Walnut protein isolate-epigallocatechin gallate nanoparticles: A functional carrier enhanced stability and antioxidant activity of lycopene.

Walnut isolate protein (WPI)-epigallocatechin gallate (EGCG) conjugates can be employed to creat food-grade delivery systems for preserving bioactive compounds. In this study, WPI-EGCG nanoparticles (WENPs) were developed for encapsulating lycopene (LYC) using the ultrasound-assisted method. The results indicated successful loading of LYC into these WENPs, forming the WENPs/LYC (cylinder with 200-300 nm in length and 14.81-30.05 nm in diameter). Encapsulating LYC in WENPs led to a notable decrease in release rate and improved stability in terms of thermal, ultraviolet (UV), and storage conditions compared to free LYC. Simultaneously, WENPs/LYC exhibited a synergistic and significantly higher antioxidant activity with an EC50 value of 23.98 µg/mL in HepG2 cells compared to free LYC's 31.54 µg/mL. Treatment with WENPs/LYC led to a dose-dependent restoration of intracellular antioxidant enzyme activities (SOD, CAT, and GSH-Px) and inhibition of intracellular malondialdehyde (MDA) formation. Furthermore, transcriptome analysis indicated that enrichment in glutathione metabolism and peroxisome processes following WENPs/LYC addition. Quantitative real-time reverse transcription PCR (qRT-PCR) verified the expression levels of related genes involved in the antioxidant resistance pathway of WENPs/LYC on AAPH-induced oxidative stress. This study offers novel perspectives into the antioxidant resistance pathway of WENPs/LYC, holding significant potential in food industry.

Novel bigels based on walnut oil oleogel and chitosan hydrogel: Preparation, characterization, and application as food spread.

This study developed new biphasic gel systems containing a walnut oil-based oleogel and a chitosan hydrogel and evaluated the application on food spread. The effects of different oleogelators [γ-oryzanol/ß-sitosterol (γ-ORY/ß-SIT), candelilla wax/span 65 (CW/SA), and mono- and diglycerides of fatty acids] were explored. Rheological analysis showed that γ-ORY/ß-SIT-based bigel had the strongest gel strength, but XRD confirmed that ß' crystal form (d = 3.72 Å, 4.12 Å) was predominantly in the CW/SA-based bigel, which was more appropriate for application as spread. The characteristics of CW/SA-based bigel with different oleogel fractions (40-80 wt%) were investigated. The microscopic images indicated that the hydrogels were dispersed as small droplets in the oleogels after oleogel fraction reaching 60 %. The highest crystallinity was achieved when the oleogel fraction was 60 %, and its oil binding capacity was 96.49 %. Textural analysis showed that the CW/SA-based bigel (OG-60 %) had similar properties with commercial spread B, and can be used as a partial replacement for spread B. Replacing 75 % of the commercial spread B with the bigel was found to be optimal and displayed acceptable sensory features. This study developed a healthy bigel based on walnut oil and provided the in-depth information for bigels as an alternative to plastic fats.

Does skin prick test response intensity predict symptom severity and efficacy of subcutaneous immunotherapy in allergic rhinitis?

OBJECTIVES: To investigate the effect of response intensity of allergen skin prick test (SPT) on symptom severity and long-term efficacy of dust mite subcutaneous immunotherapy (SCIT) in allergic rhinitis (AR). METHODS: AR Patients diagnosed with dust mite allergy and completed 3 years of SCIT were collected and classified into three groups: grade 2 (SPT of + +), grade 3 (SPT of + + +) and grade 4 (SPT of + + + +). Comparisons between groups were performed to examine the associations of SPT categories and symptom severity and the long-term efficacy of SCIT in AR. RESULTS: 181 AR patients were included. There was no significant difference in the baseline TNSS, SMS, RQLQ and VAS, and particularly to symptom severity grading among three SPT grade groups (P > 0.05). The moderate-severe AR was more likely to be smoking and accompany with asthma and had higher prevalence of sensitization to cockroach, mixed grass and tree pollen than mild AR (P < 0.05). Prevalence of sensitization to cockroach, mixed grass, ragweed and animal dander was increased in AR patients with asthma and allergic conjunctivitis (P < 0.05). Furthermore, after 3 years of SCIT, no statistical differences in TNSS, SMS, RQLQ, VAS and long-term efficacy were observed among the three SPT grade groups (P > 0.05). Similarly, long-term outcomes of patients with different SPT grades did not differ among different clinical characteristics and different efficacy determination criteria (P > 0.05). CONCLUSIONS: The SPT response intensity cannot be used as an objective evaluation index for symptom severity and the long-term efficacy of SCIT in AR patients.

Effects of Metabolic Syndrome and its components on the postoperative recurrence in Chronic Rhinosinusitis with Nasal Polyps' patients.

OBJECTIVES: Metabolic Syndrome (MetS) has been established as a significant factor in the pathogenesis of numerous chronic inflammatory conditions. However, its role in Chronic Rhinosinusitis with Nasal Polyps (CRSwNP) is unknown. This study aims to investigate the association between MetS, its components, and the risk of postoperative recurrence in Chinese patients with CRSwNP. METHODS: A retrospective cohort study was conducted on CRSwNP patients who underwent endoscopic sinus surgery in our hospital. Patients were divided into MetS and non-MetS groups, and the clinical characteristics and recurrence rates were compared. All CRSwNP patients were followed up for more than 2-years and further categorized into non-recurrent and recurrent groups. Binary logistic regression analyses were performed to examine the effects of MetS and its components on the risk of postoperative recurrence. RESULTS: A total of 555 CRSwNP patients were enrolled in the present study, 157 patients were included in the MetS group and 398 patients were categorized into the non-MetS group. The recurrence rate in the MetS group was significantly higher compared to the non-MetS group (p < 0.05). The rate of MetS, overweight or obesity, hyperglycemia and dyslipidemia were higher in the recurrent group in comparison with the non-recurrent group (p < 0.05). Multivariate logistic regression analysis suggested that MetS, overweight or obesity, hyperglycemia, dyslipidemia, and accompanying allergic rhinitis were associated with the risk of postoperative recurrence of CRSwNP (p < 0.05). Moreover, adjusted and unadjusted regression models showed that MetS was an independent risk factor for postoperative recurrence of CRSwNP, and the risk increased with more components of MetS included (p < 0.05). CONCLUSION: Our findings revealed that MetS independently increased the risk of postoperative recurrence in patients with CRSwNP, with the risk escalating as the number of MetS components increased. Moreover, accompanying allergic rhinitis was also demonstrated to be a potential risk factor for CRSwNP recurrence. LEVEL OF EVIDENCE: Level 4.

Screening and Constructing of Novel Angiotensin I-Converting Enzyme Inhibiting Peptides from Walnut Protein Isolate and Their Mechanisms of Action: A Merged In Silico and In Vitro Study.

Angiotensin I-converting enzyme (ACE)-inhibiting peptides were isolated from walnut protein isolate (WPI) using ultrasound-assisted extraction. This study aimed to assess the impact of ultrasonic pretreatment on the physicochemical properties of WPI. The optimal extraction conditions for WPI were determined as a 15-min ultrasonic treatment at 400 W. Subsequently, the hydrolysate exhibiting the highest in vitro ACE-inhibiting activity underwent further processing and separation steps, including ultrafiltration, ion exchange chromatography, liquid chromatography-tandem mass spectrometry, ADMET screening, and molecular docking. As a result of this comprehensive process, two previously unidentified ACE-inhibiting peptides, namely Tyr-Ile-Gln (YIQ) and Ile-Tyr-Gln (IYQ), were identified. In addition, a novel peptide, Ile-Lys-Gln (IKQ), was synthesized, demonstrating superior ACE-inhibiting activity and temperature stability. In silico analysis estimated an in vivo utilization rate of 21.7% for IKQ. These peptides were observed to inhibit ACE through an anti-competitive mechanism, with molecular docking simulations suggesting an interaction mechanism involving hydrogen bonding. Notably, both IYQ and IKQ peptides exhibited no discernible toxicity to HUVECs cells and promoted nitric oxide (NO) generation. These findings underscore the potential of ultrasonicated WPI in the separation of ACE-inhibiting peptides and their utility in the development of novel ACE inhibitors for functional food applications.

Effects of Metabolic Syndrome and its components on the postoperative recurrence in Chronic Rhinosinusitis with Nasal Polyps' patients

Abstract Objectives Metabolic Syndrome (MetS) has been established as a significant factor in the pathogenesis of numerous chronic inflammatory conditions. However, its role in Chronic Rhinosinusitis with Nasal Polyps (CRSwNP) is unknown. This study aims to investigate the association between MetS, its components, and the risk of postoperative recurrence in Chinese patients with CRSwNP. Methods A retrospective cohort study was conducted on CRSwNP patients who underwent endoscopic sinus surgery in our hospital. Patients were divided into MetS and non-MetS groups, and the clinical characteristics and recurrence rates were compared. All CRSwNP patients were followed up for more than 2-years and further categorized into non-recurrent and recurrent groups. Binary logistic regression analyses were performed to examine the effects of MetS and its components on the risk of postoperative recurrence. Results A total of 555 CRSwNP patients were enrolled in the present study, 157 patients were included in the MetS group and 398 patients were categorized into the non-MetS group. The recurrence rate in the MetS group was significantly higher compared to the non-MetS group (p< 0.05). The rate of MetS, overweight or obesity, hyperglycemia and dyslipidemia were higher in the recurrent group in comparison with the non-recurrent group (p< 0.05). Multivariate logistic regression analysis suggested that MetS, overweight or obesity, hyperglycemia, dyslipidemia, and accompanying allergic rhinitis were associated with the risk of postoperative recurrence of CRSwNP (p< 0.05). Moreover, adjusted and unadjusted regression models showed that MetS was an independent risk factor for postoperative recurrence of CRSwNP, and the risk increased with more components of MetS included (p< 0.05). Conclusion Our findings revealed that MetS independently increased the risk of postoperative recurrence in patients with CRSwNP, with the risk escalating as the number of MetS components increased. Moreover, accompanying allergic rhinitis was also demonstrated to be a potential risk factor for CRSwNP recurrence. Level of evidence: Level 4.

Astaxanthin protects against pirarubicin-induced H9c2 cardiomyocytes by adjusting microRNA-494-3p-mediated MDM4/p53 signalling pathway.

PURPOSE: Pirarubicin (THP) is an antitumour drug widely used in clinical practice, but its cardiotoxicity limits its application. THP cardiotoxicity must be treated as soon as possible. There is an urgent need to find drugs that alleviate THP cardiotoxicity. The purpose of this study was to investigate the effects and mechanisms of Astaxanthin (AST) on THP-induced cardiomyocytes. METHODS: Rat cardiomyocytes H9c2 were induced with THP. The effects of AST on THP-induced H9c2 and its mechanism were investigated by CCK8, reactive oxygen species assay, tunnel assay, flow cytometry, RT-qPCR, and Western blot. RESULTS: AST increased cell viability, inhibited apoptosis and accelerated cell cycle progression, reduced oxidative damage and inflammatory response in THP-induced H9c2; down-regulated miR-494-3p expression, promoted MDM4 expression, inhibited p53 activation, and suppressed apoptosis-related protein expression. Overexpression of MiR-494-3p reversed the above effects of AST. CONCLUSIONS: AST can inhibit H9c2 apoptosis induced by THP and attenuate H9c2 damage by THP, which may be achieved by downregulating miR-494-3p, upregulating MDM4, and inhibiting p53.

Characterization of walnut protein isolate-polyphenol nanoconjugates for the developing a food-grade delivery system.

This study investigated the effects of the interaction of walnut protein isolate (WPI) with epigallocatechin gallate (EGCG), chlorogenic acid (CLA), (+)-catechin (CA), and ellagic acid (EA) on the structural and functional properties of proteins. The results for polyphenol binding equivalents and content of free amino and sulfhydryl groups as well as those from sodium dodecyl sulfate‒polyacrylamide gel electrophoresis confirmed the covalent interaction between WPI and the polyphenols. The binding capacities of the WPI-polyphenol mixtures and conjugates were as follows: WPI-EGCG > WPI-CLA > WPI-CA > WPI-EA. Fourier transform infrared spectroscopy (FTIR) and fluorescence spectrum analysis identified changes in the protein structure. The conjugation process obviously increased the polyphenols' antioxidant properties and the surface hydrophobicity was substantially reduced. WPI-EGCG conjugates had the best functional properties, followed by WPI-CLA, WPI-CA, and WPI-EA. Lycopene (LYC) was loaded into nanocarriers by WPI-EGCG self-assembly. These results indicated that WPI-polyphenol conjugates can be utilized to develop food-grade delivery systems to protect chemically lipophilic bioactive compounds. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-023-05768-2.

MiR-494-3p aggravates pirarubicin-induced cardiomyocyte injury by regulating MDM4/p53 signaling pathway.

OBJECTIVE: Pirarubicin (THP) is a widely used antitumor drug in clinical practice, but its cardiotoxicity limits its use. There is an urgent need to find drugs to alleviate the cardiotoxicity of THP. This study aimed to investigate the effect and mechanism of miR-494-3p on THP-induced cardiomyocytes. METHODS: THP induced immortalized mouse cardiomyocytes HL-1, silenced or overexpressed miR-494-3p. The effects of miR-494-3p on HL-1 contained in THP were investigated by CCK8, flow cytometry, ROS detection, JC-1 mitochondrial membrane potential detection, TUNEL cell apoptosis detection, RT-qPCR, and Western blot. RESULTS: miR-494-3p could reduce cell viability, increase oxidative damage, and promote cell apoptosis; at the same time, it inhibited the expression of MDM4, promoted the activation of p53, and promoted the expression of apoptosis-related proteins. MiR-494-3p inhibitors have the opposite effect. CONCLUSION: miR-494-3p can aggravate THP damage to HL-1, which may be achieved by downregulating MDM4 and promoting p53. miR-494-3p is one of the important miRNAs in THP-induced cardiotoxicity, which provides theoretical support for its possible use as a therapeutic target for THP-induced cardiovascular disease.

Chitosan-pullulan films enriched with Artemisia annua essential oil: Characterization and application in grape preservation.

Composite films were prepared using a flow casting method, with chitosan and pullulan as film-forming agents and Artemisia annua essential oil as the UV absorber. The utility of the composite films for preserving grape berries was assessed. The effect of the added Artemisia annua essential oil on the physicochemical properties of the composite film was investigated to determine the optimal amount of essential oil that should be added to the composite film. When the Artemisia annua essential oil content was 0.8 %, the elongation at break of the composite film increased to 71.25 ± 2.87 % and the water vapor transmission rate decreased to 0.378 ± 0.007 g‧mm/(m2‧h‧kpa). The transmittance of the composite film was almost 0 % in the UV region (200-280 nm) and <30 % in the visible light region (380-800 nm), reflecting the UV absorption by the composite film. Additionally, the composite film extended the storage time of the grape berries. Therefore, the composite film containing Artemisia annua essential oil may be a promising fruit packaging material.

Structural and textural properties of walnut protein gels induced by ultrasound and transglutaminase: encapsulation and release of tea polyphenols.

This study investigated the synergy of ultrasonic and transglutaminase (TGase) treatment on the structural, physicochemical, rheological, gelation properties and controlled release properties of dehulled walnut proteins (DWP). The results showed that after ultrasonic-TGase treatment, the surface hydrophobicity was decreased, indicating the involvement of disulfide bonds in gel formation. Scanning electron microscopy (SEM) showed that ultrasonic-TGase treatment resulted in a more uniform and denser microstructure of DWP gels. Ultrasonic-TGase treatment changed the secondary structure of the DWP gels as determined by Fourier transform infrared spectroscopy, with an increase in α-helix, ß-turn and random coils and a decrease in ß-sheets. In addition, in vitro drug release profiles showed that ultrasonic-TGase treatment promoted the cross-linking of protein molecules and formed a dense network to embed tea polyphenols (TP), thereby slowing down the digestion of TP in simulated gastric fluid and achieving the purpose of slow-release in simulated intestinal fluid. Thus, the synergy of ultrasonic and TGase treatment might be an effective method to improve gel properties and expand the application of protein gels in the food industries. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-023-05756-6.

Chitosan-coated nanoliposomes for the enhanced stability of walnut angiotensin-converting enzyme (ACE) inhibitory peptide.

This study encapsulated walnut angiotensin-converting enzyme (ACE) inhibitory peptides within nanoliposomes and then modified them with chitosan. The resulting effect of the nanoliposome loading and chitosan coating on physicochemical characteristics, stability, bioactivity, chemical structure, and morphology of the encapsulated peptides was assessed. The resulting particle size and polymer dispersity index revealed that the chitosan-coated nanoliposomes loaded with walnut ACE inhibitory peptides (WAIP) (CL-P) exhibited higher physical stability compared with the nanoliposomes loaded with WAIP (L-P). The encapsulation efficiency (EE) of CL-P increased from 73.32% to 76.13% after chitosan modification, and the EE of L-P and CL-P could be maintained by storage at 4°C. In addition, the antioxidant activity and ACE inhibitory activity of the peptides were effectively protected by L-P and CL-P during storage. Fourier transform infrared spectroscopy showed that the nanoliposomes were bound in ionic form with both the peptides and chitosan. Transmission electron micrographs indicated the presence of vesicle-like carriers with a reservoir-type structure. This study highlights the potential of nanoliposomes and their modification with chitosan to increase the stability and bioactivity retention of ACE inhibitory peptides. PRACTICAL APPLICATION: Chitosan-coated nanoliposomes loaded with walnut ACE inhibitory peptides were prepared in this study. Chitosan coating increased nanoliposomes' encapsulation efficiency and provided higher physical stability. In addition, the bioactivity of the walnut ACE peptides was effectively protected during storage. This study was relevant for improving the storage and transportation used for nanoliposome systems applied in the food and health product industry.

Tumor necrosis factor α-induced protein 3 mediates inflammation and neuronal autophagy in Parkinson's disease via the NFκB and mTOR pathways.

This study aimed to probe the function of tumor necrosis factor α-induced protein 3 (TNFAIP3) in the pathogenesis of Parkinson disease (PD) with its association with autophagy and inflammatory response. TNFAIP3 was reduced in the SN of PD patients (the GSE54282 dataset) and mice and in the MPP+-treated SK-N-SH cells. TNFAIP3 inhibited inflammatory response and enhanced autophagy, thereby alleviating PD in mice. NFκB and mTOR pathways were activated in the SN of PD mice and MPP+-treated cells. TNFAIP3 blocked the two pathways by preventing the p65 nuclear translocation and stabilizing DEPTOR, an endogenous inhibitor of mTOR. NFκB activator LPS and mTOR activator MHY1485 reversed the effects of TNFAIP3 on mitigation of injury in PD mice and in SK-N-SH cells induced with MPP+. Altogether, TNFAIP3 played a neuroprotective role in MPTP-induced mice by restricting NFκB and mTOR pathways.

Significance of leukocyte-specific transcript 1 levels in nasal mucosal tissue to predict recurrence of nasal polyps

Abstract Objective: Chronic Rhinosinusitis with Polyps (CRSwNP) is characterized by high heterogeneity and postoperative recurrence rate. This study aims to explore the clinical significance of tissue Leukocyte-Specific Transcript 1 (LST1) in predicting CRSwNP recurrence. Methods: We enrolled 62 CRSwNP patients including 30 primary CRSwNP and 32 recurrent CRSwNP patients, and 40 Healthy Controls (HC). Tissue samples were collected. Tissue LST1 expression was assessed by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), Western Blotting (WB) and Immunofluorescence (IF) staining. The predictive values of LST1 expression for CRSwNP postoperative recurrence were assessed through the Receiver Operating Characteristic (ROC) curves. Results: The tissue levels of LST1 were significantly increased in the CRSwNP group than the HC group, especially in the recurrent group, and the elevated LST1 mRNA levels were positively correlated with the peripheral eosinophil percentages, tissue eosinophil counts and percentages. IF staining results showed that the LST1 protein levels were higher in CRSwNP patients, especially in the recurrent patients than in the HC group. ROC curves highlighted that tissue LST1 levels were associated with recurrent CRSwNP and exhibited a higher predictive ability for postoperative CRSwNP recurrence. Conclusion: This was the first report suggesting that LST1 expression was upregulated and associated with mucosal eosinophil infiltration and CRSwNP recurrence. Tissue LST1 could be a promising biomarker for predicting postoperative recurrence in CRwNP patients.

Elevated body mass index increased the risk of recurrence in Chinese patients with chronic rhinosinusitis.

BACKGROUND: Elevated body mass index (BMI) has been associated with an increased prevalence of chronic rhinosinusitis (CRS). However, the impact of BMI on the risk of recurrence of CRS is unknown. This study aimed to investigate the association between BMI and the risk of CRS recurrence. MATERIAL AND METHODS: A retrospective cohort study was conducted and recruited 1057 CRS patients who underwent functional endoscopic sinus surgery (FESS). All subjects were classified into four groups: "underweight", "normal weight", "overweight", and "obese". Multivariate regression analysis was performed to examine the associations between BMI categories and other factors and the risk of CRS recurrence. RESULTS: The rate of recurrent CRS was significantly higher in the overweight group and obese group than in the normal weight group (P < 0.05). Unadjusted and adjusted logistic regression analyses demonstrated that overweight and obesity exhibited an increased risk of CRS recurrence as compared to patients with normal weight (P < 0.05). Furthermore, all patients were divided into primary CRS group and recurrent CRS group, and the BMI, duration of disease and rate of allergic rhinitis were vastly increased in the recurrent group than in the primary group (P < 0.05). Univariate and multivariate regression analysis showed that BMI and duration of disease were the dominant risk factors of CRS recurrence (P < 0.05). CONCLUSION: Overweight and obesity presented significant impacts on the CRS recurrence, and elevated BMI were associated with an increased risk of CRS recurrence independently from traditional risk factors. A longer duration of disease was correlated with a higher risk of CRS recurrence.

Significance of leukocyte-specific transcript 1 levels in nasal mucosal tissue to predict recurrence of nasal polyps.

OBJECTIVE: Chronic Rhinosinusitis with Polyps (CRSwNP) is characterized by high heterogeneity and postoperative recurrence rate. This study aims to explore the clinical significance of tissue Leukocyte-Specific Transcript 1 (LST1) in predicting CRSwNP recurrence. METHODS: We enrolled 62 CRSwNP patients including 30 primary CRSwNP and 32 recurrent CRSwNP patients, and 40 Healthy Controls (HC). Tissue samples were collected. Tissue LST1 expression was assessed by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), Western Blotting (WB) and Immunofluorescence (IF) staining. The predictive values of LST1 expression for CRSwNP postoperative recurrence were assessed through the Receiver Operating Characteristic (ROC) curves. RESULTS: The tissue levels of LST1 were significantly increased in the CRSwNP group than the HC group, especially in the recurrent group, and the elevated LST1 mRNA levels were positively correlated with the peripheral eosinophil percentages, tissue eosinophil counts and percentages. IF staining results showed that the LST1 protein levels were higher in CRSwNP patients, especially in the recurrent patients than in the HC group. ROC curves highlighted that tissue LST1 levels were associated with recurrent CRSwNP and exhibited a higher predictive ability for postoperative CRSwNP recurrence. CONCLUSION: This was the first report suggesting that LST1 expression was upregulated and associated with mucosal eosinophil infiltration and CRSwNP recurrence. Tissue LST1 could be a promising biomarker for predicting postoperative recurrence in CRwNP patients. LEVEL OF EVIDENCE: Level 5.

Effects of polysaccharide thickening agent on the preparation of walnut oil oleogels based on methylcellulose: Characterization and delivery of curcumin.

Walnut oil-based oleogels were prepared by the emulsion-templated method using methylcellulose at different concentrations and viscosities as the oleogelators and polysaccharides (sodium alginate, xanthan gum and κ-carrageenan) as the thickening agents. The microscopic properties, rheological properties and oil binding capacity (OBC) of the oleogels were evaluated. The intermolecular and intramolecular hydrogen bonding of polysaccharide stabilized the network structure of the oleogel. The increasing methylcellulose concentration contributed to forming a more stable interfacial layer and providing oleogel with a compact structure. κ-Carrageenan resulted in a better OBC (97.37 %) and rheological properties of the methylcellulose-based oleogel. When served as a delivery system of curcumin, the highest encapsulation rate of curcumin (38.06 %) was achieved by the κ-carrageenan oleogel. The structure of oleogels slowed down the release rate of free fatty acids and curcumin during the early stage of in vitro digestion and the κ-carrageenan oleogel exhibited the highest inhibiting effect. This finding suggests that the polysaccharide-based walnut oil oleogels had a firmer structure and could be a promising approach to deliver curcumin.

Identification of prognostic genes signature and construction of ceRNA network in pirarubicin treatment of triple-negative breast cancer.

BACKGROUND: The altered long non-coding RNA (lncRNA), circular RNA (circRNA) and mRNA expression in triple-negative breast cancer (TNBC) after pirarubicin (THP) treatment can be a critical factor in the development of tumor. Here, we identify a set of lncRNA, circRNA, and mRNA that can reveal the molecular target and molecular mechanism of THP, and can be used to predict the prognostic characteristics of TNBC. METHODS: Affymetrix GeneChip sequencing was performed to determine whether lncRNA, circRNA, and mRNA were changed in MDA-MB-231 cells after THP treatment, and qRT-PCR was used to verify the accuracy of GeneChip results. Bioinformatics methods were used to analyze the differentially expressed (DE) lncRNA, circRNA and mRNA, and the co-expression network and ceRNA network were constructed. The STRING database, Kaplan-meier Mapper database, GEPIA database, and Tumor Immunity Estimation Resource were used to screen hub genes with clinical value and important significance. RESULTS: THP 5 µM could significantly inhibit proliferation, migration and invasion of MDA-MB-231 cells for 24 h. 1547 DE lncRNAs, 4992 DE circRNAs, and 5777 DE mRNAs were identified. The reliability of the GeneChip was verified by qRT-PCR. An mRNA-lncRNA/circRNA co-expression network was constructed based on the Pearson correlation coefficient. Finally, we established a new ceRNA network, including three circRNAs, five miRNAs, and three mRNAs. The mRNAs are associated with immune infiltration. The mRNAs and miRNAs are significantly associated with survival outcomes in TNBC. CONCLUSION: The results reveal the molecular target and mechanism of THP treatment of TNBC. These ceRNA network can be used as molecular targets for the treatment of TNBC patients and as molecular biomarkers to predict patient prognosis.

Icariin alleviates atherosclerosis by regulating the miR-205-5p/ERBB4/AKT signaling pathway.

PURPOSE: Atherosclerosis (AS) is a cardiovascular disease that has become a major threat to public health worldwide. This study aims to elucidate the effect and mechanism of icariin (ICA) in treating atherosclerosis. METHODS: ApoE-/- mouse AS modeling, ELISA, and hematoxylin-eosin staining were conducted to explore whether icariin has a therapeutic effect on AS. The microRNA (miRNA) chips for ICA treatment of ApoE-/- AS mice were developed; in silico analyses were performed, and signaling pathways were identified. Oxidized low-density lipoprotein (Ox-LDL) was used to induce human aortic vascular smooth muscle cells (HAVSMCs) to build an in vitro AS cell model. Moreover, miR-205-5p was silenced. Finally, cell viability was detected by MTT assay, cell apoptosis by flow cytometry and Western blot, and cell migration by the scratch test. RESULTS: ICA could reduce lipid accumulation in the blood vessels of mice and plaque formation to treat AS. ICA promoted apoptosis and inhibited cell migration of HAVSMCs induced by ox-LDL. Moreover, cell proliferation and migration were inhibited via ICA, which was restored by miR-205-5p silencing. CONCLUSION: ICA can alleviate AS and inhibit the proliferation and migration of HAVSMCs induced by ox-LDL, potentially mediated by the upregulation of miR-205-5p.

Novel angiotensin I-converting enzyme (ACE) inhibitory peptides from walnut protein isolate: Separation, identification and molecular docking study.

Walnut protein isolate was hydrolyzed using alcalase® to obtain angiotensin-I-converting enzyme (ACE) inhibitory (ACEI) peptides. The components with high ACEI activity were successfully purified from walnut protein isolate hydrolysates (WPIH) by ultrafiltration and G-25 gel chromatography. The 1520 peptides were identified by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Then the screening model of ACEI peptides was established by in silico approach. It was found that four ACEI active peptides (PPKP, YPQY, YLPP, and PKPP) were obtained with IC50 values ranging from 506 to 89 µmol/L, among which PPKP had the highest ACEI activity (IC50  = 89 ± 1 µmol/L). The four peptides mentioned above were novel, non-toxic, and resistant to gastrointestinal digestion. The molecular docking studies showed that the ACEI effect of ACEI peptide was mainly due to the interaction with residues of Gln281 and His353 in the ACE active pockets. In vivo availability of ACEI peptides showed that the probability of PPKP binding to ACE was 37.9% in the human body. Our studies suggest that the ACEI peptides derived from the WPIH can be considered functional foods that can prevent hypertension. PRACTICAL APPLICATIONS: Hypertension is a significant risk factor for cardiovascular and cerebrovascular disease, the leading cause of death worldwide. This study used a cost-effective method to isolate and identify potential ACEI peptides from the walnut meal. Since the walnut meal is often discarded in the processing of walnut products and thus pollutes the environment, the preparation of walnut meal into ACEI peptides can reduce the impact of hypertension on people and reduce environmental pollution. The experimental results show that walnut ACEI peptides are a safe and healthy nutritional product.