Role of folate receptor 1 in paclitaxel-resistance of nasopharyngeal carcinoma cells / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To provide experimental evidence for the folate receptor 1 (FOLR1) mediated targeted cancer therapy and resistance reversal, the FOLR1 expression differences in nasopharyngeal carcinoma cells (CNE-1) and immortalized normal nasopharyngeal cells (NP69) and the correlation between FOLR1 expression and paclitaxel resistance index in nasopharyngeal carcinoma were investigated.</p><p><b>METHODS</b>The expressions of FOLR1 in CNE-1, CNE-1/Taxol (paclitaxel-resistance cells) and NP69 was detected by cDNA microarray, reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot and immunocytochemistry. Proliferation inhibition rates of CNE-1 and CNE-1/Taxol cells were measured by colony formation assay after treated by short interfering RNA (siRNA) of FOLR1.</p><p><b>RESULTS</b>The expressions of FOLR1 gene in CNE-1/Taxol cells and CNE-1 cells were 2636.0 and 176.0, respectively. The expression of FOLR1 was not detected in the NP69 by semi-quantative RT-PCR and Western blot. The high expression of FOLR1 in CNE-1/Taxol was verified by semi-quantative RT-PCR, and its expression level was positively correlated to the degree of drug-resistance (r(2) = 0.8719). The results were also validated by Western blot and immunocytochemistry. The sensitivity of CNE-1/Taxol to paclitaxel significantly increased after inhibition of FOLR1 gene expression by siRNA, and its IC(50) value was decreased by 59.6% (t = 6.92, P < 0.01).</p><p><b>CONCLUSIONS</b>The expression of FOLR1 is closely related to the occurrence of NPC and Taxol resistance. FOLR1 gene may be one of the important target molecules in NPC treatment and reversion of the paclitaxel-resistance in NPC.</p>

Expression of human beta-defensin after endoscopic sinus surgery for chronic sinusitis / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the expressions of human beta-defensin 1 and 2 (hBD-1 and hBD-2) in nasal mucosa before and after the endoscopic sinus surgery and investigate the effects of hBD-1 and hBD-2 on the healing process after the surgery.</p><p><b>METHODS</b>The patients undergoing endoscopic sinus surgery for chronic sinusitis were divided into 3 groups according to the response to the surgery, namely cured group, response group and non-response group. With those from healthy control subjects as the control, nasal mucosa samples were collected from the patients at 1 week, 2 weeks, 1 month, 3 months and 6 months after the surgery for detection of hBD-1 and hBD-2 mRNA and protein expressions by RT-PCR and Western blotting.</p><p><b>RESULTS</b>hBD-1 and hBD-2 were expressed in both normal and chronic sinusitis mucosa, but the expression levels varied significantly between the individuals. The expression levels of hBD-2 was significantly correlated to the patients' response to the surgical treatment (P<0.05). hBD-1 showed slight differences between the individuals, but was not associated with the patients' prognosis.</p><p><b>CONCLUSION</b>The expressions of hBD-2 mRNA and protein are significantly increased in patients with good response to endoscopic sinus surgery for chronic sinusitis, suggesting the value of hBD-2 as an indicator of the patients' prognosis.</p>

Expression and significance of human beta-defensin in recurrent nasal polyps / 中南大学学报(医学版)

OBJECTIVE@#To observe the expression of human beta-defensin-1 (hBD-1) and human beta-defensin-2 (hBD-2) in recurrent nasal polyps, and to investigate the role of beta-defensin in the recurrent nasal polyps.@*METHODS@#Tissues of nasal polyps was obtained from 10 patients with nasal polyps undergoing endoscopic sinus surgery, recurrent nasal polyps from 10 patients 6 months after surgery, nasal mucosa from 10 recovered patients with nasal polyps postoperatively and,10 control subjects. hBD-1 mRNA and hBD-2 mRNA levels of tissue specimens in all groups were detected by reverse transcriptase-polymerase chain reaction (RT-PCR).@*RESULTS@#There was no significant difference in hBD-1 mRNA level between the 4 groups (P>0.05). Expression of hBD-2 mRNA was detected in patients with nasal polyps and recurrent nasal polyps, but rare in the recovered patients and the control subjects.@*CONCLUSION@#hBD-1 is a constitutive expression and hBD-2 is an induced expression. beta-Defensin may play an important role in forming the nasal polyps.

Protective effect of NT-3 gene mediated by hydroxyapatite nanoparticle on the cochlea of guinea pigs injured by excitotoxicity / 中南大学学报(医学版)

OBJECTIVE@#To transfect the recombinant plasmid enhancement type green fluorescent protein C2- neurotrophic factor-3 (pEGFPC2-NT3) into the spinal ganglion cells(SGCs) of guinea pigs' cochlea injured by the excitotoxicity of hydroxyapatite particle (HAT), to inject the recombinant plasmid pEGFPC2-NT3 into the guinea pigs' cochlea, and to observe the expression of pEGFPC2-NT3 and the protective effect of pEGFPC2-NT3 on SGCs of the cochlea in guinea pigs.@*METHODS@#The recombinant plasmid pEGFPC2-NT3 with gene-green fluorescent protein was established. Kanic acid (KA) was injected into guinea pigs'cochleae and the excitotoxicity model was established. After a week the recombinant plasmid was transferred into SGCs of guinea pigs'cochlea treated with HAT. The following week the expression of NT-3 was examined by the immunohistochemical method, and the morphology of SGNs was observed under the electronic microscope after 4 weeks, in the mean time the changes of auditory brain-stem response (ABR) were examined.@*RESULTS@#The excitotoxicity models were established successfully. NT-3 expression in the intracytoplasm of SGNs was observed by the immunohistochemical method 1 week after the injection, the morphologic damages of SGNs lessened under the electronic microscope after 4 weeks. ABR was partly restored, compared with ABR after the injury of the excitotoxicity.@*CONCLUSION@#On the 7th day, NT3 gene transferred by HAT through the scala tympani can lessen the excitotoxicity of SGCs after KA was injected into the guinea pigs cochlea.

Determination of differentially expressed proteins and it's significance among chronic sinusitis, nasal polyps and normal nasal mucosa / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the differentially expressed proteins among chronic sinusitis, nasal polyps and normal nasal mucosa by means of proteomic technology, and select the candidate biomarkers of chronic sinusitis and nasal polyps.</p><p><b>METHODS</b>Proteins extracted from chronic sinusitis, nasal polyps and normal nasal mucosa were separated and the differentially expressed proteins were identified by series of proteomic tools, including immobilized pH4-7 gradient two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis, modified coomassie brilliant blue staining, images scanning by the Image Scanner apparatus, PDQuest analysis software, peptide mass fingerprinting based on matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) by in-gel digestion extract, and Mascot searching in NCBInr and SWISS-PROT databases.</p><p><b>RESULTS</b>The 2-DE patterns with high resolution and reproducibility were obtained. The protein spots separated and visualized in chronic sinusitis, nasal polyps and normal nasal mucosa gel were 1020 +/- 40, 1112 +/- 10 and 1008 +/- 25, respectively. And the match rates were (93 +/- 2)%, (95 +/- 1)% [see text] (90 +/- 3)% respectively. Thirteen differentially expressed spots were found from chronic sinusitis, nasal polyps and normal nasal mucosa gel. We selected and recommend Keratin 8 and APOA1 proteins as candidate biomarkers of nasal polyps, and PLUNC protein, PACAP protein, NKEF-B and SOD as candidate biomarkers of chronic sinusitis.</p><p><b>CONCLUSIONS</b>The differentially expressed proteins among chronic sinusitis, nasal polyps and normal nasal mucosa can be efficiently and relatively reliably identified via the techniques of proteomics. These techniques will play a very important role in the researches for new objective indicators possibly employed in the future classifying, staging and prognosis.</p>

Causes and revision endoscopic sinus surgery for recurrent sinusitis / 中南大学学报(医学版)

OBJECTIVE@#To investigate the causes of revision endoscopic sinus surgery (RESS), and to evaluate the clinical effect and experience of RESS in the patients with recurrent sinusitis.@*METHODS@#Before the revision surgery, 168 patients (225 sides) with recurrent sinusitis were examed by CT scans and nasal endoscopy. All patients were operated by revision endoscopic sinus surgery.@*RESULTS@#Among the 168 patients, 95 cases (121 sides) had incomplete middle turbinate, 51 cases (75 sides) had maxillary sinus ostium stenosis, 85 cases (117 sides) had nasal cavity adhesion, 48 cases (57 sides) had uncinate process residual, 38 cases (65 sides) had ethmoid hyperostosis, and 25 cases had nasal septum deviation. One hundred and six patients (134 sides) were cured, 40 patients (56 sides) were improved, useful 22 patients (35 sides) were ineffective.@*CONCLUSION@#The most common surgical causes of failures in endoscopic sinus surgery are the disease area remnant, nasal cavity adhesion, maxillary sinus ostium stenosis, uncinate process residual, ethmoid hyperostosis, and nasal septum deviation. Revision endoscopic sinus surgery is a useful way to treat recurrent sinusitis. Preoperative CT scan, correct choice of the anatomic marks of orientation and direction function are the key to operation successful.

Establishment of 2-dimensional gel electrophoresis map and analysis of proteomics from human nasal polyps / 中南大学学报(医学版)

OBJECTIVE@#To establish 2-dimensional polyacrylamide gel electrophoresis (2-DE) map from human nasal polyps and normal nasal mucosa, and to identify differential expression proteins of 2-DE map.@*METHODS@#Samples of nasal polyps and nasal mucosa (each sample group containing 7 cases) were obtained. The total proteins were extracted and separated by immobilized pH gradient (IPG)-based 2-DE. The silver-stained 2-DE was scanned with digital Imagescanner and analyzed with ImageMaster 2-DE Elite 4.01 software. To obtain peptide mass fingerprint (PMF) of differential protein spots, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used. The PMF was searched in Swiss-Prot and TreMBL database by Pept-Ident software, to identify differential expression proteins.@*RESULTS@#The well-resolved, reproducible 2-DE maps of nasal polyps and nasal mucosa were established. For the polyps tissues, the average proteins spot of three 2-DE maps was 825+/-78; and 682+/-96 spot was matched with the average matching rate of 82.7%. The average deviations of matched spot position were (1.13+/-0.16) mm in IEF direction and (1.45+/-0.21) mm in SDS-PAGE direction, respectively. For the nasal mucosa tissues, the average proteins spot of three 2-DE maps was 936+/-62; and 821+/-78 spots were matched with the average matching rate of 87.7%. After comparing the 2-DE maps of nasal polyps and nasal mucosa tissues, the protein spots were 1,458 and 1,617 respectively; and 1,026 protein spots were matched. Forty differential expression protein spots were incised from silver staining gel randomly and digested in the gel by TPCK-Trypsin. Thirty-four PMFs were obtained by MALDI-TOF-MS and 24 differential proteins were identified.@*CONCLUSION@#The well-resolved, reproducible 2-DE maps of human nasal polyps and nasal mucosa have been successfully established. Certain differential proteins related to the pathogenesis of human nasal polyps are identified.