Objective To explore the effects of lipopolysaccharide (LPS) on the expression of inflammatory genes in A549 cells line under different concentrations and different action time, this study laid the foundation for further establishment of acute respiratory distress syndrome (ARDS) cell model in the optimal concentration-time way. Methods A549 cells line was incubated routinely in 5%CO2 incubator at 37 ℃ with high glucose DMEM medium which included 10% fetal calf serum. Cells in logarithmic phase was cultured for passage, the cells was count to adjust cell density to (5-7)×105 and tile evenly in six-hole plate. Cells were cultivated for 2 days and once the cells confluence to 50%-60%, serum-free medium DMEM was changed for 12 hours cultivation. 10 mg LPS was added to 10 mL DMEM for oscillated blending to prepare 1 g/L stock solution. 0.5, 1.0 and 2.5 mL LPS stock solution was taken respectively and diluted LPS stock solution for 50 mL constant volume to prepare 0, 10, 20 and 50 mg/L LPS working solution. Then 0, 10, 20 and 50 mg/L LPS solution was added to react for 0, 1, 3 and 5 hours respectively. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to examine mRNA expression of A549 cells line interleukins (IL-6, IL-1β) and tumor necrosis factor-α(TNF-α). LPS action of 10 mg/L for 0 hour was used as the time control group, LPS action of 0 mg/L for 1 hour was used as the concentration control group, and the gene expression was calculated with 2-ΔΔCt method. Results ① As to the time factor, with the same action of LPS concentration, the relative expression levels of inflammatory genes (IL-6, IL-1β and TNF-α) in A549 cells line after being treated with 10 mg/L LPS for 1 hour were significantly higher than those for 0 hour respectively [IL-6 mRNA (2-ΔΔCt): 5.71±0.42 vs. 1.00±0.00, IL-1β mRNA (2-ΔΔCt): 5.63±0.30 vs. 1.00±0.00, TNF-α mRNA (2-ΔΔCt): 5.38±0.61 vs. 1.00±0.00, all P < 0.01], and there were no significant changes in the expression levels of inflammatory genes in A549 cells line of other time groups. ② As to the concentration factor, with the same action time, the relative expression levels of inflammatory genes (IL-6, IL-1βand TNF-α) in A549 cells line after being treated with 10 mg/L LPS for 1 hour were significantly higher than with 0 mg/L for 1 hour respectively [IL-6 mRNA (2-ΔΔCt): 5.70±0.64 vs. 1.00±0.00, IL-1β mRNA (2-ΔΔCt): 6.25±0.25 vs. 1.00±0.00, TNF-α mRNA (2-ΔΔCt): 5.57±0.25 vs. 1.00±0.00, all P < 0.01], there were no significant changes in the expression levels of inflammatory genes in A549 cells line of other concentration groups. Conclusion The LPS concentration of 10 mg/L and the action time of 1 hour are the most suitable concentration-time conditions for establishing ARDS cell models of A549 cells line.

Objective:To evaluate the nutritional status of patients in intensive care unit (ICU) by using nutritional risk screening 2002 scale (NRS2002), subjective general assessment (SGA) and critical illness nutritional risk score (NUTRIC), and to compare the characteristics and applicability of three scoring tools.Methods:A cross-sectional survey was conducted. 315 patients admitted to the comprehensive ICU of Affiliated Qingdao Municipal Hospital of Qingdao University from April 2018 to July 2019 were enrolled. Basic information of patients was collected, and patients were divided into two groups with 65 years old as the standard to compare the nutritional status of patients among different genders and ages. The nutritional status of patients were assessed by NRS2002, SGA, and NUTRIC. Height, weight, body mass index (BMI), triceps skinfold thickness (TSF), upper arm circumference (AC), leg circumference (LC), and other related parameters of human nutrition were measured. Total protein (TP), albumin (Alb), prealbumin (PA), serum creatinine (SCr), blood urea nitrogen (BUN), total cholesterol (TC), triglyceride (TG), total number of lymphocytes (LYM), hemoglobin (Hb), C-reactive protein (CRP) and other blood biochemical indicators were performed. Spearman rank correlation analysis was used to analyze the correlation between the three nutrition evaluation scales and other objective nutrition parameters. Binary multivariate Logistic regression analysis was used to evaluate the influencing factors of nutritional status with three scales of patients in ICU.Results:Among 315 patients in ICU, 183 were male and 132 were female. There were 143 patients < 65 years old and 172 ≥ 65 years old. In male patients, the acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score, age and BUN of patients aged ≥ 65 years old were significantly increased, and the height, weight, BMI, TSF, AC, LC, Alb and PA were significantly lowered as compared with those aged < 65 years old, while the difference in other indicators was not statistically significant. In the female patients, the APACHEⅡ score, age, SCr and BUN of the patients aged ≥ 65 years old were significantly increased, the height, Alb, PA and Hb were significantly decreased as compared with those aged < 65 years old, and the difference in other indicators was not statistically significant. The proportion of patients with nutritional risk evaluated by NRS2002 (NRS2002 score ≥ 3) was 87.62% (276/315). SGA showed that the proportion of malnourished patients (SGA was grade B or C) was 62.86% (198/315). NUTRIC showed 66.03% of patients (208/315) in high nutritional risk (NUTRIC score ≥ 5). Spearman rank correlation analysis showed that there were significant correlations among NRS2002, SGA and NUTRIC of patients in ICU ( rNRS2002 with SGA = 0.522, rNRS2002 with NUTRIC = 0.392, rSGA with NUTRIC = 0.442, all P < 0.01). Among the three assessment tools, SGA had the best correlation with blood biochemical indicators and body measurements to assess nutritional status, followed by NRS2002, and NUTRIC had the worst correlation. Binary multivariate Logistic regression showed that APACHEⅡ score, BMI, AC, BUN and TG were factors influencing NRS2002 assessment of nutritional status in ICU patients [odds ratio ( OR) were 2.535, 0.404, 1.438, 0.858, and 2.391, respectively, all P < 0.05]; APACHEⅡ score, age, weight, TP, BUN, LYM and CRP were influence factors of SGA for evaluating the malnutrition of patients in ICU ( OR values were 1.074, 1.038, 0.921, 0.947, 1.077, 1.625 and 0.991, respectively, all P < 0.05); APACHEⅡ score, age, LYM and CRP were the influence factors of NUTRIC assessment for malnutrition of patients in ICU ( OR values were 1.159, 1.049, 0.715 and 0.995, respectively, all P < 0.05). Conclusions:The nutrition status of ICU patients evaluated by NRS2002, SGA and NUTRIC was simple and easy to operate, and the positive screening rate of NRS2002 was the highest, which was suitable for patients with mild conditions in ICU. SGA is the most valuable tool to evaluate the nutritional status of ICU patients. NUTRIC has a poor correlation with objective indicators reflecting nutritional status, while its indicators are objective and easy to obtain, which is suitable for ICU patients with critical condition and unclear consciousness. Nutritional assessment tools should be integrated with the patient's gender, age, anthropometric and biochemical indicators.