Effects of ultrasonic treatment on the surface bacteria of Lyophyllum decastes during storage.

This study explores the relationship between the storage quality and bacterial microflora in the mushroom Lyophyllum decastes. The surface bacteria of L. decastes were separated by combining the traditional culture plate separation and 16S rRNA sequencing method, to study the effects of ultrasonic (US) treatment on the surface bacteria of L. decastes during storage. The results demonstrated that Pantoea agglomerans and Pseudomonas fluorescens were among the 15 culturable bacteria isolated with traditional plate method during storage, belonging to 2 phyla and 7 genera. US treatment could inhibit the growth and significantly increase cell membrane permeability, and contents extravasation in P. agglomerans, though its inhibitory effect on P. fluorescens was less. The 16S rRNA sequencing revealed, bacteria from 9 phyla and 35 genera were isolated, and P. fluorescens was the dominant species throughout the storage time. These results indicated that the composition of mushroom surface microflora of Control (CK) and US groups are similar, and the bacterial microflora networks analysis also showed a positive correlation. The KEGG annotation for the functional classification of the bacteria showed that a total of 328 pathways were acquired at the KEGG l3 level, and the relative abundance of membrane transport, amino acid metabolism, carbohydrate metabolism, and energy metabolism pathway was high. Moreover, the relative abundance of the surface bacteria of L. decastes also decreased. Hence, the US treatment had a better bacteriostatic effect, maintained the whiteness index and firmness, and improved the sensory quality of L. decastes during storage.

Ultrasonic treatment decreases Lyophyllum decastes fruiting body browning and affects energy metabolism.

Lyophyllum decastes is a common mushroom that is prone to browning during prolonged storage. In this study, the effects of ultrasonic treatment on metabolic gene expression, enzyme activity, and metabolic compounds related to L. decastes browning were investigated. Treatment of the fruiting body at 35 kHz and 300 W for 10 min reduced the browning index of L. decastes by 21.0 % and increased the L* value by 11.1 %. Ultrasonic treatment of the fruiting body resulted in higher levels of total phenols, flavonoids, and 9 kinds of amino acid with catalase (CAT) and peroxidase (POD) activities maintained at high levels. Higher cytochrome c oxidase (CCO), succinate dehydrogenase (SDH), phosphofructokinase (PFK), and pyruvate kinase (PK) activities may be ascribed to increased antioxidant capacity. Moreover, ultrasonication retained higher adenosine triphosphate (ATP) concentrations with an increased energy charge, while there were lower levels of adenosine diphosphate (ADP) and reduced and oxidized nicotinamide adenine dinucleotide (NADH and NAD+), respectively. Meanwhile, lower lignin contents were observed, along with retarded polyphenol oxidase (PPO) and lipoxygenase (LOX) activities. Lower PPO activity reduced the fruiting body enzymatic browning rate through decreased expression of LdPpo1, LdPpo2, and LdPpo3 during storage at 4 °C for 16 days. This activity may be used to determine the effectiveness of ultrasonication.

Therapeutic Effect of Catgut Implantation at Acupoint in a Mouse Model of Hepatocellular Carcinoma by Suppressing Immune Escape.

BACKGROUND: The occurrence and development of hepatocellular carcinoma (HCC) are closely related to immune function, as is the capacity of hepatoma cells to escape. Immunosurveillance is a key mechanism. Catgut implantation at acupoint (CIAA) is a promising acupuncture improvement method that can regulate immunity and has been widely used in the clinical treatment of a variety of diseases. The aim of this study is to observe the therapeutic effect of CIAA on HCC and to investigate the potential mechanism of immune escape. MATERIALS AND METHODS: A total of 40 mice were randomly divided into three groups: the HCC model group (n = 15), the CIAA treatment group (n = 15), and the control group (n = 10). HCC was chemically induced in 30 mice by the combination of DEN, carbon tetrachloride, and ethanol for 150 days. Among them, 15 were selected for CIAA treatment to ascertain the therapeutic effect. The mRNA expression levels of AFP, IL-10, PD-1, and CTLA-4 in three groups were examined by using RT-PCR. AFP and AKT expressions were measured by using western blotting. PD1, CTLA-4, IL-10, CD4+, and CD8+ protein expression levels were evaluated by using IHC. The mortality rate, body weight, and psychological conditions of three groups were also compared. RESULTS: The mRNA and protein expression levels of AFP, PD-1, CTLA-4, and IL-10 were significantly downregulated in the CIAA-treated mice in comparison with HCC mice. IHC assay shows that CD4+ and CD8+ expression levels were notably upregulated after CIAA treatment. Western blotting assay shows that AKT pathway was deactivated in CIAA-treated mice. CIAA notably reduced the mortality rate and inhibited weight loss caused by HCC and improved the overall psychological condition of the mice. CONCLUSIONS: Taken together, our data corroborate the effective potency of CIAA in the treatment of HCC by and inhibiting immune escape and deactivating the AKT pathway.

Preparation of a multifunctional silver nanoparticles polylactic acid food packaging film using mango peel extract.

As high-efficiency, safe, and low-drug resistant antibacterial agents, silver nanoparticles (AgNPs) have been widely applied in food and biomedicine. AgNPs was prepared using mango peel extract (MPE) as green and cheap reducing agent and stabilizer. In addition, a novel of preservative film material was developed with polylactic acid (PLA) as protective and substrate. AgNPs was characterized by XPS, XRD and TEM, and the size of AgNPs were in the range of 2.5-6.5 nm. The addition of AgNPs improved the mechanical properties of the film and its barrier ability to water vapor and oxygen. The film exhibited excellent antibacterial properties, and the inhibition rate against Escherichia coli and Staphylococcus aureus were above 95%. Furthermore, in terms of safety, the silver migration and cytotoxicity of the film met the relevant standards, and the shelf life of strawberries was significantly extended.

Fabrication of paper-based enzyme immobilized microarray by 3D-printing technique for screening α-glucosidase inhibitors in mulberry leaves and lotus leaves.

BACKGROUND: The discovery of bioactive compounds in traditional Chinese medicine (TCM) has become an important field in TCM modernization. Ligand fishing is a suitable method for discovery of bioactive compounds in complex mixtures such as TCM with high selectivity. Because of unique advantage of low cost and convenience, paper-based microdevices can be good carriers for enzyme immobilized ligand fishing. METHODS: As an important enzyme for glucose metabolism, α-glucosidase was immobilized on polycaprolactone-chitosan-modified paper to prepare the microdevice with unique microfluid structure generated by 3D printing technology, which can be easily applied to screen active compounds in herbal extracts. The preparation conditions of the paper microarray were optimized. The activity of immobilized α-glucosidase was verified by colorimetric reactions which can be easily monitored by cellphone. The paper microarray with α-glucosidase immobilized was used to screen active compounds in the water extracts of mulberry leaves and lotus leaves. RESULTS: Several key parameters including Na2CO3 solution concentration, Na2CO3 solution volume, glutaraldehyde concentration, crosslinking time of glutaraldehyde and time of α-glucosidase immobilization were optimized. The proposed paper-based microarray was successfully applied in screening active compounds in two herbal extracts. Four compounds including chlorogenic acid, quercetin-3-O-glucuronide, isoquercetin, and quercetin were identified as α-glucosidase inhibitors. The compounds with significant non-specific adsorption caused by chitosan, such as isoquercitrin, astragalin, quercetin, were also found to be active compounds. CONCLUSIONS: An enzyme immobilized paper microarray was designed and fabricated in this work. Polycaprolactone and chitosan were used to modify filter paper to prepare paper microarrays. Parameters of paper device preparation were optimized. Our findings suggested that 3D-printing paper-based microarrays can be a simple and low-cost approach for discovery of active compounds of TCM.