Caffeine Ameliorates Age-Related Hearing Loss by Downregulating the Inflammatory Pathway in Mice.

OBJECTIVE: Age-related hearing loss (ARHL), also known as presbycusis, is a debilitating sensory impairment that affects the elderly population. There is currently no ideal treatment for ARHL. Long-term caffeine intake was reported to have anti-aging effects in many diseases. This study is to identify whether caffeine could ameliorate ARHL in mice and analyze its mechanism. METHODS: Caffeine was administered in drinking water to C57BL/6J mice from the age of 3 months to 12 months. The body weight, food intake and water intake of the mice were monitored during the experiment. The metabolic indicators of serum were detected by ELISA. The function of the hearing system was evaluated by ABR and hematoxylin and eosin staining of the cochlea. Genes' expression were detected by Q-PCR, immunofluorescencee and Western blot. RESULTS: The results showed that the ARHL mice exhibited impaired hearing and cochlear tissue compared with the young mice. However, the caffeine-treated ARHL mice showed improved hearing and cochlear tissue morphology. The expression of inflammation-related genes, such as TLR4, Myd88, NF-κB, and IL-1ß, was significantly increased in the cochleae of ARHL mice compared with young mice but was down-regulated in the caffeine-treated cochleae. CONCLUSIONS: Inflammation is involved in ARHL of mice, and long-term caffeine supplementation could ameliorate ARHL through the down-regulation of the TLR4/NF-κB inflammation pathway. Our findings provide a new idea for preventing ARHL and suggest new drug targets for ARHL treatment.

Leucobacter chinensis sp. nov., with plant growth-promoting potential isolated from field soil after seven-years continuous maize cropping.

A novel Gram-stain-positive, aerobic, non-motile and rod-shaped bacterium, designated strain NC76-1T, was isolated from soil from a field that had undergone seven years continuous maize cropping from Liuba town located in Zhangye city, Gansu province, PR China. Colonies of strain NC76-1T were white, opaque and circular with a convex shape. The isolate was found to be able to grow at 10-40 °C (optimum 30 °C), pH 6.0 to 12.0 (optimum 7.0-8.0) and with 0-5.0 % (w/v) NaCl (optimum 0%). On the basis of the results of 16S rRNA gene sequence analysis, the strain fell within the clade of the genus Leucobacter, showing the highest sequence similarities with Leucobacter iarius 40T (97.4%), Leucobacter aridicollis CIP 108388T (97.0%), Leucobacter chromiireducens subsp. solipictus TAN 31504T (96.7%) and Leucobacter denitrificans M1T8B10T (96.7%). The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between NC76-1T and its closest relatives, L. iarius 40T, L. aridicollis CIP 108388T, L. chromiireducens subsp. solipictus TAN 31504T and L. denitrificans M1T8B10T were ≤73.5 % and 20.3%, respectively. The genomic DNA G+C content of NC76-1T was 61.5 mol%. It presented MK-11 as the predominant menaquinone. The major cellular fatty acids were anteiso-C15 : 0 (49.2 %) and iso-C16 : 0 (35.7%). The major polar lipids were found to be diphosphatidyglycerol, phosphatidylglycerol, phosphatidylethanolamine, aminoglycolipid, five glycolipid and one unidentified lipids. The cell wall amino acids were 2,4-diaminobutyric acid, alanine, glutamic acid, glycine and threonine. On the basis of the phylogenetic, phenotypic and chemotaxonomic characteristics, strain NC76-1T is concluded to represent a novel species within the genus Leucobacter, for which the name Leucobacter chinensis sp. nov. is proposed. The type strain is NC76-1T (GDMCC 1.2286T= JCM 34651T).

Microbacterium sulfonylureivorans sp. nov., isolated from sulfonylurea herbicides degrading consortium.

A novel Gram-stain positive, aerobic, motile, rod-shaped bacterium, designated strain LAM7116T was isolated from a sulfonylurea herbicides degrading consortium enriched with birch forest soil from Xinjiang. Phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain LAM7116T was closely related to the members of the genus Microbacterium, with the highest similarity to Microbacterium flavescens DSM 20643T (98.48%) and Microbacterium kitamiense Kitami C2T (98.48%). Strain LAM7116T formed a distinct subclade with M. flavescens DSM 20643T within the genus Microbacterium in the 16S rRNA gene phylogenetic trees. The genomic DNA G + C content of LAM7116T was 69.9 mol%. The digital DNA-DNA hybridization (dDDH) value between strain LAM7116T and M. flavescens DSM 20643T was 27.20%. The average nucleotide identity (ANI) value was 83.96% by comparing the draft genome sequences of strain LAM7116T and M. flavescens DSM 20643T. The major fatty acids were anteiso-C15:0, anteiso-C17:0, iso-C17:0, and iso-C16:0. The respiratory menaquinones of strain LAM7116T were MK-13 and MK-14. The main polar lipids were diphosphatidylglycerol, phosphatidylglycerol, an unidentified lipid, and an unidentified glycolipid. The peptidoglycan contained the amino acids glycine, lysine, alanine, and glutamic acid. Based on the phenotypic characteristics and genotypic analyses, we consider that strain LAM7116T represents a novel species, for which the name Microbacterium sulfonylureivorans sp. nov. was proposed. The type strain is LAM7116T (= CGMCC 1.16620T = JCM 32823T). Strain LAM7116T secreted auxin IAA and grew well in Ashby nitrogen-free culture medium. Genomic results showed that strain LAM7116T carried the nitrogenase iron protein (nifU and nifR3) gene, which indicated that strain LAM7116T has the potential to fix nitrogen and promote plant growth. At same time, strain LAM7116T can degrade nicosulfuron (a kind of sulfonylurea herbicides) using glucose as carbon source. Microbacterium sulfonylureivorans sp. nov. LAM7116T is a potential candidate for the biofertilizers of organic agriculture areas, and may possess potential to be used in bioremediation of nicosulfuron-contaminated environments.

Response of bacterial communities in saline-alkali soil to different pesticide stresses.

The objective is to understand the diversity of bacteria-degrading pesticide pollutants in Xinjiang saline-alkali soil environment and resolve the lack of suitable degrading bacteria resources for bioremediation of pesticide pollution in this environment. The soil of long-term continuous cropping cotton fields in Xinjiang was used to culture the degrading bacterial communities under long-term stress of five pesticides, such as beta-cypermethrin. Then, the degradation rate and structural composition of each bacterial communities were analyzed. The soil bacterial diversity in Xinjiang saline-alkali cotton fields was high, from which not only imidacloprid and other commonly and once used pesticide-degrading bacterial communities were enriched but also isoprocarb-degrading bacterial communities, which had never been used, were enriched. After long-term passage, the structural composition of each degrading bacterial communities was stable, and the degradation rates were between 17 and 48%, respectively, in a specific culture period. Each degrading bacterial communities covers many reported pesticide-degrading bacterial genera and contains unique bacterial genera in each 3. These results laid a foundation for studying the metabolic pathway of pesticide pollutants in saline-alkali environment and exploring microbial resources in Xinjiang. Graphical Abstract Variety of pesticide degrading bacteria resources in saline alkali soil of Xinjiang.

Pseudomonas nicosulfuronedens sp. nov., a nicosulfuron degrading bacterium, isolated from a microbial consortium.

A Gram-stain-negative, aerobic, motile, short-rod-shaped bacterium with nicosulfuron-degrading ability, designated strain LAM1902T, was isolated from a microbial consortium enriched with nicosulfuron as a sole nitrogen and energy source. The optimal temperature and pH for growth of strain LAM1902T were 30 °C and pH 6.0, respectively. Strain LAM1902T could grow in the presence of NaCl with concentration up to 4.0 % (w/v). Comparative analysis of 16S rRNA gene sequences revealed that LAM1902T was closely related to the members of the family Pseudomonadaceae to the genus Pseudomonas, with the highest similarity to Pseudomonas nitroreducens DSM 14399T (99.6 %), Pseudomonas nitritireducens WZBFD3-5A2T (99.3 %) and Pseudomonas panipatensis Esp-1T (98.8 %). Multi-locus sequence analysis based on both concatenated sequences of the 16S rRNA gene and three housekeeping genes (gyrB, rpoB and rpoD) further confirmed the intrageneric phylogenetic position of strain LAM1902T. The genomic DNA G+C content of LAM1902T was 64.8 mol%. The low values of in silico DNA-DNA hybridization (less than 43.7 %) and average nucleotide identity (less than 90.9 %) also showed that the strain was distinctly different from known species of the genus Pseudomonas. The major fatty acids were C16 : 0, C17 : 0 cyclo and anteiso C15 : 0. Ubiquinone Q-9 was detected as the predorminant respiratory quinone. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and aminophospholipid. Based on phylogenetic, phenotypic and chemotaxonomic analyses and genome comparisons, we conclude that strain LAM1902T represents a novel species, for which the name Pseudomonas nicosulfuronedens sp. nov. is proposed. The type strain is LAM1902T (=JCM 33860T=KCTC 72830T).

Role of Endoplasmic Reticulum Stress in Otitis Media.

Endoplasmic reticulum (ER) stress occurs in many inflammatory responses. Here, we investigated the role of ER stress and its associated apoptosis in otitis media (OM) to elucidate the mechanisms of OM and the signaling crosstalk between ER stress and other cell damage pathways, including inflammatory cytokines and apoptosis. We examined the expression of inflammatory cytokine- and ER stress-related genes by qRT-PCR, Western blotting, and immunohistochemistry (IHC) in the middle ear of C57BL/6J mice after challenge with peptidoglycan polysaccharide (PGPS), an agent inducing OM. We also evaluated the effect of the suppression of ER stress with tauroursodeoxycholic acid (TUDCA), an ER stress inhibitor. The study revealed the upregulation of ER stress- and apoptosis-related gene expression after the PGPS treatment, specifically ATF6, CHOP, BIP, caspase-12, and caspase-3. TUDCA treatment of PGPS-treated mice decreased OM; reduced the expression of CHOP, BIP, and caspase 3; and significantly decreased the proinflammatory gene expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). These results suggest that PGPS triggers ER stress and downstream proinflammatory gene expression in OM and that inhibition of ER stress alleviates OM. We propose that ER stress plays a critical role in inflammation and cell death, leading to the development of OM and points to ER stress inhibition as a potential therapeutic approach for the prevention of OM.

Comparative efficacy of six therapies for Hypopharyngeal and laryngeal neoplasms: a network meta-analysis.

BACKGROUND: Hypopharyngeal and laryngeal neoplasms are both fatal and hard to catch in early stages. Yet which treatment is the most efficacious one still remain unanswered. This network meta-analysis (NMA) was conducted to investigate effectiveness of six therapies being utilized in clinical practice nowadays. METHODS: PubMed and Embase were retrieved to synthesize data. Direct and indirect evidence was combined to compare efficacy of treatments. A relative ranking of the six regimens was calculated by the surface under the curve ranking area (SUCRA). RESULTS: A total of 28 trials with 9109 patients were included in our NMA. Five endpoints investigated included 3/5-year overall survival (3/5-OS), 3/5-year disease free survival (3/5-DFS) and 5-year overall survival rate (5-OSR). In terms of all efficacy outcomes, radiotherapy combined with surgery (RT + S) proved to be better than other therapies while radiotherapy (RT) alone also performed well. Induction chemoradiotherapy (ICRT) was the best regarding 3-DFS (SUCRA = 0.846) while current chemoradiotherapy (CCRT) ranked first in 5-DFS (SUCRA = 0.933) according to SUCRA results. No significant differences were demonstrated in 5-DFS and 5-OSR as shown in the results of NMA. CONCLUSIONS: RT combined with surgery turned out to be optimal therapy of all the outcomes while the efficacy of RT was relatively poorer in the treatment of patients with larynx stage III-IV and hypopharynx stage II-IV. Also, the good performance of CCRT and ICRT in terms of DFS made them as secondary recommended therapies. There is no significant difference between surgery and transoral laser microsurgery (TLM) alone.

[Correlation of osteopontin expression and laryngeal squamous cell carcinoma infiltration and metastasis].

OBJECTIVE: To investigate osteopontin (OPN) expression in plasma and tissue of patients with layngeal squamous cell carcinoma and analyze its role in invasion, metastasis, and clinical significance in laryngeal quamous cell carcinoma. METHOD: Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry were used to detect expression of OPN in plasma and tissue of 60 cases of laryngeal squamous cell carcinoma, 20 cases of adjacent normal laryngeal tissue and 20 cases of plasma from healthy subjects. RESULT: The expression of plasma OPN was closely correlated with clinical stage and cervical lymphatic metastasis in laryngeal squamous cell carcinoma (P < 0.05), but no significant correlation with the tumor location, pathological grade, gender and age (P > 0.05). The expression of OPN increased in plasma during cancer development: laryngeal squamous cell carcinoma (38.089 ± 9.225) ng/ml, healthy subjects (18.563 ± 9.308) ng/ml. There was a significant difference between the groups (P < 0.05). The expression of OPN in tissue was closely correlated with clinical stage (P < 0.05), pathological grade (P < 0.05) and cervical lymphatic metastasis (P < 0.05) in laryngeal squamous cell carcinoma adjacent atypical hyperplastic epithelium and carcinoma. The expression of OPN increased in tissue during cancer development: laryngeal squamous cell carcinoma (56.67%), adjacent normal laryngeal tissue (15.00%). There was a significant difference between the groups (P < 0.05). Elevated expression of plasma OPN is positively correlated with the expression of OPN in tissue in laryngeal squamous cell carcinoma patients (r = 0. 871, P < 0.05). CONCLUSION: OPN plays an important role in the infiltration, metastasis and carcinogenesis in laryngeal squamous cell carcinoma. Combination of serum OPN, tissue OPN detection can be used as diagnostic and surveillance indicators for laryngeal squamous cell carcinoma infiltration and metastasis.

Correlation of osteopontin expression and laryngeal squamous cell carcinoma infiltration and metastasis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate osteopontin (OPN) expression in plasma and tissue of patients with layngeal squamous cell carcinoma and analyze its role in invasion, metastasis, and clinical significance in laryngeal quamous cell carcinoma.@*METHOD@#Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry were used to detect expression of OPN in plasma and tissue of 60 cases of laryngeal squamous cell carcinoma, 20 cases of adjacent normal laryngeal tissue and 20 cases of plasma from healthy subjects.@*RESULT@#The expression of plasma OPN was closely correlated with clinical stage and cervical lymphatic metastasis in laryngeal squamous cell carcinoma (P 0.05). The expression of OPN increased in plasma during cancer development: laryngeal squamous cell carcinoma (38.089 ± 9.225) ng/ml, healthy subjects (18.563 ± 9.308) ng/ml. There was a significant difference between the groups (P < 0.05). The expression of OPN in tissue was closely correlated with clinical stage (P < 0.05), pathological grade (P < 0.05) and cervical lymphatic metastasis (P < 0.05) in laryngeal squamous cell carcinoma adjacent atypical hyperplastic epithelium and carcinoma. The expression of OPN increased in tissue during cancer development: laryngeal squamous cell carcinoma (56.67%), adjacent normal laryngeal tissue (15.00%). There was a significant difference between the groups (P < 0.05). Elevated expression of plasma OPN is positively correlated with the expression of OPN in tissue in laryngeal squamous cell carcinoma patients (r = 0. 871, P < 0.05).@*CONCLUSION@#OPN plays an important role in the infiltration, metastasis and carcinogenesis in laryngeal squamous cell carcinoma. Combination of serum OPN, tissue OPN detection can be used as diagnostic and surveillance indicators for laryngeal squamous cell carcinoma infiltration and metastasis.

[Prognostic analysis of 450 patients with lung squamous cancer].

OBJECTIVE: To study the treatment strategy and prognosis and its affected factors of lung squamous cancer according retrospective analysis. METHODS: Clinic data of 450 lung squamous cancer inpatient cases who were performed complete resection from January 2004 to January 2007, was retrospectively reviewed. There were 363 male and 87 female patients, aged from 31 to 82 years, with a mean of 60.5 years and a median of 62 years. RESULTS: The overall 5-year survival rate was 52.4%. Cox Regression suggested that preoperative N status (χ(2) = 18.969, P = 0.000), N stage (χ(2) = 44.069, P = 0.000) and TNM stage (χ(2) = 63.025, P = 0.000) are independent factors affecting the prognosis. Adjuvant chemotherapy affects the prognosis of stage II-IIIA lung squamous cancer (5-year survival rate: 48.9% vs. 37.7%, χ(2) = 3.946, P = 0.047). Studying the combined therapy of stage IIIA, the chemoradiotherapy group achieved the best survival (48.8%), then single chemotherapy group (35.9%) and no treatment group (28.5%), and the single radiotherapy group achieved the poorest survival rate (11.1%), and there were statistically significant differences among them (χ(2) = 8.397, P = 0.038). CONCLUSIONS: The 5-year survival rate of lung squamous cancer has significantly increased through promoting the standard of operation, especially increasing the standard of lymph node dissection. Adjuvant chemotherapy is benefit for stage II-IIIA patients and combined chemoradiotherapy is the best choice for stage IIIA patients. If preoperative examination suggests mediastinal lymph node's enlargement and fusion, the operation should not be performed.

[Relationship and expression of hepatocyte growth factor and vascular endothelial growth factor in serum preoperative and postoperative patients with laryngeal squamous cell cancer].

OBJECTIVE: To investigate the role and clinical value of the serum level of hepatocyte growth factor (HGF) and vascular endothelial growth factor(VEGF) in laryngeal squamous cell cancer and the relationship between HGF, VEGF and clinicopathological factors. METHOD: We measured serum HGF and VEGF level in 54 laryngeal squamous cell cancer patients using ELISA to demonstrate the variation of HGF, VEGF level before operation and at day 1, 3, 7, 14 post-surgery. The concentration of the serum HGF, VEGF content in 35 normal healthy people and in 30 vocal cords polyps patients were measured by the quantitative sandwich enzyme linked immunoassay technique. RESULT: (1) The level of serum HGF and VEGF in the patients with laryngeal squamous cell cancer was significantly higher than that of normal healthy people and vocal cord polyps patients (P < 0.01). (2) The level of serum HGF, VEGF in the patients with laryngeal squamous cell cancer was related to clinical stage, differentiation situation, lymph nodes metastasis (P < 0.01), but it was independent of age at diagnosis, gender and tumor location (P > 0.05). (3) There was a significantly positive correlation between preoperative serum VEGF and HGF levels (r = 0.7667, P < 0.01). (4) Post-operational serum HGF levels in 54 laryngeal squamous cell cancer patients who underwent surgical intervention increased significantly, peaked at day 3 after operation. Serum HGF levels of survivors during follow-up period gradually decreased at day 7 and day 14 after operation. The postoperative serum levels of VEGF were decreased significantly than that of preoperation. CONCLUSION: HGF and VEGF may play an important role in the development and progression of human laryngeal cancer. Elevated serum HGF and VEGF levels predict a more aggressive biological behavior in laryngeal squamous cell cancer.

Endomorphins restored the endothelium-dependent relaxation of the rabbit aorta rings exposed to high D-glucose condition via NO-cGMP pathway.

Rings of rabbit aorta that were both incubated in a high concentration of D-glucose and contracted submaximally by phenylephrine showed significantly decreased endothelium-dependent relaxations induced by acetylcholine. The cGMP production of aorta rings was also reduced. Treatment with endomorphins (1-1000 nmol/L) restored acetylcholine-induced relaxations of aorta rings incubated in high glucose concentrations and increased the cGMP synthesis. Moreover, this effect of endomorphins on endothelium was antagonized by naloxone, and the increase in the production of cGMP was also blocked.