Objective To investigate the effects of isoflurane on the cell cycle and apoptosis in PC12 cells.Methods The neuronal PC12 cells were cultured for 7 d with nerve growth factor in vitro.The cells were cultured in 25 cm2 culture flask and randomly divided into 2 groups (n=6 each): control group (group C) and isoflurane group (group Ⅰ).The neuronal PC12 cells were exposed to 1.2％ isoflurane for 12 h in group Ⅰ.The cell morphology was examined by light microscopy.The apoptotic rate,cell cycle progression,mitochondrial membrane potential (MMP) and intracellular Ca2 + concentrations ([Ca2 +]i) were assessed by flow cytometry.Results Compared with group C,the cell morphology was changed,the proportion of the cells in Go/G1 phase and MMP were significantly decreased,and the proportion of the cells in G2/M phase,apoptotic rate and [Ca2+]i were significantly increased in group Ⅰ (P ＜ 0.05).Conclusion Exposure of PC12 cells to 1.2％ isoflurane for 12 h can activate the cell cycle abnormally and result in cell apoptosis.

Objective To evaluate the efficacy and the effects of sevoflurane induced hypotension on balance of cerebral oxygen supply and consumption and on cerebral energy metabolism. Methods Forty patients undergoing elective procedures were randomly divided into three groups. Group Ⅰ :sevoflurane induced hypotension ; group Ⅱ: sevoflurane maintained normal arterial tension ; group Ⅲ: nitroprusside induced hypotension . In groupⅠ and Ⅲ MAP was decreased to 50%-60% of baseline ,was kept for 40 min either by sevoflurane inhalation or by nitroprusside infusion. MAP ,HR,ECG were monitored continuously, and radial arterial blood samples and jugular blood samples were taken synchronously for measuring blood gas and blood lactate ,SOD and MAD levels. Results In group Ⅰ during induced hypotension Da jvO 2 showed reduction ,but in group Ⅲ Da jvO 2 increased significantly. There were no significant changes in blood lactate level and in the cerebral arteriovenous differences of MAD and SOD in three groups.Conclusions The sevoflurane induced hypotension has no adverse effect on cerebral oxygen balance and the cerebral perfusion . The cerebral energy metabolism is well maintained . Sevoflurane can safely applied to controlled hypotension during cerebral neurosystem.

Objective To investigate the effects of propofol on gastric mucosal cellular apoptosis after resuscitation of hemorrhagic shock in rabbits.Methods One hundred healthy adult male New Zealand rabbits weighing 2.5-3.0 kg were randomly divided into 5 groups ( n =20 each):group sham operation (group S) ; group hemorrhagic shock ( group M ) and Ⅲ,Ⅳ,Ⅴ 3 propofol groups ( groups P1,2,3 ).Hemorrhagic shock was induced by withdrawing blood from femoral artery.MAP was reduced to 35-40 mm Hg and maintained at this level for 60 min in groups M,P1,P2 and P3.Blood was then transfused back via femoral vein to restore blood volume.In groups P1,2,3 propofol 5 mg/kg was injectel iv at 10 min before ischemia (group P1 ),10 min before (group P2 ) and 20 min of resuscitation (group P3 ) respectively followed by continuous infusion at 20 mg·kg-1 ·h-1 until 90 min of resuscitation.The gastric mucous membrane specimens were obtained at 90 min of resuscitation for macroscopic examination and detection of apoptosis (by TUNEL) and Bcl-2 and Bax protein expression (by immuno-histochemistry).Results Hemorrhagic shock seriously damaged gastric mucous membrane,significantly increased apoptotic index (the number of apoptotic cells/the total number of cells) and Bax protein expression and decreased Bcl-2 protein expression and Bcl-2/Bax ratio in group M as compared with group S.Propofol significantly attenuated hemorragic shock-induced above changes in groups P1 and P2.Concluion Pre- and post-conditioning with propofol can attenuate apoptosis in gastric mucous membrane cells induced by hemorrhagic shock by up-regulating Bcl-2 protein expression and down-regulating Bax protein expression.

OBJECTIVE: To establish a method for analysis of proteome of human peripheral leukocytes and to explore the difference between the healthy volunteers and patients with acute lymphocyte leukemia(ALL) in leukocyte proteomics and to study the ALL-related abnormal proteins.METHODS: Peripheral blood samples taken from 10 healthy volunteers and 10 ALL patients were separated for the extraction of leukocyte proteins.The leukocyte proteins were separated by two-dimensional electrophoresis(2-DE).2-DE patterns were analyzed by PDQuest 2D software and the differentially expressed proteins were identified by matrix assisted laser desorption ionization-time of flight-mass spectrometer(MALDITOF-MS) and bioinformatics.RESULTS: The 2-DE patterns of peripheral blood leucocytes from patients with ALL versus those from healthy volunteers were analyzed by PDQuest 2D software and the differentially expressed proteins identified by matrix assisted laser desorption ionization-time of flight-mass spectrometer(MALDI-TOF-MS) were confirmed to beALL-related proteins: AnnexinA5 and cytoskeleton 14 etc.CONCLUSION: Using MALDI-TOF-MS and bioinformatics,the AnnexinA5 and cytoskeleton 14 proteins had been proved to be of close relationship with ALL.The results provided a satisfactory basis for the searching for treatment targets and the molecular targets for the early diagnosis of ALL.

Objective To explore the effects of dexmedetomidine on analgesia of general anesthesia in elderly patients with lumbar vertebra surgery.Methods From August 2013 to for August 2014, 80 cases elderly patients with fracture of lumbar vertebra and prepared to have an surgery of pedicle screw internal fixation in Department of Anesthesiology in Beijing Jishuitan Hospital were admitted , and divided into control group and observation group.The control group were continuously-pumped with sufentanil and the observation group with dexmedetomidine for anesthesia.The anesthetic effect were compared between two groups.Results The rate of sedative satisfaction in observation group post-anesthesia was higher than that in control group (100.0%vs.75.0%;χ2 =11.429,P<0.05).The Ramesay score, awaking and extubation time perioperational period in observation group were lower than those in control group(P<0.05).The analgesic effect in observation group was higher than that in control group(100.0%vs. 90.0%;χ2 =4.211,P<0.05).Conclusion Dexmedetomidine has the obvious sedative and analgesic effect on elderly patients with lumbar vertebra surgery.

Objective To investigate the role of GABAB1 receptors in spinal dorsal horn neurons in the development of diabetic neuropathic pain (DNP). Methods Sixty pathogen free male SD rats aged 4 weeks weighing 150-170 g were randomly divided into 2 groups ( n = 30 each): control group and DNP group. Diabetes mellitus was induced by single intraperitoneal injection of streptozotocin 50 mg/kg. Blood glucose levels and paw withdrawal threshold to mechanical stimuli were measured at 3, 5 and 7 weeks (T1, T2, T3 ) after IP STZ/NS ( n = 10 each). The animals were sacrificed after PWL measurement. The lumbar segment of the spinal cord was removed for determination of the expression of GABAB1 receptors by immuno-histochemistry, RT-PCR and Western blot analysis. Results The blood glucose levels were significantly higher while the PWT was significantly lower at T1,T2 and T3 in group DNP than in control group. The expression of GABAB1 receptor mRNA and protein in spinal dorsal horn was significantly lower at T2 and T3 in DNP group than in control group. Conclusion The expression of GABAb1 receptors is down-regulated in spinal dorsal horn neurons in rats with DNP.

Objective To evaluate the role of calpain in sevoflurane anesthesia-induced apoptosis in hippocampal neurons of aged rats.Methods Fifty-four healthy female Sprague-Dawley rats,aged 18 months,weighing 450-550 g,were randomly divided into 3 groups (n=12 each) using a random number table:control group (group C),sevoflurane group (Sev group) and calpain inhibitor M DL28170 group (group M).In group C,the rats inhaled 50％ O2-50％N2 for 3 h.In Sev group,the rats inhaled 3％ sevoflurane for 3 h.In group M,MDL28170 10 mg/kg was injected via the tail vein,30 min later 3％ sevoflurane was inhaled for 3 h,and MDL28170 was simultaneously infused at 3.33 mg · kg 1 · h-1 via the tail vein.Nine rats in each group were selected,and cognitive function was assessed by using Morris water maze test at 30 min before anesthesia and 1-5 days after anesthesia.The escape latency and frequency of crossing the original platform were recorded.After the end of Morris water maze test performed at 30 min before anesthesia and 1-5 days after anesthesia,3 rats in each group were sacrificed,and hippocampal tissues were obtained for detection of cell apoptosis (by flow cytometry) and intracellular [Ca2+] i.Apoptotic rate was calculated.Results Compared with group C,the escape latency was significantly prolonged,and the frequency of crossing the original platform was decreased,and the apoptotic rate and intracellular [Ca2+]i were increased at 1 day after anesthesia in Sev and M groups.Compared with group Sev,the escape latency was significantly shortened,the frequency of crossing the original platform was increased,and the apoptotic rate was decreased at 1 day after anesthesia,and no significant change was found in intracellular [Ca2+]i in group M.Conclusion Calpain activation is involved in sevoflurane anesthesia-induced apoptosis in hippocampal neurons of aged rats.

Objective To evaluate the role of M3 muscarinic acetylcholine receptor (M3 mAChR) in the release of glycinergic neurotransmitter by using oxotremorine-M (Oxo-M: a nonselective mAChR agonist) and 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP: a highly selective M3mAChR antagonist). Methods Twenty male 3-4 weeks old SD rats weighing 160-180 g after successful intrathecal catheterization were randomized into 2 groups (n = 10 each): normal saline group (group NS) and pertussis toxin (group PTX).Pertussis toxin 1.5 μg/10 μl was injected IT in group PTX, while in group NS normal saline 10 μl was injected IT instead. The animals were killed at day 7 after injection. The spinal cords were removed and sliced and placed in artificial CSF. Glycinergic spontaneous inhibitory postsynaptic currents (sIPSCs) were measured in spinal lamina Ⅱneurons using whole-cell voltage-clamp technique. Five minutes after sealing, Oxo-M (final concentration 3 μ mol/L) was added. Oxo-M was then completely washed out 3 min later and 4-DAMP (final concentration 25 nmol/L) was added after 5 min of stabilization. In the presence of 4-DAMP, Oxo-M (final concentration 3 μmol/L) was added again 3 min later. sIPSCs were recorded before addition of Oxo-M (T1), 3 min after addition of Oxo-M (T2), 3 min after addition of 4-DAMP (T3), 3 min after the second addition of Oxo-M (T4). Results Compared with the baseline value at T1 , Oxo-M significantly increased the frequency of glycinergic sIPSCs at T2without changing the amplitude at T2-4 in both groups. The frequency of sIPSCs was significantly lower at T4 than at T2 in both groups (P ＜ 0.05 or 0.01). There was no significant difference in both frequency and amplitude of glycinergic sIPSCs between the two groups. Conclusion M3 mAChR plays a predominant role in the release of glycinergic transmitter in the spinal lamina Ⅱ neurons in rats.