Additional file 1 of Analgesic effect of nitrous oxide during manual therapy after anterior cruciate ligament reconstruction: a study protocol for a randomized controlled trial

Additional file 1: [S1. SPIRITR2]. Standard Protocol Items: Recommendations for Interventional Trials (SPIRIT) Checklist

Light Extraction of Trapped Optical Modes in Polymer Light-Emitting Diodes with Nanoimprinted Double-Pattern Gratings

Despite the rapid development of polymer light-emitting diodes (PLEDs), the overall device efficiency is still limited because ∼80% of the generated light is trapped in a conventional device architecture by the high refractive index of organic materials and the optical confinement and internal reflection. The implementation of the energy dissipation compensation techniques is urgently required for further enhancement in the efficiency of PLEDs. Here, we demonstrate that incorporating the double-pattern Bragg gratings in the organic layers with soft nanoimprinting lithography can dramatically enhance the light extraction of trapped optical modes in PLEDs. The resulting efficiency is 1.35 times that of a conventional device with a flat architecture used as a comparison. The experimental and theoretical analyses indicate that the enhanced out-coupling efficiency is attributed to the combination of the ordinary Bragg scattering, the guided-mode resonance (GMR), surface plasmon polariton (SPP) modes, and the hybrid anticross coupling between GMR and SPP, leading to the extraordinary efficient photo flux that can transfer in direction of the leaky modes. We anticipate that our method provides a new pathway for precisely manipulating nanoscale optical fields and could enable the integration of different optical modes in PLEDs for the viable applications

ADAM19 was a direct target of miR-30c.

<p>(A) The predicted targeting site with miR-30c of ADAM19 3’-UTR; MiR-30c targeting sequences of ADAM19 3’-UTR are evolutionarily conserved through eight species (Hsa: human; Ptr: chimpanzee; Mml: rhesus; Oga: bushbabay; Tbe: treeshrew; Mmu: mouse; Rno: rat; and Ocu: rabbit.). The targeting sites are highlighted in red. (B-C) qRT-PCR and western blot analysis was applied to detect mRNA and protein expression of ADAM19 in HCT116 cells after transfection of miR-30c mimic or NC mimic. (D) ADAM19 mRNA and miR-30c expression level was determined in five colon cancer cell lines by qRT-PCR. (E) ADAM19 mRNA and miR-30c levels were inversely correlated in colon cancer tissues as determined by qRT-PCR. β-actin and U6 were used as the endogenous controls, respectively. (F) HEK293T cells were co-transfected with wild-type or mutant reporters and the miR-30c mimic or negative control (NC mimic). After 48h, Luciferase/Renilla activity was measured. (G) ADAM19 was upregulated in colon cancer tissues compared with normal tissues, as indicated by Western blotting.</p

MiR-30c inhibited cell migration and invasion ability of colon cancer cells.

<p>(A-B) Effects of miR-30c on migration were analyzed by Transwell migration assay. A, representative photos; B, quantitative analysis. (C-D) Effects of miR-30c on invasion were analyzed by Transwell invasion assay. C, representative photos; D, quantitative analysis. (E-F) F, Wound healing assay was adopted to evaluate the effect of miR-30c on migration. The artificial gap was through the central axis when cells reached a density of 80%. Photos of cells were taken at 0 and 24 h. E, representative photos; F, quantitative analysis. *<i>p</i><0.05; ** <i>p</i><0.01.</p

MiR-30c inhibited cell proliferation in vitro.

<p>(A) Relative expression levels of miR-30c in five colon cancer cell lines were detected with the quantitative real-time PCR (qRT-PCR). Columns, average of three independent experiments; bars, S.E. (B) HCT116 or Lovo cells were transiently transfected with miR-30c mimic or NC mimic and inhibitor or NC inhibitor respectively. The expression of miR-30c was determined by qRT-PCR after 24h. (C-D) Effects of miR-30c on the proliferation were examined by MTT assay. Points are the average of three independent experiments; bars represent S.E.; *<i>p</i><0.05; ** <i>p</i><0.01. (E) Effects of miR-30c on the proliferation of HCT116 cells were examined by colony formation assay. (F) The number of clones was quantitatively analyzed. *<i>p</i><0.05; **<i>p</i><0.01.</p

Primers for plasmid construction, qRT-PCR, and oligonucleotide.

<p>Primers for plasmid construction, qRT-PCR, and oligonucleotide.</p

MiR-30c inhibited tumor growth in vivo.

<p>(A) The stable overexpression of miR-30c in HCT116 cell clones was determined with qRT-PCR. (B) 5×10⁶ HCT116 cells which were stably transfected with miR-30C or empty vector (mock) were subcutaneously injected into nude mice (n = 3). Mice were sacrificed 28 days after injection. (C) Tumors were harvested and representative tumors were shown. (D) Tumors were weighted and the miR-30C overexpression group had a lower weight compared with the mock group. (E) MiR-30c overexpression resulted in inhibition of growth rate. (F) The expression of miR-30c was detected with qRT-PCR in tumors. (G-H) Hematoxylin and eosin staining of sections of lung and statistical results of metastasis nodes. ** <i>p</i><0.01.</p

Characteristics and miR-30c expression in colon cancer patients.

<p>* The correlation between miR-30c and CEA and CA199 is determined by Spearman’s correlation.</p><p>Characteristics and miR-30c expression in colon cancer patients.</p

MiR-30c was down-regulated in colon cancer tissues.

<p>MiR-30c expression was examined by qRT-PCR in 60 pairs of human colon cancer tissues. 54 out of 60 tumor samples showed decreased expression of miR-30c compared with the adjacent normal mucosa tissues. MiR-30c expression was normalized to that of U6 in each sample.</p