Investigation of skin prick test on 2707 patients with allergic rhinitis in Wuhan area / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the distribution of common allergens in patients with allergic rhinitis (AR) from 2006 to 2010 in Wuhan area, and provide the objective evidence for the prevention and treatment of AR.</p><p><b>METHODS</b>The medical records of skin prick test (SPT) performed on 2707 AR patients from 2006 to 2010 were retrospectively analysed, and the positive rate of different allergens and changing trends in this time were compared. SPSS 17.0 software was used to analyse the data.</p><p><b>RESULTS</b>There were significant differences among the Dermatophagoides pteronyssinus positive rate (χ(2) = 12.11, P < 0.05) and Dermatophagoides farinae positive rate (χ(2) = 11.11, P < 0.05) in the past 5 years. Meanwhile, there was an upward trend in the positive rate of dust mite, which the positive rate of Dermatophagoides pteronyssinus increased from 84.5% in 2006 to 90.5% in 2010 (χ(2) = 6.88, P < 0.05), positive rate of Dermatophagoides farinae increased from 81.5% in 2006 to 89.0% in 2010 (χ(2) = 9.68, P < 0.05); There were significant differences among the Mugwort and Ragweed positive rate of 5 years (χ(2) = 194.10, P < 0.05; χ(2) = 67.06, P < 0.05). There were significant differences among the mold I and mold II positive rate of 5 years between (χ(2) = 18.95, P < 0.05; χ(2) = 36.62, P < 0.05). Meanwhile, there was an upward trend in the positive rate of mold and fluctuant trend in the positive rate of spring-pollen.</p><p><b>CONCLUSIONS</b>Nearly five years, dust mites is still the most common allergens in AR patients, presenting upward trend; the positive rate of mold presenting upward trend; the positive rate of wormwood and guinea wood presenting downward trend; the positive rate of pollen presenting fluctuant trend.</p>

Activation of nuclear factor-kappaB and aberrant gene expression of interleukin 6 in middle ear cholesteatoma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To establish the activation of NF-kappaB in middle ear cholesteatoma, discuss the relationship of NF-kappaB and the gene expression of IL-6 and explore the pathogenesis of middle ear cholesteatoma.</p><p><b>METHODS</b>Ten cases of cholesteatoma and 6 cases of normal external meatal skin were obtained from middle ear surgery. The NF-kappaB DNA binding activity and the mRNA level of IL-6 in these two kinds of tissues were detected by electrophoretic motility shift assay (EMSA) and Reverse transcription polymerase chain reaction (RT-PCR) respectively. The relationship of NF-kappaB DNA binding activity and the mRNA level of IL-6 were analyzed.</p><p><b>RESULTS</b>The NF-kappaB DNA binding activities of cholesteatoma [(15.9 +/- 8.2)%] were higher than those in normal skin [(1.36 +/- 0.94)%, t = 3.502, P < 0.05]. The expression of IL-6 mRNA was increased significantly in patients with cholesteatoma, as compared with that in the control specimens (t = 2.166, P < 0.05) and had a significant positive correlation with NF-kappaB DNA binding activity (r = 0.752, P < 0.05).</p><p><b>CONCLUSIONS</b>The IL-6 mRNA expression in cholesteatoma is closely related with the activity of NF-kappaB. It is tempting to speculate that NF-kappaB play a key role in the activation of cytokine in cholesteatoma.</p>

Short hairpin RNA silencing of vascular endothelial growth factor, telomerase reverse transcriptase, and Bcl-xl expression in human laryngeal squamous carcinoma xenografted in nude mice / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the biological impact of pEGFP-shVEGF-shTERT-shBcl-xl expression in human laryngeal squamous carcinomas xenografted in nude mice and the related antitumor mechanism.</p><p><b>METHODS</b>A recombinant plasmid vector containing 3 different short hairpin RNA (shRNA) segments including pEGFP-shVEGF-shTERT-shBcl-xl was constructed and directly injected into the grafted tumors of human laryngeal squamous carcinoma in nude mice. The mRNA and protein expressions were determined by real-time RT-PCR and Western blot respectively. Apoptosis was determined by TUNEL assay using a commercial kit. Intratumoral microvessel density (MVD) was assessed by immunhistochemistry.</p><p><b>RESULTS</b>On the 14th days after the final treatment, mRNA and protein expression of VEGF, TERT, and Bcl-xl were markedly suppressed. The tumor sizes were significantly smaller than those in the other two group, with an overall tumor inhibition ratio of 91.2%. MVD counts in the pEGFP-shVEGF-shTERT-shBcl-xl treated group were significantly lower than those of the other two groups, along with increased apoptotic cells.</p><p><b>CONCLUSIONS</b>The data showed that inhibition of VEGF, TERT, Bcl-xl expression by RNAi technique induces cellular apoptosis and suppresses the growth of laryngeal squamous carcinoma in vivo. VEGF, TERT and Bcl-xl may be involved in the development of laryngeal cancers. The findings suggest a synergistic tumor therapeutic effect through simultaneous inhibition of the three genes. Multi-target RNA interference may provide a powerful strategy against human laryngeal cancers.</p>

Expression and activation of nuclear factor-kappaB in middle ear cholesteatoma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the expression and activation of NF-kappaB in middle ear cholesteatoma.</p><p><b>METHODS</b>The protein expression of NF-kappaB p65 in 21 middle ear cholesteatoma tissues and 8 normal external ear canal skin obtained in middle ear surgery were examined by immunohistochemistry; NF-kappaB DNA binding activity in these two kinds of tissues were also detected by electrophoretic motility shift assay (EMSA). The influence of cholesteatoma debris on the NF-kappaB DNA binding activity of HaCat cell were further analyzed.</p><p><b>RESULTS</b>All epithelial cell of cholesteatoma revealed a relatively abundant plasma expression of NF-kappaB p65 protein, among which 12 cases showed nuclear positive expression. In contrast,the normal skin epithelium only revealed a sparse plasma distribution of NF-kappaB protein. The levels of NF-kappaB p65 protein expression in the epithelium of middle ear cholesteatoma tissue and normal skin were 0.168 +/- 0.051, 0.088 +/- 0.019 (t = 4.211, P < 0.01), respectively. The NF-kappaB DNA binding activities of cholesteatoma [(16.5 +/- 10.1)%] were also higher than those in normal skin [(1.38 +/- 1.24)%, t = 3. 600, P = 0.014]. The NF-kappaB DNA binding activity of HaCat cell increased when exposed to cholesteatoma debris in a dose dependent manner.</p><p><b>CONCLUSION</b>NF-kappaB might be an important factor which was involved in the occurrence and development of cholesteatoma.</p>

Differentially expressed gene profiles of cochlea in rats induced by acute and chronic sodium salicylate injection / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the mechanism of electrophysiologic changes caused by different type of sodium salicylate injection.</p><p><b>METHODS</b>Decapitated three group rats ( acute injected, chronic injected and normal rats ) separately, dissected the temporal bones to collect cochlea, and the otic capsules were removed. Then the cochlear materials from each groups were pooled and homogenized respectively, extracted the total RNA, obtained cDNA from purified total RNA by reversed transcription, cDNA were transcripted to cRNA probes in vitro. Hybridized the cRNA probes with tester chip to evaluate the quality of probes, if good, hybridized the probes with real chip. Obtained three gene expression profiles of different groups of cochlea Analyzed the differentially expressed genes among three groups by SOM. Analogized the SOM result to electrophysiologic changes. Then analyzed the genes in clusters of analog results by Gene Ontology. Then the genes in clusters of analog results were analyzed by Gene Ontology. Hsp27 was chosen to validate the result of gene chip using real time quantitative reverse transcription PCR ( RTQ RT-PCR).</p><p><b>RESULTS</b>The probes was good, and the chip hybridization results was credible. We obtained 6 clusters genes by SOM analysis, in which we choose cluster 3 and cluster 4 as candidate cluster. There were 46 genes in cluster 3 and 30 genes in cluster 4 employing GO analysis, which involved in cell communication, cell motility, metabolism, immune response and nerve ensheathment, et al. The result of RTQ RT-PCR showed high concordance with that of gene chip.</p><p><b>CONCLUSION</b>It's a new method to study the mechanism of electrophysiologic changes caused by sodium salicylate by gene chip and SOM analysis.</p>

Inhibition of growth and proliferation of Hep-2 cells by targeting human telomerase reverse transcriptase mRNA using RNA interference technology / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of RNA interference by targeting human telomerase reverse transcriptase (hTERT) mRNA in the larynx cancer cell line, Hep-2.</p><p><b>METHODS</b>The primary structures of hTERT cDNA were found in GenBank. Then the structure analysis were done according to RNAi strategy which determined the specific base sequences to design shRNA plasmid. Two types of plasmid, pshRNA1 and pshRNA2, involved in fluorescein gene were synthesized based on the specific base sequences. Control pshRNA3, a random sequence, and control pshRNA4, without additional specific sequence were also constructed. Cells were treated daily with pshRNA1-4 or normal culture medium respectively. The pshRNA1-3 was identified by electrophoresis. After administration of pshRNA1-4, fluorescence expression was detected by confocal microscopy, the expression of hTERT of the transfected cells was determined by Western blotting, telomerase activity was measured by TRAP-PCR ELISA, cell viability was determined by MTT assay, morphological changes and apoptosis were examined by inverted microscope and TUNEL respectively.</p><p><b>RESULTS</b>There was a 400 bp balteum in pshRNA1-3 after cut by SalI, which was identical with the size of the objective gene. Many cells presented green fluorescence after being treated by pshRNA1-4, but there are much more dead green fluorescent cells in the pshRNA1 and pshRNA2 group. hTERT protein and telomerase activity was significantly decreased after treated by pshRNA1 or pshRNA2. It was observed that treatment with pshRNA1 or pshRNA2 in the presence of a valid transfection reagent could reduce cell viability of Hep-2 cells within 96 h (P < 0.01). Under the same culture conditions, cells grew more sparsely and the number of apoptotic cell increased significantly.</p><p><b>CONCLUSIONS</b>shRNA plasmid directed against human telomerase reverse transcriptase can effectively transfect Hep-2 cells. shRNA targeted hTERT gene can significantly inhibit the growth and proliferation of Hep-2 cells, which results in apoptotic cell death. RNA interference may be a promising strategy for the treatment of laryngeal cancer.</p>