OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

Objective To study the effects of nitric oxide on Ca2+-activated K+ (KCa) channels in the hypothalamus neurons of newborn SD rats. Methods The data were recorded using inside-out or cell-attached configuration of patch-clamp technique and the kinetic changes of KCa channels are compared before and after 100 μmol/L sodium nitroprusside (SNP) was added into the bathing solution. Results When 100 μmol/L SNP was added to the bathing solution in cell-attached configuration, the open probability of the KCa channels increased from (7.3±1.5)% to (40.2±6.5)%, open time from (7.12±1.41) ms to (15.34±3.45) ms, frequency from (11.3±3.5) Hz to (26.6±4.2) Hz. Conclusion NO-cGMP signal pathway could greatly increase the open probability of the channels as a result of both the prolongation of open period and increase of open frequency, but the pathological or physiological roles it may play require further study.

Objective To study the effects of nitric oxide on Ca2+-activated K+ (KCa) channels in the hypothalamus neurons of newborn SD rats. Methods The data were recorded using inside-out or cell-attached configuration of patch-clamp technique and the kinetic changes of KCa channels are compared before and after 100 μmol/L sodium nitroprusside (SNP) was added into the bathing solution. Results When 100 μmol/L SNP was added to the bathing solution in cell-attached configuration, the open probability of the KCa channels increased from (7.3±1.5)% to (40.2±6.5)%, open time from (7.12±1.41) ms to (15.34±3.45) ms, frequency from (11.3±3.5) Hz to (26.6±4.2) Hz. Conclusion NO-cGMP signal pathway could greatly increase the open probability of the channels as a result of both the prolongation of open period and increase of open frequency, but the pathological or physiological roles it may play require further study.

Objective To explore the perinatal outcomes of women with pulmonary hypertension complicating congenital heart disease(CHD).Methods Clinical data of 45 cases of pregnant women with pulmonary hypertension complicating CHD from Apr 1995 to May 2007 were analyzed and they were divided into three groups:29 cases of slight group[pulmonary hypertension of 30 mm Hg(1 mm Hg=0.133 kPa) to 49 mm Hg],8 cases of moderate group(pulmonary hypertension of 50 mm Hg to 79 mm Hg)and 8 cases of severe group(pulmonary hypertension equal to or higher than 80 mm Hg).The types of CHD,cardiac functional status(New York heart association,NYHA),gestational weeks of pregnancy termination,mode of delivery,pregnancy after CHD operation and outcomes of infants were compared between the groups. Results(1)The highest incidence of CHD were atrial septal defect and ventricular septal defect(58%, 26/45).The rate of pregnant women after CHD operation was 29%(13/45),they were mainly in slight group and their NYHA class were in Ⅰ-Ⅱ.(2)The occurrence rate of NYHA class Ⅲ-Ⅳ was 7/8 in severe group.The rate of NYHA class Ⅰ-Ⅱ as 6/8 in moderate group.The rate of NYHA class Ⅰ- was 97%(28 /29)in slight group.(3)The rate of term delivery was 93%(27/29),preterm labor 3% (1/29),abortion 3%(1/29),and the birth weight was(3153?399)g on average in slight group.The rate of term delivery was 5/8,preterm labor occurred in 3 cases in moderate group.The rate of term delivery was 5/8,preterm labor occurred in 2 cases,and iatrogenic abortion in 1 case in severe group.The average birth weight between slight group and moderate or severe group had a significant difference.(4)Caesarean section rate was 78 %(35/45)among all patients.The rate of cesarean section delivery was 76%(22/29)in slight group,6/8 in moderate group,and 7/8 in severe group.(5)The rate of pregnant women who had portent heart failure or heart failure was 24%(11/45),overall maternal mortality was 4%(2/45).Conclusions The higher the pulmonary hypertension,the worse the outcome of the mother and fetus;The pregnant women with good heart function after cardiac operation would have a good perinatal outcome.Cesarean section is more suitable for those women.

Adolescent ; Arrhythmias, Cardiac ; etiology ; Cardiac Catheterization ; Child ; Child, Preschool ; Electrocardiography ; Female ; Heart Septal Defects, Atrial ; therapy ; Humans ; Infant ; Male

OBJECTIVETo evaluate the feasibility of angiography combined with transthoracic echocardiography (TEE) as a modified management of the transcatheter occlusion of patent ductus arteriosus (PDA).

METHODSForty children with PDA were randomly divided into two groups (n=20 each): observed and control. The control group accepted traditional transcatheter occlusion, and the observed group received a modified management (angiography combined with TEE). The children in the observed group were monitored by realtime TTE.

RESULTSA complete occlusion was acquired by one occlusion operation in each child in the observed group. The TTE demonstrated that the occlusion device was in place, and that the blood flow velocities in the left and right pulmonary artery and the descending aorta were in normal ranges. There were shorter X-ray exposure time, shorter recovering time and less ICU stay time in the observed group than in the control group. The complications associated with blood vessel puncturation occurred in four children from the control group, but none of the observed group had the complications. The total hospitalization cost in the observed group was less than in the control group.

CONCLUSIONSAngiography combined with TEE as a modified management of the transcatheter occlusion of PDA is recommended.

Adolescent ; Cardiac Catheterization ; Cardiac Surgical Procedures ; methods ; Child ; Child, Preschool ; Ductus Arteriosus ; diagnostic imaging ; Ductus Arteriosus, Patent ; diagnostic imaging ; surgery ; Echocardiography ; Humans ; Infant ; Radiography

Natural killer (NK) cells can suppress the development of graft vs host disease (GVHD) while retaining antitumor response in allogeneic hematopoietic stem cell transplantation (allo-HSCT). Sphingosine-1-phosphate receptor 5 (S1PR5) can regulate NK cell migration and distribution in vivo by interacting with sphingosine-1-phosphate (S1P). This study was aimed to investigate S1PR5 expression change of NK cells in allo-HSCT and to explore the relationship between S1PR5 change and frequency of acute/chronic graft-versus-host disease (aGVHD/cGVHD). The S1PR5 expression was detected by real time quantitative PCR in the RNA extracted from blood NK cells of 17 couples of donor and recipient one month after allo-HSCT. The results showed that S1PR5 mRNA level variations in NK cells of donors and recipients post-allo-HSCT were not statistically significant (0.235 ± 0.191 vs 0.330 ± 0.261, P > 0.05). S1PR5 expression of NK cells was significantly lower in patients with aGVHD than those in patient without aGVHD (0.973 ± 0.834 vs 6.166 ± 5.32, P < 0.05). Compared with the corresponding donor, S1PR5 expression levels of patient declined by more than 10 that caused the high incidence of aGVHD. No significant correlation was found between S1PR5 expression of NK cells and cGVHD (3.401 ± 2.324 vs 2.762 ± 1.972, P > 0.05). It is concluded that the decreased expression level of NK cells S1PR5 is associated with aGVHD occurrence. Possible mechanism is due to S1PR5 low expression affecting distribution of NK cells in vivo, so affecting the regulation of NK cells for aGVHD.
Graft vs Host Disease ; blood ; epidemiology ; metabolism ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Killer Cells, Natural ; metabolism ; Receptors, Lysosphingolipid ; metabolism ; Transplantation, Homologous ; adverse effects

The present study was aimed to explore the changes of phosphorylated AMP-activated protein kinase (pAMPK) level in skeletal muscle after exposure to acute hypobaric hypoxia and exhaustive exercise. Thirty-two male Sprague-Dawley (SD) rats were randomly divided into sea level and high altitude groups. The rats in high altitude group were submitted to simulated 5 000 m of high altitude in a hypobaric chamber for 24 h, and sea level group was maintained at normal conditions. All the rats were subjected to exhaustive swimming exercise. The exhaustion time was recorded. Before and after the exercise, blood lactate and glycogen content in skeletal muscle were determined; AMPK and pAMPK levels in skeletal muscle were detected by Western blot. The results showed that the exhaustion time was significantly decreased after exposure to high altitude. At the moment of exhaustion, high altitude group had lower blood lactate concentration and higher surplus glycogen content in gastrocnemius compared with sea level group. Exhaustive exercise significantly increased the pAMPK/AMPK ratio in rat skeletal muscles from both sea level and high altitude groups. However, high altitude group showed lower pAMPK/AMPK ratio after exhaustion compared to sea level group. These results suggest that, after exposure to acute hypobaric hypoxia, the decrement in exercise capacity may not be due to running out of glycogen, accumulation of lactate or disturbance in energy status in skeletal muscle.
AMP-Activated Protein Kinases ; metabolism ; Altitude ; Animals ; Computer Simulation ; Glycogen ; metabolism ; Hypoxia ; physiopathology ; Lactic Acid ; blood ; Male ; Motor Activity ; physiology ; Muscle, Skeletal ; enzymology ; metabolism ; Phosphorylation ; Physical Exertion ; physiology ; Rats ; Rats, Sprague-Dawley

Background There have been an abundance of literature on the analysis of the mechanical characteristic of the sclera at the entire seleral level in high myopia.However,some recent studies on high myopia are focused on the mechanical changes of the sclera on a cellular level. Objective This experiment was purposed to study how transforming growth factor-β2(TGF-β2)affected sclerotic desmocytes and the mechanical behaviors of scleral fibroblasts in the posterior part of the eyes in guinea pigs with experimental myopia. Methods Induced myopic animal models were established by wearing-10.00 D concave lens for 30 days in lateral eyes of 2-week-old guinea pigs.The fellow eyes were used as control group.Another 5 matched animals served as normal controls.The scleral fibroblasts of each group were purified with the tissue explant method and passaged for 2 generations in vitro.Cultured cells were identified by immunochemistry with vimentin,desmin,keratin and S-100 antibodies.Different concentrations of TGF-β2(0,1,10,100mg/L)were added into serum-free DMEM for 24 hours,and the viscoelastic properties of scleral fibroblasts were measured by micropipette aspiration technique. Results Compared with the fellow,eyes and normal control eyes,the equilibrium modulus and apparent viscosity in model eyes were significantly higher(P＜0.05).After treatment of TGF-β2,the equilibrium modulus and apparent viscosity in the model group and fellow eyes were positively correlated with the concentrations of TGF-β2(r=0.743,r=0.533,r=0.654,r=0.576,P＜0.05).Following the addition of 1 mg/L TGF-β2 and 10 mg/L TGF-β2,the equilibrium modulus and apparent viscosity of scleral fibroblasts were significantly reduced in model eyes compared with fellow eyes(P＜0.05).No significant difference was found in the equilibrium modulus and apparent viscosity of scleral fibroblasts between model eyes and fellow eyes after treatment with 100 mg/L TGF-β2(P＞0.05). Conclusion TGF-β2 car increase the mechanic indexes in a concentration.dependent manner.1 mg/L,10 mg/L TGF-β2 can lower the equilibrium modulus and apparent viscosity of scleral fibroblasts of normal eye and thus cause more changes in the mechanical behavior of scleral fibroblasts.

OBJECTIVETo verify the electric synchronism, mechanic synchronism and hemodynamics of selective site pacing.

METHODSPacing in the right ventricular cardiac apex (RVA), the right ventricular His bundle region (His), and the septum of right ventricular high-positioned outflow tract (RVOT), CO and CI were recorded. The electrical synchronism was assessed by observing the width and shape in a 12-lead surface ECG. The mechanical synchronism was estimated by using the VVI (vector velocity imaging) technology of the Acuson Sequia 512.

RESULTSThe results showed that CO and CI were lower while pacing in RVA, but they were not significant different (P>0.05). The QRS width: (124 +/- 5.3) ms while pacing in His, (144 +/- 7.1) ms while pacing in RVOT and (156 +/- 8.6) ms while pacing in RVA. The QRS width while pacing in His and in RVOT were narrower than in RVA and there were significant differences (P<0.01). Vector velocity imaging showed that mechanical synchronism was better while pacing in RVOT than that in RVA.

CONCLUSIONPacing in RVOT seems better than pacing in traditional RVA, and the operation was no more difficult than the traditional operation.

Adult ; Aged ; Bundle of His ; physiopathology ; Cardiac Pacing, Artificial ; methods ; Electrocardiography ; Female ; Heart Ventricles ; physiopathology ; Humans ; Male ; Middle Aged ; Pacemaker, Artificial