Changes in percentage of GATA3+ regulatory T cells and their pathogenic roles in allergic rhinitis / 南方医科大学学报

OBJECTIVE@#To investigate the changes in percentage of GATA3+ regulatory T (Treg) cells in patients with allergic rhinitis (AR) and mouse models.@*METHODS@#The nasal mucosa specimens were obtained from 6 AR patients and 6 control patients for detection of nasal mucosal inflammation. Peripheral blood mononuclear cells (PBMC) were collected from 12 AP patients and 12 control patients to determine the percentages of Treg cells and GATA3+ Treg cells. In a C57BL/6 mouse model of AR, the AR symptom score, peripheral blood OVA-sIgE level, and nasal mucosal inflammation were assessed, and the spleen of mice was collected for detecting the percentages of Treg cells and GATA3+ Treg cells and the expressions of Th2 cytokines.@*RESULTS@#Compared with the control patients, AR patients showed significantly increased eosinophil infiltration and goblet cell proliferation in the nasal mucosa (P < 0.01) and decreased percentages of Treg cells and GATA3+ Treg cells (P < 0.05). The mouse models of AR also had more obvious allergic symptoms, significantly increased OVA-sIgE level in peripheral blood, eosinophil infiltration and goblet cell hyperplasia (P < 0.01), markedly lowered percentages of Treg cells and GATA3+ Treg cells in the spleen (P < 0.01), and increased expressions of IL-4, IL-6 and IL-10 (P < 0.05).@*CONCLUSION@#The percentage of GATA3+ Treg cells is decreased in AR patients and mouse models. GATA3+ Treg cells possibly participate in Th2 cell immune response, both of which are involved in the occurrence and progression of AR, suggesting the potential of GATA3+ Treg cells as a new therapeutic target for AR.

The expression and significance of TRPM8 among chronic rhinosinusitis with nasal polyps / 中华耳鼻咽喉头颈外科杂志

Objective: To compare the expression and difference of melastatin-related transient receptor potential 8(TRPM8) among chronic rhinosinusitis, nasal polyps and normal mucosa tissues. And to explore the significant expression of TRPM8 among CRSwNP. Methods: Fifty-one patients underwent endoscopic sinus surgery in the Department of Otorhinolaryngology Head and Neck Surgery of Renmin Hospital of Wuhan University from February 2019 to January 2020 were recruited, including 33 males and 18 females, aged from 14 to 65 years old (34.55±1.689).Immunohistochemistry was used to detected the expression of TRPM8 protein among CRSsNP(17),CRSwNP (17) and control tissuses(17). In addition, the correlation between the expression of TRPM8 protein in CRSwNP patients and preoperative CT Lund-Mackay scores and preoperative VAS scores and sinonasal outcome test-20 scores was analyzed, respectively. The primary human nasal epithelial cells were cultured in vitro and the expression of TRPM8 was detected by quantitative real-time PCR and western blotting . The tissue in control group, chronic rhinosinusitis without nasal polyps (CRSsNP) group and the CRSwNP group were collected and grinded into tissue homogenized. The expression of TRPM8 protein was detected by western blotting after 24 h stimulation after homogenate was added into the medium of RPMI 2650 and primary nasal epithelial cells. Results: Compared with the control, the expression of TRPM8 was significantly up-regulated in nasal polyps (t=6.852, P<0.05). TRPM8 was mainly expressed in epithelial cells. The expression of TRPM8 in the epithelial cells of CRSsNP had no difference with the control group (t=1.980, P>0.05). In addition, the expression of TRPM8 in CRSwNP patients was positively correlated with the preoperative CT Lund-Mackay scores and VAS scores and SNOT-20 scores (r=0.512, P<0.05;r=0.853, P<0.01;r=0.814, P<0.01). After cultured primary epithelial cells in vitro, the expression level of TRPM8 in epithelial cells derived from nasal polyp was significantly higher than that in control group (t=8.845, P<0.05). By adding the homogenization of control and CRSsNP and CRSwNP tissues, the expression of TRPM8 in RPMI 2650 cells and primary nasal epithelial cells was changed and that was significantly increased after adding the homogenization of the group of CRSwNP. Conclusion: TRPM8 is highly expressed in nasal polyps epithelial cells, suggesting that TRPM8 may be involved in the pathogenesis of nasal polyps regulated by nasal epithelial cells.

Influence of Leukodeplated Blood Transfusion on Cellular Immunity of Acute Leukemia Patients / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the influence of leukodeplated blood transfusion on cellular immunity of patients with acute leuemia, so as to provide support for application of leuko-deplated blood transfusion in clinic.</p><p><b>METHODS</b>A total of 100 AL patients from January 2012 to December 2015 were chosen, and were divided into 2 groups: leukodeplated blood transfusion group(50 cases) and routine blood transfusion group(RBT) as control (50 cases). The effective rate, side effects, peripheral blood T cells and expression level of TLR2 and TLR4 were compared between 2 groups.</p><p><b>RESULTS</b>The expression levels CD3(+), CD4(+), CD8(+), CD4(+)/CD8(+) of TLR2 and TLR4 in control group were (52.18±2.14)%, （27.28±1.19）%,（24.21±1.65）%,1.22±0.18,0.62±0.04 and 0.57±0.05, respectively, after treatment; while these indicators in LdBT group were （52.18±2.14）%,（30.97±2.01）%,（27.08±1.55）%,1.39±0.24,0.91±0.06 and 0.87±0.07, respectively, and above-mentioned indicators in LdBT group were significantly higher than those in control group（P<0.05）. Compared with these indicators before treatment, CD3(+), CD4(+), CD8(+) and CD4(+)/CD8(+) in the patients increased significantly（P<0.05）. The efficiency was 92.00% (46/50) in LdBT group, and 84.00% (42/50) in control group, without statistically significant difference（P>0.05）. The rate of side effects in study group was 6% (3/50), 18% (9/50) in control group, with statistically significance difference （P<0.05）.</p><p><b>CONCLUSION</b>Leukodeplated blood transfusion can improve the cellular immunity of AL patients, and reduce the rate of side effects.</p>

Comparison of outcomes between endoscopic surgery and conventional nasal packing for epistaxis in the posterior fornix of the inferior nasal meatus

Objective: To investigate the clinical features of epistaxis in the posterior fornix of the inferior nasal meatus and compare the treatment outcomes of endoscopic surgery and conventional nasal packing for this intractable form of epistaxis

Methods: Between August 2011 and August 2014, the medical records of 53 adult patients with idiopathic epistaxis in the posterior fornix of the inferior nasal meatus diagnosed by nasal endoscopy were obtained from our department. Of these, 38 patients underwent endoscopic surgery [surgery group] and 15 received a nasal pack [packing group]. The patients' background characteristics, incidence of re-bleeding, extent of discomfort after treatment as assessed using a 10-point visual analogue scale [VAS] and incidence of nasal cavity adhesion after treatment were analysed

Results: There were no significant differences in background characteristics between the two groups. The incidence of re-bleeding [0/38 vs. 4/15, surgery vs. control, P = 0.001], VAS score for discomfort [2.4 +/- 1.4 vs. 7.6 +/- 1.0, surgery vs. control, P = 0.001] and incidence of nasal cavity adhesion after treatment [2/38 vs. 7/15, surgery vs. control, P = 0.007] were significantly lower in the surgery group than in the packing group

Conclusion: Endoscopic surgery is superior to conventional nasal packing for the management of epistaxis in the posterior fornix of the inferior nasal meatus. During surgery, it is crucial to expose the bleeding sites by shifting the inferior turbinate inward by fracture

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>

Relationship between bacterial biofilm and chronic rhinosinusitis: a systematic review / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the relationship between bacterial biofilm (BBF) and chronic rhinosinusitis (CRS).</p><p><b>METHODS</b>The database on line was searched to collect the studies on BBF and CRS. The method of meta analysis was used to analyze the data of suitable studies.</p><p><b>RESULTS</b>Fourteen studies were included. System evaluation indicated that the BBF detection rate in CRS group was significantly higher than that in the control group (OR = 17.01, P < 0.01), and the nasal surgery's rate of BBF positive group was significantly higher than the negative group (OR = 3.99, P < 0.01). Preoperative Lund-Kennedy endoscopic score, Lund-MacKay CT score, symptom severity score, postoperative Lund-Kennedy score after six months showed no difference between BBF positive group and negative group.</p><p><b>CONCLUSION</b>The presence of BBF is related to the pathogenesis of CRS and the history of nasal surgery.</p>

Expressions of clinical significances of p-extracellular regulated kinase 1/2 and matrix metalloproteinase-9 in cervical squamous cell carcinoma / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the expressions and clinical significances of p-extracellular regulated kinase(P-ERK)1/2 and matrix metalloproteinase-9(MMP-9)in cervical squamous cell carcinoma.</p><p><b>METHODS</b>The expressions of P-ERK1/2 and MMP-9 in 30 cases with chronic cervicitis, 45 cases with cervical intraepithelial neoplasia (CIN), and 58 cases with cervical squamous cell carcinoma were detected by immunohistochemical method.</p><p><b>RESULTS</b>The positive rates of P-ERK1/2 and MMP-9 in chronic cervicitis, CIN, and cervical squamous cell carcinoma were 0 and 0, 28.9% and 24.4%, 77.6% and 65.5%, respectively, showing significant differences among these three groups (χ(2)= 54.393,p=0.003;χ(2)=40.968,p=0.005). The positive rates of P-ERK1/2 and MMP-9 in patients at clinical stages 2-3, at G3, with lymphatic metastasis, or with a tumor diameter greater than 4 cm were significantly higher than those at clinical stage 1(p=0.015,p=0.002), at G1-G2(p=0.013,p=0.017), without lymphatic metastasis (p=0.017,p=0.021), or with a tumor diameter less or equal than 4 cm in cervical squamous cell carcinoma(p=0.008,p=0.004). There was a positive correlation between P-ERK1/2 and MMP-9 in cervical squamous cell carcinoma (χ(2)=8.955,p=0.006).</p><p><b>CONCLUSIONS</b>The expressions of P-ERK1/2 and MMP-9 increase gradually with the progression of cervical squamous cell carcinoma. The over expressions of P-ERK1/2 and MMP-9 may promote the infiltration of cervical squamous cell carcinoma and lymphatic metastasis, druing which these two enzymes may exert their effects in a synergistic manner.</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>

Perioperative management of modified uvulopalatopharyngoplasty / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>Experiences and lessons of uvulopalatopharyngoplasty (UPPP ) perioperative management, especially causes of postoperative tracheotomy, were analyzed, and related strategy was raised to have a better perioperative management and to avoid tracheotomy.</p><p><b>METHODS</b>Two hundred and fifty eight cases of obstructive sleep apnea hypopnea syndromes (OSAHS) diagnosed with polysomnography (PSG) were treated with modified uvulopalatopharyngoplasty (UPPP). The perioperative management was summarized. Patients were divided into two groups according to the perioperative management: without or with perioperative comprehensive management. In group A, there were 32 patients, without comprehensive management, and in group B there were 226 cases with comprehensive management. Sixty eight cases in group B whose apnea hypopnea index over 50 times per hour and the lowest arterial oxygen saturation was less than 0.5 were treated with continuous positive airway pressure (CPAP) for 1 to 3 weeks. For all the 258 cases, perioperative management includes treatment of medical complications, treatment with antibiotics 2 or 3 days before the operation. None of these cases had tracheotomy before surgery.</p><p><b>RESULTS</b>In group A, three of 32 patients had postoperative tracheotomy, two because of bleeding, and another one because of laryngeal spasm. In group B, none of 226 patients underwent tracheotomy, which owing to modified operative apparatus and effective perioperative and postoperative treatment (chi2 = 21.35, P < 0.001). In group A, 5 of 32 patients had oral pharynx bleeding after 24 hours of the operation. While 26 of 226 patients in group B did so (chi2 = 0.15, P > 0.05).</p><p><b>CONCLUSION</b>Comprehensive perioperative management can effectively lower down the complication rate for patients receiving uvulopalatopharyngoplasty.</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>