Objective To evaluate the effect of sevoflurane preconditioning on autophagy during ischemiareperfusion (I/R) injury to isolated rat hearts and the role of PI3K/Akt signaling pathway.Methods Healthy adult male Wistar rats,aged 6-8 weeks,weighing 250-280 g,were anesthetized.Their hearts were excised and perfused in a Langendorff apparatus.Eighty-four isolated rat hearts with I/R injury were randomly divided into 7 groups (n =12 each):normal control group (NC group),I/R group,sevoflurane preconditioning group (S + I/R group),normal control plus wortmannin group (NC + W group),I/R plus wortmannin group (I/R + W group),sevoflurane preconditioning plus wortmannin group (S + I/R + W group),and solvent group (S + I/R + D group).In NC group,the hearts were continuously perfused with K-H solution for 180 min.In I/R group,the hearts were perfused with K-H solution for 30 min,and then subjected to ischemia for 30 min followed by 120 min of reperfusion.In S+ I/R group,the hearts were perfused with K-H solution for 10 min,and then with K-H solution saturated with 2.5％ sevoflurane for 15 min,followed by 5 min washout with K-H solution before ischemia.In NC + W group,wortmannin (PI3K inhibitor) 15 μg/kg was injected intraperitoneally at 30 min before chest opening,and the other procedures were similar to those previously described in group NC.In I/R + W group,wortmarnin 15 μg/kg was injected intraperitoneally at 30 min before chest opening,and the other procedures were similar to those previously described in group I/R.In S + I/R + W group,wortmannin 15 μg/kg was injected intraperitoneally at 30 min before chest opening,and the other procedures were similar to those previously described in S + I/R group.In S + I/R + D group,dimethyl sulfoxide 1.5 ml/kg was injected intraperitoneally at 30 min before chest opening,and the other procedures were similar to those previously described in group S + I/R.The HR,± dp/dtmax,left ventricular developed pressure (LVDP) and left ventricular end-diastolic pressure (LVEDP) were recorded at the end of equilibration and reperfusion.At the end of reperfusion,coronary effluent was collected to detect lactate dehydrogenase (LDH) activity,and myocardial specimens were obtained to calculate the percentage of myocardial infract size.The expression of autophagy marker LC3-Ⅱ and Akt,phosphor-Akt (p-Akt),mammalian target of rapamycin (mTOR),and phosphor-mTOR (p-mTOR) was determined by Western blot.Results Compared with NC group,no significant change was found in the parameters of hemodynamics in NC + W group,and HR,± dp/dtmax and LVDP were significantly decreased,LVEDP,myocardial infract size,and LDH activity were increased,LC3-Ⅱ expression was up-regulated,and the expression of p-Akt and p-mTOR was down-regulated in the other groups.Compared with group I/R,HR,± dp/dtmax,and LVDP were significantly increased,LVEDP,myocardial infract size,and LDH activity were decreased,LC3-Ⅱ expression was downregulated,and the expression of p-Akt and p-mTOR was up-regulated in S + I/R and S + I/R + D groups,and no significant change was found in each parameter in S+ I/R+ W group.Compared with S + I/R group,HR,± dp/ dtmax and LVDP were significantly decreased,LVEDP,myocardial infract size and LDH activity were increased,LC3-Ⅱ expression was up-regulated,and the expression of p-Akt and p-mTOR was down-regulated in S + I/R + W group,and no significant change was found in each parameter in S + I/R + D group.Conclusion Sevoflurane preconditioning can decrease autophagy of myocardial cells during I/R through activating PI3K/Akt signaling pathway and enhancing mTOR activity in the downstream,thus protecting isolated rat hearts against I/R injury.

Atropine (Atr) ip 5 - 10mg/kg elevated the threshold dose of ouabain ( 10?g/min, iv ) to induce ventricular premature beats, ventricular tachycardia , ventricular fibrillation and cardiac arrest in guinea-pigs. Atr 2 - 4 mg/kg iv shortened the duration of arrhythmia elicited by Adr ( 40?g/kg, iv ) in conscious rabbits, & delayed the onset of arrhythmia evoked by AC ( 20?g/kg, iv ) or BaCl2 ( 2mg/kg, iv ) in anesthetized rats. It also decreased the incidence of arrhythmia induced by CaCl2 ( 102mg/kg, iv ) in rats. In mice, Atr ( 5~10mg/kg, ip)decreased the incidence of ventricular fibrillation by chloroform and artrial fibrill-ation ( or flutter ) by Ach-CaCl2.

AIM: To explore the effect of protein kinase C-?(PKC-?) in U937 cell line inhibited by short hairpin RNA(shRNA) on the transcription of vascular endothelial growth factor(VEGF) mRNA induced by stromal cell-derived factor-1(SDF-1).METHODS: 64 bp reverse repeated motifs of PKC-? target sequence were synthesized and inserted into the plasmid to construct the plasmid expressing shRNA-PKC-?(pSIRENp) and the pSIRENp plasmid was transfected into U937 cell line.The expression of PKC-? and VEGF mRNA was detected by RT-PCR.RESULTS: The recombinant plasmid pSIRENp was successfully constructed and it nearly completely suppressed the PKC-? expression in U937 cell line.After transfection,both basical and VEGF mRNA induced by SDF-1 significantly reduced compared to control.CONCLUSION: The results shows that the short hairpin RNA of PKC? efficiently reduces its expression in U937 cells and PKC-? may be involved in the regulation of VEGF mRNA expression.

Objective: To develop the matrix-matched calibration curve correction-inductively coupled plasma mass spectrometry (ICP-MS) for the determination of lead in blood. Methods: Whole blood samples and blank whole blood were pretreated by direct dilution with a solution of 0.5% nitric acid and 0.01% TritonX-100 to obtain whole blood sample solutions and matrix-matched solvents at a 10-fold dilution. The mass concentration of lead was determined by using an ICP-MS instrument in He mode. 175Lu was added online as an internal standard. The standard working curve was calibrated with the matrix-matched solvent, and the mass concentration of lead in the whole blood samples was calculated based on the standard working curve. Recovery tests were performed on whole blood blind samples by spiking, and the relative standard deviation and average recovery rate were calculated. The accuracy and precision of this method were assessed by comparing it with the method recommended in the national standard in detection of lead in three types of bovine blood lead standard materials. Results: Good linearity was shown for lead at 0.5 to 100.0 μg/L, with a correlation coefficient of 1.000. The detection limit of lead was 0.4 μg/L, and the quantitation limit was 1.3 μg/L. The relative standard deviations were 0.65% and 1.10%. The average recovery ranged from 96.89% to 99.73%. The lead determination results were all within the normal reference ranges specified by the three certified reference materials for bovine blood samples. Conclusion The matrix-matched calibration curve correction-ICP-MS is suitable for high-throughput determination of blood lead.

Aim To determine the effects and mechanisms of isoflurane on energy metabolism during ischemia and reperfusion in isolated rat hserts.Methods The rat Langendorff model was used,and isolated per-fused rat hearts were separated into untreated,isoflurane,chelerythrine(PKC inhibitor)plus isoflurane,and chelerythrine groups.All the hearts were subjected to treatment before ischemia,followed by 30 min of ischemia and 60 min of reperfusion.Hemodynamic variables were recorded,and metabolites were measured by high-performance liquid chromatography,and analyzed subcellular localization of PKC isoforms by Western blot analysis.Results The recovery of left ventricular developed pressure after ischemia was(33?8)%,(56?9)%,and(30?5)% in the untreated,isoflurane,and isoflurane with chelerythrine groups respectively.Compared with the untreated hearts,isoflurane significantly improved the recovery of left ventricular developed pressure(P

Objective To investigate the role of phosphatidyl-inositol 3-kinase-Akt (PI3k-Akt) signal pathway in the attenuation of ischemia-reperfusion (I/R) injury by sevoflurane preconditioning in isolated rat hearts. Methods Ninety-six adult male SD rats weighing 220-280 g were randomly divided into 6 groups ( n = 16 each): sham operation group (group S); I/R group; sevoflurane preconditioning group (group SP); wortmannin group (group W); dimethyl sulfoxide (DMSO) group (group D) and sevoflurane preconditioning + wortmannin group (group SW) . Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5%C02 at 37 ℃ . The hearts were continuously perfused for 180 min in group S. After 15 min of equilibration, the isolated hearts were subjected to 30 min of ischemia followed by 120 min of reperfusion in SP, W, D and SW groups. Croups SP, W, D and SW received 10 min of perfusion with K-H solution containing 2. 4% sevoflurane, 100 nmol/L wortmannin, 20 μmol/L DMSO, and 2.4% sevoflurane + 100 nmol/L wortmannin, respectively, followed by 5 min washout before I/R. Eight hearts in each group were selected and HR, left ventricular end-diabetic pressure (LVEDP), left ventricular developed pressure (LVDP), and ± dp/dtmax were recorded at the end of equilibration and at 15 min of reperfusion, Myocardial tissues were obtained at 15 min of reperfusion for determination of apoptosis (by TUNEL) and phosphorylated Akt (p-Akt) expression (by Western blot) . Another 8 hearts were selected at 120 min of reperfusion for determination of myocardial infarct size by TTC staining. Result Compared with group S, LVDP and ± dp/dt,^ were significantly decreased and LVEDP was significantly increased in groups I/R, SP, W, D and SW, and myocardial p-Akt expression was up-regulated in groups I/R, SP and D ( P ＜ 0.05). Compared with group I/R, LVDP and ± dp/dtmax were significantly increased, LVEDP and apoptosis index were significantly decreased, myocardial p-Akt expression was up-regulated, and myocardial infarct size was significantly reduced in group SP (P ＜0.05) . Conclusion Activation of PI3K-Akt signal pathway is involved in the attenuation of I/R injury by sevoflurane reconditioning in isolated rat hearts.

The central corneal thickness (CCT) in age 48 years or less of Chinese was characterized and its relationship with gender, age, refraction and intraocular pressure (IOP) was investigated. Right eyes of 1669 participants were included in this study (880 men, 52.7% and 789 women, 47.3%). Mean age of the samples was 23.8 +/- 5.9 years. After the examination of corneal topography and refraction, Goldman applanation tonometry was carried out by one physician. Tonometric values were the mean of three consecutive readings. Subsequently, another physician carried out ultrasonic pachymetry with the DGH 2000 AP ultrasonic pachymeter. Six measurements were made at the center of the cornea of each eye. The mean value was used for analysis. The results showed that mean CCT of male participants was 551.33 +/- 34.62 microm, 5.79 microm more than that of female participants. Linear regression analyses revealed that CCT was negatively related with age only in female and no association was found between refractive status and CCT. IOP was positively related to CCT, and there was a difference in IOP of 1.5 mmHg (1 mmHg = 0.133 kPa) per 100 microm difference in CCT. Ocular hypertension group was prone to have thicker cornea than average. The results indicated that in adult Chinese CCT tended to decrease with aging in female only. IOP measured by Goldmann tonometry was positively related with CCT so that CCT should be measured along with IOP.
Age Factors ; China ; Cornea/*anatomy & histology ; Corneal Topography ; Intraocular Pressure ; Reference Values ; Sex Factors ; Tonometry, Ocular

Objective : To establish a microwave-assisted digestion-inductively coupled plasma mass spectrometry (ICP-MS) with an octopole reaction system for simultaneous determination of six heavy metals in peanuts, including Cr, Ni, As, Cd, Pb, Hg. Methods : Peanut samples were shelled and crushed evenly, and 0.350 0 g was accurately weighed and digested with 5 mL nitric acid and 1 mL hydrogen peroxide in a digestion tank. Following microwave-assisted digestion, pure water was used to quantify the samples, and internal standards and an octopole reaction system were used to remove the interference. Then, the contents of six heavy metals were determined in peanuts by ICP-MS. The accuracy and precision of ICP-MS were evaluated using national criteria ( GBW 10013 and GBW 10044 ) and spike-and-recovery testing. Results : The six heavy metals showed good linearity at the selected linear range ( r≥0.999 8 ). The detection limits of ICP-MS ranged from 0.001 4 to 0.023 8 ng/mL, and the spike-and-recovery rates ranged from 94.7% to 98.8%, with the relative standard deviations ranging from 0.7% to 3.6%. In addition, the determination results of the standard reference materials were all within the normal reference range. The detection of six heavy metals was 100.0% in 60 peanut samples, and the contents of six heavy metals were all low. Conclusion The established ICP-MS assay is feasible for simultaneous determination of multiple heavy metals in peanuts.

AIM:?To?investigate?if?alfentanil?protects?the?isolated?rat?heart?against?myocardial?reperfusion?injury?and?if?the?mechanism?of?this?protection?is?mediated?via?opioid?receptors?and?NO-dependent?pathways. METHODS: Langendorff rat hearts were perfused at constant pressure with Kreb-Henseleit(K-H) buffer for 20 min?and?then?were?perfused?with?test?solution:?K-H?buffer?or?K-H?buffer?containing?alfentanil? 50 ?g?L -1,?alfentanil? 100 ?g?L -1, naloxone 200 ?g?L -1, alfentanil 100 ?g?L -1+naloxone 200 ?g?L -1, L-NAME 100 ?mol?L -1 and alfentanil 100 ?g?L -1+L-NAME 100 ?mol?L -1. After 10 min of this, the hearts were subjected to 25 min normothermic( 37 ℃) global ischemia followed by 30 min reperfusion with the same test solution as before. To evaluate myocardial function, LVEDP, LVDP, ?dp/dt max, HR and CF were measured at the 20th, 25 and 30th minute of perfusion and the 1st, 3rd, 5th, 10th, 20th and 30th minute of reperfusion. After experiment, the NOS and ATP content of myocardium were assessed. RESULTS: Before ischemia, alfentanil 100 ?g?L -1 decreased the HR at the 30th minute compared with the 20th minute(P