[Show abstract][Hide abstract] ABSTRACT: The NLR pyrin domain containing 3 (NLRP3) inflammasome plays a crucial role in lung disease and may have a similar role in upper respiratory tract inflammation. We therefore constructed a C57BL/6 mouse model of acute rhinosinusitis induced by Staphylococcus aureus and investigated the role of the NLRP3 inflammasome in this model. Mice were classified as non-inoculated group (group A) and inoculated groups (groups B, C, D and E, sacrificed 1, 3, 7 and 14 days after inoculation, respectively). Hematoxylin-eosin staining showed that each group had inflammatory cell infiltration, except group A. The damage of the nasal mucosa was aggravated gradually over time. Western blot and immunofluorescence showed that the structural proteins of the NLRP3 inflammasome (NLRP3, ASC (apoptosis-associated speck-like protein containing CARD), procaspase-1) in groups B, C, D and E were increased gradually. But they were reduced in group B compared with group A, except for NLRP3. Western blot showed that the cleavage fragment of procaspase-1, p20 in groups B, C, D and E was increased gradually. Real-time PCR showed that the corresponding mRNAs of the structural proteins were changed the same as their proteins. IL-1β mRNA and mature IL-1β protein were increased gradually in groups A, B, C, D and E. These results indicate that NLRP3 inflammasome activation was associated with the acute rhinosinusitis, and that there was a positive correlation between the expression level of the NLRP3 inflammasome and the severity of acute rhinosinusitis.
Full-text · Article · Sep 2014 · International Journal of Molecular Sciences
[Show abstract][Hide abstract] ABSTRACT: Recent studies indicated that interleukin (IL)-17, growth-related oncogene (GRO)-α and IL-8 play an important role in the pathogenesis of nasal polyps. However, the effects of the increased amount of IL-17 and the production of GRO-α and IL-8 in human nasal polyp fibroblasts are not completely understood. This study aimed to determine the effects of the increased IL-17 on the changes of GRO-α and IL-8 expression in human nasal polyp fibroblasts and further investigate the mechanism of neutrophil infiltration in nasal polyps. Nasal polyp fibroblasts were isolated from six cases of human nasal polyps, and the cells were stimulated with five different concentrations of IL-17. Real-time fluorescence quantitative polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of GRO-α and IL-8. The mRNA of GRO-α and IL-8 was expressed in unstimulated controls and remarkably increased by stimulation with IL-17. Moreover, the levels of GRO-α and IL-8 produced by fibroblasts were increased gradually with the increases in IL-17 concentrations. The present study showed that nasal fibroblasts can produce GRO-α and IL-8, and their production is remarkably enhanced by IL-17 stimulation, thereby clarifying the mechanism of the IL-17 mediated neutrophil infiltration in nasal polyps. These findings might provide a rationale for using IL-17 inhibitors as a treatment for nasal inflammatory diseases such as nasal polyps.
No preview · Article · Aug 2014 · Journal of Huazhong University of Science and Technology
[Show abstract][Hide abstract] ABSTRACT: To study the effect of neonatal immunization with different dosage allergen on the immunity of mice when grown.
Fifty neonatal BALB/c mice were divided into 4 groups randomly and subcutaneous injected with different dosage of ovalbumin (OVA) on day 1, 8 and 15 after born [NS group(10): injected with saline alone; NS + AL group (10): injected with saline and AL(OH)3; small dosage (SD) group (15): injected with 10 microg OVA and AL(OH)3; large dosage (LD) group (15): injected with 1000 microg OVA and AL(OH)3]. The mice were then challenged using caudal vein injection on 5 weeks old (NS group and NS + AL group were challenged with saline, SD group and LD group were challenged with 100 microg OVA). The blood was collected 1 week later to examine OVA specific IgE, IgG1 and IgG2a. Mononuclear cells were drawn from the spleen and cultured. Concentration of IL-4, IFN-r, IL-10 was examined in the cultural supernatant. Flow cytometry was used to test the expression of CD4+ IL-4+, CD4+ IFN-gamma+, CD4 IL-10 cells.
It was found that concentration of OVA specific IgE (OVA-sIgE) in SD group (0.33 +/- 0.18) was significantly higher than that of NS (0.07 +/- 0.01) and NS + AL (0.09 +/- 0.04) group (t value was -3.46 and -3.21, all P < 0.01), and LD group (0.17 +/- 0.10) as well (t = 2.58, P < 0.05). The concentration of OVA-sIgE was higher in LD group than that of NS group (t = -2.53, P < 0.05), but similar with that of NS + AL group (t = -2.04, P > 0.05). Both the concentration of OVA-sIgG1 and sIgG2a was higher in SD and LD group than that of NS and NS + AL group (all P < 0.05). The concentration of IL-4, IFN-gamma and IL-10 in the cultural supernatant of spleen mononuclear was all higher in SD group than that of NS and NS + AL group (all P < 0.01). The ratio of IFN-gamma/IL-4 was significantly lower in SD group than that of NS and NS + AL group (t value was 2.14, 3.44, all P < 0.05), while the same ratio was higher in LD group than that of NS and NS + AL group (t value was -2.14, -1.61, all P < 0.05). Ratio of CD4+ IL-4+ cells was significant lower in LD group than that of SD group (P < 0.05), while it was not different with that of NS and NS + AL group (P > 0. 05).
Neonatal immunization with low dosage OVA could generate a specific immunity with Th2 direction, while with large dosage OVA could generate a specific immunity with Th1 direction.
No preview · Article · Jul 2013 · Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery
[Show abstract][Hide abstract] ABSTRACT: Computed tomography (CT) scan with three-dimensional (3D) reconstruction of the inner ear provides a more accurate image of the relationship of the electrode within the cochlear canal, with direct demonstration of electrode insertion depth in the cochlea in comparison with X-ray plain film.
This study was designed to evaluate the value of spiral CT scans with 3D reconstruction in determining the insertion site and depth of implanted cochlear implant electrodes.
A total of 172 cochlear implant recipients were involved in this study. The implanted electrodes of all patients were examined by X-ray plain film, and 157 cochlear recipients were examined by spiral CT scans with axial 1 mm image slices. The data from the CT scans were transferred to a workstation for 3D reconstruction (direct volume rendering) of the inner ear. The pseudocolor technique was used to display the electrode.
The insertion depth of the electrode could be evaluated indirectly by the X-ray plain film. In contrast, the stereoscopic images from a CT scan with 3D reconstruction of the inner ear demonstrated the shape, position, and insertion depth of the electrode more accurately.
No preview · Article · Nov 2011 · Acta oto-laryngologica
[Show abstract][Hide abstract] ABSTRACT: To evaluate the Integrated Cochlear Profile for Assessing Auditory Nerve-Auditory Pathway Integrity (ICP-API), as proposed by our group, in the selection of cochlear implant candidates.
The API of the candidates for cochlear implantation were assessed with the ICP-API, which consists of 5 categories including: audiological testing; radiological imaging study; ear-canal electric response audiometry; response to environmental sounds; speech development level. The auditory rehabilitation effects of the cochlear implantation receivers were evaluated postoperatively.
Sixty-six of 68 candidates who met the criteria of the ICP-API received cochlear implantation with improved hearing and speech development postoperatively. The remaining 2 of the 68 candidates were diagnosed with bilateral auditory nerves aplasia, and therefore failed cochlear implants were avoided.
The ICP-API is valuable and feasible for the selection of cochlear implant candidates. The API should be considered as one of the most important criteria for cochlear implant candidate selection.
[Show abstract][Hide abstract] ABSTRACT: A trans-superior meatus endoscopic approach to treat diseases in the sphenoid sinus and sellar area is a safe, minimally traumatic, and effective method.
To avoid complications, we explored the use of the superior meatus and superior turbinate in the endoscope approach to the sphenoid sinus and sellar area.
This was a retrospective analysis of the curative effect of the trans-superior meatus approach for diseases in the sphenoid sinus and sellar area in 138 cases.
All of 138 patients had successful operations and no serious complication occurred. All cases were followed up for a period of 1-3 years. No recurrence was found in 94 patients with isolated sphenoid sinus disease (sinusitis, mucoceles, or mycosis). Of 24 patients with pituitary adenoma, 17 patients had entire resection and no recurrence was found. Four patients had subtotal resection and three patients had partial resection with postoperative radiotherapy, and preoperative symptoms were improved. Of 13 cases of cerebrospinal rhinorrhea in the sphenoid sinus, 12 cases were successfully repaired by a single operation and 1 case was successfully repaired by a repeat operation. Among seven cases with decompression of the optic canal, four had obvious effect, two cases showed improvement, and there was no improvement in one case.
No preview · Article · Dec 2008 · Acta Oto-Laryngologica
[Show abstract][Hide abstract] ABSTRACT: PC cell-derived growth factor is a novel growth factor for tumor formation and progression. No comprehensive literature concerning PC cell-derived growth factor expression status and its biological function in squamous cell carcinoma, especially in the larynx, is, however, available. The target of this study is to evaluate the clinical significance of PC cell-derived growth factor and the potential of small interfering RNA-induced genetic silencing of PC cell-derived growth factor as a supplementary therapeutic way for laryngeal squamous cell carcinoma. A total of 146 primary laryngeal cancer, 108 adult laryngeal papilloma and 41 laryngeal leukoplakia samples, as well as 10 normal larynx tissues were investigated. The PC cell-derived growth factor mRNA level was examined by real-time polymerase chain reaction and protein localization by immunohistochemistry. The biological function of PC cell-derived growth factor was assessed by transfection of small interfering RNA PC cell-derived growth factor construction. The PC cell-derived growth factor protein levels and mRNA levels of the laryngeal squamous cell carcinomas were significantly higher than those of normal laryngeal tissues (P<0.001). Simultaneously, the difference in the levels of mRNA and protein between those of laryngeal precancerous lesions (papilloma/leukoplakia) and those of normal tissues was significant (P<0.05, P<0.05), whereas those of laryngeal precancerous lesions (papilloma/leukoplakia) were significantly lower than those of laryngeal squamous cell carcinomas (P<0.05, P<0.05). Strong PC cell-derived growth factor expression was associated with lymph node metastases in laryngeal squamous cell carcinoma (P<0.05). Functional studies on Hep-2 cell lines demonstrated that the attenuation of PC cell-derived growth factor expression levels led to diminished cell proliferation rates (P<0.001), anchorage-independent growth in vitro (P<0.001), tumor forming in vivo (P<0.01) and resistance to apoptosis (P<0.001). PC cell-derived growth factor is a pivotal autocrine growth factor in the tumorigenesis of laryngeal squamous cell carcinoma. Our findings also indicate that PC cell-derived growth factor is a logical and potential target for early diagnosis, specific therapy and prognosis of laryngeal squamous cell carcinoma.
No preview · Article · Jan 2007 · Anti-Cancer Drugs
[Show abstract][Hide abstract] ABSTRACT: Hair cells of inner ear are suggested to be inhibited by the activation of the alpha9-containing nicotinic acetylcholine (ACh) receptors (alpha9-containing nAChRs). Several studies have suggested that the native nicotinic-like ACh receptors (nAChRs) in hair cells display a significant permeability of Ca(2+) ions and unusual pharmacological properties. The activation of native nAChRs will initiate the hyperpolarization of hair cells by activation of the small conductance, Ca(2+)-activated K(+) channels (SK). In this work, the properties of the ACh-sensitive potassium current (IK(ACh)) in outer hair cells (OHCs) of guinea pigs were investigated by employing whole-cell patch-clamp. Followed by perfusion of ACh, OHCs displayed a rapid desensitized current with an N-shaped current-voltage curve (I-V) and a reversal potential of - 66 +/- 7 mV. The IK(ACh) was still present during perfusion of either iberiotoxin (IBTX, 200 nM) or TEA (5 mM) but was potently inhibited by apamin (1 muM), TEA (30 mM). The IK(ACh) demonstrated a strong sensitivity to alpha-bungarotoxin (alpha-BgTx), bicuculline and strychnine. These results suggested that OHCs display the well-known SK current, which might be gated by the alpha9-containing nAChRs. Two important changes were present after lowering the Ca(2+) concentration in the external conditions from 2 mM to 0.2 mM: one was a flattened N-shape I-V relationship with a maximum shifted toward hyperpolarized potentials from -20 approximately -30 mV approximately -40 to -50 mV, the other was a significant reduction in the agonist maximal response (percentage of maximal response 10.5 +/- 5.4). These results indicated that native nAChRs are both permeable to and modulated by extracellular Ca(2+) ions. Taken together, this work provides direct evidences that SK channels in OHCs of guinea pigs are gated by alpha9-containing nAChRs, which play an important role in the fast cholinergic efferent inhibition. This fast inhibition is both potently dependent on the permeability of Ca(2+) ions through the native nAChRs and modulated by Ca(2+) ions.
[Show abstract][Hide abstract] ABSTRACT: Death-associated protein kinase (DAPK) is a Ca/calmodulin-regulated serine/threonine kinase and a positive mediator of apoptosis. Loss of expression of the DAPK gene by aberrant promoter methylation may play an important role in cancer development and progression. The aim of this study was to investigate the frequency of gene promoter methylation of DAPK in nasopharyngeal carcinoma (NPC) and the effect of 5-Aza-2'-deoxycytidine (5-Aza-CdR), a demethylating agent, on CNE cells, a human nasopharyngeal carcinoma cell line, and on xenografts of CNE cells. Methylation-specific PCR and RT-PCR were used to determine the promoter methylation status and mRNA expression of the DAPK gene in NPC. Furthermore, CNE cells were treated in vitro and in vivo with 5-Aza-CdR to explore the effect of demethylating agents on DAPK mRNA expression and tumor growth. Hypermethylation of the DAPK gene promoter was found in 35 (76.1%) of 46 NPC samples. There was no significant difference in the promoter hypermethylation rate among samples from patients with different TNM stages. No promoter hypermethylation of the DAPK gene was found in all six chronic inflammatory nasopharyngeal tissue specimens. DAPK mRNA expression was not detected in NPC tumor specimens with promoter hypermethylation. However, DAPK mRNA expression was observed in unmethylated NPC tumors and in the chronic inflammatory nasopharyngeal tissue specimens. Promoter hypermethylation of the DAPK gene was found and no DAPK mRNA expression was detected in CNE cells. DAPK mRNA expression in CNE cells and xenografts could be restored by treatment with 5-Aza-CdR. The CNE cell xenografts of nude mice treated with 5-Aza-CdR were obviously smaller in tumor volume than those of nude mice treated with PBS. These results demonstrate that loss of DAPK expression could be associated with promoter region methylation in NPC. 5-Aza-CdR may slow the growth of CNE cells in vitro and in vivo by reactivating the DAPK gene silenced by de novo methylation.
No preview · Article · Apr 2006 · Anti-Cancer Drugs
[Show abstract][Hide abstract] ABSTRACT: Molecular biological studies have demonstrated that both muscarinic receptor subtypes and nicotinic receptor subunits were located in mammalian vestibular sensorineural epithelium. However, the functional roles are still unclear, with the exception of the well-known alpha9-containing nicotinic ACh receptor (alpha9nAChR). In this study, the properties of acetylcholine (ACh)-induced currents were investigated by whole-cell patch clamp technique in isolated type II vestibular hair cells (VHCs II) of guinea pig. VHCs II displayed a sustained, non-inactivating current when extracellular application of ACh. ACh-induced currents restored gradually and it took about 60 s to get a complete recovery. ACh-induced current was not affected by extracellular Na(+), but strongly affected by extracellular K(+) and Ca(2+). Depletion of the intracellular Ca(2+) stores by intracellular application of inositol 1,4,5-trisphosphate (IP3) or blocking of the release of intracellular Ca(2+) stores by intracellular application of heparin failed to inhibit this current. ACh-induced currents were inhibited by nifedipine, Cd(2+), tetraethylammonium (TEA), charybdotoxin (CTX), iberiotoxin (IBTX), atropine and d-tubocurarine (DTC), respectively, but not by apamin. In conclusion, ACh stimulates a large conductance, Ca(2+)-activated K(+) current (BK) in guinea pig VHCs II by activation of the influx of Ca(2+) ions, which is mediated by an ACh receptor that could not be defined to be the odd-number muscarinic receptor.
No preview · Article · Dec 2005 · Hearing Research
[Show abstract][Hide abstract] ABSTRACT: 5-Aza-2'-deoxycytidine (5-Aza-CdR) is an inhibitor of DNA methyltransferase; it may reactivate methylated antioncogene, therefore, inhibit the growth of cancer cells. This study was to observe the inhibitory effect of 5-Aza-CdR on the growth of human nasopharyngeal carcinoma (NPC) cells and xenografts in nude mice, explore the possible mechanisms, and search for new treatment target of NPC.
NPC cell line CNE cells were treated with 5-Aza-CdR; the methylation status of death-associated protein kinase (DAPK) gene was evaluated by methylation-specific polymerase chain reaction (PCR). The model of human NPC xenograft in nude mice was constructed and treated with 5-Aza-CdR; the xenograft growth in nude mice was observed, and the mRNA and protein expression of DAPK was detected by reverse transcription-PCR (RT-PCR) and immunohistochemistry.
No expression of DAPK mRNA was found in CNE cells and the xenografts in nude mice without treatment of 5-Aza-CdR. After treatment, the expression of DAPK mRNA in CNE cells and the xenografts was increased along with the increasing concentration of 5-Aza-CdR; the growth of CNE cells and the xenografts in nude mice were obviously inhibited, and the methylated DAPK gene was reactivated. Four weeks after treatment, no significant difference was found in body weight of nude mice between 5-Aza-CdR group and control group [(22.35+/-2.02) g vs. (21.68+/-2.14) g, t=0.011, P>0.05]; the volume of xenografts was significantly smaller in 5-Aza-CdR group than in control group [(195.32+/-27.57) mm(3) vs. (343.67+/-23.08) mm(3), t=10.11, P<0.01].
5-Aza-CdR may reactivate antioncogene silenced by de novo methylation, therefore, inhibit the growth of CNE cells in vivo and in vitro.
No preview · Article · Oct 2005 · Ai zheng = Aizheng = Chinese journal of cancer
[Show abstract][Hide abstract] ABSTRACT: Objective: To study the correlation between Ki67 and vascular endothelial growth factor(VEGF) and the significance of their expression
in laryngeal squamous cell carcinoma(LSCC). Methods: The expressions of Ki67 and VEGF in 40 cases of LSCC and 5 cases of normal laryngeal mucosa were examined by immunohistochemical
staining. Results: The expression levels of Ki67 and VEGF in LSCC tissue were higher than in normal laryngeal mucosa (Ki67: P<0.001, VEGF: P<0.001). The two indexes’ levels in patients of different age or different sex had no significant difference (P>0.05). They were higher in LSCC with metastasis of lymph nodes than in patients without metastasis (Ki67: P=0.034, VEGF: P=0.006). The expressions of the two genes elevated correspondingly along with the development of LSCC T stage (P<0.05). In addition, correlation analysis indicated that the expression of Ki67 had a positive correlation with VEGF in LSCC(r=0.823,
P<0.01). Conclusion: Ki67 and VEGF are objective indexes for the biological behavior of LSCC, and they might be helpful to the diagnosis, treatment
and prognosis of laryngocarcinoma.
No preview · Article · Jan 2005 · Chinese Journal of Cancer Research
[Show abstract][Hide abstract] ABSTRACT: The 4977 bp deletion of mitochondrial DNA is a molecular marker of cell ageing, and some solid tumors. This study was to detect deletion of mitochondrial DNA 4977 bp in laryngeal squamous cell cancer, and explore its correlation with differentiation of laryngeal cancer tissues.
Thirty-five specimens were collected from patients with pathologically diagnosed laryngeal squamous cell cancer. The total DNA was amplified to identify the 4977 bp deletion of mitochondrial DNA by polymerase chain reaction (PCR), and PCR products were verified by direct sequencing.
The 4977 bp deletion detected in 20 of 35 cancer tissues (57.14%), including 5 of 7 with well differentiation, 15 of 24 with moderate differentiation, and 0 of 4 with poor differentiation. Mutation incidence of cancer tissues with well or moderate differentiation were significantly higher than that of cancer tissues with poor differentiation, but there was no significant difference between cancer tissues with well and moderate differentiation.
The 4977 bp deletion of mitochondrial DNA was detected in laryngeal squamous cell cancer tissues, it might be relates to differentiation of cancer tissues.
No preview · Article · Nov 2004 · Ai zheng = Aizheng = Chinese journal of cancer