Dexamethasone Inhibits Repair of Human Airway Epithelial Cells Mediated by Glucocorticoid-Induced Leucine Zipper (GILZ)

Background

Glucocorticoids (GCs) are a first-line treatment for asthma for their anti-inflammatory effects, but they also hinder the repair of airway epithelial injury. The anti-inflammatory protein GC-induced leucine zipper (GILZ) is reported to inhibit the activation of the mitogen-activated protein kinase (MAPK)-extracellular-signal-regulated kinase (ERK) signaling pathway, which promotes the repair of airway epithelial cells around the damaged areas. We investigated whether the inhibition of airway epithelial repair imposed by the GC dexamethasone (DEX) is mediated by GILZ.

Methods

We tested the effect of DEX on the expressions of GILZ mRNA and GILZ protein and the MAPK-ERK signaling pathway in human airway epithelial cells, via RT-PCR and Western blot. We further evaluated the role of GILZ in mediating the effect of DEX on the MAPK-ERK signaling pathway and in airway epithelium repair by utilizing small-interfering RNAs, MTT, CFSE labeling, wound-healing and cell migration assays.

Results

DEX increased GILZ mRNA and GILZ protein levels in a human airway epithelial cell line. Furthermore, DEX inhibited the phosphorylation of Raf-1, Mek1/2, Erk1/2 (components of the MAPK-ERK signaling pathway), proliferation and migration. However, the inhibitory effect of DEX was mitigated in cells when the GILZ gene was silenced.

Conclusions

The inhibition of epithelial injury repair by DEX is mediated in part by activation of GILZ, which suppressed activation of the MAPK-ERK signaling pathway, proliferation and migration. Our study implicates the involvement of DEX in this process, and furthers our understanding of the dual role of GCs.     

Online control of cell culture redox potential prevents antibody interchain disulfide bond reduction

The phenomenon of monoclonal antibody (mAb) interchain disulfide bond reduction during manufacturing processes continues to be a focus of the biotechnology industry due to the potential for loss of product, increased complexity of purification processes, and reduced stability of the drug product. We hypothesized that antibody reduction can be mitigated by controlling the cell culture redox potential and subsequently established a threshold redox potential above which the mAb remained intact and below which there were significant and highly variable amounts of reduced mAb. Using this knowledge, we developed three control schemes to prevent mAb reduction in the bioreactor by controlling the cell culture redox potential via an online redox probe. These control methodologies functioned by increasing the concentration of dissolved oxygen (DO), copper (II) (Cu), or both DO and Cu to maintain the redox potential above the threshold value. Using these methods, we were able to demonstrate successful control of antibody reduction. Importantly, the redox control strategies did not significantly impact the cell growth, viability, mAb production, or product quality attributes including aggregates, C-terminal lysine, high mannose, deamidation, and glycation. Our results demonstrate that controlling the cell culture redox potential is a simple and effective method to prevent mAb reduction.     

Molecular character of a phosphatase 2C (PP2C) gene relation to stress tolerance in Arabidopsis thaliana

Protein phosphatases type 2C (PP2Cs) from group A, which includes the ABI1/HAB1 and PP2CA branches, are key negative regulators of ABA signaling. HAI-1 gene had been shown to affect both seed and vegetative responses to ABA, which is one of PP2Cs clade A in Arabidopsis thaliana. Transgenic plants containing pHAI-1::GUS (β-glucuronidase) displayed GUS activity existing in the vascular system of leave veins, stems and petioles. Green fluorescent protein fused HAI-1 (HAI-1-GFP) was found in the nucleus through transient transformation assays with onion epidermal cells. The water-loss assays indicated the loss-of-function mutants did not show symptoms of wilting and they had still turgid green rosette leaves. The assays of seed germination by exogenous ABA and NaCl manifested that the loss-of-function mutants displayed higher insensitivity than wild-type plants. Taken together, the final results suggest that the HAI-1 (AT5G59220) encoded a nuclear protein and it can be highly induced by ABA and wound in Arabidposis, the stress-tolerance phenotype showed a slightly improvement when HAI-1 gene was disrupted.     

灌浆结实期温度对水稻产量和品质形成的影响

灌浆结实期是水稻产量和品质形成的关键时期,该时期温度对水稻籽粒灌浆具有显著的影响.随着全球气候趋暖以及极端天气频发,温度胁迫下籽粒灌浆和稻米品质的响应特征及其生理生化机制是目前稻作研究的热点之一.本文以灌浆结实期温度为切入点,对水稻产量和品质形成的适宜温度与温度影响时段以及温度胁迫下水稻生理生化特征等方面进行了梳理.灌浆初期(齐穗后20 d)是温度影响水稻产量和品质形成的关键时期,适温(21 ～ 26℃)有利于水稻灌浆和淀粉的充实与沉积,过高或过低温度均不利于提高水稻产量和品质.温度胁迫下,水稻生理生化活性下降,光合功能降低,抗逆性减弱,干物质积累和运转受抑,从而造成产量下降及品质变劣.这些可能为水稻优质高产栽培和灌浆结实期温度研究提供一定的参考.     

特有植物多样性分布格局测度方法的新进展

特有植物是生物多样性保护的重要对象,对其分布格局的研究可以为生物多样性优先保护区的确定提供重要参考.研究人员利用多种测度和分析方法,在不同地理区域对特有现象的分布格局开展了大量研究.随着分子系统学方法的不断完善及一些空间统计分析方法的引入,新的生物多样性测度方法应运而生.本文介绍了生物多样性测度方法的类型及其特点、应用现状与前景.这些测度方法的发展经历了从单一的时间或空间格局到时空格局统一的过程,具体涉及物种丰富度、谱系多样性、进化特异性以及这3种测度方法整合空间分布加权的算法.其中,谱系多样性指数(phylogenetic diversity)、谱系特有性指数(phylogenetic endemism)以及空间加权的进化特异性指数(biogeographically weighted evolutionary distinctiveness)尤其值得关注.中国特有植物分布格局的研究需要在以下4个方面进一步开展工作:(1)完善特有物种的分布格局研究;(2)加强物种的测序工作,完善谱系多样性格局的分析;(3)结合系统发育信息,揭示谱系多样性及进化历史的分布格局,进而深入开展物种p多样性和谱系p多样性的研究;(4)加强物种分布区变化的模拟,在时间维度上探讨特有现象的变化格局,为生物多样性保护提供更完善的理论支持.     

体外培养小鼠下颌下腺细胞生长特性的实验研究

目的：研究小鼠下颌下腺细胞的培养方法,探讨下颌下腺细胞培养条件及细胞生长特性,为研究干细胞转分化涎腺腺泡细胞以及涎腺再生研究奠定理论基础和技术支持。方法：取2周龄的小鼠,组织块法和消化法分别进行培养,用活细胞观察法和HE染色法记录细胞形态学特征;免疫荧光染色法鉴定;通过生长曲线和细胞倍增时间来比较两种方法对细胞增殖能力的影响。结果：组织块法和消化法均可以成功的培养下颌下腺细胞,（1）组织块法培养的细胞呈卵圆形或多边形,10天左右成铺路石样;消化法培养细胞亦为上皮样细胞,呈多边形,胞质丰富;（2）HE染色下颌下腺细胞呈多边形,胞核明显;（3）Cytokeratin13和AQP5表达阳性,Wimentin表达阴性;（4）组织块法培养细胞增殖相对较平缓,消化法培养细胞增长较迅速;（5）消化法培养倍增时间（1．3471±0．6071）天比组织块法倍增时间（2．1887±1．1503）明显缩短（P〈0．05）;结论：体外可以成功的培养下颌下腺细胞,但是同时证明下颌下腺细胞长期传代较困难,这为研究干细胞转分化涎腺细胞和涎腺再生体内实验提供了理论和实验基础。