administration significantly decreased freezing time in contextual fear test and relieved anxiety-like behavior in open field and elevated plus maze tests. Moreover, TanIIA increased the spine density and upregulated synaptic plasticity-related proteins as well as activated CREB/BDNF/TrkB signaling pathway in the hippocampus. Blockage of CREB remarkably abolished the effects of TanIIA in PTSD model mice and reversed the upregulations of p-CREB, BDNF, TrkB, and synaptic plasticity-related protein induced by TanIIA. The molecular docking simulation indicated that TanIIA could interact with the CREB-binding protein. These findings indicate that TanIIA ameliorates PTSD-like behaviors in mice by activating the CREB/BDNF/TrkB pathway, which provides a basis for PTSD treatment.