Fig 2: Identification results of the main chemical components of XZR by UHPLC-LTQ-Orbitrap MS. (A) Total ion flow diagram of XZR in anion mode (details of Nos. 1–35 are listed in Supplementary Table S1). (B) Total ion flow diagram of XZR in the positive ion mode (details of Nos. 1–27 are listed in Supplementary Table S2). (C) Identification of the main components in XZR. XZR, Xiongshao Zhitong Recipe.

Fig 3: Administration of AMSCs improved intestinal structures, mucus secretion, tight junction, and peristalsis is reversed by spinal-origin neurotransmitter.(A–C) The ileum in PBS group (A) was more atrophic than those in the AMSC group (B) (magnification, ×100; scale bar: 200 µm), and the villus height in the AMSC group was significantly superior to those in the PBS group (C, P = 0.0118). (D–F) The ability to produce mucus was evaluated via PAS staining 3 days after SCI. The ileum in PBS group (D) had fewer goblet cells containing intestinal mucus than those in the AMSC group (E) (magnification, ×100; scale bar: 200 µm). The PAS-positive area in the AMSC group was significantly larger than that in the PBS group. (F; P = 0.0306). (G–H) The permeability between the intestinal epithelial cells was evaluated via IHC staining using the anti-zo-1 antibody. The zo-1 positive connections between the cells (arrows) in the PBS group (G) were less than those in the AMSC group (H) (magnification, ×100; scale bar: 200 µm). (I–K) Intestinal peristalsis was evaluated 14 d after SCI via the immunofluorescence staining using the anti-nNOS antibody. The nNOS-positive areas in the muscle layers in the PBS group (J) were significantly smaller than those in AMSC group (J) 14 d after SCI (K), (magnification, ×100; scale bar: 200 µm). Data are presented as mean ± standard error (SE). *, P<0.05; **, P<0.01.

Fig 4: Significant increase of Ntyr levels in neruons in enthorhinal cortex (EC) and hippocampal CA1 region of P301S Tg mice compared to WT controls.a, b Representative immunohistochemistry staining of 3-nitrotyrosine (Ntyr) (green) and DAPI (blue) for CA1 regions. c, d Co-localizing Ntyr with neurons (NeuN-Red). e, f Co-localizing Ntyr with astrocytes (GFAP-Red). g, h Co-localizing Ntyr with microglia (IBA-Red). i–j Representative staining of Ntyr (green) and DAPI (blue) for entorhinal cortex (EC) region. k Quantitative analysis of the fluorescence intensity of Ntyr in the CA1, CA2, CA3, dentate gyrus (DC), EC, prefrontal cortex (PFC), motor cortex (MC), and somatosensory cortex (SSC) regions. Values presented are the mean and ±SEM. One tailed t-test was conducted. **p < 0.005, WT mice (n = 6) and Tg mice (n = 6). l, m Representative WB (for nNOS) from cortex and hippocampus tissues prepared from Cor-WT, Cor-Tg, Hip-WT, and Hip-Tg. n The relative average WB intensity of nNOS normalized to GAPDH. WT mice (n = 3) and Tg mice (n = 3)

Fig 5: Changes in •NO synthases (NOS) protein expression was performed by Western blot analysis for neuronal NOS (nNOS) protein expression. Male—WT—Soy, n = 12; Male—WT—Soyfree, n = 8; Male—Tgß3—Soy, n = 6; Male—Tgß3—Soyfree, n = 11 (A), eNOS protein expression. Male—WT—Soy, n = 11; Male—WT—Soyfree, n = 8; Male—Tgß3—Soy, n = 6; Male—Tgß3—Soyfree, n = 11 (B) and iNOS protein expression. Male—WT—Soy, n = 3; Male—WT—Soyfree, n = 3; Male—Tgß3—Soy, n = 3; Male—Tgß3—Soyfree, n = 3 (C) on LV from male rats depending on the rodent diet (with or without soy at 45 weeks). n = Data are expressed as mean ± SEM *: p < 0.05, ***: p < 0.001.