Objective To evaluate the role of extracellular signal-regulated kinase-cyclic AMP response element binding protein(ERK-CREB) signal pathway in glucocorticoid receptors-mediated chronic morphine tolerance in rats.Methods Male SD rats aged 2 months weighing 280-320 g were used in this study.A catheter was placed in subarachnoid space via foramen magnum according to Yaksh.Thirty-six rats in which intrathecal (IT) catheters were successfully implanted were randomly divided into 6 groups ( n =6 each):control group ( group C),chronic morphine tolerance group (group M),morphine + dexamethasone group (group MD),morphine + RU38486 group (group MR),dexamethasone group (group D),RU38486 group (group R).Normal saline 10 μl,morphine 10 μg,morphine 10 μg + dexamethasone 4 μg,morphine 10 μg + RU38486 2 μg,dexamethasone 4μg,RU38486 2 μg was administered IT twice a day(8:00 and 20:00)for 6 consecutive days in groups C,M,MD,MR,D,R respectively.Tail flick latency (TFL) was measured at 1 d before IT drug administration(baseline)and at 30 min after first IT drug administration during 1,3,5 d and at 1 d after last IT drug administration (T1～4).Maximum analgesic effect (MPE) was calculated.The animals were sacrificed after last TEL measurement.The L3～5 segment of the spinal cord was isolated for determination of the expression of phosphorylated ERK(pERK)and phosphorylated CREB(pCREB) by immunofluorescence staining.Results MPE was significantly T1 lower at T3,4 than at T1 in groups M and MD.Compared with group C,MPE was significantly increased,the expression of pERK and pCREB up-regulated in group M,but no significant change was found in the parameters mentioned above in groups R and D.Compared with group M,MPE was significantly increased,the expression of pERK and pCREB up-regulated in group MR,and MPE was significantly increased,the expression of pERK and pCREB down-regulated in group MD.Conclusion The mechanism by which glucocorticoid receptors-mediated chronic morphine tolerance may be associated with the inhibition of ERK-CREB pathway.

ObjectiveTo evaluate the role of the spinal cord proteasome in chronic morphine tolerance in rats.MethodsTwenty-four healthy male SD rats in which intrathecal catheters were successfully placed without complications were randomized into 4 groups ( n =6 each):normal saline group ( group NS),chronic morphine tolerance group (group M),chronic morphine tolerance + proteasome inhibitor MG-132 group (group M + MG) and MG-132 group (group MG).Normal saline 10 μl,morphine 10 μg,morphine 10 μg+ MG-132 2.5 μg and MG-132 2.5 μg were injected intrathecally twice a day for 7 consecutive days in groups NS,M,M + MG and MG respectively.Tail flick latency was measured on day 1 before intrathecal injection and on day 1,3,5 and 7 of intrathecal injection to calculate the percentage of maximum possible antinociceptive effect (MPAE).After the last intrathecal injection,5 rats were sacrificed,and L3-5 spinal cord was removed for determination of the expression of glutamate-aspartate transporter (GLAST) and excitatory amino acids carrier 1 (EAAC1)by Western blot.Results MPAE was gradually decreased during the intrathecal injection in groups M and M + MG( P ＜ 0.05).Compared with group NS,MPAE was gradually incresed during the intrathecal injection in groups M and M + MG,and the expression of GLAST and EAAC1 in the spinal cord was significantly down-regulated in group M (P ＜ 0.05).There was no significant difference in the parameters mentioned above between group MG and group NS ( P ＞0.05 ).Compared with group M,MPAE was significantly increased and the expression of GLAST and EAAC1 in the spinal cord was significantlyup-regulatedingroupM+MG(P＜0.05or0.01).ConclusionSpinal cord proteasome is involved in the development of chronic morphine tolerance in rats.

The use of genetic approaches to probe relative genes that control sensitivity to volatile anesthetics in intact model has recently emerged as the powerful tools and strategies in dissecting mechanisms of anesthesia. Multiple model organisms such as yeast, nematodes, fruitflies and mammals are currently being exploited, and a number of sensitive genes have been screened, with some of them being cloned, located, and function identified. The emerging technologies are likely to provide further great advances for elucidating the specific anesthetic molecular sites.

G1.(3)Among 5 groups,the highest incidences of nausea,vomiting and pruritis were observed in group G1 for about 24% 32%,but with no statistical difference compared with other groups.Conclusion: With equal doses of bupivacaine and fentanyl mixed,the concentration/volume ratios may affect the analgesic effects of postoperative epidural analgesia in patients with hepatectomy.

Objective To compare HASTE-MRCP single-slice acquisition technique with ordinary MR cholangiopancreatography (OMRCP) and to assess the clinical application value of HASTE-MRCP single-slice acquisition technique.Methods 48 cases with biliary obstructive diseases and 30 cases without obstructive jaundice were studied with a 3. 0T super conductive unit(Magnetom Trio;Siemens,Germany) that used a body coil. Two acquisition techniques of MRCP were used for all patients . The accuracy of the two kinds of technique were compared. Results OMRCP , single-slice HASTE-MRCP had the same sensitivity and specificity in showing the site and feature of the diseases , the proximal dilated biliarys and distant normal cholangiopancreatic ducts .The diameter of dilated bile ducts at the same level was compared and no significant difference in them. Single-slice HASTE-MRCP had a high spatial resolution ,and shot aquisition time ,but more vessel artifact. OMRCP had high density resoultion ,but more motion artifacts.Conclusion The single-slice acquisition of HASTE-MRCP is an excellent technique in the diagnosis of biliary tract diseases.

Objective To investigate the role of spinal glucocorticoid receptors (GR) in phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signal pathway in rats with morphine tolerance.Methods Forty healthy male SD rats aged 8-10 weeks weighing 300-350 g in which intrathecal (IT) catheters were successfully implanted without complication were randomly divided into 4 groups (n =10 each):control group(group C) received IT injection of normal saline 10 μl twice a day for 7 consecutive days; morphine tolerance group(group M) received IT injection of morphine 10 μg twice a day for 7 consecutive days; dexamethasone (a GR agonist) group( group DEX)received IT injection of dexamethasone 4 μg 30 min before IT injection of morphine,twice a day for 7 consecutive days;RU38486(a GR blocker)group (group R) received IT injection of RU38486 2 μg 30 min before IT injection of morphine,twice a day for 7 consecutive days.Tail-flick test was measured once a day after first IT administration and 1 d after the end of IT administration,and the percentage of maximum possible antinociceptive effect (MPAE)was caculated.After the last measurem of tail-flick test,the spinal dorsal horns were removed for determination of PI3K,Caspase-3 expression and Akt activity.Results Morphine tolerance developed in groups M,DEX and R,but did not develop in group C.Compared with group C,Akt activity was decreased,PI3K expression was downregulated and Caspase-3 expression was up-regulated in group M (P ＜ 0.05).Compared with group M,MPAE and Akt activity were decreased,PI3K expression was down-regulated and Caspase-3 expression was up-regulated in group DEX,and MPAE and Akt activity were inecreased,PI3K expression was up-regulated and Caspase-3 expression was down-regulated in group R (P ＜ 0.05).Conclusion Spinal cord GR is involved in morphine tolerance by inhibiting PI3K/Akt signal pathway.

ObjectiveTo evaluate the effect of hydrogen-rich saline given during reperfusion on global cerebral ischeraia-reperfusion (I/R) injury in rats.Methods Seventy-two adult male SD rats,aged 2.0-2.5 months,weighing 260-300 g,were randomly divided into 3 groups (n ＝ 24 each):sham operation group (group S),group I/R and hydrogen-rich saline group (group H).In groups I/R and H cerebral I/R was induced by occlusion of 4 vessels( cauterization of bilateral carotid arteries and 15 min occlusion of bilateral common carotid arteries).In group H intraperitoneal 0.6 mmol/L hydrogen-rich saline 5 ml/kg was injected at 6 h of reperfusion,while equal volume of normal saline was injected instead of hydrogen-rich saline.Eighteen rats of each group were sacririced at 24 h of reperfusion,and then the hippocampi were removed for determination of malondialdehyde (MDA),tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6)contents,and nuclear factor-κB (NF-κB) activity and activated caspase-3 expression.Another six rats of each group were sacrificed at 72 h of reperfusion,and then brain tissues were removed for microscopic examination and counting the number of uninjured pyramidal cells in hippocampal CA1 region.ResultsCompared with S group,the contents of MDA,TNF-α,IL-6 and NF-κB activity were significantly increased,activated caspase-3 expression was significantly up-regulated,uninjured pyramidal cells in hippocampal CA1 region were significantly decreased in I/R group( P < 0.05).Hydrogen-rich saline given during reperfusion attenuated the above-mentioned I/R-induced changes( P < 0.05 ).The histologic damage of the hippocampal CA1 region was significantly slighter in group H than group I/R.ConclusionHydrogen-rich saline given during reperfusion can reduce global cerebral I/R injury in rats through inhibition of lipid peroxidation,inflammatory response and apoptosis.

In order to understand the resistance against common antibiotics in clinic and macrolide antibiotic-resistant mechanism of Streptococcus pneumoniae (S.pneumoniae) isolates in Zhejiang area,both K-B slip method and E-test were applied to determine the sensitivity of 138 S.pneumoniae isolates to nine antibiotics,and the ermB and mefE genes in those isolates which associated with macrolide antibiotics-resistance closely were detected by PCR.Subsequently,correlation among ermB and mefE genes and the erythromycin resistance were analyzed.For these 138 S.pneumoniae isolates,93.5% (129/138) of the strains were resistant to erythromycin,but only 2.9%～4.3% strains were resistant to cefotaxim,cefuroxime,amoxicillin and levofloxacin.The positive rate of ermB gene in the isolates (91.3%,126/138) was significantly higher than that of mefE gene (33.3%,46/138) (P<0.05).Both of these two genes existed in 27.5 % (38/138) of the strains and all of the strains without ermB and mefE genes were sensitive to erythromycin.The erythromycin resistance rate (62.5%) of mefE gene positive strains was remarkably lower than that of the mefE&ermB gene positive strains (100%) and the ermB gene positive strains (97.7%) (P<0.05).All the data mentioned above demonstrated that erythromycin is not an appropriate antibiotic to treat the infectious diseases caused by S.pneumoniae.Moreover,ermB is the predominant erythromycin resistance gene in S.pneumoniae isolates and ermB gene could inspire stronger erythromycin resistance than mefE gene.

OBJECTIVE:To evaluate therapeutic efficacy and safety of ubenimex combined with chemotherapy in the treatment of malignant tumor,and to provide evidence-based reference in clinic.METHODS:Retrieved from Central,PubMed,CJFD,VIP and Wanfang database,randomized controlled trials (RCTs) about ubenimex combined with chemotherapy (trial group) vs.single chemotherapy (control group) in the treatment of malignant tumor were collected.The quality of studies were evaluated by bias risk evaluation criteria of Cochrane system evaluator manual 5.1.0 after screening literatures and extracting data.Meta-analysis was performed by using Rev Man 5.3 statistical software.RESULTS:A total of 12 RCTs were included,involving 762 patients.The resuits of Meta-analysis showed that:shorter-term response rate [RR=1.24,95 ％ CI (1.08,1.43),P=0.002] and the improvement rate of life quality Karnofsky score [RR=1.69,95％ CI(1.46,1.95),P＜0.001] in trial group were significantly higher than control group;the incidence of gastrointestinal toxicity [RR=0.74,95％CI(0.57,0.94),P=0.02] and leucocyte suppression rate[Ⅰ °-Ⅳ°(＜3 months):RR=0.54,95％CI(0.37,0.79),P=0.002;Ⅲ°-Ⅳ°:RR=0.44,95％CI(0.29,0.68),P＜0.001] were significantly lower than control group,with statistical significance.CONCLUSIONS:Ubenimex combined with chemotherapy can improve malignant tumor,shorter-term efficacy and life quality,and reduce gastrointestinal and marrow toxicity.

Objective To discuss the adjustment effects of kidney reinforcing medicine on growth and development and the content of choline acetyl transferase (CHAc) in rats with kidney-deficiency constitution. Methods The method of cats scare rats was used to build composite offspring rat models with deficiency and acquired dystrophy, and then the models were divided into model group, Zuogui Pill group and Yougui Pill group. The rats in blank group came from normal pregnant rats. Baby rats were scared and received gavage at the same time. All administration groups received suspension of Zuigui Pills or Yougui Pills. Blank group and model group received the same amount of normal saline, once a day, for 3 months. Growth and development of rats were observed. Weight and food utilization of rats aged 5–8 weeks in each group were recorded. The content of brain CHAc was detected by ELISA. Results When offspring were 5–8 weeks old, weights of rats in the model group were lower than blank group (P<0.01); while weights of rats in Zuogui Pill group and Yougui Pill group increased significantly (P<0.05) and the food utilization was positively correlated with weight; the CHAc content in the model group decreased significantly compared with blank group (P<0.05, P<0.01); while the CHAc content in Zuogui Pill group and Yougui Pill group increased significantly compared with the model group (P<0.01). Conclusion Kidney reinforcing medicine can improve backward growth and development of rats with kidney-deficiency constitution and adjust CHAc in the brain, so as to promote the learning and memory ability of rats with kidney-deficiency constitution.