FIP-nha, a fungal immunomodulatory protein from Nectria haematococca, induces apoptosis and autophagy in human gastric cancer cells via blocking the EGFR-mediated STAT3/Akt signaling pathway.

FIP-nha, a fungal immunomodulatory protein from Nectria haematococca, has been demonstrated a broad spectrum of antitumor activity and cell selectivity against human cancers in our previous study. However, the effect and mechanism of FIP-nha on gastric cancer remains unclear. In this study, we systematically observed the cytotoxicity, biological effect, regulatory mechanism and interaction target of FIP-nha on human gastric cancer cell lines, AGS and SGC7901. Our results demonstrated that FIP-nha inhibited the growth of AGS and SGC7901 cells in a dose-dependent manner and exerted proapoptotic effects on both cells as confirmed by flow cytometry, DAPI staining and western blot analysis. Additionally, the exposure of AGS and SGC7901 to FIP-nha induced autophagy as indicated by western blot analysis, GFP-LC3 and mCherry-GFP-LC3 transfection and acridine orange staining. Furthermore, we found that FIP-nha decreased the phosphorylation of EGFR, STAT3 and Akt and inhibited activation effect of ligand factor EGF to EGFR and its downstream signal molecule STAT3 and Akt. Finally, we proved that FIP-nha located on the surface of gastric cancer cells and bound directly to the transmembrane protein of EGFR by immunoprecipitation, cellular localization, molecular docking, microscale thermophoresis assay. The above findings indicated that FIP-nha inhibited the growth of gastric cancer and induced apoptosis and autophagy through competitively binding to EGFR with EGF to blocking the EGFR-mediated STAT3/Akt pathway. In summary, our study provided novel insights regarding the activity of FIP-nha against gastric cancer and contributed to the clinical application of FIP-nha as a potential chemotherapy drugs that targeted EGFR for human gastric cancer.

Induction chemotherapy for the individualised treatment of hypopharyngeal carcinoma with cervical oesophageal invasion: a retrospective cohort study.

BACKGROUND: This study aimed to evaluate the potential of induction chemotherapy as an indicator of the management of advanced hypopharyngeal carcinoma with cervical oesophageal invasion. METHODS: Sixty-eight patients admitted to our hospital between February 2003 and November 2016 with stage IVB hypopharyngeal carcinoma with cervical oesophageal invasion were retrospectively analysed. Patients were divided into two groups according to the treatment they selected following an explanation of the different treatments available. Patients in group A received induction chemotherapy and had (1) complete/partial remission following chemotherapy and radiotherapy/concurrent chemoradiotherapy or (2) stable disease following chemotherapy and surgery. Patients in group B underwent surgery followed by adjuvant radiotherapy/concurrent chemoradiotherapy. Survival analyses were performed using the Kaplan-Meier method, and differences between the groups were evaluated using the log-rank test. Laryngeal and oesophageal retention rates were compared using the cross-tabulation test. RESULTS: The 3- and 5-year overall survival rates were 22.86% and 11.43% in group A and 24.25% and 6.06% in group B, respectively (all P > 0.05). The laryngeal and oesophageal retention rates were 40.0% and 74.3% in group A and 0.0% and 27.3% in group B, respectively (all P < 0.01). There was no statistically significant difference in the incidence of post-operative complications between the two groups (group A 8.6%, group B 12.1%; P > 0.05). CONCLUSIONS: Induction chemotherapy may be an appropriate first choice to ensure laryngeal and oesophageal preservation in the individualised treatment of advanced hypopharyngeal carcinoma with cervical oesophageal invasion.

Silencing novel long non-coding RNA FKBP9P1 represses malignant progression and inhibits PI3K/AKT signaling of head and neck squamous cell carcinoma in vitro.

BACKGROUND: Long non-coding RNAs (lncRNAs) play key roles in human cancers. In our previous study, we demonstrated that lncRNA FKBP prolyl isomerase 9 pseudogene 1 (FKBP9P1) was highly expressed in head and neck squamous cell cancer (HNSCC) tissues. However, its functional significance remains poorly understood. In the present study, we identify the role and potential molecular biologic mechanisms of FKBP9P1 in HNSCC. METHODS: Quantitative real-time polymerase chain reaction was used to detect the expression of FKBP9P1 in HNSCC tissues, matched adjacent normal tissues, human HNSCC cells (FaDu, Cal-27, SCC4, and SCC9), and human immortalized keratinocytes cell HaCaT (normal control). Cal-27 and SCC9 cells were transfected with sh-FKBP9P1-1, sh-FKBP9P1-2, and normal control (sh-NC) lentivirus. Cell counting kit-8 assay, colony formation assay, wound healing assay, and trans-well assay were used to explore the biologic function of FKBP9P1 in HNSCC cells. Furthermore, western blotting was used to determine the mechanism of FKBP9P1 in HNSCC progression. Chi-squared test was performed to assess the clinical significance among FKBP9P1 high-expression and low-expression groups. Survival analyses were performed using the Kaplan-Meier method and assessed using the log-rank test. The comparison between two groups was analyzed by Student t test, and comparisons among multiple samples were performed by one-way analysis of variance and a Bonferroni post hoc test. RESULTS: FKBP9P1 expression was significantly up-regulated in HNSCC tissues (tumor vs. normal, 1.914 vs. 0.957, t = 7.746, P < 0.001) and cell lines (P < 0.01 in all HNSCC cell lines). Besides, the median FKBP9P1 expression of HNSCC tissues (1.677) was considered as the threshold. High FKBP9P1 level was correlated with advanced T stage (P = 0.022), advanced N stage (P = 0.036), advanced clinical stage (P = 0.018), and poor prognosis of HNSCC patients (overall survival, P = 0.002 and disease-free survival, P < 0.001). Knockdown of FKBP9P1 led to marked repression in proliferation, migration, and invasion of HNSCC cells in vitro (P all < 0.01). Mechanistically, silencing FKBP9P1 was observed to restrain the PI3K/AKT signaling pathway. CONCLUSIONS: Silencing lncRNA FKBP9P1 represses HNSCC progression and inhibits PI3K/AKT (phosphatidylinositol 3 kinase/AKT Serine/Threonine Kinase) signaling in vitro. Therefore, FKBP9P1 could be a potential new target for the diagnosis and treatment of HNSCC patients.

[Study on the stability of perceptual evaluation of voice quality].

OBJECTIVE: To explore the factors that influence the stability of evaluation results judged by a jury through a standard research on perceptual evaluation measurements of voice quality. METHODS: Voice samples from 300 patients with dysphonia and 100 control subjects with normal voice were recorded and assessed by a jury composed of 6 experienced listeners from different hospitals. The voice samples were discourse voices and ordered randomly 3 times, and the mean of 3 evaluations using visual analogue scale were the final results. The jury was instructed to classify voice samples according to the G (grade), R (rough) and B (breathy) components of the GRBAS scale on a 4-point scale ranging from 0 for normal to 3 for severe dysphonia. Κ value was used to analyze the concordance of evaluation results and regression analysis was used to research the effects of the extent of voice disorder to the stability of perceptual evaluation. RESULTS: The discordance of evaluation existed both between the jury and in listeners themselves. The concordance of listeners themselves of each evaluation parameter was not bad, good, or even very good, and the concordance of evaluation of G was the best (κ value: 0.46 - 0.85), then R (κ value: 0.41 - 0.84) and B (κ value: 0.41 - 0.81). The concordance between the jury was worse than that in themselves. And except a listener whose concordance of evaluation was under the requirement, the concordance of evaluation of G was the best (κ value: 0.43 - 0.96), then R (κ value: 0.33 - 0.78) and B (κ value: 0.002 - 0.45). The stability of evaluation of normal voice and severe voice disorder was better than mild and moderate voice disorder. CONCLUSIONS: The discordance between the jury was the main factor that influence the stability of perceptual evaluation. The evaluation parameters and extent of voice disorder will influence the stability of perceptual evaluation of the jury.

[Self-assessment characteristics of voice handicap index for voice disorders and its influencing factors].

OBJECTIVE: To investigate the self-assessment characteristics of Voice Handicap Index (VHI) for voice disorders and its influencing factors. METHODS: One thousand seven hundred and sixty six dysphonic patients and 120 control subjects were included in this study. Two hundred twenty seven patients were treated with phonosurgery or Botulinumtoxin injection. VHI was used for self-assessment. RESULTS: Dysphonic patients had worse VHI scores than control (z from 8.039 to 17.043, P = 0.000). There was a significant difference among the VHI scores of different diseases. VHI scores were descending in order among spasmodic dysphonia, vocal fold paralysis, functional dysphonia, sulcus vocalis, benign and malignant tumor of vocal fold, vocal fold cyst, Reinke's edema, vocal fold polyp, vocal fold keratosis and chronic laryngitis, vocal nodule. The emotional scores were the highest in spasmodic dysphonia, and followed by functional dysphonia. In another group, the physical scores were higher than functional scores and emotional scores. Treatment resulted in statistical improvement in VHI scores (P < 0.05). The total scores were different significantly between different educational background and age groups (F from 8.701 to 27.371, P = 0.000). The higher the educational degree, the higher the VHI scores. As to age groups, the juvenile group's scores were the lowest, while the youth's group the highest, then the scores declined when ages increased. CONCLUSION: As a useful supplementary instrument to measure the voice disorder severity and the treatment's effect, VHI can comprehensively assess the voice handicap's affect to the life quality and the difference after the treatment, especially in physical, functional and emotional aspects, but it is somehow subject to the educational degree and age.

[Analysis of reliability and validity of the Chinese version of voice handicap index (VHI)].

OBJECTIVE: To investigate the reliability and validity of the Chinese version of voice handicap index (VHI). METHODS: The cross-cultural adaptation of health-related quality of life (HRQOL) measures was used to evaluate the Chinese version of VHI. Five hundred forty six dysphonic patients and 80 control subjects were included, 30 of patients and 20 of control subjects also had Hong Kong version VHI test simultaneously. RESULTS: The internal consistency reliability of overall VHI scores and three subscale scores of the Chinese version of VHI were 0.8657-0.9517. The reliability coefficients (test to retest, 2-week interval) was 0.992 (P < 0.001). The correlation coefficient of overall VHI scores and three subscale scores and internal subscale scores were 0.643-0.904 (P < 0.01). There were no significant difference between the Chinese version and Hong Kong Chinese version (Z = 0.397, P = 0.691 ) with high dependability (r = 0.995, P < 0.001). The factor analysis of construct validity shows that the eigenvalue of 6 factors is above 1. The cumulative proportion was 77.24%. The loading was higher than 0.4 among every item. VHI total scores were significantly higher in dysphonic patients than in control subjects (Z = 17.69, P = 0.000). This is also true for all VHI subscores in the functional (Z = 14.14, P = 0.000), physical (Z = 17.68, P = 0.000) and emotional domains (Z = 15.50, P = 0.000). CONCLUSIONS: The Chinese version of VHI had a good reliability and validity. It can be used to evaluate dysphonic patients.

[Use of endoscopic placement of laryngeal stent combined with mucosal suturing of vocal fold for the treatment of laryngeal web].

OBJECTIVE: To investigate the effects of laryngeal stent combined with vocal fold mucosal suture for treating laryngeal web. METHODS: A retrospective study performed, 21 laryngeal web cases were included in this study. The web causes were previous vocal fold surgery for benign lesions in 8 cases, previous laryngeal surgery for recurrent papilloma in 6 cases, laryngeal trauma in 6 cases, and congenital web in 1 case. Fifteen adults received glottic silastic sheet keels placement with vocal fold suture, another 2 adults and 4 children only had vocal fold suture, the above procedures were performed with laryngeal endoscopy. RESULTS: Among all patients, good glottis shape was achieved and good voice maintained, 6 months to 3 years follow-up showed open airway without restenosis. 15 adults who had both keels placement and vocal cord suture tolerated the keel well. Keel removal under direct laryngoscopy was performed 3-4 weeks after surgery. A normal triangular shaped glottis anterior commissure anatomy was restored in 14 patients and 2-3 mm residual webbing was found in 1 patient (who had laryngofissure history). 2-3 mm residual webbing was found in 4 children who only had endoscopic vocal fold suture. After receiving endoscopic vocal fold suture, 2 adults who was found to have 2-3 mm normal mucosal membrane in anterior commissure during surgery, restored normal vocal cord shape. CONCLUSIONS: The study shows that silastic keel placement combined with vocal fold mucosal suture was successful for treating laryngeal webs cases. Mucosal suturing of vocal fold may decrease or eliminate raw surface exposure and it is useful in the management of pediatric laryngeal webs and webs with normal mucosal membrane in anterior commissure. The potential benefits of those procedures are in avoiding long-term glottic stenting, laryngofissure, and tracheotomy.

[Characteristics of normal laryngeal nerve evoked potentials].

OBJECTIVE: To study the behaviors of normal recurrent laryngeal nerve and superior laryngeal nerve evoked electromyography, to know the reference values of related parameters. METHODS: Thirty six normal subjects were studied with laryngeal evoked electromyography. The latent period, duration, and amplitude of the evoked potentials (EP) recorded in laryngeal muscles were investigated. RESULTS: The latent period of the EP recording from the thyroarytenoid muscles (TA), after stimulating recurrent laryngeal nerve (RLN), was (1.13 - 2.25) ms (x +/- 2s), that of TA after stimulating vagus nerve (VN) was (2.57 - 5.85) ms. There were significant differences in the latent periods of EP between RLN and VN (P < 0.01), and the latent periods of EP recording from TA after stimulating right cervical part of VN, (2.01 - 4. 53) ms (x +/- 2s) were significant shorter than those of left, (3.70 - 6.98) ms (x +/- 2s, P < 0.01). The durations of the EP were somewhat different but not significant. The amplitudes of the EP in different laryngeal muscles changes markedly. The wave form and latent period of the EP recording from cricothyroid muscle (CT), after stimulating the external branch of superior laryngeal nerve (SLN), was similar with the EP of RLN, the reflective EP R1 and R2 could be recorded from TA, posterior cricoarytenoid muscles (PCA), CT after stimulating the internal branch of SLN. CONCLUSIONS: The characteristics of the EP recorded in laryngeal muscles after stimulation of RLN, SLN and VN are different. The latent period, duration and amplitude are important parameters in the evaluation of laryngeal nerves function, the latent period is the most important parameter.

[Laryngopharyngeal characteristics of myasthenia gravis].

OBJECTIVE: To observe the laryngopharynx manifestation and electromyography characteristics of myasthenia gravis (MG) patients. METHODS: Thirty cases of MG were included in this study, their laryngopharynx symptoms and signs, voice acoustic assessment, laryngeal electromyography (LEMG) behaviors and repetitive nerve stimulation test(RNS) were analyzed, and the data was compared with that of normal subjects. RESULTS: About 36.7% of MG patients (11/30) had the symptoms of hoarseness, voice fatigue, dysphonia and dysphagia. The vocal folds movements of 16.7% of MG patients(5/30) appeared weaker than normal, and their vocal glottic couldn't close completely, while with a seam during phonation. Voice amplitude (68.3 +/- 14.6) dB (x +/- s, same at below), and maximum phonation time (15.1 +/- 4.0) s, were greatly lower than normal; shimmer(2. 43 +/- 1.19)%, and normalized noise energy (-9.6 +/- 3.3) dB, were greatly higher than normal. The amplitudes of interference patterns in MG patients' LEMG markedly decreased, except introarytenoid muscle, during low, normal and high pitch phonation, the amplitudes of thyoiarytenoid muscle were (215 +/- 69) microV, (298 +/- 113) microV and (380 +/- 153) microV, those of cricoarytenoid muscle were (253 +/- 92) microV, (361 +/- 116) microV and (486 +/- 155) RV. The turns increased but had no statistical difference. In the RNS test, 83.3% MG patients (25/30) showed masculine response. There were about 2.20 +/- 1.32 pieces of laryngeal muscles involved, and the reduction rate in amplitude of the compound muscle action potential for RNS was about (27.9 +/- 19.2)%. CONCLUSIONS: Only parts of MG patients had laryngopharyngeal symptoms, but the laryngeal muscles of most of them were involved, appearing as the masculine response for RNS, the decreased synchronization of the laryngeal muscles' interference patterns, the decreased capacity of phonation. MG must be differentiated when a patient has the symptoms of voice weakness, hoarseness and dysphonia. Laryngeal RNS test should be used in the early diagnosis of MG.

[Laryngeal electromyographic characteristics of vocal fold immobility].

OBJECTIVE: To investigate the characteristics of laryngeal electromyography and evoked electromyography in vocal fold immobility. METHODS: 108 cases of vocal fold immobility were analyzed by clinical manifestation and laryngeal electromyography characteristics, including spontaneous potential activity, motor unit potential (MUP) measurement, recruitment pattern analysis and evoked electromyography evaluation. RESULTS: Neurogenic vocal fold immobility showed a wide variety of abnormal activity. Fibrillation potentials and positive sharp waves were found in patients with laryngeal nerve injuries. For laryngeal paralysis, there was no reaction with LEMG and evoked LEMG. For laryngeal paresis, it showed decreased MUP and decreased recruitment activities with simple pattern recruitment or mixed pattern recruitment, and decreased evoked muscle response potentials were also shown with delayed latency than normal (P < 0.05) in the TA (2.2 +/- 1.0) ms(x +/- s), PCA (2.4 +/- 1.0) ms and lower amplitude in the TA (0.9 +/- 0.7) mV and PCA (1.2 +/- 1.0) mV (P < 0.05). Patients with vocal fold mechanical limitations generally yielded normal LEMG and Evoked LEMG. Patients with neoplastic infiltration of the laryngeal muscles demonstrated decreased LEMG and nearly normal Evoked LEMG with normal latency and lower amplitude. CONCLUSIONS: General EMG and evoked EMG play a crucial role in the diagnosis of vocal fold immobility.

[Patterns of spasmodic dysphonia and botulinum toxin injections].

OBJECTIVE: To investigate the patterns of spasmodic dysphonia and the outcome treated with botulinum toxin A injections. METHODS: All subjects were studied with acoustic analysis, laryngostroboscopy and laryngeal electromyography (EMG) including motor unit potential measure (MUP), recruitment pattern analysis and evoked electromyography. All the patients with spasmodic dysphonia were received botulinum toxin A (BOTOX) injections in each affected muscles and mostly under electromyographic guidance. RESULTS: Among 22 cases of spasmodic dysphonia, 18 cases of adductor dysphonic patients have strained, strangled voice with intermittent breaks in speech as a consequence of hyperadduction and spasm of the vocal folds during phonation. Two patients had synchronous pharyngeal, lingual and velar tremor. Amplitudes of MUP of thyroarytenoid muscle (TA) were greater in patients group than in normal group (P < 0.01); The recruitment activity was increased and the amplitudes were greater than normal group (700-2500 microV) and the duration of activity of the TA during phonation was also notably greater in patients group than in normal group. Four cases of abductor dysphonic patients have a breathy, effortful hypophonic voice with abrupt termination of voicing. Amplitudes of MUP of posterior cricoarytenoid muscle (PCA) in patients group were increased up to 374 to 538 microV. The recruitment activity was increased and the amplitude was greater than normal(3000-5000 microV). In the adductor dysphonic group, patients who were treated with unilateral toxin injection had good results with 2.5 U or more. The average onset of toxin effect in all adductor dysphonic patients was at 6 hours to 2 days (1.4+/-0. 8) days (x +/- s), with a peak effect at 2 weeks and the follow-up EMG showed fibrillation potentials or electric silence in injected muscle. Duration of benefit was 8 to 24 weeks (15.2 +/- 4.9) weeks. The side-effect of toxin injection were including breathy voice or occasional dysphagia and aspiration. The patients with abductor spasms were less well controlled after PCA injections. CONCLUSIONS: Spasmodic dysphonia was regarded as a neuromuscular diseases, so its diagnosis, classification, treatment and follow-up should depend on not only clinical manifestation but also EMG. Presently, for controlling the dystonic symptoms, the most effective therapy for most of those patients is local BOTOX injections. Repeated injections are required to have a stable results.