AIM:To study the effect of neuregulin-1 ( NRG-1β) or captopril alone and their combination on myocardial cell apoptosis and related gene expression in rats with chronic heart failure .METHODS:Young male Sprague-Dawley rats were randomly divided into sham-operated group, heart failure group, NRG-1βgroup, captopril group and combined treatment group .The volume overloaded heart failure model in the rats was established by aortocaval fistula .The cardiac function was evaluated by determining the changes of hemodynamics and plasma BNP level .The apoptotic index ( AI) of myocardial cells was assayed by TUNEL .The protein levels of p-Akt, Bax and Bcl-2 in left ventricular tissue were detected by Western blot .RESULTS: The LVEDP, AI and the expression of Bax were significantly lower in NRG-1βgroup, captopril group and combined treatment group than those in heart failure group ( P<0.05 ) , while the values of ±dp/dtmax and the protein levels of p-Akt and Bcl-2 were significantly higher in NRG-1βgroup, captopril group and com-bined treatment group than those in heart failure group (P<0.05).The effect of combined treatment group was more signifi-cant ( P<0.05) than that in NRG-1βgroup and captopril group .CONCLUSION: Neuregulin-1βor captopril alone and the combination of them effectively improve the cardiac function and inhibit myocardial cell apoptosis in chronic heart failure rats.The therapeutic effect of combination of NRG-1βand captopril is better than that of NRG-1βor captopril alone .

Objective To study the relationship between the polymorphism of interleukin-12B (IL-12B)gene and coronary heart disease.Methods We recruited 256 patients with coronary heart disease admitted to our department as the study group and 256 normal subjects as the control group.The polymorphism of IL-12B gene was detected by polymerase chain reaction and single nucleotide polymorphism.Coronary artery stenosis,visfatin,high sensitive C reactive protein and cardiac function were determined.Results The difference in rs15677380 and rs14050311 allele frequencies between the study group and the control group was significant (χ2 =6.19,7.24,P=0.045,0.021).The G allele of rs15677380 and C allele of rs14050311 were risk factors for coronary heart disease (OR=1.32,1.49).Conclusion IL-12B gene is associated with the occurrence and development of carotid atherosclerosis and participates in the development of coronary heart disease.

Objective To observe the influence of Simvastatin on immature rabbit model of chronic heart failure(CHF),and to explore the possible protective mechanism of Simvastatin for the rabbits with CHF.Methods Thirty immature male rabbits were divided into 3 groups randomly:control group,heart failure group (HF group)and Simvastatin group(SIM group),10 rabbits in each group.The models of CHF were established by injecting Adrinmycin via the auricular vein of rabbits (1.5 mg/kg,once 1 week,for 12 weeks).The control group were injected the same amount of 9 g/L saline.SIM group were given both injection of Adrinmycin and Simvastatin [1.5 mg/(kg · d)for 12 weeks].The immature rabbit's cardiac function and myocardial morphology changes were evaluated.The expressions of chromogranin A (CgA) and apoptosis signal-regulating kinase 1 (ASK1) were evaluated.Results (1) In control group,the immature rabbits were all alive;in the other 2 groups,most immature rabbits were depressed and sluggish.The survival rate of HF group was 60％,while in SIM group the survival rate was 80％.(2)Compared with the control group,the left ventricular ejection fraction in HF group and SIM group decreased significantly [(40.05 ± 6.74)％,(50.18 ± 5.73) ％ vs.(65.93 ± 5.65) ％,all P ＜ 0.01],while in SIM group was higher than that of HF group,and the difference was significant (P ＜ 0.05);compared with HF group,the left ventricular end diastolic diameter and left ventricular end systolic diameter decreased in SIM group [(13.48 ± 1.24) mm vs.(16.23 ± 2.82) mm;(9.87 ± 0.85) mm vs.(11.13 ± 1.21) mm],and the differences were significant (all P ＜ 0.05).(3) Compared with the control group,the expression of CgA (mean optical density 142.24 ± 17.14,127.93 ± 12.12 vs.78.65 ± 6.78,P ＜ 0.05;integrated optical density 1 422.41 ± 167.34,1 279.37 ± 118.15 vs.786.54 ± 75.84,P ＜ 0.05) and ASK1 (mean optical density 140.32 ± 18.65,115.48 ± 12.30 vs.69.85 ± 6.54,P ＜ 0.05;integrated optical density 1 403.23 ± 165.67,1 158.79 ± 137.81 vs.698.58 ± 64.51,P ＜ 0.05) increased in the HF group and SIM group.While the expression in the SIM group decreased significantly compared with that of the HF group,and the difference was significant (P ＜ 0.05).Conclusions Simvastatin can improve cardiac function of immature rabbits with CHF.The mechanism of SIM may depress the expressions of CgA and ASK1.

Objective To observe the effects of Simvastatin on myocardial apoptosis and oxidative stress mechanism in immature rabbits with chronic heart failure.Methods Thirty-six male New Zealand big-eared immature rabbits were randomly divided into 3 groups:Adriamycin(ADR) ± Simvastatin group(ADR-s group,n =12),in which ADR(1.5 mg/kg) received injection via the auricular vein of rabbits weekly,and the rabbits received oral Simvastatin [1.5 mg/(kg · d)] simultaneously for 12 weeks;ADR group (n =12),in which the rabbits received ADR like ADR-S group,and 9 g/L saline instead of Simvastatin;control group (CON group,n =12),which received the same amount of 9 g/L saline.Echocardiography examination was performed in 13th week.Myocardial fibrosis degree was detected by using MASSON staining,and the myocardial apoptosis was detected by using terminal deoxynucleotidyl transferase dUTP nick end labeling.Colorimetric method was used to detect the myocardial concentration superoxide dismutase (SOD) and malondialdehyde (MDA).Enzyme-linked immunosorbent assay was used to detect serum B-type brain natriuretic peptide (BNP) level.Results (1) In CON group,the immature rabbits were all alive.Four rabbits died in ADR group,and the survival rate was 66.7％,while 2 rabbits died in ADR-s group,and the survival rate was 83.3％.(2)Compared with CON group,the left ventricular end diastolic diameter (LVEDd) and left ventricular end systolic diameter (LVESd) in ADR-s group and ADR group increased [(11.90 ±1.09) mm,(ll.34 ±0.92) mm vs.(10.73 ±0.48) mm;(9.80 ±0.88) mm,(8.47 ± 1.23) mm vs.(7.31 ±0.36) mm];left ventricular fractional shortening(LVFS) and left ventricular ejection fraction(LVEF) decreased [(17.65 ± 1.70)％,(22.58 ± 2.19)％ vs.(31.79 ± 2.58) ％;(41.35 ± 3.19) ％,(49.17 ± 3.53) ％ vs.(64.34 ± 3.97) ％],and all the differences were significant(all P ＜ 0.05);LVEDd and LVESd in ADR-s group were lower than those of ADR group,while LVEF and LVFS in ADR-s group were higher than those of ADR group,and the differences were significant(all P ＜ 0.05).(3)MASSON staining:compared with ADR group,there was less myocardial cell hyperplasia of fibrous tissue in ADR-s group.(4) Compared with CON group,the apoptosis index was higher in ADR and ADR-s group [(34.25 ±11.13) ％,(24.00 ±6.85)％ vs.(16.58 ± 5.34)％],but ADR-s group had less than ADR group,and the differences were significant (all P ＜ 0.05).(5) Compared with ADR group,SOD activity of ADR-s group was higher [(13.40 ± 2.68) kU/L vs.(10.66 ± 2.99) kU/L],but MDA content was lower [(5.67 ± 1.36) μmol/mg vs.(7.08 ±0.98) μmol/mg],and the differences were significant (all P ＜0.05).(6) Serum BNP level in ADR group and ADR-s group was higher than that of the CON group[(33.28 ±9.58) μg/L,(26.71 ±6.72) μg/L vs.(13.56 ±2.82) μg/L],while was higher in ADR group than that of ADR-s group,and the differences were significant (all P ＜ 0.05).Conclusions Simvastatin can protect cardiac function of immature rabbits with chronic heart failure.The possible mechanism may be the up-regulation of myocardial SOD activity,reduction of cell lipid peroxidation and inhibition of myocardial apoptosis.

OBJECTIVE:To study the protective effects of soluble ornithine cyclase activator cinaciguat(called CIN for short) on human umbilical vein endothelial cells (HUVECs) under the condition of high glucose. METHODS:HUVECs were treated with glucose 33.3 mmol/L to induce oxidative stress injury model. HUVECs were divided into normal group,high glucose group and high glucose+CIN 0.01,0.1,1 and 10 μmol/L groups. The cell viability was detected by MTT assay,and the survival rate of cells was callulated. Besides,HUVECs were divided into normal group,normal+CIN 1 μmol/L group,high glucose group,high glucose+CIN 1 μmol/L group. MDA content and SOD activity were determined by TBA method and xanthine oxidase method;mRNA expression of ICAM-1 and VCAM-1 were determined by RT-PCR. RESULTS:Compared with normal group,survival rate decreased in high glucose group(P<0.05);compared with high glucose group,that increased in high glucose+CIN 0.01,0.1,1 and 10 μmol/L groups,in concentration-dependent manner;there was no statistical significance in survival rate between high glu-cose+CIN 1 μmol/L group and high glucose+CIN 10 μmol/L group (P>0.05). Compared with normal group,MDA content and mRNA expression of ICAM-1 and VCAM-1 increased in high glucose group,while SOD activity decreased(P<0.05);above indi-cators had no significant change in normal+CIN 1 μmol/L group(P>0.05). Compared with high glucose group,MDA content and mRNA expression of ICAM-1 and VCAM-1 decreased in high glucose+CIN 1 μmol/L group,while SOD activity increased(P<0.05). CONCLUSIONS:CIN can protect high glucose-induced HUVECs injury in vitro. The mechanism may be associated with in-hibition of oxidative stress and inflammatory response.

AIM: To investigate the protein patterns of bone marrow mesenchymal stem cells(MSCs) induced by 5-azacytidine and Shuanglong Formula(Radix et Rhizoma Ginseng,Radix et Rhizoma Salviae Miltiorrhizae) and molecular mechanism of induced differentiation was discussed at the level of proteome. METHODS: MSCs extracted from Chinese miniswine were induced by 5-azacytidine and Shuanglong Formula.Proteins were separated by two-dimensional gel electrophoresis,and were detected by silver staining method.The image was analyzed by PDQuest software,and the intensities of spots in the gels were used to quantify the expressions of proteins.Proteins of interest were chosen for in-gel digestion and MALDI-TOF-MS analysis,and then further identified by peptide mass fingerprinting(PMF). RESULTS: 16 altered proteins were identified by PMF and their regulation might be caused by the addition of medicate sera containing the active components of Shuanglong Formula.The effects of medicated sera containing Shuanglong Formula on the induced-differentiation could be discussed from the regulated expression of functional proteins; CONCLUSION: Shuanglong Formula may promote the 5-azacytidine-induced differentiation of bone marrow MSCs in vitro,and our study indicate that proteomics can be used for the analysis of molecular mechanism for different cell process.

Objective:To evaluate the effects of intranasal administration of glial cell line-derived neurotrophic factor (GDNF) on postoperative cognitive dysfunction in aged rats.Methods:Forty healthy Sprague-Dawley rats of both sexes, aged 21-23 months, weighing 480-600 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group S), operation group (group O), intranasal administration of low-dose GDNF group (group G1) and intranasal administration of high-dose GDNF group (group G2). Rats underwent exploratory laparotomy under anesthesia with chloral hydrate in O, G1 and G2 groups, while the rats in group S only received sham operation.The rats in group G1 and group G2 were intranasally treated with GDNF 25 and 50 μg (in 25 μl of PBS), respectively, and PBS 25 μl was nasally administered in group S and group O every day for 3 consecutive days after operation or sham operation.Morris water maze test was performed on days 3-7 after surgery, and then the rats were sacrificed, and hippocampal tissues were removed for determination of the expression of GDNF, high mobility group box 1 (HMGB1), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), activated caspase-3 and Bax (by Western blot). Results:Compared with group S, the escape latency was significantly prolonged on days 5-7 after operation, the number of crossing the platform was reduced, time spent in the target quadrant was shortened, expression of GDNF was down-regulated, and expression of IL-1β, TNF-α, HMGB1, activated caspase-3 and Bax in hippocampi was up-regulated in group O, and the number of crossing the platform was reduced, time spent in the target quadrant was shortened, and expression of IL-1β and TNF-α was up-regulated in G1 and G2 groups ( P<0.05). Compared with group O, the escape latency was significantly shortened on days 5-7 after operation, the number of crossing the platform was increased, time spent in the target quadrant was prolonged, expression of GDNF was up-regulated, expression of TNF-α, HMGB1, activated caspase-3 and Bax in hippocampi was down-regulated in G1 and G2 groups, and IL-1β in hippocampi was down-regulated in group G1 ( P<0.05). Compared with group G1, the expression of TNF-α in hippocampi was down-regulated ( P<0.05), and no significant change was found in the other parameters mentioned above in group G2 ( P>0.05). Conclusions:Intranasal administration of GDNF can improve postoperative cognitive dysfunction, and the mechanism may be related to inhibiting neuroinflammatory responses and neuroapoptosis in aged rats.

The aim of this study was to determine if the potassium aspartate and magnesium (PAM) prevent reperfusion-induced ventricular arrhythmias (RIVA) in ischemia-reperfusion (IR) rabbit heart. Thirty rabbits were randomly divided into control, ischemia and PAM groups. Arterially-perfused rabbit left ventricular preparations were made, and transmural ECG as well as action potentials from both endocardium and epicardium were simultaneously recorded in the whole process of all experiments. In control group rabbit ventricular wedge preparations were continuously perfused with Tyrode's solution, and in ischemia group and PAM groups the perfusion of Tyrode's solution was stopped for 30 min. Then the ischemia group was reperfused with Tyrode's solution and the PAM group with Tyrode's solution containing 2.42 mg/L PAM, respectively. ECG, QT interval, transmural repolarization dispersion (TDR) and action potentials from epicardium and endocardium were simultaneously recorded, and the RIVA of the wedge preparation was observed. Compared with control group, TDR and incidence of RIVA were significantly increased in ischemia group (P<0.05). The incidence of RIVA in control, ischemia and PAM group was 0/10, 9/10 and 1/10, respectively. Compared with ischemia group, TDR and incidence of RIVA were significantly reduced in PAM group (P<0.05). Potassium aspartate and magnesium significantly reduce TDR and prevent ventricular arrhythmia in ischemic rabbit heart.
Arrhythmias, Cardiac/etiology ; Arrhythmias, Cardiac/*prevention & control ; Myocardial Ischemia/*complications ; Myocardial Ischemia/physiopathology ; Myocardial Reperfusion Injury/*complications ; Potassium Magnesium Aspartate/*therapeutic use ; Random Allocation

OBJECTIVETo establish the method for the quality control of Xuefuzhuyu granule.

METHODThe content of Semen Armeniacae Amarum PE, Hydroxysafflor yellow A, paeoniflorin and ferulic acid in Xuefuzhuyu granule were determined by HPLC under multi-wavelength of 210, 403, 230, 316 nm.

RESULTThe calibration curve of Semen Armeniacae Amarum PE. hydroxysafflor yellow A, paeoniflorin and ferulic acid were linear within 0.1154-0.9232 (r = 0.9998), 0.0219-0.1754 (r = 0.9994), 0.3952-3.1616 (r = 0.9995), and 0.0426-0.3408 microg (r = 0.9998), respectively. The average recoveries were 101.78% (RSD was 1.7%), 99.60% (RSD was 2.9%), 98.90% (RSD was 2.0%), and 100.31% (RSD was 1.8%), respectively.

CONCLUSIONThe method of quantification is accurate, rapid and reliable for the quality control of Xuefuzhuyu granule.

Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; Chalcone ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; Drugs, Chinese Herbal ; analysis ; chemistry ; Glucosides ; analysis ; Monoterpenes ; Quinones ; analysis ; Reproducibility of Results

Objective To investigate the effects of intrathecal dexmedetomidine administration on the development of morphine tolerance and spinal inflammatory responses.Methods Thirty-three male SD rats weighing 180～200 g were randomly divided into 3 groups (n=11):Saline group (group NS),Morphine group (group M) and Dexmedetomidine group (group Dex).Animals of group NS were intrathecally injected with 10 tμL of saline daily for seven days;Animals in group M were intrathecally injected with 15 μg of morphine daily for seven days;Animals in group Dex were intrathecally injected with a mixture of 15μg morphine and 1.5 μg dexmedetomidine daily for seven days.At 1,3,5 and 7 day of intrathecal injection,hot water tail-flick test were used to evaluate analgesic response to thermal stimuli.After the last episode of behavioral test,Western blot analysis was applied to determine the protein levels of Iba-1 (microglial marker),IL-1β,TNF-a and phospho-p38MAPK (p-p38) in the spinal cord.In addition,microglia in the spinal cord was immuno-stained with anti-Iba-1 antibody and the densities of microglia were calculated.Results In group M and Dex,the values of maximal possible effect (MPE) in tail-flick test decreased gradually along with repeated morphine administration (P＜0.05).Compared with group NS,the values of MPE in tail-flick test at 1,3,5 and 7 day of morphine tolerance were higher in group M (P＜0.05).Compared with group M,the values of MPE in tail-flick test at 3,5 and 7 day of morphine tolerance were higher in group Dex (P＜0.05).Compared with group NS,the spinal protein levels of Iba-1,IL-1L TNF-α and p-p38 as well as the density of Iba-1 positive cells in group M were increased (P＜0.05).However,Compared with group M,the of Iba-1,IL-1β,TNF-α and p-p38 as well as the density of Iba-1 positive cells were decreased(P＜0.05).Conclusion Intrathecal dexmedetomidine administration can attenuate morphine tolerance by inhibiting microglia-mediated inflammatory responses in the spinal cord.