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Objective To study the relationship between heparin-binding protein(HBP) and β2 integrin on the surface of neutrophil in acute lung injury.Methods A total of 30 mice from animal center of Tongji University were divided into control group,CLP group and antibody-treatment group by random number method.Acute lung injury animal model was established by cecal Ligation and puncture in the mice of CLP group and antibody-treatment group.Sham operations were performed on the mice in control group.Mice in antibody-treatment group received anti-CD18 antibody injected via tail vein 30 min before establishing acute lung injury animal model.Twenty-four hours after operation,mice were sacrificed and lung tissue was taken.After HE staining,lung injury were evaluated by Smith score.Lung tissue wet/dry weight ratio,BALF protein level,plasma HBP level and β2 integrin on neutrophil were measured.T test were used to compare the difference among groups.Pearson correlation was used to study the correlation between HBP,integrin and lung injury index.Results Compared with control group,mice in CLP group had higher Smith score(t=10.607,P＜0.01),lung wet/dry ratio(t=3.968,P=0.001),BALF protein level(t=4.331,P＜0.01) and as well as higher plasma HBP(t=3.515,P=0.002) and β2 integrin(t=4.816,P＜0.01) level.After CD18 antibody treatment,anti-treat group mice had lower Smith score (t=2.307,P=0.033),lung wet/dry ratio(t=3.080,P=0.006),BALF protein level(t=2.484,P=0.023) and as well as higher plasma HBP level(t=2.218,P=0.046) than mice in CLP group.Pearson correlation analysis showed HBP had obvious correlation with wet/dry ratio(r=0.527,P=0.017),BALF protein(r=0.508,P=0.022) and as well as β2 integrin(r=0.674,P=0.001).Conclusions Both HBP and β2 integrin involved in the lung injury pathogenesis.HBP level is interrelated with the severity of lung injury.The β2 integrin is associated with the release of HBP by neutrophil.Inhabiting the function of integrin could decrease HBP level and alleviate the severity of lung injury.
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Objective To investigate the changes in serum level of microparticles (EMPs) in mice with ventilator-induced lung injury (VILI), and explore its significance in VILI. Methods Forty-eight grade SPF male C57BL/6J mice were randomly divided into two groups, with 24 mice in each group: the mice in mechanical ventilation (MV) group were given high tidal volume (VT 30 mL/kg) MV for 4 hours after tracheal intubation, and those in spontaneous breathing group were spontaneously breathed for 4 hours. The apical blood of 12 mice in each group were collected, and serum levels of interleukins (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA), and serum EMPs levels were determined by flow cytometer. The correlations between EMPs and IL-1β, IL-6, and TNF-α were analyzed by linear regression analysis. The lung tissues of other 12 mice in each group were harvested, and wet/dry weight (W/D) ratio was assessed. After hematoxylin-eosin (HE) staining, the morphological changes in lung tissue were observed under light microscope. After double staining of uranium acetate and lead citrate, the ultrastructural changes in lung tissue were observed with electron microscope. Results Compared with spontaneous breathing group, the levels of lung W/D ratio in MV group was significantly increased (5.47±0.14 vs. 4.34±0.11), the levels of IL-1β, IL-6, TNF-α and EMPs were also significantly increased [IL-1β (ng/L): 42.4±4.4 vs. 7.7±3.6, IL-6 (ng/L): 1 239.5±66.3 vs. 21.7±4.6, TNF-α (ng/L):237.6±25.8 vs. 37.1±19.1, EMPs (cells/μL): 28.6±1.8 vs. 5.9±1.8, all P < 0.01]. It was shown by correlation analysis that EMPs were positively related with IL-1β, IL-6, and TNF-α (r value was 0.968, 0.932, 0.945, respectively, all P = 0.000). It was shown by fitting linear regression analysis that when EMPs increased by 1 cell/μL, IL-1β increased by 2.4 ng/L [95% confidence interval (95%CI) = 1.9-2.8, P < 0.001], IL-6 increased by 34.5 ng/L (95%CI = 25.1-44.0, P < 0.001), and TNF-α increased by 13.6 ng/L (95%CI = 10.3-16.9,P < 0.001). It was shown by light microscope that the structure of lung tissue and alveolar of mice in spontaneous breathing group appeared normal, while the shrinks of alveolar and disappearance of alveolar architecture were found in MV group. It was shown by electron microscopy that alveolar wall edema and thickening and broken alveolar septa were found in MV group, by contrast, the structure of alveolar was normal in spontaneous breathing group. Conclusion 30 mL/kg VT ventilation for 4 hours could induce VILI with increase in EMPs, suggesting EMPs closely related to VILI, and EMPs level may be putative biomarker of VILI.
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Objective To evaluate the influences of high hydrostatic pressure (HHP) on the virulence and antigenicity of Coxsackie virus B3 (CB3 V) , to explore a new physiological method to develop an active vaccine against Coxsackie virus. Methods The phagocytosis of the macrophages in mice to the CB3 V and the TCID50 of the CB3V before and after treated by HHP were examined. Results TCID50 increased from 10-5 to 10-4 when treated by 680 MPa; viruses were inactivated at 700 MPa. Viruses were also inactivated by the pressure less than 700 MPa, but needed prolonged time. The phagocytic function, antibody and T lymphocyte transformation rate in experimental mice (heated, attenuated, and inactive viruses) exhibited a significant difference compared with control mice (P
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Objective To study the optimum procedure of extracting ginsenoside from Panax ginseng by super pressure extracting technique (SPET). Methods SPET was used to extract ginsenoside from P. ginseng at the normal temperature. The optimum extracting process was selected by the orthogonal test. The contents of ginsenoside in P. ginseng were determined by UV-spectrophotometry. Results The optimum condition of SPET was as follows: when the solid dissolved in 50% ethanol, ratio of raw P. ginseng (g) and solvent (mL) was 1∶75, the pressure was kept at 500 MPa for 2 min; the high yield of ginsenoside was up to 7.76%. Conclusion SPET has a series of advantages, such as higher efficiency, shorter extracting time, and lower exhausting energy, at the same time, the impurity is little, etc.
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AIM: To study the optimum technological parameters for extraction of Astragalus polysaccharides by ultra-high pressure extraction. METHODS: Measured concentration of total astragalus polysacharides served as the marker,the design of uniform distribution was applied to evaluating the ultra-high pressure extration efficiency in comparison with reflux and ultrasonic extraction efficiency. RESULTS: The optimal parameters of ultra-high pressure extraction was as follows: 350 MPa pressure,60 mL/g ratio of solvent to solid material,5 h soaking time,2 min extraction time.Under the conditions,the high yield was up to 24.2 %. CONCLUSION: Ultra-high pressure extraction has a series of advantages,such as higher yield,less impurity,easier separation-purification,shorter extraction time and lower energy input.