Kiwi-derived extracellular vesicles for oral delivery of sorafenib.

Sorafenib is an oral treatment for hepatocellular carcinoma (HCC). However, poor water solubility, harsh gastrointestinal environment and off-target effects contribute to the low bioavailability of oral sorafenib. Plant-derived extracellular vesicles (PDEVs) are biological nanovesicles with various bioactive functions that offer significant advantages in the field of oral drug delivery: protection from degradation by gastrointestinal fluids; crossing the intestinal epithelial barrier; specific targeting; safety; and abundant yield. However, there are fewer studies applying PDEVs for anti-tumor drug delivery to extra-digestive tissues. In this study, kiwifruit-derived extracellular vesicles (KEVs) were isolated and purified from kiwifruit, and their natural hepatic accumulation properties were exploited for targeted delivery of sorafenib (KEVs-SFB). Evidence showed that encapsulation of KEVs reduced the leakage of sorafenib in the gastrointestinal environment and enhanced the ability to cross the intestinal epithelium; KEVs-SFB was able to achieve liver accumulation and was predominantly taken up by HepG2 cells; KEVs-SFB was effective in inhibiting 4T1 cell proliferation; in the orthotopic liver cancer model, oral administration of KEVs-SFB inhibited tumor growth and improved the side effects of SFB. This PDEVs-based oral drug delivery platform is important for improving oral bioavailability and reducing drug side effects.

Semi-solid enzymolysis enhanced the protective effects of fruiting body powders and polysaccharides of Herinaceus erinaceus on gastric mucosal injury.

This study demonstrated the effects of semi-solid enzymolysis on physicochemical properties of fruiting body powders and polysaccharides from Hericium erinaceus and protective effects on gastric mucosal injury. Semi-solid enzymolysis could reduce the particle size, change the microstructure of fruiting body powders, increase the contents of soluble polysaccharide (26.26-67.04 %) and uronic acid (16.97-31.12 %) and reduce the molecular weight of polysaccharides. The digestibility of fruiting body powder of H. erinaceus after semi-solid enzymolysis was increased by 31.4 %, compared with that of the fruiting body powder of H. erinaceus without enzymolysis. Semi-solid enzymolysis could enhance the protective effects of the fruiting body powders and polysaccharides on ethanol-induced human gastric mucosal epithelial cells (GES-1) cells, increase the production of superoxide dismutase (SOD, 0-37.33 %) and catalase (CAT, 2.47-18.46 %), and inhibit the production of malonaldehyde (MDA, 2.45-19.62 %), myeloperoxidase (MPO, 0-13.54 %), interleukin (IL-6, 4.39-24.62 %) and tumor necrosis factor-α (TNF-α, 5.97-12.25 %). Semi-solid enzymolysis could improve the inhibition rate of the fruiting body powder on gastric ulcer (32.70-46.26 %), inhibit oxidative stress and inflammation, and protect rats with acute gastric mucosal injury against the stimulation of ethanol on gastric mucosa. In conclusion, semi-solid enzymolysis may enhance the protective effects of the fruiting body powders and polysaccharides on gastric mucosal injury.

A cancer cell membrane coated nanoparticles-based gene delivery system for enhancing cancer therapy.

Gene therapy is a superior therapeutic means in cancer therapy. However, the instability of nucleic acid and the lack of suitable delivery carrier greatly restricts its further development and application. Herein, we coupled low molecular weight polyethyleneimine (LMW PEI) through disulfide bonds, then modified it with manganese dioxide (MnO2) nanosheets and nuclear localization signal peptide (NLS), as a p53 gene carrier, and finally coated it with B16F10 cell membrane to construct a novel gene-carrier system CM@MnO2-PEI-NLS-ss/p53 (M@MPNs/p53). Tumor cell membrane coating endows nanoparticles with homotypic targeting and immune escape capabilities, disulfide-crosslinked LMW-PEI has high transfection efficiency and low toxicity, and NLS peptides enhance nuclear delivery and improve p53 gene delivery efficiency; meanwhile, MnO2 nanosheets oxidize high intracellular concentration of glutathione (GSH), sensitizing p53 gene-mediated antitumor therapy. The results showed that the novel biofilm-camouflaged M@MPNs/p53 nanoparticles had a highly specific targeting effect on homologous cancer cells and could effectively inhibit tumor growth in vitro and in vivo. Besides, MnO2 loading improved p53-mediated tumor regression. This novel gene delivery platform is of great significance in improving gene delivery efficiency and enhancing anti-tumor therapy.

Advanced research on extracellular vesicles based oral drug delivery systems.

The oral route is the most convenient and simplest mode of administration. Nevertheless, orally administration of some commonly used therapeutic drugs, such as polypeptides, therapeutic proteins, small-molecule drugs, and nucleic acids, remains a major challenge due to the harsh gastrointestinal environment and the limited oral bioavailability. Extracellular vesicles (EVs) are diverse, nanoscale phospholipid vesicles that are actively released by cells and play crucial roles in intercellular communications. Some EVs have been shown to survive with the gastrointestinal tract (GIT) and can cross biological barriers. The potential of EVs to cross the GIT barrier makes them promising natural delivery carriers for orally administered drugs. Here, we introduce the uniqueness of EVs and their feasibility as oral drug delivery vehicles (ODDVs). Then we provide a general description of the different cellular EVs based oral drug delivery systems (ODDSs) currently under study and emphasize the contribution of endogenous features and multifunctional properties of EVs to the delivery performance. The current obstacles of moving EVs based ODDSs from bench to bedside are also discussed.

Phragmites rhizoma polysaccharide-based nanocarriers for synergistic treatment of ulcerative colitis.

The purpose of this study was to construct Phragmites rhizoma polysaccharide-based nano-drug delivery systems (PRP2-SeNPs-H/Aza-Lips) for synergistically alleviating ulcerative colitis and to investigate the important roles of Phragmites rhizoma polysaccharide-based nanocarriers in PRP2-SeNPs-H/Aza-Lips. Phragmites rhizoma polysaccharide (PRP2) was isolated and used for the preparation of Phragmites rhizoma polysaccharide selenium nanoparticles with low selenium content (PRP2-SeNPs-L) and high selenium content (PRP2-SeNPs-H). Based on the electrostatic attraction between PRP2-SeNPs-H and azathioprine liposomes (Aza-Lips), PRP2-SeNPs-H/Aza-Lips were constructed for precise delivery of the model drug azathioprine (Aza) to colon lesions. Results showed that PRP2 significantly alleviated the clinical symptoms and colon tissue damage and down-regulated the levels of inflammatory factors in serum and colon, demonstrating beneficial effects on mice with ulcerative colitis. PRP2-SeNPs-L had better relieving effects on ulcerative colitis. Phragmites rhizoma polysaccharide-based nanocarriers may protect azathioprine liposomes against gastrointestinal digestion, enhance the therapeutic effects on ulcerative colitis, and significantly reduce liver damage from azathioprine, which helps to improve the efficacy and toxicity of clinical drugs.

Isolation, Structural Analysis and Anti-Inflammatory Activity of a Polysaccharide from Ilex cornuta Fruits.

In the present study, a polysaccharide from Ilex cornuta fruits (LCFP-3) was obtained by hot water extraction, Diethyaminoethyl cellulose-52 (DEAE-52) chromatography column and Sephadex G-100 gel column purification. Its structural characteristics were further explored using high performance anion exchange chromatography (HPAEC), gas chromatography and mass spectrometry (GC/MS), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. Monosaccharide composition analysis revealed LCFP-3 contained mainly Galactose (31.92 %), Arabinose (25.87 %) and Galacturonic acid (23.35 %) while small percentage of Rhamnose, Glucose, Mannose and Xylose. Chemical composition analysis showed that the total sugar content of LCFP-3 was 90.31 % and the protein content was 0.246 %. Gel permeation chromatography (GPC) analysis showed that its average molecular weight was 41.199 kDa. Structural analysis showed that LCFP-3 may be composed of residues, T-α-Arap, T-α-Rhap, 1,3-α-Arap, 1,4-α-Arap, T-ß-Galp, 1,4-α-GalpA(OMe), 1,4-ß-Glcp, 1,3-ß-Galp, 1,3,6-ß-Manp, 1,6-ß-Galp, 1,3,4-ß-GalpA, 1,4,6-ß-Manp, 1,3,6-ß-Glcp, 1,2,3,4-α-Xylp. The anti-inflammatory activity of LCFP-3 was evaluated using lipopolysaccharide (LPS)-induced RAW246.7 macrophages. The results showed that 1-200 µg/mL LCFP-3 could dose-dependently protect against LPS-induced toxicity and 1 µg/mL LCFP-3 could significantly inhibit LPS-induced NO production. Therefore, LCFP-3 exerted an anti-inflammatory activity and has great potential as a functional ingredient.

Multifunctional metal complex-based gene delivery for tumour immune checkpoint blockade combination therapy.

Immune checkpoint blocking based on the PD-1/PD-L1 pathway has shown exciting results in various types of cancer. However, due to the off-target effect of PD-1/PD-L1 blocker, low tumour immunogenicity and tumour immunosuppressive microenvironment, a significant proportion of patients do not benefit from this treatment. Here, we constructed a novel multifunctional metal complex Fe/PEI-Tn by the coordination of polyethyleneimine (PEI) with Fe3+ and the modification of bifunctional peptides Tn containing the cell penetrating peptide (TAT) and nuclear localisation signal peptide (NLS), which was coated with hyaluronic acid (HA) to prolong the circulation time in vivo. Fe/PEI-Tn can condensate PD-L1 trap plasmid (pPD-L1 trap) and mediate PD-L1 trap protein expression in tumour tissues in situ, thus blocking the PD-1/PD-L1 pathway. Besides, Fe/PEI-Tn metal complex itself can act as an immune adjuvant to activate macrophages, reverse the phenotype of pro-tumour M2-type macrophages, and promote anti-tumour immunity. Meanwhile, Fe/PEI-Tn treatment can induce damage in tumour cells and release tumour-specific antigens into tumour microenvironment, thus stimulating anti-tumour immune response. Studies showed that HA/Fe/PEI-Tn/pPD-L1 trap complexes could promote the immune activation of tumour tissues and effectively delay tumour growth. This strategy provides a new direction for tumour combination therapy based on PD-1/PD-L1 blockade.

Therapeutic strategies to overcome cisplatin resistance in ovarian cancer.

Ovarian cancer (OC) is one of the most common gynecologic tumors worldwide and one with the highest mortality. Cisplatin (DDP) is the first platinum-based complex approved by the Food and Drug Administration (FDA) to treat patients with OC. Despite a good initial response rate, most patients receiving DDP treatment will ultimately develop resistance via various complicated mechanisms, leading to therapeutic failure and increased mortality. Multiple resistance pathways have been identified as potentially key areas of intervention. In this review, chemotherapeutic drugs and phytochemicals developed to overcome cisplatin-resistance ovarian cancer (CROC) were discussed. Targeted inhibition or specific drugs are effective against the DDP-resistance phenotype by inhibiting resistance or increasing cytotoxic efficacy. Phytochemicals as chemosensitizers offer novel treatment strategies for CROC patients by reducing chemoresistance and increasing drug efficacy. Due to the complexity of the DDP-resistance mechanism, the treatment of OC needs to improve specificity and effectiveness, and multi-path cooperative therapy is undoubtedly one of the best options. We discuss extensively the role of combination therapy in reversing DDP-resistance in OC and the significance of using a nanoparticle delivery system in this context. Suggestions for potential therapeutic strategies for CROC treatment will help discover more effective and specific regimens to overcome DDP-resistance.

Structure Analysis and Anti-Inflammatory Effect of a Polysaccharide from Hippocampus mohnikei.

The purpose of this study was to analyze the structure of a polysaccharide (HMP-1) from Hippocampus mohnikei, and to explore its anti-inflammatory effect. HMP-1 was obtained from Hippocampus mohnikei by ethanol sedimentation and secondary column chromatography purification. Its structural characteristics were analyzed by gel permeation chromatography (GPC), Fourier transform infrared (FT-IR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy and scanning electron micrograph (SEM). Results showed its molecular weight (Mw) was 7296 Da, and it mainly consisted of six residues: 1,3-ß-Glcp, 1,4-α-Manp, 1,4-α-GalpA, 1,4-ß-GlcpA2S, 1,4-α-Galp3S, and 1,4-ß-GlcNAc. HMP-1 could protect RAW246.7 cells from the cytotoxic effect induced by LPS. HMP-1 also could reduce the levels of nitric oxide and reactive oxygen species produced by LPS stimulation, suggesting that HMP-1 has anti-inflammatory activities within a certain concentration range.

Reversing tumor immunosuppressive microenvironment via targeting codelivery of CpG ODNs/PD-L1 peptide antagonists to enhance the immune checkpoint blockade-based anti-tumor effect.

In order to reverse tumor immunosuppressive microenvironment and improve antitumor immune effect based on immune checkpoint blocking, a mannose-modified liposome-based CpG ODNs and PD-L1 antagonistic peptides (P) co-delivery system (HA/M-Lipo CpG-P) was constructed, in which hyaluronic acid (HA) coating was supposed to improve the systemic circulation stability and thereby promote its accumulation in tumor tissues. When the HA/M-Lipo CpG-P complexes enter the tumor tissues, HA will be hydrolyzed under the action of hyaluronidase, exposing P peptides. Then, P peptides linked by octapeptides that can be cleaved by matrix metalloproteinases (MMPs) are released into tumor tissues under the action of MMPs, exerting a blocking effect in the PD-1/PD-L1 pathway. The M-Lipo CpG complexes can recognize macrophage surface mannose receptors through its surface modified mannose molecules, and promote the intracellular delivery of CpG ODNs, thereby activating macrophages. The results showed that HA/M-Lipo CpG-P complexes successfully reversed M2-type macrophages in tumor microenvironment (TME) to M1, thereby activating anti-tumor related immune cells and inhibiting tumor growth. Moreover, the HA/M-Lipo CpG-P complexes showed a better tumor inhibitory effect than the HA/M-Lipo CpG or the HA/M-Lipo-P (monotherapy) treatment groups. Overall, HA/M-Lipo CpG-P complexes provide a promising co-delivery strategy for targeting tumors to improve the antitumor effect based on immune checkpoint blockade.

Cereus sinensis Polysaccharide Alleviates Antibiotic-Associated Diarrhea Based on Modulating the Gut Microbiota in C57BL/6 Mice.

The present study investigated whether the purified polysaccharide from Cereus sinensis (CSP-1) had beneficial effects on mice with antibiotic-associated diarrhea (AAD). The effects of CSP-1 on gut microbiota were evaluated by 16S rRNA high-throughput sequencing. Results showed that CSP-1 increased the diversity and richness of gut microbiota. CSP-1 enriched Phasecolarctobacterium, Bifidobacterium and reduced the abundance of Parabacteroides, Sutterella, Coprobacillus to near normal levels, modifying the gut microbial community. Microbial metabolites were further analyzed by gas chromatography-mass spectrometry (GC-MS). Results indicated CSP-1 promoted the production of various short-chain fatty acids (SCFAs) and significantly improved intestinal microflora dysfunction in AAD mice. In addition, enzyme linked immunosorbent assay and hematoxylin-eosin staining were used to assess the effects of CSP-1 on cytokine levels and intestinal tissue in AAD mice. Results demonstrated that CSP-1 inhibited the secretion of interleukin-2 (IL-2), interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) and improved the intestinal barrier. Correspondingly, the daily records also showed that CSP-1 promoted recovery of diarrhea status score, water intake and body weight in mice with AAD. In short, CSP-1 helped alleviate AAD by regulating the inflammatory cytokines, altering the composition and richness of intestinal flora, promoting the production of SCFAs, improving the intestinal barrier as well as reversing the dysregulated microbiota function.

Colon-targeted drug delivery of polysaccharide-based nanocarriers for synergistic treatment of inflammatory bowel disease: A review.

Drugs such as immunosuppressants and glucocorticoids used for the treatment of inflammatory bowel disease (IBD) have certain troubling side effects. Polysaccharide-based nanocarriers with high safety and bioavailability are often used in the construction of colon-targeted drug nanodelivery systems (DNSs). It can help the drug resist the harsh environment of gastrointestinal tract, improve stability and concentrate on the intestinal inflammation regions as much as possible, which effectively reduces drug side effects and enhances its bioavailability. Certain polysaccharides, as prebiotics, can not only endow DNSs with the ability to target the colon based on enzyme responsive properties, but also cooperate with drugs to alleviate IBD due to its good anti-inflammatory activity and intestinal microecological regulation. The changes in the gastrointestinal environment of patients with IBD, the colon-targeted drug delivery process of polysaccharide-based nanocarriers and its synergistic treatment mechanism for IBD were reviewed. Polysaccharides used in polysaccharide-based nanocarriers for IBD were summarized.

Seed Oil Quality and Cultivation of Sambucus williamsii Hance as a New Oil Crop.

Natural edible oil derived from wild non-cultivated oil crops contributed to human daily nutritional diversity and disease prevention. It was important to investigate the nutritional value of these oils and the feasibility of crop cultivation. The present study focused on the assessment of seed oil quality of Sambucus williamsii Hance (SWH) and its molecular breeding. Wild SWH seed oil was extracted by supercritical CO2 technology and the composition of the oil was determined by using gas chromatography mass spectrometry (GC-MS) analysis. The oil content of SWH seeds reaches around 40%. Its seed oil was found to be rich in unsaturated fatty acids, such as 24.24% of linolenic acid and 50.56% of linoleic acid, and vitamin E (25.92 mg kg-1). The cytotoxicity and heavy metal analysis showed SWH seed oil was safe for consumption. In addition, the SWH strains with excellent characteristics were screened out for cultivation according to genetic diversity and morphological analysis. Amplified fragment length polymorphism (AFLP) markers were used to evaluate the genetic diversity of 28 accessions of wild SWH seeds and 5 accessions were selected to cultivate. Among them, two strains of SWH (sample 3 and 6) with high yielding (275.7 and 266.8 area yield kg-1) were suitable for dense planting and could be used to establish the raw material forest of SWH seed oil. The results of this study indicated the potential of development of selected SWH as novel oil crops and their wide cultivation.

Marine polysaccharides from Gelidium pacificum Okamura and Cereus sinensis reveal prebiotic functions.

Many marine polysaccharides as prebiotics can promote host health by modulating gut microbiota. This study investigated the beneficial effects of purified marine plant-derived Gelidium pacificum Okamura polysaccharide (GPOP-1) and marine animal-derived Cereus sinensis polysaccharide (CSP-1) on normal mice by modulating gut microbiota. The composition and diversity of gut microbiota were evaluated using 16S rRNA high-throughput sequencing. The results showed that GPOP-1 and CSP-1 altered the composition of the gut microbiota and promoted the growth of beneficial bacteria. At the genus level, GPOP-1 increased the relative abundance of Bacteroides, Phascolarctobacterium, and decreased the relative abundance of Ruminococcus, Helicobacter, Allobaculum, Dorea and AF12. While CSP-1 increased the relative abundance of Coprococcus, Adlercreutzia, Roseburia, Phascolarctobacterium, and decreased the relative abundance of Bacteroides, Ruminococcus and Oscillospira. The changes in the gut microbiota may affect the body weight, immune organ index and the production of short-chain fatty acids in normal mice. Compared to the normal control group, GPOP-1 decreased average weight gain while CSP-1 increased average weight gain. Furthermore, both GPOP-1 and CSP-1 significantly increased thymus and spleen indexes and total short chain fatty acids production in mice. In summary, GPOP-1 and CSP-1 exerted prebiotic effects on normal mice.

Isolation, structure identification and anti-inflammatory activity of a polysaccharide from Phragmites rhizoma.

Phragmites rhizoma (PR) is comprised of polysaccharides as its main active component. Currently, there are few studies on polysaccharides from PR, especially in purification and structure. In this study, an acidic polysaccharide (PRP-2) was obtained from PR by ultrasonic assisted extraction and secondary column chromatography purification (Diethylaminoethyl cellulose-52 (DEAE-52) and Sephadex G-100). Its structural characteristics were investigated by the gel permeation chromatography (GPC), gas chromatography and mass spectrometry (GC-MS), infrared spectroscopy (IR) and nuclear magnetic resonance (NMR) spectroscopy. The results showed that PRP-2 possessed the molecular weight of 20,332 Da and contained total sugars (71.73%), uronic acids (7.51%), proteins (0.57%) and sulfate radical (9.38%). The polysaccharide was composed of Galactose (34.70%), Fucose (36.15%) and a small amount of Rhamnose (0.88%) with a molar ratio of 39.50: 41.15: 1.00. It consisted of three sugar residues, →3)-ß-D-GalpA-(1→, →2, 3)-α-L-Fucp-(1→ and α-L-Fucp (4SO3-) -(1→. PRP-2 could protect RAW246.7 macrophages from the cytotoxic effect induced by lipopolysaccharide (LPS), inhibit the LPS-induced NO production in RAW246.7 macrophages, which displayed its anti-inflammatory activity. Therefore, PRP-2 has the potential to be used as a functional component.

Beneficial effects of sulfated polysaccharides from the red seaweed Gelidium pacificum Okamura on mice with antibiotic-associated diarrhea.

The purpose of this study was to investigate whether Gelidium pacificum Okamura polysaccharides (sulfated polysaccharide, GPOP-1) had beneficial effects on mice with antibiotic-associated diarrhea (AAD). Compared with the natural recovery group, GPOP-1 increased the richness and diversity of the gut microbiome, as well as altered the composition of the gut microbiota. At the genus level, GPOP-1 significantly increased the relative abundance of Bacteroides, Oscillospira, and Bifidobacterium and decreased the relative abundance of Parabacteroides, Sutterella, and AF12. The metabolic pathway differences according to the Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed that the metabolic function of the gut microbiota could be significantly improved by GPOP-1. Furthermore, GPOP-1 downregulated the concentrations of inflammatory cytokines, tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-2 (IL-2), alleviated the pathological features of the cecum, and increased the contents of acetates, propionates, butyrates, and total short-chain fatty acids (SCFAs). Results indicated that GPOP-1 had beneficial effects on mice with AAD by promoting the recovery of the gut microbiota and mucosal barrier function, reversing metabolic disorders, downregulating the levels of inflammatory cytokines and improving the content of SCFAs.

Fe3+-Coordinated Multifunctional Elastic Nanoplatform for Effective in Vivo Gene Transfection.

The common phenomenon that the nonviral vectors have much lower transfection efficiency in vivo than in vitro greatly restricts their further developments and applications. Possible reasons are lacking targeting ability, elimination by the reticuloendothelial system (RES), and insufficient nuclear transport. Here, a novel, flexible, and deformable polymer Fe@PEI-R12 (tLyp-1-NLS) is reported for shortening the gap between in vitro and in vivo gene transfection efficiency. The amorphous network structure Fe@PEI with deformation ability acquired by coordination cross-linking of Fe3+ and low-molecular-weight polyethylenimine (LMW-PEI) constructs the core and serves as the gene reservoir, and it can squeeze out through RES filter holes when trapped in the spleen. The bifunctional peptide R12 provided tumor targeting and enhanced nuclear delivery ability. Additionally, the Fe3+ from Fe@PEI-R12 could trigger endogenous hydrogen peroxide (H2O2) decomposition to produce O2, thereby reducing the adverse effects of tumor hypoxia. It is demonstrated that the Fe@PEI-R12/pDNA complexes could pass through membrane filters, subsequently achieving long circulation time, and Fe@PEI-R12 had a tendency to accumulate in tumor tissue and mediate pGL3-control expression. Therefore, the multifunctional nanoplatform has the potential for effective in vivo gene delivery.

Addition of free poloxamer 407 to a new gene vector P407-PEI-K12 solution forms a sustained-release in situ hypergel that enhances cell transfection and extends gene expression.

To address the concern around the efficiency/cytotoxicity ratio and the tumor-targeting effects of polyethylenimine (PEI), is a non-viral gene vector used for the delivery of the cancer therapy gene, poloxamer 407 (P407)-PEI-K12, was synthesized by cross-linking low-molecular weight PEI with P407 and further coupling a bifunctional peptide, K12, which is comprised of the tumor-targeting peptide tLyP-1 and the nuclear localization sequence. Furthermore, the addition of free P407 into the polymer/DNA complex solution produced a temperature-sensitive in situ gel-P407/P407-PEI-K12/DNA complex, which improved the effects of sustained-release gene delivery and transfection efficiency. The specificity, cytotoxicity and gene transfection efficiency of P407-PEI-K12 was investigated in Hela cells in vitro. The polymer efficiently prevented the degradation of plasmid DNA by DNase I and had a marked ability for serum tolerance. Agarose gel electrophoresis revealed that plasmid DNA was efficiently condensed and protected. The higher transfection efficiency of P407-PEI-K12h (the molar ratio of P407-PEI and K12 is 1:10) was achieved with a polymer and plasmid DNA ratio (w/w) of 20:1. The ability of free P407 to promote the transfection of the polymer/DNA complex was high (0.09%). The half-life of the P407/P407-PEI-K12-h/DNA gel complex was 228 min, and the transfection efficiency of the P407/P407-PEI-K12-h/DNA complex was markedly higher compared to that of the P407-PEI-K12-h/DNA complex at various release times.

Characterization and anti-inflammatory effects of sulfated polysaccharide from the red seaweed Gelidium pacificum Okamura.

In the present study, crude polysaccharides were extracted from Gelidium pacificum Okamura, and further purified to obtain the sulfated polysaccharide with molecular weight of 28,807 Da. Its monosaccharide composition mainly consisted of xylose (7.1%), galactose (59.7%) and galacturonic acid (19.76%). And the sulfate ester content of the sulfated polysaccharide was estimated as 8.8%. Structure analysis showed that the sulfated polysaccharide comprised of 1,4-linked-α-D-Galp3S, 1,2-linked-α-D-Xylp and 1,3-linked-ß-D-GalpA residues, respectively. Its anti-inflammatory effects were investigated in LPS-stimulated human monocytic (THP-1) cells. The sulfated polysaccharide at a concentration of 5 µg/mL fully protected the THP-1 cells against LPS-stimulated cytotoxicity. Furthermore, the addition of sulfated polysaccharide resulted in a significant reduction of NO production in LPS-treated cells, and this effect appeared to be dose-related. The sulfated polysaccharide (5 µg/mL) significantly suppressed the mRNA and protein expression of toll-like receptor-4 (TLR-4), myeloid differentiation factor (MyD88) and tumor necrosis factor receptor-associated factor-6 (TRAF-6) in LPS-stimulated THP-1 cells. These results showed the sulfated polysaccharide not only provided a good protection against LPS-induced cell toxicity, but also exerted an anti-inflammatory effect via the TLR4 signaling pathway.