Identification and Validation of Active Ingredient in Cerebrotein Hydrolysate-I Based on Pharmacokinetic and Pharmacodynamic Studies.

Cerebrotein hydrolysate-1 (CH-1), a mixture of small peptides, polypeptides, and various amino acids derived from porcine brain, has been widely used in the treatment of cerebral injury. However, the bioactive composition and pharmacokinetics of CH-1 are still unexplored because of their complicated composition and relatively tiny amounts in vivo. Herein, NanoLC Orbitrap Fusion Lumos Tribrid Mass Spectrometer was firstly used to qualitatively analyze the components of CH-1. A total of 1347 peptides were identified, of which 43 peptides were characterized by high mass spectrometry (MS) intensity and identification accuracy. We then innovatively synthesized four main peptides for activity verification, and the results suggested that Pep72 (NYEPPTVVPGGDL) had the strongest neuroprotective effect on both in vivo and in vitro models. Next, a quantitative method for Pep72 was established based on liquid chromatography tandem mass spectrometry (LC-MS/MS) with the aid of Skyline software and then used in its pharmacokinetic studies. The results revealed that Pep72 had a high elimination rate and low exposure in rats. In addition, a hCMEC/D3-based in vitro model was built and firstly used to investigate the transport of Pep72. We found that Pep72 had extremely low blood-brain barrier permeability and was not a substrate of efflux transporters. The biotransformation of Pep72 in rat fresh plasma and tissues was investigated to explore the contradiction between pharmacokinetics and efficacy. A total of 11 main metabolites were structurally identified, with PGGDL and EPPTVPGGDL being the main metabolites of Pep72. Notably, metalloproteinase and cysteine protease were confirmed to be the main enzymes mediating Pep72 metabolism in rat tissues. SIGNIFICANCE STATEMENT: The NanoLC Orbitrap Fusion Lumos Tribrid Mass Spectrometer was firstly applied to discover the components of CH-1, and one main peptide Pep72 (NYEPPTVVPGGDL) was innovatively synthesized and firstly found to have the strongest neuroprotective effect among 1347 peptides identified from CH-1. Our study is the first time to identify and verify the active ingredient of CH-1 from the perspective of pharmacokinetics and pharmacodynamics, and provides a systematic technical platforms and strategies for the active substance research of other protein hydrolysates.

Combined multi-band infrared camouflage and thermal management via a simple multilayer structure design.

Infrared camouflage is crucial for high-temperature objects to avoid detection, and spontaneous infrared radiation is also an important way for high-temperature objects to dissipate heat. Therefore, selective infrared emission has become significant for the coating design of surfaces such as aircraft, which require low emission in the atmospheric window band (3-5 µm and 8-14 µm) and high emission outside it (5-8 µm). This Letter employs a simple multilayer film structure to achieve selective regulation of the material emission spectrum. Combining the transfer matrix method and genetic algorithm, a multilayer film structure containing 12 layers of three high-temperature-resistant materials (SiO2, TiO2 and Ge) has been designed. It shows fairly low emissivity in two main bands of infrared detection (ε3∼5µm=0.14, ε8∼14µm=0.21) and high emissivity outside them (ε5∼8µm=0.86), and this infrared selectivity can be well maintained with the incident angle rising from 0 to 60 deg. The Poynting vector distribution in the material at different incident wavelengths is analyzed to further explore the interference mechanism to achieve spectral selective emission. The significance of this work lies in the construction of a relatively simple coating design while ensuring efficient infrared camouflage and thermal management performance.

Insights into Q-markers and molecular mechanism of Sanguisorba saponins in treating ulcerative colitis based on lipid metabolism regulation.

BACKGROUND: Sanguisorba saponin extract (SSE) is the main active part of Sanguisorba officinalis with various pharmacological activities such as anti-inflammatory, anti-bacterial and anti-oxidant. However, its therapeutic role and underlying mechanisms for ulcerative colitis (UC) still need to be elucidated. PURPOSE: This study aims to explore the therapeutic effect, effectiveness-material basis-quality markers (Q-markers) and prospective mechanism of function of SSE on UC. METHODS: Fresh 2.5% dextran sulfate sodium salt (DSS) solution was placed in drinking bottles for 7 days to induce a mouse model of UC. SSE and sulfasalazine (SASP) were supplemented to mice by gavage for consecutive 7 days to investigate the therapeutic role of SSE on UC. Mouse monocyte macrophages (RAW264.7) and human normal colonic epithelial (NCM460) cells were treated with LPS to induce inflammatory responses, followed by pharmacodynamic examination with different concentrations of SSE. Hematoxylin-eosin (HE) and Alcian blue staining were conducted to evaluate the pathological damage of mice colon. Lipidomic technology was conducted to explore the differential lipids closely related to the disease process of UC. Quantitative PCR analysis, immunohistochemistry and ELISA kit were used to measure the expression levels of the corresponding proteins and pro-inflammatory factors. RESULTS: SSE treatment could effectively reduce the elevated expressions of pro-inflammatory factors in RAW264.7 and NCM460 cells due to LPS stimulation. Intragastric administration of SSE was found to significantly alleviate the symptoms of DSS-induced colon injury and low-polar saponins in SSE. Low polarity saponins, especially ZYS-II, were proved to be the main active substances of SSE in treating UC. In addition, SSE could significantly ameliorate the aberrant lipid metabolism in UC mice. The role of phosphatidylcholine (PC)34:1 in the UC pathogenesis has been fully verified in our previous studies. Herein, SSE-dosing effectively reversed the metabolic disorder of PCs in UC mice, and increased the PC34:1 level to normal via up-regulating the expression of phosphocholine cytidylyltransferase (PCYT1α). CONCLUSION: Our data innovatively revealed that SSE could significantly alleviate the symptoms of UC by reversing the disorder of PC metabolism induced by DSS modeling. SSE was proved for the first time to be a promising and effective candidate for UC treatment.

Schisandra lignans ameliorate nonalcoholic steatohepatitis by regulating aberrant metabolism of phosphatidylethanolamines.

Nonalcoholic steatohepatitis (NASH) is a spectrum of chronic liver disease characterized by hepatic lipid metabolism disorder. Recent reports emphasized the contribution of triglyceride and diglyceride accumulation to NASH, while the other lipids associated with the NASH pathogenesis remained unexplored. The specific purpose of our study was to explore a novel pathogenesis and treatment strategy of NASH via profiling the metabolic characteristics of lipids. Herein, multi-omics techniques based on LC-Q-TOF/MS, LC-MS/MS and MS imaging were developed and used to screen the action targets related to NASH progress and treatment. A methionine and choline deficient (MCD) diet-induced mouse model of NASH was then constructed, and Schisandra lignans extract (SLE) was applied to alleviate hepatic damage by regulating the lipid metabolism-related enzymes CES2A and CYP4A14. Hepatic lipidomics indicated that MCD-diet led to aberrant accumulation of phosphatidylethanolamines (PEs), and SLE could significantly reduce the accumulation of intrahepatic PEs. Notably, exogenous PE (18:0/18:1) was proved to significantly aggravate the mitochondrial damage and hepatocyte apoptosis. Supplementing PE (18:0/18:1) also deteriorated the NASH progress by up regulating intrahepatic proinflammatory and fibrotic factors, while PE synthase inhibitor exerted a prominent hepatoprotective role. The current work provides new insights into the relationship between PE metabolism and the pathogenesis of NASH.

Strategies for mapping protein hydrolysate profiles and pharmacokinetics based on non-targeted proteomics combining skyline-aided quantitative techniques.

Numerous works have been focused on the bioactivities of protein hydrolysates (PHs) and their application in food or drug formulations, but their composition and pharmacokinetics have never been addressed due to their complex constitutes, short half-life, extremely low concentrations and lack of authentic standards. The present study aims to develop systematic analytical strategy and technical platform with optimized sample preparation, separation and detection protocols for PHs. Lineal peptides (LPs), extraction of the spleen of healthy pigs or calves, were used as cases. First, solvents with polarity gradients were used to globally extract peptides of LP from biological matrix. Non-targeted proteomics based on a high-resolution MS system was used to establish a reliable qualitative analysis workflow for PHs. Based on the developed approach, 247 unique peptides were identified using NanoLC-Orbitrap-MS/MS, and then further verified on the MicroLC-Q-TOF/MS system. In the quantitative analysis workflow, Skyline software was used to predict and optimize the LC-MS/MS detection parameters of LPs followed by investigating the linearity and precision of the developed analytical assay. Note worthily, we innovatively prepared calibration curves by sequential dilution of LP solution to overcome the bottleneck of lacking authentic standards and complex PH composition. All the peptides exhibited good linearity and precision in biological matrix. The established qualitative and quantitative assays were successfully applied to study the distribution characteristics of LPs in mice, and would be conductive to systematically map the profile and pharmacokinetics of peptides in various PHs in vivo and in vitro.

Experimental Study on Capillary Microflows in High Porosity Open-Cell Metal Foams.

Metal foams have been widely used in heat pipes as wicking materials. The main issue with metal foams is the surface property capillary limit. In this paper, a chemical blackening process for creating a superhydrophilic surface on copper foams is studied with seven different NaOH and NaClO2 solution concentrations (1.5~4.5 mol/L), in which the microscopic morphology of the treated copper foam surface is analyzed by scanning electron microscopy. The capillary experiments are carried out to quantify the wicking characteristics of the treated copper foams and the results are compared with theoretical models. A the microscope is used to detect the flow stratification characteristics of the capillary rise process. The results show that the best wicking ability is obtained for the oxidation of copper foam using 3.5 mol/L of NaOH and NaClO2 solution. Gravity plays a major role in defining the permeability and effective pore radius, while the effect of evaporation can be ignored. The formation of a fluid stratified interface between the unsaturated and saturated zone results in capillary performance degradation. The current study is important for understanding the flow transport in porous materials.

Human dendritic cell targeting peptide can be targeted to porcine dendritic cells to improve antigen capture efficiency to stimulate stronger immune response.

Dendritic cells (DCs) play a key role in the natural recognition of pathogens and subsequent activation of adaptive immune responses due to their potent antigen-presenting ability. Dendritic cell-targeting peptide (DCpep) is strongly targeted to DCs, which often express antigens, to enhance the efficacy of vaccines. Our previous study showed that recombinant Lactobacillus expressing human DCpep could significantly induce stronger immune responses than recombinant Lactobacillus without DCpep, but the mechanism remains unclear. In this study, the mechanism by which DCpep enhances the immune response against recombinant Lactobacillus was explored. Fluorescence-labeled human DCpep was synthesized to evaluate the binding ability of human DCpep to porcine monocyte-derived dendritic cells (Mo-DCs) and DCs of the small intestine. The effects of Mo-DC function induced by recombinant Lactobacillus expressing human DCpep fused with the porcine epidemic diarrhea virus (PEDV) core neutralizing epitope (COE) antigen were also investigated. The results showed that human DCpep bind to porcine DCs, but not to porcine small intestinal epithelial cells. Human DCpep can also improve the capture efficiency of recombinant Lactobacillus by Mo-DCs, promote the maturation of dendritic cells, secrete more cytokines, and enhance the ability of porcine DCs to activate T-cell proliferation. Taken together, these results promote advanced understanding of the mechanism by which DCpep enhances immune responses. We found that some DCpeps are conserved between humans and pigs, which provides a theoretical basis for the development of a DC-targeted vaccine.

Femtosecond Laser Fabrication of Micro and Nano-Structures on CIGS/ITO Bilayer Films for Thin-Film Solar Cells.

Cu(In, Ga)Se2 (CIGS) thin films have attracted considerable interest as potential photovoltaic solar cells. Moreover, several current studies are focusing on improving their conversion efficiency. This study proposes a method to process micro- and nanostructures onto the surface of CIGS/ITO bilayer films to broaden the field of solar cell application. The bilayer films exhibited optical characteristics different from those of a single-film during processing. Field intensities at different layer positions of the CIGS/ITO bilayer films were analyzed, and different structures were fabricated by varying a set of parameters. Ripples were obtained using a pulse energy of 0.15 µJ and scanning speeds in the range of 0.1-1 mm/s, but after increasing speed to 3-5 mm/s, ripple structures were produced that had a large period of several microns and spatial porous nanostructures. This pattern exhibited low reflectivity. Optimal structures were obtained at a scanning speed of 3.5 mm/s a pulse energy of 0.15 µJ, and a reflectivity lower than 5%. Large areas characterized by micron-sized ripple structures and accompanied by nanoscale porous structures presented high optical performance and efficiency, which can be used to broaden the application of thin film-based solar cells.

Review and a Theoretical Approach on Pressure Drop Correlations of Flow through Open-Cell Metal Foam.

Due to their high porosity, high stiffness, light weight, large surface area-to-volume ratio, and excellent thermal properties, open-cell metal foams have been applied in a wide range of sectors and industries, including the energy, transportation, aviation, biomedical, and defense industries. Understanding the flow characteristics and pressure drop of the fluid flow in open-cell metal foams is critical for applying such materials in these scenarios. However, the state-of-the-art pressure drop correlations for open-cell foams show large deviations from experimental data. In this paper, the fundamental governing equations of fluid flow through open-cell metal foams and the determination of different foam geometry structures are first presented. A variety of published models for predicting the pressure drop through open-cell metal foams are then summarized and validated against experimental data. Finally, two empirical correlations of permeability are developed and recommended based on the model of Calmidi. Moreover, Calmidi's model is proposed to calculate the Forchheimer coefficient. These three equations together allow calculating the pressure drop through open-cell metal foam as a function of porosity and pore diameter (or strut diameter) in a wide range of porosities ε = 85.7-97.8% and pore densities of 10-100 PPI. The findings of this study greatly advance our understanding of the flow characteristics through open-cell metal foam and provide important guidance for the design of open-cell metal foam materials for different engineering applications.

Design, synthesis and biological evaluation of novel 2-phenyl pyrimidine derivatives as potent Bruton's tyrosine kinase (BTK) inhibitors.

BTK is an effective target for the treatment of B-cell malignant tumors and autoimmune diseases. In this work, a series of 2-phenyl pyrimidine derivatives were prepared and their preliminary in vitro activities on B-cell leukemia cells as well as the BTK enzyme were determined. The results showed that compound 11g displayed the best inhibitory activity on BTK with an inhibition rate of 82.76% at 100 nM and excellent anti-proliferation activity on three B-cell leukemia lines (IC50 = 3.66 µM, 6.98 µM, and 5.39 µM against HL60, Raji and Ramos, respectively). Besides, the flow cytometry analysis results indicated that 11g inhibited the proliferation of the Raji cells in a dose- and time-dependent manner, and blocked the Ramos cells at the G0/G1 phase, which is in accordance with the positive control ibrutinib. The mechanism investigation demonstrated that 11g could inhibit the phosphorylation of BTK and its downstream substrate phospholipase γ2 (PLCγ2). All these results showed that 11g was a promising lead compound that merited further optimization as a novel class of BTK inhibitor for the treatment of B-cell lymphoblastic leukemia.

Optimization of novel benzofuro[3,2-b]pyridin-2(1H)-one derivatives as dual inhibitors of BTK and PI3Kδ.

BTK and PI3Kδ play crucial roles in the progression of leukemia, and studies confirmed that the dual inhibition against BTK and PI3Kδ could provide superior anticancer agents to single targeted therapies. Herein, a new series of novel benzofuro[3,2-b]pyridin-2(1H)-one derivatives were optimized based on a BTK/PI3Kδ inhibitor 2 designed by our group. Biological studies clarified that compound 6f exhibited the most potent inhibitory activity (BTK: IC50 = 74 nM; PI3Kδ: IC50 = 170 nM) and better selectivity than 2. Moreover, 6f significantly inhibited the proliferation of Raji and Ramos cells with IC50 values of 2.1 µM and 2.65 µM respectively by blocking BTK and PI3K signaling pathways. In brief, 6f possessed of the potency for further optimization as an anti-leukemic drug by inhibiting BTK and PI3Kδ kinase.