ABSTRACT
Objective To investigate the effect of hydrochloric acid (HCl) stimulation and mechanical stretch on epithelial-mesenchymal transition (EMT) and hyaluronan (HA) production in human lung epithelial cells. Methods Human lung epithelial cell line BEAS-2B was cultured in vitro, which was divided into phosphate-buffer saline (PBS) + static group, HCl + static group, PBS + stretch group, and HCl + stretch group respectively in the logarithmic phase. The BEAS-2B cells in two stretching groups were challenged by cyclic stretch with 20% amplitude, frequency of 0.33 Hz, sine wave of the FX-5000T system for 48 hours. The morphology changes in cells before and after stretch were observed with inverted microscope. The protein expressions of epithelial markers E-cadherin and cytokeratin-8 (CK-8) as well as mesenchymal markers vimentin and α-smooth muscle actin (α-SMA) were determined by Western Blot. The secretion of HA was determined by enzyme linked immunosorbent assay (ELISA). Results ① It was shown by microscopic observation that BEAS-2B cells displayed cobblestone morphology, linked closely and cell polarity in PBS + static group, which did not change obviously after HCl stimulation alone. Given purely mechanical stretch after 48 hours, the cells morphology changed from cobblestone shape into long spindle, and increased intercellular space obviously. Double hit of HCl and stretch changed the cells morphology more significantly. ② It was shown by Western Blot that compared with the PBS + static group, HCl alone or combined with purely mechanical stretch after 48 hours, the expressions of E-cadherin and CK-8 were decreased, while those of vimentin and α-SMA were increased, and it was more pronounced in HCl + stretch group [the expression quantity (gray value) as base 1 in PBS + static group, E-cadherin: 0.16±0.08 vs. 1, CK-8: 0.10±0.03 vs. 1, vimentin: 3.35±0.38 vs. 1, α-SMA: 3.10±0.45 vs. 1, all P < 0.01]. ③ It was shown by ELISA that both HCl stimulation and stretch could induce BEAS-2B cells secreting HA as compared with PBS + static group (μg/L: 55.763±0.687, 63.005±0.493 vs. 49.876±1.867), and the production of HA increased more remarkably after double hit (μg/L: 78.220±1.085 vs. 49.876±1.867, P < 0.01). Conclusions Both HCl and mechanical stretch could induce EMT and increase HA secretion in human lung epithelial cells in vitro. Double hit of HCl stimulation and mechanical stretch induced EMT apparently, and further increased the production of HA.
ABSTRACT
Objective To analyze the potential factors affecting the prognosis of patients with sepsis and evaluate their values in predicting the disease outcome. Methods A clinical prospective study was conducted. Fifty-three septic patients admitted to intensive care unit (ICU) of the First Affiliated Hospital of Guangzhou Medical University from October 17th, 2012 to August 8th, 2013 were enrolled, and in the same term 35 volunteers having passedphysical check-up were assigned in the healthy control group. According to the severity of the patients, they were divided into sepsis, severe sepsis and septic shock groups. Furthermore, based on the difference in scores of acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ), the patients were divided into low-risk (APACHE Ⅱ scores 0.05). The IL-8 level of non-coagulation defect group was significantly lower than that of adjusted (ng/L:24.67 vs. 27.23, P0.05). Conclusions The grade of sepsis severity, APACHEⅡscore, whether existence of coagulation dysfunction being present or not and whether its presence being adjusted or not during the septic patients' stay in ICU, the levels of blood lactate, PCT, IL-6 and IL-8 on the first day in ICU are significantly correlated to the prognosis of septic patients. Whether the existence of coagulation dysfunction being present or not, whether coagulation dysfunction being adjusted or not and the blood lactate level are the independent prognostic factors of septic patients, and the plasma concentrations of IL-6 and IL-8 are the independent affecting factors of whether coagulation dysfunction occurring or not, therefore they have predicting value concerning the occurrence of coagulation dysfunction in septic patients.
ABSTRACT
Objective To explore the causes of rising carbapenem resistance in Acinetobacter baumannii isolates by analysis of the carbapenemase genotypes in A .baumannii .Methods WHONET 5 .4 was used to analyze the changing resistance profile of A .baumannii isolates over years .A total of 320 carbapenem‐resistant A .baumannii isolates were collected from the patients in an intensive care unit from January 2008 to December 2012 .All strains were identified and tested by VITEK 2 for their susceptibility .The metallo‐β‐lactamases genes and OXA carbapenemase genes were investigated by polymerase chain reaction . Results The resistance rate of A .baumannii isolates to meropenem and imipenem increased rapidly from 10 .8% to 80 .4% and from 13 .5% to 83 .5% during the period from 2008 and 2012 .The prevalence of blaOXA‐23and ISAba1‐associated blaOXA‐23genes increased from 25 .0% to 97 .1% .No metallo‐β‐lactamase encoding genes were detected .Conclusions Our results indicate that the rising carbapenem resistance in the A . baumannii isolates in our intensive care unit may be associated with the high prevalence of blaOXA‐23 and IS A ba1‐associated blaOXA‐23 genes .