Melatonin reduces IL-33 and TSLP expression in human nasal epithelial cells by scavenging ROS directly.

BACKGROUND: Chronic rhinosinusitis (CRS) is a chronic mucosal inflammation of the nasal cavity and sinuses. It is classified into CRS without nasal polyps and CRS with nasal polyps (CRSwNP). CRSwNP has high recurrence, especially CRSwNP with massive eosinophil infiltration which is mediated by type 2 inflammatory response. Melatonin is a hormone secreted by the pineal gland, it has powerful antioxidant and anti-inflammatory effects in addition to regulating biological rhythms. There are no studies on melatonin for the treatment of CRS, so we aimed to explore whether melatonin could be used for the treatment of CRS. MATERIALS AND METHODS: In this study, we used melatonin to treat a cell model of CRS. Subsequently, MTT assay was performed to examine the cell viability of human nasal epithelial cells (HNEpCs), a reactive oxygen species (ROS) kit to detect ROS production, a malondialdehyde (MDA) kit to detect the MDA content in the cell culture supernatant, and an apoptosis kit and Western blot analysis to detect apoptosis. The expressions of Nrf2, HO-1, IL-33, TSLP, and IL-25 were detected by Western blot analysis. RESULTS: Melatonin improved the viability of HNEpCs, reduced lipopolysaccharide-induced ROS, reduced the MDA content, and inhibited their apoptosis. More importantly, melatonin reduced the expression of IL-33 and TSLP, an important phenomenon for the treatment of CRSwNP. CONCLUSION: Melatonin protects HNEpCs from damage in inflammation and reduces IL-33 and TSLP expression of HNEpCs.

HO-1: a new marker for predicting postoperative recurrence of CRSwNP.

BACKGROUND: Chronic rhinosinusitis with polyps (CRSwNP) is a subtype of chronic rhinosinusitis and is highly prone to recurrence; therefore, it is urgent to find appropriate markers to predict recurrence of CRSwNP after surgery. PURPOSE: We aim to investigate the expression of HO-1 in CRSwNP and assess its value of predicting postoperative recurrence of CRSwNP. METHODS: We recruited 77 participants and collected clinical data of all. We use Immunohistochemical staining to determine the expression of HO-1 in tissues. We use Spearman correlation test to analyze the correlation between HO-1 positive cell count and clinical score, and ROC curve to assess the value of HO-1 positive cell count in predicting recurrence of CRSwNP. RESULTS: HO-1 positive cells were macrophages and significantly increased in CRSwNP; HO-1 positive cell count was negatively correlated with preoperative SNOT-22 score; HO-1 can predict postoperative recurrence of CRSwNP, AUC = 0.80, p = 0.004. CONCLUSION: HO-1 is a biochemical marker of CRSwNP and can predict postoperative recurrence of CRSwNP.

RP11­156L14.1 regulates SSR1 expression by competitively binding to miR­548ao­3p in hypopharyngeal squamous cell carcinoma.

Emerging studies have demonstrated that long non­coding RNAs (lncRNAs) play essential roles in tumorigenesis. However, the role and function of lncRNAs in hypopharyngeal squamous cell carcinoma (HSCC) have not been completely elucidated. The present study explored the function of a novel lncRNA, RP11­156L14.1, in HSCC. RP11­156L14.1 was revealed to be highly expressed in HSCC tissues and cell lines. Knockdown of RP11­156L14.1 inhibited proliferation, migration, and invasion in HSCC cells. Furthermore, RP11­156L14.1 regulated epithelial­mesenchymal transition (EMT) by controlling EMT­related protein expression. Mechanistically, RP11­156L14.1 exerted its function as a competing endogenous RNA (ceRNA) and directly interacted with miR­548ao­3p. The present study also demonstrated that miR­548ao­3p regulated signal sequence receptor subunit 1 (SSR1) expression by targeting SSR1 3'­UTR. Moreover, the xenograft HSCC tumor model revealed that knockdown of RP11­156L14.1 markedly suppressed HSCC tumor growth in vivo. In summary, these findings indicated that the lncRNA RP11­156L14.1 functions as an oncogene in HSCC by competing with miR­548ao­3p in regulating SSR1 expression. The RP11­156L14.1/miR­548ao­3p/SSR1 axis could be utilized as a potential novel biomarker and therapeutic target for HSCC.

[Construction of a recombinant eukaryotic vector of Atoh1 and its expression in 293-T cells].

OBJECTIVE: To clone the coding sequence of Atoh1 cDNA from SD rat and construct a eukaryotic expression vector for its expression in eukaryotic cells. METHODS: The total RNA was extracted from colonic mucosa of SD rats and the double-stranded cDNA of Atoh1 was amplified using RT-PCR. The cDNA coding sequence was then cloned into PMD-19T vector followed by sequence analysis. The digested fragment, after purification, was linked into the eukaryotic expression vector pIRES2-EGFP containing the EGFP gene and the internal ribosomes site (IRES). After identification by enzyme digestion and sequence analysis, the recombinant plasmid was transfected into 293T cells via Lipofectamine, and the expression of the target protein was detected under fluorescence microscope, using PT-PCR and Western blotting. RESULTS: DNA sequence analysis showed that the amplified rat Atoh1 gene, with a length of 1056 bp of the coding sequence which encoded 351 amino acids, had two base mutations compared to the reference sequences in GenBank, possibly as a result of single nucleotide polymorphisms that induced nonsense mutation without affecting the amino acid sequences or the protein expression. The results of enzyme digestion and sequence analysis demonstrated that the Atoh1 gene was correctly inserted in the eukaryotic expression vector plRES2-EGFP. Fluorescence microscopy and Western blotting both confirmed successful expression of Atohl at the mRNA and protein levels in 293T cells 48 h after transfection with the recombinant plasmid. CONCLUSION: The recombinant plasmid harboring the coding sequence of SD rat Atoh1 gene has been constructed successfully and can be expressed in the 293T cells, which provides a basis for functional study of Atoh1 gene and future researches in gene therapy for sensorineural hearing loss.

[Long term follow up of midline partial glossectomy and uvulopalatopharyngoplasty for treating obstructive sleep apnea hypopnea syndrome].

OBJECTIVE: To observe the long-term results of midline partial glossectomy with uvulopalatopharyngoplasty (UPPP) in the treatment of obstructive sleep apnea hypopnea syndrome (OSAHS). METHODS: Twenty-four severe OSAHS patients treated with midline partial glossectomy and UPPP from January 2003 to March 2004 were included in this study, the follow-up was 5 years. The median of preoperative lowest arterial oxygen saturation (LSaO(2)) of this group at night (the same below) 0.650, and AHI was 56.5 times/h, UPPP was performed under general anesthesia, no tracheotomy performed. Criteria of curative effects: AHI < 5 times/h was recovery, AHI < 20 times/h and decreased beyond 50% marked improvement, only AHI decreased beyond 50% improvement. RESULTS: Post-operation AHI (6 months, 1 year, 2 years and 5 years after surgery) decreased significantly compared to that before the surgery, and post-operation LSaO(2) was significantly higher than that of preoperative (Wilcoxon's signed rank test, the same below, P < 0.01). The LSaO(2) and AHI were significantly different between 1 year, 2 years, 5 years and 6 months post-operatively (P < 0.01). Six months after surgery, PSG results showed that 21 were recovery, marked improvement for the other 3 cases, the recovery rate was 87.5%. One year after surgery, 18 were recovery, marked improvement in 3 cases, the recovery rate 75.0%. Two years after surgery, 14 cases recovery, marked improvement in 4 cases, the recovery rate 58.3%. Five years after surgery, 6 were recovery, the recovery rate 25.0%. Among 5 cases with hypertension before the surgery, after surgery antihypertensive drugs were not necessary in 4 cases, and the dosage was decreased in 1 case. CONCLUSION: The midline partial glossectomy with UPPP surgery may be an effective treatment for the severe OSAHS, long-term effect is satisfactory.

[Growth retardation in children with obstructive sleep apnea hypopnea syndrome].

OBJECTIVE: To observe changes of growth, body composition and biochemical markers associated with growth (IGF-1) in prepubertal children with obstructive sleep apnea syndrome (OSAHS). METHODS: Thirty-one children aged 3-10 years with OSAHS were followed up for 1 year after the corresponding surgery. During the same period of time, 20 children of similar age without OSAHS (excluded any other diseases that could result growth retardation or hypoxemia) were also followed up for 1 year. PSG, height, weight as well as insulin-like growth factor-1 (IGF-1) were measured during the preoperative period, 3 months, 6 months and 1 year after surgery in patient group. The same indexes were measured before surgery, and only height and weight were recorded after surgery in the control group. Wilcoxon signed- rank test and Mann-Whitney U test are used to analyze the data. RESULTS: The lowest oxyhemoglobin saturation of the patient group (0.88) is significantly lower than that of the control group (0.98), and was found increased at the 6 months post-op follow up (0.97, U = 238.5, P > 0.05), no significant change was found at the 1 year follow up. The post-op AHI (6 months after surgery) of the patient group (6.0/h) decreased to the similar level of the control group (0/h, U = 240.0, P > 0.05), and was similar to 1 year after surgery. Height of the patient group (116 cm), which was lower than the control group (U = 127.0, P < 0.001), significantly increased 1 year (138 cm) after the corresponding surgery (Z = 3.726, P < 0.01), and reached the similar levels of the control group (137 cm) 1 year after the surgery (U = 123.5, P > 0.05). The serum IGF-1 levels of the patient group (33.7 ng/ml), which were significantly lower than those of the controls preoperatively (44.1 ng/ml, U = 206.0, P < 0.05), increased to similar levels with the controls 6 months after the operation (50.3 ng/ml, U = 261.0, P > 0.05), and the 1 year post-op follow up was similar to the control group too (48.6 ng/ml, U = 163.0, P > 0.05). CONCLUSIONS: The cure of OSAHS could accelerate growth in prepubertal children, and the serum IGF-1 levels increases at the same time. The growth retardation is presumed in children with OSAHS.

[Sediment record of phosphorus and the primary study of its bioavailability in Jiaozhou Bay sediments].

The geochemical characteristics of phosphorus forms in different grain size sediments, the influencing factors as well as the bioavailability of phosphorus in Jiaozhou Bay were investigated employing the sequential extraction method. The results showed that inorganic phosphorus was the dominant form of total phosphorus and organic phosphorus was only the minor part. The detrital carbonate-bound phosphorus was the largest part of total phosphorus. The grain size, organic carbon, temperature, pH and etc. are the main controlling factors of phosphorus in sediments. Except for Ca-P, most of other phosphorus forms contents increased as the grain size decreased. The bioavailability research showed that the bioavailable phosphorus was mainly the exchangeable phosphorus, phosphorus bound to Al and Fe and organic phosphorus. Moreover, the potential bioavailable phosphorus proportion in total phosphorus increased as the grain size decreased. The bioavailable phosphorus in different grain size sediments has positive correlationship with phytoplankton abundance and phosphate in overlying water.

[Theory of local inflammatory infiltration and irritation in the pathogenesis of acquired cholesteatoma].

OBJECTIVE: To explore the deep pathogenesis of acquired cholesteatoma. METHODS: The temporal bone slides of 12 ears with retraction pocket were histopathologically studied under microscope, especially focusing on the location of retraction pocket and inflammatory pathology in the local middle ear cavity next to retraction pockets. The temporal bone slides of 11 ears with acquired cholesteatoma were histopathologically observed and 33 cases diagnosed as acquired cholesteatoma were clinically observed observed in the local middle ear cavity next to the part without retraction pocket of eardrum. The results of pathological observation of the temporal bone slides with acquired cholesteatoma and clinical observation during operation for acquired cholesteatoma show that cholesteatoma invade mainly and occupied the ossicular chain eara of the middle ear cleft. CONCLUSION: In the pathological process of otitis media, the intractable pathological changes in the ossicular chain area can inward adhere posterosuperior quadrant or pars flaccida of the eardrum to form retraction pocket and permanently infiltrate the external squamous epithelial layer of retraction pocket to excessively proliferate and keratinize, leading to formation of acquired cholesteatoma.