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Objective To evaluate the effect of oxycodone on migration of human colon cancer cells and the role of μ and κ receptors.Methods The human colon cancer HCT116 cells at the logarithmic growth phase were seeded in 24-well or in 6-well plates at a density of 1 × 106 cells/mnl (0.5 ml/well or 2 ml/well,144 wells in total).The cells were divided into 6 groups (n=24 each) using a random number table:control group (group C),1,5 and 10 μmol/L oxycodone groups (group O1,group O2 and group O3),oxycodone plus μ receptor antagonist CTOP group (group O2+CTOP) and oxycodone plus κ receptor antagonist nor-binaltorphimine group (group O2+BNI).The cells were incubated for 24 h with oxycodone 1,5 and 10 μmol/L in O1,O2 and O3 groups,respectively.The cells were incubated for 24 h with 5 μmol/L oxycodone plus 20 μmol/L CTOP and 5 μmol/L oxycodone plus nor-binahorphimin 20 μmol/L in O2+CTOP and O2+BNI groups,respectively.The invaded and migrated cells were counted,and the levels of Ras homolog gene family member A (RhoA),Rho-associated protein kinase 1 (ROCK1),matrix metalloproteinase-2 (MMP-2) and MMP-9 were detected.Results Compared with group C,the number of invaded and migrated cells was gradually decreased,and the levels of RhoA,ROCK1,MMP-2 and MMP9 were gradually decreased in O1,O2 and O3 groups (P＜0.05),and no significant change was found in the parameters mentioned above in group O2+BNI (P＞0.05).Compared with group O2,the number of invaded and migrated cells was significantly increased,and the levels of RhoA,ROCK1,MMP-2 and MMP9 were increased in group O2 + BNI (P＜0.05),and no significant change was found in the parameters mentioned above in group O2+CTOP (P＞0.05).Conclusion Oxyc odone can inhibit the migration of human colon cancer cells,and the mechanism is totally related to inhibition of RhoA/ROCKl signaling pathway activation after activating κ receptors,but not related to μ receptors.
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Objective To compare the anesthetic efficacy of ketamine and sevoflurane for foreskin ligation in the pediatric patients.Methods A total of 120 pediatric patients,aged 2-6 yr,weighing 10-18 kg,scheduled for elective foreskin ligation,were equally and randomly divided into ketamine group (group K) and sevoflurance group (group S).In group K,atropine 0.25 mg/kg and ketamine 2 mg/kgwere injected intravenously,and foreskin ligation was performed after loss of eyelash reflex.In group S,8％ sevoflurance was inhaled using the tidal volume technique,the concentration inhaled was adjusted to 4％ after loss of eyelash reflex,and then foreskin ligation was performed.The occurrence of crying before and during anesthesia induction,induction time,emergence time,occurrence of agitation during emergence from anesthesia and duration of agitation were recorded.Results Compared with group K,the rate of crying was significantly decreased,the emergence time was shortened (P＜0.05),and no significant difference was found in the induction time,incidence of agitation during emergence from anesthesia,and duration of agitation in group S (P＞0.05).Conclusion Sevoflurance provides better anesthetic efficacy than ketamine when applied for foreskin ligation in the pediatric patients.
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Objective To evaluate the role of T?type calcium channels in up?regulation of spinal Ca2+∕calmodulin?dependent protein kinase Ⅱ ( CaMKⅡ) expression in rats with neuropathic pain. Meth?ods Forty?eight male Sprague?Dawley rats, weighing 230-270 g, in which intrathecal catheters were suc?cessfully implanted, were divided into 4 groups ( n=12 each) using a random number table: sham opera?tion group (group S), neuropathic pain group (group NP), normal saline group (group NS), and T?type calcium channel blocker mibefradil group ( group M ) . The model of neuropathic pain was established by chronic compression of the dorsal root ganglion ( DRG) . Normal saline 20μl and mibefradil 200μg ( dilu?ted to 20μl in normal saline) were injected intrathecally at 5 days after compression of the DRG in NS and M groups, respectively. Before intrathecal catheter implantation ( T1 ) , before compression of the DRG ( T2 ) , at 5 days after compression of the DRG and before intrathecal administration ( T3 ) , and at 30, 60, 120 and 240 min after intrathecal administration ( T4?7 ) , the mechanical paw withdrawal threshold ( MWT) and thermal paw withdrawal latency ( TWL) were measured. The rats were sacrificed after the last measure?ment of the pain threshold at T7 , and the lumbar enlargement segments of the spinal cord were harvested for determination of CaMKⅡ expression by Western blot. Results Compared with group S, the MWT was significantly decreased, and TWL was significantly shortened at T3?7 , and the expression of spinal CaMKⅡ was significantly up?regulated in NP and M groups (P0.05). Conclusion T?type calcium channels are opened, the intra?cellular free calcium ion concentrations are increased, and activated spinal CaMKⅡ is involved in the de?velopment of neuropathic pain in rats.
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<p><b>OBJECTIVE</b>To investigate the effect of KN93, a calmodulin-dependent protein kinase II (CaMK II) inhibitor, on SH-SY5Y cell injury induced by bupivacaine hydrochloride.</p><p><b>METHODS</b>SH-SY5Y cells exposed for 24 h to 1 mmol/L KN93, 1 mmol/L bupivacaine hydrochloride, or both were examined for morphological changes and Cav3.1 protein expressions using Western blotting. The vitality and apoptosis rate of the cells at different time points during the exposures were assessed with MTT assay and flow cytometry, respectively.</p><p><b>RESULTS</b>Bupivacaine hydrochloride exposure caused obvious cell morphologial changes, reduced cell viability, increased cell apoptosis, and enhanced Cav3.1 protein expression. All these changes were partly reversed by treatment of the cells with 1 mmol/L KN93.</p><p><b>CONCLUSIONS</b>CaMKII may play a role in bupivacaine hydrochloride-induced SH-SY5Y cells injury, which is related with upregulated Cav3.1 protein expression.</p>
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Humans , Apoptosis , Bupivacaine , Calcium Channels, T-Type , Metabolism , Calcium-Calmodulin-Dependent Protein Kinase Type 2 , Metabolism , Cell Line , Cell Survival , Up-RegulationABSTRACT
Objective To evaluate the effects of sevoflurane on invasion and migration of mouse lung cancer cells induced by hypoxia.Methods Mouse Lewis lung cancer cells were inoculated in the culture plate.After being cultured for 24 h,the cells were randomly divided into 3 groups (n=18 each) using a random number table:control group (group C),hypoxia group (group H) and hypoxia+ 2％ sevoflurane group (group HS).Cells were exposed to 95％ air-5％CO2 (2 L/min) for 4 h in group C.Cells were exposed to 94％ N2-5％CO2-1％ O2 for 4 h in group H.In group HS,cells were exposed to 2％ sevoflurane and 94％ N2 (2 L/min) for 4 h.The invasion of cells was determined by Transwell assay,and the invaded cells were counted.The migration of cells was evaluated by wound healing assay,and cell migration rates were calculated.The expression of Beclin 1 and LC3 Ⅱ protein in cells was detected by Western blot.Results Compared with group C,the number of invaded cells and cell migration rates were significantly increased,and the expression of Beclin Ⅰ and LC3 Ⅱ was up-regulated in H and HS groups.Compared with group H,the number of invaded cells and cell migration rates were significantly decreased,and the expression of Beclin 1 and LC3 Ⅱ was down-regulated in group HS.Conclusion Sevoflurane can inhibit the invasion and migration of mouse lung cancer cells induced by hypoxia,and inhibition of autophagy is involved in the mechanism.
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Objective To evaluate the effects of sevoflurane on the expression of heparanase ( HPA) and fascin in lung carcinoma cells of mice. Methods Mouse LLC cells were inoculated in the culture plate. After being cultured for 24 h, the cells were equally and randomly divided into 4 groups using a random number table: control group ( group CC) , 1% sevoflurane group ( group Sev1 ) , 2% sevoflurane group ( group Sev2 ) , and 3% sevoflurane group ( group Sev3 ) . Cells in Sev1-3 groups were exposed to 1%, 2% and 3% sevoflurane, respectively, for 4 h, while cells in group CC were not exposed to sevoflurane, and all the cells were then cultured for another 24 h in an incubator. The invasion of cells was determined by Transwell invasion assay, and the invaded cells were counted. The migration of cells was determined by wound healing assay, and cell migration rates were calculated. The expression of HPA and fascin in cells was detected by Western blot. Results Compared with group CC, the number of invaded cells and cell migration rates were gradually decreased, and the expression of HPA and fascin was gradually down?regulated with increasing concentrations of sevoflurane in Sev1-3 groups. Conclusion The mechanism through which sevoflurane inhibits the metastasis of mouse lung carcinoma cells is associated with down?regulated expression of HPA and fascin.
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Objective To evaluate the effects of dexmedetomidine on the cellular immune function of the rats with scald.Methods Seventy-two healthy male Sprague-Dawley rats,weighing 200-220 g,aged 120-150 days,were randomly divided into 3 groups (n =24 each):normal control group (group C),scald group (group S) and dexmedetomidine group (group D).Thirty percent of the total body surface was shaved and then exposed to 94 ℃ water for 12 s in S and D groups.The rats were resuscitated according to Parkland formula after scald in S and D groups,and in addition,dexmedetomidine 30 μg/kg was also intraperitoneally injected immediately after scald in D group.Before the model was established (T1) and at 12 and 24 h after scald (T2,3),blood samples from the inferior vena cava were collected for determination of T lymphocyte subsets CD3 +,CD4 + and CD8 +,NK cell,C-reactive protein (CRP),interleukin-6 (IL-6),IL-10 and tumor necrosis factor-alpha (TNF-α) level.CD4+/CD8+ was calculated.Arterial blood samples were collected for blood gas analysis.Results Compared with C,the CD3+,CD4+ and NK cell levels,CD4+/CD8+,pH value,PaCO2 and PaO2 were significantly decreased,and CD8+ levels,IL-6,IL-10,TNF-α,CRP and BE negative value were increased at T2,3 in S and D groups.Compared with group S,the CD3+,CD4+,NK cell and IL-10 levels,CD4+/CD8+,pH value,PaCO2 and PaO2 were significantly increased,and CD8+ levels,IL-6,TNF-α,CRP and BE negative value were decreased at T2,3 in group D.Conclusion Dexmedetomidine can improve the cellular immune function of the rats with scald.
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Objective To evaluate the role of C-Jun N-terminal kinase (JNK) signal transduction pathway in spinal neurotoxicity induced by lidocaine in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-260 g,were randomly divided into 6 groups (n =12 each):control group (group Ⅰ),sham operation group (group Ⅱ),JNK inhibitor group (group Ⅲ),dimethyl sulfoxide (DMSO) group (group Ⅳ),lidocaine group (group Ⅴ),and JNK inhibitor and lidocaine group (group Ⅵ).Group Ⅰ received no treatment.Intrathecal catheter was placed in the subarachnoid space in group Ⅱ.SP600125 25 μg and DMSO 20 μl were injected intrathecally in Ⅲ and Ⅳ groups,respectively.In group Ⅴ,10％ lidocaine 20 μl was intrathecally injected.SP600125 25 μg was injected intrathecally and 30 min later 10％ lidocaine 20 μl was injected intrathecally in group Ⅵ.Paw withdrawal threshold to yon Frey filament stimulation (PWT) and paw withdrawal latency to nociceptive thermal stimulation (PWL) were measured before intrathecal catheter was implanted (T0),before intrathecal administration (T1) and at 4,8 and 12 h and on 1,2,3,4,5 and 6 days after intrathecal administration (T2-10).At 24 h after intrathecal administration,4 rats were randomly chosen from each group and sacrificed.Their lumbar enlargements were removed for determination of phosphorylated JNK (p-JNK) expression (using Western blot) and neuronal apoptosis (by TUNEL).The apoptotic index was calculated.Results Compared with group Ⅰ,no significant difference was found in MWT and TWL in Ⅱ,Ⅲ groups and expression of p-JNK in Ⅱ and Ⅳ groups (P ＞ 0.05),MWT at T2-4,6-8 and TWL at T2-4,7 in group Ⅴ and MWT at T2-6 and TWL at T2-5 in group Ⅵ were significantly increased,the expression of p-JNK was down-regulated and the apoptotic index was decreased in group Ⅲ (P ＜ 0.05),and the expression of p-JNK was up-regulated and the apoptotic index was increased in Ⅴ and Ⅵ groups (P ＜ 0.05).Compared with group Ⅴ,MWT and TWL were significantly decreased,the expression of pJNK was down-regulated and the apoptotic index was decreased in group Ⅵ (P ＜ 0.05).Conclusion Activation of JNK signal transduction pathway is involved in spinal neurotoxicity induced by lidocaine in rats possibly through promoting neuronal apoptosis in the spinal cord.
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Objective To observe the expression of calcium/calmodulin dependent protein kinase Ⅱ (CaMK Ⅱ) in the spinal cord of the rats followed lidocaine hydrochloride intrathecal injection.Methods 48 male SD rats weight(230 ± 20) g,after intrathecal indwelling catheter,were randomly divided into four groups (n =12,8 rats for behavioral detection and 4 rats for western blotting):normal group (C group),sham group (S group),DMSO group (D group),10％ lidocaine group (L group).Mechanical withdrawal threshold(MWT) and thermal withdrawal latency(TWL) were detected before and after 2 h,4 h,8 h,12 h,1 d,2 d,3 d,4 d and 5 d with drug treatment.Intumescentia lumbalis of the spinal cord were collected to measure the expression of CaMK Ⅱ with western blotting after drug treatment for 12 h.Results The based MWT of the rats in C,S and D group were (11.2 ± 3.1) g,(11.8 ± 2.2) g and (11.4 ± 2.4) g respectively.There were no differences among the every time points (n=8,P＞0.05).The MWT of the rats in L group significantly increased at 2 h,4 h,8 h,12 h,1 d,2 d,3 d and 4 d after treatment with lidocaine hydrochloride,and the data were (22.0 ± 6.6) g,(22.2 ± 5.3) g,(20.5 ±5.8)g,(18.5 ±4.3)g,(16.7 ±3.2)g,(15.2 ±3.1)g,(15.5 ±3.5)g,(13.7 ±2.4)g respectively (n=8,P＜0.01).TWL had no difference among the rats in C,S,and D group(n=8,P＞0.05).The TWL of the rats in L group significantly increased at 2 h,4 h,8 h,12 h,1 d,2 d and 3 d after treatment with lidocaine hydrochloride(n =8,P＜ 0.01).The expression of CaMK Ⅱ of the rats in C group,S group,D group and L group were 0.17 ± 0.03,0.16 ± 0.03,0.19 ± 0.05,0.42 ± 0.11,and significantly upregulated in L goup (n =4,P ＜ 0.01).Conclusion Lidocaine hydrochloride intrathecal injection can increase the expression of the CaMK Ⅱ in the spinal cord of the rats.Those indicate that CaMK Ⅱ may be involved with the nerve damage induced by lidocaine hydrochloride.
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ObjectiveTo investigate the effects of sevoflurane on inhibition of invasive activity and migration of human adenocarcinoma cell line A549 by cisplatin.MethodsThe.human lung adenocarcinoma cell line A549 was obtained from Shanghai Cell Biology Institute,Chinese Academy of Sciences and cultured in RPMI 1640 culture medium containing 10％ fetal calf serum.The cells were inoculated in culture plate and cultured for 24 h and randomly divided into 4 groups:control group; 2.5 ％ sevoflurane group ; cisplatin group and cisplatin + 2.5 ％sevoflurane group.In groups sevoflurane,cisplatin and cisplatin + sevoflurane the cells were exposed to 2.5％sevoflurane or/and cisplatin 10μmol/L for 4 h respectively.The invasive activity of the cells was evaluated by Transwell chamber assay.The migration of the cells was determined by wound healing assay.The expression of MMP-2,MMP-9,Ezrin,and Fascin in the cells was detected by Western blot.ResultsBoth 2.5％ sevoflurane and cisplatin depressed invasive activity and migration of the A549 cells and down-regulated MMP-2,MMP-9,Ezrin and Fascin expression in A549 cells.The inhibitory effects of cisplatin on the A549 cells were potentiated by 2.5 ％ sevoflurane.ConclusionSevoflurane can enhance the inhibition of invasive activity and migration of human adenocarcinoma cell line A549 by cisplatin.
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ObjectiveTo investigate the role of T-type calcium channel in the spinal neurotoxicity of intrathecal (IT) lidocaine in rats.MethodsForty-eight adult male SD rats in which IT catheter was successfully implanted,weighing 230-270 g,were randomly divided into 4 groups ( n =12 each):dimethyl sulfoxide (DMSO)group (group D),lidocaine group (group L),mibefradil + lidocaine group (group M),normal saline + lidocaine group (group N).Another 12 rats served as control group (group C).DMSO and 10％ lidocaine 20μl were injected intrathecally in groups D and L respectively.After mibefradil 200 μg/10μl and normal saline 10 μl were injected intrathecally in groups M and N respectively,10％ lidocaine 20 μl was injected intrathecally in the two groups.The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured before IT injection and at 2,4,8 and 12 h and 1,2,3,4 and 5 d after IT injection (T0-9).Four rats were sacrificed at T6 in each group and their lumbar enlargements were removed for microscopic examination.ResultsCompared with group C,no significant change in MWT and TWL was found at each time point in group D,MWT was significantly increased at T1-8 and TWL was significantly prolonged at T1-7 in groups L and N,and MWT was significantly increased at T1-6 and TWL was significantly prolonged at T1-6 in group M ( P ＜ 0.05 ).Compared with groups L and N,MWT was significantly decreased at T1-4 and TWL was significantly shortened at T1-4 in group M ( P ＜ 0.05).Pathological injury was significantly reduced in group M as compared with groups L and N.ConclusionT-type calcium channel is involved in the spinal neurotoxicity of IT lidocaine in rats.
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Objective To investigate the effects of different concentrations of parecoxib on proliferation and apoptosis of colon cancer LoVo cells.Methods The colon cancer LoVo cells were inoculated in cuhure plate and cultured for 24 h.The cells were randomly divided into 4 groups (n =6 each):control group (C group) and different concentrations of parecoxib groups (P1-3 groups).The cells were incubated with parecoxib 10,40 and 160μmol/L for 24 h in P1-3 groups respectively.The rates of proliferation inhibition were measured by MTT assay.The colony formation rates were measured by colony formation assay.The apoptotic rate was determined by flow cytometry.The Survivin and caspase-3 mRNA expression in the cells was detected by RT-PCR.Results Compared with C group,the rates of proliferation inhibition and apoptotic rate were significantly increased,the colony formation rates were significantly decreased,the expression of Survivin mRNA was down-regulated,and the expression of caspase-3 mRNA was up-regulated in a concentration-dependent manner in P1-3 groups (P ＜ 0.05).Conclusion Parecoxib can inhibit the proliferation of LoVo cells and induce the apoptosis in LoVo cells in a concentration-dependent manner through down-regulating the expression of Survivin mRNA and up-regulating the expression of caspase-3 mRNA.
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Objective To investigate the effects of dexmedetomidine on perioperative cellular immune function and micro-metastasis in blood circulation in patients undergoing radical operation for colon cancer.Methods Sixty ASA Ⅰ or Ⅱ patients,aged 38-69 yr,weighing 45-67 kg,undergoing radical operation for colon cancer,were randomly divided into 2 groups (n =30 each) ∶ dexmedetomidine group (D group) and control group (C group).Anesthesia was induced with midazolam,cisatracurium and sufentanil,and maintained with propofol,remifentanil and sevoflurane.After tracheal intubation,a loading dose of dexmedetomidine 1 μg/kg was injected intravenously,followed by infusion at 0.5μg· kg-1 · h-1 until the end of operation in D group.The equal volume of normal saline was administered in C group.Venous blood samples were obtained at 5 min before induction of anesthesia (T0),1 h after beginning of operation (T1),the end of operation (T2) and 24 h after the end of operation (T3) for determination of the levels of T lymphocyte subsets (CD3+,CD4+,CD8+) and NK cells by flow cytometry.CD4+/CD8+ ratio was calculated.Cytokeratin 20 (CK20) expression in circulation was detected by RT-PCR at T0 and T3 and the positive rate was calculated.Results Compared with the baseline value at T0,the levels of CD3+ and CD4+,CD4+/CD8+ ratio and level of NK cells were significantly decreased at T2 and T3 in group C,and the levels of CD3+ and CD4+ were significantly decreased at T2 and T3 in group D (P ＜ 0.05).The levels of CD3+,CD4+ and NK cells at T2 and T3 were significantly higher and positive rate at T3 was significantly lower in group D than in group C (P ＜ 0.05).Conclusion Dexmedetomidine can improve the cellular immune function and decrease the probability of micro-metastasis in blood circulation in patients undergoing radical operation for colon cancer.
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Objective To evaluate the role of calcium/calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) in the neuronal damage induced by lidocaine.Methods SH-SY5Y cells were seeded in 96-well plates (100 μl/hole) with a density of 5 × 105/ml and randomly divided into 4 groups (n =63 each):normal culture group (C group),CaMK Ⅱ inhibitor KN93 (K group),lidocaine group (L group) and KN93 + lidocaine group (KL group).KN93 (final concentration 1 μmol/L) was added to the culture medium and the cells were then cultured for 24 h in group K.Lidocaine (final concentration 10 mmol/L) was added to the culture medium and the cells were then cultured for 24 h in group L.KN93 (final concentration 1 μmol/L) and lidocaine (final concentration 10 mmol/L) were added to the culture medium and the cells were then cultured for 24 h in group KL.The cell morphology was examined with microscope after 24 h of incubation.The viability of cells was measured by MTT assay before incubation and at 1,6,12 and 24 h of incubation.The apoptosis in the cells was assessed by flow cytometry.The apoptotic rate was calculated.Results Compared with C and K groups,the cell viability was significantly decreased and the apoptotic rate was increased in L and KL groups (P ＜ 0.05).The cell viability was significantly higher and the apoptotic rate was lower in group KL than in group L (P ＜ 0.05).There was no significant difference in the cell viability and apoptotic rate between C group and K group (P ＞ 0.05).The pathological changes were obviousin group L and significantly reduced in group KL.Conclusion CaMK Ⅱ is involved in the neuronal damage induced by lidocaine.
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Objective To evaluate the role of calcium/calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) in the up-regulation of expression of Cav3.2 T-type calcium channels in spinal cord in a rat model of neuropathic pain (NP).Methods Forty-eight male Sprague-Dawley rats,aged 3 months,weighing 220-250 g,were randomly divided into 6 groups (n =8 each)∶ sham operation group (group S),group NP,dimethyl sulfoxide group (group D) and different concentrations of a specific CaMK Ⅱ inhibitor KN93 groups (groups K1-3).NP was produced by chronic compression of dorsal root ganglion.The rats in groups D and K1-3 received a single intrathecal injection of dimethyl sulfoxide and KN93 15,30,60 nmol/L (10 μl),respectively,on 5th day after NP.Paw withdrawal threshold to von Frey filament stimulation (MWT) and paw withdrawal latency to thermal nociceptive stimulus (TWL) were measured before NP,before intrathecal injection on 5th day after NP,and at 30 and 60 min and 3,6 and 8 h after intrathecal injection on 5th day after NP (T1-7).The rats were sacrificed after the measurement of pain threshold at T7 and their lumbar enlargements were removed to detect the expression of Cav3.2 mRNA and protein using Western blot and RT-PCR.Results Compared with group S,MWT was significantly decreased,TWL was shortened and the expression of Cav3.2 mRNA and protein was up-regulated in NP,D and K1-3 groups (P ＜ 0.05).Compared with NP group,MWT was significantly increased,TWL was prolonged and the expression of Cav3.2 mRNA and protein was down-regulated in a concentration-dependent manner in K1-3 groups (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group D (P ＞ 0.05).Conclusion CaMK Ⅱ is involved in the development and maintenance of chronic NP by up-regulating the expression of Cav3.2 T-type calcium channels in rat spinal cord.
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ObjectiveTo establish a rat model of nerve damage induced by intrathecal(IT) lidocaine.MethodsFifty-five adult male SD rats weighing 200-220 g were randomly divided into 5 groups (n =11 each):group normal control (group C); group dimethyl sulfoxide (DMSO)-the solvent(group D) and groups IT 5％,10％,15％ lidocaine (groups L5.10.15 ).IT catheter was successfully implanted without complication in groups D,L5,L1o,L15.DMSO,5％,10％ and 15％ lidocaine 20 μl were injected IT in groups D,L5,L10,L15 respectively.Motor dysfunction of hindlimb was assessed and scored (0 =normal,2 =complete block) and paw withdrawal threshold to mechanical stimulation (von Frey filaments) (MWT) and paw withdrawal latency to thermal nociceptive stimulus (TWL) were measured before (baseline) and at 1,2,3,4,5,7 d after IT administration in 8 animals in each group.Three animals in each group were sacrificed at 1 d after IT administration.The lumbar segment (L4-5) was removed for microscopic examination.ResultsThere was no significant difference in motor dysfunction score,MWT and TWL among groups C,D and L5.MWT was significantly increased and TWL prolonged at 1 and 2 d after IT administration in group L10,while in group L15 motor dysfunction score was significantly increased at 1,2 d after IT administration and MWT was significantly increased and TWL prolonged at 1,2,3 d after IT administration.There was significant histologic damage to spinal cord in groups L10 and L15.Conclusion Nerve damage can be induced by IT 10％ lidocaine.
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Objective To investigate the role of T-type calcium channel in lidocaine-induced neuronal cytotoxicity . Methods SH-SYSY cell line was a gift from cell biology laboratory of our medical university. The cells were cultured in DMEM liquid culture medium at 37℃ in incubator filled with 5% CO2 , and randomly divided into 4 groups ( n = 66 each) : control group (group C)and M, L and ML groups were exposed to 5 μmol/L mibefradil (a T-type calcium channel blocker), 10 mmol/L lidocaine and 5 μmoL/L mibefradil + 10 mmol/L lidocaine for 24 h. Cell morphology was examined by electronic microscopy at 24 h of drug exposure. Cell viability (by MTT) and neuronal apoptosis (by flow cytometry) were detected immediately before and at 1, 6, 12 and 24 h of exposure to mibefradil or/and lidocaine.Results In C and M groups, the cells demonstrated dendritic protrusions, enlarged nerve processes and dense lattice. After being exposed to lidocaine for 24 h, the dendritic protrusions disappeared,the cells decreased in size, shrinked and became round; the cell viability was significantly decreased while the neuronal apoptosis increased. The lidocaine-induced changes were significantly attenuated by co-incubation with mibefradil. ConclusionT-type calcium channel is involved in lidocaine-induced neuronal cytotoxicity.
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Objective To evaluate the changes in the expression of diplopore potassium ion channel TRESK mRNA in dorsal root ganlion (DRG) in rats with neuropathic pain (NP) .Methods Thirty-two male SD rats weighing 220-250 g were randomly divided into 2 groups ( n = 16 each) : group sham operation (group S) and group NP. NP was induced by ligation and severance of left tibial and common fibular nerves according to the technique described by Decosterd. Eight rats in each group were sacrificed 1 day before and 14 day after operation and their L4,5 DRGs in the operated side were isolated for determination of TRESK mRNA expression by RT-PCR. In the remaining 8 rats in each group paw withdrawal threshold to mechanical stimuli ( MWT) and paw withdrawal latency to a thermal nociceptive stimulus (TWL) were measured at 1 day before (baseline) and 1, 3, 5, 7, 14 day after operation. Results MWT was significantly lower in group NP than in group S. The TRESK mRNA expression in L4,5 DRGs in the operated side was significantly decreased after operation as compared with the baseline before operation in group NP and was significantly lower in group NP than in group S. Conclusion The development and maintenance of NP may be closely related with down-regulation of TRESK mRNA.
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Objective To investigate the effects of different concentrations of sevoflurane on Survivin expression in human adenocarcinoma cell line A549. Methods A549 cells were obtained from Shanghai Cell Biology Medical Research Institute, Chinese Academy of Sciences and inoculated in 96 well culture plate. After being cultured for 24 h, the cells were randomly divided into 4 groups: Ⅰ , Ⅱ , Ⅲ and Ⅳ groups exposed to 95 % O2 -5 %CO2,1.7%, 3.4% and 5.1% sevoflurane respectively. A549 cells were exposed to sevoflurane for 2, 4 and 6 h respectively and then cultured for another 48 h in Ⅱ , Ⅲ and Ⅳ groups. Proliferation of A549 cells were measured by methyl thiazolyl tetrazolium (MTT) assay, and apoptosis was detected with flow cytometer at 48 h after 2, 4 and 6 h sevoflurane exposure. The expression of Survivin in A549 cells was determined by Western blot analysis at 48h after 4 h sevoflurane exposure. Results The rate of proliferation inhibition and percentage of apoptotic cells were significantly higher while the expression of Survivin was significantly lower in a concentration-dependent manner in Ⅱ , Ⅲ and Ⅳ groups as compared with group Ⅰ . Conclusion Sevoflurane can inhibit proliferation and induce apoptosis of A549 cells by inhibition of Survivin expression.
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Objective To investigate the effects of isoflurane and sevonumne on apoptosis and expression of CD44 and CD54 in human lung cancer cell line A549.Methods Human lung cancer A549 cells were obtained from Shanghai Cell Biology Medical Research Institute,Chinese Academy of Sciences,and inoculated in 24 well culture plate.After being cultured for 24 h the cells were randomly divided into 3 groups:group Ⅰ control(group C);group Ⅱ isoflurane (group Iso) and group Ⅲ sevoflurane (group Sev).A 549 cells were exposed to 1.7% isoflurane and 2.5%sevoflurane for 4 h respectively in group Iso and Sev respectively,and were then cultured for another 24 h.Apoptosis and expression of CD44 and CD54 in A549 cells were detected with flow cytometer at 0 (T0),2 h(T1) and 4 h(T2) of and 24 h after(T3) exposure to isoflurane and sevoflurane.Results The percentage of apoptotic cells wag significantly higher at T2 and T3 in group Iso than in group C.The percentage of apoptotic cells was significantly higher at T1,T2 and T3 in group Sev than in group Iso and C.The expression of CD44 and CD54 at T1,T2 and T3 was significantly decreased as compared with the baseline at T0 in group Iso and Sev and was significantly lower in group Iso and Sev than in group C.Conclusion Isoflurane and sevoflurane can induce apoptesis of human lung cancer cell line A549, and sevoflurane is more effective. Isoflurane and sevoflurane can inhibit the expression of CD44 and CD54 of human lung cancer cell line A549.