Objective The purpose of this study was to investigate the role of p38 mitogen-activated protein kinase ( p38 MAPK) in the lung injury induced by mechanical ventilation.Methods Fifteen healthy 80 day-old pigs weighing (22.5　? 1.5)kg were randomly divided into three groups according to the tidal volume(VT) and PEEP of mechanical ventilation: group A (VT = 16ml?kg-1, PEEP = 0) ; group B (VT = 6 ml?kg-1, PEEP= 16cm H2O) and group C(VT = 16ml?kg-1, PEEP = 8cm H2O). The animals were mechanically ventilated for 3h, then sacrificed by exsanguination. Right lower lobe was immediately removed for identification of intercellular adhesion molecule-1 ( ICAM-1 ) expression using immunohistological technique, determination of phosphorylated p38 MAPK content using Western Blot and microscopic examination. Results There was significant histological changes in the lung tissue in group A and B, but no significant histological changes were found in group C. The expression of ICAM-1 was positive in the lung in group A and B but negative in group C. The level of phosphorylated p38 MAPK among the 3 groups. Conclusion Acute lung injury can be induced by mechanical ventilation with high tidal volume or low tidal volume plus high PEEP, p38 MAPK may mediate the inflammatory response-induced lung injury.

Objective To evaluate the effects of limited fluid resuscitation on systemic inflammatory responses in rats with traumatic hemorrhagic shock through comparing with unlimited fluid resuscitation.Methods Sixty pathogen-free male Sprague-Dawley rats,aged 2-3 months,weighing 250-290 g,were randomly divided into 6 groups (n =10 each) using a random number table:sham operation group (group S),no fluid resuscitation group (group NF),unlimited fluid resuscitation group (group ULF),limited crystalloid fluid resuscitation group (group LR),and limited colloid fluid resuscitation groups (group LSG and group LHES).Traumatic uncontrolled hemorrhagic shock was induced by withdrawal of blood from the femoral artery at 2.5 mL/100 g over a 20-minute period,followed by tail amputation at 10 min after the end of blood withdrawal.At 10 min after the end of blood withdrawal,fluid resuscitation was performed.Lactated Ringer's solution (ULF and LR groups),4 ％ succinylated gelatin (group LSG),or 6 ％ hydroxyethyl starch 130/0.4 (group LHES) was infused intravenously.The initial infusion rate was 2 ml · kg-1 · min-1.The target MAP was maintained at 50 mm Hg in rats with limited fluid resuscitation,while at 80 mm Hg in rats with unlimited fluid resuscitation.After 60 min of fluid resuscitation,bleeding in the tail was stopped by ligation and fluid infusion was replaced with blood resuscitation.After 60 min of blood resuscitation,180 main of observation was started.At 10 min after catheterization of the femoral artery and vein (T0),10 min after the end of blood withdrawal (T1),the end of fluid resuscitation (T2),the end of blood resuscitation (T3),and the end of observation (T4),arterial blood samples were collected to measure hematocrit (Hct)and concentrations of plasma tumor necrosis factor-alpha (TNF-α),interleukin (IL)-6,and IL-10.Blood samples were collected from the femoral artery at T2 for determination of the expression of Toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88) and activity of nuclear factor-kappaB (NF-κB) in monocytes.The amount of blood loss from the tail and volume of fluid infused were also recorded.Another 120 Sprague-Dawley rats were randomly divided into 6 groups (n =20 each) and resuscitation was performed according to the method previously described.The rats were observed for 72 h survival rate.Results Compared with group S,Hct was significantly decreased,the concentrations of plasma TNF-α,IL-6,and IL-10 and activity of NF-κB were increased,and the expression of TLR4,and MyD88 in monocytes was up-regulated in the other groups (P ＜ 0.05).Compared with group NF,the concentrations of plasma TNF-α and IL-6 and NF-κB activity were significantly increased,and the concentration of plasma IL-10 and Hct were decreased,and the expression of TLR4 and MyD88 in monocytes was up-regulated in ULF,LR and LSG groups,and the concentrations of plasma TNF-α and IL-6 were significantly increased,the concentration of plasma IL-10 and Hct were decreased in group LHES (P ＜ 0.05).Compared with group ULF,the concentrations of plasma TNF-α and IL-6 and NF-κB activity were significantly decreased,the concentration of plasma IL-10 and Hct were increased,the survival rate was higher,the expression of TLR4 and MyD88 in monocytes was down-regulated,and the amount of blood loss from the tail was decreased and the volume of fluid infused was reduced in LSG,LHES and LR groups (P ＜ 0.05).Compared with group LR,the concentrations of plasma TNF-α and IL-6 and NF-κB activity were significantly decreased and the expression of TLR4 and MyD88 in monocytes was down-regulated (P ＜ 0.05),and no significant change was found in the concentration of plasma IL-10 in group LHES (P ＞ 0.05),and the volume of fluid infused was reduced and the survival rate was increased (P ＜ 0.05),and no significant change was found in the amount of blood loss from the tail in LSG and LH-ES groups (P ＞ 0.05).Conclusion Compared with unlimited fluid resuscitation,limited fluid resuscitation exerts less effect on systemic inflammatory responses in rats with traumatic hemorrhagic shock,especially when resuscitation with 6％ hydroxyethyl starch 130/0.4 is performed,and inhibition of TLR4/NF-κB signaling pathway is involved in the mechanism.

Objective To observe the differences of biological property of glioma stem cells (GSCs) and glioma non-stem cells (nGSCs), and their related protein expressions. Methods The proliferations of GSCs1, GSCs2 and nGSCs1 and nGSCs2 were detected by CCK8 after two, 4, 6, 8, 10 and 12 d of culture in vitro. The sensitivities of the cells to temozolomide (TMZ) were detected by CCK8 after 2 d of culture. The adhesion abilities of cells were tested by adhesion assay. Transwell assay was used to detect the migration and invasion abilities of cells. The activity of matrix metalloproteinase-2 (MMP-2) was detected by gelatin zymography. Western blotting and immunofluorescence staining were used to detect the protein expressions of Notchl and epidermal growth factor receptor (EGFR). Results The survival rate of nGSCs1 was significantly higher than that of GSCs1 and the survival rate of nGSCs2 was significantly higher than that of GSCs2 after 4, 6, 8, 10 and 12 d of culture (P<0.05). The inhibitory concentration (IC)50 of TMZ for GSCs1, nGSCs1, GSCs2 and nGSCs2 was (1536.0±17.67) μmol/L, (514.5±13.44) μmol/L, (2543.0±39.87) μmol/L, (889.6±17.43) μmol/L, respectively (P<0.05). Number of GSCs1 adhering to extracellular matrix proteins Fibronectin and Collagen I was significantly larger than that of nGSCs1, and that of GSCs2 was significantly larger than that of nGSCs2 (P<0.05). The number of migrated GSCs112 and 24 h of cultivation was statistically larger than that of nGSCs1, and that of GSCs2 was statistically larger than that of nGSCs2 (P<0.05). The number of invaded GSCs124 and 36 h of cultivation was larger than that of nGSCs1, and that of invaded GSCs2 was larger than that of nGSCs2, with statistical differences (P<0.05). The activity of MMP2 secreted by GSCs1 was significantly higher than that by nGSCs1, and that of MMP2 secreted by GSCs2 was significantly higher than that by nGSCs2 (P<0.05). Western blotting showed that the relative protein expression level of EGFR/Notch1 in GSCs1 was significantly lower than that in nGSCs1, and that in GSCs2 was significantly lower than that in nGSCs2 (P<0.05). The results of immunofluorescence staining were consistent with those of Western blotting; EGFR protein strongly expressed in nGSCs and weakly expressed in GSCs; Notch1 protein strongly expressed in GSCs and weakly expressed in nGSCs. Conclusion As compared with the high-EGFR-expressing and proliferative primary glioma cells, the high-Notch1-expressing glioma stem cells have higher activity level of MMP-2,stronger abilities of adhesion, migration and invasion, which may be contributed to glioma treatment resistance and its occurrence.

ObjectiveTo investigate the therapeutic effect of sequential administration of Dihuang Baoyuan granules （DHBY， the prescription for consolidating body resistance） and Fuling Yunhua granules （FLYH， the prescription for treating symptoms） on spontaneous type 2 diabetes mellitus （T2DM） in mice. MethodsAccording to the fasting blood glucose （FBG） level， 12-week-old db/db mice were randomized into six groups： model， DHBY （18.02 g·kg^-1）， FLYH （14.80 g·kg^-1）， sequential administration 1 （SEQ-1， DHBY 18.02 g·kg^-1+FLYH 14.80 g·kg^-1）， sequential administration 2 （SEQ-2， FLYH 14.80 g·kg^-1+DHBY 18.02 g·kg^-1）， and dapagliflozin （Dapa， 1.3 mg·kg^-1）. The m/m mice in the same litter were selected as the normal group. The mice were administrated with corresponding drugs by gavage for 8 consecutive weeks. During the 8 weeks of drug administration and 2 weeks after withdrawal， the retinal thickness， FBG， hemoglobin A1c （HbA1c）， and insulin were determined， and histopathological changes of the pancreas， liver， kidney， and retina were observed by hematoxylin-eosin （HE） staining. ResultsCompared with the model group， SEQ-1 for 4 weeks lowered the FBG level （P<0.05）， raised the insulin level， decreased the triglyceride （TG） level （P<0.05）， increased the number of optic ganglion cells and diminished vacuolar degeneration of pancreatic islet and liver. SEQ-2 lowered FBG and HbA1c levels （P<0.05）， rose the insulin level， increased the retinal thickness and the number of optic ganglion cells （P<0.05）， and alleviated vacuolar degeneration of pancreatic islet and liver. Two weeks after drug withdrawal， Dapa tended to increase FBG and HbA1c compared with those at the time of drug withdrawal. However， the levels of FBG and HbA1c in the SEQ-2 group remained decreasing （P<0.05）. ConclusionSEQ-1 and SEQ-2 can lower the blood glucose level and ameliorate diabetic retinopathy， and SEQ-2 outperformed DHBY and FLYH in lowering the blood glucose level. Moreover， SEQ-2 can maintain the blood glucose-lowering effect after drug withdrawal.

ObjectiveTo investigate the therapeutic effect of sequential administration of Dihuang Baoyuan granules （DHBY， the prescription for consolidating body resistance） and Fuling Yunhua granules （FLYH， the prescription for treating symptoms） on spontaneous type 2 diabetes mellitus （T2DM） in mice. MethodsAccording to the fasting blood glucose （FBG） level， 12-week-old db/db mice were randomized into six groups： model， DHBY （18.02 g·kg^-1）， FLYH （14.80 g·kg^-1）， sequential administration 1 （SEQ-1， DHBY 18.02 g·kg^-1+FLYH 14.80 g·kg^-1）， sequential administration 2 （SEQ-2， FLYH 14.80 g·kg^-1+DHBY 18.02 g·kg^-1）， and dapagliflozin （Dapa， 1.3 mg·kg^-1）. The m/m mice in the same litter were selected as the normal group. The mice were administrated with corresponding drugs by gavage for 8 consecutive weeks. During the 8 weeks of drug administration and 2 weeks after withdrawal， the retinal thickness， FBG， hemoglobin A1c （HbA1c）， and insulin were determined， and histopathological changes of the pancreas， liver， kidney， and retina were observed by hematoxylin-eosin （HE） staining. ResultsCompared with the model group， SEQ-1 for 4 weeks lowered the FBG level （P<0.05）， raised the insulin level， decreased the triglyceride （TG） level （P<0.05）， increased the number of optic ganglion cells and diminished vacuolar degeneration of pancreatic islet and liver. SEQ-2 lowered FBG and HbA1c levels （P<0.05）， rose the insulin level， increased the retinal thickness and the number of optic ganglion cells （P<0.05）， and alleviated vacuolar degeneration of pancreatic islet and liver. Two weeks after drug withdrawal， Dapa tended to increase FBG and HbA1c compared with those at the time of drug withdrawal. However， the levels of FBG and HbA1c in the SEQ-2 group remained decreasing （P<0.05）. ConclusionSEQ-1 and SEQ-2 can lower the blood glucose level and ameliorate diabetic retinopathy， and SEQ-2 outperformed DHBY and FLYH in lowering the blood glucose level. Moreover， SEQ-2 can maintain the blood glucose-lowering effect after drug withdrawal.