In vitro and in vivo effect of 5-FC combined gene therapy with TNF-α and CD suicide gene on human laryngeal carcinoma cell line Hep-2.

This study was aimed to investigate the effect of combined cancer gene therapy with exogenous tumor necrosis factor-alpha (TNF-α) and cytosine deaminase (CD) suicide gene on laryngeal carcinoma cell line Hep-2 in vitro and in vivo. Transfection of the recombinant eukaryotic vectors of pcDNA3.1 (+) containing TNF-α and/or CD into Hep-2 cells resulted in expression of TNF-α and/or CD gene in vitro. The significant increase in apoptotic Hep-2 cells and decrease of Hep-2 cell proliferation were observed using 5-FC treatment combined with TNF-a expression by CD/5-FC suicide system. Moreover, bystander effect was also observed in the TNF-α and CD gene co-expression group. Laryngeal squamous cell carcinoma (LSCC) mice model was established by using BALB/c mice which different transfected Hep-2 cells with pcDNA3.1 (+) containing TNF-α and/or CD were applied subcutaneously. So these mice are divided into four groups, namely, (1)Hep-2/TIC group; (2)Hep-2/CD group; (3)Hep-2/TNF-α group; (4)Hep-2/0 group. At day 29 after cell inoculation, volume of grafted tumor had significant difference between each two of them (P<0.05). These results showed that the products of combined CD and TNF-α genes inhibited the growth of transplanted LSCC in mice model. So by our observed parameters and many others results, we hypothesized that 5-FC combined gene therapy with TNF-αand CD suicide gene should be an effective treatment on Laryngeal carcinoma.

Middle frontal horizontal partial laryngectomy (MFHPL): a treatment for stage T1b squamous cell carcinoma of the glottic larynx involving anterior vocal commissure.

OBJECTIVE: The therapeutic effect of middle frontal horizontal partial laryngectomy (MFHPL) in treating stage T1b squamous cell carcinoma of the glottic larynx involving anterior vocal commissure (AVC) was compared with that of the anterior frontolateral vertical partial laryngectomy (AFVPL). The feasibility and practical significance of MFHPL in clinical application was discussed in the present study. METHODS: From January 1996 to January 2010, a total of 65 patients diagnosed with stage T1bN0M0 glottic laryngeal cancer were treated with MFHPL or AFVPL. The postoperative complications, glottic reconstruction, recurrence rate, voice quality and survival rates were evaluated and compared between two treatments. RESULTS: AFVPL and MFHPL were performed in 34 and 31 patients, respectively. Flexible fiberoptic laryngoscopy revealed that in the MFHPL-treated patients the reconstructed glottis was spacious and symmetric. In contrast, AFVPL treatment resulted in irregular glottic area with poor symmetry and tubular glottis. The incidence of postoperative laryngeal stenosis significantly differed between the MFHPL- and AFVPL-treated groups (P = 0.025). No significant difference was detected in the 3- and 5-year overall- or tumor-free survival rates between two treatments. The Voice Handicap Index (VHI) and maximum phonation time (MPT) after surgery were 51.0±12.99 and 12.42±3.44 sec in the AFVPL-treated group; while in the MFHPL-treated patients they were 31.81±7.48 and 7.65±1.98 sec, respectively. Both differences in VHI (P = 0.012) and MPT (P = 0.024) were significant between two treatments. CONCLUSIONS: MFHPL was comparable to AFVPL with respect to postoperative complications, recurrence rate and survival rates, but possessed advantages over AFVPL in terms of the incidence of laryngeal stenosis and voice quality. Our study indicated that MFHPL has a potential value in clinical practice of treating stage T1b squamous cell carcinoma of the glottic larynx involving AVC.

Supracricoid partial laryngectomy with cricothyroidopexy: a treatment for anterior vocal commissure laryngeal squamous carcinoma.

BACKGROUND: The clinical efficiency and functional outcomes of supracricoid partial laryngectomy (SCPL) with cricothyroidopexy (CTP) were compared with those of the traditional SCPL with cricohyoidoepiglottopexy (CHEP) in treating laryngeal squamous carcinoma involving anterior vocal commissure (AVC). METHODS: From January 2000 to June 2009, 50 patients diagnosed with early- or intermediate-stage (T1b-T3 classification) glottic cancer involving AVC were treated with SCPL-CHEP or SCPL-CTP. Postoperative complications, local recurrence, survival rate, and speech performance were compared between these 2 surgical procedures. RESULTS: Patients undergoing SCPL-CHEP or SCPL-CTP manifested similar levels of postoperative complications, tumor recurrence, and survival rates. However, the SCPL-CTP group showed significantly lower Voice Handicap Index (VHI) scores, higher maximum phonation time, and improved glottic reconstruction and closure than the SCPL-CHEP group. CONCLUSION: The SCPL-CTP procedure better preserves postoperative speech performance than the SCPL-CHEP procedure, underscoring the moderate effectiveness of SCPL-CTP as a treatment for laryngeal squamous carcinoma involving AVC.

Anesthesia for tracheobronchial foreign bodies removal via self-retaining laryngoscopy and Hopkins telescopy in children.

This study attempted to explore suitable anesthetic methods used for removal of tracheobronchial foreign body (FB) via self-retaining laryngoscopy and Hopkins telescopy in children. 92 cases had undergone FB removal via self-retaining laryngoscopy and Hopkins telescopy or rigid bronchoscopy in our hospital since 2006, of which 56 cases were under intravenous anesthesia and endotracheal intubation with muscle relaxation (IAEI with MR), and the other 36 cases were under intravenous anesthesia with spontaneous breathing (IASB). Operative parameters and intraoperative vital signs were analyzed. Tracheobronchial foreign body was successfully removed in 87 cases, and not found in the other 5 cases. SpO(2) was below 90% transiently in 41 cases, 29 cases of which were under IAEI with MR and 12 cases were under IASB. Laryngospasm and choke were found in 12 cases under IASB. Vital signs including P(ET)CO(2) and heart rate were stable in all the cases. The mean surgical time, anaesthetic induction and recovery time of IAEI with MR via self-retaining laryngoscopy group were (5.69 ± 3.43) min, (9.68 ± 1.66) min and (26.13 ± 6.94) min, IASB via self-retaining laryngoscopy group were (21.35 ± 17.25) min, (13.71 ± 3.79) min and (24.64 ± 5.44) min, IAEI with MR via rigid bronchoscopy group were (10.20 ± 5.01) min, (10.31 ± 3.56) min and (25.13 ± 6.21) min, and IASB via rigid bronchoscopy group were (25.35 ± 13.25) min, (14.71 ± 3.61) min and (26.22 ± 5.65) min. It's a new and wonderful surgical procedure that combining self-retaining laryngoscopy and Hopkins telescopy for removal of tracheobronchial foreign body. IAEI with MR is suitable for bronchial FBA cases via them, while IASB is better for tracheal FBA or complicated cases.

Removal of tracheobronchial foreign bodies via suspension laryngoscope and Hopkins telescope in infants.

OBJECTIVES: Tracheobronchial foreign body aspiration is a life-threatening accident in infants, and is still a formidable clinical emergency to both otorhinolaryngologists and anesthesiologists. In this study, we attempted to assess the safety and ease of tracheobronchial foreign body removal in infants via suspension laryngoscopy and Hopkins telescopy under general anesthesia with endotracheal intubation. METHODS: The retrospective clinical study from 2006 to 2010 included 50 infants with foreign body aspiration, of whom 35 underwent suspension laryngoscopy and Hopkins telescopy and the other 15 underwent rigid bronchoscopy. All of the procedures were under general anesthesia with endotracheal intubation. RESULTS: All of the patients underwent temporary extubation. The foreign body was successfully removed in 46 cases and was not found in the other 4 cases. The mean operation time in the rigid bronchoscopy group was 13.20+/-9.01 minutes, and that in the Hopkins telescopy group was 5.79+/-3.54 minutes. The oxygen saturation level was below 90% in 17 cases, of which 7 were in the rigid bronchoscopy group and 10 were in the Hopkins telescopy group. The vital signs, including the partial pressure of carbon dioxide in expiratory gas and the heart rate, were stable in all cases. CONCLUSIONS: Foreign body removal in infants via suspension laryngoscopy and Hopkins telescopy under general anesthesia with endotracheal intubation should be promoted, since it is relatively safe and easy for both anesthesiologists and otorhinolaryngologists to perform and has a remarkable success rate.

[DJ-1 expression in laryngeal squamous cell carcinoma and its relationship with tumor recurrence and metastasis].

OBJECTIVE: To detect the expression of DJ-1 in laryngeal squamous cell carcinoma (LSCC) and to study the relationship between DJ-1 expression and clinical indexes of LSCC. METHODS: The expressions of DJ-1 protein in 71 LSCC samples and 9 cases control samples from laryngeal mucosa tissues of non-LSCC patients were detected using streptavidin peroxidase immunohistochemistry staining and the relationships between DJ-1 protein expression and clinicopathologic characteristics were analyzed. RESULTS: (1) The positive expression rate of DJ-1 protein in LSCC was 85.9%(61/71), which was significantly higher than the rate (55.5%, 5/9) in control laryngeal mucosa tissues (P < 0.05). (2) DJ-1 expression was related to tumor recurrence (P < 0.05), but not to sex, age, primary cancer position, T stage, clinical stage, lymph node metastasis and tumor differentiation. Tumor recurrence rate (53.3%) in the patients with higher expression of DJ-1 protein was higher than the rate (26.8%) in the patients with lower expression of DJ-1 protein (χ(2) = 5.164, P < 0.05). (3) With Kaplan-Meier curves and Cox regression analysis, the cumulative 5-year survival rates were correlated with DJ-1 expression levels in laryngeal cancer tissues or cervical lymph node metastasis (all P < 0.05), but not to sex, age, primary cancer position, T stage, clinical stage and tumor differentiation. CONCLUSIONS: The expression of DJ-1 protein in LSCC is higher than that in control laryngeal mucous tissues. Overexpression of DJ-1 is associated with poor overall survival in LSCC patients.

[Rapid eye movement-related and none rapid eye movement-related classification in obstructive sleep apnea hypopnea syndrome].

OBJECTIVE: To study the value of a new measurement that divided obstructive sleep apnea-hypopnea syndrome (OSAHS) into rapid-eye-movement (REM) related and non-rapid-eye-movement (NREM) related subgroups. METHODS: According to Siddiqui classification, 137 adult patients with OSHAS were diagnosed as REM-related OSAHS [REM apnea hypopnea index (AHI)/NREM AHI > 1] or NREM-related OSAHS (REM AHI/NREM AHI < 1). Polysomnographic data were compared and discussed. RESULTS: (1) There were 72 cases defined as REM-related OSAHS (52.6%) and 65 cases defined as NREM-related OSAHS (47.4%). (2) In all cases, total AHI and NREM AHI in REM-related OSAHS were significantly lower than those in NREM-related OSAHS, while lowest arterial oxygen saturation (LSaO2), REM LSaO2 and NREM LSaO2 were significantly higher than those in NREM-related OSAHS (t were -6.466, -7.638, 3.426, 2.472, 4.873 respectively, P < 0.05). No significance was found in sleep structure, REM AHI and REM LSaO2 between REM-related and NREM-related OSAHS (P > 0.05). (3) Given the severity of OSHAS, the constituent ratio of REM-related OSAHS decreased (77.8%, 61.5%, 37.3%) from mild to severe OSAHS, while that of NREM-related OSAHS rose (22.7%, 38.5%, 62.7%; chi² = 16.996, P < 0.01). In mild and moderate groups, REM LSaO2 of REM-related OSAHS was significantly lower than those in NREM-related OSAHS (t were -4.273 and -2.136, P < 0.05), while the differences of total AHI and LSaO2, NREM LSaO2 between these two types were not significant. In severe group, AHI in NREM-related OSAHS was significantly higher than that in REM-related OSAHS, while LSaO2, REM LSaO2 and NREM LSaO2 was significantly lower than those in REM-related OASHS (t were -4.943, 2.574, 1.996, 3.571, P ≤ 0.05). (4) There was no significance in sleeping latency and efficiency between REM-related and NREM-related OSHAS. CONCLUSIONS: REM-related OSHAS mainly exists in mild and moderate OSHAS, while NREM-related one mainly exists in severe OSHAS. NREM-related OSAHS may be more severe in AHI and hypoxia than REM-related one. Whenever obstructive apnea happened in REM or NREM period, its impacts on sleep structure, efficiency and latency have no difference.

Evaluation of long-term toxicity of Ad/hIFN-, an Adenoviral vector encoding the human interferon-gamma gene, in nonhuman primates.

Interferon-gamma(IFN-gamma) plays an important role in the immunomodulation and growth inhibition of many tumor cells, but its clinical application is limited by its systemic toxicity. Ad/hIFN-gamma, a nonreplicating adenoviral vector encoding human IFN-gamma, has been reported to inhibit tumor growth in vitro and in a xenograft model. In this study, the long-term toxicity of Ad/hIFN-gamma was assessed in cynomolgus macaques (Macaca fascicularis). Thirty animals were enrolled into 5 groups, and administered intramuscularly, respectively, Ad/hIFN-gamma (3.3 x 10(10), 3.3 x 10(11), or 3.3 x 10(12) VP/kg), Ad/LacZ (vector control, 3.3 x 10(11) VP/kg), or excipient 3 times per week for 8 weeks, followed by a 4-week recovery period. At 12 weeks all experimental animals appeared generally healthy, and there were no statistically significant differences in body weight, urinalysis, hemogram, blood biochemistry, and electrocardiogram results between the treatment and control groups. No significant toxic effects were noted on macroscopic and microscopic examinations of organs and tissues. Preliminary investigation of the immunotoxicity of Ad/IFN-gamma indicated that anti-adenoviral and anti-hIFN-gamma antibodies were generated. These data demonstrate that long-term, high-dose intramuscular administration of Ad/IFN-gamma was not notably toxic and might be safe for clinical therapeutic use.

[Analysis of relevant factors causing laryngeal stenosis after partial laryngectomy].

OBJECTIVE: To investigate the clinical relevant factors causing laryngeal stenosis after partial laryngectomy. METHODS: A retrospective study was carried out to review the history clinical data from 138 patients of partial laryngectomy in the First Affiliated Hospital of Sun Yat-Sen University between January 1994 to October 2004. The clinical relevant factors causing laryngeal stenosis were included as follows: age, sex, TNM stage, tumor site, extension of thyroid cartilage defect, extension of larynx parenchyma defect, reconstruction method, laryngeal dilator, duration of using antibiotics, postoperative radiotherapy, lung infection, gastroesophageal reflux, diabetes. Multivariate stepwise logistic regression model was used for the analysis. RESULTS: Of 138 cases after partial laryngectomy, stenosis developed in 25 cases. The occurrence rate was 18.1%. In multivariate analysis, it was confirmed that the following factors correlated to laryngeal stenosis, i. e, extension of thyroid cartilage defect (chi2 = 4.323, P = 0.038), postoperative radiotherapy (chi2 = 6.002, P = 0.014), lung infection (chi2 = 4.220, P = 0.040), and gastroesophageal reflux (chi2 = 5.614, P = 0.018). CONCLUSIONS: The clinical relevant factors causing laryngeal stenosis after partial laryngectomy were multiple. Statistical analysis showed that extension of thyroid cartilage defect, postoperative radiotherapy, lung infection and gastroesophageal reflux were the risk factors which may cause laryngeal stenosis.

[Meta analysis of relationship between laryngeal tumor of chinese and human papillomavirus infection].

BACKGROUND & OBJECTIVE: Researches showed that the infection of human papillomavirus (HPV) was closely related to laryngeal tumor, but those results existed great differences because of different research methods. In this study Fisher's and Meta analysis were used to synthetically evaluate relationship between laryngeal benign and malignant tumors of Chinese and infection of different types of HPV. METHODS: A total of 123 references about relationship between laryngeal benign and malignant tumors of Chinese and infection of different types of HPV were collected from CBMdisc (Chinese Biomedical Literature Analysis and Retrieval System for Compact Disc). There were 11 references accorded with research criteria which were case-control study, and had detected HPV16 or HPV6/11 by polymerase chain reaction (PCR). Fisher and Meta analysis were used to quantitatively and qualitatively analyze these references synthetically. RESULTS: Mean detection rates of HPV16 in normal laryngeal tissue, laryngeal carcinoma (LC), laryngeal papilloma(LPa), laryngeal polyp (LPo) from all references were 10.8%,35.2%,27.5%,5.0%, respectively; and those of HPV6/11 were 8.7%, 18.6%, 61.6%, 21.9%, respectively. The infection of HPV16 in LC were significantly higher than those in LPa, and LPo (P< 0.005); the combined odds ratio (ORc) for HPV16 infection in LC was 2.8 (1.7-4.7) times that in LPa, and 12.7 (4.2-38.8) times that in LPo; mean positive rate of HPV16 in LC was 22.0% (12.2%-31.8%) higher than that in LPa, and 39.0% (19.4%-58.6%) higher than that in Lpo. The infection of HPV6/11 in LPa was significantly higher than that in LC (P< 0.005); OR(c) for HPV16 infection in LPa was 16.4 (5.6-48.1) times that LC; mean positive rate of HPV16 in LPa was 56.0% (34.4%-75.6%) higher than that in LC. There was no significant difference of HPV6/11 infection between LC and LPo (P >0.05). CONCLUSION: The infection of HPV16 may enhance risk of development of laryngeal carcinoma; and the infection of HPV6/11 may enhance risk of development of laryngeal papilloma.

[Investigation and analysis of quality of life for patients after laryngectomy].

OBJECTIVE: To institute and test the quality of life for laryngectomy, and evaluate the feasibility of above questionnaire in patients after laryngectomy. METHODS: A questionnaire of quality of life for laryngectomy was instituted. An investigation was conducted in patients treated by partial (36, Group A) or total (45, Group B) laryngectomy for laryngeal cancer with above questionnaire. The feasibility, reliability and validity of the questionnaire was evaluated, the quality of life and the influencing factors in laryngectomy were also analyzed. RESULTS: The questionnaire of quality of life for laryngectomy consists of 22 items, covering six aspects. The questionnaire has a comparatively good reliability, and the criterion-related validity and content validity of the questionnaire is valid. The correlation coefficient of split-half method and Cronback's alpha of questionnaire were 0.842 and 0.889 respectively. The composite quality of life scores of group A were significant higher than those of group B ( P < 0.001). The differences in physical function, laryngeal function, psychological state, the ability of living independently were statistically significant (P < 0.001; P < 0.001; P < 0.01; P < 0.05) as well. Seven factors including operative modality, tumor stage, postoperative complication, coexisted disease, family incomes, voicing modality and wearing tracheo-cannula were related to postlaryngectomy quality of life. CONCLUSIONS: The questionnaire of quality of life for laryngectomy has speciality in laryngectomy and a comparatively good reliability and validity, and it is suitable for quality of life research in patients after laryngectomy.

[Inhibition of hydroxycamptothecin on laryngeal squamous carcinoma cell line].

BACKGROUND & OBJECTIVE: Hydroxycamptothecin(HCPT) has wide antitumor spectrum and high inhibition rate of tumor cell line and xenografts. Recently,no studies on the treatment of laryngeal squamous carcinoma with HCPT were reported. Hence,this study were designed to investigate the inhibition of O-HCPT [ring-opened form(O-HCPT)] and C-HCPT[ring-closed form(C-HCPT)] on laryngeal squamous carcinoma cell line(Hep-2 cell line)and its mechanism. METHODS: The cytotoxicity of O-HCPT and C-HCPT on Hep-2 cells was measured by MTT assay and cell cycle was detected using flow cytometry (FCM). The growth state of Hep-2 cell xenografts treated with 10 mg/kg HCPT (O-HCPT or C-HCPT) was observed. The doubling time and the tumor inhibition rate were calculated. RESULTS: The growth inhibition of O-HCPT and C-HCPT on Hep-2 cell depended on concentration. The IC(50) were 0.69 and 0.48 micromol/L, respectively. After treated with high concentration of HCPT, the cell cycle was arrested in S phase and then apoptosis were obviously induced. At the low concentration of HCPT, the cell cycle was slightly arrested in G(2)+M phase. Compared with control group, the xenografts of O-HCPT (10 mg/kg) treated group grew slowly and tumor doubling time prolonged. There was significant difference in the tumor volumes of two HCPT-treated group (P< 0.001) and the tumor inhibition rate was 77.0%. All mice in C-HCPT (10 mg/kg) treated group died of toxicity. CONCLUSION: The result showed that O-HCPT and C-HCPT had obvious cytotoxicity to laryngeal squamous carcinoma cells which mechanism was HCPT arrest cell cycle in S phase and induce cell apoptosis. O-HCPT has slight toxicity effect and can be used as chemotherapeutic agent for laryngeal squamous carcinoma, but C-HCPT had strong toxicity.