Combined treatment with acetazolamide and cisplatin enhances the chemosensitivity of human head and neck squamous cell carcinoma TU868 cells.

AIMS: To investigate whether combination of acetazolamide and cisplatin can enhance the chemosensitivity of human head and neck squamous cell carcinoma (HNSCC) cell line TU868. METHODS: MTT assay was performed to determine the effect of acetazolamide, cisplatin and their combination on the proliferation of TU868 cells. Then the effect of these 2 drugs on the expression of proliferation-related and apoptosis-related proteins was detected by Western blot. Moreover, the effect of acetazolamide and cisplatin on the expression of aquaporin-1 was detected by RT-qPCR. Loss-of-function assays was performed to assess whether the effect of acetazolamide and cisplatin on TU868 cells was mediated by aquaporin-1. The effect of acetazolamide and cisplatin on tumor cell growth was confirmed in mice by testing the tumor growth size. RESULTS: Acetazolamide and cisplatin treatment displayed synergistic effects on the inhibition of TU868 cell growth compared with the drugs used alone. Moreover, the acetazolamide/cisplatin combination could decrease the level of PCNA but increase the level of p53; decrease the ratio of Bcl-2/Bax and increase the expression of caspase-3 compared with the single drug treated group. Moreover, we found that the combination also significantly inhibits aquaporin-1 expression. Loss-of-function assays suggested that the anti-tumor effect of these 2 drugs was achieved via affecting aquaporin-1. Consistent with the in vitro assays, combined treatment with acetazolamide and cisplatin significantly inhibits the tumor growth in mice compared with the single drug treated group. CONCLUSION: These results demonstrated that combined treatment with acetazolamide and cisplatin could synergistically inhibit the malignant development of HNSCC cells.

Histiocytic sarcoma of the neck: A case report.

Histiocytic sarcoma (HS) is a term used to describe malignant hyperplasia of cells exhibiting morphological and immunophenotypical characteristics similar to those of mature cells, with expression of one or more tissue cell markers, excluding acute monocytic leukemia and primitive monocytic sarcoma. We herein describe a case of histiocytic sarcoma of the neck supported by histopathological and immunohistological evidence. A 53-year-old female patient of Chinese descent presented with a rapidly enlarging right neck mass. Imaging studies revealed multiple right cervical lymphadenectases with right jugular vein involvement. The tumor was composed of diffusely distributed large non-cohesive tumor cells, round or oval and focally spindle-shaped. The tumor cells were immunopositive for macrophage-associated antigen CD68 and lysosomes, mostly consistent with a diagnosis of HS. HS is very prone to systemic metastasis; therefore, early diagnosis and timely treatment are crucial.

Effects of the Akt inhibitor Src-homology 5 on proliferation and apoptosis of the laryngeal squamous cell carcinoma.

OBJECTIVE: The aim of this study was to investigate the effect of the Akt inhibitor Src-homology 5 (SH-5) on the proliferation and apoptosis of laryngeal squamous cell carcinoma cells (LSCC; Hep-2 cells) and to elucidate the possible mechanisms of such effects. MATERIALS AND METHODS: Hep-2 cells were treated with different concentrations of the Akt inhibitor SH-5. The inhibitory effect of SH-5 on cell proliferation was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, whereas apoptosis was detected by flow cytometric based on Annexin V/propidium iodide (PI) staining. In addition, the expression level of Akt protein was evaluated by Western blot analysis. RESULTS: MTT assay results revealed that SH-5 inhibited the proliferation of Hep-2 cells, with its greatest effect being observed at 2 µM. Apoptosis of Hep-2 cells increased following treatment with SH-5. Treatment of Hep-2 cells with SH-5 decreased the expression of Akt, and this effect was statistically significantly when compared with that in controls (P < 0.05). CONCLUSION: SH-5 inhibited proliferation and induced apoptosis in the LSCC cell line Hep-2. These effects may be caused by inhibition of the phosphoinositide 3-kinase-Akt signaling pathway. We believe that our data will provide useful insights into LSCC target treatment and future researchn.

Bone Age and Serum Osteocalcin Levels in Children With Obstructive Sleep Apnea Hypopnea Syndrome Before and After Adenotonsillectomy.

Our aim was to study the changes in bone age and serum osteocalcin levels before and after adenotonsillectomy (AT) in children with obstructive sleep apnea hypopnea syndrome (OSAHS). A total of 58 OSAHS children (37 males and 21 females) with the mean age of 6.68 ± 1.11 years were enrolled and assessed by x-ray-based bone age estimation and enzyme-linked immunosorbent assay-based measurement of serum osteocalcin levels, before surgery and 6 months after AT. SPSS 19.0 software was used for statistical analysis. Our results revealed that bone age and serum osteocalcin levels in OSAHS patients were significantly lower than normal controls before AT (P < 0.05). Within 6 months after surgery, the bone age and the serum osteocalcin levels were significantly elevated in OSAHS patients (P < 0.05), compared with those before surgery. Serum osteocalcin levels and bone age are negatively correlated with apnea-hypopnea index, oxygen desaturation index, the percentage of the total recorded time spent below 90% oxygen saturation, and Epworth sleepiness scale scores (all P < 0.05). Our findings suggested that bone age and serum osteocalcin levels may be correlated with the development of OSAHS in children. AT may improve bone age and serum osteocalcin levels in OSAHS children.

Salicylate-induced changes in immediate-early genes in the hippocampal CA1 area.

Studies have suggested that salicylate affects neuronal function via interactions with specific membrane channels/receptors. However, the effect of salicylate on activity and synaptic morphology of the hippocampal Cornu Ammonis (CA) 1 area remains to be elucidated. The activation of immediate-early genes (IEGs) was reported to correlate with neuronal activity, in particular activity-regulated cytoskeleton-associated protein and early growth response gene 1. The aim of the present study was to evaluate the expression of these IEGs, as well that of N-methyl D-aspartate (NMDA) receptor subunit 2B in rats following acute and chronic salicylate treatment. Protein and messenger RNA levels of all three genes were increased in rats following chronic administration of salicylate (300 mg/kg for 10 days), returning to baseline levels 14 days post-cessation of treatment. The transient upregulation of gene expression following treatment was accompanied by ultrastructural alterations in hippocampal CA1 area synapses. An increase in synaptic interface curvature was observed as well as an increased number of presynaptic vesicles; in addition, postsynaptic densities thickened and lengthened. In conclusion, the results of the present study indicated that chronic exposure to salicylate may lead to structural alteration of hippocampal CA1 neurons, and it was suggested that this process occurs through induced expression of IEGs via NMDA receptor activation.

[Knowledge and attitude of clinicians to pediatric obstructive sleep apnea hypopnea syndrome].

OBJECTIVE: To investigate the knowledge and attitude of clinicians in the departments of pediatrics and otolaryngology to pediatric obstructive sleep apnea hypopnea syndrome (OSAHS), since in China, the clinicians in these two departments had closest relationship with the diagnosis and treatment of OSAHS in children. METHODS: A validated questionnaire from USA which was the obstructive sleep apnea knowledge and attitudes questionnaire in children (OSAKA-KIDS) was used and permission by original author. The questionnaire was mailed to ENT doctors and pediatricians in 43 public hospitals in Shandong province. RESULTS: OSA-KIDS in Chinese version was re-tested by 30 physicians, r = 0.92. Totally, 391 valid questionnaires (87.7%) were returned. Average of correct rate (x(-) ± s) in 18 knowledge items was 64.1% ± 19.1%. Cronbach's α coefficient was 0.76. There was no difference between ENT doctors and pediatrics in total knowledge score. However, there was significant difference in below 2 questions: ENT doctors had more correction in answer "nearly 2% of children have OSAHS" and pediatrics had more correction in answer "pediatric OSAHS may be associated with pulmonary hypertension". Only 24.3% clinicians correctly know the degree of snoring (mild to severe) was not correlated with the severity of obstructive apnea in children. Only 16.1% could correctly answer the question about cardio-respiratory monitor could not reliably detect both central and obstructive apnea in infant. Cronbach's α coefficient was 0.72 in 5 items which was about importance of disease and self-evaluation in confidence. While more than 90% clinicians stated that "As a clinical disorder OSAHS is important or very, extremely important". However, among them, only about 36% felt confident in identifying or managing children with OSAHS. Total knowledge score about OSAHS was not different by gender or specialty (P > 0.05), but more knowledge was associated with more positive attitudes overall (P < 0.05) and more elder in age or longer years in practice (r = 0.384, P < 0.0001). CONCLUSIONS: It should be paid more effort to elevate the knowledge and attitude about pediatric OSAHS in pediatricians and otolaryngologists.

Identification of epidermal growth factor receptor (EGFR) exon 20 single nucleotide polymorphism in Chinese squamous cell carcinoma of head and neck (SCCHN).

CONCLUSION: The 78th single nucleotide polymorphism (SNP) in exon 20 of epidermal growth factor receptor (EGFR) might be a predictor of the clinical course of squamous cell carcinoma of the head and neck (SCCHN), and genetic predisposition could influence the progression of SCCHN in Chinese subjects. OBJECTIVE: To analyze the clinical significance of EGFR SNP in Chinese SCCHN patients. MATERIALS AND METHODS: Direct sequencing of exons 18-21 was used to analyze somatic mutations of EGFR. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to evaluate the expression level of EGFR. The chi-square test and Kaplan-Meier method were the main statistical methods used to analyze the correlation of the investigated variables and prognostic significance. RESULTS: In analyzing exons 18-21 of EGFR in 96 patients with SCCHN, only one SNP was found in the 78th site of exon 20 and it mostly existed in specimens coming from the hypopharynx. Further statistical analysis showed that among the clinical or histopathologic parameters, the 78th SNP had a close relationship with earlier stage and more localized primary carcinoma, while at present the analysis did not support the proposition that the SNP was an independent prognostic factor.

[Comprehensive treatment of 352 cases with hypopharyngeal cancer].

OBJECTIVE: To review the experience of different surgical construction methods and comprehensive treatments for hypopharyngeal cancer. METHODS: Three hundred and fifty-two cases (According to UICC 2002 criteria, stage I, 3; II, 31; III, 134; IV, 184) with hypopharyngeal cancer were retrospectively reviewed from 1999 to 2005 in the Department of Otorhinolaryngology of Qilu Hospital of Shandong University, including 321 males and 31 females. The median age was 58 years old, ranged from 26 to 82 years old. All the tumors originated from the pyriform sinus (272), posterior pharyngeal wall (61), postcricoid area (19). There were no distant metastasis. Two hundred and fourty-one cases were surgically treated with laryngeal functions preserved and 111 cases without laryngeal functions preserved. All the patients received modified neck dissection, including both unilateral (247 patients) and bilateral (105 patients). Pharyngoesophageal defect reconstruction methods in cases with laryngeal functions preserved were: direct suture in 137, pectoralis major musculocutaneous flap in 62, split graft in 2, pectoralis major musculocutaneous flap combined with the split graft in 5, stomach pulling-up in 12, colon interposition in 23 patients. While in cases without laryngeal functions preserved the methods includes: direct suture in 54, laryngotracheal flap in 54 patients, pectoralis major musculocutaneous flap in 7, laryngotracheal flap combined with pectoralis major musculocutaneous flap in 8, stomach pulling-up in 22, colon interposition in 3 patients. All patients received radiotherapy postoperatively (dose 55 - 75 Gy). RESULTS: The overall 3 and 5 year survival rates were 65.1% (229/352) and 53.6% (142/265), respectively. For stage I, the 5 year survival rate was 3/3, stage II, 80.6%(25/31), stage III, 65.0% (67/103), stage IV, 36.7% (47/128). The 3 and 5 year survival rates in functionally preserved group were 68.0% (164/241) and 59.7% (114/191), respectively, while in non-functionally preserved group were 58.6% (65/111) and 37.8% (28/74), respectively. The cervical lymph node metastasis was found in 239 sides. Pathologic findings showed that well, moderately and lower differentiated squamous cell carcinomas were 84, 163, 105 cases, respectively. Laryngeal functions (voice, respiration and deglutition) were completely restored in 169 patients and partially restored (voice and deglutition) in 72 patients. CONCLUSIONS: Combined surgery and radiotherapy are the best choice for hypopharyngeal cancer. The continuity of the pharyngoesophagus is restored and the laryngeal function is preserved as far as possible. The preservation of laryngeal function and the laryngeal and pharyngeal reconstruction are based on the premise that the tumor was excised completely.

Therapeutic influence on circulating and monocyte-derived dendritic cells in laryngeal squamous cell carcinoma patients.

CONCLUSION: Surgical resection of laryngeal squamous cell carcinoma (LSCC) could be associated with improved circulating myeloid dendritic cell (mDC) number and monocyte-derived dentritic cell (MoDC) function. Although adjunctive radiotherapy after surgery did not effect the normalization of mDC number, it may have an impact on MoDC function. OBJECTIVE: To investigate the effects of surgery and adjunctive radiotherapy on both circulating dendritic cells (DCs) and MoDCs of LSCC patients. SUBJECTS AND METHODS: Forty-six patients with LSCC and 15 age-matched healthy control subjects were enrolled in this study. Blood samples were taken before, during, and after conventional treatment from both patients who underwent surgery only (n=18) and those who had adjunctive radiotherapy after tumor removal (n=28). Three-color flow cytometry was used for determination of circulating DC subsets. Moreover, MoDCs were generated utilizing granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4), then the phenotype of MoDCs was measured by flow cytometry and the ability to stimulate autologous T cells was tested in a mixed leukocyte reaction (MLR). RESULTS: The preoperative mDC counts, MoDC surface molecular expression and stimulatory capacity were impaired in patients in comparison with controls. The number of mDCs and the expression of CD80, CD83, and HLA-DR on MoDCs were significantly increased as compared with those pretreatment in patients who underwent surgery only and in those who had surgery followed by adjunctive radiotherapy. However, the recovery of CD86 expression and allostimulatory activity was only observed in patients who underwent surgery alone.

[Surgical treatment of hypopharyngeal cancer with cervical esophageal invasion].

OBJECTIVE: To review the experience of different surgical construction methods for hypopharyngeal cancer with cervical esophageal invasion. METHODS: From 1989 to 2000,forty-eight patients with advanced hypopharyngeal cancer and cervical esophageal invasion were retrospectively reviewed, including 38 males and 10 females. The median age was 54. 3 years old, ranged from 26 to 71 years old. According to UICC 1997 criteria, all the tumors were T4 stage and originated from the pyriform sinus (33), posterior pharyngeal wall (14), postcricoid area (1), there were 28 patients in cN0, 15 in cN1, 5 in cN2 and no distant metastasis. Precise preoperative evaluation was performed with computed tomography scan, barium swallow perspective and biopsy. All the patients received modified neck dissection, including both unilateral (38 patients) and bilateral (10 patients). Pharyngoesophageal defect reconstruction methods were: laryngotracheal flap in 11 patients, pectoralis major musculocutaneous flap in 13, laryngotracheal flap combined with pectoralis major musculocutaneous flap in 6, pectoralis major musculocutaneous flap combined with the split graft in 10, stomach pulling-up in 3, colon interposition in 5 patients. Total laryngectomy was carried out in 8 patients. All patients received radiotherapy postoperatively (dose 55 - 75 Gy). RESULTS: The cervical lymph node metastasis was found in 20 patients. Pathologic findings showed that well, moderately and lower differentiated squamous cell carcinomas were 18, 24, 6 cases, respectively. The overall 3 and 5 year survival rates were 52.1% (25/48) and 27.3% (12/44), respectively. The 3 and 5 year survival rates in functionally preserved group were 65.2% (15/23) and 33.3% (7/21), while in non functionally preserved group were 40.0% (10/25) and 21.7% (5/23), respectively. Fifteen patients laryngeal functions (voice, respiration and deglutition) were completely restored and 8 patients partially restored (voice and deglutition). The decannulation rate was 65% (15/23). The complication included pharyngeal fistulas in 10 cases and splitting of chest wall in 1 cases. CONCLUSIONS: Combined therapy was the best choice for hypopharyngeal cancer with cervical esophageal invasion. The laryngeal function is preserved as far as possible. The continuity of the pharyngoesophagus was restored by pectoralis major musculocutaneous flap, laryngotracheal flap, or combined with the split graft. Stomach transposition or colon interposition was used while the defect of the esophagus was greater.

[Validity of high negative pressure drainage for chylous fistula after neck dissection: a report of eight cases].

BACKGROUND & OBJECTIVE: Chylous fistula, a severe complication after operation on neck, has close correlation with definite anatomical position and variation. Its treatment remains controversial. This study was to evaluate the validity of high negative pressure drainage for chylous fistula after neck dissection. METHODS: A treatment of high negative pressure (-30 to -50 kPa) drainage, fasting, and reasonable venous nutrition was applied to 8 patients with postoperative chylous fistula. RESULTS: Of the 8 patients, 7 recovered smoothly without severe complication, and pectoralis major muscle flap was adopted to cure the failed one. CONCLUSION: A treatment of high negative pressure drainage and reasonable diet is effective and safe for chylous fistula at early stage.

[Substitution of esophagus with stomach or colon without thoracotomy in the treatment of cervical esophageal carcinoma with laryngeal function preserved].

OBJECTIVE: To study the feasibility and effect of substituting esophagus with stomach or colon without thoracotomy in the treatment of cervical esophageal carcinoma with laryngeal function preserved. METHODS: Twenty-four patients with cervical esophageal carcinoma were retrospectively reviewed. The esophagus was resected and substituted with 19 gastric pull-up and 5 colon interposition. Nineteen patients received radiotherapy postoperatively (dose 50 - 70 Gy). RESULTS: Twenty two patients were follow up over 3 years. The 3- and 5-year survival rates for T2 were 3 and 1, for T3, T4 8 and 3, respectively. The laryngeal function preservation rate was 77% (17/24) and the decannulation rate was 75% (12/16). The complication rate was 29%. CONCLUSION: Surgical resection of cervical esophageal carcinoma with removal of the extraesophageal invaded tissues while preserving the laryngeal function is possible. The continuity of the esophagus is restored by stomach transposition and colon interposition. Combined with radiotherapy, the survival rate and life quality of the patient might be improved.

[Surgical management of benign pharyngo-esophageal stenosis].

OBJECTIVE: To study the methods and outcome of surgical management for pharyngo-esophageal stenosis. METHODS: Twenty-seven patients with pharyngo-esophageal stenosis from January 1983 to June 2001 were reviewed. Among 27 cases, there were 12 cases accompanied with laryngeal stenosis and one case accompanied with tracheal stenosis. In terms of etiological factor, all patients were cataloged into two groups, i.e. 25 cases with chemo-causis and 2 cases with trauma. The repair methods included colon interposition in 20 patients, free jejunum transplantation in 6 patients and pectoralis major muculocutaneous flap in 1 patient. In total 12 cases of laryngeal stenosis, sternohyoid myofascial flap was applied in 8 cases, and sternohyoid myofascial flap and epiglottis were applied in 4 cases. RESULTS: Swallow function recovered in 25 cases and failed in 2 cases. Laryngeal function recovered totally in 10 patients and partially in 2 patients with laryngeal stenosis. The patient with tracheal stenosis recovered and decannulated. CONCLUSION: According to the foci of pharyngo-esophageal stenosis, the colon interposition, the free jejunum transplantation and the pectoralis major myocutaneous flap can be applied respectively to restore normal physiological function.

[Surgical management of primary cervical tracheal cancer].

OBJECTIVE: To review the experience of surgical treatment of primary cervical tracheal cancer. METHODS: Six patients with primary cervical tracheal cancer were treated surgically from January 1997 to April 1999. The trachea anastomosis, platysmamyocutaneous flap combiend with the facial flap of the sternohyoid muscle, sternocleidomastoid myoperiosteal flap and the pectoralis major muculocutaneous flap were applied to restore the defects of cervical trachea. By pathology, there were two squamous cell carcinomas, three adenoid cystic carcinomas, and one adenocarcinoma. RESULTS: Six cases were decannulated from 23 days to 3 months after operation. The length of follow-up was more than 3 years. Five cases have stable airway by fiberscope and good voice after decannulation and there is no recurrence. One case died of lung metastasis 2 years after operation. CONCLUSION: Trachea anastomosis is suited for small partial defect. The platysmamyocutaneous flap combined with the facial flap of the sternohyoid muscle, sternocleidomastoid myoperiosteal flap and the pectoralis major muculocutaneous flap are ideal transplant for cervical tracheal reconstruction.