Assessment of the Effects of Structural Modification of Gastrodia elata Polysaccharide on Anti-Breast Cancer Activity Using Asymmetrical Flow Field-Flow Fractionation.

Gastrodia elata ("Tian Ma" in Chinese) is used as a food and medical ingredient in traditional Chinese medicine. In this study, to enhance the anti-breast cancer activity of Gastrodia elata polysaccharide (GEP), GEPs were modified via sulfidation (SGEP) and acetylation (AcGEP). The physicochemical properties (such as solubility and substitution degree) and structural information (such as molecular weight Mw and radius of gyration Rg) of GEP derivatives were determined by Fourier transformed infrared (FTIR) spectroscopy and asymmetrical flow field-flow fractionation (AF4) coupled online with multiangle light scattering (MALS) and differential refractive index (dRI) detectors (AF4-MALS-dRI). The effects of the structural modification of GEP on the proliferation, apoptosis, and cell cycle of MCF-7 cell were studied systematically. The ability of MCF-7 cell for the uptake of GEP was studied by laser scanning confocal microscopy (LSCM). The results suggested that the solubility and anti-breast cancer activity of GEP were enhanced and the average Rg and Mw of GEP decreased after chemical modification. The AF4-MALS-dRI results showed that the chemical modification process simultaneously caused the degradation and aggregation of GEPs. The LSCM results revealed that more SGEP can enter the MCF-7 cell interior compared with AcGEP. The results indicated that the structure of AcGEP could play a dominating role in antitumor activity. The data obtained in this work can be used as a starting point for investigating the structure-bioactivity of GEPs.

Transcriptome analysis reveals the mechanism for blue-light-induced biosynthesis of delphinidin derivatives in harvested purple pepper fruit.

Anthocyanins are the main pigments that affect the color and quality of purple-fruited sweet pepper (Capsicum annuum). Our previous study indicated that blue light can induce anthocyanin accumulation in purple pepper. In view of its underlying mechanism that is unclear, here, anthocyanin content was determined, and transcriptome analysis was performed on pepper fruits harvested from different light treatments. As a result, among the identified anthocyanin metabolites, the levels of delphinidin (Dp) glycosides, including Dp-3-O-rhamnoside, Dp-3-O-rutinoside, and Dp-3-O-glucoside, were highly accumulated in blue-light-treated fruit, which are mainly responsible for the appearance color of purple pepper. Correlation between anthocyanin content and transcriptomic analysis indicated a total of 1,619 upregulated genes were found, of which six structural and 12 transcription factor (TF) genes were involved in the anthocyanin biosynthetic pathway. Structural gene, for instance, CaUFGT as well as TFs such as CaMYC2-like and CaERF113, which were highly expressed under blue light and presented similar expression patterns consistent with Dp glycoside accumulation, may be candidate genes for anthocyanin synthesis in response to blue-light signal.

[Separation and characterization of Gastrodia elata polysaccharides based on asymmetrical flow field-flow fractionation: steric transition phenomenon].

Asymmetrical flow field-flow fractionation (AF4), a gentle tool for the separation and characterization of particles and macromolecules, has attracted increased interest in recent years owing to its broad dynamic size range and utilization of "open channel" voids in the packing or stationary phase. A steric transition phenomenon in which the sample elution mode change from the normal mode to the steric/hyperlayer mode occurs. Accurate characterization by AF4 requires the absence of steric transition, particularly when the sample has a broad size distribution, because the effect of the combination of different modes is difficult to interpret. In this study, the relative molecular mass (M), radius of gyration (Rg), and conformation of Gastrodia elata polysaccharides (GEPs) were characterized using AF4 coupled with online multi-angle light scattering (MALS) and differential refractive index (dRI) detection (AF4-MALS-dRI). Steric transition was observed during GEP separation by AF4 owing to the broad size distribution of the molecules. This phenomenon would result in the inaccurate characterization of the GEPs in terms of M and Rg because two GEP groups of different sizes may elute together. In this study, the effects of constant and exponentially decaying cross-flow rates, sample mass concentration, and spacer thickness on steric transition were systematically investigated. The results indicated that a high GEP mass concentration (i. e., 0.75 mg/mL) can lead to steric transition. The spacer thickness affected the resolution and retention time of the GEPs and changed the steric transition point (di). An exponentially decaying cross-flow rate not only adjusted the di of the polydisperse GEP samples but also improved the GEP resolution and shortened the analysis time. The influence of steric transition was solved under the following operating conditions: injected GEP mass concentration=0.5 mg/mL; injection volume=50 µL; spacer thickness=350 µm; detector flow rate=1.0 mL/min; and cross-flow rate exponentially decayed from 0.2 to 0.05 mL/min with a half-life of 2 min. Moreover, the influence of GEP origins and ultrasound treatment time on the M and Rg distributions and conformation of GEPs were investigated under the optimized operating conditions. The results showed that the M and Rg distributions of Yunnan and Sichuan GEPs decreased with increasing ultrasound time. When the ultrasound treatment time was 15 min, the Yunnan GEPs had a loosely hyperbranched chain conformation, whereas the Sichuan GEPs had a spherical conformation. When the ultrasound treatment time was increased to 30 or 60 min, the GEPs from both Yunnan and Sichuan had a hyperbranched chain conformation, indicating that ultrasound treatment resulted in GEP degradation. Under the same extraction conditions, GEPs from Yunnan had larger M and Rg values than those from Sichuan. AF4-MALS-dRI showed good repeatability for the characterization of GEPs under the optimized operating conditions. The relative standard deviations of Rg and M were 0.5% and 1.7%, respectively. The data presented in this study can be used as a starting point for in-depth studies on the structural bioactivity of GEPs.

Study on effects of preparation method on the structure and antioxidant activity of protein-Tremella fuciformis polysaccharide complexes by asymmetrical flow field-flow fractionation.

Oxidation is an essential biological process for human life. In this study, low density lipoprotein-Tremella fuciformis polysaccharide (LDL-TFP) complexes were prepared by electrostatic and covalent methods. The effects of preparation method on the structure and antioxidant activity of LDL-TFP complexes were investigated by asymmetrical flow field-flow fractionation (AF4) coupled with ultraviolet-visible (UV/Vis), multiangle light scattering (MALS), and differential refractive index (dRI) detectors. The results showed that the electrostatic LDL-TFP complexes had a spherical structure, while the covalent LDL-TFP complexes had a rod-like structure as indicated by the ratio of Rg (radius of gyration) to Rh (hydrodynamic radius). Moreover, the results revealed that the antioxidant activity of the LDL-TFP complexes on the HepG2 could be related to the structure of LDL-TFP complexes. The antioxidant activity of LDL-TFP complexes formed by LDL modified with phospholipase A2 was further enhanced. This study would help expand the application of TFP.

Towards a better understanding of the relationships between the structure and antitumor activity of Gastrodia elata polysaccharides by asymmetrical flow field-flow fractionation.

To better understand the structure-function relationship of Gastrodia elata polysaccharides (PGEs), PGEs were extracted by ultrasound-assisted extraction method and the effects of extraction time on the structure and conformation of PGEs were evaluated by asymmetrical flow field-flow fractionation (AF4) coupled online with multiangle light scattering (MALS) and differential refractive index (dRI) detectors (AF4-MALS-dRI). Besides separation, AF4-MALS-dRI can provide more information about PGEs, such as size and molecular weight (Mw) distributions, apparent density, and conformation. The effects of PGEs on the proliferation, apoptosis, and cell cycle of MCF-7 cells were investigated. The cell activity assay indicated that the PGEs can inhibit the growth of MCF-7 cells by inducing late apoptosis. The results indicated that PGEs with a spherical conformation and compact structure seem to be beneficial to inducing MCF-7 cells late apoptosis. Moreover, results demonstrated that the information obtained by AF4-MALS-dRI is valuable for better understanding of the relationship of structure-activity of PGEs.

Applications of asymmetrical flow field-flow fractionation for separation and characterization of polysaccharides: A review.

Polysaccharides are the most abundant natural biopolymers on the earth and are widely used in food, medicine, materials, cosmetics, and other fields. The physicochemical properties of polysaccharides such as particle size and molecular weight often affect their practical applications. In recent years, asymmetrical flow field-flow fractionation (AF4) has been widely used in the separation and characterization of polysaccharides because it has no stationary phases or packing materials, which reduces the risk of shear degradation of polysaccharides. In this review, the principle of AF4 was introduced briefly. The operation conditions of AF4 for the analysis of polysaccharides were discussed. The applications of AF4 for the separation and characterization of polysaccharides from different sources (plants, animals, and microorganisms) over the last decade were critically reviewed.

Medical image enhancement in F-shift transformation domain.

Image enhancement technology plays an important role in the diagnosis and treatment of medical diseases. In this paper, we propose a method to automatically enhance medical images. The proposed method could be used to support clinical medical diagnosis, adjuvant therapy and curative effect diagnosis. This scheme uses contrast limited adaptive histogram equalization (CLAHE) method in F-shift transformation domain. Firstly, we adjust the overall brightness of the underexposed or overexposed image. Secondly, we perform CLAHE to enhance the low-frequency components obtained by one-level two-dimensional F-shift transformation (TDFS) on the adjusted images. At this stage, most of the coefficients in the high-frequency component can be changed to zero through properly setting the error bound. We then use inverse transformation to reconstruct image which is further enhanced with CLAHE. Compared to previous work, this approach takes into account not only the image enhancement, but also the data compression. Experimental results and comparison with state-of-the-art methods show that our proposed method has a better enhancement performance. Moreover, it has a certain data compression ability.

Generation of a long-acting fusion inhibitor against HIV-1.

AIDS has evolved from a fatal infectious disease to a manageable chronic disease under the treatment of anti-AIDS medications. HIV fusion inhibitors with high activity, low side effects and strong selectivity are promising drugs against HIV. Only one fusion inhibitor is currently approved, thereby highly active long-acting fusion inhibitors need to be developed for long-term AIDS treatment. Here, we synthesised MT-SC22EK (a small HIV fusion inhibitor) derivatives containing 1-2 staples to improve its stability. Antiviral activity studies showed that MT-SC22EK-2 with two staples exhibited potent inhibitory activity against HIV-1 standard strains and Chinese epidemic strains, and at the same time, MT-SC22EK-2 presented strong anti-T20 resistance. Surprisingly, MT-SC22EK-2 possessed excellent protease stability with a half-life of 3665 min. MT-SC22EK-2 is a potential HIV fusion inhibitor considered as a long-acting anti-HIV drug candidate.