Objective:To study the effect of different substrates coated on the cell survival and neurite outgrowth of spinal motor neurons (SMN) from embryonic rat cultured in vitro.Methods:The ventral spinal tissue was isolated from embryonic rats and digested into dissociated cell suspention for culture,then the cells were identified as SMN by immunohistochemistry stain.Poly-L-lysine(PLL) was dissolved into distilled water,phosphate-buffered saline solution,boric acid at 0.1 and 0.01 mol/L concentration respectively.The different substrates include PLL,collagen Ⅰ,laminin and PLL combined with laminin.Distilled water was used as control.The neuron survival numbers and the mean length of the neurites were measured and compared.Results:The cells on the PLL dissolved into boric acid at 0.01 mol/L concentration survived well.The SMNs grown on the PLL combined with laminin were in dispersed disitribution with high survival rate.Conclusion:PLL combined with laminin is the best for the study of the motor neuron including both soma and neurite.

Objective To investigate the risk factors for postoperative liver failure of patients with hepatocellular carcinoma (HCC) and bile duct tumor thrombus through a risk evaluation model.Methods The clinical data of 107 patients with HCC and bile duct tumor thrombus who received hepatic resection at the Eastern Hepatobiliary Surgery Hospital from March 2002 to February 2011 were retrospectively analyzed.All patients were divided into the non-liver failure group (98 patients) and liver failure group (9 patients).Risk factors associated with liver failure were analyzed and a risk evaluation model was established.All data were analyzed using the bivariate regression model,and factors with significance were further analyzed using the multivariate regression model.Results Of the 107 patients,105 received hepatic resection + choledochotomy + thrombectomy and 2 received hepatic resection + extrahepatic bile duct resection + cholangiojejunostomy.The operation time was 2.0-5.5 hours,and the intraoperative blood loss was 200-3500 ml.In the non-liver failure group,5 patients had pleural and peritoneal effusion,3 had biliary bleeding,2 had incisional infection,1 had biliary infection,1 had bile leakage,1 had stress-induced ulcer of upper digestive tract and 1 had thoracic epidural hematoma.The bleeding of the patients with thoracic epidural hematoma was stopped after thoracic spinal decompression,but subsequent paraplegia occurred.In the liver failure group,2 patients died of postoperative acute liver failure,and 7 patients died of postoperative subacute liver failure (death caused by tumor recurrence or medicine was excluded).The results of univariate analysis showed that preoperative total bilirubin,albumin,pre-albumin,albumin/globulin ratio,distribution of tumor thrombus,operative blood loss and ratio of postoperative residual liver volume to the total liver volume were correlated with the postoperative liver failure in patients with HCC and bile duct tumor thrombus (OR =3.017,0.191,0.248,2.681,9.048,4.759,13.714,P ＜ 0.05).The results of multivariate analysis showed that preoperative total bilirubin ＞ 256.5 μmol/L,albumin/globulin ratio ≤ 1.3 and postoperative residual liver volume ＜ 50％ were the independent risk factors of postoperative liver failure (OR =5.537,11.107,172.450,P ＜ 0.05).The risk evaluation model was Z =1.77 × preoperative total bilirubin + 2.408 × preoperative albumin/globulin ratio + 5.150 × ratio of postoperative residual liver volume to the total liver volume-17.288.The risk of postoperative liver failure increased as the increase of Z value.The risk of postoperative liver failure ＞ 50％ when the Z value ＞ 0.Conclusions Preoperative total bilirubin ＞ 256.5μmol/L,albumin/globulin ratio ≤ 1.3 and postoperative residual liver volume ＜ 50％ were the independent risk factors of postoperative liver failure.Risk evaluation model is helpful in screening the risk factors so as to decrease the incidence of postoperative liver failure.

Objective To investigate the effect of heme oxygenase-1 ( HO-1 ) on the acute radiation-induced skin injury by gene transfer.Methods Thirty-three male SD rats were randomly divided into three groups as PBS-injected group,Ad-EGFP-injeeted group and Ad-HO-1-injected group ( n =11 ).In each group,three rats were used for determining the expression of target gene and the other rats were irradiated on the buttock skin with 40 Gy electron beam generated by a linear accelerator.Immediately after irradiation,rats were administered with a subcutaneous injection of PBS,Ad-EGFP or Ad-HO-1,respectively.Subsequently,the skin reactions were measured twice a week using the semi-quantitative skin injury scale.Results The strong positive expression of HO-1 was observed in subcutaneous dermal tissue after injection of Ad-HO-1.Compared to the PBS-injected group or the Ad-EGFP-injected group,a significant mitigation of skin injury was observed in Ad-HO-1-injected mice 14 d after irradiation (q =0.000-0.030,P ＜ 0.05 ).Conclusions HO-1 could significantly mitigate radiation-induced acute skin injury and Ad-HO-1 could be used to treat radiation-induced skin injury.

Objective To investigate the roles of somatostatin(SS)positive intemeurons in the development and compensation of temporal lobe epilepsy.Methods Piloearpine-induced epilepsy rat model was established.Immunohistochemistry method was used to detect number changes and axonal sprouting of SS positive intemeurons in different domains of the hippocampus at difierent time points.Degeneration of SS positive interneurons and their neurophils were detected by the double immunofluorescence staining with SS and Fluoro-Jade B(FJB)at 7 and 60 days after status epilepticus (SE).Results In the exoerimental rat group,the number of SS positive neurons decreased in each hippocampal domain,and it reached the lowest at 7 days post-SE(There were 11.1±3.3 in hilus,2.8±0.9 in CA1region and 1.8±0.7 in CA1region,t=13.519,9.644 and 8.808,all P<0.01).In chronic phase,the number of SS neurons gradually recovered,and exceeded the control group in CA1 area at 60 days post-SE(12.8±1.5 vs 8.8±1.3,t=-4.506,P<0.01),however,the number of SS neurons in the hilus(25.5±4.6)and CA1 area(4.8±0.8)remained significantly less than normal levels(t value were 4.691 and 3.953.both P<0.01).Increased SS positive fibers were found in the lacunosum-molecular (1m)layer and outer molecular layer of dentate gyrus after 30 days post-SE,and numerous SS positive fibers were seen threnghout the layers of area CA1 at 60 days post-SE.Double immunofluuorescence revealed that a few SS positive interneurons and fibers were also labeled by FJB in area CA1 at 7 days post-SE and in CA domain/hilus at 60 days post-SE.Conclusions SS intemeurons loss plays an important role in the development of temporal lobe epilepsy.The loss is partially caIlsed by the degeneration and death of neurons;SS positive neurophils increase within area CA1 in chronic phase may play a significant role in the generation and compensation of temporal lobe epilepsy.

Objective To investigate the expression of XIAP, Smac, HtrA2 and XAF1 in the hippocampus following SE in rats and to explore the pathophysiological mechanisms of expression of XIAP and its negative regulators after SE. Methods The lithium-pilocapine model of status epilepticus was established in SD rat. XIAP, Smac, HtrA2, XAF1 and activated caspase-3 protein were examined using immunohistochemistry. Western blot was used to detect the protein levels of XIAP, Smac, HtrA2 and activated easpase-3. Results XIAP immunoreactivity diffusely distributed within the neuron after SE. Compared with the control group, the expression of CA3 XIAP protein in the SE group was increased gradually since 2 hours (0.5503±0.0172 vs 0.1507±0.0165, t=115.87, P<0.01), peaking at 8 hours (0.6221±0.0238 vs 0.1507±0.0165, t=136.69, P<0.01). The expression of CA3 Smac, HtrA2, XAF1 and activated caspase-3 protein were increased generally following SE. Western blot analysis showed a significant increase in Stoat, HtrA2, activated caspase-3 protein levels from 2 to 72 hours following SE, but no significant differences were seen in XIAP protein levels between the control group and the SE group. Conclusions The XIAP, Smac, HtrA2 and XAF1 are involved in the regulation of neuronal apoptosis and implicated in pathophysiological mechanisms of neuronal damage after SE.

Objective To quantitatively assess the cardiopulmonary exercise function of spinal cord injury (SCI) patients and observe the effect of aerobic exercise on cardiopulmonary function, motor function and activities of daily living. Methods From December, 2014 to June, 2016, 34 incomplete SCI patients (ASIA C and D) and 23 healthy controls received cardiopulmonary exercise test (CPET). SCI pa-tients were randomly divided into conventional rehabilitation group (n=17) and aerobic exercise group (n=17). The aerobic exercise group received aerobic exercise for four weeks. They were assessed with CPET, motor and sensory function, walking index for spinal cord injury II (WISCI II) and spinal cord independence measure (SCIM) before and four weeks after training. Results Oxygen uptake (VO2)peak, anaerobic threshold (AT), metabolic equivalent of energy (METpeak), VO2/heart rate (HR)peak, respiratory exchange rate (RER)peak, minute ventilation (VE)peak, work rate (WR)peak and systolic blood pressure (SBP)peak were lower in the patients than in the controls (t>2.714, P<0.05). VO2peak、AT、METpeak、VO2/HRpeak、WRpeak increased in the aerobic exercise group after training (t>2.431, P<0.05). METpeak and WRpeak improved in the conventional rehabilitation group after training (t>3.282, P<0.01). The scores of motor in ASIA and SCIM improved in both groups after training (t>2.985, P<0.05). Conclusion The cardiopulmonary function decreased in incomplete SCI patients, which could be improved by moderate intensity aerobic exercise.

AIM: G protein-coupled inwardly rectifier potassium (GIRK) channel are distributed widely in mammalian brain. In CNS, GIRK 1/2 seems to be the predominant heterotetramers which play a pivot role in the regulation of the excitability of neurons and may contribute to the resting potential by leading to a hyperpolarization of membrane potential and reduction of the action potential frequency. In the context, the Weaver mouse is the first neurological abnormality directly linked to a genetic point mutation in the GIRK2 protein which includes spontaneous seizure. GIRK2 knock out mice showed normal development but more susceptible than normal mice to seizure induced by GABA antagonist. Here, we report that the mRNA and protein expression of GIRK subunit 2 is altered in kainic acid(KA)-induced epileptic rat hippocampus. METHODS: Rats were injected with kainate 14 mg/kg intraperitoneally to establish an acute and chronic temporal lobe epilepsy model. At chronic spontaneous seizure stage, by using of in situ hybridization, immunocytochemistry and Western blotting, the GIRK 1,2 mRNA and protein were analyzed quantitatively in the dentate gyreus, CA1, CA3 regions of hippocampus. RESULTS: GIRK1,2 mRNA and proteins were expressed abundantly in all regions of hippocampus. KA induced seizures and caused a significant increase in GIRK2 mRNA abundance and immunoreacitivity; only GIRK1 mRNA was increased significantly, but no difference was found by Western blotting protocol. CONCLUSION: GIRK1,2 mRNA and protein expression in the hippocampus of epileptic rat brain is up-regulated, which may be an adaptive response to over-excitability of neuron networks and prevent the over-excitability spread in hippocampus (DG-CA3-CA1). [

Objective:To screen for the bioactive marine organisms and to study the active constituents from them.Methods:Bioactivities of marine organisms extracts were screened by Pyricularia oryzae bioassay method and the constituents were isolated and identified by a combination of multi chromatography and spectral analysis.Results:Twenty ethanolic extracts and 5 aqueous extracts of the tested marine organisms showed activities causing morphological abnormality of P.oryzae mycelia.Fifty six compounds were obtained and identified from Sargassum carpophyllum,Ishige okamurai and Natarchus leachii freeri ,among which 28 compounds showed activities against P.oryzae and exhibited cytotoxicities on various cultured tumor cell lines,eight compounds showed inhibition effects on Fonsecaea pedrosoi in vitro .Conclusion:The screening data and bioactivity evaluation suggest the P.oryzae bioassay is suitable for preliminary screening of bioactive agents from marine organisms.

OBJECTIVE: To evaluate the quality of reports published in recent 10 years in China about quantitative analysis of syndrome differentiation for diabetes mellitus (DM) in order to explore the methodological problems in these reports and find possible solutions. METHODS: The main medical literature databases in China were searched. Thirty-one articles were included and evaluated by the principles of clinical epidemiology. RESULTS: There were many mistakes and deficiencies in these articles, such as clinical trial designs, diagnosis criteria for DM, standards of syndrome differentiation of DM, case inclusive and exclusive criteria, sample size and estimation, data comparability and statistical methods. CONCLUSION: It is necessary and important to improve the quality of reports concerning quantitative analysis of syndrome differentiation of DM in light of the principles of clinical epidemiology.

Objective:To investigate the value of atropine administration before tracheal intubation under general anesthesia in the elderly patients undergoing endobronchial ultrasound-guided transbronchial needle aspiration(EBUS-TBNA)biopsy.Methods:This was a case-control study.A total of 87 elderly hospitalized patients receiving chest enhanced CT test suggesting the risk of lung cancer with enlargement of mediastinal lymph nodes in 7 regions were scheduled to undergo endobronchial ultrasound-guided transbronchial needle aspiration(EBUS-TBNA)biopsy under general anesthesia.The patients were randomly divided into two groups by flipping a coin: the atropine group(n=40)and the control group(n=47). The indicators for evaluating the application values of atropine included preoperative, intra-operative and postoperative systolic blood pressure(SBP), diastolic blood pressure(DBP), heart rate, oropharyngeal and airway secretion volume, oxygen saturation, operation time, positive diagnosis rate and postoperative adverse reactions.Results:SBP and DBP were lower in the atropine group than in the control group before endotracheal intubation(131.7±15.3 mmHg vs.140.7±13.7 mmHg, 79.1±7.6 mmHg vs.85.6±7.4 mmHg, t=2.885 and 4.061, P<0.05). There was no significant difference in SBP and DBP 10 min after endoscopic operation and 10 min after tracheal extubation between the atropine group versus the control group(SBP: 109.1±11.2 mmHg vs.105.0±12.2 mmHg, 136.9±23.0 mmHg vs.129.9±11.2 mmHg, DBP: 66.9±7.5 mmHg vs.68.0±8.3 mmHg, 77.6±10.9 mmHg vs.78.5±6.4 mmHg, t=-1.617, 0.687, -1.751 and 0.448, P>0.05). There was no significant difference in HR between the two groups before endotracheal intubation( t=1.416, P>0.05), while HR was higher in the atropine group than in the control group 10 min after endoscopic operation and 10 min after tracheal extubation( t=-3.323 and -2.181, P<0.01 and P<0.05). The change rates of SBP and DBP were lower in the atropine group than in the control group 10 min after endoscopic operation and 10 min after tracheal extubation( t=7.947, -6.962, -3.187 and -3.232, P<0.01). The change rate of HR was lower in the atropine group 10 min after endoscopic operation and was higher 10 min after tracheal extubation than in the control group( t=-6.467 and -4.131, P<0.01). There were significant differences in the volume of oropharyngeal and airway secretions and fingertip oxygen saturation between the two groups before endotracheal intubation and 10 min after tracheal extubation( t=-2.334, 2.759, -3.314 and -2.767, P<0.01). The endoscopic operation time was less in the atropine group than in the control group with no statistically significant difference[(25.9±5.7)min vs.(26.4±4.7)min, t=0.391, P>0.05]. There was no significant difference in postoperative adverse reactions between the atropine group versus the control group(34 patients or 85.0% vs.43 patients or 91.5%, χ2=1.247, P>0.05). Conclusions:The application of atropine before tracheal intubation under general anesthesia is beneficial to stabilizing the intraoperative blood pressure and heart rate, and can reduce the production of postoperative oropharyngeal and airway secretions in elderly patients undergoing endobronchial ultrasound-guided transbronchial needle aspiration biopsy.