circRNA circ-CCND1 promotes the proliferation of laryngeal squamous cell carcinoma through elevating CCND1 expression via interacting with HuR and miR-646.

Cyclin D1 (CCND1) is a well-known proliferation promoter that accelerates G1/S transition in cancer. However, the underlying mechanism by which CCND1 is regulated is still largely unknown. In this study, we identified a novel circular RNA (circRNA) derived from CCND1 (circ-CCND1, hsa_circ_0023303) as a key regulator for CCND1. circ-CCND1 was found to be markedly up-regulated in laryngeal squamous cell carcinoma (LSCC) and closely associated with aggressive clinical features and adverse prognosis. Depletion of circ-CCND1 significantly inhibited LSCC cell proliferation in vitro and retarded tumour growth in vivo. Regarding the mechanism, circ-CCND1 physically bound to human antigen R (HuR) protein to enhance CCND1 mRNA stability; on the other hand, circ-CCND1 could act as an effective sponge for miR-646 to alleviate the repression of miR-646 on CCND1 mRNA. As a result, circ-CCND1 post-transcriptionally elevated CCND1 expression via coordinated avoidance of CCND1 mRNA decay, thereby promoting LSCC tumorigenesis. Taken together, our findings uncover the essential proliferation-promoting role of circ-CCND1 through regulation of the stability of CCND1 mRNA in LSCC. Targeting circ-CCND1 may be a promising treatment for LSCC patients.

Determination of antazoline hydrochloride in rat plasma and excreta by reversed-phase ion-pair chromatography and its application to pharmacokinetics.

A reversed-phase ion pair chromatography method with liquid-liquid extraction analytical method was developed and validated for the determination of antazoline hydrochloride in plasma and excreta of rat. The aim of our study was to characterize the preclinical pharmacokinetics and excretion profiles of antazoline hydrochloride in rats after intravenous injection at the dose of 10 mg/kg. Plasma and excreta samples were extracted with ethyl acetate, and phenacetin was used as the internal standard. The result showed that the method is suitable for the quantification of antazoline hydrochloride in plasma and excreta samples. Analysis of accuracy (90.89-112.33%), imprecision (<7.1%) and recovery (>82.5%) showed adequate values. After a single intravenous administration at 10 mg/kg to rats, plasma concentration profile showed a relative fast elimination proceeding with a terminal elimination half-life of 3.53 h. Approximately 61.8 and 14.2% of the administered dose were recovered in urine and bile after 72 and 24 h post-dosing respectively; 5.9% of the administered dose was recovered in feces after 72 h post-dosing. The above results show that the major elimination route is urinary excretion.

Ursolic acid ameliorates DNCB-induced atopic dermatitis-like symptoms in mice by regulating TLR4/NF-κB and Nrf2/HO-1 signaling pathways.

BACKGROUND: Ursolic acid (UA) is a triterpenoid compound found in natural plants. It has been reported to have anti-inflammatory, antioxidant, and immunomodulatory properties. However, its role in atopic dermatitis (AD) is unknown. This study aimed to evaluate the therapeutic effect of UA in AD mice and explore the underlying mechanisms. METHODS: Balb/c mice were treated with 2, 4-dinitrochlorobenzene (DNCB) to induce AD-like lesions. During modeling and medication administration, dermatitis scores and ear thickness were measured. Subsequently, histopathological changes, levels of T helper cytokines, and oxidative stress markers levels were evaluated. Immunohistochemistry staining was used to assess changes in the expression of the nuclear factor of kappa B (NF-κB) and NF erythroid 2-related factor 2 (Nrf2). Furthermore, CCK8 assay, reactive oxygen species (ROS) assay, real-time PCR, and western blotting were employed to evaluate the effects of UA on ROS levels, inflammatory mediator production, and the NF-κB and Nrf2 pathways in TNF-α/IFN-γ-stimulated HaCaT cells. RESULTS: The results showed that UA significantly reduced dermatitis score and ear thickness, effectively inhibited skin proliferation and mast cell infiltration in AD mice, and decreased the expression level of T helper cytokines. Meanwhile, UA improved oxidative stress in AD mice by regulating lipid peroxidation and increasing the activity of antioxidant enzymes. In addition, UA inhibited ROS accumulation and chemokine secretion in TNF-α/IFN-γ-stimulated HaCaT cells. It might exert anti-dermatitis effects by inhibiting the TLR4/NF-κB pathway and activating the Nrf2/HO-1 pathway. CONCLUSION: Taken together, our results suggest that UA may have potential therapeutic effects on AD and could be further studied as a promising drug for AD treatment.

[Clinical application of orofacial myofunctional therapy combined with muscle functionnal appliance in postoperative rehabilitation of children with OSA].

Objective:To explore the clinical significance of orofacial myofunctional therapy combined with muscle functional appliance in postoperative rehabilitation of children with OSA. Methods:Sixty children were diagnosed as moderate-to-severe OSA with AHI≥5 and underwent adenoid and/or tonsillar surgery. Children were divided into two groups based on whether they were willing to receive orofacial myofunctional therapy and muscle functional appliance after surgery. Lateral cephalogram and portable polysomnography were performed, and the pediatric OSA-18 scale was filled under the guidance of medical staff. The treatment group received combined treatment with orofacial myofunctional therapy and muscle functional appliance. Results:①General condition and subjective symptoms: The total score of OSA-18 in the treatment group was 65.15±11.25 preoperatively and 49.83±7.09 1-month postoperatively, while the score in the control group was 64.69±10.23 preoperatively and 48.07±6.87 1-month postoperatively. The results showed that postoperative sleep, physical symptoms, emotional status, daytime lethargy and energy status of patients, and their influence on their guardians were significantly improved in both groups（P<0.01）. The improvement of sleep disturbance, physical condition, daytime lethargy, the influence on their guardians were greater in the treatment group than in the control group 6-month and 12-month post-operatively（P<0.05）. These findings suggested that oral and facial muscle functional training combined with muscle functional appliance can provide greater improvement in general condition and subjective symptoms in the treatment group. ②PSG: Postoperative AHI and OAI were significantly decreased in both groups, while LSaO2 was significantly increased （P<0.01）, indicating that sleep respiratory function was significantly improved in both groups after treatment. Patients in treatment group showed greater AHI reduction and LSaO2 improvement 6-month and 12-month postoperatively（P<0.01）, indicating that oral and facial muscle functional training combined with muscle functional appliance can provide greater improvement in airway obstruction symptoms and sleep respiration. ③Radiological changes: SNB Angle was increased（P<0.05） and ANB Angle was decreased significantly（P<0.05）, while SPP-SPPW, U-MPW and TB-TPPW increased significantly in airway measurement 6-month and 12-month postoperatively （P<0.01）, indicating that after combined treatment with oral muscle functional training and muscle functional appliance, the mandible was moved forward and rotated clockwise. Conclusion:The combined treatment with oral muscle functional training and muscle functional appliance is more effective in improving oral breathing, upper airway sagittal structure and sleep breathing, and can correct oral habits of children. The long-term effect needs further investigation.

Long non-coding RNA CCAT2 drives the growth of laryngeal squamous cell carcinoma via regulating YAP activity.

Emerging evidence suggests that long non-coding RNA (lncRNA) is closely associated with numerous human diseases, including cancer. However, the functional relevance of lncRNA in human laryngeal squamous cell carcinoma (LSCC) is largely unknown. In the current study, we described CCAT2, a previously unappreciated oncogenic lncRNA in LSCC. CCAT2 was significantly upregulated in human LSCC tissue and serum samples, associated with larger tumor volume, higher clinical stage, and poorer differentiation status. Lentivirus-mediated CCAT2 knockdown notably repressed the cell viability, colony formation, and DNA synthesis rate of LSCC. Screening of transcription factors revealed that YAP/TEAD activity was affected by CCAT2 in LSCC cells. Further, CCAT2 directly binds to YAP protein and blocks the phosphorylation of YAP induced by LATS1, resulting in the nuclear translocation of YAP and the activation of YAP oncogenic targets, such as CTGF, CYR61 and AMOTL2. Importantly, we also confirmed the regulation of CCAT2 on YAP activity in vivo based on nude mice model. Altogether, we identified a novel lncRNA that controls YAP nucleocytoplasmic shuttling and promotes LSCC cell proliferation. Given the importance of YAP in tumorigenesis and progression, our results provide insights to intervene LSCC by targeting the CCAT2/YAP axis.

miR-181a and miR-203 inhibit migration and invasion of laryngeal carcinoma cells by interacting with ATF2.

MicroRNAs (miRNAs) have been recognized to modulate the progression of tumorigenesis by serving as oncogenes or tumor suppressors. Despite the involvement of miR-181a and miR-203 in several cancers as has been substantiated, their roles in laryngeal carcinoma (LC) remain unclear. In this study, the abundances of miR-181a, miR-203 and activating transcription factor 2 (ATF2) mRNA in LC cell lines were detected by RT-qPCR. Western blot was performed to detect the protein levels of N-cadherin, E-cadherin and ATF2. Cell migration and invasion ability were assessed by Trans-well assay. The putative binding sites between miR-181a or miR-203 and ATF2 were predicted using Bioinformatics software and further validated by Dual-Luciferase reporter and RNA immunoprecipitation (RIP) assays. Results showed reduced abundances of miR-181a and miR-203 in LC cell lines. Introduction of miR-181a or miR-203 reduced cell migration and invasion, which was further confirmed by the reduction of N-cadherin and increase of E-cadherin in LC cells. ATF2 was identified to be a potential target of miR-181a and miR-203. Absence of ATF2 overturned the stimulatory effects of anti-miR-181a and anti-miR-203 on cell migration and invasion in LC cells. Our findings suggested that miR-181a and miR-203 attenuated cell migration and invasion ability by directly targeting ATF2 in LC, providing novel insight into the regulatory mechanisms of miR-181a and miR-203 in LC.

Overexpression of Beclin1 inhibits proliferation and promotes apoptosis of human laryngeal squamous carcinoma cell Hep-2.

OBJECTIVE: Beclin1 was previously found to be downregulated in human laryngeal cancer (LC) tissues, and it results in poor prognosis. This study aimed to further confirm the antitumor effects of Beclin1 in LC cell line Hep-2. MATERIALS AND METHODS: Beclin 1 was overexpressed in Hep-2 cells using liposomal transfection and confirmed using reverse transcription polymerase chain reaction and Western blotting. Then, cell proliferation and apoptosis were determined in control (untransfected), empty vector transfected, and Beclin1 overexpressed groups using MTT and flow cytometry procedure, respectively. RESULTS: The expression of the Beclin1 gene in Hep-2 cells was significantly increased after vector transfection compared with control (1.173±0.046 vs 0.453±0.016, P<0.01) and empty vector (1.173±0.046 vs 0.440±0.021, P<0.01). Overexpression of Beclin1 inhibited proliferation at 4 days (0.619±0.051 vs 0.891±0.081 and 0.619±0.051 vs 0.878±0.105, P<0.01), 5 days (0.684±0.078 vs 1.127±0.094 and 0.684±0.078 vs 1.162±0.117, P<0.01), and 6 days (0.725±0.069 vs 1.168±0.103 and 0.725±0.069 vs 1.194±0.097, P<0.01) and promoted apoptosis (14.48%±1.42% vs 4.07%±0.66% and 14.48%±1.42% vs 4.39%±0.80%, P<0.01) in Hep-2 cells in comparison with the control and empty vector groups, respectively. CONCLUSION: Beclin1 may be an underlying target for the treatment of LC. This study has provided some experimental basis for the gene therapy of LC.

miR-200a-3p promotes the proliferation of human esophageal cancer cells by post-transcriptionally regulating cytoplasmic collapsin response mediator protein-1.

The dysregulation of cytoplasmic collapsin response mediator protein 1 (CRMP1) has been reported in lung cancer, medulloblastoma and esophageal cancer. However, the role of CRMP1 and its regulatory mechanisms in esophageal cancer remain unclear. In this study, we demonstrated that CRMP1 expression was downregulated in esophageal cancer tissues and that there were differences in its expression levels in different esophageal cancer cell lines. We found that CRMP1 overexpression inhibited the proliferation of esophageal cancer cells, whereas the silencing of CRMP1 promoted cell proliferation. We performed an analysis of potential microRNA (miRNA or miR) target sites using a commonly used prediction algorithm (TargetScan). The algorithm predicted that miR­200a-3p targets the 3' untranslated region (3'UTR) of CRMP1. Further experiments confirmed this prediction. In addition, we found that miR­200a-3p promoted the proliferation of esophageal cancer cells. Thus, our findings indicate that miR­200a-3p promotes the proliferation of human esophageal cancer cells by post-transcriptionally regulating CRMP1.

[A case of parapharyngeal space infection followed abdominal pain].

Descending necrotizing mediastinitis that has an abdominal pain as a main clinical manifestation is seldom. Here one case is reported. At the beginning, the patient had pharyngalgia and his swallowing was not smooth. After that, abdominal pain became a main symptom. Pharyngalgia relieved . However CT showed mediastinal infection. Surgical drainage,antibiotics treatment and nutritional support were performed. The patient was cured.

Pharmacokinetic study of Guanfu base G in rats by LC-ESI-MS.

A sensitive and simple liquid chromatography -: electrospray ionization mass spectrometry method has been established and validated for the quantification of Guanfu base G in rats. Phenoprolamine hydrochloride was selected as the internal standard. Sample preparation involved simple liquid-liquid extraction by ethylacetate with high efficiency. The chromatographical separation was performed on a Shimadzu C18 column (150 × 2.0 mm, 5 µm) with a gradient elution of 0.2% acetic acid-acetonitrile (30:70, v/v). The method was sensitive with the lowest limit of detection at 1 ng/mL (S/N ≥ 3) in 100 µL of rat plasma. Good linearity (r = 0.9996) was obtained covering a concentration of 5-2000 ng/mL. The intra- and interday assay precision ranged from 4.3 to 6.1% and 5.4 to 8.3%, respectively. In addition, the stability, extraction recovery and matrix effect involved in the method were also validated. After intravenous dosing, rat plasma Guangfu base G (GFG) concentration declined in a biphasic manner with a terminal elimination half-life of 3.72 h. The total plasma clearance values were 1.15 L/h/kg. After oral dosing, the plasma GFG concentration reached a maximum within 0.5 h. The absolute bioavailability of GFG was 83.06%.

[Expression of microtubule-associated protein 1 light chain 3 in laryngeal carcinoma].

OBJECTIVE: To investigate the expression of microtuble-associated protein 1 light chain 3 (LC3) in laryngeal squamous cell carcinoma (LSCC). METHOD: The expression of LC3 in 50 cases of LSCC, 45 cases of para-carcinoma, 10 cases of laryngeal papilloma and 16 cases of polyp of vocal cord were detected by immunohistochemistry (MaxVision method). Expression level of LC3 mRNA was assayed by RT-PCR in 41 of LSCC, 41 of para-carcinoma tissue and 11 of polyp of vocal cord. RESULT: The positive rates of LC3 protein expression were 60.0%, 93.3%, 90.0%, 93.8% in LSCC tissue, para-carcinoma, laryngeal papilloma and poly of vocal card tissues, respectively. The positive rates of LC3 were significantly lower in LSCC than in para-carcinoma and poly of vocal cord (chi2 = 18.135, P < 0.01). The mRNA levels of LC3 were significantly lower in LSCC than in para-carcinoma and poly of vocal cord (0.57 +/- 0.08 )vs (0.99 +/- 0.11) and (1.07 +/- 0.05) , F = -255.872, P < 0.01. The expression of LC3 were related to tumor location and pathological grade (P < 0.05), but not related to age, T stage, clinical stage and lymphoid metastasis (P > 0.05). CONCLUSION: Expression of LC3 are down-regulated in LSCC. The change of autophagic capacity may play an important role in occurrence and development of LSCC.

[Treatment of specific foreign bodies in tracheobronchial].

OBJECTIVE: To improve effectiveness of diagnosis and therapy in patients with tracheobronchial specific foreign body. METHOD: Fourty-six cases with specific foreign bodies in the trachea and bronchi, admitted from March 1998 to June 2006, were analyzed retrospectively. RESULT: Forty-one of 46 the specific foreign bodies were successfully removed through bronchoscopy. Three cases were removed by chest surgery. Two cases were died. CONCLUSION: Suitable surgical instrument should be well selected and the best operation should be applied according to different conditions of the foreign bodies. Rigid bronchoscopy for removal of specific foreign bodies under general anesthesia is a main measure presently.

[Clinical analysis of 33 cases of neurilemmomas of the head and neck].

OBJECTIVE: To investigate the diagnosis and treatment of neurinoma of the head and neck. METHOD: The clinical data of 33 patients with neurinoma of the head and neck, from 1980 to 2004, were retrospectively reviewed and analysed. RESULT: The tumors of all the cases were totally resected. Thirty of thirty-three cases have been followed up. Twenty-seven cases were fully recovered without any serious complication. One case with malignant tumor were died. The other 2 cases were died of other diseases. CONCLUSION: Tumor resection is still the most effective treatment available for patients with neurilemmomas of the head and neck. Resection of tumor should be performed as soon as possible with careful protection of the function of the nerve. Extensive resection should be performed to treat malignant tumors.

[Endoscopic transnasal approach in surgical treatment of ethmoid sphenoid diseases].

OBJECTIVE: To study the surgical experience of endoscopic sinus surgery for the patients with sphenoid sinus disease. METHOD: Twenty-one patients of sphenoid sinus diseases were treated with endoscopy by nasal cavity and sphenoid sinus. RESULT: During 3 months to four years following up, 20 patients were free from disease postoperatively, 1 patient underwent reoperation because of recurrence. CONCLUSION: Nasal endoscope provide the clear visual field, and operation under nasal endoscope could less pain and minimize injury to normal tissue. Transnasal-sphenoidal endoscopic sinus surgery under local anesthesia seems to be a safe and valuable way.