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Ship ballast water contains high concentration of plankton, bacteria, and other microorganisms. If the huge amount of ballast water is discharged without being inactivated, it will definitely spell disaster to the marine environment. Microalgae is the most common species exiting in ballast water, so the detection of the concentration and viability of microalgae is a very important issue. The traditional methods of detecting microalgae in ballast water were costly and need the help of bulky equipment. Herein, a novel method based on microalgae cell intracellular chlorophyll fluorescence (CF) imaging combines with cell bright field (BF) microscopy was proposed. The geometric features of microalgae cells were obtained by BF image, and the cell viability was obtained by CF image. The two images were fused through the classic image registration algorithm to achieve simultaneous detection of the viability and concentration of microalgae cells. Furthermore, a low-cost, miniaturized CF/BF microscopy imaging prototype system based on the above principles was designed. In order to verify the effectiveness of the proposed method, four typical microalgae in ballast water (Platymonas, Pyramimonas sp., Chrysophyta, and Prorocentrum lima) were selected as the samples. The experimental results show that the self-developed prototype can quickly and accurately determine the concentration and the viability of microalgae cells in ship ballast water based on the dual images of BF and CF, and the detection accuracy is equivalent to that of commercial microscope. It was the first time to simultaneously detect the viability and concentration of microalgae cells in ship ballast water using the method that combining the fluorescence and bright field images, moreover, a miniaturized microscopic imaging prototype was developed. Those findings expected to contribute to the microalgae detection and ship ballast water management.

To identify bioactive components of Panax notoginseng (PN) roots in raw and steamed forms, chemometrics analyses including bivariate correlation analysis and partial least squares regression were used to establish the relationships between the chromatograms and anti-inflammatory effect of PN samples. The chemical fingerprints of PN were obtained by high performance liquid chromatography. The anti-inflammatory activity was evaluated by the TNF-α and IL-6 inhibiting test. According to the results, raw PN (RPN) displayed a stronger anti-inflammatory effect compared with steamed PN (SPN) samples. Chemometrics analyses showed that notoginsenoside R1, ginsenosides Rg1, Re and Rb1 were the major anti-inflammatory components of RPN, whereas ginsenosides 20(S)-Rg3 was the active one of SPN treating inflammation. The pharmacologic verification was consistent with the predicted results. In addition, the contents of anti-inflammatory components in RPN were higher than those in SPN, which might be the reason that RPN exhibited stronger effect on the inflammation induced by lipopolysaccharide in RAW264.7 cells. These findings could be beneficial for choosing differential markers for the quality control of RPN and SPN.

This work explored the feasibility of using biological polysaccharide to fabricate dissolvable microneedles (MNs) for the purpose of transdermal drug delivery and skin dendritic cell (DC) activation. Panax notoginseng polysaccharide (PNPS), a naturally derived immunoactive macromolecule, was used to fabricate dissolvable MNs. The prepared PNPS MNs showed a satisfactory mechanical strength and a skin penetration depth. By Franz diffusion cell assay, the PNPS MNs demonstrated a high transdermal delivery amount of model drugs. Furthermore, with the assistance of MNs, PNPS easily penetrated across the stratum corneum and target ear skin DCs, activating the maturation and migration of immunocytes by increasing the expressions of CD40, CD80, CD86, and MHC II of skin DCs. Consequently, the matured DCs migrated to the auricular draining lymph nodes and increased the proportions of CD4+ T and CD8+ T cells. Thus, PNPS might be a promising biomaterial for transdermal drug delivery, with adjuvant potential.

In the current study, we developed a synergistic chemo-immunotherapy using doxorubicin (Dox) and a natural polysaccharide as immunomodulator. First, we isolated a polysaccharide (MPW) from the root of Lepidium meyenii Walp. (maca) and characterized its chemical properties. MPW contains → 4) -α-D-Glcp- (1 → glycosidic bonds, while the terminal α-D-Glcp- (1 → group is connected to the main chain through an O-6 bond. This polysaccharide was then modified by cationization (C-MPW) to enhance immunoregulatory activity. MPW and C-MPW were combined with Dox and their chemo-immunotherapy effects on 4T1 tumor-bearing mice were assessed. Results indicated that the combination of MPW/C-MPW exerted a stronger anti-tumor effect than Dox alone, while reducing systemic toxicity and inhibiting tumor metastasis. In addition, MPW and C-MPW exerted tumor immunotherapy effects through the NF-κB, STAT1, and STAT3 signaling pathways, redirecting TAMs to the M1 phenotype that facilitates immunological responses against tumors. As a result, the immunosuppressive tumor microenvironment was remodeled into an immune-activated state due to enhanced secretion of IL-12, TNF-α, and INF-γ. Moreover, C-MPW exerted a stronger immunomodulatory effect than MPW. In conclusion, MPW and its cationic derivative are promising tools for cancer immunotherapy.

This study isolated and characterized a novel polysaccharide (PNPS-0.3) from the residue of Panax notoginseng by gradient elution. PNPS-0.3 mainly consisted of a backbone of →4)- α-D-GalAp-(1 → 4-β-L-Rhap-1 → 4)-β-D-Galp-(1 → residues, with an α-L-Araf-1 → 5)-α-L-Araf-(1 → branch connecting to the backbone at O-3 of →4-β-L-Rhap-1 → and a molecular weight of 76,655 Da. Furthermore, the adjuvant potential of PNPS-0.3 with bone marrow dendritic cells (BMDCs) was investigated. The results suggested that PNPS-0.3 could induce maturation of BMDCs by reshaping the morphology, upregulating the CD40, CD80, CD86 and MHC II membrane phenotypic markers, and by promoting the secretion of TNF-α and IL-12 proinflammatory cytokines. Moreover, PNPS-0.3 can trigger the DC-induced T-cell immune response, as indicated by the higher expressions of CD4, CD8, CD69, and MHC II in T cells with increased secretion of INF-β. Furthermore, PNPS-0.3 can bind to the pattern recognition receptors (PRR) of Toll-like receptor 4 (TLR 4), Toll-like receptor 2 (TLR 2), and mannose receptor (MR) on BMDCs. PNPS-0.3 also upregulated the expressions of Myd88, IKKβ, PP65, T-P65, and NF-κB, suggesting that the TLR4/TLR2-NF-κB signaling pathway was involved in the immunomodulatory mechanism. In conclusion, the immunoadjuvant potential of novel PNPS-0.3 was characterized, which is beneficial for the future utilization and development of P. notoginseng.

In this investigation, innovative pH-sensitive and amphiphilic nanoparticles (NPs) were synthesized by grafting histidine (His, pH sensitive molecule) and stearic acid (SA, hydrophobic segment) onto the polysaccharides of Bletilla striata (BSP). The His-SA-BSP was able to self-assemble into NPs with pH sensitivity. The acidic conditions could trigger the imidazole ionization and reverse the surface charge, while the electrostatic repulsion wrecked the structure and drove the NPs to a swollen state, as revealed by dynamic light scattering (DLS), transmission electron microscopy (TEM), and critical micelle concentration (CMC) analyses. By increasing the degree of substitution (DS) of His, the NPs showed improved pH sensitivity. The NPs could accelerate Doxorubicin (Dox) release to a remarkably greater extent (3-fold) at pH 5 than at pH 7.4. The CCK-8 assay demonstrated a good biocompatibility of the NPs towards different cell lines and a specific inhibition effect of Dox-loaded NPs against tumor cells. Furthermore, the NPs showed the improved cellular uptake of Dox towards MCF-7 by fluorescence microscopy and flow cytometry. Therefore, the new His-SA-BSP showed potential applications in drug nanocarrier systems.

This study developed novel ionic liquids (ILs) based on amino acids. We first screened 15 methyl amino acid ester hydrochlorides ([AAC1]Cl) for their skin permeation enhancements using 5-Fluorouracil (5-Fu) and Hydrocortisone (HC) as model drugs. Glycine methyl ester hydrochloride ([GlyC1]Cl), L-proline methyl ester hydrochloride ([L-ProC1]Cl), and L-leucine methyl ester hydrochloride ([L-LeuC1]Cl) were selected, and their ester sites were modified with different carbon chains (C8 and C12). The resulting ILs showed improved permeation to the two drugs. TEWL and CLSM assays revealed the moderation effects of the modified ILs on skin barrier function, whereas L-proline dodecyl ester hydrochloride ([ProC12]Cl) and L-leucine dodecyl ester hydrochloride ([L-LeuC12]Cl) exhibited the strongest activities. Permeation mechanisms were further investigated by ATR-FTIR, solid-NMR, SEM, and TEM analyses. The results suggested that [L-ProC12]Cl and [L-LeuC12]Cl combined the advantages of amino acid esters and IL solvent and could interact with the intercellular lipid domain by the multi-functions of lipid fluidization and lipid extraction, which were observed in a dosage- and time-dependent manner. Additionally, pathological examination suggested that the amino acid ester-based ILs (AAE-ILs) had good biocompatibility. In conclusion, this study has generated novel chemical penetration enhancers (CPEs) based on AAE-ILs and may be potentially utilized in drug transdermal delivery systems (TDDSs).

This study presented the first purification and characterization of a hepatoprotective polysaccharide (PNPS-0.5 M) from the residue of Panax notoginseng (Burk.) F.H. Chen. This polysaccharide included a backbone of (4 → 1)-linked GalA and branches of (1→)-linked Araf, (1→)-linked Rhap, and (5 → 1)-linked Araf and had an extremely high molecular weight (2600 kDa). We investigated the hepatoprotective effects of PNPS-0.5 M on mice with alcoholic liver damage (ALD). After administration of PNPS-0.5 M, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), and hepatic malondialdehyde (MDA) were reduced to normal. In contrast, hepatic levels of alcohol dehydrogenase (ADH) and the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were elevated to normal. Further investigations indicated that PNPS-0.5 M activated Nrf2 signaling as a protective mechanism against Cyp2e1 toxicity in ALD mice. Meanwhile, it strengthened the ADH pathway and suppressed the CAT pathway of alcohol metabolism to prevent peroxide accumulation, thereby ameliorating ALD. In the present study, we describe a novel acidic polysaccharide from P. notoginseng with hepatoprotective activity that facilitates the development and utilization of P. notoginseng resources.

In this work, imidazolium ionic liquids (imidazolium ILs) were employed as the novel chemical permeation enhancers (CPEs) and their performances and mechanisms of action were deeply investigated. Testosterone was used as a model drug to investigate the transdermal delivery enhancement of twenty imdidazolium ILs. The results suggested that the promotion activity connected to the structure and composition of the ILs. The quantitative structure-activity relationship (QSAR) model revealed a good linearity between the electronic properties of ILs and their enhancements. Furthermore, the transepidermal water loss (TEWL) and scanning laser confocal microscope (CLSM) examinations showed the strong improvement of ILs on skin barrier permeability, which were well correlated with the drug penetration profiles. The total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) and atomic force microscope (AFM) evaluations of skins indicated that the ILs can disrupt the regular and compact arrangements of the corneocytes, change the surface properties of stratum corneum, and make the skin structure more permeable. Our work demonstrated the significant skin permeation promotion profiles of the imidazolium ILs, which are of great potential in transdermal drug delivery systems.

The Panax notoginseng (P. notoginseng) stem leaf is rich in flavonoids. However, because of a lack of research on the flavonoid extraction process and functional development of P. notoginseng stem leaf, these parts are discarded as agricultural wastes. Therefore, in this study, we intend to optimize the extraction process and develop the skin-whitening functions of P. notoginseng stem leaf extracts. The extraction process of the stem and leaf of P. notoginseng flavonoid (SLPF) is optimized based on the Box&ndash, Behnken design (BBD) and the response surface methodology (RSM). The optimum extraction conditions of the SLPF are as follows: the extraction time, the ethanol concentration, the sodium dodecyl sulfate (SDS) content and the liquid material ratio (v/w, which are 52 min, 48.7%, 1.9%, and 20:1, respectively. Under the optimal extraction conditions, the average total SLPF content is 2.10%. The antioxidant activity and anti-deposition of melanin of mouse B16 cells of P. notoginseng stem leaf extracts are studied. The results indicate that the EC50 values of reducing activity, 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activities, the superoxide anion removal ability, and the 2,2-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) free radical removal ability are 7.212, 2.893, 2.949, and 0.855 mg/mL, respectively. The extracts IC50 values of the tyrosinase and melanin synthesis are 0.045 and 0.046 mg/mL, respectively. Therefore, the optimal processing technology for the SLPF obtained in this study not only increases its utilization rate, but also decreases material costs. The extracts from the P. notoginseng stem leaf may be developed as food or beauty products.

A novel microemulsion was developed and characterized for topical delivery of Dencichine (Den). Two imidazaolium ionic liquid, 1-hydroxyethyl-3-methylimidazolium chloride ([HOEIM]Cl) and 1-butyl-3-methylimidazolium dodecanesulfate ([BMIM]C12SO3) were incorporated into the aqueous and surfactant phases respectively for the remarkable enhancement on skin permeation. The nano-carrier was developed and optimized based on a pseudo-ternary phase diagram. The optimized formulation was composed of 50% water/[HOEIM]Cl mix (1:1) as water phase, 20% Tween 80/[BMIM]C12SO3 mix (1:1) as surfactant, 10% propylene glycol as co-surfactant and 20% IPM as oil phase. The o/w microemulsion was then characterized for droplets sizes (47.7±1.5nm), zeta potential (-14.83±3.64mV), viscosity (31±4 mPa) and pH (6.71±0.04). In-vitro skin permeation assay suggested the strong enhancement of ILs formulation on the topical delivery of Den, which was approximately 10-fold that of the drug aqueous solution. It was found that the nano-carrier can reduce the skin barrier properties by disrupting the regular and compact arrangements of corneocytes, and moderating the surface properties of the stratum corneum, as evidenced by Transdermal Water Loss Evaluation (TEWL), Differential Scanning Calorimetery (DSC) and attenuated total Reflectance Fourier Transform Infrared spectroscopy (ATR-FTIR). Furthermore, the in-vivo pharmacodynamic evaluation indicated the significant hemostatic activity of Den by the topical application of the vehicle. Additionally, the formulation showed minor cell toxicity and skin irritation. Therefore, our work suggested that the ionic liquid microemulsion can be a promising nano-scale vehicle for the topical application of Den to produce desirable pharmacological effects.

Natural polysaccharides constitute promising material for nano-drug delivery systems (NDDS) due to their excellent biodegradability and biocompatibility. Previously, novel copolymer micelles (His-SA-BSP) were successfully engineered based on the functionalization of Bletilla striata polysaccharides (BSP) by histidine (His) and stearic acid (SA). In this work, the effectiveness of the designed micelles in selectively delivering Doxorubicin (Dox) to tumor cells is evaluated. The obtained results show satisfactory encapsulation efficiency, loading capacity, and drug compatibility of the copolymers. Surface charge conversion and micelle deterioration were observed at pH = 6.5 and pH = 5.0, respectively. In vitro Dox release assays indicate a rapid release of Dox from micelles under acidic conditions of tumor microenvironments. Furthermore, in vitro cytotoxicity and cellular uptake assays performed on Michigan Cancer Foundation-7 (MCF-7) cells suggest that the proposed micelles are safe to use in biological systems, and that they can be easily and selectively accumulated at the tumor site due to pH sensitivity. Endocytosis inhibition and intracellular delivery assays reveal that BSP nanocarriers are mainly delivered to lysosomes by micropinocytosis. The acidic condition of lysosomes (pH = 5.0) then triggers the disintegration micelles, resulting in a fast release of Dox to the nuclei. Finally, in vivo biodistribution and antitumor assessments conducted on MCF-7-BALB/c tumor xenograft mice confirm the enhanced tumor accumulation and antitumor efficacies of the investigated Dox-loaded micelles. In conclusion, the pH-sensitive drug delivery systems designed previously are shown to yield improved accumulation of the drug at the tumor site, enhanced intracellular uptake, and efficient in vivo antitumor efficacy.

In this study, the extraction process of Maca polysaccharide (LMP) was deeply studied and its chemo‐protective effects on Cyclophosphamide (Cy)‐induced mice were evaluated. By single factor investigation and response surface methodology optimization, it was found that the liquid‐to‐solid ratio, extraction time, and extraction temperature were the main factors which affected the extraction yield of LMP. The optimum extraction process was obtained by Box–Behnken design combined with response surface methodology. It was found that, at the condition of liquid‐to‐solid ratio of 30:1; extraction time of 2.3 hr; and extraction temperature of 90°C, the yield of the LMP could reach 46.56%. Furthermore, by oral administration of LMP, the cyclophosphamide‐induced mice showed the proliferation of lymphocyte and the phagocytic capacity of macrophages’ enhancements. The number of white blood cell, red blood cell, and platelets in the peripheral blood were up‐regulated; the spleen total superoxide dismutase, Catalase, and Glutathione‐peroxidase activities were enhanced, while the Serum malondialdehyde and Triacylglycerols content was reduced. Therefore, our work provided an effective way for extraction of LMP and demonstrated the chemo‐protective effects of LMP on the animal level. PRACTICAL APPLICATION: Natural polysaccharides from functional food sources are widely accepted for their bioactive activities. As extraction conditions significantly affected the quality and yield of polysaccharides, the effective extraction process for polysaccharides should be systematically investigated. In the current work, the optimum conditions for polysaccharides extraction from Maca (Lepidium meyenii Walp.) (LMP) were obtained using liquid‐to‐solid ratio of 30:1 for 2.3 hr at 90°C. The Mass characterization and Monosaccharide Composition of LMP were also determined. Furthermore, the extracted LMP exhibited well Chemo‐protective activities on Cyclophosphamide‐induced mice. Therefore, our work showed the potential applications of Maca polysaccharides as the easily accessible source of natural immunopotentiating agents in function food and pharmaceutical industry.