Additional file 3: Figure S2. of C-C motif chemokine ligand 20 regulates neuroinflammation following spinal cord injury via Th17 cell recruitment

The temporal profile (from 0 h to 28 days post-injury) of IL-17A mRNA expression in the spinal cord. IL-17A, as determined by qRT-PCR, shows that SCI leads to increased IL-17A mRNA level in the spinal cord, especially at 14 days post-SCI. + P < 0.05, compared with the sham group. (DOCX 168 kb

Additional file 1: Figure S1. of C-C motif chemokine ligand 20 regulates neuroinflammation following spinal cord injury via Th17 cell recruitment

Animal model of contusion SCI. (A) The localization of T10 by X-ray. (B) Skin preparation. (C) Skin incision. (D) The exposure of T10. (E) The exposure of the spinal cord. (F) Contusion injury. (G) Observation after SCI. (H) Suture the incision. (DOCX 2344 kb

Results (<i>F</i> and <i>P</i> values) of a three-way ANOVA on the effects of year (Y), nitrogen addition (N), water addition (W), and their interactions on aboveground biomass (AGB, g m^-2), net ecosystem CO₂ exchange (NEE, μmol m^-2 s^-1), ecosystem respiration (ER, μmol m^-2 s^-1), and gross ecosystem productivity (GEP, μmol m^-2 s^-1).

<p>The bold numerals highlight significance at the <i>P</i> < 0.05 level.</p><p>Results (<i>F</i> and <i>P</i> values) of a three-way ANOVA on the effects of year (Y), nitrogen addition (N), water addition (W), and their interactions on aboveground biomass (AGB, g m^-2), net ecosystem CO₂ exchange (NEE, μmol m^-2 s^-1), ecosystem respiration (ER, μmol m^-2 s^-1), and gross ecosystem productivity (GEP, μmol m^-2 s^-1).</p

Responses of net ecosystem CO₂ exchange (NEE, μmol m^-2 s^-1) to monthly precipitation regimes (A, frequency; B, amount; C, intensity; D, evenness) across the 3 years for both nitrogen fertilized (open circles, A: y = Exp(2.240 – 2.662x^-1); B: y = Exp(2.017 – 6.269x^-1); C: y = Exp(2.053 – 1.190x^-1); D: y = Exp(2.479 – 0.316x^-1)) and unfertilized plots (solid circles, A: y = Exp(1.905 – 3.610x^-1); B: y = Exp(1.613 – 9.101x^-1); C: y = Exp(1.700 – 1.983x^-1); D: y = Exp(2.141 – 0.381x^-1)).

<p>Nitrogen fertilized plots include both N and WN plots, and unfertilized plots include both CK and W plots. Water addition was treated as a precipitation event and included in the calculation of monthly precipitation regimes. * and ** represent significant relationships at the <i>P</i> < 0.05 and <i>P</i> < 0.01 levels, respectively.</p

Dependence of growing season mean (A) net ecosystem CO₂ exchange (NEE, μmol m^-2 s^-1), (B) ecosystem respiration (ER, μmol m^-2 s^-1), and (C) gross ecosystem productivity (GEP, μmol m^-2 s^-1) on aboveground biomass (AGB) across treatments in 2012 (solid circles), 2013 (open triangle), and 2014 (open square).

<p>** represents a significant relationship at the <i>P</i> < 0.01 level.</p

Seasonal patterns of (A) daily precipitation (bars, mm), daily mean air temperature (lines, °C), and (B) monthly precipitation (mm) in 2012, 2013, and 2014.

<p>Arrows on top of panel A indicate dates when water addition treatments were applied.</p

Positive dependence of growing season mean (A) net ecosystem CO₂ exchange (NEE, μmol m^-2 s^-1), (B) ecosystem respiration (ER, μmol m^-2 s^-1) and (C) gross ecosystem productivity (GEP, μmol m^-2 s^-1) on the amount of early growing season (Apr, May, Jun) precipitation (water added + natural precipitation) across the 3 years in both fertilized (open circles) and unfertilized plots (solid circles).

<p>Nitrogen fertilized plots include both the N and WN plots, and unfertilized plots include both the CK and W plots. Water addition was treated as a precipitation event and included in the calculation of the precipitation amount. * and ** represent significant relationships at the <i>P</i> < 0.05 and <i>P</i> < 0.01 levels, respectively.</p

Seasonal dynamics and means of (A, B, and C) net ecosystem CO₂ exchange (NEE, μmol m^-2 s^-1), (D, E, and F) ecosystem respiration (ER, μmol m^-2 s^-1), and (G, H, and I) gross ecosystem productivity (GEP, μmol m^-2 s^-1) in 2012, 2013, and 2014.

<p>Different lowercase letters indicate significant differences (<i>P</i> < 0.05) in seasonal averages among treatments (Duncan’s test). CK: control; W: water addition; N: nitrogen addition; WN: water and nitrogen added in combination. Data are reported as mean ± 1 SD (<i>n</i> = 6).</p

Seasonal means of soil pH (unitless value), vegetation density of the community (VD, plant m^-2), <i>Leymus chinensis</i> importance value (IV), aboveground biomass (AGB, g m^-2), and belowground (to 10 cm depth) biomass (BGB, g m^-2) under different treatments in July across the 3 years.

<p>Values are presented as mean ± 1 SD (<i>n</i> = 6). Different letters in a column indicate significant difference among treatments at the <i>P</i> < 0.05 level (Duncan test). CK: control; W: water addition; N: nitrogen addition; WN: water and nitrogen added.</p><p>Seasonal means of soil pH (unitless value), vegetation density of the community (VD, plant m^-2), <i>Leymus chinensis</i> importance value (IV), aboveground biomass (AGB, g m^-2), and belowground (to 10 cm depth) biomass (BGB, g m^-2) under different treatments in July across the 3 years.</p

Dual Stimuli-Responsive Nanoparticle-Incorporated Hydrogels as an Oral Insulin Carrier for Intestine-Targeted Delivery and Enhanced Paracellular Permeation

For enhanced oral insulin delivery, a strategy of acid-resistant and enteric hydrogels encapsulating insulin-loaded nanoparticles was developed. The nanoparticles were prepared by the formation of an anionic insulin/heparin sodium (Ins/HS) aggregate, followed by coating of chitosan (CS) on the surface. The nanoparticles, tagged as CS/Ins/HS NPs, exhibited excellent mucosa affinity, effective protease inhibition, and marked paracellular permeation enhancement. Moreover, to improve the acid-stability of CS/Ins/HS NPs and impart the capacity of intestine-targeted delivery, a pH- and amylase-responsive hydrogel was synthesized via free radical copolymerization, using methacrylic acid as the monomer and acrylate-<i>grafted</i>-carboxymethyl starch as the cross-linker. The resulting hydrogel exhibited sharp pH-sensitivity in the gastrointestinal tract and rapid enteric behavior under intestinal amylase. The additional protection for insulin in artificial gastric fluid was confirmed by packaging CS/Ins/HS NPs into the hydrogel. The obtained nanoparticle-incorporated hydrogel was named as NPs@Gel-2. The release of insulin from NPs@Gel-2 was evidently accelerated in artificial intestinal fluid containing α-amylase. Furthermore, the hypoglycemic effects were evaluated with type-1 diabetic rats. Compared to subcutaneous injection of insulin solution, the relative pharmacological availability (rPA) for oral intake of NPs@Gel-2 (30 IU/kg) was determined to be 8.6%, along with rPA of 4.6% for oral administration of unpackaged CS/Ins/HS NPs (30 IU/kg). Finally, the two-week therapeutic outcomes in diabetic rats were displayed after twice-daily treatments by oral intake of NPs@Gel-2, showing the relief of diabetic symptoms and suppression of weight loss in the rats. Therefore, this dual stimuli-responsive nanoparticle-incorporated hydrogel system could be a promising platform for oral insulin delivery