Role of TRPV4 in dexmedetomidine-induced improvement in cognitive function in mice with mechanical ventilator-caused brain injury / 中华麻醉学杂志

Objective:To evaluate the role of transient receptor potential vanillic acid 4 (TRPV4) in dexmedetomidine-induced improvement in cognitive function in mice with mechanical ventilator-caused brain injury.Methods:Ninety clean-grade healthy male C57BL6 mice, weighing 20-25 g, aged 8-12 weeks, were divided into 5 groups ( n=18 each) using a random number table method: control group (group C), mechanical ventilation group (group V), HC-067047 group (group H), dexmedetomidine group (group D), and dexmedetomidine+ GSK1016790A group (group DG). In group C, the animals breathed air spontaneously for 6 h without mechanical ventilation. In group V, the animals were mechanically ventilated for 6 h. In group H, TRPV4 blocker HC-067047 10 mmol was injected into the cerebral ventricle at 3 and 6 h of mechanical ventilation. In D and DG groups, dexmedetomidine 50 μg/kg was intraperitoneally injected at 30 min before mechanical ventilation. In group DG, TRPV4 agonist GSK1016790A 5 μmol was injected into the cerebral ventricle at 60 min before mechanical ventilation. Morris water maze test was performed on 6 mice in each group at 1 day before mechanical ventilation and 3 and 7 days after mechanical ventilation. Six mice in each group were randomly selected and sacrificed at 1 day after mechanical ventilation, and the brain tissue was taken for determination of the neuronal apoptosis in hippocampal CA1 area by TUNEL method, and the apoptosis index was calculated. Six mice in each group were randomly selected and sacrificed at 1 day after mechanical ventilation, and the hippocampal tissues were taken for determination of the expression of TRPV4, serine-threonine protein kinase (Akt), phosphorylated Akt (p-Akt), Bcl-2, Bax and caspase-3 by Western blot. Results:Compared with group C, the escape latency was significantly prolonged and the number of crossing the original platform was reduced at 3 and 7 days after mechanical ventilation, the expression of TRPV4 and caspase-3 was up-regulated, the ratio of Bcl-2/Bax was decreased, and the apoptosis index of neurons was increased in group V and group DG ( P<0.05). Compared with group V, the escape latency was significantly shortened and the number of crossing the original platform was increased at 3 and 7 days after mechanical ventilation, the expression of TRPV4 and caspase-3 was down-regulated, the expression of p-Akt was up-regulated, the ratio of Bcl-2/Bax was increased, and the apoptosis index of neurons was decreased in group D and group H ( P<0.05). Compared with group D, the escape latency was significantly prolonged at 3 and 7 days after mechanical ventilation, the number of crossing the original platform was reduced, the expression of TRPV4 and caspase-3 was up-regulated, the expression of p-Akt was down-regulated, the ratio of Bcl-2/Bax was decreased, and the apoptosis index of neurons was increased in group DG ( P<0.05). Conclusions:TRPV 4 is involved in dexmedetomidine-induced improvement in cognitive function, which is related to up-regulation of p-Akt expression and inhibition of apoptosis in hippocampal neurons in mice with mechanical ventilation-caused brain injury.

A case report of primary extubation by partial cricotracheal resection for severe subglottic stenosis / 临床耳鼻咽喉头颈外科杂志

This patient suffered from severe subglottic stenosis(grade Ⅳb). During partial cricotracheal resection, we cut through the cricothyroid membrane and the cricoid arch along the line from the lower edge of the thyroid cartilage to 5 mm of the inferior thyroid cartilage corner anteromedially. This can protect the cricothyroid joint, effectively protect the recurrent laryngeal nerve, and also support the airway. Strictly adhere to airway separation, avoid excessive separation of scars, and combine with reasonable postoperative management to achieve a safe extubation.

Role of TRPV1/NF-κB signaling pathway in dexmedetomidine-induced alleviation of VILI in rats / 中华麻醉学杂志

Objective:To evaluate the role of transient receptor potential vanilloid receptor 1 (TRPV1)/nuclear factor-κB (NF-κB) signaling pathway in dexmedetomidine-induced alleviation of ventilator-induced lung injury (VILI) in rats.Methods:One hundred clean-grade healthy male Sprague-Dawley rats, weighing 270-320 g, aged 4-5 months, were divided into 5 groups ( n=20 each) using a random number table method: control group (group C), VILI group (group V), AMG9810 group (group A), dexmedetomidine group (group D), and dexmedetomidine + RTX group (group DR). VILI model was prepared by mechanical ventilation with a tidal volume of 40 ml/kg for 4 h. In group A, TRPV1 inhibitor AMG9810 30 mg/kg was intraperitoneally injected at 1 h before mechanical ventilation.Dexmedetomidine 5.0 μg/kg was intravenously infused at 20 min before mechanical ventilation, and dexmedetomidine was intravenously infused at the rate of 5.0 μ g·kg -1·h -1 during ventilation in group D and group DR.In group DR, RTX 70 μ g/kg was intraperitoneally injected for 3 consecutive days before mechanical ventilation.At 4 h of mechanical ventilation, the concentrations of interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and IL-6 in bronchoalveolar lavage fluid (BALF) were detected, oxygenation index (OI) and wet/dry lung weight (W/D) ratio were measured, the histopathological changes of lung tissues were observed, and lung injury was assessed and scored.The expression of TRPV1 and NF-κB in lung tissues was detected by Western blot, and real-time polymerase chain reaction was used to detect the expression of TRPV1 and NF-κB mRNA. Results:Compared with group C, the concentrations of IL-1β, TNF-α and IL-6 in BALF were significantly increased, OI was decreased, the W/D ratio and lung injury scores were increased, and the expression of TRPV1 and NF-κB protein and mRNA was up-regulated in group V ( P<0.05). Compared with group V, the concentrations of IL-1β, TNF-α and IL-6 in BALF were significantly decreased, OI was increased, the W/D ratio and lung injury scores were decreased, and the expression of TRPV1 and NF-κB protein and mRNA was down-regulated in A, D and DR groups ( P<0.05). Compared with group D, the concentrations of IL-1β, TNF-α and IL-6 in BALF were significantly increased, OI was decreased, the W/D ratio and lung injury scores were increased, and the expression of TRPV1 and NF-κB protein and mRNA was up-regulated in group DR ( P<0.05). Conclusions:The mechanism by which dexmedetomidine alleviates VILI is partially related to inhibition of the activation of TRPV1/NF-κB signaling pathway and inhibition of the inflammatory responses in lung tissues of rats.

Effects of dexmedetomidine on alveolar epithelial barrier function in rats with VILI and the role of PKC / 中华麻醉学杂志

Objective:To evaluate the effects of dexmedetomidine on alveolar epithelial barrier function in rats with ventilator-induced lung injury (VILI), and the role of protein kinase C (PKC).Methods:One hundred clean-grade male Sprague-Dawley rats, weighing 270-320 g, aged 4-5 months, were divided into 5 groups ( n=20 each) using a random number table method: control group (group C), VILI group (group V), PKC inhibitor group (group B), dexmedetomidine group (group D), and dexmedetomidine plus PKC agonist group (DP group). The VILI model was developed by mechanical ventilation with a tidal volume of 40 ml/kg for 4 h in anesthetized animals.Group C breathed air autonomously for 4 h without mechanical ventilation.Group V was mechanically ventilated for 4 h. In group B, bisindolvlmaleimide I 0.12 mg/kg was injected intramuscularly 1 h before mechanical ventilation.In D and DP groups, dxmedetomidine 5.0 μg/kg was injected intravenously at 20 min before mechanical ventilation, and dexmedetomidine was intravenously infused at the rate of 5.0 μg·kg -1·h -1 during mechanical ventilation.In group DP, PKC agonist phorbol-12-myristic acid-13-acetate 15 μg/kg was intraperitoneally injected at 30 min before mechanical ventilation.At 4 h of mechanical ventilation, oxygenation index (OI), lung permeability index (LPI) and wet/dry lung weight (W/D) ratio were measured, the pathological changes of lung tissues were observed, and lung injury was assessed and scored.The expression of PKC, occludin and ZO-1 protein was detected by Western blot, and the expression of PKC mRNA, occludin mRNA and ZO-1 mRNA was determined by real-time polymerase chain reaction. Results:Compared with group C, OI was significantly decreased, LPI, W/D ratio and lung injury score were increased, the expression of PKC protein and mRNA was up-regulated, and the expression of occludin and ZO-1 protein and mRNA was down-regulated in V and DP groups ( P<0.05), and no significant change was found in the parameters mentioned above in B and D groups ( P>0.05). Compared with group V, OI was significantly increased, LPI, W/D ratio and lung injury score were decreased, the expression of PKC protein and mRNA was down-regulated, and the expression of occludin and ZO-1 protein and mRNA was up-regulated in B, D and DP groups ( P<0.05). Compared with group D, OI was significantly decreased, LPI, W/D ratio and lung injury score were increased, the expression of PKC protein and mRNA was up-regulated, and the expression of occludin and ZO-1 protein and mRNA was down-regulated in group DP ( P<0.05). Conclusions:Dexmedetomidine can reduce the damage to alveolar epithelial barrier function in rats with VILI, and the mechanism is related to inhibition of PKC activation and up-regulation of the expression of occludin and ZO-1.

Construction of clinical scoring system for predicting microvascular invasion in preoperative hepatocellular carcinoma / 中华普通外科杂志

Objective:To analyze the risk factors of hepatocellular carcinoma microvascular invasion (MVI) and to construct a preoperative prediction clinical scoring system.Methods:A retrospective analysis was made on 113 patients with hepatocellular carcinoma undergoing hepatectomy at Zhongshan Hospital from March 2018 to Jun 2019.Postoperative pathology confirmed 35 cases with microvascular invasion.Results:The multivariate logistic regression model showed that the maximum tumor diameter( OR: 1.028, 95% CI: 1.001-1.005), the smoothness of the capsule edge( OR: 0.208, 95% CI: 0.062-0.699), the positive circulating tumor cells (CTC)( OR: 3.728, 95% CI: 1.029-13.501) and abnormal prothrombin(PIVKA-Ⅱ)( OR: 1.001, 95% CI: 1.000-1.002) were risk factors for MVI. The area, sensitivity and specificity of the clinical score constructed by assigning 1 point to each risk factor were 0.906, 74.29% and 92.31%, respectively. Clinical scores of 0, 1, 2, 3, and 4 predict MVI positive rates of 0 (0/26), 9.09% (3/33), 28.57% (6/21), 77.78% (14/ 18), 85.71% (12/14). Conclusions:Tumor maximum diameter>62 mm, PIVKA-Ⅱ>115 mAU/ml, unsmooth tumor capsule and CTC in peripheral blood are independent high risk factors in patients with MVI.

Relationship between mechanism underlying methylprednisolone-induced alleviation of ventilator-induced lung injury and p38MAPK/NLRP3 pathway in lung tissues of rats / 中华麻醉学杂志

Objective:To evaluate the relationship between the mechanism underlying methylprednisolone-induced alleviation of ventilator-induced lung injury (VILI) and p38 mitogen-activated protein kinase (p38 MAPK)/nucleotide binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) pathway in lung tissues of rats.Methods:Sixty clean-grade male Sprague-Dawley rats, weighing 270-320 g, aged 4-5 months, were divided into 3 groups ( n=20 each) using a random number table method: control group (group C), mechanical ventilation group (group V), and methylprednisolone group (group M). Group C breathed air spontaneously for 4 h without mechanical ventilation.Group V was mechanically ventilated (RR 40 times/min, V T 40 ml/kg, I∶E 1∶1, PEEP 0, FiO 2 21%) for 4 h. Group M received intravenous methylprednisolone 10 mg/kg at 20 min before mechanical ventilation.At 4 h of mechanical ventilation, broncho-alveolar lavage fluid (BALF) was collected to measure the concentrations of interleukin-1beta (IL-1β), IL-18, and tumor necrosis factor-alpha (TNF-α) and wet/dry lung weight ratio (W/D ratio), and lung tissues were obtained for microscopic examination of the histopathological changes and for detection of the expression of p38MAPK, phosphorylated p38MAPK (p-p38MAPK), NLRP3, apoptosis-related speck-like protein containing a CARD (ASC), and cysteinyl aspartate-specific protease-1 (caspase-1) (using Western blot). Results:Compared with group C, the W/D ratio of lung tissues and concentrations IL-1β, IL-18 and TNF-α in BALF were significantly increased, and the expression of p-p38MAPK, NLRP3, ASC and caspase-1 was up-regulated in group V ( P<0.05), and no significant change was found in group M ( P>0.05). Compared with group V, the W/D ratio of lung tissues and concentrations of IL-1β, IL-18 and TNF-α in BALF were significantly decreased, and the expression of p-p38MAPK, NLRP3, ASC and caspase-1 was down-regulated in group M ( P<0.05). Conclusion:The mechanism by which methylprednisolone alleviates VILI may be related to inhibition of p38MAPK/NLRP3 pathway activity and reduction of inflammatory responses in lung tissues of rats.

Prevention experience of biliary tract complications after liver transplantation from organ donation after citizen's death / 器官移植

Objective To summarize the experience of prevention of biliary tract complications after liver transplantation from organ donation after citizen's death. Methods Clinical data of 88 cases undergoing liver transplantation from organ donation after citizen's death in the Affiliated Zhongshan Hospital of Sun Yat-sen University from October 2008 to December 2016 were retrospectively analyzed. Results Eighty-eight cases were eligible for the standards for organ donation after brain death plus cardiac death according to the Ⅲ national system for organ donation in China. According to the standard procedures, donor livers were successfully harvested and transplanted in 88 recipients. The biliary tract was reconstructed using the bile duct end-to-end anastomosis. The length of bile duct in the donors was shortened as possible. Slight tension should be maintained during anastomosis. Neither primary liver graft nonfunction nor rejection reaction occurred. One recipient suffered from bile leakage and recovered after drainage for 3 weeks. Two patients presented with biliary tract stenosis and mitigated after the placement of biliary tract stent. Conclusions The harvesting of donor liver should be in accordance with the standard procedures. The advantages of extracorporeal membrane oxygenation (ECMO) should be fully utilized to shorten warm and cold ischemia time as possible. Much attention should be diverted to the reconstruction of biliary tract, which contributes to decreasing the risk of biliary tract complications. Favorable clinical efficacy can be achieved in liver transplantation from organ donation after citizen's death.

Construction of a new alcoholic liver disease mouse model / 上海交通大学学报（医学版）

Objective·To establish a reliable alcoholic liver disease mouse model (ALDNM) that mimics the drinking pattern of alcoholic liver disease (ALD) patients.Methods·Using the self-designed feeding tubes and liquid diet,ALDNM model was developed through chronic feeding combined with acute gavage of ethanol based on Lieber-DeCarli model and Gao-Binge model.C57BL/6 mice were administered with control liquid diet for adaptation for first 5 d,and then divided into pair-fed group and ethanol-fed group (10 mice each group).Ethanol-fed mice were fed with the liquid diet in which ethanol accounts for 30％ of total energy,while the pair-fed mice were fed with the control diet for 10 d.At the 16th day,ethanol-fed mice and pair-fed mice were respectively gavaged a single dose of 31.5％ ethanol or isocaloric maltose dextrin,and euthanized 9 h later.Sera and livers were collected.The general physiological condition,hepatic tissue pathological changes and serum indexes between Lieber-DeCarli models and ALDNM models were compared.The liver lipids of ALDNM mice were determined by Oil red O (ORO) staining and hepatic triacylglyceride (TAG) test.Meanwhile,the mRNA levels of interleukin-6 (IL-6),tumor necrosis factor α (TNF-α),fatty acid synthase (Fas),long chain fatty acid elongase 6 (Elovl6) and stearyl-CoA desaturase (Scdl) were detected by real-time PCR in ALDNM models.Western blotting was used to detect the changes of phosphorylated signal transduction and transcriptional activator (p-STAT3) in the livers.Results·Lieber-DeCarli model mice were generally in poor condition,and there was no significant change in serum glutamic-pyruvic transaminase (GPT) and glutamic-oxaloacetic transaminase (GOT) compared to pair-fed group.However,in ALDNM models,H-E staining showed that the hepatocytes of ethanol-fed mice were extremely swollen with round volume,increased cytoplasm and filled with large amounts of fat vacuoles.ORO staining analyses showed obvious microsteatosis in the liver cells from all ethanol-fed mice.The hepatosomatic index,liver TAG content,serum GPT and GOT of ALDNM models were significantly higher than those in the pair-fed group,while the serum HDL significantly decreased compared to the pair-fed group.Moreover,the expression levels of both lipid synthesis pathways and inflammatory signaling pathways related genes in livers significantly increased in the ethanol-fed mice of ALDNM model.Conclusion·ALDNM model was successfully constructed.This model is cost-and time-efficient.Moreover,ALDNM model mimics the drinking pattern and pathogenesis of ALD patients with the advantages of stable food intake,good repeatability,and obvious liver damage.

The Rehabilitation Effects of the Cochlear Implantation on Prelingually Deaf Children with Alba Abnormality / 听力学及言语疾病杂志

Objective To study the rehabilitation effects of the cochlear implants on prelingually deaf children with alba abnormality.Methods A retrospective analysis of the effects of CIs was conducted in the prelingually deaf children of 11 cases of the children with abnormal alba(the research group) and 18 cases of the children who had normal alba(the control group) at the First Affiliated Hospital of Fujian medical university.All the operations were completed by the same doctor.There were no obvious complications during and after the operation.The cochlear implants were turned on after one month and the prelingually deaf children with extremely severe bilateral sensorineural deafness were trained for speech at the rehabilitation centre.The assessment criteria of the categories of auditory performance(CAP) and speech intelligibility rate(SIR) were used.After six months and twelve months of the operation, the family members were followed who have direct contacts with the children.The evaluation of data between the research and the control groups was administered.Results In the research group, the average level of CAP after six months'' post-operation was 2.41±0.47.But in the control group, the average level was 3.28±0.45.In the research group, the average level of SIR after six months'' post-operation was 1.27±0.44.There were 3 children in the research group at level 2, but in the control group, the average level was 1.89±0.31.Two children in the control group were level 1 while the others were level 2.In the research group, the average level of CAP after twelve months'' post-operation was 4.00±0.43 while only one child at level 3.There was one child at level 5 in the research group, the rest were level 4.There was a statistically significant difference in the average level of the CAP after six months'' post-operation(t=4.983, P0.05) and in the average level of the SIR after twelve months'' post-operation(t=0.434, P>0.05).Conclusion There is no significant difference of the rehabilitation effects of post-operation between the prelingually deaf children with alba abnormality and those with normal alba.

Knockdown of TRIM27 expression regulates cell proliferation, invasion and migration in human nasopharyngeal 5-8F carcinoma cells / 中国病理生理杂志

AIM:To investigate the expression characteristics of TRIM 27 in human nasopharyngeal carcinoma tissues, nasopharyngeal carcinoma 5-8F cells and NP69 cells, and to observe the effect of TRIM27 on the proliferation, in-vasion and migration of 5-8F cells.METHODS:The levels of TRIM27 in the nasopharyngeal carcinoma tissues and normal nasopharyngeal epithelial tissues were observed by the method of immunohistochemistry .The mRNA and protein levels of TRIM27 in the 5-8F cells and NP69 cells were determined by real-time PCR and Western blot .TRIM27 siRNA was trans-fected into the 5-8F cells with Lipofectamine 2000.The relative mRNA expression of TRIM27 was detected by real-time PCR.The relative protein expression of TRIM 27 was detected by Western blot .The cell proliferation was analyzed by CCK-8 assay and cell colony formation assay .The change of cell invasion was examined by Matrigel invasion assay .The change of cell migration were examined by wound healing assay .RESULTS:The results of immunohistochemistry showed that the protein expression of TRIM27 in the nasopharyngeal carcinoma tissues was obviously higher than that in the normal nasopha -ryngeal epithelial tissues .The results of real-time PCR and Western blot showed that the mRNA and protein levels of TRIM27 in the 5-8F cells were obviously higher than those in the NP69 cells.The abilities of proliferation, invasion and migration in the 5-8F cells were significantly suppressed after TRIM27 gene silencing ( P <0.05).CONCLUSION:TRIM27 acts as a oncogene in the 5-8F nasopharygeal carcinoma cells .The abilities of proliferation , invasion and migration are significantly suppressed after TRIM27 gene silencing in the 5-8F cells.

Effects of Puerarin on Bone Mineral Density around the Femoral Prosthesis of Elderly Women after Osteopo-rotic Fracture Artificial Hip Joint Replacement / 中国药房

OBJECTIVE:To investigate the effects of puerarin on bone mineral density around the femoral prosthesis of el-derly women after osteoporotic fracture artificial hip joint replacement. METHODS:99 elderly women after osteoporotic fracture artificial hip joint replacement were divided into control group(49 cases)and test group(50 cases)according to random number table. Control group received conventional treatment:calcium carbonate and vitamin D3+alendronate sodium+salmon calcitonin;test group was additionally given Puerarin injection 200-400 mg dissolved in Glucose injection 500 ml intravenously,qd,on the basis of control group. A treatment course lasted for 20 d,and both groups received 2 courses of treatment. The hip joint function score and bone mineral density around the femoral prosthesis of 2 groups were observed and compared after surgery,and the oc-currence of ADR was also observed. RESULTS:3 and 2 patients withdrew from control group and test group,respectively. 18 months after surgery,the patients with hip joint function score ranged 70-79 in test group was significantly less than in control group;the rate of excellent hip joint function score in test group was significantly higher than in control group,with statistical significance (P0.05);bone mineral density in R6-R7 range was signifi-cantly higher than control group(89.58% vs. 69.57%),with statistical significance(P<0.05). The prosthesis loosening was not found in both groups,and ADR was also not found as fever,erythra,nausea,vomiting,headache,dizziness,etc. CONCLU-SIONS:For the use of puerarin in elderly women after osteoporotic fracture artificial hip joint replacement,puerarin can increase the periprosthetic femur bone mineral density with good safety.

Effect of different concentrations of sevoflurane on neurotoxicity of primary cultured rat cortical neurons / 临床麻醉学杂志

Objective To explore the relationship of sevoflurane neurotoxicity with the expres-sion of Bid,Bim,Puma.Methods The cortical neuron from newborn SD rat (within 24 h)were see-ded in 6 or 12 well plate,and then randomly divided into 4 groups.Rat culture cortical neurons in vitro exposed in 1%,2%,4% and 0% sevoflurane for 6h were divided into A,B,C and D group. The effect of neuron viability,death and apoptosis were assessed using CCK-8,LDH and caspase-3 cleavage 1 7kDa expression assay.The expressions of Bid,Bim and Puma were assessed by western blot.Results Compared with group D, there were significant increases of neuron death and apoptosis,but a decrease of neuron viability,and upregulated expressions of Bid,Bim and Puma in group B (P <0.05);Compared with group B,Group C had increased death and apoptosis and de-creased viability of neurons,as well as upregulated expressions of Bid,Bim and Puma (P <0.05 ). Conclusion Along with the increase of the concentration,sevoflurane neurotoxicity was increased by upregulation of Bid,Bim,Puma expression.

Effects of Learning Environment on Learning and Memory and Microvessel Density in Hippocampus of Rats after Cerebral Infarction / 中国康复理论与实践

@# Objective To study the effects of learning environment on learning memory and microvessel density in hippocampus of rats after cerebral infarction. Methods 80 Sprague-Dawley rats were coagulated right middle cerebral arteries electrically, and randomly divided into learning envionment (LE) and standard environment (SE) groups. They were tested with Morris Water Maze 7 d and 28 d after operation. The microvessel density in hippocampus was measured with CD34 immunohistochemstry 1, 3, 7, 14 and 28 days after operation. Results The achievement of Morris Water Maze was significantly better in the LE group than in the SE group (P<0.001), while the microvessel density in the hippocampus was more since 7 days after operation (P<0.05). Conclusion The learning environment can promote the angiogenesis and improve the learning and memory function recovery.

Role of HIF-1α in reduction of apoptosis in cortical neurons of rats by sevoflurane preconditioning: the relationship with Bid, Bim and Puma / 中华麻醉学杂志

Objective To evaluate the role of hypoxia inducible factor-1α (HIF-1α) in reduction of apoptosis in cortical neurons of rats by sevoflurane preconditioning.Methods Primary cortical neurons obtained from neonatal Sprague-Dawley rats were seeded in 6-well plates (2 ml/well),and randomly divided into 4 groups (n =15 each) using a random number table:control group (C group),anoxiareoxygenation (A/R) group,sevoflurane preconditioning group (SP group) and HIF-1α inhibitor 2-methoxyestradiol group (H group).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h.In group SP,the neurons were exposed to 2.0％ sevoflurane for 2 h followed by 5 min washout for 3 times,and then sevoflurane preconditioning was performed immediately.In group H,sevoflurane preconditioning was performed at 72 h of incubation with 5 μmol/L 2-methoxyestradiol.The apoptosis in neurons was assessed using Annexin V-FITC/PI assay,and apoptosis rate was calculated.The expression of Bid,Bim,Puma and activated caspase-3 in neurons was detected by Western blot.Results Compared with group C,apoptosis rate was significantly increased,and the expression of Bid,Bim,Puma and activated caspase-3 was up-regulated in group A/R.Compared with group A/R,apoptosis rate was significantly decreased,and the expression of Bid,Bim,Puma and activated caspase-3 was down-regulated in group SP.Compared with group SP,apoptosis rate was significantly increased,and the expression of Bid,Bim,Puma and activated caspase-3 was up-regulated in group H.Conclusion HIF-1α mediates reduction of apoptosis in rat neurons by sevoflurane preconditioning,and down-regulated expression of Bid,Bim,and Puma is involved in the mechanism.

Role of HIF-1αin reduction of apoptosis in cortical neurons of rats by sevoflurane preconditioning:the relationship with Slit2∕Robo signaling pathway / 中华麻醉学杂志

Objective To evaluate the role of hypoxia inducible factor?1α ( HIF?1α) in reduction of apoptosis in cortical neurons of rats by sevoflurane preconditioning and the relationship with Slit2∕Robo signaling pathway. Methods Primary cortical neurons obtained from neonatal Sprague?Dawley rats were seeded in 6?well (2 ml∕well) or 96?well plates (100 μl∕well) at a density of 1×106∕ml, and randomly divided into 4 groups ( n=24 each ) using a random number table: control group ( C group ) , anoxia?reoxygenation ( A∕R ) group, sevoflurane preconditioning group ( SP group ) and HIF?1α inhibitor 2?methoxyestradiol group ( H group ) . The neurons were subjected to O2?glucose deprivation for 90 min followed by restoration of O2?glucose supply for 24 h. In group SP, the neurons were exposed to 2%sevoflurane for 2 h followed by 5 min washout with phosphate buffered saline for 3 times, and then sevoflurane preconditioning was performed immediately. In group H, sevoflurane preconditioning was performed with 5μmol∕L 2?methoxyestradiol at 72 h of incubation. The apoptosis in neurons was assessed using AnnexinⅤ?FITC∕PI assay, and apoptosis rate ( AR) was calculated. The amount of lactic dehydrogenase ( LDH) released was measured using colorimetric method. The expression of Slit2, Robo1 and Robo4 mRNA and protein was detected by fluorescent quantitative real?time polymerase chain reaction or Western blot. Results Compared with group C, the amount of LDH released and AR were significantly increased, Silt2 and Robo1 mRNA and protein expression was up?regulated, and no significant change was found in Robo4 mRNA and protein expression in A∕R group. Compared with group A∕R, the amount of LDH released and AR were significantly decreased in SP and H groups, and Silt2 and Robo1 mRNA and protein expression was up?regulated, and no significant change was found in Robo4 mRNA and protein expression in SP group. Compared with group SP, the amount of LDH released and AR were significantly increased, and Silt2 and Robo1 mRNA and protein expression was down?regulated in H group. Conclusion HIF?1α mediates reduction of apoptosis in cortical neurons of rats by sevoflurane preconditioning, and the mechanism is associated with Slit2∕Robo1 signaling pathway, but not with Slit2∕Robo4 signaling pathway.

Relationship between NMDA receptor and postoperative fatigue syndrome and its associated central mechanism / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the central mechanism of postoperative fatigue syndrome by detecting the expression of NMDA receptor and tryptophan metabolism.</p><p><b>METHODS</b>After being numbered according to the weight, ninety-six male SD rats were randomly divided into control group (bowel loop was flipped after laparotomy and received intraperitoneal injection of saline at a dose of 1 ml/kg), POFS model(70% of the length of small intestine was resected and received intraperitoneal injection of saline at a dose of 1 ml/kg), and NMDA antagonist groups(70% of the length of small intestine was resected and received intraperitoneal injection of MK801 at a dose of 1 ml/kg). Each group was divided into subgroups by postoperative 1, 3, 5 and 7 d, with 8 rats in each subgroup. The hippocampus was removed at each time point after open field test (OFT) to detect the mRNA expression levels of NMDA receptor 1 and kynurenine aminotransferase III((KATIII() by real-time PCR. Protein level of NMDA receptor 1 was detected by Western blot. High performance liquid chromatography (HPLC) was used to measure the concentrations of tryptophan (TRP), kynurenine (KYN) and kynurenic acid(KYNA). Ultra-structural changes of hippocampal neurons were observed by transmission electron microscopy(TEM).</p><p><b>RESULTS</b>As compared to control group, exercise score decreased(P<0.05), rest time and central panel residence time prolonged, periphery/central panel ratio increased (all P<0.05), mRNA and protein expressions of NMDA receptor 1 increased (P<0.05), mRNA expression of KAT III( decreased (P<0.05), KYN/TRP ratio and KYN/KYNA ratio decreased (all P<0.05) in POFS group on postoperative day 1 and 3. As compared to POFS group, central panel residence time and periphery/central panel ratio decreased on postoperative day 1, and mRNA and protein expressions of NMDA receptor 1 decreased on postoperative day 1 and 3 (all P<0.05) in antagonist group. TEM revealed that degenerated neuron was found in the hippocampus of POFS rats, while such damage was improved in antagonist group.</p><p><b>CONCLUSION</b>The increased expression level of NMDA receptor may play an important role in POFS. NMDA receptor antagonist MK801 may improve the POFS.</p>

Effects of Learning Environment on Learning and Memory and Microvessel Density in Hippocampus of Rats after Cerebral Infarc-tion / 中国康复理论与实践

Objective To study the effects of learning environment on learning memory and microvessel density in hippocampus of rats after cerebral infarction. Methods 80 Sprague-Dawley rats were coagulated right middle cerebral arteries electrically, and randomly divided into learning envionment (LE) and standard environment (SE) groups. They were tested with Morris Water Maze 7 d and 28 d after opera-tion. The microvessel density in hippocampus was measured with CD34 immunohistochemstry 1, 3, 7, 14 and 28 days after operation. Re-sults The achievement of Morris Water Maze was significantly better in the LE group than in the SE group (P<0.001), while the microvessel density in the hippocampus was more since 7 days after operation (P<0.05). Conclusion The learning environment can promote the angio-genesis and improve the learning and memory function recovery.

Puerarin promotes the proliferation of osteoblasts in older female patients with osteoporosis / 中国组织工程研究

BACKGROUND:In recent years, there are many animal studies and osteoblast studies on the anti-osteoporotic effects of puerarin, a kind of phytoestrogen. But few of them are reported on the effects of puerarin on osteoblasts in older patients with osteoporosis. OBJECTIVE: To observe the influence of puerarin on the proliferation ofin vitro cultured osteoblasts from older female patients with osteoporosis. METHODS:The older female patients with osteoporotic femoral neck fractures who underwent artificial femoral head replacement were included in this study. The femoral neck cancelous bone removed during the operation was colected. Primary cancelous bone osteoblasts were cultured using explant culture method. The cels were sub-cultured to the required amounts. Osteoblasts from the control group were cultured with culture medium without puerarin. Osteoblasts from the 0.01, 0.1, and 1 μmol/L puerarin groups were cultured with culture medium containing the corresponding concentrations of puerarin. After in vitro co-culture with different concentrations of puerarin for 1, 3, 5 days, the proliferation of osteoblasts was observed. RESULTS AND CONCLUSION:With the increase in the concentration of puerarin, the proliferative activity of osteoblasts constantly increased at different time points (P < 0.05). At 3 days of culture, cel absorbance value in each group reached the peak level. These results suggest that 0.01, 0.1, 1μmol/L puerarin promotes the proliferation of osteoblasts in older patients with osteoporosis in a concentration-dependent manner.

miRNA-7 inhibits proliferation of human nasopharyngeal 5-8F carcinoma cells via EGFR/PI3K/Akt pathway / 中国病理生理杂志

AIM: To investigate the relationship of microRNA-7 ( miRNA-7 ) over-expression and epidermal growth factor receptor (EGFR)/phosphatidylinositol kinase-3 (PI3K)/protein kinase B (PKB, also called Akt) pathway in human nasopharyngeal carcinoma 5-8F cells.METHODS:The 5-8F cells were transfected with miRNA-7 mimics (car-rying by Lipofectamine 2000).The expression of miRNA-7 was detected by real-time PCR.The cell proliferation was ana-lyzed by CCK-8 assay.The cell colony-forming capability was determined by cell colony formation test.The expression of EGFR/PI3K/Akt at mRNA and protein levels was examined by real-time PCR and Western blotting.RESULTS:The ex-pression level of miRNA-7 was significantly increased in 5-8F cells compared with negative control ( NC) group and control group ( P<0.01) .The proliferation of NPC 5-8F cells was decreased extremely after tansfected with the miRNA-7 mimics (P<0.01), so did the result of the cell colony-formation test.The expression of EGFR/PI3K/Akt at mRNA and protein levels was significantly down-regulated compared with NC group and control group (P<0.01).CONCLUSION:Over-ex-pression of miRNA-7 significantly inhibits the proliferation and colony-forming ability of nasopharyngeal carcinoma 5-8F cells by down-regulation of EGFR/PI3K/Akt pathway.

Pull-in small bile duct's choledochojejunostomy in rabbits / 中华肝胆外科杂志

Objective To investigate the effect of Pull in choledochojejunostomy which apllied in rabbits' Small bile duct construction on prevention of anastomosis stenosis and potential mechanisms.Methods A total of 21 rabbits were randomly assigned to three groups(n=7).Group A underwent a simple laparotomy (SL),group B biliary-enteric sutured by mucosa to mucosa Choledochojejunostomy (CJ) and group C Pull-in Choledochojejunostomy(PCJ).TBil and DBil were test in 2,4,8 weeks.The tissue of bile duct and anastomotic stoma were collected after Rabbits was killed in 8 weeks.The diameter of bile duct lumen,anastomotic stoma and the thickness of bile duct were measured respectively.Pathological changes of anastomotic stoma were observed and ki67 expression was studied by Immunohistochemical staining.Results (1)Bilirubin level was nomal in group A and C,but significantly higher in group B(P＜0.01).(2)Anastomotic stoma in group C was larger than that in group A and even more than that(completely closed) in group B.There was also statistically significant difference in the diameter of anastomotic stoma,C＞A＞B(P＜0.01).The diameter of bile duct lumen and the thickness of bile duct showed the similar results,B＞C＞A(P＜0.01).(3) As for the inflammatory studies,Group B was observed with significant infiltrated neutrophils compared with group C.Furthermore,cytokine studies showed that the expression of Ki67 index around anastomosis was significantly difference among three groups,B＞C＞A(P＜0.01).Conclusions The studies suggested that pull-in choledochojejunostomy which apllied in rabbits' small bile duct reconstruction could offer some beneficial effect in preventing anastomotic stoma stenosis.The mechanism might be through reducing the inflammatory reaction and restraining excessive hyperplasia in the area around anastomotic stoma.