Objective To investigate a simple and dependable approach to establish the hypoxic-ischemic encephalopathy model in neonatal rat. Methods Twenty-one neonatal rats of 7 days old were randomly divided into control group,hypoxic group,and hypxic-ischemic group.Every group was randomly divided into 3 hours,6 hours,1 day,3 days,7 days, 14 days,and 21 days group,according to the time of killing.Left common carotid artery of neonatal rats at age of 7 days in hypoxic-ischemic group were ligated.Then,the rats in hypoxic and hypoxic-ischemic group were put in a state of 8% oxygen for 2.5 hours. Brain tissues of rats in 3 groups were observed with HE staining under light microscope.Results In hypoxic-ischemic group,there was found mild brain damage after hypoxic-ischomic 3 hours,the brain lesion was most severe at 1 day,glial cell proliferation was found at 3 days,much neur were losed at 14,21 days,and colloid scar was formed in cortex,striatum and hippocampi.Conclusion The method that left common carotid ontery of neonatal rats were ligated and then put in 8% oxygen for 2.5 hours is simple, rapid and dependable, which can be applied widely.

Objective To investigate the characters of culture of neural stem cells(NSCs) from neonatal rats in different states in vitro.Methods Forty-two neonatal rats at age 7 days were divided randomly into 3 groups as control group,hypoxic group and hypoxic-ischemic group,each having 14 rats.Forteen rats of every group divided randomly into 7 small groups,each including 3 h,6 h,1 d,3 d,(7 d),14 d and 21 d,according to the time to put to death,each having 2 rats.After builting rat models of hypoxic-ischemic encephalopathy,NSCs from hippocampus in 3 groups were isolated,then cultured,passed,differentiated and differentiated with single cell clone and immunocytochemistry tecnique.Results NSCs in hippocampus from 3 groups were cultured in form of typical neuraospheres in suspension.The cells could be cloned,passed continuously and induced.There were differences among 3 groups when primary NSCs were cultured at 3 h and 6 h time points.But at 1 d,3 d,7 d,14 d and 21 d time points,clony neuraospheres from primary NSCs in hypoxic group were the most among 3 groups while clony neuraospheres from primary NSCs in hypoxic-ischemic group were the lest.Conclusions NSCs from hippocampus of neonatal rats in different states remain to be cultured,meanwhile,NSCs are decreased with increase of age,elongation of illness course and progress of state of an illness.

Objective To evaluate the effect of staged repair for tetralogy of Fallot (TOF) associated with pulmonary artery hy- poplasty.Methods From June 1996 to June 2006,37 patients with TOF were operated on.There were 26 males and 11 females. Their age was 5 months to 17 years(mean 3.6 years) and weight was 4.6～38.0 kg.All patients were diagnosed as TOF with pulmo- nary artery hypoplasty by cardiac catheterization.The mean pre-operative arterial saturation of the patients was (68.2?6.5) %,Mc- Coon ratio was 0.95?0.26 (0.81～1.17) and Nakata index was 82.7?21.6(71.6～97.5) mm~3/m~2.At the time of the first surgi- cal procedure,17 patients underwent central aortopulmonary shunt,13 patients received modified Blalock-Taussig shunt in the left side and 7 patients had modified Blalock-Taussig shunt in the right side.Results There were no easly operative deaths and no late deaths after the first stage repair.Pleural effusion after shunt occurred in 5 patients.The mean arterial saturation was significantly increased to (91.3?10.4) %,P

OBJECTIVETo study the protective effect of hyperoxia solution on acute lung injury caused by phosgene poisoning by observing the changes of PaO2 and malondialdehyde (MDA) contents, superoxide dismutase (SOD) activity in serum and Glutathione (GSH/GSSG) contents in lung tissues.

METHODSThe rabbits were divided into normal control group, hyperoxia solution (H0) and balance salt (BS) groups. Group HO and Group BS inhaled phosgene and the former was given intravenously hyperoxia solution (which was replaced by balance salt solution in Group BS). The content of MDA and the activity of SOD in serum were observed at different time points, the amount of GSH and GSSG in lung tissue were also measured.

RESULTS(1) The serum MDA contents increased and PaO2, SOD activity decreased significantly in Group HO and Group BS along with time increasing as compared with control group. The contents of GSH in lung tissue decreased in two groups compared with that in control group, however the contents of GSSG ascended instead. (2) At 3 and 8 h of the experiment, PaO2 of Group HO [(9.91 +/- 0.49), (9.15 +/- 0.46) mm Hg respectively] were significantly higher than those of Group BS [(9.03 +/- 0.76), (8.11 +/- 0.57) mm Hg respectively] (P < 0.01). The contents of MDA of Group HO (3.66 +/- 0.35), (5.31 +/- 0.15) micromol/L respectively] were lower than those of Group BS [(4.32 +/- 0.26), (7.4 +/- 0.33) micromol/L respectively] (P < 0.01). SOD activity in Group HO [(237.37 +/- 29.96), (208.10 +/- 18.80) NU/ml respectively] were higher than those of Group BS [(195.02 +/- 21.44), (144.87 +/- 21.26) NU/ml respectively] (P < 0.05 or P < 0.01). The content of GSSG lung tissue in Group HO (423.67 +/- 38.21) micromol/L were lower than those of Group BS (523.85 +/- 43.14) mol/L (P < 0.01). There were no significant differences in the content of GSH in lung tissues between Group HO and group BS.

CONCLUSIONHyperoxia solution can reduce acute lung injury of rabbits following phosgene poisoning.

Acute Lung Injury ; etiology ; metabolism ; pathology ; Animals ; Glutathione Peroxidase ; metabolism ; Hyperoxia ; Lung ; drug effects ; metabolism ; pathology ; Malondialdehyde ; analysis ; Oxygen ; administration & dosage ; pharmacology ; Phosgene ; poisoning ; Rabbits ; Superoxide Dismutase ; metabolism

OBJECTIVETo study growth characteristics of neural stem cells (NSCs) from subventricular zone (SVZ) of the different human fetal brain at different gestational age and to provide experimental and theoretical evidences for clinical application of NSCs for treatment of certain diseases.

METHODSNinety human embryos at gestational age 16 - 36 weeks were collected and were divided into six groups according to gestational age: 16 w, 20 w, 24 w, 28 w, 32 w and 36 w. Each group had 15 embryos and brain tissues were taken from each embryo's SVZ. All subjects had congenital heart disease or digestive tract abnormity diagnosed with B ultrasound at antepartum, but none had abnormal development of brain. Pregnant mother and her husband desire termination of pregnancy. The morphology, existing mode and the number of neural stem cells in subventricular zone were examined with immunohistochemical method. The NSCs in subventricular zone were cultured, passaged and differentiated with cell culture technique, then were identified with immunohistochemical method.

RESULTSNSCs in SVZ from the different human fetal brain existed in a scattered manner in the network formed by stellate cells, NSCs had round, ellipse and fusiform shape, especially in stellate shape. NSCs had larger and smaller size and distributed in dense or scattered forms, each having zero to two enations, most had one or two. NSCs had less cytoplasm. The nucli of the NSCs had a round shape with loose chromatin and 1 - 4 nucleoli. Most of NSCs existed in singular scattered form, some of them showed symmetrical or asymmetrical division, some of them showed synaptic connection with other NSCs. The number of NSCs in SVZ from groups with different fetal age decreased with increasing gestational age (chi(2) = 4644.602, P < 0.01). NSCs in SVZ from the different human fetal brain cultured with serum-free medium formed typical neurospheres in suspension. The cells could be passaged continuously, and could express nestin antigen. Serum-contained medium induced neural stem cells to differentiate and express specific antigens of neuron, astrocyte and oligodendrocyte.

CONCLUSIONSNSCs existed in SVZ of human embryos at different gestational age. There are differences in morphology, existing pattern and the number of NSCs in SVZ at different gestational age. NSCs in SVZ at different gestational age may be cultured in vitro.

Age Factors ; Astrocytes ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Cerebral Ventricles ; cytology ; Female ; Fetal Stem Cells ; Fetus ; cytology ; Gestational Age ; Humans ; Immunohistochemistry ; Intermediate Filament Proteins ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Nestin ; Neurons ; Oligodendroglia ; Pregnancy

OBJECTIVETo establish a neonatal rat model of hypoxic-ischemic encephalopathy and clarify the changing features of neural stem cells (NSCs) in episodes of hypoxic-ischemic encephalopathy (HIE) so as to provide experimental and theoretical evidences for treating HIE by applying NSCs at the appropraite time.

METHODSTotally 210 neonatal rats aged 7 d were divided randomly into three groups, normal control group, hypoxic group and hypoxic-ischemic group with 70 rats in each. According to the time of sacrefice, 70 rats of every group were further divided randomly into seven groups including third hour (3 h), the sixth hour (6 h), first day (1 d), third day (3 d), the seventh day (7 d), the fourteenth day (14 d) and the twenty-first day (21 d), with 10 rats in each subgroup. The left common carotid artery of the neonatal rats in hypoxic-ischemic group was ligated and those in the hypoxic group were subjected to inhalation of 8% oxygen for 2.5 h. NSCs from brain tissues of the rats of the three groups were determined with HE staining and immunohistochemical method under light microscope.

RESULTSMost of neonatal rats in hypoxic-ischemic group behaved turning to the left stably 1 h after normal concentration of oxygen was given. In hypoxic-ischemic group, slight brain injury occurred at 3 h, severe brain injury appeared at 1 d, glial cells proliferated at 3 d and 7 d, brain atrophy was found at 14 d and 21 d. NSCs existed in brain tissues of rats in all the three groups. NSCs in normal control and the hypoxic group mainly distributed in hippocampus, subventricular tissues, striatum and cortex. But NSCs in hippocampus located in layers of molecule, cone cell and inner granular cell. NSCs in hypoxic-ischemic group showed obvious regional distribution, less in the regions with pathological changes. At 3 h, 6 h and 14 d, there was no difference in the number of NSCs between hypoxi and hypoxic-ischemic group. At 1 d, 3 d and 7 d, there was a highly significant difference in the number of NSCs between hypoxic and hypoxic-ischemic group. At 21 d, there was a significant difference in the number of NSCs between hypoxic and hypoxic-ischemic group, meanwhile, there was a significant difference in the number of NSCs between control and hypoxic group. At 3 d, there was a very significant difference in the number of NSCs between control and hypoxic-ischemic group. At 7 d and 21 d points, there was a highly significant difference in the number of NSCs between control and hypoxic group.

CONCLUSIONThe neonatal rat model of HIE was successfully established. NSCs increased in earlier period and decreased in later period of HIE, ultimately, NSCs located in the injured regions died one after anotner. Hypoxia induces NSCs' proliferation.

Animals ; Animals, Newborn ; Atrophy ; Brain ; pathology ; Carotid Artery, Common ; surgery ; Disease Models, Animal ; Hypoxia-Ischemia, Brain ; pathology ; Immunohistochemistry ; Ligation ; Multipotent Stem Cells ; pathology ; Neuroglia ; pathology ; Neurons ; pathology ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation ; methods ; Time Factors

BACKGROUNDSkeletal muscle dysfunction is one of important systemic manifestations of chronic obstructive pulmonary disease (COPD) and is associated with mortality in patients with COPD, thus quantifying its strength is of great clinical interest and of particular value. Quadriceps maximal volitional contraction (MVC) is often used for the routine measurements of this muscle's strength; while twitch tension (TwQ) evoked by magnetic stimulation of the femoral nerve has been employed for measurement of quadriceps strength non-volitionally. We aimed to investigate the prevalence and severity of skeletal muscle dysfunction in COPD patients by measurement of quadriceps strength with volitional and non-volitional techniques, and to probe into some methodological issues.

METHODSWe recruited 71 COPD patients and 60 control subjects. Quadriceps strength was measured with both maximality of TwQ and MVC force. The reproducibility for TwQ and MVC was investigated using within-occasion variability from three repeated maneuvers.

RESULTSMaximal TwQ was achieved in 121 participants at a mean of 90% of the stimulator's maximum output. The mean maximality of TwQ was decrease by about 44% - 47% in COPD patients as compared with controls (P < 0.05), so was MVC. There was a significant correlation between quadriceps TwQ and MVC, and the mean ratio of TwQ/MVC was 0.29 in controls and 0.33 in patients. The coefficient of variation showed that TwQ yielded lower within-occasion variability than MVC in both groups.

CONCLUSIONSQuadriceps strength is commonly and substantially impaired in patients with COPD, in terms of MVC as well as TwQ. The magnetic stimulation of the femoral nerve presents a higher reproducibility and is a better technique for measurement of quadriceps strength for the general population, especially for those who are too unwell to perform a full MVC; while it may not be applied to subjects who are over-weighted.

Aged ; Female ; Femoral Nerve ; physiology ; Humans ; Male ; Middle Aged ; Muscle Strength ; physiology ; Pulmonary Disease, Chronic Obstructive ; physiopathology ; Quadriceps Muscle ; physiology

BACKGROUNDChronic obstructive pulmonary disease (COPD) is associated not only with airway inflammation characterized by mucin hypersecretion but also with systemic inflammation. Tumor necrosis factor alpha (TNF-alpha) is found to take part in systemic inflammation, and ErbB3 plays an important role in mucin hypersecretion of COPD. Since TNF-alpha converting enzyme (TACE) is involved in the activation of both TNF-alpha and ErbB3, we established rat models of COPD to investigate the expressions of TACE, TNF-alpha and ErbB3 and to explore the correlations among TACE, TNF-alpha and ErbB3 respectively.

METHODSThirty Wistar male rats were randomly divided into COPD group (group C, n = 10), saline solution parallel group (group P, n = 8), and normal control group (group N, n = 8). Group C was challenged with passive cigarette smoking and intratracheal instillation of lipopolysaccharide. Six weeks later pulmonary functions were tested, bronchoalveolar fluid and arterial blood gases were assayed, and histopathological evaluations were performed in turn. The expressions of TACE, TNF-alpha and ErbB3 in lungs of all rats were determined histochemically.

RESULTSThe expressions of TACE, TNF-alpha and ErbB3 were significantly higher in group C than in group N (P < 0.01). The contents of TNF-alpha in serum (P < 0.01) and bronchoalveolar lavage fluid (BALF) (P < 0.01) were elevated more significantly in group C than in group N. A positive correlation existed between TACE and TNF-alpha (r = 0.784, P < 0.01) and between TACE and ErbB3 (r = 0.526, P < 0.01) respectively.

CONCLUSIONSTNF-alpha and ErbB3 are involved in the pathogenesis of COPD. TACE contributes to the progress of COPD indirectly through the function of TNF-alpha and ErbB3.

ADAM Proteins ; analysis ; physiology ; ADAM17 Protein ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Immunohistochemistry ; Lung ; physiopathology ; Male ; Pulmonary Disease, Chronic Obstructive ; metabolism ; pathology ; Rats ; Rats, Wistar ; Receptor, ErbB-3 ; analysis ; physiology ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo investigate the inhibitory effect of RNA interference on chronic myeloid leukemia (CML) bcr/abl oncogene expression.

METHODSThe small interference RNAs (siRNAs) were synthesized in vitro. K562 cells stably expressing bcr/abl gene were transfected with the siRNA by electroporation, both the non-transfected cells and non-specific siRNAs transfected cells were taken as controls. The enhanced green fluorescent protein (EGFP) plasmid was used as positive control and the transfection efficiency was detected by flow cytometry. Inhibitory effect of siRNAs was demonstrated by real-time quantitative RT-PCR and Western blots. Cell proliferation was measured by MTT assay and apoptosis by Annexin V-FITC assay.

RESULTSThe transfection efficiency was about 70%. The synthesized siRNAs inhibited CML bcr/abl oncogene expression at both mRNA and protein levels. siRNAs could inhibit K562 cell proliferation to 47% and 56% at 24 h and 48 h after transfection, respectively, and induce cell apoptosis from 1.00% in control group to 15.05% and 19.4% at 24 h and 48 h respectively.

CONCLUSIONAt the cell level, inhibition of CML bcr/abl oncogene expression by chemically synthesized siRNAs provides the new method for anti-leukemia study.

Apoptosis ; genetics ; Cell Proliferation ; Fusion Proteins, bcr-abl ; genetics ; Humans ; K562 Cells ; RNA, Small Interfering ; Transfection

To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B ; genetics ; metabolism ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Caco-2 Cells ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Up-Regulation ; drug effects