AIM: To study the protective effects of shenfu injection on myocardial ischemia/reperfusion (I/R) injury in rats and its potential mechanisms. METHODS: Myocardial ischemia/reperfusion was produced by tying and untying of left anterior descending coronary artery. Ischemia lasted for 30 min and reperfusion for 60 min. Twenty-four healthy male SD rats weighing 230-280 g were randomly divided into three groups (n=8 in each): sham-operation group, I/R group, and shenfu group which the shenfu injection (10 ml?kg -1) were injected intraperitoneally 30 min before ischemia. The plasma concentration of tumor necrosis factor-? (TNF-?), interleukin-6(IL-6) were measured by ELISA. The heart was harvested and levels of the nuclear factor kappaB (NF-?B) activity were determined by Ecl-western blot analysis and ultrastructures were observed by electron microscopy. RESULTS: NF-?B binding activity in myocardial nuclear and the plasma concentration of IL-6, TNF-? were significantly increased in I/R group than that in the sham-operation group (P

Objective To investigate the protective effects of Shenfu injection on myocardium against ischemia and reperfusion injury in rats. Methods Twenty-four healthy male SD rats weighing 230-280 g were randomly divided into three groups (n = 8,each): sham-operation group(Sham), ischemia and reperfusion group (I/R), Shenfu injection group(SF) . In Sham group, the anterior descending branch of left coronary artery was exposed and a piece of silk thread was placed around the artery but untied. The ischemia and reperfusion injury models in I/R and SF groups were made by temporary ligation of the anterior descending branch of left coronary artery,ischemia lasted for 30 min and reperfusion for 60 min. In SF group, the Shenfu injection(10 ml/kg) was intraperitoneally injected 30 min before ischemia. In Sham and I/R groups, normal saline (10 ml/kg) was intraperitoneally injected. After 60 min reperfusion, the blood samples of all rats in each group were collected from the left carotid arterial catheter for determination of the concentrations of plasma TNF-?, IL-6 ( ELISA) . The myocardium samples were obtained for ultrastructure observation (Electron-microscope) .Results Compared with that in Sham group, the plasma concentrations of TNF-? and IL-6 in I/R group significantly increased( P

Objective To investigate the effect of Shenfu injection (SFI) on hypoxia and reoxygenation (H/R)-induced apoptosis and the expression of Bcl-2 and Caspase-3 in cultured neonatal rat cardiomyocytes and to explore the possible molecular protective mechanisms of SFI from hypoxia and reoxygenation injury in cardiacmy-ocytes in vitro. Method The experiment was performed in Research Center for Cardiovascular Regenerative Medicine, Cardiovascular Institute and Fuwai Hospital in Beijing. Ventricular myocytes from the hearts of neonatal Sprague-Dawley rats (1- to 2-day old) were cultured. The model of hypoxia and reoxygenation injury was devel-oped in primary cultured neonatal rat cardiacmyocytes. The cultured cells were randomly divided into four groups: (1) Control group (Con group), without any treatment; (2) Hypoxia and Reoxygenation group (H/R group),4 h hypoxia followed by 16 h reoxygenation; (3) Low-dose SFI group (L-SFI group),cardiacmyocytes were pretreated with a low dose (50 μL/mL) of SFI for 30 min followed by H/R; (4) High-dose SFI group (H-SFI group),car-diacmyocytes were pretreated with a high dose (100 μL/mL) of SFI for 30 min followed by H/R. Apoptosis was quantified by fluoreacence-activated cell sorter (FACS) analysis after staining with Fluorescein isothiocyanate (FITC)-labled Annexin-V (Annexin V-FITC) and propidine iodide (PI). The expressions of Bcl-2 and Caspase-3 were detected by ECL-Western blot analysis. All data are expressed as mean±S.E.M. One-way analysis of vari-ance (ANOVA) was performed followed by Student-Newman-Keul test using SSPS 11.5 software. A p value less than 0.05 were considered as statistically significant. Results The results of FACS analysis indicated that the rate of different apoptotic process in cardiomyocytes was significantly increased after H/R, while after SFI treatment the occurrence of cell apoptosis induced by H/R was decreased significantly. The results of ECL-Western blot analysis showed that cells' exposure to H/R induced proteolytic cleavage of caspases,as revealed by the appearance of the characteristic fragment at 17 000 of Caspase-3 and this proteolytic activation was nearly completed with difference concentration SFI incubation. The anti-apoptotic protein Bcl-2 in cardiomyocytes was decreased after H/R insult and was increased in cells with SFI pretreatment. Conclusions SFI has protective effects on cardiacmyocytes a-gainst apoptosis that could be induced by H/R injury, the mechanisms of which probably involve the inhibition of down-regulation of Bcl-2 protein level and sequential activation of Caspase-3.

Objective To evaluate the effects of heme oxygenase-1 (HO-1) mediated by cell penetrating peptide PEP-1 on liverinjury induced by intestinal ischemia/reperfusion (I/R) in rats.Methods Twenty-four male Sprague-Dawley rats,aged 7-9 weeks,weighing 210-260 g,were randomly divided into 3 groups (n =8 each):sham operation group (group S),intestinal I/R group (group I/R) and PEP-1/HO-1 group (group HO).To establish a model of intestinal I/R,intestines were exteriorized and the superior mesenteric artery was exposed and occluded for 45 min ischemia,and then the clamp was removed for 120 min reperfusion.The PEP-1/HO-1 fusion protein 0.5 mg was injectedvia ihe left iliac vein 30 min prior to ischemia in group HO.The superior mesenteric artery was only exposed but not occluded in group S.At the end of reperfusion,blood samples were collected from the right common carotid artery for measurement of serum aspartate aminotransferase (AST),alanine aminotransferase (ALT) activities.The rats were then sacrificed and livers were removed for microscopic examination and for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in livertissues.Results Compared with group S,serum AST and ALT activities and MDA content in liver tissues were significantly increased,while SOD activity in liver tissues was decreased in groups I/R and HO (P ＜ 0.05).Compared with group I/R,serum AST and ALT activities and MDA content in liver tissues were significantly decreased,while SOD activity in liver tissues was increased in group HO (P ＜0.05).Liver injury induced by intestinal I/R was significantly attenuated in group HO compared with group I/R (P ＜ 0.05).Conciusioon HO-1 protein mediated by cell penetrating peptide PEP-1 can attenuate liver injury induced by intestinalI/R in rats.

Objective To assess the behavior of children with tic disorder (TD),and to analyze the behavioral characteristics among children with TD at different clinic conditions.Methods Sixty-three children with TD were evaluated with Child behavior checklist (CBCL).ANOVA and t-test were used to analyze the difference in the total and individual scores of CBCL in the children classified according to the different clinical types,the severity of TD,and comorbid attention-deficit hyperactivity disorder(ADHD).Results There were no significant differences among the total and individual scores of CBCL in the patients of the different clinic types( P ＜ 0.05 ) ;the scores of body complain in the patients in moderate to severe conditions (4.15 ± 2.34) were higher than that of those in mild condition ( 2.68 ± 2.22 ) ( t =- 2.540,P =0.014) ; the scores of attention problem (9.94 ± 3.57 ),disciplinary offence ( 3.94 ± 3.06 ),aggressive behavior ( 15.39 ± 5.12 ),exportoriented behavior problems ( 13.98 ± 7.34)and behavior problem (47.89 ± 17.51 )in TD comorbid ADHD were higher than in simple TD group ( 7.31 ± 3.34,2.44 ± 2.22,7.24 ± 4.93,9.78 ± 6.55,37.07 ± 17.98 ) ( t =- 2.774,- 2.166,- 1.930,- 1.956,- 2.174,P =0.007,0.034,0.048,0.04 1,0.034 ).Conclusion Children with TD at different clinical conditions have varied behavioral problems and behavioral characteristics,while comorbid ADHD is the most significant factor to affect TD patient's behaviors.

Objective To investigate the effects of penehyclidine hydrochloride ( PHC) on lipopolysaccharide (LPS) -induced endothelial cells injury and its mechanism. Methods ECV-304 was cultured in RPMI1640 in a 5% humidified CO2 atmosphere at 37 ℃. Then cultured cells were used to assess the following treatments: control group, LPS group (1 μg/mL) and PHC group(2 μg/mL). At the end of the experiments, supernatant was collected for determination of lactate dehydrogenase ( LDH), and cells were collected for determination of malondialdehyde (MDA), superoxide dismutase (SOD), and nitric oxide (NO) levels. And extracellular regulated kinasel/2( ERKl/2)and JNK MAPK (mitogen-activated protein kinases, MAPK) protein expressions were determined using Western blot technique. Analysis of variance (ANOVA) was used for statistical analysis to compare values among all groups. A significant difference was presumed for a probability value < 0.05. Results Compared with control group, LDH leakage [(1642 ± 367) U/L vs (169±33)U/L], the contents of MDA[(13. 2 ± 1. 2) nmol/L vs (7. 2 ±0. 8)nmol/mL] and NO levels [(143.2 ± 10.3) μmol/L vs(85.5 ±4.1) μmol/L], expressions of ERK1/2 and JNK were remarkably increased and SOD activities[(41.2 ±2.7) U/mL vs (61. 1 ±2.8) U/mL] were obviously decreased in LPS group. PHC markedly decreased LDH leakage [(392 ±90) U/L], MDA contents [(8. 6 ± 1. 3) nmol/ mL] and NO levels [(92.1 ±6.6) μmol/L], ERK1/2 activation and enhanced SOD activities [(58.0± 3.0) U/mL]. Conclusions PHC could protect endothelial cells against LPS-induced cell injury. The effect of PHC is likely mediated through inhibition of ERK1/2 MAPK activation.

Objective To investigate the effect of penehyclidine hydrochloride (PHC) on iNOS expression in lung in septic mice.Methods Female KM mice were subjected to cecal ligation and puncture (CLP,a model of polymicrobial sepsis) or shame operation.Thirty-six KM mice weighing 20～25 g were randomly assigned into three groups:shame CLP group,CLP group and PHC group.Mice in PHC group were given PHC 0.45 mg/kg intraperitoneally at 1 h before CLP. Six hours after CLP,the lungs of each group were sampled for light and electronic microscopy.The expression of iNOS mRNA in lung tissue were detected by in situ hybridization.The survival rates were observed at 24 h after the operation. Results The CLP group was observed thickened alveolar septa,as well as mitochondrial cristae swelling and mitochondrial vacuolation under electronic microscope.Emptied lamellar bodies could be also found.Histology of lung in PHC group had little changed.Expression of iNOS in lung in PHC group was significantly lower than that of the CLP group.At 24 h after CLP challenge,70.0% of the PHC mice lived,remarkably increased compared with that of the CLP group (26.7%),P＜0.05.Conclusion PHC had effective effect for increasing the survival rates of septic mice,inhibiting the expression of iNOS and reducing the severity of lung injury.

Objective To investigate the humoral and cellular immune responses in patients with acute brucellosis, and evaluate dynamic changes of regulatory T-lymphocytes (Foxp3+ Treg) in the peripheral blood of patients during treatment, in order to clarify the relationship between immunosuppression and the therapeutic effect in human brucellosis.Methods Sixty-five patients with brucellosis hospitalized at the Third Department of Infectious Diseases, Heilongjiang Agriculture and Reclamation Bureau General Hospital between July 2015 and November 2015 were included.Twenty-eight patients were treated with conventional therapy (group A: patients received 3 courses of treatment.Each lasted for 20 days with one-week interval), and 37 patients were treated with conventional therapy in combination with immunopotentiator (group B).Thirty healthy volunteers were enrolled as the controlled group.The ratio of CD3+CD4+ Foxp3+ Treg cells in the peripheral blood of brucellosis patients were measured by flow cytometry (FCM) at the end of each course of treatment.Data in accordance with normal distribution were described as mean±standard deviation.Comparison between two groups was done by two sample t test.Comparison among multiple groups was performed by analysis of variance and SNK test.Data that did not fit the normal distribution were analyzed by multiple-sample nonparametric test.Results After the first (20 d), second (50 d) and third course of treatment (80 d), the ratios of Foxp3+Treg in the peripheral blood of 65 acute brucellosis patients were 2.83%, 3.77% and 4.03%, respectively, which were all significantly higher than control group (1.69%;t=5.97, 9.05 and 5.66, respectively, all P<0.01).At the end of the first course of treatment, the ratios of Foxp3+Treg in group A and B showed no statistically difference (t=0.33, P>0.05), while those were both higher than control group (t=7.09 and 4.94, respectively;both P<0.01).At the end of the second course, the ratio of Foxp3+ Treg in group B was higher than group A (t=2.22, P<0.01), and both of them were higher than control group (t=10.79 and 7.25, respectively;both P<0.01).At the end of treatment, Foxp3+ Treg in group A was also significantly higher than the other two groups (t=6.02 and 6.45, respectively;both P<0.01).Conclusions In patients with acute brucellosis treated with the standard antibiosis treatment in combination with immunopotentiator, the ratio of Foxp3+Tregs significantly increases and maintains at a high level, which suggests that extra immunopotentiator may be not helpful for the treatment of brucellosis at the very early stage.

Objective To evaluate the effects of dexmedetomidine or dezocine alone or the combination of the two agents on the emergence agitation in patients undergoing thoracic surgery.Methods Eighty ASA Ⅰ or Ⅱ patients,aged 18-64 yr,weighing 48-75 kg,scheduled for elective thoracic surgery under general anesthesia,were randomly divided into 4 groups (n =20 each):control group (group C),dexmedetomidine group (group DEX),dezocine group (group DEZ) and dexmedetomidine + dezocine group (group DEX + DEZ).In group DEX,dexmedetomidine 0.5 μg/kg was injected intravenously at 15 min before induction of anesthesia,followed by continuous infusion of dexmedetomidine at 0.4 μg· kg-1 · h-1 until beginning of chest closure.Dezocine 0.1 mg/kg was injected intravenously after beginning of chest closure in group DEZ.In group DEX + DEZ,dexmedetomidine 0.5 μg/kg was injected intravenously at 15 min before induction of anesthesia,followed by continuous infusion of dexmedetomidine at 0.4 μg·kg-1 ·h-1 until beginning of chest closure and then dezocine 0.1 mg/kg was injected intravenously.While the equal volume of normal saline was given instead in group C.Flurbiprofen axetil 50 mg was injected intravenously at beginning of skin closure in each group.Venous blood samples were collected for determination of plasma C-reactive protein (CRP),TNF-α and IL-10 levels at 10 min before induction of anesthesia (T1),before skin closure (T2),immediately after extubation (T3) and 15 min after extubation (T4).Side effects such as agitation during emergence from anesthesia were recorded.Sedation was assessed using Ramsay score.Results Compared with group C,the levels of plasma CRP and TNF-α at T2-4 and ratio of TNF-α/IL-10 were significantly decreased,the levels of IL-10 were increased at T2-4,the degree and incidence of agitation were decreased,and Ramsay score was increased in the other three groups (P ＜ 0.05).Compared with groups DEX and DEZ,the levels of plasma CRP and TNF-α at T2_4 and ratio of TNF-α/IL-10 were significantly decreased,the levels of IL-10 were increased at T2-4,and the degree and incidence of agitation were decreased in group DEX + DEZ (P ＜0.05).No side effects such as hypotension,bradycardia,respiratory depression,nausea and vomiting were observed in the four groups.Conclusion Dexmedetomidine or dezocine alone or combination of the two agents can decrease the degree and occurrence of emergence agitation and inhibit the inflammatory response simultaneously,and the combination of the two agents provides better efficacy than either alone in patients undergoing thoracic surgery.

Objective To investigate the protective effects of heme oxygenase-1 (HO-1) mediated by cell penetrating peptide PEP-1 on myocardium against ischemia/reperfusion (IR) injury in isolated rat hearts. Methods Healthy male SD rats weighing 220-280 g were anesthetized with intraperitoneal pentobarbital. Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5% CO2 at 37 ℃. Eighteen isolated rat hearts were randomly divided into 3 groups ( n = 6 each): Ⅰ group sham operation (group S);Ⅱ group IR and Ⅲ group PEP-1/HO-1 + IR (group HO-1). The isolated rat hearts were perfused with an oxygena-ted (95% O2-5% CO2 ) K-H solution at 37 ℃ in a Langendorff apparatus and were subjected to 40 min of global ischemia followed by 50 min of reperfusion after 30 min of stabilization. In group Ⅲ (group HO- 1 ) the isolated hearts were perfused with 50 μmol/L PEP-1/HO-1 for 15 min before ischemia. After 50 min of reperfusion, HO-1expression, MDA content and SOD activity in myocardial tissues were determined. The activities of creatine kinase (CK) and lactic dehydrogenase (LDH) in coronary effluent fluid were measured. Results The HO- 1 expression was significanfly higher in HO-1 group than in group IR. IR induced significant increase in MDA content and decrease in SOD activity in myocardium and CK and LDH activities in coronary effluent in group Ⅱ compared with group S. PEP-1/HO-1 significantly attenuated IR-induced changes. Conciusion HO-1 mediated by PEP-1 has protective effects on myocardium ngainst IR injury in rats.