Propolis ethanol extract functionalized chitosan/Tenebrio molitor larvae protein film for sustainable active food packaging.

The application of novel insect proteins as future food resources in the food field has attracted more and more attention. In this study, a biodegradable antibacterial food packaging material with beneficial mechanical properties was developed using Tenebrio molitor larvae protein (TMP), chitosan (CS) and propolis ethanol extract (PEE) as raw materials. PEE was uniformly dispersed in the film matrix and the composite films showed excellent homogeneity and compatibility. There are strong intermolecular hydrogen bond interactions between CS, TMP, and PEE in the films, which exhibit the structure characteristics of amorphous materials. Compared with CS/TMP film, the addition of 3 % PEE significantly enhanced the elongation at break (34.23 %), water vapor barrier property (22.94 %), thermal stability (45.84 %), surface hydrophobicity (20.25 %), and biodegradability of the composite film. The composite film has strong antioxidant and antimicrobial properties, which were enhanced with the increase of PEE content. These biodegradable films offer an eco-friendly end-of-life option when buried in soil. Composite films can effectively delay the spoilage of strawberries and extend the shelf life of strawberries. Biodegradable active packaging film developed with insect protein and chitosan can be used as a substitute for petroleum-based packaging materials, and has broad application prospects in the field of fruits preservation.

Preparation of chitosan/Tenebrio molitor larvae protein/curcumin active packaging film and its application in blueberry preservation.

With growing concerns about postharvest spoilage of fruits, higher requirements have been placed on high-performance and sustainable active packaging materials. In this study, we prepared curcumin-based functional composite films using chitosan (CS) and Tenebrio molitor larvae protein (TMP) as the substrates. The effects of curcumin concentration on the structural and physicochemical properties of the composite films were determined. Curcumin was equally distributed in the polymer film through physical interactions. Furthermore, the curcumin composite film with 0.3 % addition exhibited a 27.39 % increase in elongation at break (EBA), a 37.04 % increase in the water vapor barrier, and strong UV-blocking properties and antioxidant activity compared with the control film (CS/TMP). The degradation experiment of the composite film on natural soil revealed that the composite film exhibited good biodegradability and environmental protection. Furthermore, the applicability of functional composite films for preserving blueberries was investigated. Compared with the control film and polyethylene (PE) films, the prepared composite films packaging treatment reduced the decay rate and weight loss rate of blueberries during storage, delayed softening and aging, and maintained the quality of blueberries. Using sustainable protein resources (TMP) and natural polysaccharides as packaging materials provides an economically, feasible and sustainable way to achieve the functional preservation of biomass materials.

Upgulation of lncRNA GASL1 inhibits atherosclerosis by regulating miR-106a/LKB1 axis.

BACKGROUND: Atherosclerosis (AS) is a common frequently-occurring disease in the clinic and a serious threat to human health. This research aimed to explore the value between GASL1 and AS. METHODS: The expression and values of GASL1 in AS patients were revealed by qRT-PCR and ROC curve. The HUVEC cells were induced by ox-LDL to construct in-vitro models. Cell viability was detected by MTT assay, and apoptosis was detected by flow cytometry. The inflammatory situation was reflected by the ELISA assay. Double luciferase reporter gene assay verified the regulatory relationship between GASL1 and miR-106a, miR-106a and LKB1. RESULTS: The levels of GASL1 was lower in AS group than those in control group. The value of GASL1 in predicting AS patients was also tested by the ROC curve. After HUVEC cells were induced by ox-LDL, the levels of GASL1 and LKB1 decreased significantly, while the level of miR-106a increased significantly. Upregulation of LKB1 reversed the effect of upregulation of GASL1 on viability, apoptosis, and inflammation of HUVEC cells induced by ox-LDL. CONCLUSION: Overexpression of GASL1 might suppress ox-LDL-induced HUVEC cell viability, apoptosis, and inflammation by regulating miR-106a/LKB1 axis.

Depleted miR-125a-5p Causes Vascular Endothelial Cell Dysfunction in Deep Vein Thrombosis by Targeting Angiopoietin 2.

Deep vein thrombosis (DVT) is a common and fatal disease with a pathology involving endothelial dysfunction. The present research aimed to address the potential clinical significance of miR-125a-5p in DVT and its effect on the dysfunction of Human umbilical vein endothelial cells (HUVECs). Serum miR-125a-5p levels were measured using RT-qPCR in 88 patients with DVT and 76 healthy controls. ROC was plotted to evaluate the diagnostic potential of miR-125a-5p. Spearman's correlation coefficient was performed to calculate the correlation between miR-125a-5p and clinical indicators. CCK-8, Transwell, and ELISA were employed to verify the effects of cell proliferation, migration, and inflammatory and adhesion molecules. Dual-luciferase reporter assay to analyze potential target for miR-125a-5p. Serum miR-125a-5p was reduced in patients with DVT compared with healthy controls (P < 0.001). ROC showed that miR-125a-5p significantly identified patients with DVT from the healthy controls (AUC = 0.834). Furthermore, serum miR-125a-5p was negatively correlated with inflammatory factors and coagulation factors. In in vitro studies, proliferation and migration of HUVECs were inhibited by suppressed miR-125a-5p, whereas inflammation and adhesion factors were considerably promoted (P < 0.05). Moreover, miR-125-5p directly targeted the 3'UTR of angiopoietin 2 (ANGPT2) and was negatively regulated. Finally, serum ANGPT2 was elevated in patients with DVT and was negatively correlated with serum miR-125a-5p. The current research demonstrated that decreased miR-125a-5p was a novel potential diagnostic biomarker for DVT and that it may be involved in DVT progression by targeting ANGPT2 to regulate endothelial dysfunction.

p53 Inhibition Provides a Pivotal Protective Effect against Cerebral Ischemia-Reperfusion Injury via the Wnt Signaling Pathway.

INTRODUCTION: Cerebral ischemia-reperfusion injury enhances brain injury and increases its morbidity and mortality. The purpose of our study was to further explore the specific pathogenesis of cerebral ischemia disease by studying the role of p53 in cerebral ischemia-reperfusion injury and its mechanism to provide a new target for the treatment of cerebral ischemia. METHODS: Middle cerebral artery occlusion (MCAo) was established in rats. The changes in p53 and apoptotic proteins in the rat model were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. The effects of p53 inhibitors on cerebral is-chemia-reperfusion injury in rats were evaluated by modified neurological severity score (mNSS) and infarct area. Subsequently, neural stem cells (NSCs) were isolated and cultured in vitro, and oxygen and glucose deprivation (OGD) was induced to establish an in vitro ischemia-reperfusion injury model. Cell viability and migration were detected by CCK-8 and transwell assays. Apoptosis of NSCs was detected by flow cytometry. Finally, protein expression in the Wnt pathway activated by p53 was detected by Western blotting. RESULTS: Compared with the sham group, p53 levels, mNSS, cerebral infarction area, and apoptosis were significantly increased in the MCAo group (p < 0.05). When the p53 inhibitor PFT-α was injected, the increase in these levels was reversed. Also, the viability and migration of cells decreased and apo-ptosis increased in the in vitro OGD model, whereas the viability, migration, and apoptosis were significantly reversed after the addition of p53 inhibitors (p < 0.05). Finally, p53 induced Wnt signaling pathway proteins ß-catenin and cyclin D1 decrease in the MCAo group, while p53 inhibitors reversed their inhibitory effect on the Wnt signaling pathway. CONCLUSION: We confirmed in vivo and in vitro that inhibition of p53 has a protective effect on the cerebral ischemia-reperfusion injury, which may be related to the activation of the Wnt signaling pathway.

[Inhibitory effect of Sirt3 on proliferation of vascular smooth muscle cells induced by angiotensin II].

OBJECTIVE: To detect the expression of Sirtuin3 (Sirt3) in mice vascular smooth muscle cells (VSMCs) and explore the effect of Sirt3 on VSMCs proliferation induced by angiotensin II (AngII). METHODS: The mRNA and protein expressions of Sirt3 in wild C57 mice VSMCs were assessed by RT-PCR and Western blotting, respectively. After the cells were exposed to various concentrations of AngII (10(-7);, 10(-6);, 10(-5); mol/L), the mRNA and protein expressions of Sirt3 were assessed again in the same way. The effect of Sirt3 on cell proliferation was observed with Edu (5-ethynyl -2'-deoxyuridine) kit after Sirt3 silencing by small interference RNA. RESULTS: Sirt3 was stably expressed in C57 mice VSMCs as revealed by Western blotting and RT-PCR, respectively. A significant increase was found in the mRNA and protein expressions of Sirt3 (P<0.01) after the stimulation of all the three different concentrations of AngII, particularly the 10(-6); mol/L group with the highest increase of Sirt3 expression, and there was a significant difference between the 10(-6); mol/L group and the other groups (P<0.05). The rate of cell proliferation in Sirt3 SiRNA group increased obviously compared to the control group (P<0.01), and the rate in the Sirt3 SiRNA+AngII group was also elevated significantly compared to the AngII group (P<0.01). CONCLUSION: The expression of Sirt3 is stable in VSMCs; AngII can elevate the mRNA and protein expression of Sirt3 and promote the cell proliferation, what's more, Sirt3 silencing would further increase the cell proliferation.

[Expression of SIRT1 in right auricle tissues and the relationship with oxidative stress in patients with atrial fibrillation].

AIM: To observe the expression of mammalian sirtuin 1 (SIRT1) in right auricle tissues in patients presenting with atrial fibrillation (AF) and make clear the relationship between SIRT1 expression and oxidative stress. METHODS: A total of 38 patients with rheumatic heart disease were divided into 2 groups: AF group (AF lasted more than 6 months, n=25) and SR (sinus rhythm) group (n=13). The expression of SIRT1 in right auricle tissues harvested during heart operations was detected by immunohistochemistry. Oxidative stress was estimated by the amounts of malondialdehyde (MDA) and metallothionein (MT) and the activity of superoxide dismutase (SOD) which were detected using thiobarbituric acid reaction (TBA), enzyme-linked immunosorbent assay (ELISA) and xanthine oxidase assay, respectively. RESULTS: Compared with the SR group, the expression of SIRT1 protein in the atrium significantly increased in AF group [P<0.05, (45.8±4.03)% vs (19.7±2.54)%]. In AF group, MDA was (7.24±1.05) nmol/mg, SOD (1034.25±84.32) U/mg and MT (7.21±1.46) µg/g, all being significantly higher than those in SR group[P<0.05, MDA: (3.01±0.47) nmol/mg; SOD: (723.63±65.23) U/mg; MT: (4.31±1.23) µg/g]. Spearman correlation indicated that the expression of SIRT1 had a significantly negative correlation with the amounts of MDA and MT and the activity of SOD (P<0.05, r=-0.447, -0.521, -0.394, respectively). CONCLUSION: The expression of SIRT1 increased in patients with AF. SIRT1 maybe effects the AF by means of inhibiting the process of oxidative stress.