Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related mortality worldwide, with systemic therapies offering only limited benefit. Sorafenib, the first-line multi-kinase inhibitor, is constrained by poor pharmacokinetics, dose-limiting toxicities, and the rapid emergence of resistance. Nanotechnology-based drug delivery systems (NDDS) provide promising strategies to address these limitations. This review summarizes recent NDDS innovations that enhance sorafenib’s therapeutic efficacy in HCC. Major nanocarrier classes, including lipid-based, polymeric, inorganic, and biogenic platforms, are discussed together with modifications for passive or active tumor targeting, such as asialoglycoprotein receptor-mediated approaches. Special attention is given to stimuli-responsive systems that exploit tumor microenvironmental cues to achieve localized drug release. Co-delivery strategies that combine sorafenib with chemotherapeutics, RNA interference agents, or immune modulators are also highlighted for their capacity to amplify antitumor activity and overcome resistance. Artificial intelligence (AI) is emerging as a tool to support the rational design and personalization of nanomedicines. While encouraging preclinical evidence demonstrates improved pharmacokinetics, tumor accumulation, and efficacy, challenges in scalable production, biosafety, and regulatory approval remain. Collectively, these developments emphasize the potential of sorafenib nanomedicines while underscoring the need to resolve key barriers for clinical translation.

Bletilla striata polysaccharide (BSP), a naturally derived glucomannan, has attracted increasing interest due to its excellent biocompatibility and diverse bioactivities. In this study, we investigated the potential of BSP as a photoprotective agent against ultraviolet A (UVA)-induced skin damage. Structural analysis confirmed that BSP is a linear β-(1 → 4)-glucomannan with a molecular weight of approximately 30.5 kDa, primarily composed of mannose and glucose units. In vitro, BSP treatment significantly improved the viability of UVA-irradiated keratinocytes. In vivo, BSP administration in a mouse model of UVA-induced photodamage effectively attenuated epidermal thickening, increased water content, and improved histological integrity. Mechanistically, BSP restored antioxidant capacity by activating the Nrf2/HO-1 signaling pathway and suppressed ferroptosis through upregulation of GPX4 and FTH1, reduction of lipid peroxidation and mitigation of iron accumulation. Multi-omics and immunofluorescence analyses further revealed cell type-specific responses and the central role of Nrf2 activation in mediating BSP's protective effects. Collectively, these findings highlight BSP as a promising natural polysaccharide for preventing UVA-induced oxidative skin damage and advancing the development of functional biomaterials for photoprotection.

Ultraviolet (UV) radiation is a primary factor contributing to photoaging, a form of premature skin aging characterized by the appearance of wrinkles, fine lines, uneven pigmentation, and reduced skin elasticity. Plantderived polysaccharides exhibit notable antioxidant and anti-inflammatory properties, making them promising candidates for the management of skin photoaging. Nevertheless, the hydrophilic nature and large molecular size of polysaccharides make them less effective for topical application. This study aimed to develop a method to increase polysaccharide transdermal absorption, using Crataegus pinnatifida polysaccharide (CPP) as a model. Acetylation was employed to modify the CPP, yielding three derivatives with varying degrees of substitution (DS): 0.16 (Ace-CPP1), 0.43 (Ace-CPP2), and 0.56 (Ace-CPP3). The in vitro antioxidant activity increased with increasing degree of substitution. A nanoemulgel formulation was developed, achieving approximately 72 % permeation of the native CPP. Furthermore, the acetylated CPP derivatives demonstrated enhanced permeation, exceeding 92 % within 4 h. In vivo studies revealed that the Ace-CPP3-based nanoemulgel significantly outperformed the native CPP in alleviating UVB-induced photoaging. This was evidenced by reduced oxidative stress, suppression of tissue inflammation, and promotion of collagen deposition. These findings underscore the potential of nanoemulgel formulation of acetylated CPP derivatives to advance applications in dermatology and cosmeceuticals.

The fungal cell wall provides the cell with enough strength to withstand turgor pressure and keeps adequate plasticity to extend the cell wall size under turgor pressure for cell growth. The cell walls of apical growing hyphae and budding growth yeast have been studied in detailed which share common components of chitin and β-1,3-glucan in their scaffold structures while other polysaccharide components vary on species. In contrast, the cell walls of elongating growth mushroom stipe remains poorly studied. This study explored that in addition to chitin, β-1,3-glucan with β-1,6-linkage branches, and β-1,6-glucan with β-1,3-linkage branches, the scaffold structure of C. cinerea cell wall also incorporated β-1,4-glucan, a component not previously reported in fungal cell walls. After converting chitin to chitosan, we identified three distinct forms of chitosan (chitin). The first was a free chitosan-β-glucan complex, in which chitosan was covalently linked to low molecular weight β-1,6-, β-1,3-, or β-1,4-glucan, which could be extracted using 10 % acetic acid. The second form consisted of chitosan-β-glucan complexes covalently linked to the β-1,4-glucan matrix polysaccharide via β-1,6-glucan, β-1,3-glucan, or β-1,4-glucan, which could be released from the cell wall by hydrolases hydrolysis. The third form involved the release of insoluble chitosan as chitooligosaccharides by chitosanase.

Ovomucin (OVM) is a key protein in egg white responsible for gel formation, with high glycosylation. To understand the association of glycosylation with OVM gelation capacity and structure stability, the effects and mechanisms of different glycan chains and monosaccharides on OVM gel properties were systematically investigated in this study. The results showed that removal of O-glycan chains rendered OVM completely incapable of gelation, whereas N-glycan chain removal reduced the cross-linking density of gel networks. Modification of OVM with neutral monosaccharides significantly enhanced the mechanical properties of gel, mainly attributed to the enhancement of disulfide and hydrogen bonds. The moderate presence of sialic acid (SA) facilitated the formation of OVM gels by regulating the molecular spacing via electrostatic forces. However, excess SA destabilized the gel network. In addition, the glycosylation sites and glycan chain composition of OVM were analyzed by LC-MS/MS, and the protective effect of OVM glycan chains to its own molecular structure under strong alkaline environment was revealed. Computational biology further validated the enhancement of intermolecular interactions by glycosylation. This study proved the crucial regulatory role of OVM glycosylation structure on its gel properties, providing a new insight into the strategy of modulating functional properties of proteins through glycosylation modifications.

Wound healing is a highly orchestrated, multiphase process that involves various cell types and molecular pathways. Recent advances in single-cell transcriptomics and machine learning have provided unprecedented insights into the complexity of this process, enabling the identification of novel cellular subpopulations and molecular mechanisms underlying tissue repair. In particular, single-cell RNA sequencing (scRNA-seq) has revealed significant cellular heterogeneity, especially within fibroblast populations, and has provided valuable information on immune cell dynamics during healing. Machine learning algorithms have enhanced data analysis by improving cell clustering, dimensionality reduction, and trajectory inference, leading to a better understanding of wound healing at the single-cell level. This review synthesizes the latest findings on the application of scRNA-seq and machine learning in wound healing research, with a focus on fibroblast diversity, immune responses, and spatial organization of cells. The integration of these technologies has the potential to revolutionize therapeutic strategies for chronic wounds, fibrosis, and tissue regeneration, offering new opportunities for precision medicine. By combining computational approaches with biological insights, this review highlights the transformative impact of scRNA-seq and machine learning on wound healing research.

In this study, we prepared four derivatives of fucosylated chondroitin sulfate (FCS): full-length FCS (flFCS) from Holothuria leucospilota, low molecular weight FCS (lmFCS) derived from flFCS, and their de-branched counterparts, de-branched flFCS (d-flFCS) and de-branched lmFCS (d-lmFCS) via controlled acid treatment. Following structural verification using various analytical techniques, we applied targeted metabolomics to examine the impact of FCS on nutritional efficacy and its structure-activity relationship. Analysis of 225 plasma and feces samples from 75 mice revealed a positive correlation between metabolomic shifts and increased weight gain, underscoring FCS's potential to enhance nutrient absorption and promote growth. The observed linear relationship between the levels of short-chain fatty acids in plasma and feces suggests that FCS may facilitate catabolic activities in the gastrointestinal tract. The comparative study of different FCS derivatives on mouse growth and metabolic homeostasis regulation led to the conclusion that FCS exhibits greater biological activity with a higher degree of branching and larger molecular weight.

Heparin is the most extensively used anticoagulant in clinical practice. It is a highly sulfated, linear polysaccharide composed of repeating disaccharide units. As a member of the glycosaminoglycan (GAG) family, heparin's complex structure features significant molecular weight variability, diverse sugar residues, and variable sulfation patterns. Low molecular weight heparins (LMWHs), produced through chemical or enzymatic depolymerization, are distinguished by their reduced molecular weight and offer therapeutic advantages, including lower bleeding risks, reduced immunogenicity, and higher bioavailability following subcutaneous administration. The structural intricacy of heparin-based drugs presents major challenges for quality control, clinical safety, process optimization, and therapeutic expansion. Advanced analytical methods, particularly LC and MS, remain at the forefront of efforts to elucidate the detailed structures of these drugs. This review highlights recent progress in chromatographic and MS-based analysis techniques for heparin and its derivatives, including the application of computational algorithms for structural elucidation. The focus is on the analytical methodologies, their innovations, and limitations, while also exploring how machine learning and bioinformatics tools are shaping the future of heparin quality control and therapeutic application. This comprehensive review provides a reference point for researchers engaged in the structural analysis of heparin-based drugs and offers insights into the future development of novel analytical strategies for improving the safety and efficacy of these critical anticoagulants.

Carbohydrates are essential biomolecules that play a vital role in various biological processes across humans, plants, and bacteria. Despite their ubiquity, the structural elucidation of carbohydrates, particularly oligo- and polysaccharides, remains a significant challenge due to their complex and heterogeneous nature. The high-performance anion exchange chromatography (HPAEC) or called ion chromatography (IC) coupled with pulsed amperometric detection (PAD) has emerged as a powerful tool for highly effective separation and highly specific detection of glycans. The introduction of mass spectrometry (MS) into HPAEC-PAD systems has further advanced glycan analysis by enabling detailed structural elucidation, including branching, linkage patterns, and sequence determination. The use of suppressor technology allows for the coupling of HPAEC with MS by converting non-volatile salts in the mobile phase into volatile ones. This review highlights the current advancements in HPAEC-PAD/MS for oligo- and polysaccharide structural analysis, discussing the strengths and limitations of different suppressor systems, the role of MS in glycan analysis, and the emerging applications of this technology in the field of glycomics. With continued innovation, HPAEC-PAD/MS is poised to become an essential tool for the detailed characterization of polysaccharides, supporting advancements in pharmaceutical, biomedical, and biotechnological research.

The swollen culm (also known as Jiaobai) of Zizania latifolia is formed by the smut Ustilago esculenta invades the Z. latifolia. The new tissue formed due to the symbiotic relationship has entices the attention of researchers to study its polysaccharide structure along with biological evaluation. Five fractions of polysaccharides were obtained owing to hot water extraction, alcoholic precipitation, and chromatographic purification. Bioactivity assays showed that ZLPs have good antioxidant, hypoglycemic activities and protective activity against oxidative damage. The ZLP-1 and ZLP-2 were determined to be neutral polysaccharides with high purity, exhibiting propitious bioactivity, consequently they were subjected to indispensable structural characterization. These results showed that ZLP-1 has molecular weight (Mw) of 103 kDa and glucose (Glc) (76.68 %) as the primary monosaccharide; the ZLP-2 has Mw of 122 kDa and galactose (Gal) (41.04 %) and arabinose (Ara) (27.12 %). Structural elucidation by methylation and nuclear magnetic resonance (NMR) analysis suggested ZLP-1 is a glucan, with →3)-β-Glcp-(1→3)-β-Glcp-(1→4)-β-Glcp-(1→4)-β-Glcp-(1→3,6)-β-Galp-(1→3)-β-Glcp-(1→ as the mainchain and the terminal Araf and Glcp; the ZLP-2 is a Galactoxylan, with →3,4)-β-xylp-(1→3)-β-Galp-(1→3,6)-β-Galp-(1→3,6)-β-Galp-(1→ as the mainchain and the terminal Araf and Glcp. The structural arrangements provide a chemical basis for understanding the nutritional and pharmacological activities of polysaccharides from Zizania latifolia.

Sulodexide, a heparinoid medicine, is wildly used in clinic for prophylaxis and treatment of thromboembolic diseases and diabetic nephropathy. Despite its widespread use, the structure of Sulodexide remains poorly understood. It consists of various polysaccharides characterized by differing sugar compositions, linkages, and sulfonation patterns, yet they share common features such as strong hydrophilicity, high native charges, and considerable polydispersity, posing significant challenges for conventional chromatographic and online mass spectrometry (MS) characterization. In this work, a novel analytical method combining multiple-heart cut 2D-LC and in-source acid-induced dissociation (inAID) MS was developed. Three polysaccharides in Sulodexide were separated by high efficient strong-anion-exchange chromatography, followed by desalting with the second dimensional size-exclusion chromatography before MS. A novel MS strategy employing inAID technique was utilized for online analysis, leading to the initial identification of Sulodexide polysaccharide components. The results were validated through disaccharide composition analysis of those three polysaccharide components after offline preparation. This advanced strategy, merging various techniques, enable a comprehensive structural elucidation of such complex drugs and provides a viable tool for potential routine analysis of complex biomolecules.

Heparin is the most widely used anticoagulant in clinical practice, with enoxaparin being one of the most important low molecular weight heparins (LMWHs). In this study, an antithrombin III (ATIII) affinity column was used. Enoxaparin and its oligosaccharides of varying sizes, prepared using preparative size exclusion chromatography (SEC), were fractionated through the ATIII affinity column. The different affinity fractions from each oligosaccharide size were profiled using strong anion exchange (SAX) chromatography. Each peak was automatically transferred to an SEC column for desalting prior to mass spectrometry (MS) analysis, which enabled structural identification using a multiple heart-cut (MHC) 2D LC-MS system (SAX-SEC-MS). The high-affinity fraction from enoxaparin was further analyzed using the MHC 2D LC system (SEC-SAX). SAX profiles of the high-affinity oligosaccharides, prepared by both size and affinity fractionation, were consistent with those obtained by direct SEC-SAX analysis. The possible sequences of several high-affinity hexasaccharides and the domain compositions of high-affinity octa- and decasaccharides in enoxaparin were further elucidated by disaccharide analysis after manual collection of the oligosaccharides. This work advances the understanding of enoxaparin's structural features and offers a potential approach to improve the quality of enoxaparin, as well as to identify key structural motifs in heparin/LMWHs that contribute to protein binding.

The dried root of Pueraria mirifica (P. mirifica) is an edible foodstuff widely used in Asian countries. P. mirifica is known for its high starch content. The isolation of polysaccharides from high-starch plant parts is challenging due to the interference of starch. Therefore, this study aimed to develop a technique for isolating and investigating the structure and activity of non-glucan polysaccharides from P. mirifica (PMP). An effective starch removal process was developed using α-amylase hydrolysis and thorough membrane dialysis. Four non-glucan polysaccharides were isolated, and PMP-2 was subjected to structural elucidation. The results indicated that PMP-2 has a molecular weight of 124.4 kDa and that arabinose and galactose are the main components, accounting for 27.8 % and 58.5 %, respectively. Methylation and NMR analysis suggested that PMP-2 is an Arabinogalactan composed of 1,6-linked Galp and 1,4-linked Galp as the main chain, with arabinan and rhamnose as side chains. Furthermore, PMP-C and PMP-2 exhibited concentration-dependent antioxidant activities against DPPH, ABTS, and hydroxyl radicals and certain immunomodulatory activities related to the release of NO, TNF-α and IL-6. These findings suggest that PMP-2 has potential therapeutically active ingredient in functional foods. The developed method successfully removed starch and isolated non-glucan polysaccharides from the high-starch content plant P. mirifica and can be applied to other high-starch plants.

Metabolic alterations within mitochondria, encompassing processes such as autophagy and energy metabolism, play a pivotal role in facilitating the swift proliferation, invasion, and metastasis of cancer cells. Despite this, there is a scarcity of currently available medications with proven anticancer efficacy through the modulation of mitochondrial dysfunction in a clinical setting. Here, we introduce the structural characteristics of RN0D, a galactoglucan isolated and purified from Panax notoginseng flowers, mainly composed of β-1,4-galactan and β-1,3/1,6-glucan. RN0D demonstrates the capacity to induce mitochondrial impairment in cancer cells, leading to the accumulation of reactive oxygen species, initiation of mitophagy, and reduction in both mitochondrial number and size. This sequence of events ultimately results in the inhibition of mitochondrial and glycolytic bioenergetics, culminating in the demise of cancer cells due to adenosine triphosphate (ATP) deprivation. Notably, the observed bioactivity is attributed to RN0D's direct targeting of Galectin-3, as affirmed by surface plasmon resonance studies. Furthermore, RN0D is identified as an activator of the PTEN-induced kinase 1 (PINK1)/Parkin pathway, ultimately instigating cytotoxic mitophagy in tumor cells. This comprehensive study substantiates the rationale for advancing RN0D as a potentially efficacious anticancer therapeutic.

In this work, a synthetical glycan fingerprinting strategy using a multiple heart-cut two-dimensional liquid chromatography system linked to mass spectrometry (MHC 2D LC-MS) was developed to analyze enoxaparin, a widely used low molecular weight heparin (LMWH). Glycans from an enoxaparin standard were prepared offline based on size, and the derived tetra-, hexa-, octa- and decasaccharides were profiled using a qualitative analytical platform. Strong anion exchange chromatography (SAX) was employed as the first-dimensional chromatography (1D) to separate glycans of the same size but with different charges or sequences, while size exclusion chromatography (SEC) was used in the second dimension (2D) for desalting before MS analysis. The retention times (RTs), accurate masses and structural compositions of the glycans were fully characterized. Real samples were analyzed using the same platform but with 1D and 2D exchanged: SEC was used to separate enoxaparin glycans by size, followed by SAX using the same parameters as in the qualitative platform to separate glycans by charge or sequence. Glycans in real samples were identified by matching their RTs to those assigned in the qualitative analysis and semi-quantitated after normalization of peaks in each SAX chromatogram of glycans of different sizes. The analyses were performed automatically and robustly using this synthetical platform, enabling the fingerprinting and differentiation of enoxaparins from various sources. This platform could serve as a powerful tool for structural analysis, quality control, and heparin-related drug development.

In this study, 37 derivatives of phorbol esters were synthesized and their anti-HIV-1 activities evaluated, building upon our previous synthesis of 51 phorbol derivatives. 12-Para-electron-acceptor-trans-cinnamoyl-13-decanoyl phorbol derivatives stood out, demonstrating remarkable anti-HIV-1 activities and inhibitory effects on syncytia formation. These derivatives exhibited a higher safety index compared with the positive control drug. Among them, 12-(trans-4-fluorocinnamoyl)-13-decanoyl phorbol, designated as compound 3c, exhibited the most potent anti-HIV-1 activity (EC50 2.9 nmol·L−1, CC50/EC50 11 117.24) and significantly inhibited the formation of syncytium (EC50 7.0 nmol·L−1, CC50/EC50 4891.43). Moreover, compound 3c is hypothesized to act both as an HIV-1 entry inhibitor and as an HIV-1 reverse transcriptase inhibitor. Isothermal titration calorimetry and molecular docking studies indicated that compound 3c may also function as a natural activator of protein kinase C (PKC). Therefore, compound 3c emerges as a potential candidate for developing new anti-HIV drugs.

In this work, the process of forming PF4/heparin complex was described based on their sizes measured with PCS. PF4/heparin complexes formed when a small amount of heparin was added to PF4, and they grew to bigger particles when more heparin was added, but they started to decompose after a certain amount of heparin was added. The larger the molecular weight of heparin has, the less amount of heparin is required to form the complex with PF4. This process was verified by the results of the complex surface charge. This method was used to compare the immunogenicity among the different batches of nadroparin after its specificity and stability were validated.

Heparin-induced thrombocytopenia (HIT) is an immune complication of heparin therapy. Antibodies binding to complexes of platelet factor 4 (PF4) and heparin is the trigger of HIT. A method using size exclusion chromatography with multi-angle laser light scattering detector (SEC-MALS) was developed in this work. The soluble ultra-large complex (ULC) was separated from the small complex (SC) and their molecular weights (MWs) were firstly measured. The complexes of PF4 and three heparins with different MW, including unfractionated heparin (UFH), dalteparin (Daltep) and enoxaparin (Eno) were characterized using this method. The contents and the sizes of ULC increased gradually when heparins were added to PF4 to certain amounts. While, they reduced after more heparins were added. It is the first time to measure the MWs of the biggest ULC of PF4-heparins as millions of Dalton. at the proper ratios of PF4 to heparin (PHR). Meanwhile, those mixtures at those certain PHRs induced the higher expression of CD83 and CD14 markers on dendritic cells (DCs) suggesting that they had stronger immunogenicity and is critical for HIT.

Glycyrrhizae Radix et rhizome/licorice is a precious herb in traditional Chinese medicine (TCM). TCM's polysaccharides are medicinally active. But herbal polysaccharides pose some limitations for topical applications. Therefore, this study aimed to utilize licorice polysaccharide via mesoporous silica nanoparticles (MSN) for antiacne efficacy in topical delivery. The polysaccharide (GGP) was extracted with a 10 % NaOH solution. Chemical characterization suggested that GGP possesses an Mw of 267.9 kDa, comprised primarily of Glc (54.1 %) and Ara (19.12 %), and probably 1,4-linked Glc as a backbone. Then, MSN and amino-functionalized MSN were synthesized, GGP entrapped, and coated with polydopamine (PDA) to produce nanoparticle cargo. The resulted product exhibited 76 % entrapment efficiency and an in vitro release of 89 % at pH 5, which is usually an acneprone skin's pH. Moreover, it significantly increased Sebocytes' cellular uptake. GGP effectively acted as an antiacne agent and preserved its efficacy in synthesized nanoparticles. In vivo, the results showed that a 20 % gel of MSN-NH2-GGP@PDA could mediate an inflammatory response via inhibiting pro-inflammatory cytokines and regulating anti-inflammatory cytokines. The MSN-NH2-GGP@PDA inhibited TLR2-activated-MAPK and NF-kappa B pathway triggered by heat-killed P. acnes. In conclusion, fabricated MSN entrapped GGP for biomimetic anti-acne efficacy in topical application.

Gentiana dahurica Fisch. ( G. dahurica ) is one of the legitimate sources of Qinjiao in Traditional Chinese Medicine (TCM) and grows on high-altitude plateaus. Plants develop unique biochemical accumulations to resist plateau conditions, especially the strong UV irradiation. Thus, this study aimed to investigate the polysaccharide of G. dahurica (GDP), its structure and its activity against UVB irradiation. Four GDPs were isolated and two of them were subjected to structural elucidation. The results suggested that GDP-1 has 53.5 % Ara and 30.8 % GalA as its main monosaccharides, with a molecular weight (Mw) of 23 kDa; the GDP-2 has 33.9 % Ara and 48.5 % GalA, with a Mw of 82 kDa. Methylation and NMR spectroscopy analysis revealed that GDP-1 contains -> 5)- alpha-Ara f -(1 -> 5)- alpha-Ara f -(1 -> 3,5)- alpha-Ara f -(1 -> 3,4)- alpha-Gal p A-(6-OMe)-(1 -> as the main chain, the branches of GalA (with esterification), and the terminal Ara; the GDP-2 contains -> 4)- alpha-Gal p A-(1 -> 4)- alpha-Gal p A-(6-OMe)-(1 -> 5)- alpha-Ara f - (1 -> 3,5)- alpha-Ara f -(1 -> as the main chain, the branches of -> 5)- alpha-Ara f -(1 - 5)- alpha-Ara f , and the terminal GalA. Both GDP-1 and GDP-2 exhibited concentration-dependent antioxidant activity against DPPH, ABTS and hydroxyl radicals. Moreover, GDPs significantly attenuated the decreases in viability and proliferation of HaCaT cells after UVB irradiation. They can scavenge reactive oxygen species (ROS) and improve the activities of endogenous antioxidant enzymes, including superoxide dismutase (SOD) and glutathione peroxidase (GSH). The potential mechanism explored by flow cytometry assays of cell apoptosis and cell cycle distribution suggested that GDPs exert protective effects against UVB irradiation by reducing ROS and attenuating S phase cell arrest. In brief, the GDP-1 and GDP-2 are alpha-1,3- and alpha-1,4- arabinogalacturonan, respectively. The high content of Ara could be attributed to biochemical accumulation in resisting to the plateau environment and to prevent UVB irradiationrelated damage in cells. These findings provide insight into authentic medicinal herbs and the development of GDPs in the modern pharmaceutical and cosmetics industry.