ObjectiveTo study the suppressive role of emodin on the growth and its effect on the proliferation cycle and apoptosis of human bladder cancer cell line BIU87.MethodsThe effect of different concentration of emodin at different time point on cell proliferation of BIU87 were measured with methylthiazole (MTT) chromatometry, the cell proliferation cycle were detected with flow cytometry, expressions of bc1-2 and caspase-3 were detected by SP of immunohistochemistry.ResultsWithin a certain range, the higher concentration of emodin (10 ～ 80 μg/ml) and the longer action time are positive related the more significant inhibition of tumor cell growth and the higher apoptosis rate [(9.84 ± 1.13)％, (18.32 ±2.14)％ ,(29.73+1.42)％ ,(42.13 +2.36)％ ,respectively].Compared with control group [(2.01 ±0.92)％], the differences were statistically significant(F =531.85, P ＜0.01).Emodin could inhibit the proliferation of human bladder cancer cell BIU87 by blocking BIU87 cell in G0/G1 stage, thus cut down cell proportion in stage of S [(33.27 +1.26)％ ,(29.17 ±1.39)％, (16.94 ±0.86)％ ,(10.85 ± 1.47)％,respectively], compared with the control group [(35.45 ± 0.38) ％], the differences were statistically significant(F =524.64, P ＜0.01).After 48 h of emodin treatment, the bc1-2 expression(Grayscale values:122.65 + 2.12,131.37 ± 1.62,134.81 ± 1.36,145.55 ± 2.01, respectively) was decreased and the caspase-3 expression(Grayscale values : 135.26 + 1.41,130.22 ± 1.74,126.11 ± 1.77,118.36 + 1.53, respectively) was increased in a dose dependent manner.Compared with control group (Grayseale values:108.42 + 3.73,149.35 ± 1.82, respectively), the differences were statistically significant (F = 216.23,224.83, P ＜0.01).ConclusionsEmodin could significantly inhibit the growth and induce apoptosis of BIU87 cells in vitro, which may be through down regulation of bc1-2, and up regulation of caspase-3, and blocking BIU87 cell in G0/G1 stage.

Objective To investigate the inhibitory effect of emodin on hetertransplanted human bladder cancer in nude mice and explore its mechanism.Methods Heterotransplanted models of human bladder cancer cell line BIU87 cells in nude mice were established.The mice were randomly divided into 4 groups during the experiment:blank control group,Z-VAD-FMK group,emodin group and emodin combined with Z-VAD-FMK group.The growth of tumors was observed and the growth curve was mapped.The nude mice were sacrificed 4 weeks later,the tumors were isolated and weighed.The pathological changes of tumor were observed after HE staining,the cells apoptosis were detected with flow cytometry,and the expression of AIF and Endo G were examined by reverse transcription PCR (RT-PCR) and Western blot.Results The tumor growth rate in emodin group was lower than that in the other three groups.The tumor quality in emodin group [(0.41 ± 0.05 ) g] and emodin combined with Z-VAD-FMK group [( 0.69 ±0.07)g]were lighter than that in the other two groups[(1.08 ±0.13,1.04 ±0.09)g,],and the differences were statistically significant( F =90.56,27.49,P ＜0.01 ).The quality difference in emodin group and emodin combined with Z-VAD-FMK group was statistically significant ( t =10.01,P ＜ 0.01 ).The apoptosis rate in emodin group [(42.71 ±2.69)％]was significantly higher than that in emodin combined with Z-VAD-FMK group[(34.38 ± 1.73)％] ( t =6.38,P ＜0.01 ).The expression of AIF and Endo G in emodin combined with Z-VAD-FMK group was significantly increased than other groups [( 1.65 ±0.12)vs(1.24±0.08),(0.51 ±0.07),(0.48 ±0.04);(2.12 ±0.16)vs(1.75 ±0.13),(0.57 ±0.06),(0.59±0.07);(2.42±0.13)vs(1.73 ±0.11),(0.78 ±0.07),(0.75 ±0.08);(3.13 ±0.25)vs(2.15± 0.18 ),(0.85 ± 0.09 ),(0.81 ± 0.14 )],and the differences were significant ( F =303.22,319.32,409.38,258.53,P ＜ 0.01 ).Conclusions Emodin could significantly inhibit the growth of hetertransplanted human bladder cancer in nude mice.The mechanism might be partly due to the expression increase of AIF and Endo G in bladder cancer cells,which might induce apoptosis through Caspase-independent pathway.

Objective To investigate the effects of different doses of dexmedetomidine pretreat-ment on cardiac toxicity of bupivacaine in rats.Methods Forty eight adult male SD rats weighing 250-300 g were randomly divided into 4 groups (n=12):saline control group (group C),dexmedetomi-dine 5 μg/kg group(group D5),dexmedetomidine 10 μg/kg group(group D10)and dexmedetomidine 1 5 μg/kg group(group D1 5 ).A Ⅱ-lead electrocardiogram(ECG)was continuously monitored,the femoral artery was cannulated for direct measurement of MAP and the femoral vein was cannulated for infusion of drugs.Groups D5,D10 and D1 5 were received infusion of dexmedetomidine 5,10 and 1 5μg/kg respectively 1 5 minutes before administration of bupivacaine,while the equal volume of saline was given in group C,then all rats received infusion 0.75% bupivacaine at the rate of 2 mg·kg-1· min-1 until asystole occurred.The doses of bupivacaine and the times of bupivacaine-induced convul-sions,arrhythmia and asystole were recorded respectively,and the myocardial concentration of bupiv-acaine was observed.Results Compared with group C,the doses of bupivacaine and the times of bupivacaine-induced convulsions,arrhythmia and asystole were all increased in groups D5,D10 and D1 5 (P <0.05).Compared with group D5,the above parameters were increased in groups D10 and D1 5 (P <0.05 ).There was no statistical significance of the above parameters between groups D10 and group D1 5.Conclusion Dexmedetomidine pretreatment can raise the threshold toxic dose of bupi-vacaine,delay the time of occurrence of cardiotoxicity of bupivacaine,so that to prevent the cardiac toxicities of bupivacaine in rats,and it produces a dose-dependent protective effect within a certain dose range.

Objective To study the effects of emodin on apoptosis and mRNA and protein of apoptosis inducing factor (AIF) and Endonuclease G (Endo G) in human bladder cancer cells BIU87,and to investigate the anticancer mechanism of emodin. Methods The BIU87 cells were divided into 4 groups,control group,Z-VAD-FMK group,emodin group,emodin combined with Z-VAD-FMK group.The effects of different concentrations of emodin at different action time on cells proliferation of BIU87 in vitro culture were measured by methylthiazole (MTT) chromatometry,the cells apoptosis were detected by flow cytometry,and expression of AIF and Endo G were examined by reverse transcription PCR (RT-PCR) and Western blot.Results MTT assay demonstrated that the higher concentration of emodin and the longer action time,the more significant inhibition of tumor cell growth.Based on the IC50 value,80 μmol/L and 72 h of emodin intervention were selected as an intervention condition.The apoptosis rate in emodin group (44.57 ± 1.52 ) ％was significantly higher than that in emodin combined with Z-VAD-FMK group (35.58 ± 1.61 ) ％ ( P ＜0.01).RT-PCR and Western blot showed that the mRNA and protein of AIF in emodin combined with Z-VAD-FMK group,emodin group,control group,Z-VAD-FMK group were ( 1.74 ± 0.11 ) and (2.59 ±0.13),(1.36±0.08) and (1.89±0.14),(0.37 ±0.02) and (0.53±0.11),(0.42 ±0.06) and (0.44 ± 0.07),respectively.There were significant differences between emodin group and the other groups (P ＜0.01 ).The mRNA and protein of Endo G in emodin combined with Z-VAD-FMK group,emodin group,control group,Z-VAD-FMK group were (2.28±0.15) and (3.31 ±0.36),(1.85 ±0.13) and (2.15 ±0.27),(0.53 ±0.07) and (0.71 ±0.16),(0.61 ±0.04) and (0.67 ±0.22),respectively.The differences were significant between emodin group and the other groups ( P ＜ 0.01 ). Coneltusion Emodin can upgrade the expression of AIF and Endo G in bladder cancer cells BIU87,which can induce apoptosis through Caspase-independent pathway.

Objective To evaluate the efficacy of oxycodone for patient-controlled intravenous analgesia (PCIA) after hip arthroplasty in elderly patients.Methods Sixty patients of both sexes,aged 65-85yr,weighing 42-89 kg,of American Society of Anesthesiologists physical status Ⅰ or]Ⅱ,scheduled for elective unilateral hip arthroplasty under general anesthesia,were divided into 2 groups (n =30 each) using a random number table:morphine group (group M) and oxycodone group (group O).Parecoxib sodium 40 mg was injected intravenously at 30 min before induction of anesthesia,followed by induction and maintenance of anesthesia.PCIA pump was connected at the beginning of skin closure.PCIA solution contained morphine 0.6 mg/kg and tropisetron 10 mg diluted to 100 ml in normal saline in group M and oxycodone 0.6 mg/kg and tropisetron 10 mg diluted to 100 ml in normal saline in group O.The PCA pump was set up with a background infusion at 2 ml/h,a 0.5 ml bolus dose and a 15 min lockout interval in both groups.Visual analogue scale score was maintained ≤ 3,and postoperative analgesia lasted until 48 h after operation.When analogue scale score ≥ 4,pethidine 50 mg/kg was injected muscularly as rescue analgesic.The requirement for rescue analgesic and occurrence of adverse effects were recorded.Results Ten percent patients required rescue analgesics in group M,and no patients required rescue analgesics in group O.Compared with group M,the requirement for rescue analgesics and incidence of nausea,vomiting and pruritus were significantly decreased in group O (P＜0.05).Conclusion Oxycodone provides reliable efficacy for PCIA after hip arthroplasty in elderly patients with fewer adverse effects,indicating that oxycodone produces good analgesic efficacy for severe somatalgia.

Objective To evaluate the effect of stellate ganglion block (SGB) on postoperative synaptic structure in hippocampal CA3 region in aged rats.Methods Seventy-two male Sprague-Dawley rats,aged 20-22 months,weighing 550-650 g,were randomly divided into 3 groups (n =24 each) using a random number table:control group (group C),operation group (group O) and SGB + operation group (group SGB).Group SGB received right SGB with 0.25％ bupivacaine 0.15 ml.Groups O and SGB underwent 30 min of exploratory laparotomy starting from 15 min after the end of administration.Y-maze test was performed on 1 day after operation in 6 rats chosen from each group for assessment of cognitive function.The frequency of standard training and standard time were recorded.Six rats were chosen from each group on 1,3 and 7 days after operation and sacrificed and the hippocampal CA3 region was isolated for microscopic examination and for measurement of synaptic structure.Results Compared with group C,the standard time was significantly prolonged,and the frequency of standard training was increased in groups O and SGB,the width of synaptic cleft was increased,the thickness of post-synaptic density was decreased,the length of active zones was shortened,and the curvature of the synaptic interface was decreased on 1,3 and 7 days after operation in group O (P ＜ 0.05),and no significant changes were found in each synaptic structure parameter in group SGB (P ＞ 0.05).Compared with group O,the standard time was significantly shortened,the frequency of standard training was decreased,the width of synaptic cleft was decreased,the thickness of the post-synaptic density was increased,the length of active zones was prolonged,and the curvature of the synaptic interface was increased on 1,3 and 7 days after operation in group SGB (P ＜ 0.05).Conclusion The mechanism by which SGB improves the postoperative cognitive dysfunction in aged rats may be related to inhibition of changes of synaptic structure in hippocampal CA3 region.

Objective To investigate the effects of Saikosaponin A (SSA)on cognitive function and cAMP/CREB signaling pathway and expression of BDNF in mice after traumatic brain injury. Methods Sixty SD male mice were randomized into three groups:shame operation group (group S, n =20),trauma group (group T,n =20)and SSA treatment group (group A,n =20).Mice received an administration of SSA 5 mg/kg (group A)or equal volume saline (group S,group T)immediately and once daily for 5 consecutive days after trauma.The cognitive function was detected by Morris wa-ter maze test on day 1,3,7 and 14 after trauma.The hippocampal tissues were harvested after be-havioral tests and homogenized for measuring the levels of brain derived neurophic factor (BDNF)and cyclic AMP (cAMP)by ELISA as well as the levels of cAMP-response element binding protein (CREB)and phosphorylation-cAMP-response element binding protein (pCREB)by western bolt. Results Compared with group S,the escape latency and swimming distance were significantly pro-longed in group T on day 1,3,7 and 14 and group A on day 1,3 after trauma (P <0.05 );while compared with group T,they were significantly shorter in group A on day 7,14 after trauma (P <0.05).Compared with group S,the levels of BDNF,cAMP,CREB and pCREB were significantly de-creased in group T(P < 0.05 ).Compared with group T,the levels of BDNF,cAMP,CREB and pCREB were significantly increased in group A (P <0.05).Conclusion SSA can significantly improve cognitive dysfunction in mice after traumatic brain injury,and the mechanism may be related to the activation of cAMP/CREB signaling pathway and up-regulation of BDNF.

Objective To investigate the effects and mechanism of pre-treatment with an Nrf2 inducer dh404 on renal ischemia reperfusion injury in rats.Methods Thirty SD rats were divided into three groups using completely randomized digital table,dh404 was dissolved in sesame oil and was given orally 1.5 mg/kg the night before procedures and 5 hours before procedures.Rats in group Sham received no treatment of ischemic reperfusion.In group IR and group dh404,the renal ischemia reper-fusion (IR)model was established,24 hours after IR,the levels of serum creatininc (Cr)and urea ni-trogen (BUN),the activities of superoxide dismutase (SOD)and the content of malonaldehyde (MDA)in serum were measured,and hematoxylin eosin(HE)staining observe the changes in renal structure,the levels of γ-glutamate-cysteine ligase catalytic (GCLC)and modifier (GCLM)subunit, the expression of NF-κB,COX-2 and eNOS were measured.Results Compared with group Sham,the values of Cr,BUN in group IR and group dh404 were significantly higher (P <0.05).Compared to the group IR,the group dh404 Cr,BUN values significantly decreased after reperfusion for 24 h(P <0.05 ).Compared to group Sham,group IR SOD activity decreased,while the value of MDA increased(P <0.05 ).Compared to group IR,group dh404 had much higher SOD activity,while the value of MDA significantly decreased.Observed with optical microscopy,compared to group Sham, the renal tubular injury of group IR was obvious.Compared to group IR,group dh404 significantly reduced tubular injury.Compared to group IR,the levels of GCLC and modifier GCLM subunit were higher,while there were no significant differences of levels among NF-κB,COX-2 and eNOS. Conclusion Pre-treatment with an Nrf2 inducer dh404 can protect the kidney from IRI through possi-bly reducing IRI kidney oxidative stress.

Objective To evaluate the effect of stellate ganglion block (SGB) on cellular immune function in diabetic rats.Methods Healthy male Sprague-Dawley rats,aged 3 months,weighing 240-280 g,were used in this study.Diabetes mellitus was induced by intraperitoneal 1％ streptozotocin 60 mg/kg and confirmed by blood glucose ≥ 16.7 mmol/L 3 days later.Forty-eight rats with diabetes mellitus were randomly divided into 2 groups (n=24 each) using a random number table:diabetes mellitus group (group DM) and group SGB.Another 24 healthy rats,aged 3 months,were selected and served as control group (group C).At 1 week after successful establishment of the model,unilateral transection of cervical sympathetic trunk (TCST) was performed in group SGB,while the right cervical sympathetic trunk was only exposed in C and DM groups.Before TCST (T0) and on 1,3,7 days after TCST (T1-3),6 rats were randomly selected from each group,and blood samples were collected from the inferior vena cava for determination of the blood glucose,plasma norepinephrine (NE) concentrations (by enzyme-linked immunosorbent assay),and levels of T lymphocyte subsets CD3+,CD4+ and CD8+ in whole blood (using FACSCalibur flow cytometer).C D4+/CD8+ratio was calculated.The rats were weighed before sacrifice,and the rats were sacrificed to obtain the thymus which was weighed.The thymus index (thymus weight/body weight) was calculated.Results Compared with group C,the blood glucose was significantly increased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly decreased at T0-3 (P＜0.05),and no significant change was found in CD8+ levels in DM and SGB groups (P＞0.05),the plasma NE concentrations were significantly decreased at T1-3 in group SGB (P＜0.05),and no significant change was found in plasma NE concentrations in group DM (P＞0.05).Compared with group DM,the blood glucose and plasma NE concentrations were significantly decreased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly increased at T1-3 (P＜0.05),and no significant change was found in CD8+ levels in group SGB (P＞0.05).Conclusion SGB can improve the cellular immune function in diabetic rats.