Summary of pre- and postoperative SSA(°) in relation to the levels involved.

<p>No., number; SSA, sagittal segmental alignment; Preop, preoperative; Postop, postoperative.</p

Summary of pre- and postoperative SACS(°) in relation to the levels involved.

<p>No., number; SACS, sagittal alignment of the whole cervical spine; Preop, preoperative; Postop, postoperative.</p

A 52-year-old female who underwent 1-level corpectomy and TMC fusion.

<p><b>2A.</b> Postoperative lateral X-ray on the second day after operation showed detectable gap between anterior plate and upper vertebral body. <b>2B.</b> Postoperative lateral X-ray at the 5-year follow-up showed noticeable screw loosening at the upper vertebral body and spontaneous fusion of heterotopic ossification at the adjacent level.</p

Overexpression of miR-27a in nucleus pulposus cells, and its function in the regulation of target proteins

<p>(A) Comparison of cell proliferation in various NP cell groups (*P<0.05). Data are representative of three experiments; error bars represent SEM. (B) Increased expression of cleaved caspase 3 and decreased expression of PIK3CD in the group overexpressing miR-27a compared to the control group (magnification ×200), respectively. Black arrows indicate positive-stained cells. (C) Contour diagram of FITC-Annexin V/PI FCM of human NP cells. The graphs represent typical results of cellular apoptosis; values represent the means of three experiments. Error bars represent SEM. (D) Results of cellular apoptosis was expressed as a fold change. Results are shown as mean ± SEM. Data are representative of three independent experiments (*P<0.05, ***P<0.001). (E) Increased numbers of nucleus pulposus cells were observed to undergo apoptosis in the group overexpressing miR-27a compared to the control group (magnification ×200). (F) (G) After 36 h, cellular protein lysates were prepared and PIK3CD expression was assessed by Western blot. GAPDH was used as an internal loading standard. Results are shown as mean ± SEM. Data are representative of three independent experiments (*P<0.05, **P<0.01).</p

MiR-27a can inhibit PIK3CD by targeting the 3’-UTRs of PIK3CD.

<p>(A) Complementarity between miR-27a and the putative PIK3CD 3’-UTR target site. PIK3CD 3’-mut indicates the PIK3CD 3’-UTRs with three mutation sites (underlined) in miR-27a binding sites. (B) The relative luciferase activities of three independent experiments are shown. Error bars represent SEM; (**P<0.01).</p

The expression levels of miR-27a were analyzed in human degenerative NP compared with control NP by real-time RT-PCR.

<p>The relative expression of miR-27a was normalized to the endogenous control U6. Each sample was analyzed in triplicate (**<i>P</i> <0.01, ***<i>P</i><0.001). (A) miR-27a is up-regulated in damaged NP <i>in </i><i>vitro</i>. (B) miR-27a is up-regulated in damaged NP cells <i>in </i><i>vivo</i>. Data are representative of six independent experiments. Error bars represent SEM; *<i>P</i><0.05.</p

Apoptosis was increased in the injury model.

<p>(A) Assessment of cell viability by MTT. The cell viability was compared in the different NP cell groups (∗P <0.05). Data are representative of three experiments; error bars represent SEM. (B) Contour diagram of FITC-Annexin V/PI FCM of human NP cells. The graphs represent typical results of cell apoptosis; values represent the means of two experiments. (C) Results of cellular apoptosis was expressed as a fold change. Results are shown as mean ± SEM. Data are representative of three independent experiments (*P<0.05, **P<0.01).</p

The role of PI3K/AKT pathway in miR-27a induced apoptosis.

<p>(A) Western blot analysis of total PI3KCD, p-AKT, NF-κB, Bcl-2 and cleaved caspase 3 protein levels inhuman NP cells with up-regulation and knockdown of miR-27a. (B) Quantification of band intensity in (A). Results are shown as mean ± SEM. Data are representative of three independent experiments (*P<0.05 vs control).</p