Effect of microbubbles on transcranial doppler ultrasound-assisted intracranial recombinant tissue-type plasminogen activator thrombolysis.

OBJECTIVES: The aim of this study was to evaluate the effect of microbubbles on the efficacy of transcranial doppler (TCD) ultrasound-assisted thrombolytic therapy of recombinant tissue-type plasminogen activator (rt-PA). METHODS: Male New Zealand white rabbits (n = 36) were randomly divided into an rt-PA group (n = 18) and an rt-PA plus microbubble group (n = 18). After the cerebral infarction model was constructed with autologous blood clots, rt-PA and rt-PA plus microbubble intervention were performed, respectively. The hemodynamic changes and infarct size of the two groups were recorded. In addition, the ELISA method was used to detect the level of nitric oxide (NO), superoxide dismutase (SOD), and malondialdehyde (MDA) in the brain tissue of the two-group graph model and high-sensitivity C-reactive protein (hs-CRP) in the serum. RESULTS: In the rt-PA group, the recanalization rate was 38.9% and the average infarct size was 11.8%. In the rt-PA plus microbubble group, the recanalization rate was 66.7% and the average infarct size was 8.2%. In addition, the average values for NO, SOD, MDA, and hs-CRP were 16.48 ± 5.39 µmol/L, 730.2 ± 9.86 U/mg, 0.92 ± 0.43 nmol/mg, and 8.56 ± 1.64 mg/L in the rt-PA group, respectively, and the average values were 9.18 ± 3.37 µmol/L, 426.2 ± 6.39 U/mg, 0.73 ± 0.44 nmol/mg, and 5.23 ± 0.94 mg/L in the rt-PA plus microbubble group, respectively. CONCLUSIONS: The addition of microbubbles enhanced the effects of TCD-assisted rrt-PA thrombolysis.

Relationship between microemboli in the internal carotid artery and the occurrence of ischemic stroke after transient ischemic attack.

Microembolic signals (MES) have been reported to be an independent risk factor for stroke and transient ischemic attack (TIA). We examined the relationship between MES in the internal carotid artery and the occurrence of ischemic stroke in patients with TIA. A total of 67 patients who had a TIA were examined with transcranial Doppler ultrasonography to detect microemboli in the internal carotid artery 1, 3, and 7 days after admission, and 3 months after discharge. The relationship between the presence of MES and the subsequent occurrence of ischemic stroke was the primary outcome of interest. 35.8% (24/67) of patients were MES(+). During follow-up, ischemic stroke occurred significantly more frequently in patients who were MES(+) compared with patients who were MES(-) (6/24; 25.0% versus 2/43; 4.7%, p=0.021), as did TIA (11/24; 45.8% versus 4/43; 9.3%). MES(+) status was significantly associated with the occurrence of ischemic stroke after adjusting for age, sex, hypertension, diabetes mellitus, and drug therapy (odds ratio: 8.30; 95% confidence interval: 1.37-50.42; p=0.021). The positive and negative predictive values of MES status for predicting ischemic stroke were 25.0% and 95.4%, respectively. The presence of microemboli in the internal carotid artery appears to be an important risk factor for the occurrence of ischemic stroke after TIA. The MES(+) rate in patients with transient ischemic attack with severe internal carotid artery stenosis is markedly higher than in patients without internal carotid artery stenosis.

Effects of microbubbles on transcranial Doppler ultrasound-assisted intracranial urokinase thrombolysis.

INTRODUCTION: To evaluate the efficacy of microbubbles in transcranial Doppler ultrasound (TCD)-assisted urokinase thrombolysis. MATERIALS AND METHODS: Male New Zealand white rabbits (N=32) were randomly divided into 2 groups, a urokinase group and a combined urokinase plus microbubble group. The middle cerebral artery (MCA) was occluded by injecting autologous blood clots through the carotid artery. In the urokinase plus microbubble group, sulfur hexafluoride (SonoVue) microbubbles were injected intravenously immediately after intravenous injection of urokinase. The 2 groups were monitored by TCD from before until 2h after thrombolysis, and the hemodynamic changes and infarct size were recorded. RESULTS: The urokinase alone group had 1 case of complete recanalization and 4 cases of partial recanalization (recanalization rate, 31.3%). The urokinase plus microbubble group had 3 cases of complete recanalization and 6 cases of partial recanalization (recanalization rate, 56.3%). The average size of the infarction foci was 13.9% in the urokinase group and 9.1% in the urokinase plus microbubble group (P=0.025). Pathological examination revealed no cerebral hemorrhage in either group. CONCLUSIONS: The addition of microbubbles enhanced the effects of transcranial Doppler ultrasound-assisted urokinase thrombolysis.

[Construction and confirmation of a recombinant adenovirus vector of survivin].

OBJECTIVE: To construct a recombinant adenovirus of survivin vector and provid valuable reference for gene therapy of laryngeal cancer. METHODS: The survivin gene was cloned by PCR. After confirmation by enzyme restriction analysis and sequencing, the gene and the adenovirus vector were recombined together to construct the recombinant adenovirus vector. The recombinant adenovirus vector was confirmed via both sequencing and digestion restriction analysis, and then linearized and transfected into the HEK 293 cell line to generate recombinant adenovirus. RESULTS: The sequence analysis demonstrated that the survivin gene sequence was the same as published in the literature, suggesting that a recombinant adenovirus vector has been successfully constructed. CONCLUSIONS: A survivin recombinant adenovirus has been successfully constructed.

[Expression of survivin gene in thyroid carcinoma and its significance].

OBJECTIVE: To study the expression of survivin gene in thyroid carcinoma and its significance. METHODS: Samples of thyroid carcinoma were collected from 98 patients during operation. Western blotting and RT-PCR were used to detect mRNA and protein expression of survivin in the carcinoma tissue and the tissue around the carcinoma. TUNEL method was used to detect the apoptotic index (AI). RESULTS: Western blotting and RT-PCR methods showed similar results. The positive expression rates of survivin protein and mRNA in the thyroid carcinoma were both 68.37%; however, survivin was not expressed in all of the tissues around the thyroid carcinoma at both the mRNA and protein levels. The expression rate of survivin was closely related to the pathological typing, lymph node metastasis, and hematogenous metastasis. Apoptotic cells were found in all thyroid carcinoma. The mean AI of the surviving positive group was 0.48 +/- 0.38, significantly lower than that of the survivin negative group (2.98 +/- 0.12, P = 0.0082). CONCLUSION: Survivin gene is expressed in most of the thyroid carcinoma. The positive expression rate of survivin is significantly linked to cell proliferating activity, lymph node metastasis, and hematogenous metastasis. Survivin gene will be a new target of gene therapy in thyroid carcinoma.

[Supraglottic horizontal partial laryngectomies in 163 cases].

OBJECTIVE: To evaluate the clinical value of supraglottic horizontal partial laryngectomy. METHODS: One hundred and sixty-three patients with supraglottic laryngeal carcinoma were treated surgically by supraglottic horizontal partial laryngectomy from 1978 to 1998. There were 64 males and 99 females. Five cases were staged I, 95 staged II, 48 staged III and 15 staged IV. The surgical techniques were improved: The hyoid was removed conventionally; The outer perichondrium of thyroid cartilage was turned into the laryngeal cavity and sutured with the mucosa of laryngeal ventricle and the base of tongue was sutured to the reserved thyroid cartilage. Survival was evaluated using the Kaplan-Meier method. The differences between stages were tested by Los-Rank method. RESULTS: The 5-year survival rate were 100.0%, 77.9%, 54.2% and 33.3% for patients staged I to IV respectively, and were statistically significant (P = 0.0006) between different clinical stages. The 5-year survival rate were 73.1% and 45.5% in patients with cN0 and cN + respectively, and were statistically significant (P = 0.0132). The speech and swallowing functions were good after operation. The decannulation rate was 91.4%. The main causes of death were cervical lymph node metastasis in 40% (20/50) and laryngeal recurrence in 18% (9/50). The occult metastasis rate was 23.1% (30/130) and cervical metastasis rates of patients with cN1-3 75.8% (25/33), with a total metastasis rate was 33.7 (55/163). CONCLUSIONS: Supraglottic horizontal partial laryngectomy is very effective in eradicating disease and in preserving laryngeal function on condition that the indications were selected correctly. The selective lateral neck dissection was recommended for supraglottic carcinoma.

[Effect of survivin antisense oligodeoxynucleotide on carcinoma of larynx in vivo and in vitro].

OBJECTIVE: To observe the effect of survivin antisense oligodeoxynucleotide (ASODN) on the apoptosis of human carcinoma of larynx cell line Hep2 and the inhibitory rate in nude mice model so as to discuss the selective blocking activity of antisense technique on gene expression seeking a new way for gene therapy of carcinoma of larynx. METHODS: Antisense oligodeoxynucleotides survivin were transformed into human carcinoma of larynx cell line Hep2 by liposome Lipofectamine 2000. Within 72 h after transfection, 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay was used to detect cellular proliferation. Forty eight hours after transfection, reverse transcription-polymerase chain reaction (RT-PCR) assay was used to observe the expression of survivin gene, Western Blot assay for the protein, and terminal deoxynucleotide mediated nick end labeling (TUNEL) and flow cytometer for cellular apoptosis. RESULTS: Cellular inhibition rate of 72 h went up to 52. 5% and 71.4% at 1.0 micromol/L and 2.0 micromol/L value in Lipo-ASODN groups which differed statistically remarkably (P = 0.046), higher than that in controls in MTT assay (P =0. 003 and 0. 0004). Forty eight hours after transfection survivin gene expression in Lipo-ASODN groups were less than that in control group in RT-PCR assay. Survivin protein expression decreased in Western blot. In TUNEL assay, nuclear positive staining was observed and the apoptosis peak was observed in flow cytometer test, which were absent in controls. In nude mice of carcinoma of larynx model, the inhibitory rate in Lipo-ASODN groups got up to 48.1% and 61.3% higher than that of controls (P < 0.004 and 0. 0006), which differed remarkably (P = 0.032) in a dose-dependently way. CONCLUSIONS: The findings showed that the expression of survivin gene and protein induced cellular apoptosis in Hep2 cells after transfection of Lipo-ASODN and that the carcinoma of larynx in the nude mice model were inhibited by Lipo-ASODN which suggested that antisense technique can be an effective means in the gene therapy of carcinoma of larynx.

[Cervical metastasis in patients with T2-4 cN0 laryngeal carcinoma].

OBJECTIVE: To study the characteristics of the cervical lymph node metastasis in clinical N0 (cN0) patients with laryngeal carcinoma and its implication in clinical treatment. METHODS: 76 patients with laryngeal carcinomas of T2-4cN0 category were divided into two groups in random: 21(22 sides) radical neck dissection(RND) and 55(60 sides) functional neck dissection(FND) were performed. Lymph nodes were studied histologically according to the levels. RESULTS: On an average, 29.6 lymph nodes were obtained in one side of neck in RND group, and 24.7 in FND group(F = 3.145, P = 0.068). The occult metastasis rates were 33.3% (7/21) in RND group and 34.5% (19/55) in FND group. 25 of 26 patients (96.2%) who had positive nodes involved only the levels II and III. 2130 lymph nodes were obtained in all samples, 59 of 60 positive nodes(98.3%) were located in the level II and III. The 5 and 10-year survival rates of the two groups were 71.4% (15/21), 76.4% (42/55) and 61.9% (13/21), 68.9% (31/45), respectively with no statistical difference(chi 2 = 0.2394, P > 0.5; chi 2 = 0.3143, P > 0.05). Ipsilateral cervical recurrence rates in two groups were 9.5% (2/21) and 7.3% (4/55), respectively with no statistical difference (chi 2 = 0.1059, P > 0.900). 10-year mortalities with negative and positive cervical lymph nodes were 16.7% (7/42) and 62.5% (15/24) respectively, which had statistically difference (chi 2 = 14.4375, P < 0.005). CONCLUSION: The lateral neck (level II, III and IV) dissection may be suitable for the treatment laryngeal carcinoma patients with T2-4cN0.