Additional file 1 of Effects of Enteromorpha prolifera polysaccharides on growth performance, intestinal barrier function and cecal microbiota in yellow-feathered broilers under heat stress

Additional file 1: Table S1. The chemical and monosaccharide composition of seaweed-derived polysaccharides (SDP) from Enteromorpha prolifera. Table S2. The formulation and nutrient level of the basal diets. Table S3. Primer information of real-time quantitative PCR

Vitamin E Succinate-Grafted-Chitosan Oligosaccharide/RGD-Conjugated TPGS Mixed Micelles Loaded with Paclitaxel for U87MG Tumor Therapy

The poor therapeutic efficacy of hydrophobic chemotherapeutic drugs is an intrinsic limitation to successful chemotherapy. In the present study, a multitask delivery system based on arginine-glycine-aspartic acid peptide (RGD) decorated vitamin E succinate (VES)-grafted-chitosan oligosaccharide (CSO)/RGD-conjugated d-alpha-tocopheryl polyethylene glycol 1000 succinate (TPGS-RGD) mixed micelles (VeC/T-RGD MM) was first prepared for targeted delivery of a hydrophobic anticancer drug, paclitaxel (PTX), to improve the efficacy of U87MG tumor therapy. VES grafted CSO (VES-<i>g</i>-CSO) and TPGS-RGD were synthesized as nanocarriers, and PTX loaded VeC/T-RGD MM (PTX@VeC/T-RGD MM) was prepared via the organic solvent emulsification–evaporation method. The PTX@VeC/T-RGD MM was 150.2 nm in diameter with uniform size distribution, 5.92% drug loading coefficient, and no obvious particle size changes within 7 days. The PTX@VeC/T-RGD MM showed sustained-release properties <i>in vitro</i> and high cytotoxicity, and could be efficiently taken up by human glioma U87MG cells. The tumor inhibitory rate of PTX@VeC/T-RGD MM treatment in U87MG tumor spheroids and U87MG tumor bearing mice was 49.3% and 88.4%, respectively, which indicated a superior therapeutic effect. PTX@VeC/T-RGD MM did not damage normal tissues in safety evaluations. These findings suggested that PTX@VeC/T-RGD MM could be developed for the delivery of hydrophobic drugs to U87MG tumors

CSO-g-CM-β-CD synthesis and characterization.

<p>Synthetic route of CSO-g-CM-β-CD (A); FT-IR spectra of CSO-g-CM-β-CD and CSO (B) and ¹H-NMR spectra of CSO and CSO-g-CM-β-CD in D₂O (C).</p

The influence of pH on CSO-g-CM-β-CD@AD-PTX micelle formation.

<p>The influence of pH on CSO-g-CM-β-CD@AD-PTX micelle formation.</p

<i>In vitro</i> drug release of CSO-g-CM-β-CD@AD-PTX micelle in the PBS with pH = 7.4 (n = 3).

<p><i>In vitro</i> drug release of CSO-g-CM-β-CD@AD-PTX micelle in the PBS with pH = 7.4 (n = 3).</p

Fluorescence analysis.

<p>Fluorescence scanning analysis with pyrene (A) and critical micelle concentration of CSO-g-CM-β-CD@AD-PTX solution (B).</p

<i>In vitro</i> cell viability assay of different formulations on U87 MG cells for 72 h (n = 3).

<p><i>In vitro</i> cell viability assay of different formulations on U87 MG cells for 72 h (n = 3).</p

AD-PTX synthesis and characterization.

<p>Synthetic route of AD-PTX (A); LC-MS spectra of AD-PTX (B) and ¹H-NMR spectra of AD, PTX and AD-PTX in DMSO-d₆ (C).</p

Characterization of CSO-g-CM-β-CD@AD-PTX micellar system.

<p>TEM images (A) and size distribution of CSO-g-CM-β-CD@AD-PTX micelles (B).</p

Effect of MG132 on VEGF and p53 expression induced by active Rac1/Cdc42.

<p>(A) GST–MCF-7, V12Rac1–MCF-7, and L61Cdc42–MCF-7 cells were incubated, and MG132 was added to select samples for 12 and 24 h, respectively. After incubation, the media were analyzed for VEGF levels using ELISA assay; cell numbers were counted. Data are expressed as the mean ± standard deviation (SD) for three independent experiments. Statistical significance was assessed with one-way ANOVA and Student's t-test. *<i>P</i><0.05, **<i>P</i><0.01, for the GST–MCF-7 group vs V12Rac1– or L61Cdc42–MCF-7 group. #<i>P</i><0.05, ##<i>P</i><0.01, for the MG132-added group vs the no-MG132-added group. (B, C) MCF-7, GST–MCF-7, V12Rac1–MCF-7 and L61Cdc42–MCF-7 cells were incubated, and MG132 was added to select samples for 24 h. After incubation, the cells were analyzed by western blot assay. p53, p21 and β-actin expression was examined. β-actin protein levels were used as a loading control. (D) The immunoprecipitation assay was performed as described in Material and Methods. Total protein was extracted from MCF-7, GST–MCF-7, V12Rac1–MCF-7 and L61Cdc42–MCF-7 cells and subjected to an immunoprecipitation assay. p53 protein expression was examined. β-actin protein levels were used as a loading control.</p