Statistical analysis of all cases for Enneking functional score.

<p>Multivariate analysis was performed using seventeen factors of the patient’s background to determine which influence Enneking’s functional score or active range of motion for all cases and for total scapulectomy cases, separately. The amount of remaining bone influenced the Enneking functional score, which means that preserving the glenoid or the acromion lead to better function compared to total scapulectomy. However, there was no significant evidence that reconstruction improved total functional outcome.</p

New classification of scapulectomy.

<p>Five categories are created in terms of resection area. Preserving glenoid or acromion lead to better function compared to total scapulectomy. Preoperative planning with this classification will contribute to expected postoperative function.</p

Summary of multivariate analysis.

<p>Factors that influenced functional outcome include the amount of remaining bone, soft tissue reconstruction, length of the resected humerus and nerve resection. As for total scapulectomy cases, soft tissue reconstruction did not lead to better total functional score but did allow better dexterity of affected hand. This result encourages doing soft tissue reconstruction after scapulectomy, which had been uncertain to improve functional outcome.</p

Patient’s characteristics and functional outcomes.

<p>This retrospective study comprised 48 patients who underwent total or subtotal scapulectomy (more than half of the scapula was resected) and followed for at least one year after surgery. Patients were registered at the Japanese Musculoskeletal Oncology Group affiliated hospitals. Using the Enneking functional score, function and hand position, which reflect shoulder ability, had low scores, but pain and dexterity, which reflect usefulness of the hand joints, had satisfactory scores. The mean total score was 21.1 out of 30 (12–30), which overall is a satisfactory score following resection of the shoulder girdle. Data are expressed as mean±SD.</p

Comparison of soft tissue reconstruction or no reconstruction after total scapulectomy.

<p>As for total scapulectomy cases, soft tissue reconstruction does not improve total functional score but does improve dexterity of the affected hand. Previously there was no clear evidence that soft tissue reconstruction after scapulectomy improved functional outcome.</p

Effects of geldanamycin or onalespib on the PGF_2α-induced expression levels of IL-6 mRNA in MC3T3-E1 cells.

<p>The cultured cells were pretreated with 0.3 μM of geldanamycin (A), onalespib (B) or vehicle for 60 min, and then stimulated by 10 μM of PGF_2α or vehicle for 3 h. The respective total RNA was then isolated and transcribed into cDNA. The expressions of IL-6 mRNA and GAPDH mRNA were quantified by RT-qPCR. The IL-6 mRNA levels were normalized to those of GAPDH mRNA. Each value represents the mean ± S.E.M. of triplicate determinations from three independent cell preparations. ^*<i>p</i><0.05 compared to the value of control. ^**<i>p</i><0.05 compared to the value of PGF_2α alone.</p

Schematic illustration of the regulatory mechanism underlying the amplifying effect of HSP90 inhibitors on the PGF_2α-induced IL-6 synthesis in osteoblast-like MC3T3-E1 cells.

<p>PGF_2α, prostaglandin F_2α; MAPK, mitogen-activated protein kinase; HSP90, heat shock protein 90; IL-6, interleukin-6.</p

Effect of PGF_2α on the phosphorylation of p38 MAP kinase in HSP90 knockdown MC3T3-E1 cells.

<p>(A) The cultured cells were transfected with 10 nM negative control siRNA (Neg) or 10 nM HSP90-siRNA (#1). (B) The cultured cells were transfected with 30 nM negative control siRNA (Neg) or 30 nM HSP90-siRNA (#2). Twenty-four hours after transfection, the cells were stimulated by 10 μM PGF_2α or vehicle for 10 min. The cell extracts were then subjected to SDS-PAGE with subsequent Western blot analysis with antibodies against phospho-specific p38 MAP kinase, p38 MAP kinase, HSP90 or GAPDH. (a) The histogram shows the quantitative representations of the levels of phosphorylated p38 MAP kinase after normalization with respect to GAPDH obtained from laser densitometric analysis. The levels were expressed as the fold increase to the basal levels presented as lane 1. (b),(c) The histogram shows the quantitative representations of the levels of (b) total p38 MAP kinase and (c) HSP90αβ after normalization with respect to GAPDH obtained from laser densitometric analysis, respectively. The levels were expressed as the ratio to the levels presented as lane 1. Each value represents the mean ± S.E.M. of triplicate determinations from three independent cell preparations. ^*<i>p</i><0.05 compared to the value of control. ^**<i>p</i><0.05 compared to the value of PGF_2α alone.</p

Effect of SB203580 on the enhancement by geldanamycin, 17-AAG or 17-DMAG of the PGF_2α-induced IL-6 release in MC3T3-E1 cells.

<p>The cultured cells were preincubated with 30 μM of SB203580 or vehicle for 60 min, subsequently pretreated with 1 μM of geldanamycin, 1 μM of 17-AAG, 1 μM of 17-DMAG or vehicle for 60 min, and then stimulated by 10 μM of PGF_2α or vehicle for 48 h. IL-6 concentrations of the conditioned mediums were determined by ELISA. Each value represents the mean ± S.E.M. of triplicate determinations from three independent cell preparations. ^*<i>p</i><0.05 compared to the value of control. ^**<i>p</i><0.05 compared to the value of PGF_2α alone. ^***<i>p</i><0.05 compared to the value of PGF_2α with the pretreatment of each HSP90 inhibitor.</p

Effects of geldanamycin or 17-AAG on the phosphorylation of MYPT-1, the amounts of MYPT-1, RhoA and Rho-kinase induced by PGF_2α in MC3T3-E1 cells.

<p>The cultured cells were pretreated with various doses of geldanamycin (A) or 17-AAG (B) for 60 min, and then stimulated by 10 μM of PGF_2α or vehicle for 2 min. The cell extracts were then subjected to SDS-PAGE with subsequent Western blot analysis with antibodies against phospho-specific MYPT-1, MYPT-1, RhoA or Rho-kinase. (a) The histogram shows the quantitative representations of the levels of phosphorylated MYPT-1 after normalization with respect to MYPT-1 obtained from laser densitometric analysis. The levels were expressed as the fold increase to the basal levels presented as lane 1. (b),(c),(d) The histogram shows the quantitative representations of the levels of MYPT-1 (b), RhoA (c) and Rho-kinase (d) after normalization with respect to GAPDH obtained from laser densitometric analysis, respectively. The levels were expressed as the ratio to the levels presented as lane 1. Each value represents the mean ± S.E.M. of triplicate determinations from three independent cell preparations. ^*<i>p</i><0.05 compared to the value of control. N.S. designates no significant difference between the indicated pairs.</p