Objective To investigate the effects of pretreatment with Shen-fu injection on mitochondrial permeability transition and mitochondrial transmembrane potential (△ψm) following myocardial ischemiareperfusion (IR) injury in rats. Methods Thirty SD rats of both sexes weighing 250-300 g were randomly divided into3 groups with 10 animals in each group:Ⅰ sham operation group (group S); Ⅱ IR group and Ⅲ Shen-fu injection group (group SFI). The animals were anesthetized with intraperitoneal 20% urethane 5 ml/kg. The chest was opened and the heart exposed. Myocardial IR was induced by temporary ligation of the anterior descending branch of left coronary artery maintained for 30 min, followed by 120 min reperfusion. Myocardial ischemia was confirmed by decoloration of apex and elevation of S-T segment (＞ 0.1 mV) or erection of T wave. In group SFI,SFI 10 ml/kg was infused at 15 min before ischemia, while in group S and IR equal volume of normal saline was infused instead of SFI. At 120 min of reperfusion, blood samples were collected from right internal carotid artery for determination of serum concentration of cTnI. The animals were then sacrificed and the hearts were immediately removed for measurement of myocardial mitochonerial permeability transition pore (MPTP) activity (by spectrophotometry at 540 nm) and myocardial △ψm (by fluorospectrophotometer using rhodamine 123 as fluorescent probe). Results Compared with group S, serum cTnI concentration and MPTP activity were significantly increased and △ψm was decreased in group IR. SFI pretreatment significantly attenuated the IRinduced increase in serum cTnI concentration and the MPTP activity and decrease in △ψm. Conclusion SFI pretreatment can protect myocardium from IR injury by attenuating the IR induced increase in MPTP opening and decrease in △ψm.

Objective To study the effect of dexmedetomidine(DEX)intrathecal administration on formation of chronic neuropathic pain induced by chronic constriction injury(CCI)in rats. Methods One hundred and twenty healthy adult male Sprague-Dawley(SD)rats were divided randomly into four groups(each n=30):sham group, model group,30μg/kg and 60μg/kg DEX intrathecal injection group(D30 group,D60 group). The CCI model was installed by left sciatic nerve ligaturing,sham surgery was done by exposing the sciatic nerve without ligation,and 30 μg/kg,60 μg/kg DEX(each,10 μl)and 10 μl normal saline were given intrathecally in D30,D60 and model groups respectively,all kinds of injection being once a day for 7 days. Hind paw mechanical contraction reflex threshold(MWT),heat-shrinkable reflex latency(TWL)and the score of motor dysfunction of hind extremity in rats were recorded on 1 day before ligation and 1,3,7 days after operation,and before ligation and 1,7,14 and 21 days after operation,the contents of tumor necrosis factor-α(TNF-α),interleukin(IL-1βand IL-6)in intumescentia lumbalis were measured by enzyme linked immunosorbent assay (ELISA). Results Compared with before operation,there were no significant differences in values of MWT,TWL,the motor function evaluation,TNF-α, IL-1β,and IL-6 at each time points(all P>0.05);however,with the prolongation of time,MWT and TWL were decreased,and the motor function evaluation,TNF-α,IL-1β,and IL-6 were increased in all the other groups. Compared with those of the sham group,MWT and TWL were declined,and the motor function evaluation,TNF-α, IL-1β,and IL-6 were elevated in model group. Compared with those of model group,30μg/kg and 60μg/kg DEX could significantly raise MWT(g)and TWL(s),obviously improve motor function and remarkably decrease the contents of TNF-α(pg/mg),IL-1β(pg/mg)and IL-6(pg/mg)in the spinal cord of CCI rats from 1 day after operation. And the changes in 60 μg/kg DEX group were more significant than those in 30 μg/kg DEX intrathecal injection group〔postoperative 1 day MWT:39.3±1.3 vs. 20.3±2.2,TWL:10.9±0.4 vs. 8.2±1.1,motor function score:2.00±0.00 vs. 2.00±0.75,TNF-α:33±7 vs. 125±18,IL-1β:108±12 vs. 203±34,IL-6:156±39 vs. 405±75,all P<0.05〕. Conclusions The DEX intrathecal administration has certain degree of dose-dependent therapeutic effect on hyperalgesia in CCI rat models. The mechanism is related to the amelioration of inflammatory reaction at the lumbar segment of spinal cord.

Objective To investigate the effects of post-propofol anesthesia on cognitive function and hippocampus proteome expressions in aged rats.Methods Thirty healthy male Wistar rats aged 20 months were randomly divided into control group(n =15) and propofol group(n =15).The control group was injected with normal saline of 6 ml/kg intraperitoneally and propofol was injected intraperitoneally with propofol 60 mg/kg.The rats in both groups underwent Step-down Test to assess cognitive function at the first day and at the seventh day after the termination of drug administration.Five rats were decapitated randomly each time after the two step-down tests and their hippocampi were removed for two-dimensional gel electrophoresis and mass spectrometric analysis.Results In the step-down test,aged rats in the propofol group showed significantly learning impairment and decreased memory abilities at the 1st day after propofol anesthesia as compared with those in the control group.In learning phase of the 1st day,the latency of the propofol group is (29.5 ± 7.6)s as compared with(19.7 ± 7.0)s of the control group,while the error time is 3.6±1.2 vs.1.6 ±0.8 in the propofol group vs the control group,and the total time of electric shock is(65.2 t 10.6)s vs.(42.7 ± 10.3)s in the propofol group vs the control group(all P＜0.01).The latency of the memory phase in the propofol group is also decreased as compared with that in the control group(31.4±14.3)s vs.(111.2± 23.7) s,(P＜0.01).On the 7th day after anesthesia,there was no significant difference between the two groups.There were 17 differentially expressed proteins on the 1st day after propofol anesthesia,6 of them were up-regulated and 11 proteins were down-regulated (P ＜ 0.05).On the 7th day,there were 10 differentially expressed proteins,and the expression of 5 proteins was down-regulated (P ＜ 0.01).Conclusions Aging rats receiving propofol anesthesia show cognitive function decline,but do not show a long-term decline.The mechanism may be related to the different expressions of hippocampal proteins.

Objective To evaluate the effects of inhalation sevoflurane in the early ischemia and reperfusion on pulmonary inflammatory response in patients undergoing cardiac surgery with extracorporeal circulation (ECC). Methods Forty patients with rheumatic heart disease scheduled for elective valve replacement were randomly assigned into 2 groups (20 patients in each group): control group and sevoflurane group. In sevoflurane group, 2% sevoflurane was inhaled for 15 min before and after the ascending aorta was blocked, and also before and after the ascending aorta was opened. Paitents in control group didn′t inhale sevoflurane. Time was defined as the followings: after anesthesia and before skin incision (T0), immediately before ECC (T1), immediately after ECC (T2), 2 h after ECC (T3), 6 h after ECC (T4) and 24 h after ECC (T5). At T0, T2, T3, T5, the radial artery blood was obtained to detect the levels of plasma tumor necrosis factor-α(TNF-α), interleukin-8(IL-8) and soluble intercellular adhesion molecule-1(sICAM-1). At T1, T2, the pulmonary artery and pulmonary vein blood was obtained to detect the neutrophil count and calculate the differences between the vein and artery. At T0, T2, T3, T4, T5, the arterial blood gas was detected and differences of alveoli-arterial oxygen pres [P(A- a)O2], oxygenation index (OI), static compliance (Cst) were calculated. Results The levels of plasma TNF-α, IL-8 and sICAM-1 were higher at T2, T3, T5 than those at T0 in two groups (P<0.05). The levels of plasma TNF-α, IL-8 and sICAM-1 were decreased in sevoflurane group at T2 and T3, compared with those in control group (P<0.05). The neutrophil counts of pulmonary artery, pulmonary vein and the differences between the vein and artery were higher at T2 than those at T0 in two groups (P<0.05). The neutrophil counts of pulmonary artery, pulmonary vein and the differences between the vein and artery were decreased in sevoflurane group at T2 compared with those of control group (P<0.05). The level of P(A- a)O2 was higher at T2, T3, T4 and T5 than that at T0 in two groups (P<0.05). The level of OI was decreased at T2, T3, T4 and T5 compared with that at T0 in two groups (P<0.05). The level of Cst was decreased at T2, T3 and T4 compared with that at T0 in two groups (P<0.05). The level of P(A-a)O2 was decreased in sevoflurane group at T2, T3 and T4 compared with that in control group (P<0.05). The levels of OI and Cst were higher in sevoflurane group at T2, T3 and T4 compared with those in control group (P<0.05). Conclusions Severe pulmonary inflammation often occurs during cardiac surgery with ECC, and it can be relieved by inhalation of sevoflurane in the early ischemia and reperfusion.

Objective To study the effect of propofol and midazolam on memory during target control infusion for sedation.Methods Forty healthy male volunteers were randomly devided into 4 groups,all subjects received target controlled infusion with propofol or midazolam for sedation.Group 1 and group 3 were sedated to OAA/S 1 or OAA/S 3 respectively by propofol;group 2 and group 4 was sedated to OAA/S 1 or OAA/S 3 by midazolam.All subjects learned a list of words before sedation and at the different sedation depths predicted.After recovery the explicit and implicit memory were tested using word stem completion method and process dissociation procedure.Results Compared with 0,there was no significant difference in explicit memory among all the 4 groups.While there was significant difference in implicit memory for group 3 and 4,and there was no significant difference in implicit memory for group1 and group2.Conclusion Sedative-hypnotic drugs not only deepen sedation,but also impair memory as serum concentration increases during target infusion.Drug-induced amnesia is independent of sedation.Propofol and midazolam have the same efficacy on memory at the same sedation scale.

Objective To investigate the role of Janus kinese 2-signal transducer and activator of transcription 3 (JAK2-STAT3) pathway in reduction of myocardial ischemia-reperfusion (I/R) injury by sufentanil postconditioning in dogs.Methods Twenty-four healthy dogs of either sex,weighing 10-15 kg,were randomly divided into 4 groups (n =6 each):sham operation group (group S); I/R group; sufentanil postconditioning group (group PO) and sufentanil postconditioning + specific JAK2 inhibitor AG490 group (group AG).In groups I/R,PO and AG,myocardial I/R was produced by occlusion of left anterior descending coronary artery for 30 min followed by 120 min reperfusion.In groups PO and AG,sufentanil 0.6 μg/kg was infused intravenously over 5 min before reperfusion and in addition in group AG,AG490 1 mg/kg was injected intravenously before sufentanil infusion.Myocardial specimens were taken at the end of 120 min reperfusion for microscopic examination and determination of the expression of caspase-3 and p-STAT3 by immuno-histochemistry and myocardial cell apoptosis index (AI) by TUNEL.Results AI and the expression of caspase-3 and p-STAT3 were significantly higher in groups I/R,PO and AG than in group S ( P ＜ 0.05).Compared with group I/R,AI and the expression of caspase-3 were significantly decreased in groups PO and AG,the expression of p-STAT3 was significantly increased in group PO,and the expression of p-STAT3 was significantly decreased in group AG ( P ＜ 0.05).AI and the expression of caspase-3 were significantly higher and the expression of p-STAT3 was significantly lower in group AG than in group PO (P ＜ 0.05).The pathologic changes were significantly attenuated in group PO compared with groups I/R and AG.Conclusion JAK2-STAT3 pathway is involved in reduction of myocardial I/R injury by sufentanil postconditioning in dogs.

Objective To investigate the mechanism underlying sufentanil postconditioning-induced reduction of myocardial ischemia-reperfusion (I/R) injury in rats through evaluating the relationship between phosphatidylinositol 3-kinase/serine-threonine kinase (PI3K/Akt) signaling pathway and mitochondrial permeability transition pore (mPTP).Methods Forty-eight pathogen-free healthy male SpragueDawley rats,aged 3 months,weighing 250-300 g,were divided into 4 groups (n=12 each) using a random number table:sham operation group (group S),group I/R,sufentanil postconditioning group (group SP) and snfentanil postconditioning plus PI3K inhibitor wortmannin group (group SP +W).The rats were anesthetized with 20％ urethane 5 ml/kg.Myocardial I/R was induced by ligation of the anterior descending branch of left coronary artery for 30 min,followed by 120 min reperfusion.Sufentanil 1.0 μg/kg was injected via the sublingual vein at 5 min before reperfusion in SP and SP+W groups.Wortmannin 15 pμg/kg was injected via the sublingual vein at 5 min before reperfusion,and then sufentanil 1.0 μg/kg was given in group SP+W.Blood samples were taken from the abdominal aorta at the end of reperfusion for detection of serum cardiac troponin I (cTnI) and creatine kinase-MB (CK-MB) concentrations.The rats were then sacrificed and hearts were removed for determination of cell apoptosis (by TUNEL),nicotinamide adenine dinueleotide (NAD+) content (by speetrophotometry),and expression of phosphorylated Akt (p-Akt) in myocardial tissues (by Western blot).Apoptosis index (AI) was calculated.The myocardial mitochondria and cytoplasm were isolated for detection of the expression of cytochrome c (Cyt c) and apoptosis-inducing factor (AIF) using Western blot.Results Compared with group S,the serum cTnI and CK-MB concentrations and AI were significantly increased,the content of NAD+ was decreased,the expression of p-Akt was up-regulated,the expression of Cyt e and AIF in mitochondria was down-regulated,and the expression of Cyt c and AIF in cytoplasm was up-regulated in I/R,SP and SP+W groups (P＜0.05).Compared with group I/R,the serum cTnI and CK-MB concentrations and AI were significantly decreased,the content of NAD+ was increased,the expression of p-Akt was up-regulated,the expression of Cyt c and AIF in mitochondria was up-regulated,and the expression of Cyt c and AIF in cytoplasm was down-regulated in group SP (P＜0.05).Compared with group SP,the serum cTnI and CK-MB concentrations and AI were significantly increased,the content of NAD+ was decreased,the expression of p-Akt was down-regulated,the expression of Cyt c and AIF in mitochondria was down-regulated,and the expression of Cyt c and AIF in cytoplasm was up-regulated in group SP+W (P＜0.05).Conclusion Sufentanil postconditioning can activate PI3K/Akt signaling pathway,inhibit mPTP opening,mitigate mitochondrial injury and inhibit apoptosis in cardiomyocytes,thus attenuating myocardial I/R injury in rats.

Objective To evaluate the effect of sufentanil postconditioning on the level of cathepsin B in the myocardium during ischemia-reperfusion (I/R) in the rats.Methods Eighteen healthy adult male Sprague-Dawley rats,weighing 250-300 g,were divided into 3 groups (n=6 each) using a random nunber table:shamn operation group (group S),group I/R and sufentanil postconditioning group (group SP).The rats were anesthetized with intraperitoneal 20％ urethane 5 ml/kg.Myocardial I/R was induced by occlusion of the left anterior descending branch of the coronary artery for 30 min followed by 120 min reperfusion.At 5 min before reperfusion,sufentanil 1.0 μg/kg was intravenously injected in group SP,and the equal volume of normal saline was given instead in S and I/R groups.At the end of reperfusion,blood samples from the abdominal aorta were collected for determination of serum cardiac troponin I (cTnI) and creatine kinase-MB (CK-MB) concentrations,and myocardial specimnens were obtained for examination of the ultrastructure of cardiomyocytes (using transmission electron microscopy) and for determination of Beclin1,microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ) and cathepsin B expression (by Western blot) and cathepsin B activity (by fluorometric assay).Results Compared with group S,the serum cTnI and CK-MB concentrations were significantly increased,the expression of Beclin-1,LC3 Ⅱ and cathepsin B was up-regulated,and the activity of cathepsin B was enhanced in I/R and SP groups (P＜0.05).Compared with group I/R,the serum cTnI and CK-MB concentrations were significantly decreased,the expression of Beclin-1 and LC3 Ⅱ was down-regulated,the expression of cathepsin B was up-regulated,and the activity of cathepsin B was enhanced (P＜0.05),the pathological changes of myocardial tissues were signifieantly attenuated,and autophagosomes were reduced in group SP.Conclusion The mechanism by which sufentanil postconditioning inhibits cardiomyocyte autophagy during myocardial I/R is probably related to increased expression and activity of cathepsin B in rats.

[ ABSTRACT] AIM: To investigate the protective effect of autophagy on oxidized low density lipoprotein ( ox-LDL)-induced macrophage apoptosis and the underlying molecular mechanisms .METHODS:The RAW264.7 macropha-ges were pretreated with 3 mmol/L 3-methyladenine (3-MA), 1 μmol/L rapamycin (Rap) or 4 mmol/L 4-phenylbutyric acid ( PBA) respectively for 1 h and then treated with ox-LDL (100 mg/L) for 12 h.The cell viability and apoptosis were determined by MTT assay and flow cytometry with Annexin V-FITC/PI staining, respectively.The activities of lactate de-hydrogenase ( LDH) in the medium and caspase-3 in the cells were determined by detection kits .The protein levels of bec-lin-1 (a molecular marker of autophagy ), glucose-regulated protein 78 (GRP78, an endoplasmic reticulum stress marker) and C/EBP homologous protein ( CHOP, a key-signaling component of endoplasmic reticulum stress-induced apoptosis ) were examined by Western blot .Microtubule-associated protein 1 light chain 3 (LC3, another molecular marker of autoph-agy) was observed under laser scanning confocal microscope .RESULTS: Treatment of the RAW264.7 macrophages with ox-LDL at 100 mg/L for 12 h resulted in significant decrease in cell viability , and dramatic elevation in LDH leakage , cell apoptosis and caspase-3 activity, which were promoted by 3-MA (an autophagy inhibitor) and inhibited by Rap (an autoph-agy inducer ) .ox-LDL induced autophagy in the macrophages as assessed by beclin-1 upregulation and frequent granulation of LC3, which were inhibited by 3-MA and promoted by Rap.Interestingly, 3-MA enhanced, while Rap blocked, the CHOP upregulation induced by ox-LDL.Moreover , PBA ( endoplasmic reticulum stress inhibitor ) significantly inhibited ox-LDL-induced GRP78 upregulation and autophagy as determined by the attenuation of beclin-1 upregulation and frequent granula-tion of LC3.CONCLUSION: Endoplasmic reticulum stress mediates ox-LDL-induced autophagy in macrophages , and moderates activation of autophagy may protect macrophages from ox-LDL-induced apoptosis by inhibiting CHOP expression .

Objective:To analyze the clinical characteristics and influencing factors on postoperative emotional and cognitive function for elderly versus non-elderly male patients with laryngeal carcinoma.Methods:The patients with laryngeal cancer hospitalized in the Department of Otorhinolaryngology-Head and Neck Surgery were selected for a questionnaire survey in two Grade III-A Hospital in Shanxi Province from January 2018 to December 2019.There were 105 patients with laryngeal cancer, including 60 in the elderly and 45 in the non-elderly group.Negative emotion and cognitive function were investigated by using Self-rating Anxiety Scales(SAS), Self-rating Depression Scale(SDS), and Montreal Cognitive Assessment(MOCA)before surgery, 10 days after surgery, and 1 year after surgery, respectively.The clinical characteristics of the elderly versus non-elderly groups were analyzed and compared.Results:One year after surgery, there were 52 cases(86.7%)and 27 cases(45.0%)of depression and cancer-related cognitive impairment(CRCI)in the elderly group, which were higher than 30 cases(66.7%)and 4 cases(8.9%)in the non-elderly group, with statistically significant difference( χ2 = 6.013, χ2 =16.115 and P<0.05, P<0.01). The elderly group showed much more anxiety and CRCI 10 days after surgery(60.50±4.31 vs.55.84±3.81, 27.47±1.08 vs.28.31±1.08, P<0.01)and showed much more depression and CRCI(57.20±5.66 vs.62.60±5.37, 27.36±1.37 vs.26.08±1.42, P<0.01)than did the non-elderly group 1 year after surgery.The results of multiple linear regression analysis in the elderly group showed that the mode of operation and the pronunciation one year after surgery significantly affected their anxiety; the marriage and the educated level significantly affected their depression; the marriage, the educated level and operation mode significantly affected their cognitive function.Clinical stage of tumor significantly affected anxiety and depression, the educated level significantly affected cognitive function in the non-elderly group. Conclusions:Postoperative negative emotion and cognitive function in patients with laryngeal cancer continues to deteriorate after surgery, which were affected by many factors, especially in elderly patients.It is necessary to conduct active and effective interventions as soon as possible.