Analysis of protein-protein interactions in cross-talk pathways reveals CRKL protein as a novel prognostic marker in hepatocellular carcinoma.

Deciphering the network of signaling pathways in cancer via protein-protein interactions (PPIs) at the cellular level is a promising approach but remains incomplete. We used an in situ proximity ligation assay to identify and quantify 67 endogenous PPIs among 21 interlinked pathways in two hepatocellular carcinoma (HCC) cells, Huh7 (minimally migratory cells) and Mahlavu (highly migratory cells). We then applied a differential network biology analysis and determined that the novel interaction, CRKL-FLT1, has a high centrality ranking, and the expression of this interaction is strongly correlated with the migratory ability of HCC and other cancer cell lines. Knockdown of CRKL and FLT1 in HCC cells leads to a decrease in cell migration via ERK signaling and the epithelial-mesenchymal transition process. Our immunohistochemical analysis shows high expression levels of the CRKL and CRKL-FLT1 pair that strongly correlate with reduced disease-free and overall survival in HCC patient samples, and a multivariate analysis further established CRKL and the CRKL-FLT1 as novel prognosis markers. This study demonstrated that functional exploration of a disease network with interlinked pathways via PPIs can be used to discover novel biomarkers.

Voice Range Change After Injection Laryngoplasty for Unilateral Vocal Fold Paralysis.

OBJECTIVES: Patients with unilateral vocal fold paralysis (UVFP) caused by nerve injury manifest with voice changes. This study investigated vocal performance measured by voice range profile (VRP) in patients with UVFP and changes in VRP in response to intracordal hyaluronate injection. METHODS: Eighty-five patients with UVFP were enrolled prospectively, among whom 68 received intracordal hyaluronate injections. The outcome measurements included VRP, acoustic and aerodynamic analyses, peak turn frequency of thyroarytenoid-lateral cricoarytenoid muscle complex (TA-LCA) measured by laryngeal electromyography, and normalized glottal gap area by videolaryngostroboscopy. RESULTS: The peak turn frequency of the paralyzed TA-LCA showed a modest correlation with max fundamental frequency (F0) and F0 range. Closed-phase normalized glottal gap area showed modest negative correlations with max F0 and F0 semitone range. Regarding conventional acoustic and aerodynamic analyses, the paralyzed TA-LCA peak turn frequency was only correlated with maximal phonation time. Intracordal hyaluronate injection improved VRP performance by increasing max F0, decreasing min F0, increasing F0 range, and increasing semitone range (all P <0.01) with small or medium strength of effect size (Cohen d, 0.39-0.76). CONCLUSIONS: Change in voice pitch in patients with UVFP can partly predict impairment of neuromuscular functions and glottal gap. VRP provides a more sensitive reflection of the severity of neuromuscular impairment, compared with conventional voice analysis. The validity of VRP is further supported by a robust response to voice improvements following injection laryngoplasty.

Implications of Synkinesis in Unilateral Vocal Fold Paralysis.

Objectives In patients with unilateral vocal fold paralysis (UVFP), laryngeal electromyography (LEMG) occasionally observes synkinesis in laryngeal muscles, a condition that could impair vocal fold mobility and voice control. This study aims to evaluate the impact of synkinesis on UVFP patients. Study Design A retrospective case-control study. Setting Medical center. Subjects and Methods Patients with UVFP onset >6 months were recruited (N = 104). The outcome measurements included LEMG, quantitative LEMG analysis of thyroarytenoid-lateral cricoarytenoid (TA-LCA) muscle complex, glottal gap measured by videolaryngostroboscopy, voice-related quality of life, and voice acoustic analysis. Results According to the LEMG analysis, 8 patients (8%) had synkinesis, and 96 (92%) did not. In the synkinesis group, TA-LCA turn frequency in the lesioned side was comparable to that in the healthy side ( P = .52). Patients in the synkinesis group had higher TA-LCA turn frequency ( P = .001), higher probability of cricothyroid muscle dysfunction ( P = .04), and better voice-related quality of life ( P = .01) but objective voice outcomes comparable to those in the nonsynkinesis group. Conclusions Patients with synkinesis will have near-complete restoration in TA-LCA turn frequency but still experience voice impairment, a finding that is compatible with the mechanism of aberrant reinnervation. However, patients with synkinesis have better disease-related quality of life than do those without synkinesis.