AIM: To quantify cyclooxygenase-2 (COX-2) mRNA in carcinoma of larynx and evaluate the correlation between the quantity of COX-2 mRNA and clinical staging or histological grade. METHODS: The expression of COX-2 mRNA in 30 cases of carcinoma of larynx tissue and adjacent non-cancerous tissues were evaluated by PCR, which includes a fluorescence dye , SYBR green Ⅰ, and the sequence specific primer. The GAPDH was used as control. RESULTS: The specificity of products was proved to be COX-2 and GAPDH by the analysis of the melting curve of the amplified products and agarose gel electrophoresis. The expression of COX-2 mRNA was detected in all cancerous tissues of 30 patients (100%), but only in 12 adjacent non-cancerous tissues of 30 patients (40%). The N_ COX value of carcinoma of larynx tissue and adjacent non-cancerous tissues was 16.54?13.27 and 9.24?6.91, respectively, and the expression levels of COX-2 mRNA elevated significantly in laryngeal squamous cell carcinoma tissue and there were significant correlation between the expression levels of COX-2 mRNA and clinical stage or histological grade. CONCLUSION: The expression of COX-2 mRNA in carcinoma of larynx can be determined by real-time PCR technique. An increase in COX-2 mRNA may be associated with carcinogenesis of carcinoma of larynx, and it may be useful as a biomarker in laryngeal cancer.

The inhibitory effects of two kinds of selective cyclooxygenase-2 inhibitors on the proliferation of human carcinoma of larynx Hep-2 in vitro and their corresponding mechanisms were investigated. Hep-2 cells were cultured with two kinds of selective cyclooxygenase-2 inhibitors (Sc-58125 and Celecoxib) at various concentrations for 24 h. Morphological changes were observed under the phase microscopy and the growth suppression was detected by using MTT colorimetric assay. Apoptotic DNA fragments were observed by agarose gel electrophoresis, and the cell cycle and apoptotic rate were detected by flow cytometry (FCM) respectively. Hep-2 cells became rounded and detached from the culture dish after being treated with Celecoxib for 24 h, however, they remained morphologically unchanged with Sc-58125. Sc-58125 could increase G2 phase cells, whereas, Celecoxib rose G1 phase cells. Both of the two effects were dose-dependent. Moreover, the Hep-2 cells cultured with 50 micromol/L and 100 micromol/L Celecoxib showed obvious apoptosis, with the nuclear DNA of cells exhibiting characteristic DNA ladder. So Sc-58125 could inhibit the proliferation of Hep-2 cells by altering the G2 phase cells. However, Celecoxib had the same effect by changing the G1 phase cells and inducing apoptosis at higher concentration.
Antineoplastic Agents/pharmacology ; Apoptosis/drug effects ; Cell Proliferation/drug effects ; Cyclooxygenase 2 Inhibitors/*pharmacology ; Dose-Response Relationship, Drug ; Laryngeal Neoplasms/*pathology ; Pyrazoles/*pharmacology ; Sulfonamides/*pharmacology ; Tumor Cells, Cultured

AIM: To quantify cyclooxygenase - 2 (COX - 2) mRNA in carcinoma of larynx and evaluate the correlation between the quantity of COX - 2 mRNA and clinical staging or histological grade. METHODS: The expression of COX - 2mRNA in 30 cases of carcinoma of larynx tissue and adjacent non - cancerous tissues were evaluated by PCR, which includes a fluorescence dye , SYBR green Ⅰ , and the sequence specific primer. The GAPDH was used as control. RESULTS: The specificity of products was proved to be COX - 2 and GAPDH by the analysis of the melting curve of the amplified products and agarose gel electrophoresis. The expression of COX - 2 mRNA was detected in all cancerous tissues of 30 patients (100%), but only in 12 adjacent non - cancerous tissues of 30 patients (40%). The NCOX value of carcinoma of larynx tissue and adjacent non - cancerous tissues was 16.54 ± 13.27 and 9.24 ± 6.91, respectively, and the expression levels of COX- 2 mRNA elevated significantly in laryngeal squamous cell carcinoma tissue and there were significant correlation between the expression levels of COX - 2mRNA and clinical stage or histological grade. CONCLUSION: The expression of COX - 2 mRNA in carcinoma of larynx can be determined by real - time PCR technique. An increase in COX - 2 mRNA may be associated with carcinogenesis of carcinoma of larynx, and it may be useful as a biomarker in laryngeal cancer.

Cholesteatoma, Middle Ear ; Humans ; Male ; Young Adult

A strain of high-efficiency bioflocculant-producting bacteria,which was named TJ-3,was screened from sewage sludge.The strain was gram-negative and rod-shaped in physiological biochemical test and was identified as Pseudomonas aeruginosa by 16S rDNA Sequencing.According to the growth curve of the TJ-3 strain,the growth stabilization period is long so that Microbial Flocculants(MBF) production is stable.The MBF produced by the TJ-3 strain is able to flocculante kaolin suspension with 98.2%.The MBF activity distribution tests show that most of the active components of the bioflocculant exist in deposition after centrifufation.When pH is 8.5,1%(quality fraction) CaCl2 Solution dosing quantity is 3.5 mL,bioflocculant dosing quantity is 1.5 mL in 100 mL kaolin suspension,the bioflocculant has an optimal flocculation.

A survey on compliance with topical medication and the relevant factors were conducted in 190 patients with glaucoma in Miaohang Township, Baoshan County of Shanghai. All patients used 1 or more topical ocular hypotensive medications for at least 6 months and a special questionnaire was designed for the survey. The survey revealed that the overall non-compliance rate was 54. 7% (104/190) in this group of patients; which was closely correlated with age, medication application time and the extent of visual defect ( OR = 2. 550, 0. 225 and 0. 342, P ＜ 0. 05 ). Education levels, gender of patients, glaucoma type, number of daily medication, medication types and the stability of diseases were not correlated with the compliance.

[Summary] Microcephalic or Majewski's osteodysplastic primordial dwarfism type Ⅱ ( MOPD Ⅱ) is an extremely rare genetic disease mainly caused by pericentrin ( PCNT) gene mutations. This paper reported one 13-year-old boy, who was admitted because of the slow growth for more than 13 years and deepened skin color over six months. He was diagnosed as MOPD Ⅱ associated with a combination of growth hormone deficiency, type 2 diabetes, hypertension, acanthosis nigricans, multiple café-au-lait spots. On magnetic resonance imaging of brain, no vascular malformations such as aneurysms were shown. There were novel compound heterozygous mutations of PCNT gene in the patient, with the nonsense mutations of c. 502C > T ( p. Gln168 * heterozygous variation) and c. 3103C > T (p. Arg1035* heterozygous variation). His father carried a nonsense mutation c. 3103C > T ( p. Arg1035 *heterozygous variation ) and his mother had a nonsense mutation c. 502C > T ( p. Gln168 * heterozygous variation). After treatment with metformin for three months, his blood glucose returned to normal, and acanthosis nigricans was improved. It seems critical to evaluate the abnormal condition of blood vessels regularly for MOPD Ⅱpatients with PCNT gene mutations.

The inhibitory effects of two kinds of selective cyclooxygenase-2 inhibitors on the proliferation of human carcinoma of larynx Hep-2 in vitro and their corresponding mechanisms were investigated. Hep-2 cells were cultured with two kinds of selective cyclooxygenase-2 inhibitors (Sc58125 and Celecoxib) at various concentrations for 24 h. Morphological changes were observed under the phase microscopy and the growth suppression was detected by using MTT colorimetric assay. Apoptotic DNA fragments were observed by agarose gel electrophoresis, and the cell cycle andapoptotic rate were detected by flow cytometry (FCM) respectively. Hep-2 cells became roundedand detached from the culture dish after being treated with Celecoxib for 24 h, however, they remained morphologically unchanged with Sc-58125. Sc-58125 could increase G2 phase cells, whereas, Celecoxib rose G1 phase cells. Both of the two effects were dose-dependent. Moreover, the Hep-2 cells cultured with 50μmol/L and 100 μmol/L Celecoxib showed obvious apoptosis, with the nuclear DNA of cells exhibiting characteristic DNA ladder. So Sc-58125 could inhibit the proliferation of Hep-2 cells by altering the G2 phase cells. However, Celecoxib had the same effect by changing the G1 phase cells and inducing apoptosis at higher concentration.

To investigate the expression of vasoactive intestinal peptide (VIP) and substance P (SP) in the cochlea of spontaneously hypertensive rat (SHR), and to assess the function of VIP and SP in the cochlea following the damage of hypertension, hearing thresholds of ABR were observed and the fixative (4% paraformaldehyde) was pumped through the circulatory system. Adult Wistar rats (3 months, n=20) served as the control group and SHRs (3 months, n=20) as the hypertension group. Bullas were taken out and cochleas were irrigated in vitro with the same fixative. The number of base turn's spiral ganglions in the sections was counted. The expression of VIP and SP were detected by SABC method and the images of the sections were analyzed. The number of base turn's spiral ganglsons in the hypertension group was significantly less than in the normal group (P<0.01). VIP and SP were expressed in the spiral ganglion cytoplasma and stria vascularis of the two groups. There were no significant difference in the expression of VIP and SP in spiral ganglion cytoplasma (P>0.05) between the two groups. However, in stria vascularis the expression of VIP in the hypertension group was higher than in the normal group (P<0.05), and no significant difference in SP was found between the two groups. It was suggested that VIP not only contributed to the regulation of the cochlea microcirculation, but also made the neurotransmitter in the pathway of the auditory system. However, SP made only the neurotransmitter in the pathway of the auditory system.
Animals ; Cochlea ; metabolism ; Hypertension ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Wistar ; Substance P ; biosynthesis ; genetics ; Vasoactive Intestinal Peptide ; metabolism

Objective To analyze the clinical feature, diagnosis and treatment of Alstrom syndrome. Method The clinical data of a case of Alstrom syndrome and the result of her ALMS1 sequencing by the two generation sequencing were retrospectively reviewed. Results The 12 year and 10 month old female suffered from dilated cardiomyopathy, obesity, optic nerve diseases, sensorineural hearing loss, high blood glucose and irregular menstruation since one month of birth. Laboratory examination showed she had high testosterone level, hyperglycemia, hyperlipidemia and fatty liver. High-throughput sequencing confirmed there was ALMS1 gene mutation which includes hybrid frameshift mutations of c.5418delC and p.Y1807Tfs*23, and heterozygous nonsense mutation of c.10549C>T and p.Q3517*, and c.5418delC was a new variation reported for the first time. Conclusion Alstrom syndrome is an autosomal recessive genetic disease, which is characterized by multiple organ dysfunction and metabolic syndrome, and can be diagnosed by gene detection.