支气管哮喘患儿lncRNA PVT1和炎症因子表达与激素治疗敏感性的关系研究

目的分析长链非编码RNA中浆细胞瘤多样性异位基因1(lncRNA PVT1)、炎症因子与支气管哮喘患儿激素治疗敏感性的关系。方法选取2019年1月-2019年12月张家口市第一医院收治的60例支气管哮喘患儿作为研究对象,根据不同的激素治疗敏感性分为敏感组和抵抗组,各30例。采集两组患儿气管平滑肌细胞进行原代培养,使用RT-PCR检测lncRNA PVT1相对表达量,ELISA法检测血清炎症因子及siRNA沉默lncRNA PVT1前后原代培养细胞的炎症因子表达水平。结果抵抗组血清及原代培养细胞lncRNA PVT1较敏感组高(P<0.05)。抵抗组血清IL-1β、IL-6、IL-10及TNF-α表达水平较敏感组高(P<0.05)。抵抗组沉默前后原代培养细胞中IL-1β、IL-6、IL-10及TNF-α表达水平的差值较敏感组高(P<0.05)。lncRNA PVT1[OlR=0.145(95% CI:0.057,0.762),P=0.029]、IL-10[OlR=2.853(95% CI:1.542,5.969),P=0.017]及TNF-α[OlR=4.482(95% CI:1.404,13.858),P=0.031]为支气管哮喘患儿激素治疗敏感性的影响因素(P<0.05)。血清lncRNA PVT1、IL-10与TNF-α联合预测支气管哮喘患儿发生激素治疗抵抗的敏感性为91.25%(95% CI:0.835,0.986),特异性为88.74%(95% CI:0.810,0.965),AUC为0.955(95% CI:0.903,1.000),且较单独指标预测的AUC高(P<0.05)。结论过表达lncRNA PVT1可明显加剧支气管哮喘患儿的气道炎症反应,进而降低气管平滑肌对激素治疗的敏感性。

Expression ofα2-Na/K-ATPase in C57BL/6J Mice Inner Ear and Its Relationship with Age-related Hearing Loss

K^(+)cycling in the cochlea is critical to maintain hearing.Many sodium-potassium pumps are proved to participate in K^(+)cycling,such as Na/K-ATPase.Theα2-Na/K-ATPase is an important isoform of Na/K-ATPase.The expression ofα2-Na/K-ATPase in the cochlea is not clear.In this study,we used C57BL/6 mice as a model of presbycusis and implemented immunohistochemistry staining and quantitative real time-PCR,and theα2-Na/K-ATPase expression pattern was confirmed in the inner ear.It was foundα2-Na/K-ATPase was expressed widely in cochlea and its mRNA and protein expression was gradually reduced with aging(4-,14-,26-and 48-weeks old mice).We suspected that,the down-regulation ofα2-Na/K-ATPase expression might be associated with the remodeling of K^(+)cycling,degeneration of morphological structure and decrease of hearing function in aging C57 mice.In conclusion,we speculated that the reduction ofα2-Na/K-ATPase might play an important role in the pathogenesis of age-related hearing loss.

Expression Patterns and Implications of LaminBl in Rat Cochleae

LaminB1,a major component of the nuclear lamina,is a potent regulator of cellular proliferation and senescence and also known to be essential for neuronal migration and brain development.However,the expression patterns of LaminB1 in the rat cochleae are still not fully revealed.Utilizing immunofluorescence.Western blotting,and quantitative real-time PCR,we identified the distribution and expression of LaminB1 in the rat cochleae.Immunofluorescence staining indicated that LaminBl was mainly localized in the auditory hair cells(HCs),spiral ganglion cells(SGC),stria vascularis(STV,including spiral ligament),Reissner's membrane(RM),and limbus laminae spiralis(LLS).Western blotting analysis illustrated that the distribution of LaminBl in rat cochleae was characterized by tissue specificity.The LaminB1 protein was expressed more in SGC and basilar membrane(BM)than in STV.Meanwhile,the mRNA expression of LaminB1 displayed difference in cochlear tissues.These observations preliminarily revealed the expression patterns of LaminB1,providing a theoretical basis for further study on the role of LaminB1 in auditory function.

Interference of Quinolinate Phosphoribosyltransferase Gene QPT Affects Agronomic Traits and Leaf Quality in Nicotiana tabacum L.

Quinolinate phosphoribosyltransferase(QPRTase),a key enzyme in ensuring nicotinic acid is available for the synthesis of defensive pyridine alkaloids in Nicotiana species,also plays an important role in nicotinamide adenine dinucleotide(NAD)biosynthesis.In this study,the morphological traits,the quality characteristics and photosynthetic parameters in QPT-overexpressing/interfering tobacco plants were investigated,respectively.Results showed that the interference of QPT gene not only reduced significantly the morphological traits including plant height,stem girth,leaf number and leaf length,etc.at 20 days after transplanting(DAT),but the flowering period was delayed 10-15 d in interfered tobaccos compared with the overexpressed,control and wild-type counterparts.However,at 40 DAT and 60 DAT,only three indexes(plant height,stem girth and leaf number)in QPT-interfering plants appeared significant difference in comparison with other three types of tobacco lines.Meanwhile,the determination results from nicotine,sugar,K+and Cl-content showed the nicotine content in interfered plants was always significantly lower than that in overexpressed plants,control and the wild-type ones respectively whatever toppling or not.At the same time,the toppling treatment also caused the increasement of K+content among the four different tobacco lines,but the maximum increase amplitude of K+content was found in QPT-overexpressing tobaccos while the minimum appeared in QPT-interfering plants.Finally,QPT-interference in transgenic tobaccos likewise affected the photosynthesis by reducing net photosynthetic rate(Pn),stomata conductance(Gs)and transpiration rate(Tr),while there was no significant difference between QPT-overexpressing plants and the controls and the wild-types.

Downregulation of Cav3.1 T-type Calcium Channel Expression in Age-related Hearing Loss Model

Objective Age-related hearing loss(AHL),characterized by degeneration of cochlea structures,is the most common sensory disorder among the elderly worldwide.The calcium channel is considered to contribute to normal hearing.However,the role of the T-type voltage-activated calcium channel,Cav3.1,remains unclear in AHL.Here,we investigate the age-related change of Cav3.1 expression in the cochlea and D-gal-induced senescent HEI-OC1 cells.Methods Cochleae from C57BL/6 mice at 2 months and 12 months of age were assessed.Senescence in House Ear Institute-Organ of Corti 1(HEI-OC1)cells was induced by D-gal treatment.The immunofluorescence technique was employed to investigate the distribution of Cav3.1 in vivo and in vitro.Quantitative assessment was achieved by Western blotting and real-time PCR.Results In comparison with 2-month-old animals,12-month old C57BL/6 mice exhibited great loss of hair cells and elevated auditory brainstem threshold.The Cav3.1 was located in hair cells,spiral ganglion cells,lateral walls,and the expression of Cav3.1 protein and mRNA decreased in the aged cochleae.D-gal-induced senescence assay confirmed the down-regulation of Cav3.1 expression in senescent HEI-OC1 cells.Conclusion Our results show that age-related down-regulated expression of Cav3.1 in the cochleae is associated with AHL and may contribute to the pathogenesis of AHL.