Objective To investigate the effects of ketamine on endotexin-induced production of proinflammatory cytokines(IL-6, TNF-?) and activation of their medulating factor NF-?B in vivo. Methods Forty adult male SD rats weighing 200-250 g were randomly divided into 4 groups: (Ⅰ)control group(n=10); (Ⅱ) endotoxin group received intravenous endotoxin(Escherichia coli O111: B4, Sigma) 5 mg?kg~(-1)(n=10); (Ⅲ, Ⅳ)endotexin+ketamine group received ketamine or 5 or 10mg?kg~(-1)?h~(-1) after endotoxin(n=10). The animals were anesthetized with urethane i.p. (1g?kg~(-1)). Carotid artery was cannulated for BP and HR monitoring and jugular vein was cannulated for fluid or drug administration. Two hours after endotoxin administration the animals were sacrificed by exsanguination. Blood was collected and peripheral blood monocytes(PBMC) were isolated. NF-?B activity in PBMC was measured by EMSA and plasma TNF-? and IL-6 levels were determined by ELISA. Results Progressive hypotension and tachycardia developed after endotoxin administration. Endotexin also increased NF-?B activity in PBMCs and plasma TNF-? and IL-6. Ketamine 10 mg?kg~(-1) attenuated the endotexin-induced hemedynamie levels. Ketamine(5, 50 mg?kg~(-1)?h~(-1)) suppressed NF-?B activity in PBMC and inhihited plasma TNF-? level but plasma IL-6 level was not affected. Conclusion Ketamine can suppress endotoxin-induced NF-?appa B activation. Subanesthetic dose of ketamine has anti-inflammatory action.

Objective To investigate the effect of electro-acupunctare at zusanli on acute lung injury in a rat model of sepsis after scald.Methods Fifty SPF male Sprague-Dawley rats,weighing 200-250 g,aged 2-3 months,were randomly divided into 5 groups (n =10 each) using a random number table:control group (group C),sepsis after scald group (group SS),electro-acupuncture at zusanli group (group E),electric stimulation of non-acupoint group (group NE) and electro-acupuncture at zusanli + α-bungarotoxin (α-BGT,a selective α7 nicotinic acetylcholine receptor antagonist) group (group α-BGT).The rats were subjected to a third degree scald covering 20％ total body surface (TBS) and muramyl dipeptide (MDP) 5 mg/kg was injected into the femoral vein immediately after scald to induce sepsis.Electro-stimulation (3 V,2 ms,3 Hz) of bilateral zusanli was performed for 12 min starting from the time point immediately after MDP injection and every 8 h for 2 consecutive days in group E.In group NE,electro-stimulation was performed at the points 5 mm lateral to the bilateral acupoints of Zusanli and the method was similar to those previously described in group E.In group α-BGT,α-BGT 1.0 μg/kg (in 1 ml of normal saline) was injected into the femoral vein before electro-stimulation of zusanli.At 48 h after treatment,arterial blood samples were obtained for determination of serum tumor necrosis factor-alpha (TNF-α) and high mobility group box-1 (HMGB1) protein levels (by ELISA) and lung specimens were removed for microscopic examination and for determination of the expression of Nod like receptor 2 (NLR2) mRNA (by RT-PCR) and receptor interacting protein 2 (RIP2) in the lung tissues (by Western blot).Results Compared with group C,the expression of NLR2 mRNA and RIP2 was significantly up-regulated,and the serum TNF-α and HMGB1 levels were increased in SS,NE and α-BGT groups (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group E (P ＞ 0.05).Compared with group SS,the expression of NLR2 mRNA and RIP2 was down-regulated,and the serum TNF-α and HMGB1 levels were decreased in group E (P ＜ 0.05),and no significant change was found in the parameters mentioned above in NE and α-BGT groups (P ＞ 0.05).Compared with group E,the expression of NLR2 mRNA and RIP2 was significantly up-regulated,and the serum TNF-α and HMGB1 levels were increased in NE and α-BGT groups (P ＜ 0.05).The pathological changes of lung tissues were significantly reduced in group E as compared with group SS.Conclusion Electro-acupunctare at Zusanli can reduce acute lung injury in a rat model of sepsis after scald and inhibition of NLR2/RIP2 signaling pathway and activation of cholinergic anti-inflammatory pathway in lung tissues may be involved in the mechanism.

Objective To evaluate the effect of penehyclidine hydrochloride on cell apoptosis during acute lung injury (ALI) induced by two-hit in rats.Methods Thirty male SPF Sprague-Dawley rats, aged 8 weeks, weighing 240-270 g, were randomly assigned into 3 groups (n =10 each) using a random number table: sham operation group (group Sham), ALI induced by blunt chest trauma and hemorrhagic shock group (group ALI), and penehyclidine hydrochloric group (group PHC).In ALI and PHC groups, the rats were subjected to the combination of chest trauma and hemorrhage (mean arterial pressure 35-40 mmHg mm Hg, lasting for 60 min) to establish a model of ALI.In group PHC, penehyclidine hydrochloric 2 mg/kg was injected intraperitoneally at 1 h before blunt chest trauma.At 8 h after successful establishment of the model, the rats were sacrificed, and lungs were removed for examination of the pathologic changes and for determination of Bax, Bcl-2 and caspase-3 expression (using Western blot), tumor necrosis factor-alpha (TNF-α) content (by enzyme-linked immunosorbent assay) , and cell apoptosis (by TUNEL).Apoptotic index was calculated.Results Compared with group Sham, the levels of Bax, caspase-3 and TNF-α and apoptotic index were significantly increased, and Bcl-2 expression was down-regulated in ALI and PHC groups.Compared with group ALI, the levels of Bax, caspase-3 and TNF-α and apoptotic index were significantly decreased, and Bcl-2 expression was up-regulated in group PHC.The pathologic changes of lungs were significantly reduced in group PHC than in group ALI.Conclusion Penehyclidine hydrochloride mitigates ALI induced by two-hit through inhibiting cell apoptosis in rats.

Objective To investigate the effect of nicotine on coagulation abnormalities in endotoxemic rats.Methods Ninety-six male SD rats weighing 200-250 g were randomly divided into 4 groups (n=24 each): group normal saline (group NS);group LPS;group nicotine(group NIC)and group α-bungarotoxin (α7 nicotinic acetylcholine receptor antagonist, group α-BGT) . Endotoxemia was induced by LPS 10 mg/kg injected via femoral vein in LPS, NIC and α-BGT groups. In group NIC nicotine 400 μg/kg was injected intraperitoneally at 30 min before LPS injection. In group α-BGT α-BGT 1 μg/kg was injected intraperitoneally at 15 min before intraperitoneal nicotine. Prothrombin time(PT),activated partial thromboplastin time(APTT),fibrinogen(Fib),antithrombin (AT),von Willebrand factor(vWF),plasminogen activator inhibitor-1(PAI-1),D-dimer,platelet count and TNF-α were measured before (baseline) and 2, 4 and 6 h after LPS injection.Results PT and APTT were significantly prolonged and plasma Fib and AT concentrations and platelet count were significantly decreased, while plasma PAI-1, D-dimer, vWF and TNF-α concentrations were significantly increased after LPS administration in group LPS as compared with group NS. Nitotine pretreatment significantly attenuated the LPS-induced changes in group NIC.The effect of nicotine was counteracted by α-BGT. Conclusion Nicotine can attenuate coagulation abnormalities induced by LPS by acting on α7 nicotinic acetylcholine receptor.

The primary structure of rhFK506-Binding Protein (FKBP) 12 was studied with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) by measuring its molecular weight and tryptic peptide mass fingerprinting.The calculated molecular weight of rhFKBP12 was 11819.5 and the measured value was 11818.0. The relative error was only 0.01%. The measured peptide mass fingrprinting of rhFKBP12was identical with that deduced from its amino acid sequence. It proved that the primary structure of rhFKBP12 was correct and there was no amino acid deletion, mutation and modification in its expression, refolding andpurification.

Objective To identify the correlation factors associated with cerebral intragenic ischemia after temporary parent arterial occlusion in intracranial aneurysm surgery. Methods One hundred and eighteen patients who underwent temporary arterial occlusion in the 120 aneurysms (from a group of 324 consecutive aneurysm patients treated from 1996 to 2002) were reviewed retrospectively. These variables included sex, age, presence of preoperative subarachnoid hemorrhage (SAH), neurological clinical grade, operational timing, duration of arterial occlusion, numbers of temporary occlusion, mode of arterial occlusion, intraoperative aneurysm rupture, hypertension, the location of temporary occlusion, aneurysm size, hyperglycemia, atheromatous mass. Univariate and multivariate were used to investigate the relationship between the variates and postoperative ischemic changes. Results The total times of temporary occlusion were 156, with an average of 1.30. The duration of arterial occlusion ranged from 1 to 45 min (9.75?7.75). Seventeen patients (14.4%) demonstrated evidence of new infarction in the vascular territory subjected to temporary arterial occlusion. Conclusion In the univariate analysis, age, presence of preoperative SAH, duration of arterial occlusion, atheromatous mass are all significantly correlated with postoperative ischemic injuries. Multivariate logistic regression revealed that the age, older more than 60 (P= 0.010 3 , relative risk=4.335), and the duration of arterial occlusion, lasting more than 20 min (P= 0.032 9 , relative risk=4.177), have significant correlation with the injuries. Based on these findings, temporary occlusion is safe and useful in aneurysm surgery and the postoperative cerebral ischemia is less likely to occur when the duration of clipping is shorter than 20 min.

Objective:To study the effects of exosomes (EXO) released by adenoid cystic carcinoma SACC-83 cells on the expression of fibroblast activation protein (FAP) in normal human salivary gland stromal fibroblasts (hSGSFs).Methods:ACC exosomes were isolated from SACC-83 cell culture supernatant by using Total Exosome Isolation Reagent.The whole-mount EXO were characterized and assessed by transmission electron microscope and Western Blot.The exosomes were labeled with green fluorescent dye PKH67 and co-cultured with hSGSFs for 48h,followed by staining with Alexa Fluor 594 Phalloidin and DAPI.Mterwards,exsosomes uptake was observed under a laser scanning confocal microscope.After a 48-hour co-culture of SACC-83 exosomes with hSGSFs,the expression of FAP in SACC-83-EXO-treated hSGSFs was investigated by qRT-PCR and Western Blot.Results:The vesicles isolated from SACC-83 cell culture supernatant had the reported size range of 30-100 nm,expressed the exosomal marker CD63 and TSG101.Mter co-culture of hSGSFs with PKH67 labeled SACC-83 exosomes,exosomes were taken up by hSGSFs and FAP expression was elevated in hSGSFs.Conclusion:Exosomes derived from SACC-83 cells can be taken up by hSGSFs and can induce the expression of FAP in hSGSFs.These results suggest that exosomes derived from SACC-83 cells might induce the transformation of normal salivary gland strormal fibroblasts to cancer associated fibroblasts.

Objective To evaluate the regulatory role of acetylcholine receptor in muramyl dipeptide (MDP)-induced activation of Nod-like receptor 2/receptor-interacting protein 2 (2NLR2/RIP2) pathway in macrophages of mice.Methods RAW264.7 cells at the logarithmic growth phase were seeded in 12-well plates (density 1 × 106 cells/ml,2 ml/well),a total of 108 wells.The cells were randomly divided into 3 groups (n =36 each) using a random number table:control group (group C),MDP group (group M),and GTS-21 (a7nAChR specific agonist) group (group G).The cells were routinely cultured in group C.MDP with the final concentration of 10 μg/ml was added to the culture medium in group M.MDP with the final concentration of 10μg/ml and GTS21 with the final concentration of 50 μg/ml were added to the culture medium in group G.The cells were incubated for 24 h.At 1,6 and 24 h of incubation with MDP,12 wells were chosen and the cell suspension was obtained for measurement of NLR2 mRNA expression (by real-time fluorescent quantitative PCR),RIP2 expression (by Western blot),and concentrations of tumor necrosis factor-alpha (TNF-α) and high mobility group box-1 (HMGB1) in the culture media (by ELISA).Results Compared with group C,the levels of NLR2 mRNA,RIP2,TNFα and HMGB1 were significantly increased at each time point in group M (P ＜ 0.05).Compared with group M,the levels of NLR2 mRNA,RIP2,TNF-α and HMGB1 were significantly decreased at each time point in group G (P ＜ 0.05).Conclusion Acetylcholine receptor can suppress MDP-induced transduction of NLR2/RIP2 pathway in macrophages of mice.

Objective To evaluate the effect of electro-acupunctare at Zusanli on liver injury in a rat model of sepsis after scald.Methods Fifty SPF male Sprague-Dawley rats,weighing 200-250 g,aged 2-3 months,were randomly divided into 5 groups (n =10 each) using a random number table:control group (group C); sepsis after scald group (group SS); electro-acupuncture at Zusanli group (group E); electric stimulation of non-acupoint group (group NE); electro-acupuncture at Zusanli + α-bungarotoxin (α-BGT) group (group α-BGT).The rats were subjected to a third degree scald covering 20％ of the total body surface area (TBSA) and sepsis was induced with muramyl dipeptide (MDP) 5 mg/kg injected into the femoral vein immediately after scald.Electro-stimulation (3 V,2 ms,3 Hz) of bilateral Zusanli acupoints or non-acupoints was performed for 12 min every 8 h for 2 consecutive days starting from the time point immediately after MDP was injected in group E.In group NE,electro-stimulation was performed at the points 5 mm lateral to the acupoints of bilateral Zusanli and the method was similar to those previously described in group E.In group α-BGT,α-BGT 1.0 μg/kg (in normal saline 1 ml) was injected into the femoral vein before electric stimulation,then electro-stimulation was performed and the method was similar to those previously described in group E.After 48 h of continuous stimulation,liver specimens were obtained for microscopic examination of the pathological changes.The blood samples were obtained from the abdominal aorta for determination of the levels of serum ALT and AST,tumor necrosis factor-alpha (TNF-α),and high mobility group box-1 (HMGB1) (by ELISA),and expression of Nod-like receptor 2 (NLR2) mRNA (by RTPCR) and receptor-interacting protein 2 (RIP2) in the liver tissues (by Western blot).Results The serum ALT and AST,TNF-α and HMGB-1 levels and NLR2 mRNA and RIP2 expression in liver tissues were significantly higher in SS group than in C group.Compared with SS group,the serum ALT,AST,TNF-α and HMGB1 levels were significantly decreased and NLR2 mRNA and RIP2 expression was down-regulated in E group,and no significant changes were found in the parameters mentioned above in NE group.Compared with E group,the serum ALT,AST,TNF-α and HMGB1 levels were significantly increased,and NLR2 mRNA and RIP2 expression was up-regulated in α-BGT group.The pathological changes in liver tissues were significantly reduced in group EA as compared with SS,NE and α-BGT groups.Conclusion Electro-acupuncture at Zusanli can reduce liver injury in a rat model of sepsis after scald and inhibition of NLR2/RIP2 signaling pathway and activation of cholinergic antiinflammatory pathway mediated by α7 nicotinic acetylcholine receptors in liver tissues may be involved in the mechanism.

Objective To observe the effect of Triclosan( TCS) exposure on Caenorhabditis elegans( c. ele-gans) F1 generation of locomotory behavior, brood size, and generation time. Methods The trial included a control group and 4 TCS treatment groups with different doses (100 nmol/L,1 μmol/L,10μmol/L,20μmol/L),the exposure time being 24 hours,the effect of c. elegans′head thrashes,body bending frequency,the brood size and generation time was observed. Results (1) The control group exposed to 100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,their head thrash frequency of c. elegans F1 was(109. 40±8. 61) times/min,(84. 70±7. 82) times/min,(76. 35±7. 44) times/min,(74. 74±5. 93)times/min,(71. 95±4. 19)times/min,respectively,the head thrash ability of c. elegans was significantly inhibited(F=62. 245,P<0. 01). (2) When the control group was exposed to 100 nmol/L,1 μmol/L,10μmol/L,20 μmol/LTCS,the frequency of c.elegans F1 body bent was (19.94±2.46)times/20 s,(15.13±1.99) times/20 s,(14.63±2.31)times/20 s,(14.69±1.96)times/20 s,(12.00±1.86)times/20 s,respectively,and the comparative differences between groups were statistically significant(F=25. 636,P<0. 01). (3) When the control group was exposed to 0,100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,the body sizes of the c. elegans F1 generation was (286.83±6.01)articles,(273.33±6.41)articles,(214.17±7.25)articles,(173.67±9.20)articles, (118. 50 ± 6. 98 ) articles, respectively, the brood size of the C. elegans F1 generation exposed to 100 nmol/L, 1μmol/L,10 μmol/L,20 μmol/L TCS levels,were reduced by 4. 71%,25. 60%,39. 45%,58. 67%,the ge-neration time of the c. elegans′F1 generation was shortened by 2. 14%-5. 38% in the TCS treatment groups compared with the control group(F=27. 520,P<0. 01). Conclusions After c. elegans exposure to TCS,locomotory behavior can be severe-ly affected,reproductive damage causes a decline in the number of brood size,and the speeding-up of the breeding rate is related to the concentration of TCS concentration-response.