Synergistic effects of three traditional herbs green tea,mulberry leaf and corn silk on glucose uptake level of L6 myoblasts and the hypoglycemic mechanism

Background:Green tea,mulberry leaf and corn silk are traditional herbs used in the prevention and treatment of diabetes in China for a long time,but their synergistic hypoglycemic effects and mechanisms remain unclear.Methods:The effective components of green tea,mulberry leaf and corn silk were extracted and enriched.Mixture design of experiments was used to study the influences of different combinations on the cell viability and glucose uptake level of L6 myoblasts,so as to determine the optimal synergistic hypoglycemic combination.The possible hypoglycemic mechanism of the optimal synergistic combination was explored by cytotoxicity assay,glucose uptake assay,and western blot.Results:Three polyphenol enrichment fractions of the herbs,30%ethanol elution fraction of green tea(GT),50%ethanol elution fraction of mulberry leaf(ML)and 60%ethanol elution fraction of corn silk(CS)were obtained.The antioxidant activities of GT-30%,ML-50%and CS-60%were superior to those of crude extracts,and showed strong potential inα-amylase andα-glucosidase inhibition activities.The optimal synergistic combination of crude extracts G7(crude extract of green tea:crude extract of mulberry leaf:crude extract of corn silk=1:5:3),polyphenol enrichment fractions R3(GT-30%:ML-50%:CS-60%=1:7:1)and monomers X2(epigallocatechin gallate:morusin:formononetin=3:1:2)were selected,respectively.G7,R3,and X2 showed promoting effects on the cell viability and glucose uptake of L6 myoblasts within the detected concentration range.In addition,G7,R3,and X2 could increase the expression levels of p-PI3K/PI3K and p-Akt/Akt in L6 myoblasts,and promote the translocation of Glut4,but G7 and R3 showed more significant effects.Conclusion:The synergistic hypoglycemic effects of green tea,mulberry leaf and corn silk had the characteristics of multiple-components and multiple-targets with p-PI3K/PI3K,p-Akt/Akt and the translocation of Glut4 signal pathways involved.The three traditional herbs might have the potential to be combined used for the prevention and treatment of diabetes based on the synergistic hypoglycemic effects.

Insights into a novel nitrogen removal process based on simultaneous anammox and denitrification(SAD) following nitritation with in-situ NOB elimination

Simultaneous anammox and denitrification(SAD) is an efficient approach to treat wastewater having a low C/N ratio;however, few studies have investigated a combination of SAD and partial nitritation(PN). In this study, a lab-scale up-flow blanket filter(UBF) and zeolite sequence batch reactor(ZSBR) were continuously operated to implement SAD and PN advantages, respectively. The UBF achieved a high total nitrogen(TN) removal efficiency of over 70% during the start-up stage(days 1–50), and reached a TN removal efficiency of 96%in the following 90 days(days 51–140) at COD/NH_(4)^(+)-N ratio of 2.5. The absolute abundance of anammox bateria increased to the highest value of 1.58 × 107copies/μL DNA;Comamonadaceae was predominant in the UBF at the optimal ratio. Meanwhile, ZSBR was initiated on day 115 as fast nitritation process to satisfy the influent requirement for the UBF. The combined process was started on day 140 and then lasted for 30 days, during the combined process, between the two reactors, the UBF was the main contributor for TN(66.5% ± 4.5%)and COD(71.8% ± 4.9%) removal. These results demonstrated that strong SAD occurred in the UBF when following a ZSBR with in-situ NOB elimination. This research presents insights into a novel biological nitrogen removal process for low C/N ratio wastewater treatment.

Engineering the thermostability of D-lyxose isomerase from Caldanaerobius polysaccharolyticus via multiple computer-aided rational design for efficient synthesis of D-mannose

D-Mannose is an attractive functional sugar that exhibits many physiological benefits on human health.The demand for low-calorie sugars and sweeteners in foods are increasingly available on the market.Some sugar isomerases,such as D-lyxose isomerase(D-LIase),can achieve an isomerization reaction between D-mannose and D-fructose.However,the weak thermostability of D-LIase limits its efficient conversion from D-fructose to D-mannose.Nonetheless,few studies are available that have investigated the molecular modification of D-LIase to improve its thermal stability.In this study,computer-aided tools including FireProt,PROSS,and Consensus Finder were employed to jointly design D-LIase mutants with improved thermostability for the first time.Finally,the obtained five-point mutant M5(N21G/E78P/V58Y/C119Y/K170P)showed high thermal stability and cat-alytic activity.The half-life of M5 at 65◦C was 10.22 fold,and the catalytic efficiency towards 600 g/L of D-fructose was 2.6 times to that of the wild type enzyme,respectively.Molecular dynamics simulation and intramolecular forces analysis revealed a thermostability mechanism of highly rigidity conformation,newly formed hydrogen bonds andπ-cation interaction between and within protein domains,and redistributed surface electrostatic charges for the mutant M5.This research provided a promising D-LIase mutant for the industrial production of D-mannose from D-fructose.