Objective To evaluate the effect of edaravone on apoptosis in hippocampal cells in a rat model of endotoxic shock.Methods Thirty-six male Sprague-Dawley rats, weighing 200-250 g, aged 6 weeks, were randomly divided into 3 groups (n=12 each) using a random number table: control group (group C), endotoxic shock group (group ES), and edaravone group (group E).Lipopolysaccharide 10 mg/kg was injected via the femoral vein to establish the model of endotoxic shock in ES and E groups, while the equal volume of normal saline was given in group C.In group E, edaravone 3 mg/kg was intravenously injected immediately after establishment of the model once every 2 h until the animals were sacrificed.The equal volume of normal saline was given instead of edaravone in C and ES groups.At 6 and 12 h after administration of edaravone, 6 rats in each group were sacrificed, and the hippocampi were isolated for determination of malondialdehyde (MDA) content (using thiobarbituric acid method) , tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) contents (using enzyme-linked immunosorbent assay), and cell apoptosis in hippocampal CA1 region (by TUNEL assay).The apoptotic index was calculated.Results Compared with group C, the MDA, TNF-α and IL-6 contents were significantly increased at 6 and 12 h after administration of edaravone, and the apoptotic index was increased at 12 h after administration of edaravone in ES and E groups.Compared with group ES, the MDA, TNF-α and IL-6 contents were significantly decreased at 6 and 12 h after administration of edaravone, and the apoptotic index was decreased at 12 h after administration of edaravone in group E.Conclusion Edaravone can reduce apoptosis in hippocampal cells, and the mechanism is associated with the reduced oxidative stress and inflammatory responses in a rat model of endotoxic shock.

Objective To evaluate the effects of edaravone on the permeability of blood-brain barrier in septic rats.Methods Ninety male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n=30 each):control group (group C),sepsis group (group lipopolysaccharide (LPS)) and edaravone group (group E).Sepsis was induced by injection of LPS 10 mg/kg via the femoral vein in LPS and E groups.After LPS injection,edaravone 3.0 mg/kg was injected intravenously every 2h for 7 times in group E.The equal volume of normal saline was administered instead of edaravone in C and LPS groups.At 2,6 and 12h after LPS injection,5 rats were chosen and Evan's blue (EB) was injected via the femoral vein,and then the rats were sacrificed and brain tissues were removed for determination of EB and water contents.Another 5 rats were chosen and blood samples were taken from the femoral artery for measurement of serum MDA concentration,and then the rats were sacrificed and the brain tissue was harvested for microscopic examination.Results Compared with group C,brain water and EB contents were significantly increased at 6 and 12h after LPS injection,and the serum MDA concentration was increased at 2,6 and 12h after LPS injection in LPS and E groups (P ＜ 0.05).Compared with group LPS,brain water and EB contents were significantly decreased at 6 and 12h after LPS injection,and serum MDA concentrations were decreased at 2,6 and 12h after LPS injection in group E (P ＜ 0.05).Sepsis-induced pathological changes were significantly attenuated in group E.Conclusion Edaravone can decrease the permeability of blood-brain barrier,attenuate brain edema and brain injury in septic rats,and reduction of oxygen free radical production may be involved in the mechanism.

Objective To investigate the effects of preoperative sleep disturbance on the efficacy of flurbiprofen for postoperative analgesia in patients undergoing endoscopic nasal surgery.Methods Ninety-six ASA Ⅰ or Ⅱ patients of both sexes (aged 20-60 years and weighing 50-80 kg) undergoing endoscopic nasal surgery were enrolled in this study.Pittsburg sleep quality index was used to evaluate the long-term sleep quality before hospitalization and Athens sleep quality index was used to evaluate the short-term sleep quality in hospital.The patients were divided into four groups according to the types of preoperative sleep disturbance (n =24 each):no sleep disturbance (group Ⅰ),long-term sleep disturbance (group Ⅱ),acute short-term sleep disturbance (group Ⅲ),and long-term + acute short-term sleep disturbance (group Ⅳ).Anesthesia was induced with sufentanil,propofol and cis-atracurium and maintained with intravenous infusion of remifentanil and propofol.Then the patients received endotracheal intubation and mechanical ventilation.The end-tidal pressure of carbon dioxide was maintained at 30-35 mm Hg.Controlled hypotension was performed with nicardipine,and the mean arterial blood pressure was maintained at 50-70 mm Hg and heart rate at 60-90 bpm during operation.The patients received intravenous injection of flurbiprofen 50 mg 15 minutes before the end of operation for postoperative analgesia.When the visual analogue scale score was more than 3 during the first 6 hours after operation,flurbiprofen 50 mg was given intravenously as rescue analgesia.Results The incidence of rescue analgesia administered after operation was significantly greater in groups Ⅱ,Ⅲ and Ⅳ than in group Ⅰ,and greater in group Ⅳ than in groups Ⅱ and Ⅲ.There was no significant difference in the incidence of rescue analgesia administered during the first 6 hours after operation between groups Ⅱ and Ⅲ.Conclusion Preoperative sleep disturbance has adverse effects on the efficacy of flurbiprofen for postoperative analgesia in patients undergoing endoscopic nasal surgery.

Objective To evaluate the effects of volume resuscitation with different fluids on expression of aquaporin 1 (AQP-1) in pulmonary capillary endothelial cells of endotoxemic rats.Methods Fifty pathogen-free male Sprague-Dawley,rats,weighing 250-300 g,aged 6-8 weeks,were randomly divided into 4 groups (n =10 each):control group (group C),lipopolysaccharide group (group LPS),NaCl group (group NS),6％ hydroxyethyl starch 130/0.4 group (group HES) and hypertonic hydroxyethyl starch 40 group (group HSH).Sepsis was induced with LPS 5 mg/kg injected via the femoral vein.At l0 min after the model was successfully established,the corresponding fluids were infused into the femoral vein at a constant speed (15 ml/kg over 6 h) via a pump.At 0,3 and 6 h after LPS administration,blood samples were collected from the femoral artery for measurement of serum tumor necrosis factor-α (TNF-α) concentrations and for blood gas analysis.PaO2 and lactate (Lac) concentrations were recorded and oxygenation index (PaO2/FiO2) was calculated.The rats were sacrificed at 6 h after LPS administration,and lungs were removed for determination of AQP-1 expression in pulmonary capillary endothelial cells and wet-to-dry lung weight ratio (W/D ratio),and for microscopic examination of the pathological changes which were scored.Results Compared with group C,W/D ratio,serum TNF-α and Lac concentrations,and pathological scores were significantly increased,AQP-1 expression was down-regulated,and PaO2 and oxygenation index were decreased in LPS,NS,HES and HSH groups.Compared with group LPS,W/D ratio,serum TNF-a and Lac concentrations,and pathological scores were significantly decreased,AQP-1 expression was up-regulated,and PaO2 and oxygenation index were increased in NS,HES and HSH groups.Compared with group NS,W/D ratio,serum TNF-α and Lac concentrations,and pathological scores were significantly decreased,AQP-1 expression was up-regulated,and PaO2 and oxygenation index were increased in HES and HSH groups.Compared with group HES,the blood Lac concentrations were significantly decreased,and no significant changes were found in the other parameters in group HSH.Conclusion Volume resuscitation with NaC1,hydroxyethyl starch 130/0.4 and hypertonic hydroxyethyl starch 40 can reduce the lung injury effectively in endotoxemic rats,and hypertonic hydroxyethyl starch 40 provides more significant efficacy,and up-regulated AQP-1 expression in pulmonary capillary endothelial cells is involved in the mechanism.

BACKGROUND:Gene therapy has become a new trend for disease therapy and brought promise for some refractory diseases. The key point is to choose the proper cell, gene and vector. OBJECTIVE:To investigate the effectiveness and feasibility of bone morphogenetic protein 2 (BMP2) gene transfected into endothelial progenitor cells (EPCs) from rat bone marrow for gene therapy. METHODS:The EPCs were isolated, cultured and identified from the bone marrow of Sprague-Dawley rats. Empty vector (LV-eGFP) or BMP2 gene (LV-eGFP-BMP2) was transferred into EPCs by the constructed lentiviral vector (LV). We examined the transfection efficiency by eGFP fluorescence, BMP2 secretion by ELISA, BMP2 expression by Western blot, and compared the capacities of migration, proliferation and anti-apoptosis after transfection in the three groups of normal EPCs, empty vector-EPCs, and BMP2-EPCs. RESULTS AND CONCLUSION:The transfection efficiency of lentiviral vector was 90%. BMP2 gene-transferred EPCs secreted and expressed more BMP2 proteins (P<0.01), and showed enhanced anti-apoptotic ability (P<0.05). The proliferation and migration capacity did not change obviously (P>0.05). After successful transfection with lentivirus-BMP2 gene, EPCs can secrete and express more BMP2 protein and show enhanced anti-apoptotic ability without obvious influence on the proliferation and migration capacity.

Objective To test whether sodium arsenite can induce in vitro reverse mutation of Salmonella typhimurium histamine-auxotroph mutant. Methods Ames test was carded out with Salmonella typhimurium strains TA97,TA98,TA100 and TA102 by standard method with or without the liver microsomal enzyme activation system (+S9,-S9). Results At concentrations of sodium arsenite from 500.00 to 5000.00 μg/plate, no colonies were seen on the plates of TA97,TA98,TA100 or TA102, with or without the presence of S9. At concentrations of sodium arsenite of 0.01,0.10,10.00 μg/plate and with the presence of S9, twice as many colonies grew on the plates of TA102 as the negative control(P<0.05). Without S9 activation,twice as many colonies grew on the plates of TA100 as the negative control(P<0.05)at concentrations of sodium arsenite of 1.00,10.00 μg/plate(P<0.05). The reverse mutation colonies induced by sodium arsenite in TA98 strain were twice as many as negative control group at concentrations of 0.01,0.10 μg/plate(P<0.05). There was no obvious increase of the strain clones in the other(P0.05). Conclusions With and without S9 activation, the doses of 500.00,5000.00 μg/plate sodium arsenite resulted in a toxic effect and a reduction of the revertants among the strain. At concentrations of 0.01～10.00 μg/plate, sodium arsenite exhibited mutngenesis effects.

Objective To Observe the effects of tumor necrosis factor ? (TNF ?) on cell proliferation and Intracellular free calcium concentraction ([Ca 2+ ]i) in endothelium of human umbilical vein endothelial cells (HUVEC) and investigate the pathogenesis of pregnancy induced hypertension syndrome (PIH). Methods Confluent monolayer of HUVEC was directly incubated with TNF ? at following final concentrations: 500, 1 000, 2 000 U/ml for 24 hours. The percentages of different cellcycles and [Ca 2+ ]i were measured by flow cytometry and fluorospectrophotometry. Results Incubated with TNF ?, the endothelial cells were elongated and transformed into fibroblast like cells. Border of nucleus was sharp, clarity, and cells were in regular shape. But there were abnormal granules in cytoplasma and some cells detached at the concentrotion of 2 000 U/ml of TNF ?. Stimulated by TNF ?, the percentage of cellcycles from phase G 0+G 1 to S and G 2+M decreased significantly and it was dose dependent. [Ca 2+ ]i increased significantly and dose dependent as well. Conclusion TNF ? may injure endothelium directly and inhibit its proliferation and repair through the changes of [Ca 2+ ]i level. It may play an important role in the pathogenesis of PIH.

Adult ; Humans ; Male ; Middle Aged ; Phosphines ; poisoning ; Young Adult

Objective To investigate the effects of dexmedetomidine on lipopolysaccharide (LPS)-induced brain injury in rats.Methods Thirty-six pathogen-free Sprague-Dawley rats,aged 6 weeks,weighing 200-250 g,were randomly divided into 3 groups (n =12 each):control group (group C),LPS group (group L) and dexmedetomidine group (group D).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 350 mg/kg,tracheostomized and mechanically ventilated.Dexmedetomidine 100 μg/kg was injected intraperitoneally and LPS 7.5 mg/kg was injected via the femoral vein 15 min later in group D.Normal saline 2 ml was injected intraperitoneally and LPS 7.5 mg/kg was injected via the femoral vein 15 min later in group L.Normal saline 2 ml was injected intraperitoneally and then injected via the femoral vein 15 min later in group C.Blood samples were obtained from the femoral artery at 2 and 4 h after LPS administration for determination of serum TNF-α concentration by ELISA.Six rats were chosen at 12 h after LPS administration,Evan's blue (EB) was injected via the femoral vein,and then the rats were sacrificed and brains removed for determination of EB content.Another six rats were sacrificed and their brains were immediately removed for determination of brain water content and for microscopic examination.Results The brain water content,EB content and serum TNF-α concentration were significantly increased in groups L and D as compared with group C (P ＜ 0.05).The brain water content,EB content and serum TNF-α concentration were significantly lower in group D than in group L (P ＜ 0.05).The microscopic examination showed that brain injury was attenuated in group D compared with group L.Conclusion Dexmedetomidine can reduce LPS-induced brain injury and reduction of the inflammatory response in the brain tissues and improvement in the permeability of the blood-brain barrier may be involved in the mechanism.

Objective: To investigate the relationship between the plasma concentration of nitric oxide (NO) and prognosis of the hemorrhagic shock. Method:The blood levels of NO and Lactate (LA) were measured with fluorophotometry and colorimetry in 30 hemorrhagic shock patients,and another 30 patients for elective surgery served as a control. Result :Concentration of NO was significantly lower and that of LA was significantly higher in hemorrhagic shock group than that of control group. NO level had a negative correlation with LA level and injury index. NO level in the patients complicated by sepsis were still lower than the control. Conclusion:Decrease of NO level may result in disturbance of microcirculation and increase of LA. So nitroglycerin should be used as early as possible in the hemorrhagic shock patients.