Effects of bleomycin A5 on caspase-3, P53, bcl-2 expression and telomerase activity in vascular endothelial cells.

OBJECTIVE: The aim of this study was to investigate the molecular mechanism of bleomycin A5 in inducing the apoptosis of human umbilical vein endothelial cells (ECV304). MATERIALS AND METHODS: ECV304 cells were cultured and passaged, and then were divided into control group and three treatment groups. The later three groups were treated with 15, 75, and 150 µg/ml bleomycin A5 for 24 hours, respectively. The expressions of caspase-3, p53, and bcl-2 in ECV304 cells were detected by flow cytometry, and the activity of telomerase was determined using telomere repeat amplification protocol (TRAP)-silver staining method. RESULTS: After treatment with different concentrations of bleomycin A5, the expression of caspase-3 in ECV304 cells was increased. It was significantly decreased with the increase of bleomycin A5 concentration, but the difference between 75 µg/ml and 150 µg/ml groups was not significant. Bleomycin A5 could significantly increase the expression of p53, with concentration dependence. It had no obvious effect on bcl-2 expression. There was high expression of telomerase in control group. After treatment with different concentration of bleomycin A5, the telomerase activity was significantly decreased. CONCLUSION: Bleomycin A5 can increase caspase-3 and p53 levels and inhibit telomerase activity to induce ECV304 apoptosis.

Role of AQP1 in inner ear in motion sickness.

Inner ear is critical for the development of motion sickness (MS). The present work was designed to test the role of aquaporins (AQPs) in inner ear in MS. After repetitive stimulus of rotation, the MS symptom was steadily alleviated in mice. After repetitive stimulus of rotation, several AQPs mRNA levels including AQP1, AQP2, AQP3, AQP4, AQP6, AQP7, and AQP9 in the inner ears of mice were analyzed. It was found that AQP1 mRNA level was increased remarkably, which was reconfirmed by Western blotting analysis. In addition, the relationship between AQP1 expression and MS sensitivity was studied and it was shown that AQP1 mRNA level was negatively related to MS index in mice. We sought to examine the function of AQP1 in inner ear using an RNAi approach to reduce the AQP1 protein expression in vivo. It was first observed that AQP1 knockdown in inner ear resulted in a significant increase of MS sensitivity in mice. In conclusion, after repetitive stimulus of rotation, the alleviation of MS symptom in mice was, at least in part, due to the upregulation of AQP1 expression in inner ear. In addition, the sensitivity to MS in mice was, at least in part, dependent on the expression of AQP1 in inner ear. AQP1 in inner ear plays an important role in the development of MS, and might be a potential target for the prevention or management of MS.

[Glottic measurement and vocal evaluation after three surgical techniques in the treatment of bilateral vocal cord paralysis].

OBJECTIVE: To evaluate postoperative glottic area and vocal quality of three various surgical techniques for treating bilateral vocal cord paralysis, including laser arytenoidectomy (Group A, 24 cases), reinnervation of the posterior cricoarytenoid muscle by phrenic nerve (Group B, 9 cases) and arytenoidectomy accompanying lateral cordopexy by extralaryngeal approach (Woodman's procedure, Group C, 13 cases). METHODS: 46 cases suffered from bilateral recurrent laryngeal nerve injury were included in our study. The pre-postoperative glottic measurement and vocal acoustic parameters were analyzed. RESULTS: The decannulated cases in group A and group B and group C were 22, 8, 13 respectively. The post-operative mean maximal glottic area was (47.2 +/- 7.4) mm2, (78.3 +/- 16.0) mm2, (48.1 +/- 6.5) mm2 respectively. Group B cases glottic area was larger than that of group A and group C (t value were 4.46 and 3.85, P value were 0.000 and 0.001). No significant difference was found between group A and group C (t = 1.68, P = 0.101). After surgery, in group A, 17 cases voice quality was the same compared with that of before surgery, and 7 cases voice quality had become worse; In group B, the voice quality had become better in 5 cases, completely recovered in 1 case, and had not change in 3 cases; In group C, the voice quality had become deteriorated in 10 cases and no change in 3 cases. And in group B, ipsilateral diaphragm paralysis in 9 cases after surgery, whose vital capacity and forced vital capacity had decreased to 72%-84%, 76%-84% of that before the surgery respectively; and the diaphragm mobility had recovered by 35%-76% respectively, while vital capacity and forced vital capacity had become 93%-97%, 91%-98% of that before the surgery. In Group B, all cases' pulmonary function was normal half a year postoperatively. CONCLUSIONS: Reinnervation of the posterior cricoarytenoid muscle by phrenic nerve seems to be best procedure with better post-operative voice and larger glottic area. Although the sufficient airway for decannulation can be acquired in Group A and Group C, but most of patients in Group A had pre-operative vocal level and badly abnormal in Group C.

[Intratumor bleomycin-A5 injection under electrolaryngoscope for treatment of large laryngopharyngeal and laryngeal hemangioma].

OBJECTIVE: To study the therapeutic effect of bleomycin-A5 injection under eletrolaryngoscope for treatment of large laryngopharyngeal and laryngeal hemangioma. METHODS: Intratumor bleomycin-A5 injection under eletrolaryngoscope was performed in 18 cases of large laryngopharyngeal and laryngeal hemangioma with surface anaesthesia, for totally 7 to 14 (mean 10.2) injections in each case. RESULTS: Twelve patients were cured and 6 showed obvious improvement. Follow-up of the patients for over one year found no recurrence of the hemangioma. CONCLUSION: Bleomycin-A5 injection is a minimal invasive procedure for treatment of large laryngopharyngeal and laryngeal hemangioma, causing less pain and better preserving the laryngeal function without the necessity of tracheotomy.

[Glottic measurement and vocal evaluation before and after adult arytenoidectomy].

OBJECTIVE: To study minimal glottic area which can acquire sufficient airway for decannulation and maximal glottic area which can maintain preoperative vocal function on adult bilateral vocal cord paralysis with unilateral arytenoidectomy. METHODS: Sixteen adult received microscopic right arytenoidectomy with Diomed-25 laser under general anaesthesia and sustained laryngoscope from September 1998 to February 2003. The pre-postoperative glottic measurement and vocal acoustic parameters were analyzed. RESULTS: The postoperative maximal glottic area, maximal posterior glottic width and maximal opening angle between bilateral vocal cords of the 15 decannulated cases were (45.93 +/- 6.56) mm2, (4.97 +/- 0.73) mm and (24.34 +/- 4.74) degrees respectively. Compared with preoperative period, there were significant difference. Pre-postoperative acoustic parameters (Jitter, Shimmer, harmonics-noise ratio) were analyzed and no significant difference were found (P > 0.05, but there are significant difference in NNE (normalized noise energy) and MPT (maximum phonation time) (P < 0.05). Except for three cases whose postoperative glottic area were more than 50. 1 mm2, there were no significant difference in pre and postoperative NNE (P > 0.05). The minimal glottic area decannulated was 38.0 mm2. The more opening maximal glottic area, the greater of NNE because of bigger closing gap. Noticeable increase of NNE was observed when maximal opening glottic area was up to 50.1 mm2. CONCLUSIONS: The minimal glottic area for decannulation should be 38.0 mm2, and the maximal glottic area for maintaining preoperation vocal function should be 50.1 mm2 on adult bilateral voca cord paralysis with laser aryntenoidectomy.

[Comprehensive prognostic value of spontaneous and evoked electromyography in laryngeal paralysis].

OBJECTIVE: To determine the comprehensive prognostic value of spontaneous and evoked electromyography (EMG) in laryngeal paralysis. METHODS: The characteristics of laryngeal EMG of 91 cases with unilateral vocal cord paralysis (VCP) after thyroid surgery were assessed. All cases were divided into four groups according to the interval of laryngeal EMG after onset, which were group one (2 months shorter, n = 13), group two (2 to 4 months, n = 23), group three (4 to 6 months, n = 36), group four (6 months longer, n = 19). The waveform morphology and the amplitude of laryngeal EMG and the highest evoked compound muscular active potential (CMAP) of thyroarytenoid muscles were examined and analyzed during voluntary tasks. The potential amplitude was showed by the percentage of that of the healthy lateral. The criterion of evaluation on evoked potential was attained by calculating statistical confidence interval. RESULTS: The highest evoked CMAP in group one was significantly lower than that of the others (P < 0.05), but no significant difference was observed between group two and group three (P > 0.05), so group two and group three were analyzed together. There were 2 recovered cases and 11 unrecovered cases in group one. On the basis of this criterion that a positive prognosis for laryngeal recovery was indicated when the evoked CMAP presented and there was no misdirect generated potential, correct prognostic rate was 92% (12/13). There were 11 recovered cases and 48 unrecovered cases in group two and three. The highest evoked CMAP was much higher in the recovered than in the unrecovered, significant difference was observed between them (P < 0.001). On the basis of the criterion that a positive prognosis for laryngeal recovery was indicated when the highest evoked CMAP was higher than 26. 4%, correct prognostic rate was 90% (53/59). On the basis of the criterion that a positive prognosis for laryngeal recovery was indicated when there was no misdirect generated potential and the highest evoked CMAP was higher than 26.4%, correct prognostic rate was 93% (55/59). When the interval from onset to laryngeal EMG recovering was longer than 6 months, none of these patients had return of vocal cord mobility whatever were the outcomes of laryngeal EMG. CONCLUSIONS: Correct prognostic rate can be improved if the prognosis of VCP in different courses is judged respectively by analyzing comprehensively spontaneous and evoked EMG.

[Diagnosis and management of sulcus vocalis].

OBJECTIVE: To clarify the pathophysiology of sulcus vocalis and to develop a more rational approach to treatment. METHODS: Twenty-nine cases of sulcus vocalis patients were divided into three classification: Type I is a physiologic variant and no dysphonia (11 cases). Types II (sulcus vergeture, 13 cases) and III (sulcus vocalis, 5 cases) are characterized by severe dysphonia and loss of vibratory activity. Eighteen cases of dysphonia were treated by surgery and phonation training. The operations included fat injection into vocal cords (9 cases of types II and 1 cases of types III, including 1 case of types III of second operation), fat implantation into sulcus vergeture after incision (4 cases of types II and 1 cases of types III) and undermining of the mucosa and sulcus vocalis resection (4 cases of types III, including 1 case of second operation). Phonatory function and video laryngostroboscopic data were evaluated before and after surgery and phonation training treatment in 18 patients. The mean follow-up time was 15.3 months. RESULTS: Ten cases of types II had excellent results after fat injection into vocal cords (n = 6) and fat implantation into sulcus vergeture after incision (n = 4). Three cases of types II improved after fat injection into vocal cords. Three cases of type III had excellent results after sulcus vocalis resection. One case of type III had excellent results by Second operation (sulcus vocalis resection) after fat injection into vocal cord. One case of type III improved by Second operation (fat injection into vocal cords) after fat implantation into sulcus vergeture after incision. No postoperative complications were noted. CONCLUSION: Accurate classification of sulcus vocalis is important and then adapt treatment to different types. Fat implantation into sulcus vergeture to type II and sulcus vocalis resection to type III were the best choice methods.

[Nerve exploration and decompression for traumatic recurrent laryngeal nerve injuries induced by thyroid gland surgery].

OBJECTIVE: To show the findings of recurrent laryngeal nerve injury exploration and find out therapeutic effects, indications and timing of nerve decompression for traumatic recurrent laryngeal nerve injury induced by thyroid gland surgery. METHODS: In this study there were 87 patients with recurrent laryngeal nerve injury, including 65 for nerve exploration and 22 for nonsurgical treatment. During nerve exploration, the types, severity of laryngeal nerve injuries and laryngeal muscular mass were studied. Nerve decompression was performed in these 14 patients whose compressing sutures or compression due to cicatricial hypertrophy were received nerve decompression. RESULTS: Injuries caused by thyroid gland operations mostly are of suture ligation (43%) and nerve severance (48%); simple scar compression was found only in 6 cases (9%). Atrophy of the laryngeal muscles was not very serious in patients with a course less than 6 months. In 10 patients with a course less than three months, nerve decompression restored normal functional abductor and abductor motion of the vocal cord in 9 patients and had no effects in one. Although functional motion of vocal cord was not seen in one case with a course less than 3 months and 4 cases between 3 and 5 months, the mass and tension of the reinnervated vocal cord became much the same as the contralateral normal vocal cord, thus resuming symmetric vibration of the vocal cords and physiological phonation. Although nonsurgical treatment improved severe hoarseness, it didn't restore normal functional motion of the vocal cord and normal voice. CONCLUSIONS: Nerve exploration showed a primary rule for recurrent laryngeal nerve injury induced by thyroid gland surgery. Early and mid-stage recurrent laryngeal nerve exploration and decompression may restore normal motion of the glottis, and it suggested laryngeal delayed reinnervation may help patients with a course more than 6 months.