Knockdown of phosphoinositide-dependent kinase 1 (PDK1) inhibits fibrosis and inflammation in lipopolysaccharide-induced acute lung injury rat model by attenuating NF-κB/p65 pathway activation.

BACKGROUND: Acute lung injury (ALI) is a common inflammatory disease of the lung. This study aimed to investigate the effect of 3-phosphoinositide-dependent kinase 1 (PDK1) interference on the levels of fibrosis and proinflammatory factors in lipopolysaccharide (LPS)-induced ALI and discuss the relevant mechanism. METHODS: An ALI model was established by intravenous injection of LPS treatment. A total of 24 Sprague-Dawley (SD) rats were randomly divided into 4 groups: sham group; ALI group; ALI + shRNA-NC group; and ALI + PDK1-shRNA group. Lung injury score, minute ventilation, lung volume, and airway resistance were used to evaluate lung function injury. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect PDK1 messenger RNA (mRNA) level. Western blot was performed to detect expression levels of PDK1, transforming growth factor-ß (TGF-ß), α-smooth muscle actin (α-SMA), toll-like receptor 4 (TLR4), p65, and myeloid differentiation primary response gene 88 (MyD88). The contents of interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), and monocyte chemoattractant protein-1 (MCP-1) were detected by enzyme-linked immunosorbent assay (ELISA). The pathological changes and fibrosis of lung tissues were estimated by hematoxylin and eosin (H&E) and Masson staining. RESULTS: The results revealed that high lung injury score, low minute ventilation, low lung volume, and small airway resistance were present in the ALI group. Likewise, severe histopathological damage and fibrosis were apparent in the ALI group. Otherwise, contents of TNF-α, iNOS, IL-6, MCP-1, and levels of α-SMA, TGF-ß, TLR4, phosphorylated (p)-p65, and MyD88 were enhanced in the ALI group. Interestingly, pathological changes and fibrosis were improved significantly in the ALI + PDK1-shRNA group. Besides, knockdown of PDK1 reduced lung injury score and enhanced minute ventilation, lung volume, and airway resistance. Moreover, knockdown of PDK1 decreased the contents of TNF-α, iNOS, IL-6, MCP-1, and levels of TGF-ß, α-SMA, TLR4, p-p65, and MyD88. CONCLUSIONS: Knockdown of PDK1 protects LPS-induced ALI via attenuating activation of the nuclear factor-κB (NF-κB)/p65 pathway.

Dynamic measurement of amnion thickness during loading by speckle pattern interferometry.

INTRODUCTION: In previous studies on the mechanical parameters of amnions (AM), there is a limitation due to the lack of an accurate thickness measurement, which is an important parameter for determining AM-specific mechanical properties. As a bottleneck, the characterization of the basic mechanical properties of AM are greatly restricted, even with the proposal of fracture criteria. METHOD: First, the initial thickness of the AM is estimated by the interpolated-volume-area method. Second, through combinations of our self-developed mini-biaxial tensile device with speckle pattern interferometry, this is the first time that researchers can accurately obtain the AM thickness at each transient moment in the process of loading. RESULTS: Based on the experimental results, an accurate stress-strain curve could be obtained. Two important mechanical parameters-the fracture energy density and amnion rupture modulus-could be extracted as 0.184±0.036MPa and 108.57±17.32MPa, respectively. The fracture energy density and amnion rupture modulus provide objective criteria and a scientific basis for the evaluation of AM rupture. DISCUSSION: The tensile stress-strain curve of a normal human amnion shows a distinct J-shape. This proves that the experimental results are basically reliable. Both important parameters --the fracture energy density and amnion rupture modulus, can be calculated from the stress-strain curve. Extracting these two parameters is critical for the evaluation and prediction of ROM, PROM and PPROM.

How the Crosslinking Agent Influences the Thermal Stability of RTV Phenyl Silicone Rubber.

In this work, a thermal degradation mechanism of room temperature vulcanized (RTV) phenyl silicone rubber that was vulcanized by different crosslinking agents was discussed. Firstly, RTV phenyl silicone rubber samples were prepared by curing hydroxyl-terminated polymethyldiphenylsiloxane via three crosslinking agents, namely, tetraethoxysilane (TEOS), tetrapropoxysilane (TPOS), and polysilazane. Secondly, the ablation properties of RTV phenyl silicone rubber were studied by the muffle roaster test and FT-IR. Thirdly, thermal stability of the three samples was studied by thermogravimetric (TG) analysis. Finally, to explore the thermal degradation mechanism, the RTV phenyl silicone rubber vulcanized by different crosslinking agents were characterized by TG analysis-mass spectrum (TG-MS) and pyrolysis gas chromatogram-mass spectrum (pyGC-MS). Results showed that the thermal stability of RTV phenyl silicone rubber is related to the amount of residual Si⁻OH groups. The residual Si⁻OH groups initiated the polysiloxane chain degradation via an 'unzipping' mechanism.