Objective To investigate the effect of miR-15b and miR-16 on the expression of vascular endothelial growth factor (VEGF) protein in human embryonic lung fibroblast (HLF) cells under hyperoxia.Methods The expression level of miR-15b and miR-16 was up-regulated and down-regulated in HLF cells by transfection technology, respectively.The expression of VEGF protein in HLF cells was assessed by Western blot.Furthermore, under hyperoxia exposure in vitro, the expression of miR-15b, miR-16 and VEGF protein in HLF cells was analyzed.Results Up-regulation of miR-15b and miR-16 suppressed VEGF protein expression, while down-regulated miR-15b and miR-16 promoted VEGF protein expression.In addition, hyperoxia exposure induced up-regulation of miR-15b and miR-16, but down-regulation of VEGF protein in HLF cells.Conclusions Hyperoxia exposure may up-regulate the expression level of miR-15b and miR-16, but suppress VEGF protein expression.These may contribute to the development of bronchopulmonary dysplasia.

Objective To isolate and identify the cells from the cerebral cortex,cerebellar cortex,hippocampus,sub-ventricle region,brain stem and the spinal cord of neonatal rats and observe their growth morphology in vitro.Methods The cells were incubated from different regions of the CNS of neonatal rats by using DMEM/F12 media added with 10% bovine serum,and their growth morphology was observed by using inverted phase contrast microscope;then in the 10th day after incubation cells were fixed and immunocytochemical method was used to detect specific NSE antigen of the neurons and specific GFAP antigen of the astrocytes.Results Both neurons and astrocytes were studied in each region and they bloomed in the 10th day after incubation.Neurons had big triangular or ellipsoidal cell bodies surrounded with a halo and had robust nervous process which interlaced each other around cell bodies.The astrocytes had an ellipsoidal nucleus located at one side of the cell body and they had abundant processes branching profusely.Conclusion A method of culturing cells from different regions of the CNS of neonatal rats was described.A comparative morphology study of their growth was made and neurons and astrocytes of all the regions studied were identified.

ObjectiveTo explore the effects of Jinyaodai on neurological behavior and Akt expression in the cortex of rats following spinal cord contusion injury.MethodsRats were randomly divided into control group,spinal cord contusion group and Jinyaodai group.The weight-drop device was employed to prepare the spinal cord injury(SCI) model.Jinyaodai was administrated every day by using a stomach tube.Rats were performed the BBB assessment,and the detection of Akt expression and count of Neun positive neurons in cortex following SCI.ResultsCompared with control group,deficit of motor function in hindlimbs was seen at 3 dpo following cord contusion,and partial functional recovery could be seen from 7 dpo to 1 m.Treatment of Jinyaodai greatly increased the BBB scores ( 14.1 ± 1.4 ) more than SCI group ( 7.8 ± 1.3 ) at 1 month (P ＜ 0.05 ) ; Simultaneously,compared with SCI rats,treatment of Jinyaodai significantly increased the expression of Akt (0.53 ± 0.05,0.68 ± 0.07,P ＜0.05 ) and the number of neurons ( 11 ± 2, 15 ± 1 ; P ＜ 0.05 ) in the lesion-induced cortex of rats.Conclusion Jinyaodai may play an essential roles in functional recovery after spinal cord injury,in which the underlying mechanism may be involved in the expression of Akt in cortex.

57??m).In addition,the positive product of BDNF was also observed in a few satellite cells.At 3 days after partial dorsal rhizotomy,the A value of BDNF and its mRNA in the medium and small sized neurons decreased apparently than that in normal group(P0 05).On the contrary,the A value of BDNF and its mRNA in the large sized neurons has not apparent changed at 3 days after partial dorsal rhizotomy but decreased apparently at 7 days after operation(P

Objective To explore the changes on BDNF mRNA in transected spinal cord and associated motor cortex and skeleton muscle following cord injury in rats.Methods 20 adult Sprague Dawleys rats were performed spinal cord transected operation at T11 level,then rats in each group(n=5) were sacrificed on 1,3,7 and 14 days post operation respectively.Other 5 rats were used as normal control without operation.The tissues from the rostral,caudal segments near injury site,cerebral cortex and linea obliqua tibiae were harvested.Total RNA was extracted with Trizol reagent separately.The BDNF mRNA expression in each group was detected by RT-PCR.Results(1)BDNF positive bands were seen in the tissues of the rostral,caudal segments near injury site,cerebral cortex and linea obliqua tibiae.Moreover,BDNF level in cerebral cortex is more than in the spinal cord at normal control(P

Objective To explore the engrafted effect of NSCs combined with OECs on the motor function repair of hindlimbs in rats subjected to spinal cord transected injury.Methods The NSCs and OECs were isolated from the Transgenic Green Fluorescent Protein(GFP) embryo mouse' hippocamp and neonatal mouse' olfactory bulb.Rats were divided into NSCs group,OECs group and co-transplantation group as well as control group.The recovery of posterior limb motor function in rats subjected to spinal cord transected injurywas evaluated by the BBB locomotion score respectively in each weekend among eight weeks.This was followed by a histochemiscal observation to know the engrafted cells survival and migration in the spinal cord of host under the fluorescence microscope.Results The NSCs and OECs cells survived and migrated in the spinal cord of transplantation groups:the motor functional improvement of hindlimb in rats was seen in all cell transplanted groups;NSCs group,OECs group and co-transplantation group demonstrated a significant increase than that of control group at 3 and 4 weeks post operation(P 0.05).Conclusions The engrafted NSCs,OECs and co-transplantation can improve partly the motor function recovery of hindlimb in rats at 3 to 4 weeks following cord transection;The OECs group and co-transplantation have better effects than that of NSCs group,but there is no obvious difference between the OECs group and co-transplantation group at observed time point in this investigation.

BACKGROUND:There are several routes for stem cel transplantation;however, it is stil unable to determine which one is the best. As for the different individuals with brain injury, the type of transplanted cel s, transplantation route and time wil affect the therapeutic effects.
OBJECTIVE:To investigate the effect of bone marrow mononuclear cel s transplanted via different approaches on neurological function of rats with traumatic brain injury.
METHODS:Bone marrow mononuclear cel s of rats were administered gradient centrifugation with Ficol lymphocyte separation medium, and were labeled with CFDA-SE in vitro as standby. Rat models of traumatic brain injury were established by the method of freefal . After successful establishment of rat models, bone marrow mononuclear cel s labeled with CFDA-SE were immediately transplanted into rats via injured area, lateral ventricle and internal carotid artery. One control group was designated for each transplantation route (bone marrow mononuclear cel s were replaced with the same volume of DMEM). The degree of neurological deficits was evaluated using mNSS scores at different time points after treatment. The brain tissue was harvested after the last neurobehavioral evaluation. The survival and migration of bone marrow mononuclear cel s in the injured area were observed under an inverted fluorescent microscope.
RESULTS AND CONCLUSION:At 7, 10, and 14 days after treatment, the mNSS scores of rats in al groups were al lower than those at 1 and 3 days (P<0.05). At 7 and 10 days, the mNSS scores of rats in the internal carotid artery transplantation group were significantly lower than those in the control group (P<0.05). At 14 days after treatment, the number of fluorescence-labeled cel s of rats in the internal carotid artery transplantation group was greater than that in the other groups (P<0.05) and these labeled cel s were widely distributed. The results demonstrate that the neurological function of rats can be improved by transplanting bone marrow mononuclear cel s via the internal carotid artery, and a large number of transplanted cel s can survive and migrate in the injured area.

ObjectiveTo explore the effects of hyperbaric oxygen on neurological behavior and vascular endothelial growth factor (VEGF) in rats with traumatic brain injury (TBI). MethodsThirty rats were randomly divided into three groups, ie, control group, TBI group ( a 50 g weight-drop device was employed and fell from 30 cm height to induce the injury) and hyperbaric oxygen group ( HBO group,treated with hyperbaric oxygen once per day for seven days after TBI), 10 rats per group. Neurological severity score (NSS) was used to evaluate the movement and balance impairment in all groups. Expression of VEGF was detected by means of immunocytochemical staining.ResultsAfter TBI, the rats presented different degrees of convulsions, paralysis and balance dysfunction. The NSS score was (5.6 ±1.1 ) points in the TBI group and (0.3 ± O. 1 ) in the control group, with statistical difference ( P ＜0.05). While NSS score was (3.7 ± O. 7) points in the HBO group, showing a significant decrease compared with that in the TBI group (P ＜ 0. O1 ). Immunohistochemical staining showed 15 ± 3 positive neurons of VEGF in the TBI group, significantly less than 27 ± 2 in the control group ( P ＜ 0.05 ). There were 21 ±2 positive neurons of VEGF in the HBO group, significantly less than 21 ±2 in the TBI group (P ＜0.05). Conclusion Hyperbaric oxygen may attenuate experimental traumatic brain injury by stimulating production of VEGF.

The present study aims to isolate neural stem cells from neonatal rat hippocampus and induce them to differentiate into cholinergic neurons. A multipotent cell line derived from the hippocampi of neonatal rats which had the ability to form clones was incubated in serum-free DMEM/F12 medium added with 20ng/ml basic fibroblast growth factor (bFGF) and B27. After differentiation of the neural stem cells, immunocytochemistry was used to detect nestin, the antigen of the cell clone, and β-tubulin (Tuj 1 ), glial fibrillary acidic protein (GFAP) and galactocerebroside (Galc), the markers specific for neurons, astrocytes and oligodendrocytes, respectively. Embryonic chick skeletal muscle extract was used to induce the differentiation of the neural stem cells into cholinergic neurons. The results showed that the cell line isolated from the hippocampi of neonatal rats expressed nestin and had the potential to form clones and differentiate into neurons, astrocytes and oligodendrocytes. Embryonic chick skeletal muscle extract can induce 9.6% of the isolated cell line to differentiate into cholinergic neurons compared with 3.9% in controls. These findings suggested that the cell line, which expressed nestin antigen, was a multipotent cell line capable of self-renewing, and was believed to contain stem cells of the CNS. These neural stem cells can be induced to differentiate into cholinergic neurons by using embryonic chick skeletal muscle extract.

Objective To isolate and identify the Dengue virus(DEN)from the sera of patients with unknown fever in summer and autumn in Dushan and Xingyi areas of Guizhou province,China.Methods From June 2005 to October 2005.356 blood samples were collected from patients with unknown fever in Dushan and Xingyi areas of Guizhou province.The serum samples were inoculated on the C6/36 cell monolayers.After three blind passages,the cytopathie effect(CPE)Was observed.Identification of DEN antigen was earried out by indirect immunofluorescence assay(IFA)with the monoclonal antibody(McAb)against DENl-4 virus.The total RNA was extracted from the serum and tested by RT-PCR with the universal primer for DEN NS region.And determination of the RNA sequenee Was performed,and phylogenetic analysis was carried out.Results Three serum samples caused CPE and were proved as DEN2 positive by IFA,RT-PCR and senquence determination.The phylogenetic tree analysis showed that the isolated virus strains had the closest relations with the systemic evolution of the strains DEN2-43 and DEN2-44.Conclusion DEN infections exist in the population of Dushan and Xingyi of Guizhou,China.