Objective To evaluate the usefulness of mycolic acid for identification of Mycobacterium species using SMIS. Methods One hundred and eighteen clinical Mycobacterium isolates collected from Beijing Tuberculosis and Thoracic Tumor Research Institute through whole year of 2007 were analyzed. The 118 isolates contain 25 isolates of Mycobacterium tuberculosis and 93 non tuberculosis Mycobacterium identified by PNB method. Mycolic acid analysis using SMIS is evaluated for identification of a broad range of Mycobacteria in comparison with 16S rDNA , 16-23S rDNA ITS sequencing to measure the concordance rate and agreement, and verify the concordance rate and agreement among results of mycolic acid, sequencing and PNB in identifying Mycobacterium tuberculosis and non tuberculosis Mycobacterium. Results The concordance rate between mycolic acid method analysis and DNA sequencing is 92% ( 108/118), of which concordance rate in Mycobacterium tuberculosis complex and non tuberculosis Mycobacterium are 95% (35/37) and 90% (73/81) respectively, agreement of both is great( agreement Kappa value is 0. 96). Through retrospective analysis, the concordance of results between SMIS and PNB method analysis is 90% (106/118)and agreement is well( agreement Kappa value is 0. 73 ), the concordance of results between sequencing and PNB method analysis is also 90% ( 106/118 ) and agreement is well (agreement Kappa value is 0. 74 ),despite the identification results of 11 isolates by PNB method are discordant. Conclusion Mycolic acid analysis by SMIS enables rapid identification of a broad range of clinical Mycobacterium species, which could play an important role in polyphasic identification of Mycobacterium species.

Objective To investigate the effects of hydrogen-rich saline on hippocampal mammalian target of rapamycin (mTOR)/tau pathway after operation in aged rats.Methods One hundred fifty healthy male Sprague Dawley rats, aged 18 months, weighing 400-540 g, were randomly divided into 5 groups (n=30 each) using a random number table: control group (group C);sham operation group (group S);operation group (group O);hydrogen-rich saline treatment group (group HS);normal saline group (group NS).Splenectomy was performed in group O.Hydrogen-rich saline 1 ml/100 g was injected intraperitoneally at 5 min before splenectomy in group HS.Normal saline 1 ml/100g was injected intraperitoneally at 5 min before splenectomy in group NS.Morris water maze test was performed at 1, 3 and 7 days after surgery, and the escape latency and swimming distance were recorded.After the end of the test, the rats were sacrificed, and the brains were removed for examination of the pathological changes in the hippocampal CA3 region (under light microscope), and for determination of the expression of mTOR mRNA and tau mRNA (using reverse transcriptase polymerase chain reaction) and mTOR and pS396 tau (by Western blot).Results Compared with C and S groups, the escape latency and swimming distance were significantly prolonged, and the expression of mTOR protein and mRNA, tau mRNA and pS396 tau was up-regulated in O, HS and NS groups (P＜0.05).Compared with O and NS groups, the escape latency and swimming distance were significantly shortened, and the expression of mTOR protein and mRNA, tau mRNA and pS396 tau was down-regulated in group HS (P＜0.05).The pathological changes of the hippocampal tissues were mitigated in group HS when compared with group O.Conclusion The mechanism by which hydrogen-rich saline mitigates postoperative cognitive dysfunction may be associated with inhibited activation of hippocampal mTOR/tau pathway in aged rats.

Objective To investigate the changes in the expression of hippocampal occludin and claudin-5 after operation in aged rats.Methods Eighty-one healthy male Wistar rats, aged 16-18 months, weighing 400-500 g, were randomly assigned into 3 groups (n =27 each) using a random number table: normal control group (group C), skin incision group (group I) and splenectomy group (group S).Splenectomy was performed in group S.Only skin incision and closing were performed in group I.Morris water maze test was performed before operation and on 1, 3 and 7 days after operation.The escape latency and the percentage of time spent at the target platform quadrant were recorded.Nine rats randomly selected from each group were sacrificed after the end of the test, and the hippocampi were isolated to detect the expression of occludin and claudin-5 by Western blot.Results Compared with group C, the escape latency was significantly prolonged, the percentage of time spent at the target platform quadrant was decreased, the expression of hyperphosphorylated occludin and claudin-5 was down-regulated, and the expression of hypophosphorylated occludin was up-regulated on 1 and 3 days after operation in group S (P＜0.05).Conclusion The mechanism of postoperative cognitive dysfunction is related to the downregulated expression of hippocampal hyperphosphorylated occludin and claudin-5 in aged rats.

Objective To evaluate the role of hippocampal adenosine monophosphate-activated protein kinase (AMPK) signaling pathway in reduction of postoperative cognitive dysfunction by electroacupuncture (EA) preconditioning in aged rats.Methods A total of 150 healthy male Sprague-Dawley rats,aged 18-20 months,weighing 400-500 g,were randomly divided into 5 groups (n=30 each) using a random number table:control group (group C);splenectomy group (group O);preconditioning with EA at non-acupoint group (group NEA);preconditioning with EA at Baihui acupoint group (group EA);preconditioning with EA at Baihui acupoint + AMPK inhibitor group (group EAC).EA and EAC groups received EA at Baihui acupoints with a sparse-dense wave at an intensity of 1 mA and a frequency of 2 Hz/15 Hz for 30 nin,once a day,for 5 consecutive days,and splenectomy was performed at 24 h after the end of the last stimulation.Group NEA received EA at the points 2 mm lateral to the acupoints of Baihui,and the other treatments were similar to those previously described in group EA.Compound C 20 mg/kg was injected intraperitoneally at 5 min before operation in group EAC.Morris water maze test was performed at 1,3 and 5 days after operation,and the escape latency and swimming distance were recorded.The rats were then sacrificed,and brains were removed for examination of the pathological changes in the hippocampal CA3 region and for determination of the expression of AMPK,phosphorylated AMPK (p-AMPK),nuclear factor kappa B (NF-κB),interleukin-1beta (IL-1β),and tumor necrosis factor-alpha (TNF-α) by Western blot.Results Compared with group C,the escape latency and swimming distance were significantly prolonged,and the expression of AMPK,p-AMPK,NF-κB,IL-13,and TNF-α was significantly up-regulated in O,NEA,EA and EAC groups (P＜0.05).Compared with group O,the escape latency and swimming distance were significantly shortened,and the expression of AMPK,p-AMPK,NF-κB,IL-1β and TNF-α was significantly down-regulated in EA and EAC groups (P＜0.05),and no significant change was found in the parameters mentioned above in group NEA (P＞0.05).Compared with group EA,the escape latency and swimming distance were significantly prolonged,the expression of AMPK and p-AMPK was significantly down-regulated,and the expression of NF-κB,IL-1β and TNF-α was significantly up-regulated in group EAC (P＜0.05).Conclusion Activation of hippocampal AMPK signaling pathway is involved in EA preconditioning-induced improvement in postoperative cognitive dysfunction in aged rats.

Objective To evaluate the changes in cholinergic anti-inflammatory pathway in hippocampi global in aged rats with cerebral ischemia/reperfusion (I/R ) injury .Methods One hundred and twenty male Sprague-Dawley rats , aged 18-22 months ,weighing 450-600 g ,were randomly divided into 2 groups ( n= 60 each):sham operation group (group S) and global cerebral I/R group (group I/R) .The animals were anesthetized with intraperitoneal 10% chloral hydrate 0.4 ml/100 g .Global cerebral I/R was induced by 4-vessel occlusion method described by Pulsinelli .Fifteen rats were sacrificed at 1 ,3 ,5 and 7 days of reperfusion ,and brains were removed for determination of neuronal apoptosis and expression of α7 nicotinic acetylcholine receptor (α7nAChR ) , choline acetyltransferase (ChAT ) ,tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) in the hippocampal CA1 region .The apoptosis rate was calculated .Results Compared with group S ,the apoptosis rate was increased and the expression of α7nAChR ,ChAT ,TNF-αand IL-1βwas up-regulated in group I/R ( P<0.05 or 0.01 ) . The expression of α7nAChR and ChAT was up-regulated gradually during reperfusion and peaked at 5 day of reperfusion ( P< 0.05 ) .Conclusion Global cerebral I/R injury can activate cholinergic anti-inflammatory pathway in aged rat hippocampi ,and the activation of this pathway is the endogenous mechanism of inhibition of excessive inflammatory responses in brain tissues .

Objective To evaluate the accuracy of cerebrospinal fluid (CSF) amyloid beta 42 (Aβ-42) to tau level ratio (Aβ-42/tau) in predicting postoperative cognitive dysfunction (POCD) in elderly patients.Methods Eighty ASA physical status Ⅱ or Ⅲ patients of both sexes,aged 65-85 yr,undergoing elective total hip or knee replacement under combined spinal-epidural anesthesia,were enrolled in the study.Epidural catheter was placed at T3,4 interspace,and after a catheter was successfully placed into the subarachnoid space,CSF 2 ml was obtained to measure Aβ-42 and tau levels (units:pg/ml),and the ratio between them was calculated.Neuropsychological tests were performed at 1 day before surgery and 1 week after surgery.The Z score was used to identity POCD.The cut-off value for POCD and nonPOCD patients was determined by using ROC curve.Results Thirty-two patients were diagnosed with POCD,and the incidence was 40％.The Aβ-42/tau was 2.6±0.3 and 1.7±0.4 in non-POCD and POCD groups,respectively,and the ratio was significantly lower in POCD group than in non-POCD group.The sensitivity of CSF Aβ-42/tau in predicting POCD was 91.7％,the specificity was 81.2％,and Youden index was 0.7.ROC curves revealed that the cut-off value for POCD and non-POCD patients was 2.0.Conclusion CSF Aβ-42/tau can accurately predict the occurrence of POCD in elderly patients.

Objective To evaluate the relationship between the level of AMP-activated protein kinase (AMPK) in hippocampus and cognitive dysfunction after splenectomy in aged rats.Methods Sixty-three male aged Sprague-Dawley rats,weighing 480-550 g,were randomly divided into 3 groups (n =21 each):control group (group C),anesthesia group (group A),and surgery group (group S).Morris water maze (MWM) test was performed to evaluate the spatial learning and memory ability before surgery and at 1,3 and 7 days after surgery.The escape latency and swimming distance were recorded.Seven rats were chosen after MWM test was performed at 1,3 and 7 days after surgery and sacrificed.Their hippocampi were removed for detection of the expression of AMPK and phosphorylated (p-AMPK) (by Western blot).Results Compared with the baseline,the escape latency and swimming distance were significantly prolonged at 1 and 3 days after surgery in group S (P ＜ 0.05).Compared with group C,the escape latency and swimming distance were significantly prolonged at 1 and 3 days after surgery,the expression of AMPK was up-regulated at 1,3 and 7 days after surgery,and the expression of p-AMPK was up-regulated at 1 and 3 days after surgery in group S (P ＜ 0.05),and no significant changes were found in the indices mentioned above in group A (P ＞ 0.05).Conclusion Increased AMPK level in hippocampus is the regulatory mechanism of the body adapting to the development of cognitive dysfunction after splenectomy in aged rats.

Objective To investigate the effect of mild hypothermia on glucose regulated protein 78 (GRP78) expression in hippocampus following transient cerebral ischemia-reperfusion (I/R) in aged rats.Methods One hundred and forty-four male aged Sprague-Dawley rats,weighing 450-550 g,were randomly divided into 3 groups (n=48 each): sham operation group (group S),I/R group and mild hypothermia group (group H).Global cerebral I/R was induced by 4-vessel occlusion method.Bilateral vertebral arteries were permanently occluded by cauterization,and bilateral carotid arteries were occluded for 5 min.The surface cooling was started immediately after reperfusion and maintained for 3 h.During surface cooling,the body temperature was maintained at 32-34 ℃(rectal).At 6,12,24 and 48 h of reperfusion,the expression of GRP78 was determined using immuno-histochemisty and Western blot,and the viable neurons in CA1 area were counted.Results The number of viable neurons was significantly smaller at each time point,and the expression of GRP78 was significantly higher at 6,12 and 24 h of reperfusion,while lower at 48 h of reperfusion in I/R group,and the number of viable neurons was significantly smaller,and the expression of GRP78 was significantly higher at each time point in group H than in group S (P＜ 0.05).The number of viable neurons was significantly larger at 12,24 and 48 h of reperfusion,and the expression of GRP78 was significantly higher at each time point in group H than in group I/R (P ＜ 0.05).Conclusion Mild hypothermia can further up-regulate GRP78 expression in hippocampus following transient cerebral I/R in aged rats,thus reducing endoplasmic reticulum stress and cerebral I/R injury.

Objective The aim of this prospective study was to evaluate the changes in the ovarian reverse after myomectomy based on serum anti-Mullerian Hormone (AMH) levels. Methods This is a prospective longitudinal observational study. Serum AMH levels were measured at the baseline and 2 day , 3months after myomectomy in 35 women aging from 36 to 45years.Follicle stimulate hormone(FSH) and luteal hormone(LH) were measured at the same time. 35 women of the same age with fibroid who did not undergo operation were selected as control group. Result (1)AMH level is (1.54 ± 0.95)ng/mL,(1.18 ± 0.77)ng/mL,(1.50 ± 0.58 )ng/mL at 0 day, 2 days and 3 months after operation. AMH level decreased significantly at 2 days after operation (P < 0.05) and increased gradually 3 months after operation, but showed no significant change (P > 0.05).(2) Significant differences in the serial change of AMH levels existed at each time point between myomectomy group and control group (P <0.05). No significant differences in FSH or LH levels existed at each time point. Conclusion AMH is may be superior to FSH or LH in evaluating the changing of ovarian reverse. The study suggests that myomectomy affect the ovarian function for up to 3 months post-operatively , and hemorrhage during and after operation may decrease serum AMH levels.

Objective To evaluate the efficacy and safety of administration of intranasal dexmedetomidine for preoperative sedation in the pediatric patients with congenital heart disease.Methods Sixty pediatric patients of both sexes,aged 2-5 yr,weighing 10-30 kg,of ASA physical status Ⅱ or Ⅲ (NYHA Ⅰ-Ⅲ),scheduled for elective radical operation for congenital heart disease under general anesthesia,were randomly divided into 2groups (n =30 each) using a random number table:dexmedetomidine administered intranasally group (group IN) and dexmedetomidine administered intravenously group (group Ⅳ).After admission to a room for preoperative preparation,dexmedetomidine 2μg/kg was administered intranasally in group IN and dexmedetomidine 0.5 μg/kg (in 15,ml normal saline) was injected intravenously over 20 min in group Ⅳ.After admission to the operating room at 20 min after administration,oxygen was inhaled through a face mask and induction of anesthesia was performed.The patient acceptance of modes of administration,sedation degree at 20 min after administration,score of mood state of the children when they were separated from their parents,and degree of patient acceptance of oxygen inhalation via a face mask were recorded.Bradycardia,hypotension,hypertension and hyoxemia were recorded from administration to induction of anesthesia.Results Compared with IN group,the rate of patient acceptance of modes of administration was significantly decreased,and no significant changes were found in sedation degree at 20 min after administration,score of mood state of the children when they were separated from their parents,and degree of patient acceptance of oxygen inhalation via a face mask in group Ⅳ.No adverse reactions such as bradycardia,hypotension,hypertension or hyoxemia developed from administration to induction of anesthesia in the two groups.Conclusion Dexmedetomidine 2μg/kg administered intranasally can be safely and effectively used for preoperative sedation in the pediatric patients with congenital heart disease.