CircRNA hsa_circ_0069,399 as a potential clinical prognostic marker in laryngeal squamous cell carcinoma.

Objective: Circular RNAs (circRNAs) significantly influence the invasion, metastasis, gene expression, proliferation, and apoptosis of tumor cells. However, the roles of circRNAs in laryngeal squamous cell carcinoma (LSCC) remain largely unexplored. This study aims to examine circRNA expression patterns in LSCC and adjacent non-tumorous tissues, with the goal of uncovering potential biomarkers for LSCC. Methods: Tissue samples were collected from both the tumor and adjacent normal tissues of ten patients who had undergone surgical resection. The profiling of circRNAs was conducted through transcriptomic sequencing and analytical bioinformatics approaches. A ternary regulatory network based on the competitive endogenous RNA (ceRNA) hypothesis was established, linking target circRNAs to clinical immunohistochemical parameters for comparison. Verification of target circRNAs in LSCC tissues was performed using quantitative real-time PCR (RT-qPCR), whereas target mRNAs were analyzed through immunohistochemistry. Results: A total of 126 significantly different circRNAs were identified, including 40 up-regulated genes and 86 down-regulated genes. Furthermore, 92 circRNA-miRNA-mRNA regulatory relationship axes related to clinical immunohistochemical indicators were found based on 5 candidate circRNAs. Interestingly, all axes related to the target genes MKI67 and TP53 were found to compete with the same circRNA: hsa_circ_0069,399. Further verification confirmed that the hsa_circ_0069,399 expression was overtly upregulated in tumor tissues from LSCC patients, which was consistent with the sequencing results. Conclusion: hsa_circ_0069,399 could be a potential prognostic marker for LSCC.

NRSN2 promotes the malignant behavior of HPV-transfected laryngeal carcinoma cells through AMPK/ULK1 pathway mediated autophagy activation.

Neurensin-2 (NRSN2) performs a pro-carcinogenic function in multiple cancers. However, the function of NRSN2 in HPV-infected laryngeal carcinoma (LC) remains unclear. HPV transfection was performed in LC cells. The mRNA and protein levels were monitored using RT-qPCR, immunoblotting, and IF. Cell viability and proliferation were found using the CCK-8 assay and Edu staining. Cell invasion, migration, and apoptosis were probed using the Transwell, wound healing, and flow cytometry, respectively. The autophagosome was observed using TEM. NRSN2 was overexpressed in HPV-transfected LC cells. Inhibition of NRSN2 restrained the autophagy and malignant behavior of HPV-transfected LC cells. Meanwhile, the inhibition of AMPK/ULK1 pathway limited the increased autophagy of HPV-transfected LC cells caused by NRSN2 overexpression. Furthermore, NRSN2 knockdown inhibits autophagy by suppressing AMPK/ULK1 pathway, thereby restraining the malignant behavior of HPV-transfected LC cells. Our research confirmed that HPV transfection increased the autophagy and malignant behavior of LC cells by regulating the NRSN2-mediated activation of the AMPK/ULK1 pathway, offering a new target for cure of LC.

Correlation analysis of TGF-ß1, MMP-9, TIMP-1, IL-1, IL-4, IL-6, IL-17, and TNF-α in refractory chronic rhinosinusitis: A retrospective study.

OBJECTIVE: To investigate the potential correlation of transforming growth factor-ß (TGF-ß), matrix metalloprotein 9 (MMP-9), tissue inhibitor of metalloproteinases 1 (TIMP-1), Interleukin 1 (IL-1), IL-4, IL-6, IL-17, and tumor necrosis factor alpha (TNF-α) in refractory chronic rhinosinusitis. METHODS: A total of 150 participants were retrospectively included in this study from August 2018 to February 2020. The people enrolled were equally allocated into refractory group (patients with refractory chronic rhinosinusitis), chronic group (patients with chronic rhinosinusitis), and control group (normal people). The level of TGF-ß1, MMP-9, TIMP-1, IL-1, IL-4, IL-6, IL-17, and TNF-α were recorded. The unconditional multivariate binary logistic regression was used to analyze the factors affecting refractory chronic rhinosinusitis. RESULTS: The Davos score, T&T olfactometer threshold test, and Lund-Mackay CT scores in refractory group were significantly higher than the chronic group (P<0.05). The level of TGF-ß1, MMP-9, TIMP-1, IL-1, IL-4, IL-6, IL-17, and TNF-α in the refractory group were significantly higher than the chronic group and the control group (all P<0.05). Similarly, the level of the above mentioned indexes in the chronic group were significantly higher than the control group (P<0.05). The Davos score, T&T olfactometer threshold test score, Lund-Mackay CT score, and the level of TGF-ß1, MMP-9, TIMP-1, IL-1, IL-4, IL-6, IL-17, and TNF-α positively correlated with refractory chronic rhinosinusitis. Moreover, the unconditional multivariate binary logistic regression showed that the influencing factors of refractory chronic rhinosinusitis included TGF-ß1, MMP-9, TIMP-1, IL-1, IL-4, IL-6, IL-17, and TNF-α. CONCLUSION: The findings of the present study provide evidence for TGF-ß1, MMP-9, TIMP-1, IL-4, IL-6, IL-17, and TNF-α as the influencing factors of refractory chronic rhinosinusitis.

Association of Laryngopharyngeal Reflux Disease and Refractory Chronic Rhinosinusitis.

OBJECTIVE: The current study was conducted to explore the association between laryngopharyngeal reflux disease (LPRD) and refractory chronic rhinosinusitis (RCRS), and to reveal the potential role of LPRD in the development of RCRS. METHODS: A total of 104 patients diagnosed as chronic rhinosinusitis (CRS) and hospitalized, as well as 50 healthy participants were enrolled into this study. Among patients enrolled, 53 were RCRS and 51 were CRS. All participants were assessed with reflux symptom index (RSI) and reflux finding score (RFS). Patients with RSI >13 and/or RFS >7 were diagnosed as LPRD. In addition, pepsin in the nasal secretions from the middle meatus tract complex at fasting in the morning was analyzed using ELISA. Patients with pepsin concentration of >75 ng/ml was considered as positive, and confirmed as LPRD. RESULTS: There was no significant difference in age, sex, height, and weight among RCRS, CRS, and control groups. The positive rates of RSI and RFS scores were 39.6% in the RCRS and 52.9% in the CRS, respectively, which were significantly higher than that of the control group (14.0%, P < 0.05), respectively. Significantly higher positive rate of pepsin assay was also observed for RCRS(43.4%) and CRS(64.7%) when compared with the control group (18.0%, P < 0.05). The positive rate of pepsin assay was strongly associated with the positive rate of RSI or RFS (P < 0.001). CONCLUSION: The present findings have important implications that RSI and RFS score was highly associated with pepsin assay result in the patients with RCRS or CRS. We confirmed the importance of RSI and RFS scoring system as well as pepsin test, which may offer insight for clinical screening for LPRD among CRS patients.

Regulatory Role of Zinc in Allergic Rhinitis through the IL-33/ST2 Pathway.

OBJECTIVE: We aimed to investigate the expression of serum zinc and cytokines interleukin- (IL-) 13 and IL-33 in patients with allergic rhinitis (AR) and observe the effects of zinc on cytokines and pathway proteins in P815 mast cells stimulated by Artemisia annua allergen (Art.) in the IL-33/suppression of the tumorigenicity 2 (ST2) pathway. We also aimed to explore the possible regulatory role of zinc in AR and provide new ideas to determine the etiology and treatment of AR. METHODS: AR patients treated from March to September in 2018 were selected as the research participants, and 50 healthy people in the same period were selected as the control group. Serum samples of all patients were collected, and those of AR patients were tested for the presence of allergens. The expression of IL-13 and IL-33 was detected by performing an enzyme-linked immunosorbent assay, while the serum zinc level was detected by conducting an inductively coupled plasma mass spectrometry. The cell counting kit (CCK-8) was used to detect the proliferation of P815 mast cells, and western blot was used to detect the expression of ST2, p38, and p65 proteins. RESULTS: A total of 92 AR patients were included in the study; of them, 52 had mild AR, while 40 had moderate AR. The primary allergen found in AR patients was Artemisia, and the positivity rate was 53.26%. The serum zinc ion level of AR patients decreased, and the expression of IL-13 and IL-33 increased. After Art. was used to treat P815 mast cells, the expression of IL-33 in the cell supernatant increased in a concentration-dependent manner, the expression of receptor ST2 increased, and the expression of downstream p38 and p65 proteins increased. However, after treatment with ZnSO4, the expression of IL-33 in the cell supernatant decreased, and the expression of ST2, p38, and p65 protein decreased. CONCLUSION: The serum zinc level of AR patients decreased. In the IL-33/ST2 pathway, ZnSO4 can reduce the hypersensitivity of mast cells induced by Art.

Traditional Chinese medicines and their active ingredients sensitize cancer cells to TRAIL-induced apoptosis.

The rapidly developing resistance of cancers to chemotherapy agents and the severe cytotoxicity of such agents to normal cells are major stumbling blocks in current cancer treatments. Most current chemotherapy agents have significant cytotoxicity, which leads to devastating adverse effects and results in a substandard quality of life, including increased daily morbidity and premature mortality. The death receptor of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can sidestep p53-dependent pathways to induce tumor cell apoptosis without damaging most normal cells. However, various cancer cells can develop resistance to TRAIL-induced apoptosis via different pathways. Therefore, it is critical to find an efficient TRAIL sensitizer to reverse the resistance of tumor cells to TRAIL, and to reinforce TRAIL's ability to induce tumor cell apoptosis. In recent years, traditional Chinese medicines and their active ingredients have shown great potential to trigger apoptotic cell death in TRAIL-resistant cancer cell lines. This review aims to collate information about Chinese medicines that can effectively reverse the resistance of tumor cells to TRAIL and enhance TRAIL's ability to induce apoptosis. We explore the therapeutic potential of TRAIL and provide new ideas for the development of TRAIL therapy and the generation of new anti-cancer drugs for human cancer treatment. This study involved an extensive review of studies obtained from literature searches of electronic databases such as Google Scholar and PubMed. "TRAIL sensitize" and "Chinese medicine" were the search keywords. We then isolated newly published studies on the mechanisms of TRAIL-induced apoptosis. The name of each plant was validated using certified databases such as The Plant List. This study indicates that TRAIL can be combined with different Chinese medicine components through intrinsic or extrinsic pathways to promote cancer cell apoptosis. It also demonstrates that the active ingredients of traditional Chinese medicines enhance the sensitivity of cancer cells to TRAIL-mediated apoptosis. This provides useful information regarding traditional Chinese medicine treatment, the development of TRAIL-based therapies, and the treatment of cancer.

Role of Zinc Sulphate in Immune Regulation in Artemisia annua Pollen-challenged P815 Mastocytoma Cells.

Objective This study aimed to investigate the role of zinc sulphate in immune regulation in Artemisia annua pollen-challenged P815 mastocytoma cells. Methods P815 mastocytoma cells were treated with various concentrations of zinc sulphate and Artemisia annua pollen. Cell proliferation was measured using the Cell Counting Kit-8. The amount of ST2 and p38 in the cells were measured using Western blotting. The level of interleukins (IL)-33 in the supernatant was determined using the enzyme-linked immunosorbent assay. The levels of IL-2, IL-4, IL-5, interferon-γ, and tumor necrosis factor were measured using the cytometric bead array. Results Artemisia annua pollen at a concentration >0.001 µg/mL induced allergic response in the P815 mastocytoma cells. Expressions of IL-33, IL-4, ST2, and p38 increased along with higher concentrations of Artemisia annua pollen. Zinc sulphate of 50-200 µmol/L promoted the proliferation of P815 mastocytoma cells. Zinc sulphate attenuated the upregulation of IL-33, IL-4, ST2, and p38 caused by Artemisia annua pollen. Conclusion Zinc sulphate can promote the proliferation of P815 mastocytoma cells. It can also attenuate allergic response in the P815 mastocytoma cells induced by Artemisia annua pollen, which might provide a new treatment method for allergic diseases.

Allergic sensitization in patients with rhinitis and bronchial asthma in Ningxia region of China.

BACKGROUND: Allergic rhinitis and bronchial asthma are common allergic diseases. The pattern of dominant allergens depends on the degree of urbanization and the geographic region. The present study characterized the allergens of patients with allergic rhinitis and bronchial asthma in Ningxia region of China. METHODS: A total of 309 patients were enrolled in this study. Western blotting assays were performed of the serum samples to evaluate allergen-specific IgE antibody for inhaled and ingested allergens. Statistical analysis was performed to compare the positive rate among different subgroups. RESULTS: Among the 309 patients, 221 of them had positive test results. There were 157 positive cases for ingested allergens and 174 positive cases for inhaled allergens. No significant differences in positive rates were found between the ingested and inhaled allergens. Among the inhaled allergens, Artemisia was the most frequent allergen, followed by fungi and dog hair. Cashew was the most common ingested allergen, followed by crab, mango, and beef. Further analysis showed no significant differences in positive rate between males and females. However, significant differences in positive rate of inhaled and ingested allergens were found between children (1-13 years old) and adults (above 18 years old) (P < .05), while no significant differences were found between the children and teenagers (14-18 years old). For the comparison between teenagers and adults, significant difference in positive rate was found only in the ingested allergen. CONCLUSION: This study provided the characteristics of allergens in Ningxia population, providing clinical and epidemiological data for prevention and treatment of the diseases in the region.