Objective To investigate the influence of platform color on the learning and memory performance of rats in Morris water maze. Methods 36 adult male SD rats were randomized into two groups and put in water mazes with blue platform (blue group, n=18) and white platform (white group, n=18) respectively. The rats were trained 4 times per day for 6 days. The performance was indicated by escape latency in each training. The spatial memory ability of rats was assessed through spatial probe on the 7th day. Results No significant difference of average escape latency was found in rats of two groups in the place navigation in the training on the first day(P＞0. 05). Escape latency on each day of rats in blue group was shorter than that of rats in white group. Significant difference of escape latency was noticed on the 3rd and 4th day between two groups. There was no significant difference in the results of spatial probe in rats of two groups (P＞0. 05). Conclusion The improvement of learning ability in rats of blue group is faster than that in rats of white group.

Objective To evaluate the effects of different degrees of coagulation disorders after surgery on short-term prognosis in patients undergoing cardiac surgery with cardiopulmonary bypass (CPB).Methods A total of 410 patients of both sexes,aged 18-79 yr,with body mass index of 16-28 kg/m2,of American Society of Anesthesiologists physical status Ⅲ or Ⅳ,scheduled for elective cardiac surgery with CPB,were enrolled in the study.After induction of general anesthesia,the patients were tracheally intubated and mechanically ventilated.Combined intravenous-inhalational anesthesia was used.The patients were divided into 3 groups according to the coagulation function at 6 h after surgery:normal function group (n =55),mild disorder group (n =237) and severe disorder group (n =118).Postoperative mechanical ventilation time,duration of intensive care unit stay,length of hospitalization and complications during hospitalization were recorded.Results Compared with normal function group,the length of hospitalization was significantly prolonged,and the rate of delayed discharge from hospital was increased in mild disorder group,and the ventilation time,duration of intensive care unit stay and length of hospitalization were significantly prolonged,the rate of delayed extubation,rate of prolonged intensive care unit stay and rate of delayed discharge from hospital were increased,and the hepatic and nephritic insufficiency and incidence of re-thoracotomy for bleeding were increased in severe disorder group (P＜0.05),and no significant change was found in the incidence of postoperative complications in mild disorder group (P＞0.05).Conclusion For the patients undergoing cardiac surgery with CPB,postoperative mild coagulation disorders exert no effect on short-term prognosis,however,postoperative severe coagulation disorders produce poor prognosis,and correction of severe coagulation disorders should be taken into account.

Objective RY-15, a specific agonist of Sphingosine 1-phosphate Receptor 3, was synthesized for investigating the function and mechanism of S1PR3 in bacterial clearance. Methods Measure the ability of RY-15 with FITC to enter the THP-1 cell after coculture for 5 min, 10 min, 20 min, 30 min through confocal microscopy. The function of GY-5 and RY-15 in bacterial clearance was observed by gentamicin protection test. The phosphorylation level of ERK and p-ERK in THP-1 cell was detected by Western Blot after GY-5 and RY-15 stimulation for different times. Results According to confocal microscopy, RY-15 started to enter the THP-1 cell after stimulating for 10 min and the effect of entering cell was very obvious after stimulating for 30 min. Compared to GY-5 group, live bacteria in the macrophage were largely decreased in the RY-15 group( P<0.05). Conmpared to GY-5 group, the p-ERK level raised largely at different poins. Conclusions RY-15, a specific agonist of Sphingosine 1-phosphate Receptor 3, can promote bacterial clearance through entering cell and the phosphorylation level of ERK is a possible mechanism.

Objective:To evaluate the role of mitophagy in cognitive dysfunction in rats with sepsis-associated encephalopathy (SAE).Methods:Twenty-four clean-grade healthy male Sprague-Dawley rats, aged 13-14 weeks, weighing 230-250 g, were divided into 3 groups ( n=8 each) using a random number table method: sham operation group (Sham group), SAE group and SAE+ autophagy inhibitor 3-methyladenine (3-MA) group (3-MA group).The SAE models were developed by cecal ligation and puncture in anesthetized animals.3-MA 10 mg/kg was intraperitoneally injected at 30 min after developing the model in 3-MA group.Cognitive function was assessed by Morris water maze test, and the escape latency and ratio of the time of staying at the target quadrant were recorded.After the end of Morris water maze test, the rats were sacrificed and hippocampal tissues were collected for microscopic examination of the pathological changes which were scored after hematoxylin-eosin staining and for determination of the expression of autophagy-related proteins LC3, Beclin1 and p62 (by Western blot).The ratio of LC3Ⅱ/LC3Ⅰwas calculated.The hippocampal mitochondria were isolated to measure mitochondrial membrane potential (MMP), ATP content and ATPase activity by spectrophotometry. Results:Compared with Sham group, the escape latency was significantly prolonged, the ratio of the time of staying at the target quadrant was decreased, the pathological score of hippocampus was decreased, and the contents of MMP and ATP and ATPase activity were decreased in SAE and 3-MA groups, the ratio of LC3Ⅱ/LC3Ⅰwas significantly increased, the expression of Beclin1 was up-regulated, and the expression of p62 was down-regulated in SAE group, and the ratio of LC3Ⅱ/LC3Ⅰwas significantly decreased, and the expression of Beclin1 and p62 was up-regulated in 3-MA group ( P<0.05).Compared with SAE group, the escape latency was significantly prolonged, the ratio of the time of staying at the target quadrant was decreased, the pathological score of hippocampus was decreased, the ratio of LC3/LC3Ⅰwas decreased, the expression of Beclin1 was down-regulated, the expression of p62 was up-regulated, and the contents of MMP and ATP and ATPase activity were decreased in 3-MA group ( P<0.05). Conclusions:Hippocampal mitophagy is involved in cognitive dysfunction in the rats with SAE.

Objective To screen the risk factors for blood coagulation abnormality in patients undergoing off-pump coronary artery bypass grafting (OPCABG).Methods A total of 140 patients undergoing elective OPCABG were included in this study,and combined intravenous-inhalational anesthesia was performed during operation.The patients were divided into normal group and abnormal group according to whether or not blood coagulation abnormality developed during operation and within 48 h after operation.The data such as gender,age,body mass index,American Society of Anesthesiologists physical status,the number of operation per year for surgeons,comorbidities (hypertension and diabetes mellitus),preoperative hematocrit (Hct),left ventricular ejection fraction,arterial oxygen pressure,liver function,operation time and requirement for intraoperative continuous cardiac output monitoring,positive end expiratory pressure,tranexamic acid,ulinastatin and hydroxyethyl starch,postoperative acidosis and hypothermia were recorded.Results Blood coagulation abnormality was found in 43 patients,and the incidence was 31％.The results of logistic regression analysis showed that the number of operation per year for surgeons＜ 50,preoperative abnormal liver function,preoperative Hct＜35％,surgery time≥240 min,no use of continuous cardiac output monitoring during operation and postoperative hypothermia were risk factors for blood coagulation abnormality in patients undergoing OPCABG.Conclusion The number of operation per year for surgeons＜50,preoperative abnormal liver function,preoperative Hct ＜ 35％,operation time ≥ 240 min,no use of continuous cardiac output monitoring during operation and postoperative hypothermia are risk factors for blood coagulation abnormality in patients undergoing OPCABG.

Objective To evaluate the effect of tetramethylpyrazine on autophagy in the hippocam-pal neurons of rats with sepsis-associated encephalopathy. Methods Sixty SPF healthy male Sprague-Daw-ley rats, aged 11-14 weeks, weighing 200-240 g, were divided into 3 groups(n=20 each)using a ran-dom number table: sham operation group(group Sham), sepsis group(group Sep)and tetrameth-ylpyrazine group(group TMP). Sepsis was induced by cecal ligation and puncture(CLP), and group Sham only underwent simple laparotomy. Tetramethylpyrazine 10 mg∕kg was injected intraperitoneally at 1 h before CLP in group TMP. Morris water maze test was performed in 10 rats randomly selected at 12 and 36 h after CLP. Then the rats were sacrificed, and hippocampi were isolated for determination of the expres-sion of microtubule-associated protein 1 light chain 3Ⅰ(LC3Ⅰ), LC3Ⅱ, Beclin-1 and p62 in hipp-ocampal tissues by Western blot, and the LC3Ⅱ∕LC3Ⅰratio was calculated. Results Compared with group Sham, the escape latency was significantly prolonged, the rate of time spent in the target quadrant was decreased, the LC3Ⅱ∕LC3Ⅰratio was increased, the expression of Beclin-1 was up-regulated, and the expression of p62 was down-regulated at 12 and 36 h after CLP in group Sep and group TMP(P<005). Compared with group Sep, the escape latency was significantly shortened, the rate of time spent in the target quadrant was increased, the LC3Ⅱ∕LC3Ⅰratio was decreased, the expression of Beclin-1 was down-regulated, and the expression of p62 was up-regulated at 12 and 36 h after CLP in group TMP(P<005). Conclusion The mechanism by which tetramethylpyrazine reduces sepsis-associated encephalopa-thy is related to inhibiting autophagy in the hippocampal neurons of rats.

Objective:To evaluate the effect of tetramethylpyrazine on hippocampal inflammatory responses in rats with sepsis-associated encephalopathy.Methods:Sixty healthy male Sprague-Dawley rats, aged 12-14 weeks, weighing 240-270 g, were divided into 4 groups ( n=15 each) using a random number table method: sham operation group (group Sham), sepsis-associated encephalopathy group (group SAE), low-dose tetramethylpyrazine group (group L-TMP), and high-dose tetramethylpyrazine group (group H-TMP). Sepsis-associated encephalopathy was induced by cecal ligation and puncture (CLP) in anesthetized rats.Tetramethylpyrazine 5 and 20 mg/kg were intraperitoneally injected once a day in L-TMP and H-TMP groups, respectively, at 5 days prior to CLP.Morris water maze test was performed at 1-5 days after CLP to assess the cognitive function, and the escape latency and ratio of time spent in the target quadrant were recorded.Five rats were sacrificed at 1 day after CLP, the brains were removed, and the hippocampi were isolated for determination of the contents of interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and IL-6 by enzyme-linked immunosorbent assay.Rats were sacrificed after the end of Morris water maze test, and hippocampi were removed for detection of the expression of Toll-like receptor 1 (TLR1), activated caspase-3, Bax and Bcl-2 by using Western blot. Results:Compared with group Sham, the escape latency was significantly prolonged, the ratios of time spent in the target quadrant were decreased, the expression of TLR1, activated caspase-3 and Bax was up-regulated, and the expression of Bcl-2 was down-regulated in group SAE, group L-TMP and group H-TMP, and the contents of IL-1β, TNF-α and IL-6 were significantly increased in group SAE and group L-TMP ( P<0.05). Compared with group SAE, the escape latency was significantly shortened, the ratio of time spent in the target quadrant was increased, the contents of IL-1β, TNF-α and IL-6 were decreased, the expression of TLR1, activated caspase-3 and Bax was down-regulated, and the expression of Bcl-2 was up-regulated in group L-TMP and group H-TMP ( P<0.05). Conclusion:The mechanism by which tetramethylpyrazine reduces sepsis-associated encephalopathy may be related to inhibiting hippocampal inflammatory responses in rats.

Objective:To evaluate the role of p38 mitogen-activated protein kinase (MAPK)/cyclic adenosine monophosphate response element-binding protein (CREB) signaling pathway in tetramethylpyrazine-induced reduction of hippocampal inflammatory responses in mice with sepsis-associated encephalopathy (SAE).Methods:Sixty healthy male C57BL6 mice, weighing 24-27 g, were divided into 4 groups ( n=15 each) using a random number table method: sham operation group (group Sham), sepsis group (group Sep), tetramethylpyrazine group (group TMP) and p38 MAPK inhibitor SB203580 group (group SB). The model of SAE was established by cecal ligation and puncture in anesthetized mice.Tetramethylpyrazine 10 mg/kg was injected intraperitoneally once a day at 3 days before the establishment of the model in TMP group, and SB203580 2.0 mg/kg was intraperitoneally injected at 30 min after the establishment of the model in SB group.The equal volume of normal saline was given intraperitoneally in Sham and Sep groups.At 1 day after operation, cognitive function was assessed by Morris water maze, and the escape latency and ratio of time spent in the target quadrant were recorded.The animals were sacrificed after the test, and hippocampal tissues were taken for determination of the contents of interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and IL-6 (by enzyme-linked immunosorbent assay) and for detection of the expression of phosphorylation of p38 MAPK, GSK3 and CREB and expression of brain-derived neurotrophic factor (BDNF) (by Western blot). Results:Compared with group Sham, the escape latency was significantly prolonged, the ratios of time spent in the target quadrant were decreased, the contents of IL-1β, TNF-α and IL-6 were increased, the phosphorylation of hippocampus p38 MAPK was increased, the phosphorylation of GSK3 and CREB were decreased, and the expression of BDNF was down-regulated in Sep, TMP and SB groups ( P<0.05). Compared with group Sep, the escape latency was significantly shortened, the ratios of time spent in the target quadrant were increased, the contents of IL-1β, TNF-α and IL-6 were decreased, the phosphorylation of hippocampus p38 MAPK was decreased, the phosphorylation of GSK3 and CREB were increased, and the expression of BDNF was up-regulated in TMP and SB groups ( P<0.05). Compared with group TMP, no significant change was found in the parameters mentioned above in group SB ( P>0.05). Conclusion:p38 MAPK/CREB signaling pathway is involved in the process of tetramethylpyrazine-induced reduction of hippocampal inflammatory responses in mice with SAE.

Objective:To design a modified S1PR3 specific agonist, GPS-725.017, and investigate its protective effect on acute lung injury by promoting macrophage clearance of bacteria.Methods:A short peptide derived from the intracellular region of S1PR3 receptor was named GPS725.017, which was modified with norleucine (Nle) and myristicacid (myr) at its N terminus. Mice were divided into the sham operation group, solvent group and GPS-725.017 treatment group. The acute lung injury model was induced by endotracheal injection of E. coli (5×10 6 CFU), and the experimental group was treated with GPS-725.017 (10 mg/kg). The 48-h survival rate of mice was recorded. After 5 h of modeling, the bacterial load and inflammatory cytokines in peripheral blood and lung were detected, and Vps34 protein content in alveolar macrophages was determined by Western blot. After 12-h of modeling, lung tissues were collected for H&E staining and pathological scores. Results:Compared with the solvent group, the survival rate of mice in the GPS-725.017 treatment group was significantly improved ( P<0.01), the bacterial CFU in blood and alveolar lavage fluid was significantly lower than that in the solvent group ( P<0.001), and the levels of TNF-α and IL-1β in blood and alveolar lavage fluid were significantly lower than those in the solvent group ( P<0.001). Western blot showed that the expression level of Vps34 protein in alveolar macrophages was significantly higher than that in the solvent group ( P<0.01). Histopathology result showed that the pathological damage of lung in the treatment group was significantly less than that in the solvent group ( P<0.001). Conclusions:The modified synthetic S1PR3 specific agonist GPS-725.017 could specifically activate the S1PR3 receptor on the membrane of alveolar macrophages and up-regulate the expression level of intracellular Vps34 protein, which can promote the removal of bacteria in alveolar macrophages, significantly reduce the degree of lung injury and improve the survival rate in ALI mice.