Objective To study the effect of ketamine on L-type calcium currents (ICa-L) in guinea pig ventricular myocytes. Methods Adult guinea pigs of both sexes were anesthetized with pentobarbital. The hearts were immediately removed and ventricular myocytes were prepared by the technique described by Liu et al. The whole-cell patch clamp technique was used to study the ICa-L in isolated guinea pig ventricular myocytes. The changes in ICa-L produced by ketamine 100 ?mol?L-1 with different holding potentials or by different concentrations of ketamine with holding potential of + 10 mV were analyzed. Results Ketamine dose-dependently inhibited ICa-L evoked by a voltage step from a holding potential of - 40 mV to + 10 mV. The 4 concentrations of ketamine (100, 500, 1 000, 5 000 ?mol?L-1) reduced 1Ca-L by 28.7%?5.7% , 34.7%?1.4%, 58.7%?6.4% and 81.7%?6.7% respectively, with a mean IC50 concentration of 926.6 ?mol?L-1 . When the cells were exposed to ketamine 100 ?mol?L-1, the steady-state activation curve was not significantly affected, while the steady-state inactivation curve was shifted to more negative potentials.V1/2 decreased from ( - 14.8?0.8 ) mV to ( - 19.6?0.7) mV (P0.05) in control and drug-affected cells respectively. Ketamine slowed the rate of recovery from inactivation. Conclusion Ketamine can inhibit ICa-L in guinea pig ventricular myocytes in a concentration-dependent manner. This inhibitory effect of ketamine may explain its negative inotropic effect. Ketamine inhibits L-type calcium channel in its inactivated state.

Objective To determine the effect of cardiomyopeptidin on transient outward potassium current(I_(to) of rat ventricular myocytes and its action mechanism on the ion channels of myocardium.Methods Single ventricular myocytes of rats were obtained by enzymatic dissociation.The whole-cell patch-clamp recording technique was used to record the change of transient outward potassium current(I_(to) by different dosages of cardiomyopeptidin.Results Cardiomyopeptidin decreased I_(to) in a dose-dependent manner.Cardiomyopeptidin in dose of 10,50,100,250 and(500 mg/L) decreased I_(to %) by 4,13,22,32 and 38 respectively.Cardiomyopeptidin 50 mg/L moved the current density-voltage curve of I_(to) down,but the shape of the curve had no changes.Cardiomyopeptidin 50 mg/L did not change the steady state activation curve of I_(to).Conclusions Cardiomyopeptidin decreases the I_(to) of rat ventricular myocytes,which might be one of the mechanisms of its antiarrhythmic effect.

Objective To investigate influence of ulinastation in storage period on apoptosis of suspended erythrocyte.Methods RBCs were treated with saline (control group) and different doses of ulinastatin (5,000, 10,000 and 50,000 U/mL in group C1, C2 and C3, respectively).samples were detected when stored at 0,7,14,21,28,35 d,respectively.Indicators of corpuscular volume,phosphatidylserine extroversion rate and intracellular Ca2 +concentration were analyzed by flow cytometer.Results The phosphatidylserine (PS)-exposure levels of 4 groups started to increase on 14 day(P<0.05). Cells of the control group, group C1 and C2 began to shrink remarkably on day 21, while that of Group C3 on 28 day.The intracellular Ca2 +levels of the control group and group C1 started to increase significantly on day 35, (t=16.33,t=14.66,P<0.05).one Ca2 +levels of group C1,C2 and C3 increased on day 14.From 21 to 35 day, the intracellular Ca2 +levels of group C2 and C3 were no significant compared with control group.Conclusion During the storage period, suspended erythrocyteapoptosis increase with time prolonged, adding suitable amount of ulinastatin in stock solution can inhibit apoptosis in damage at some level.

AIM: To study and evaluate the changes of two main kinds of voltage-gated K+ currents in human atrial fibrillation (AF) and to discuss the role of these changes in the atrial electrical remodeling (AER) caused by AF. METHODS: Specimens of human atrial appendage were obtained from 36 RHD patients (18 with chronic AF and 18 without AF). Single atrial myocytes were acutely dissociated by tissue chunk enzymatic digestion. I_~to1 and I_~Kur in the two groups were measured respectively with the patch-clamp technique in a whole-cell configuration and the I-V curves were compared. RESULTS: I_~to1 and I_~Kur amplitudes in AF groups were significantly reduced and the current densities of both I_~to1 and I_~Kur in AF patients were lower than those in NAF patients. CONCLUSION: The reduction of I_~to1 and I_~Kur may be related to changes in atrial conduction, refractory period and may constitute two main parts of the major mechanisms in the AER of chronic AF. Whether exists a relation between changes of the above K+ currents and that of other ionic currents and the AF initiation and perpetuation deserves further investigation. [

Objective To explore the clinical features of hemophagocytic syndrome in children and the signifi﹣cance of gene detection. Methods TWenty－tWo pediatric patients diagnosed as hemophagocytic syndrome since 2004 clinical and laboratory criteria Were enrolled in Children＇s Hospital Affiliated to Zhengzhou University from January 2014 to January 2016. The clinical data of patients Were analyzed,and the genes associated With hemophagocytic syn﹣drome Were detected. The clinical biochemical indicators Were compared betWeen mutation group and non －mutation group. Results TWenty－tWo cases of patients(3 months to 12 years)Were enrolled,including 10 males and 12 fe﹣males,and the proportion of children over 5 years old accounted for the highest proportion,accounting for 50%,and all of them had fever,liver,spleen and lymph node enlargement. The main test results Were as folloWs:peripheral blood cells decreased in 6 cases( 27. 27%),hemophagocytic phenomena presented in bone marroW smears in 12 cases (54. 55%),abnormal liver function in 18 cases(81. 82%),and loW serum albumin in 22 cases(100. 00%). High serum ferritin levels Were detected in 20 cases(90. 91%);the detection of natural killer(NK)cell activity shoWed nor﹣mal activity( active ＞ 15%) in 7 cases( 31. 82%),and decreased activity( activity ≤ 15%) in 15 cases (68. 18%). The genes associated With hemophagocytic syndrome Were detected in 22 cases of patients,and 12 of them Were associated With mutations related to hemophagocytic syndrome,accounting for 54. 55%. LYST,ITK and UNC13D gene Were common. There Was no statistical difference in Which ages of onset,symptoms of the nervous system,and labo﹣ratory data of leukocyte count,red blood cell count,hemoglobin,platelet count,NK cell activities,prognosis,hemophago﹣cytic phenomena shoWed in bone marroW smears,alanine aminotransferase,albumin,triglyceride,ferritin and fibrinogen betWeen mutation group and non－mutation group(all P＞0. 05). Conclusions Pediatric hemophagocytic syndrome is mostly accompanied by fever,liver,spleen and lymph node enlargement,and most of them are accompanied by gene mu﹣tations. LYST,ITK and UNC13D gene are commonly seen. But there is no significant correlation betWeen gene mutation and general condition,biochemical index and severity of the disease.

Objective To investigate the preventive effect and mechanisms of Naoshuantong capsule in stroke induced by artificial cold wave in hypertensive rats.Methods A total of 130 SD rats were randomly divided into a sham operation group (n =30),a model control group (n =50) and a Naoshuantong treatment group (n=50;intragastric administration of Naoshuantong,0.5 g/kg,2/d for 7 days).Renovascular hypertensive rats model were established by two-kidney,two clip method.At 13th week after operation,rats were exposed to artificial cold wave for 3 days (12 h light of 22 ℃ and 12 h dark of 4 ℃,3 cycles).The brain tissue samples were extracted at the end of the experiment.Differential protein proteomic techniques were used for the identification,functional classification and preliminary analysis of the differentially expressed protein spots,and Western blot was used for the validation of some key proteins.Results There was no occurrence of stroke in the sham operation group,and the incidence of stroke in the model control group (36.00％,18/50) was significantly higher than that in the Naoshuantong treatment group (18.00％,9 / 50;x2 =4.110,P =0.043).With the two-dimensional gel electrophoresis analysis,6 different proteins were identified from 14 protein spots.Among them,the up-regulated superoxide dismutase 2 (SOD2) and the down-regulated B-cell lymphoma 10 (Bcl-10) were found to be at the central location of protein interactions,which has been verified by Western blot.Conclusion Naoshuantong can reduce the occurrence of stroke induced by artificial cold wave in renovascular hypertensive rats.SOD2 up-regulating and Bcl-10 downregulating may be involved in the mechanisms of of Naoshuantong in the prevention of cold wave-induced stroke in hypertensive rats.

Objective:To investigate the expression of Per2 mRNA, HDAC1 mRNA and E-cadherin mRNA in esophageal cancer cells and their significance.Methods:The experimental study was conducted. Human normal esophageal epithelial cells as the control group and human esophageal cancer cell line KYSE-150 cells as the experimental group were cultured in vitro to logarithmic growth stage. Observation indicators: (1) the proliferation of cells; (2) the migration and invasion of cells; (3) the expression of Per2 mRNA, HDAC1 mRNA, and E-cadherin mRNA in cells of initial physiological state; (4) the expression of Per2 mRNA, HDAC1 mRNA, and E-cadherin mRNA after cells were treated with Per2-agonists or inhibitors; (5) the expression of Per2 mRNA and E-cadherin mRNA after cells were treated with HDAC1 inhibitors. Measurement data with normal distribution were represented as Mean± SD, the t test was used for comparison within groups and the t test or ANCOVA were used for comparison between groups. Results:(1) The proliferation of cells: the cell proliferation of the experimental group and control group were 0.90%±0.14% and 0.52%±0.08%, with a significant difference between the two groups ( t=5.166, P<0.05). (2) The migration and invasion of cells: the numbers of cell migration and invasion for the experimental group were 173±41 and 86±27, versus 50±15 and 21±9 for the control group, with significant differences between the two groups ( t=6.274, 5.153, P<0.05). (3) The expression of Per2 mRNA, HDAC1 mRNA, and E-cadherin mRNA in cells of initial physiological state: the expression of Per2 mRNA, HDAC1 mRNA, and E-cadherin mRNA in cells of initial physiological state for the experimental group was 11.7±2.7, 20.4±6.6, and 12.4±2.5, respectively, versus 2.4±0.5, 8.5±2.2, and 27.3±4.5 for the control group, with significant differences between the two groups ( t=5.782, 2.982, -5.034, P<0.05). (4) The expression of Per2 mRNA, HDAC1 mRNA, and E-cadherin mRNA after cells were treated with Per2-agonists or inhibitors: after cells were treated with Per2-agonists, the expression of Per2 mRNA, HDAC1 mRNA, and E-cadherin mRNA were 13.1±2.2, 22.4±6.2, 16.6±4.2 for the experimental group, and 9.9±3.1, 18.4±5.6, 15.3±2.3 for the control group, respectively. There was no significant difference in the expression of Per2 mRNA, HDAC1 mRNA, or E-cadherin mRNA of the experimental group between cells being treated with and without Per2-agonists ( t=-4.300, 10.087, -4.187, P>0.05). There were significant differences in the expression of Per2 mRNA, HDAC1 mRNA, and E-cadherin mRNA of the control group between cells being treated with and without Per2-agonists ( t=-4.846, 3.501, 9.294, P<0.05). There was no significant difference in the expression of Per2 mRNA or E-cadherin mRNA between the experimental group and control group after cells were treated with Per2-agonists ( F=1.000, 7.582, P>0.05), while there was a significant difference in the expression of HDAC1 mRNA between the two groups ( F=1.724, P<0.05). After cells were treated with Per2-inhibitors, the expression of Per2 mRNA, HDAC1 mRNA, and E-cadherin mRNA were 4.1±1.7, 7.5±2.2, 22.8±4.2 for the experimental group, and 3.1±0.9, 9.3±3.2, 28.4±5.8 for the control group, respectively. There were significant differences in the expression of Per2 mRNA, HDAC1 mRNA, and E-cadherin mRNA of the experimental group between cells being treated with and without Per2-inhibitors ( t=12.124, 5.105, -10.245, P<0.05). There was no significant difference in the expression of Per2 mRNA, HDAC1 mRNA, or E-cadherin mRNA of the control group between cells being treated with and without Per2-inhibitors ( t=-2.815, 1.568, -1.439, P>0.05). There were significant differences in the expression of Per2 mRNA and E-cadherin mRNA after cells were treated with Per2-inhibitors between the experimental group and control group ( F=22.965, 82.134, P<0.05), while there was no significant difference in the expressions of HDAC1 mRNA between the two groups ( F=6.416, P>0.05). (5) The expression of Per2 mRNA and E-cadherin mRNA after cells were treated with HDAC1 inhibitors: after cells were treated with HDAC1 inhibitors, the expression of Per2 mRNA and E-cadherin mRNA were 13.4±3.5, 24.2±3.4 for the experimental group, and 3.1±1.2, 26.8±5.2 for the control group, respectively. There was no significant difference in the expression of Per2 mRNA of the experimental group between cells being treated with and without HDAC1-inhibitors ( t=-3.959, P>0.05). There was a significant difference in the expression of E-cadherin mRNA of the experimental group between cells being treated with and without HDAC1-inhibitors ( t=-21.977, P<0.05). There was no significant difference in the expression of Per2 mRNA or E-cadherin mRNA of the control group between cells being treated with and without HDAC1-inhibitors ( t=-1.440, 1.058, P>0.05). After cells were treated with HDAC1-inhibitors, there was no significant difference in the expressions of Per2 mRNA between the experimental group and control group ( F=2.004, P>0.05), while there was a significant difference in the expression of E-cadherin mRNA between the two groups ( F=325.800, P<0.05). Conclusions:Human esophageal cancer cells have an elevated expression of Per2 mRNA and HDAC1 mRNA, and a reduced expression of E-cadherin mRNA. The overexpression of Per2 mRNA may activate the expression of downstream targeting protein HDAC1, and inhibit the expression of cell surface E-cadherin mRNA.

Objective To summarize risk factors for clinical outcomes in heart transplantation patients, evaluate the characters of Chinese patients by comparing with international data, and introduce new clinical strategies. Methods We performed 200 heart transplantations from Jun. 2004 to May 2010. The clinical information was recorded and all patients were followed up. By analyzing 160 patients with a follow-up period of more than one year, we summarized clinical outcomes and risk factors of early and late results of heart transplant patients. Results Of 160 patients, 8. 1 % received postoperative extracorporeal membrane oxygenation (ECMO) support and 10% continuous renal replacement therapy. In 550 cases/times of endomyocardial biopsies, the incidence of rejection with grades more than Ⅱ (concluding grade Ⅱ ) was 14. 9%. In-hospital mortality was 3. 8%. Smoking,preoperative diastolic pulmonary arterial pressure, PAWP, total serum protein level and albumin level were risk factors of peri-operative mortality, and preoperative diastolic pulmonary arterial pressure,primary heart diseases, pulmonary hypertension and implantations of ICD, MCS and ECMO were risk factors of late mortality. Postoperatively, 1-, 3- and 5-year survival rate was 94. 4%, 91.9% and 88. 8%, respectively. Compared with UNOS data, the rate of primary heart diseases, pulmonary hypertension, and implantation of ICD, MCS and ECMO were different, and the long-term survival rate of 160 patients was higher than that reported by ISHLT. Conclusion The risk factors of mortality of Chinese heart transplant patients are different with their counterparts from western countries. Our corresponding peri-operative treatments and clinical strategies have produced satisfactory clinical outcomes.

Lipotoxicity,caused by intracellular lipid accumulation,accelerates the degenerative process of cellular senescence,which has implications in cancer development and therapy.Previously,camitine palmi-toyltransferase 1C (CPT1C),a mitochondrial enzyme that catalyzes carnitinylation of fatty acids,was found to be a critical regulator of cancer cell senescence.However,whether loss of CPT1C could induce senescence as a result of lipotoxicity remains unknown.An LC/MS-based lipidomic analysis of PANC-1,MDA-MB-231,HCT-116 and A549 cancer cells was conducted after siRNA depletion of CPT1C.Cellular lipotoxicity was further confirmed by lipotoxicity assays.Significant changes were found in the lipidome of CPT1 C-depleted cells,including major alterations in fatty acid,diacylglycerol,triacylglycerol,oxidative lipids,cardiolipin,phosphatidylglycerol,phosphatidylcholine/phosphatidylethanolamine ratio and sphingomyelin.This was coincident with changes in expressions of mRNAs involved in lipogenesis.Histological and biochemical analyses revealed higher lipid accumulation and increased malondialde-hyde and reactive oxygen species,signatures of lipid peroxidation and oxidative stress.Reduction of ATP synthesis,loss of mitochondrial transmembrane potential and down-regulation of expression of mito-chondriogenesis gene mRNAs indicated mitochondrial dysfunction induced by lipotoxicity,which could further result in cellular senescence.Taken together,this study demonstrated CPT1C plays a critical role in the regulation of cancer cell lipotoxicity and cell senescence,suggesting that inhibition of CPT1C may serve as a new therapeutic strategy through induction of tumor lipotoxicity and senescence.

Objective To evaluate the clinical value of 3D reconstruction in the single utility-port thoracoscopic segmentectomy of early stage NSCLC by propensity score matching (PSM). Methods We retrospectively analyzed clinical data of 150 early stage NSCLC patients undergoing single utility-port thoracoscopic segmentectomy. The patients were divided into reconstruction group (n=58) and non-reconstruction group (n=92) according to 3D reconstruction. PSM was performed on two groups to compare perioperative outcomes. Results Procedures were successfully completed on all patients, without perioperative death. In each group, 43 patients were successfully matched after PSM on the basis of 8 confounding factors, age, gender, smoking status, BMI, maximum tumor diameter on CT, tumor location, % FEV1 and type of planned segmentectomy. After PSM, in complex segmentectomy, the patients in the reconstruction group had shorter operation time (155.77±30.17 vs. 212.94±66.49min, P < 0.001) and less blood loss (46.00±25.94 vs. 88.79±68.36ml, P=0.002), compared with the non- reconstruction group. Conclusion Preoperative 3D reconstruction could help improve the efficiency of single utility-port thoracoscopic surgery for complex segmentectomy and reduce intraoperative bleeding.