Puerarin Induces Macrophage M2 Polarization to Exert Antinonalcoholic Steatohepatitis Pharmacological Activity via the Activation of Autophagy.

To determine the protective mechanism of puerarin against nonalcoholic steatohepatitis (NASH), the pharmacodynamic effects of puerarin on NASH were evaluated by using zebrafish, cells, and mice. Western blotting, flow cytometry, immunofluorescence, and qRT-PCR were used to detect the effects of puerarin on RAW264.7 autophagy and polarization. Key target interactions between autophagy and polarization were detected using immunoprecipitation. Puerarin regulated the M1/M2 ratio of RAW 264.7 cells induced by LPS + INF-γ. Transcriptomics revealed that PAI-1 is a key target of puerarin in regulating macrophage polarization. PAI-1 knockout reduced the number of M1-type macrophages and increased the number of M2-type macrophages. Puerarin regulated PAI-1 and was associated with macrophage autophagy. It increased p-ULK1 expression in macrophages and activated autophagic flux, reducing the level of PAI-1 expression. Stat3/Hif-1α and PI3K/AKT signaling pathways regulated the number of macrophage polarization phenotypes, reducing liver lipid droplet formation, alleviating liver structural abnormalities, decreasing the number of cytoplasmic vacuoles, and decreasing the area of blue collagen in NASH mice. Puerarin is a promising dietary component for NASH alleviation.

American Ginseng for the Treatment of Alzheimer's Disease: A Review.

Alzheimer's disease (AD) is a prevalent degenerative condition that is increasingly affecting populations globally. American ginseng (AG) has anti-AD bioactivity, and ginsenosides, as the main active components of AG, have shown strong anti-AD effects in both in vitro and in vivo studies. It has been reported that ginsenosides can inhibit amyloid ß-protein (Aß) production and deposition, tau phosphorylation, apoptosis and cytotoxicity, as well as possess anti-oxidant and anti-inflammatory properties, thus suppressing the progression of AD. In this review, we aim to provide a comprehensive overview of the pathogenesis of AD, the potential anti-AD effects of ginsenosides found in AG, and the underlying molecular mechanisms associated with these effects. Additionally, we will discuss the potential use of AG in the treatment of AD, and how ginsenosides in AG may exert more potent anti-AD effects in vivo may be a direction for further research.

Preventive and therapeutic effects of natural products and herbal extracts on nonalcoholic fatty liver disease/nonalcoholic steatohepatitis.

Nonalcoholic fatty liver disease (NAFLD) is a common condition that is prevalent in patients who consume little or no alcohol, and is characterized by excessive fat accumulation in the liver. The disease is becoming increasingly common with the rapid economic development of countries. Long-term accumulation of excess fat can lead to NAFLD, which represents a global health problem with no effective therapeutic approach. NAFLD is a complex, multifaceted pathological process that has been the subject of extensive research over the past few decades. Herbal medicines have gained attention as potential therapeutic agents to prevent and treat NAFLD due to their high efficacy and low risk of side effects. Our overview is based on a PubMed and Web of Science database search as of Dec 22 with the keywords: NAFLD/NASH Natural products and NAFLD/NASH Herbal extract. In this review, we evaluate the use of herbal medicines in the treatment of NAFLD. These natural resources have the potential to inform innovative drug research and the development of treatments for NAFLD in the future.

Renal function protection and the mechanism of ginsenosides: Current progress and future perspectives.

Nephropathy is a general term for kidney diseases, which refers to changes in the structure and function of the kidney caused by various factors, resulting in pathological damage to the kidney, abnormal blood or urine components, and other diseases. The main manifestations of kidney disease include hematuria, albuminuria, edema, hypertension, anemia, lower back pain, oliguria, and other symptoms. Early detection, diagnosis, and active treatment are required to prevent chronic renal failure. The concept of nephropathy encompasses a wide range of conditions, including acute renal injury, chronic kidney disease, nephritis, renal fibrosis, and diabetic nephropathy. Some of these kidney-related diseases are interrelated and may lead to serious complications without effective control. In serious cases, it can also develop into chronic renal dysfunction and eventually end-stage renal disease. As a result, it seriously affects the quality of life of patients and places a great economic burden on society and families. Ginsenoside is one of the main active components of ginseng, with anti-inflammatory, anti-tumor, antioxidant, and other pharmacological activities. A variety of monomers in ginsenosides can play protective roles in multiple organs. According to the difference of core structure, ginsenosides can be divided into protopanaxadiol-type (including Rb1, Rb3, Rg3, Rh2, Rd and CK, etc.), and protopanaxatriol (protopanaxatriol)- type (including Rg1, Rg2 and Rh1, etc.), and other types (including Rg5, Rh4, Rh3, Rk1, and Rk3, etc.). All of these ginsenosides showed significant renal function protection, which can reduce renal damage in renal injury, nephritis, renal fibrosis, and diabetic nephropathy models. This review summarizes reports on renal function protection and the mechanisms of action of these ginsenosides in various renal injury models.

Exploring the potential of ginseng glycoprotein to improve learning and memory in mice via Notch signaling pathway and structural analysis using multi-information fusion based on liquid chromatography-mass spectrometry.

ETHNOPHARMACOLOGICAL RELEVANCE: Panax ginseng C.A. Meyer reportedly exhibits various beneficial pharmacological activities. Panax ginseng glycoproteins (PGG) are a class of glycosylated protein components extracted from ginseng and can exert significant activity for improving learning and memory abilities. AIM OF THE STUDY: The objective of the present study was to investigate the PGG-mediated protective mechanism against neurodegenerative diseases via the Notch signaling pathway using proteomic methods. MATERIALS AND METHODS: We examined learning and memory in mice using the Morris water maze and nest-building paradigms. The PGG structure was determined using multi-information fusion based on liquid chromatography-mass spectrometry (LC/MS). Accurate glycosylation sites of glycoproteins were identified using the advanced glycosylation analysis software Byonic. Furthermore, connection modes of the oligosaccharide chain were clarified by methylation analysis of sugar residues. The differentially expressed proteins (DEPs) between wild-type (WT) and APP/APS1 mice were measured and compared using label-free quantitative proteomics, and related signaling pathways were identified. For validation, we performed a series of in vitro tests, including an assessment of cell viability, apoptosis assay, quantitative real-time polymerase chain reaction, and western blotting. RESULTS: In the Morris water maze and nesting experiments, PGG-treated WT mice exhibited significantly improved learning and memory. The structures of 171 glycoprotein fragments in PGG matched the credible score, and typical structures were identified using LC/MS data analysis. According to the proteomic analysis results, 188 DEPs were detected between the model and administration groups, and two downregulated DEPs were related to the Notch signaling pathway. Based on the in vitro verification tests, PGG significantly inhibited the expression of key proteins in the Notch signaling pathway in microglia. CONCLUSIONS: PGG could prevent the development of neuroinflammation by inhibiting excessive activation of the Notch signaling pathway, thereby inhibiting neuroapoptosis.

Studies on Chemical Composition of Pueraria lobata and Its Anti-Tumor Mechanism.

Fourteen compounds were isolated from Pueraria lobata (Willd.) Ohwi by column chromatography and preparative thin-layer chromatography; the structures were identified by spectroscopic analysis and compared with data reported in the literature. Seven compounds were isolated and identified from Pueraria lobata for the first time: Linoleic acid, Sandwicensin, Isovanillin, Ethyl ferulate, Haginin A, Isopterofuran, 3'.7-Dihydroxyisoflavan. The other 10 compounds were structurally identified as follows: Lupenone, Lupeol, ß-sitosterol, Genistein, Medicarpin, Coniferyl Aldehyde, Syringaldehyde. All compounds were evaluated for their ability to inhibit SW480 and SW620 cells using the CCK-8 method; compound 5 (Sandwicensin) had the best activity, and compounds 6, 9, 11 and 12 exhibited moderate inhibitory activity. In addition, the targets and signaling pathways of Sandwicensin treatment for CRC were mined using network pharmacology, and MAPK3, MTOR, CCND1 and CDK4 were found to be closely associated with Sandwicensin treatment for CRC; the GO and KEGG analysis showed that Sandwicensin may directly regulate the cycle, proliferation and apoptosis of CRC cells through cancer-related pathways.

Structural analysis of Panax ginseng glycoproteins and its anti-oligoasthenozoospermia effect in vivo.

A component from ginseng in which sugars and proteins are covalently bound is named Panax ginseng glycoproteins (PGG). The contents of neutral carbohydrate, acid carbohydrate, and protein were 45.4%, 4.3% and 51.1%. The average molecular weight was 12,690 Da. The structure analysis showed that PGG had more than 1100 glycoproteins with molecular weight between 308.13 Da and 9991.52 Da, it was divided into two parts: long chain structure and short chain structure. These two parts were compared in molecular mass, number of amino acids, theoretical pI, instability index, aliphatic index and GRAVY. The in vivo distribution test of mice showed that PGG was enriched in mice testis, testicular tissue sections showed strong fluorescence signal expression on the surface of seminiferous tubules. We used cyclophosphamide (CP) to establish a mice model of oligoasthenozoospermia to investigate the anti-oligoasthenozoospermic effect of PGG. The results showed that PGG increased the levels of sex hormones T, FSH, PRL and sperm quality. Histopathology demonstrated that PGG promoted the differentiation process. The organ coefficient indicated that PGG had no obvious toxic and side effects. And the mechanism may be to affect the expression of protein levels such as p-ERK/ERK, p-AKT/AKT, Caspase-3, Bcl-2 and Bax. Therefore, PGG has the potential to develop into drugs for improving spermatogenic disorders.

Structure-activity relationship analysis of Panax ginseng glycoproteins with cytoprotective effects using LC-MS/MS and bioinformatics.

Panax ginseng glycoproteins (PGG) has been shown biological activity, but researches in this field are rarely reported. In this paper, PGG were prepared by reflux and then purified with macroporous resin column. Further separation and purification of PGG using high performance liquid chromatography (HPLC) and two major components (PGG-1, PGG-2) were obtained. The molecular weights were calculated by gel permeation chromatography (GPC), and the results are 1.5 KDa and 8.2 KDa respectively. The MTT assay was used to study the cytoprotective effects of PGG, the results exhibited that PGG had significant effect (P < 0.01), and showed an obvious dose-effect relationship. Anti-apoptosis experiment results showed that PGG and PGG-2 can inhibit Aß-induced apoptosis in SH-SY5Y cells (P < 0.05), and PGG-2 displayed better activity. The structures of N- and O-glycan were determined by combination of LC-MS/MS and methylation analysis. The computed parameters of PGG determined by MS including the theoretical isoelectric point (pI), instability index, aliphatic index and grand average of hydropathicity (GRAVY) were summarized systematically. The distinct differences between two parts would affect the behavior of PGG in vivo. The results of activity test and bioinformatics analysis would guide the study of PGG in pharmacokinetics and mechanism.

LC-MS/MS analysis of partial structure of Panax ginseng protein and its distribution in vivo.

Protein from Panax ginseng can improve learning, memory, and analgesia. Here, we investigated a fluorescence labeling method that can be used to determine the in vivo distribution of P. ginseng protein (PGP). High-performance liquid chromatography (HPLC) was used to define the amino acid composition and molecular weight of PGP; LC-MS/MS was used to identify the PGP structure, which was fluorescently-labeled using a fluorescein isothiocyanate (FITC) probe. The connection form of the PGP fluorescent marker (PGP-FITC) was identified by ultraviolet and infrared spectrophotometry. The in vivo distribution of PGP was observed by fluorescence imaging, and tissue content was determined. Results showed that PGP was enriched in the brain and that vascular epithelial cells showed specific uptake. We provide an experimental method to label and identify the in vivo distribution of PGP, which forms the basis for future studies to determine whether PGP can penetrate the blood-brain barrier (BBB) and elucidate the transport mechanism.

Metabonomics Study of Ginseng Glycoproteins on Improving Sleep Quality in Mice.

The changes of brain metabolism in mice after injection of ginseng glycoproteins (GPr) were analyzed by gas chromatography mass spectrometry- (GC/MS-) based metabolomics platform. The relationship between sedative and hypnotic effects of ginseng glycoproteins and brain metabolism was discussed. Referring to pentobarbital sodium subthreshold test, we randomly divided 20 mice into two groups: control and ginseng glycoproteins group. The mice from the control group were treated with normal saline by i.p and GPr group were treated with 60 mg/kg of GPr by i.p. The results indicated that GPr could significantly improve the sleep quality of mice. Through multivariate statistical analysis, we found that there were 23 differential metabolites in whole brain tissues between the control group and the GPr group. The pathway analysis exhibited that GPr may be involved in the regulation of the pathway including purine metabolism, nicotinate and nicotinamide metabolism, glycine, serine and threonine metabolism, arginine and proline metabolism, alanine, aspartate and glutamate metabolism, and steroid hormone biosynthesis. This work is helpful to understand the biochemical mechanism of GPr on promoting sleep and lay a foundation for further development of drugs for insomnia.

Study on the Structure of Ginseng Glycopeptides with Anti-Inflammatory and Analgesic Activity.

Panax ginseng is well known for its medicinal functions. As a class of important compound of ginseng, ginsenoside is widely studied around the world. In addition, ginseng glycopeptides also showed good biological activity, but researches in this field are rarely reported. In this study, ginseng glycopeptides (Gg) were first prepared from Panax ginseng by reflux extracted with 85% ethanol and the following purification with Sephadex G-15 column. Then, the inflammatory pain models induced by carrageenan and the rat pain models induced by Faure Marin were established for research on mechanism of analgesic activities. It is showed that Gg had an obvious inhibiting effect on inflammation and a significant reduction on the Malondialdehyde (MDA) of inflammatory foot tissue. And there were significant differences between moderate to high dose of Gg and model group in Interleukin 1ß (IL-1ß), Interleukin 2 (IL-2), Interleukin 4 (IL-4), Tumor necrosis factor α (TNF-α) and Histamine. The two models can be preliminarily determined that the analgesic effect of Gg may be peripheral, which mechanism may be related to the dynamic balance between proinflammatory cytokines (TNF-α, IL-1ß) and anti-inflammatory cytokines (IL-2, IL-4, and Interleukin 10 (IL-10)). A series of methods were used to study Gg in physical-chemical properties and linking mode of glycoside. The high-resolution mass spectrometry was used for identification of the structure of Gg. Moreover, the structure of 20 major Gg were investigated and identified. The structural analysis of Gg was benefit for the next study on structure-activity relationship.

In vivo and in vitro neuroprotective effects of Panax ginseng glycoproteins.

The root of Panax ginseng C. A. Mey (Araliaceae) has medicinal value in complex system of Traditional Chinese medicines for its use in improving cognitive function. A glycoproteins named PGL-1 was extracted from ginseng which subjected to through a macroporous resin, hollow-fiber ultrafiltration and dialyzed. The glycoproteins has a molecular weight in the range from 0.4 to 4.4kDa, with an average molecular mass of 1.6kDa. HPLC analysis revealed that the compositions of glycoproteins included fucose, mannose, rhamnose, glucose, galacturonic acid, N-acetylglucosamine and N-acetylgalactosamine. Glycan of PGL-1 has a backbone of →4)-Rha-(1→, →4)-Fuc -(1→, →6)-Gal-(1→, →4)-GalA-(1→, →4)-GlcNAc-(1→ and →4)-GalNAc-(1→,and (→3,6)-Man-(1→) was distributed in branches. The (1→)-Fuc, (1→)-Glc and (1→)-GlcNAc or (1→)-GalNAc were regarded as a terminal residue. The Morris water maze test revealed that the PGL-1 can effectively alleviate the memory impairment symptoms of rats induced by Aß25-35. All dose groups showed significant activity of protective effect on apoptosis SH-SY5Y induced by Aß25-35, and obviously inhibited the S phase arrest. Compared with Aß25-35 treatment alone, a significant reduction in NO concentration and NOS activity was detected in cells co-administered with glycoproteins. Thus, glycoproteins derived from ginseng might be a promising anti-AD reagent.

Fabrication of heterogeneous double-ring-like structure arrays by combination of colloidal lithography and controllable dewetting.

We report a novel technique for fabricating the heterogeneous double-ring-like structural array by colloidal lithography and two-step dewetting process. First, the 2D non-closed-packed (ncp) silica sphere arrays were obtained by lift-up lithography. Then, the ncp sphere array transferred onto the Rhodamine B (RB)@poly(vinyl alcohol) (PVA) film was used for the mask during reactive ion etching (RIE) process. Sequentially, the substrate with RB@PVA ring-like structure arrays under the silica sphere was dip-coated from poly(N-vinylcarbazole) (PVK) chloroform solution with certain concentration. Due to the presence of ordered 2D sphere arrays, the two-step dewetting behavior happened on top of the sphere and the silicon wafer between adjacent spheres, respectively. After removing the silica sphere arrays by hydrofluoric acid, the RB@PVA/PVK heterogeneous double-ring-like structure array was exhibited on the substrate. We characterized this particular structure by SEM, AFM, and fluorescence spectrum, which prove that both the inner RB@PVA ring and outer PVK ring are independent without any reaction. Accordingly, this method could be extended to other materials owing to its universality. These unique structural arrays have potential application in optoelectronic devices, surface photocatalysis, and surface enhanced Raman scattering (SERS).

A universal approach to fabricate ordered colloidal crystals arrays based on electrostatic self-assembly.

We present a novel and simple method to fabricate two-dimensional (2D) poly(styrene sulfate) (PSS, negatively charged) colloidal crystals on a positively charged substrate. Our strategy contains two separate steps: one is the three-dimensional (3D) assembly of PSS particles in ethanol, and the other is electrostatic adsorption in water. First, 3D assembly in ethanol phase eliminates electrostatic attractions between colloids and the substrate. As a result, high-quality colloidal crystals are easily generated, for electrostatic attractions are unfavorable for the movement of colloidal particles during convective self-assembly. Subsequently, top layers of colloidal spheres are washed away in the water phase, whereas well-packed PSS colloids that are in contact with the substrate are tightly linked due to electrostatic interactions, resulting in the formation of ordered arrays of 2D colloidal spheres. Cycling these processes leads to the layer-by-layer assembly of 3D colloidal crystals with controllable layers. In addition, this strategy can be extended to the fabrication of patterned 2D colloidal crystals on patterned polyelectrolyte surfaces, not only on planar substrates but also on nonplanar substrates. This straightforward method may open up new possibilities for practical use of colloidal crystals of excellent quality, various patterns, and controllable fashions.

Controlled fabrication of fluorescent barcode nanorods.

We report a novel technique for generating polymer fluorescent barcode nanorods by reactive ion etching of polymer multilayer films using nonclose-packed (ncp) colloidal microsphere arrays as masks. The fluorescent polymer multilayer films were spin-coated on a substrate, and ncp microsphere arrays were transferred onto these films. The exposed polymers were then etched away selectively, leaving color-encoded nanorods with well-preserved fluorescent properties. By modifying the spin-coating procedure, the amount of polymer in each layer could be tuned freely, which determined the relative fluorescence intensity of the barcode nanorods. These nanorod arrays can be detached from the substrate to form dispersions of coding materials. Moreover, the shape of the nanorods is controllable according to the different etching speeds of various materials, which also endows the nanorods with shape-encoded characters. This method offers opportunities for the fabrication of novel fluorescent barcodes which can be used for detecting and tracking applications.

Elliptical silicon arrays with anisotropic optical and wetting properties.

We demonstrate a facile etching method to fabricate silicon elliptical pillar arrays (Si-EPAs) with unique anisotropic optical and wetting characters using polystyrene elliptical hemisphere arrays (EHAs) as mask. The EHAs were fabricated via a modified micromolding method. By varying the experimental conditions in the fabrication process, the morphology of the resulting microstructures can be controlled exactly. Because of the anisotropic morphology of the elliptical pillar, the Si-EPA shows unique anisotropic properties, such as anisotropic surface reflection and anisotropic wetting property. Additionally, through oblique evaporation deposition of Au and selective chemical modification to turn the elliptical pillars into "Janus" elliptical pillars, the "Janus" Si-EPA shows more peculiar anisotropic properties owing to the further increased asymmetry. We believe that the Si-EPAs will have potential applications in anisotropic optical and electronic devices.

Mimicking the rice leaf--from ordered binary structures to anisotropic wettability.

In this paper, we report a method to fabricate a series of surfaces with large-area ordered binary arrays by controllable dewetting. The binary structure arrays consist of an ordered-stripe array and droplet-row array. In order to expand the system, polystyrene (PS) and poly(methyl methacrylate) (PMMA) are introduced in this experiment for investigation in detail. Through adjustment of the polymer solution concentration and the modified underlying pattern on substrate, the surface topographies can be controlled simply. Accordingly, three types of topographies with ordered binary arrays have been obtained by thermal annealing. These unique surfaces mimic the natural rice leaf structurally, which also displays anisotropic wettability for water droplet as natural surfaces. This method points out a new way for the manufacture of functional surfaces.

A versatile approach to fabricate ordered heterogeneous bull's-eye-like microstructure arrays.

In this paper, ordered heterogeneous bull's-eye-like microstructure arrays were fabricated through a simple two-step method on gold substrate with patterned self-assemble monolayers (SAMs). First, we prepared ordered polymer dot arrays on the SAMs patterned gold substrate by SAMs-direct dewetting. Subsequently, by manipulating concentration-controlled dewetting process, ordered ring arrays were obtained on the dot arrays patterned surface under the protection of water droplets. Namely, ordered bull's-eye-like structure arrays were fabricated successfully. The mechanism of these two kinds of dewetting process has been investigated in detail. And due to these two steps were independent, different materials could be simply introduced to the current system. Therefore, ordered homogeneous and heterogeneous bull's-eye-like structure arrays such as poly(N-vinylcarbazole) (PVK) (dot)/PVK (ring), PVK/5,12-ditetradecylquinolino[2,3-b]acridine-7,14(5H,12H)-dione (DTQA), and PVK/Fe(3)O(4) nanoparticles were obtained. This straightforward method may open up new possibilities for practical use of microchips with binary heterogeneous structure arrays.

Modulating two-dimensional non-close-packed colloidal crystal arrays by deformable soft lithography.

We report a simple method to fabricate two-dimensional (2D) periodic non-close-packed (ncp) arrays of colloidal microspheres with controllable lattice spacing, lattice structure, and pattern arrangement. This method combines soft lithography technique with controlled deformation of polydimethylsiloxane (PDMS) elastomer to convert 2D hexagonal close-packed (hcp) silica microsphere arrays into ncp ones. Self-assembled 2D hcp microsphere arrays were transferred onto the surface of PDMS stamps using the lift-up technique, and then their lattice spacing and lattice structure could be adjusted by solvent swelling or mechanical stretching of the PDMS stamps. Followed by a modified microcontact printing (microcp) technique, the as-prepared 2D ncp microsphere arrays were transferred onto a flat substrate coated with a thin film of poly(vinyl alcohol) (PVA). After removing the PVA film by calcination, the ncp arrays that fell on the substrate without being disturbed could be lifted up, deformed, and transferred again by another PDMS stamp; therefore, the lattice feature could be changed step by step. Combining isotropic solvent swelling and anisotropic mechanical stretching, it is possible to change hcp colloidal arrays into full dimensional ncp ones in all five 2D Bravais lattices. This deformable soft lithography-based lift-up process can also generate patterned ncp arrays of colloidal crystals, including one-dimensional (1D) microsphere arrays with designed structures. This method affords opportunities and spaces for fabrication of novel and complex structures of 1D and 2D ncp colloidal crystal arrays, and these as-prepared structures can be used as molds for colloidal lithography or prototype models for optical materials.

Morphology and wettability control of silicon cone arrays using colloidal lithography.

In this paper we present a simple method to fabricate ordered silicon cone arrays with controllable morphologies on a silicon substrate using reactive ion etching with two-dimensional silica colloidal crystals as masks. The etching process and the morphologies of the obtained structure are quantified. Unlike works reported previously, we show that the surface roughness of the obtained silicon cone arrays can be adjusted by controlling the etching duration, which is proved to be of importance in tailoring the behavior of water droplets when being used as antireflection coatings with superhydrophobicity. Moreover, this strategy is compatible with the methods we have established on controlling the arrangement of colloidal spheres, and thus silicon cone arrays with tunable periodicities, different lattice structures, and various patterns can be prepared. The obtained silicon cone arrays with strips can be used as hydrophobic substrates with anisotropic dewetting just like the leaves of rice. It is found that by adjusting the strip width with and without silicon cones, the water droplets can transform from isotropic dewetting to anisotropic dewetting.