Objective: To determine if naloxone has reverse effect on cerebral ischemic injury following cardiac arrest. Method:Ten dogs underwent 15-min cardiac arrest and then were divided into two groups during 3-hour cardiopulmonary resuscitation (CPR). Group A(n=5): the animals were administered with conventional CPR;group B(n=5): in addition,at onset of CPR,an I. V. bolus of 0.2mg?kg~(-1) of naloxone,followed by infusion of 0.02mg?kg~(-1)?h~(-1). Result: Cerebral blood flow, Cerebral perfusion pressure in group B were significantly higher than those in group A (P

Objective To evaluate the effect of sevoflurane on the expression of α7 nicotinic acetylcholine receptor (α7nAChR) in hippocampal neurons of rats.Methods Forty-eight pathogen-free healthy adult Sprague-Dawley rats of both sexes,weighing 210-280 g,aged 3-4 months,were divided into 4 groups (n=12 each) using a random number table:control group (group C) and 1%,3% and 5% sevoflurane groups (S1,S2 and S3 groups).In S1,S2 and S3 groups,1%,3% and 5% sevoflurane were inhaled for 1 h,respectively.Morris water maze test was performed at 1 and 7 days after anesthesia to assess the cognitive function.Six rats in each group were sacrificed after the end of the test.The brains were then removed and the hippocampi were isolated for detection of the expression of α7nAChR (by Western blot) and contents of tumor necrosis factor-alpha and interleukin-1 (by enzyme-linked immunosorbent assay).Results Compared with group C,the number of crossing the platform was significantly decreased at 1 and 7 days after anesthesia in group S1,the time of staying at the target quadrant was significantly shortened,the number of crossing the platform was decreased,and the rate of time of staying at the original platform quadrant and rate of swimming distance at the original platform quadrant were decreased at 1 and 7 days after anesthesia in S2 and S3 groups,and the contents of tumor necrosis factor-alpha and interleukin-1 were significantly increased,and the expression of α7nAChR was down-regulated at 1 and 7 days after anesthesia in S1,S2 and S3 groups (P<0.05).Compared with group S1,the expression of α7nAChR was significantly down-regulated at 1 and 7 days after anesthesia in S2 and S3 groups (P<0.05).Compared with group S2,the expression of α7nAChR was significantly down-regulated at 1 day after anesthesia in group S3 (P<0.05).There was no significant difference in each parameter of Morris water maze test among group S1,group S2 and group S3 (P>0.05).Conclusion The mechanism by which sevoflurane induces inflammatory responses in hippocampi is related to down-regulation of α7nAChR expression in hippocampal neurons of rats.

Objective Compare the three kind of criteria in evaluating the incidences of postop-erative cognitive dysfunction in non-cardiac surgery.Methods Four hundred and sixty one non-cardiac surgery patients were randomly enrolled in this study group and two hundred forty four non-surgery patients as control group.Patients??cognitive state was measured on preoperative and postoperative 1 and 3 d by mini-mental state examination(MMSE).POCD was assessed by patients??education level, one standard deviation and Z-score scale,respectively.Results On the first and third day after sur-gery,Z-score scale assess POCD result showed the highest relevance ratio and 95%CI as well as the lowest education level.The education scale showed the lowest incidence of POCD.Education level criteria was positive on postoperative 1 and 3 day,while one case on postoperative 1 day and four cases on postoperative 3 day were negative by Z-score scale.Conclusion The incidence of POCD in same pa-tients by three kind of criteria are different.The Z-score scale is recommended to evaluate the POCD in order to avoid misdiagnosis.

Objective To investigate the accuracy of serum S-100β protein and neuron specific enolase (NSE) level in predication of postoperative delirium (PD) in patients of different ages.Methods Four hundred ASA Ⅰ -Ⅳ patients of both sexes weighing 40-82 kg undergoing abdominal surgery performed under general anesthesia were divided into 4 age groups:group Ⅰ 18-44 yr; group Ⅱ 45-59 yr; group Ⅲ 60-74 yr and group Ⅳ ≥75 yr.The diagnosis of PD was made by using confusion assessment method.The incidence of PD was recorded within 72 h after operation.Each group was further divided into PD and non-PD subgroups.Blood samples were taken at 1 day before operation (T1),during their stay in PACU (T2) and at 24 and 72 h after operation (T3,4 ) for determination of serum S-100β protein and NSE concentrations.The receiver operating characteristic (ROC) curve for serum S-100β protein concentration in determining the PD efficacy was plotted.Results The incidence of PD was significantly higher and the duration was significantly longer in groups Ⅲ and Ⅳ than in groups Ⅰ and Ⅱ,and in group Ⅳ than in group Ⅲ (P ＜ 0.05).There was no significant difference in the serum S-100β protein concentration between PD subgroup and non-PD subgroup in groups Ⅰ - Ⅲ ( P ＞ 0.05).Compared with that at T1 and in nonPD subgroup,the serum S-100β protein concentration was significantly increased in PD subgroup in group Ⅳ,and the serum NSE concentration was significantly decreased at T2,3 in PD subgroup in group Ⅰ (P ＜ 0.05).There was no significant difference in the serum NSE concentration between PD subgroup and non-PD subgroup in groups Ⅱ -Ⅳ.The analysis results of the ROC curve showed that:the area under the curve for the serum S-100β protein concentration and 95％ confidence interval were 0.329 (0.127-0.531),0.352 (0.168-0.536),0.619 (0.466- 0.772) and 0.921 (0.846-0.995),the sensitivity was 50％,50％,56％ and 88％,and the specificity was 29％,22％,46％ and 86％ in groups Ⅰ-Ⅳ respectively.Conclusion Increase in the serum S-100β protein concentration can be used in predicting the development of PD in patients ≥75 yr,but the serum NSE protein concentration can not be used.

AIM: To explore the effect of Snail1 siRNA on high-glucose induced tubular epithelial-to-mesenchymal transition (TEMT). METHODS: Subconfluent renal tubular epithelial cells were incubated in serum-free DMEM for 24 h to arrest and synchronize the cell growth. Then cells were treated with normal glucose (5.5 mmol/L D-glucose) or high glucose (25 mmol/L D-glucose) for 72 h. Meanwhile 19.5 mmol/L D-manntiol was used as high osmotic control. Snail1 siRNA was transfected into tubular epithelial cells. In parallel, cells were transfected with non-specific siRNA which served as the control data sets. Cells were then treated with 25 mmol/L D-glucose for 72 h. RNA and cell lysates were collected to determine the protein and mRNA levels of Snail1, TGF-β_1, α-SMA, vimentin and E-cadherin. RESULTS: Transfection caused the decreases in Snail1 at mRNA and protein levels by 62% and 68% respectively as compared to those in untransfected cells cultured in high glucose medium. Western blotting exhibited that Snail1 siRNA transfection restored E-cadherin protein expression by 61% compared to that in high-glucose-treatment cells, whereas it inhibited high-glucose-induced induction of α-SMA protein by 58%. Similarly, RT-PCR revealed that Snail1 siRNA transfection dramatically suppressed the high-glucose-induced mRNA expressions of α-SMA and vimentin by 72% and 61%, respectively, while E-cadherin mRNA increased by 53%. CONCLUSION: Our study provides direct evidence that Snail1 is able to control TEMT.

BACKGROUND: Meseuchymal stem cells (MSCs) have extremely strong self-duplication ability and multidirectional ifferentiation potential. When bone marrow stromal cells (BMSC) are isolated and cultured in vitro, implanted in vivo, the distribution and colonization are still unclear, which is concerned with whether BMSC can be usedas target cells in clinic.OBJECTIVE: To explore the capacity of colonizing to the liver after allografting of green fluorescent protein (GFP) labeled MSCs of rats by different approaches.DESIGN: Factorial design.SETTING: Department of General Surgery, Second People's Hospital of Guangdong Province, Postdoctoral Workstation of Sun Yat-Sen University;Department of General Surgery, Zhujiang Hospital, Southern Medical University; Department of Organ Transplantation, Third Hospital Affiliated to Sun Yat-Sen University.MATERIALS: The experiment was performed at the Staff Room of Pharmacology, Basic Department, First Military Medical University of Chinese PLA from January 2003 to December 2004. A total of 36 clean adult SD rats were selected and randomly assigned into 5 groups: CCL4 plus portal vein transplantation group (n=6), portal vein transplantation control group (n=6), CCL4 plus caudal vein transplantation group (n=6), caudal vein transplantation control group (n=6) and mixed group (n=12).METHODS: ① MSCs were obtained from rat marrow and labeled with GFP. After amplifying in vitro, MSCs suspension was implanted with thin needle, with the volume of 0.5 mL/100 g. ②CCL4 plus portal vein transplantation group: In 3 days before MSCs transplantation, the rats were administrated with 20 g/L CCL4 2.5 mL/kg by gastric perfusion every day. The dose was double at the first time. Labeled MSCs were implanted from portal vein. Portal vein transplantation control group: Before transplantation the MSCs were bred commonly, and the labeled MSCs were implanted from portal vein. CCL4 plus caudal vein transplantation group: In 3 days before MSCs transplantation, the rats were administrated with 20 g/L CCL42.5 mL/kg by gastric perfusion every day. The dose was double at the first time. Labeled MSCs were implanted from caudal vein. Caudal vein transplantation control group: Before transplantation the MSCs were bred commonly, and the labeled MSCs were implanted from caudal vein. Mixed group: On the basis of the former 4 groups, 2 rats were implanted with non-labeled MSCs; Another 2 rats fed with CCL4 for 3 days and normal feed were established, without MSCs transplantation. ③At days 3 and 7 after transplantation expression of transplanted MSCs in liver of rats of each group were examined with fluorescent quantitative PCR.MAIN OUTCOME MEASURES: ①Results of MSCs isolation, purification, in vitro amplification and phenotype identification, ②result of GFP-labeled MSCs, ③observation of growth of rats following allografting of MSCs, and ④result of quantitative identification of GFP positive DNA amount in hepatic tissues of each group.RESULTS: Totally 36 experimental SD rats were involved in the result analysis. ①Percoll gradient separating medium was applied to isolate bone marrow of rats. The obtained cells were transferred and amplified,and then mostly showed coincident shuttle shape. Cells did not express CD34 and CD45, but CD29, CD44 and CD90 of MSCs, which were noncommitted stem cells in non-differentiating status that were different from hemopoietic stem cells in bone marrow. ②The green fluorescent cells appeared 24 hours after MSCs transfection. From hour 48 to 72 the number of positive cells significantly increased, with strong intensity.The transfection efficiency was 20%-30% under high-power field, and most of the cells were with green fluorescence. But green fluorescent cells did not appear in the MSCs cells as control. ③After allografting of labeled or non-labeled MSCs of rats with different approaches, at day 1 the rats were listless with bad food appetite, less mobilization; At day 2mostly of them had normal diet and mood, but there was no significant difference in rats of each group. ④The rats in each group with the exception of mixed group had green fluorescent protein positive cells in liver at days 3 and 7. The number of green fluorescent protein positive DNA was higher in liver tissues in the CCL4 plus portal vein transplantation group and CCL4 plus caudal vein transplantation group than in the portal vein transplantation control group and caudal vein transplantation control group (P＜0.05).CONCLUSION: Duration and amount of stem cells colonizing in liver may be associated with liver injury, while not related to the implantation approach. In normal animals with uninjured liver the stem cells can colonize in liver, and the amount is associated with transplantation approach and post-transplantation duration.

Objective To compare the effects of different methods of general anesthesia on postoperative cognitive function in patients undergoing non-cardiac surgery.Methods One thousand ASA Ⅰ or Ⅱ patients,aged 18-60 years and undergoing non-cardiac surgery,were randomly divided into five groups (n=200 each):isoflurane + propofol + fentanyl group (group IPF),isoflurane + remifentanil group (group IR),sevoflurane + propofol + fentanyl group (group SPF),sevoflurane + remifentanil group (group SR),and propofol + remifentanil group (group PR).Two hundred patients receiving non-operative treatment served as control group (group C).In groups IPF and SPF,anesthesia was maintained with inhalation of 1.68％ isoflurane or 1.71％ sevoflurane,target controlled infusion (TCI) of propofol with the target plasma concentration of 2-5 μg/ml,and intermittent intravenous boluses of fentanyl.In groups IR,SR and PR,anesthesia was maintained with inhalation of 1.68％ isoflurane or 1.71 ％ sevoflurane,or TCI of propofol with the target plasma concentration of 2-5 μg/ml,and TCI of remifentanil with the target plasma concentration of 2-6 ng/ml.The patients' cognitive function was assessed with minimental state examination (MMSE) 1 day before operation,when leaving the post-anesthetic care unit (PACU),and 1 and 3 days after operation,respectively.Z score was used to identify the cognitive dysfunction as recommended by Moiler when leaving the PACU,and 1 and 3 days after operation.Results Compared with group C,the MMSE score was significantly decreased when leaving the PACU,and the incidence of cognitive dysfunction increased when leaving the PACU and 1 day after operation in the other groups (P ＜ 0.05).Compared with groups IPF,IR,SPF and PR,the incidence of cognitive dysfunction was significantly increased in group SR (P＜0.05).Conclusion General anesthesia with sevoflurane combined remifentanil exerts fewer effects on the postoperative cognitive function in patients undergoing non-cardiac surgery.

Objective To compare the effects of isoflurane or sevoflurane in combination with remifentanil anesthesia on blood amyloid beta protein (Aβ) in the elderly patients undergoing abdominal surgery.Methods Two hundred patients of both sexes,aged 65-75 yr,weighing 51-76 kg,of ASA physical status Ⅰ or Ⅱ,scheduled for elective abdominal surgery under general anesthesia,were randomly divided into 2 groups (n =100 each) using a random number table:isoflurane combined with remifentanil anesthesia group (IR group) and sevoflurane combined with remifentanil anesthesia group (SR group).Fifty healthy elderly subjects served as control group (group C).After anesthesia was induced with iv penehyclidine,sufentanil,propofol and vecuronium,the patients were endotracheally intubated and mechanically ventilated.In group IR,anesthesia was maintained with inhalation of isoflurane (end-tidal concentration 1.68 ％,in IR group) or sevoflurane (end-tidal concentration 1.71％,in SR group),and target-controlled infusion of remifentanil (target plasma concentration 2-6 ng/ml).At l day before surgery and 3 days after surgery,the patients' cognitive function was assessed using Mini-Mental State Examination (MMSE),the development of postoperative cognitive dysfunction (POCD) was recorded,and blood samples were taken for determination of serum Aβ40 and Aβ42 concentrations.Results The incidence of POCD was 5％ (in C group),56％ (in IR group) or 22％ (in SR group),and there was no significant difference among the three groups.There were no significant differences in the serum Aβ42 and Aβ40 concentrations after surgery among the three groups.Conclusion The mechanism by which sevoflurane or isoflurane in combination with remifentanil anesthesia results in POCD is not related to the levels of blood Aβ40 or Aβ42 in the elderly patients undergoing abdominal surgery.

Objective To compare the effects of methods of general anesthesia on postoperative cognitive function in patients undergoing non-cardiac surgery. Methods One thousand ASA Ⅰ or Ⅱ patients, aged 18-60 yr, undergoing non-cardiac surgery were randomly divided into 5 groups ( n = 200 each) : isoflurane + propofol + fentanyl group (group IPF); isoflurane + remifentanil group (group IR) ; sevoflurane + propofol + fentanyl group (group SPF) ; sevoflurane + remifentanil group (group SR) ; propofol + remifentanil group (group PR) . Two hundred non-operative patients served as control group (group C) . In groups IPF and SPF, anesthesia was maintained with inhalation of 1.68% isoflurane or 1.71 % sevoflurane, TCI of propofol with the target plasma concentration of 2-5 μg/ml, and intermittent iv boluses of fentanyl. In groups IR, SR and PR, anesthesia was maintained with inhalation of 1.68% isoflurane or 1.71% sevoflurane, or TCI of propofol with the target plasma concentration of 2- 5 μg/ml, and TCI of remifentanil with the target plasma concentration of 2-6 ng/ml. The patients' cognitive function was assessed using mini-mental state examination (MMSE) at 1 d before operation, while leaving postanesthesia care unit (PACU) , and at 1 and 3 d after operation. The Z score was used to identify the cognitive dysfunction as recommended by Moller while leaving PACU, and at 1 and 3 d after operation. Results Compared with group C, the MMSE score was significantly decreased while leaving PACU , and the incidence of cognitive dysfunction increased while leaving PACU and at 1 d after operation in the other groups ( P ＜ 0.05). Compared with groups IPF,IR,SPF and PR, the incidence of cognitive dysfunction was significantly increased in group SR ( P ＜ 0.05) . Conclusion General anesthesia with sevoflurane combined remifentanil exerts less effect on the postoperative cognitive function in patients undergoing non-cardiac surgery.

Objective To evaluate the relationship between sevoflurane-induced cognitive decline and α1A norepinephrine receptor (ADRA1A) in the cerebral cortex of rats.Methods Forty-eight cleangrade healthy adult Sprague-Dawley rats (24 male,24 female),weighing 220-260 g,aged 3-4 months old,were divided into 2 groups (n =24 each) using a random number table method:control group (group C) and sevoflurane group (S group).Group S inhaled 3％ sevoflurane for 5 h.Rats underwent the Barnes maze test on days 1 and 7 after anesthesia.Rats were sacrificed immediately after anesthesia and on days 1 and 7 after anesthesia,and the cerebral cortex was removed for determination of the expression of ADRA1A protein and mRNA (by Western blot or fluorescent quantitative real-time polymerase chain reaction).Results Compared with group C,the number of entering incorrect holes was significantly increased,and the latency of entering the target hole and the distance were prolonged,and the expression of ADRA1A protein and mRNA in cerebral cortex was down-regulated at each time point in group S (P＜0.05).Conclusion The mechanism of sevoflurane-induced cognitive decline is related to down-regulated expression of ADRA1A in the cerebral cortex of rats.