Aim To investigate the role of T type calcium channel of spinal cord and supraspinal on the pain threthold of the rats following chronic constriction injury(CCI) of the sciatic nerve.Methods Intrathecal and lateral ventricle injection were employed in this study.With Von Frey hair and radiant thermal stimulator,we measured the mechanical withdrawal threshold(MWT) and thermal withdrawal latency(TWL)of the rats after injected mibefradil.Results The rats of CCI group formed steady mechanical and heat hyperaglsia from the third day after operation to the end of this study.Administered intrathecally mibefradil 50,100,200 ?g can increase the CCI rats MWT and TWL.However,mibefradil administered lateral ventricle can reduce the CCI rats MWT and TWL.Conclusion Blocking T type calcium channel of spinal can inhibit mechanical and heat hyperalgesia of the CCI rats,However,bloking the T type calcium channel of supraspinal can enhance the mechanical and heat hyperalgesia of the CCI rats.

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.

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.

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.

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.

Objective To investigate the effect of intrathecal(IT)mibefradil on the mechanical and thermal hyperalgesia following chronic compression of dorsal mot ganglion(CCD)in rats,trying to elucidate the role of T-type calcium channels in the nociceptive signal transmission at spinal level.Methods Forty-eight male SD rats weighing 230-250 g were randomly divided into 6 groups(n=8 each):group Ⅰ sham operation;group Ⅱ CCD;group Ⅲ CCD+normal saline(N.S.)and group Ⅳ,Ⅴ,Ⅵ CCD+mibefradil 50 ?g(Ⅳ),100 ?g (Ⅴ),200 ?g(Ⅵ).The animals were anesthetized with intraperitoneal(IP)1% pentobarbital 40 mg?kg~(-1). Intrathecal catheter was placed according to the technique described by Yaksh et al.Five days after IT catheter placement L_4 and I_5 dorsal root ganglion(DRG)compression(CCD)was performed in group Ⅱ-Ⅵ.In sham operation group(Ⅰ)only L_(4-5) transverse processes and intervertebral foramina were exposed but DRGs were not compressed.In group Ⅳ,Ⅴ,and Ⅵ 5 days after CCD model was established a bolus of mibefradil 50,100 and 200 ?g in 10 ?l NS was given IT.In group Ⅲ NS 10 ?l was given IT instead of mibefradil.Mechanical withdrawal threshold(MWT)using Von Frey filament and thermal withdrawal latency(TWL)using radiant heat applied to the plantar surface were measured before CCD(baseline)and 1,3,5,7,14 and 21 days after CCD in group Ⅰ and Ⅱ and before and 30,60,120,240 and 480 min after IT mibefradil in group Ⅲ-Ⅵ.Results The animals in group Ⅱ(CCD group)developed mechanical and thermal hyperalgesia from the 1st day after operation to the end of the experiment.IT mibefradil 50,100 and 200 ?g can all attenuate both mechanical and thermal hyperalgesia induced by CCD.Conclusion Spinal T-type calcium channel may play an important role in the pathogenesis of neuropathic pain.

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.

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.

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.