Objective To observe the effect of neural stem cell (NSC)transplantation on the hindlimb motor function and caspase-3 expression of motor cortex (MC) in spinal cord transected (SCT) rats. Methods S prague-Dawley (SD) rats were randomly divided into sham operated group, operation group( T9 transection), and subacute NSC transplantation group. The MC of each group( n= 8 )was harvested 7 days post operation (dpo), then western blot was employed to detect the level of caspase-3( β-actin was used as control ). The left 5 animals of each group were subjected to BBB score evaluation at 16th week,then the animals were sacrificed and the MC was harvested and performed immunostain by using caspase-3 rabbit antibody. Results Following NSC transplanta tion, BBB scores( ( 7.58 ± 0. 99 ) ) increased significantly than seen in the SCT animals( ( 5.16 ± 1.19) ). The expressional level of caspase-3 at 7day post operation was( (0.89 + 0.12)) in MC of SCT rats,while it decreased significantly to( (0.76 + 0.11 ) ) in NSC transplantation rats(P＜0.05). The immunoreactive stain of caspase-3 was seen in the cytoplasm of pyramidal neuron in the cortex. Conclusion NSC engraft can downregulate the expression of caspase-3, corresponding to a significant improvement in hindlimb motor function. These findings indicate that NSC transplantation probably regulate the expression of apoptosis genes in MC to promote neurological function recovery in rats subjected SCT.

The present study investigated different types of eoexpression of brain derived neumtmphic factor(BDNF),nerve growth factor(NGF)and neutmphin-3(NT-3)mRNA and/or proteins in the left sixth lumbar dorsal root ganglion(DRG)of cats and discuss themechanism of coexpression in order to provide foundation for elucidating the relationship between the expression of neurotrophic factors andspinal cord plasticity.The eats used in this study were normal animals without any interventional treatment.They were subjected to renloveof the left L6 DRG and their DRG were processed for immunohistechemistry and in situ hybridization double staining to observe whetherthere are coexpression of mRNA and proteins of BDNF,NGF and NT-3.The results showed that the pmteios and mRNA of BDNF,NGFand NT-3 were all expressed in the DRG of cats,but the types of coexpression of mRNA and proteins were different and diverse amongthese three neumtrophic factors.The results of immunohiatochemistry showed that BDNF immunoreactivities were mainly observed in thecytoplasm and nucleus,and the staining of nucleus was weaker than that of cytoplasm;NGF immunoreactivities were mainly observed innucleus while NT-3 mainly in cytoplasm.The results of in situ hybridization showed that BDNF and NGF positive signals mostly distributedin the cytoplasm,NT-3 positive signals were observed in both the cytoplasm and nucleus.Our results suggest that the proteins and mRNAof BDNF,NGF and NT-3 have different types of coexpression which indicate they may have autocrine and/or paracrine mechanism contrib-uting to the plasticity of spinal cord in the left L6 DRG of cats.

BACKGROUND: Many studies showed that neural stem cells (NSC) transplantation can promote functional improvements in rats subjected to spinal cord injury. However, the underlying molecular mechanisms remain poorly understood.OBJECTIVE: To observe the effects of NSC transplantation on expressions of Bax, Bcl-2 and Caspase-3 in the motor cortex in rats subjected to spinal cord transection.DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed at Institute of Neuroscience,Kunming Medical College from July 2007 to December 2008.MATERIALS: Five green fluorescent protein transgenic mice with 14-15 embryonic days were prepared for NSC. Additionally 88 adult female Sprague-Dawley (SD) rats were randomly divided into sham operation group (n=8), model group (n=40) and NSC transplantation group (n=40).METHODS: Model of spinal cord transection was established by cut transversely Sprague-Dawley (SD) rats T_9 segment. Rats in the sham operation group were subjected to laminectomy at T_8, without spinal cord injury. After the spinal cord was exposed at the lesion site, a small piece of 2 mm~3 gel foam soaked with 3×10~5 NSC were implanted into the gap to fill the lesion site of the T_9 level in rats of the NSC transplantation group. The measurements of relative indexes were performed at the days 3, 7,14, 21 and 28 after transplantation.MAIN OUTCOME MEASURES: Changes of Bax, Bcl-2 and Caspase-3 expressions were detected by RT-PCR.RESULTS: Compared to the sham operation group, the Bax expression in the model group had no significant difference (P >0.05), the expression of Bcl-2 was decreased obviously at days 14 and 28 after operation (P < 0.05), while a significant increase on the expression of Caspase-3 at day 3 (P < 0.05). Compared to the model group, the expression of Bax was dramatically decreased in the NSC transplantation group at day 3 (P < 0.05), with a notably increased Bcl-2 expression at days 14 and 21 after transplantation (P < 0.05), but the expression of Caspase-3 presented a significant decrease at days 3 and 7 after transplantation (P < 0.05).CONCLUSION: NSC transplantation probably regulates the expression of apoptosis genes (Bax, Bcl-2 and Caspase-3 mRNA) to promote neurological function recovery in rats subjected to cord transection.

Objective To observe the clinical effects of Zaobo decoction combined bisoprolol treating premature ventricular contractions.Methods 108 patients of ventricular premature beat were recruited intoa control group and an observation group (n=54) according to the random number table method.The control group was treated with bisoprolol, while the observation group was treated with Zaobo decoction on the basis of the control group, and both groups were treated for 8 weeks.24 h dynamic electrocardiogram (ECG), renin activity plasma (PRA), angiotensin Ⅱ (angiotensionⅡ) and ALD (Ang) were observed before and after treatment.The clinical effects were evaluated.Results The total effective rate showed significant difference between the observation group and the control group (75.9％ vs.57.4％;x2=4.167, P=0.041) after the treatment.After treatment, Ang-Ⅱ (56.22 ± 12.7 pg/ml vs.68.45 ± 12.7 pg/ml, t=5.004) in the observation group was significantly lower than the control group (P＜0.01);24 h sinus RR interval standard deviation (129.16 ± 28.56 ms vs.116.13 ± 17.38 ms, t=2.864), every 5 min sinus RR interval mean standard deviation within 24 h (123.57 ± 25.24 ms vs.112.46 ± 18.23 ms, t=2.622), and within 24 h of sinus RR interval difference rms (31.76 ± 11.42 ms vs.22.64 ± 10.32 ms, t=4.354) in the observation group were significantly higher than the control group (P＜0.01).Conclusion Zaobo decoction combined with bisoprolol can effectively improve heart rate variability, regulate rernin vascular angiotensin system, and improve the clinical efficacy of the patients with ventricular premature beat.

Objective To explore the efficacy of low tidal volume ventilation strategy in children with acute hypoxia respiratory failure (AHRF).Methods A total of 79 hospitalized children with AHRF from Aug 2006 to Jul 2011 in PICU of Kunming Children's Hospital were enrolled in this study.The observation group in-cluded 55 children who received low tidal volume ventilation strategy (6-8 ml /kg),while the other 24 children (control group)were given traditional mechanical ventilation (10-12 ml /kg).Oxygenation situations such as PaO2 ,PaCO2 ,PaO2 /FiO2 ,oxygen index and blood gas pH value,organ function,mechanical ventilation complica-tions,hospitalization days and expenses in PICU and the mortality were observed.Results (1)PaO2 ,PaO2 /FiO2 and oxygen index in the observation group were better than those in control group after 24 h mechanical ventilation [(68.51 ±7.53)mmHg(1 mmHg =0.133 kPa)vs.(61.64 ±9.28)mmHg,(162.9 ±21.84)mmHg vs.(152.1 ± 19.03)mmHg,and 18.85 ±4.1 vs.26.53 ±5.2,respectively],and there were significant differences between two groups (P ﹤0.05);and there were also significant differences between two groups in the results after 48 h and 72 h mechanical ventilation.(2)The PaCO2 was (47.48 ±10.52)mmHg after 24 h in observation group,while the PaCO2 in control group was (30.17 ±6.59)mmHg,and it suggested excessive ventilation.(3)Mechanical venti-lation time (7.6 ±3.1)d and hospitalization days (12.8 ±3.6)d were shorter in observation group(P ﹤0.01). Barotrauma (7.3%)and mortality (20.0%)in observation group was significantly lower than those in control group (29.2%,41.6%;P ﹤0.01).The number of damaged organs in observation group was lower than that in control group (P ﹤0.05).Conclusion Low tidal volume ventilation with appropriate positive end expiratory pressure could improve oxygenation,prevent alveolar collapse,reduce complications and mortality for children with AHRF,it should be applied for the treatment of children with AHRF.

Objective To establish a serial of stable and mature methods of primary culture hippocampus neurons of GFP-transgene embryonic mice,to get the morphologic data of cultured neurons.Based on this research,in future,we can give an important theoretical and practical support for the therapy of nervous system diseases by transplanting with hippocampus neurons of GFP-transgene embryonic mice.Methods We primarily cultured the hippocampus neurons derived from the GFP-transgene embryonic mice in vitro.Under the microscope,we found cultured hippocampus neurons could live for more than one month and appeared to be the best status in 5～7 d after culture.During this time,the processes of the neurons are thick and the neurons connected one another to form the"cells-net" through their processes.After 14 d,the growth of the hippocampus neurons became slow.Results A serial of culture methods of hippocampus neurons had been successfully established.These cultured neurons were identified by the immuno-histochemical methods.They grow well in different phases before 14 d after culture.Conclusion Culturing hippocampus neurons of GFP-transgene embryonic mice is a simple,stable and effective method which can be applied to scientific research by other researchers.

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 effect and injury mechanism of reactive oxygen species (ROS) after spinal cord injury (SCI) through detecting the dynamic changes of malonyldialdehyed (MDA)content in spinal cord and observing neurocyte apoptosis and correlation apoptosis factor expression after SCI. Methods Totally 132 adult SD male rats were randomly divided into three groups: sham group, SCI group, methylprednisolone (MPSS) group. The SCI of SD rats was performed by Allen's weight dropping way to impact on the posteriors of spinal cord T_(10). The contents of MDA were determined by chromatometry, the expression of Caspase-3 and Bcl-2 family in the injured spinal cord was detected by immunohistochemical staining;Apoptotic cells were detected by using fluorometric terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (fluorometric TUNEL) staining. Results The content of MDA in the injured cord increased significantly after SCI;R3eached the peak at 6 hours and 3 days post-injury, then dropped down gradually, then was back to the normal level after 7 days. The number of TUNEL labeling positive cells of SCI group increased at 6 hours post-injury;R3eached the peak at 3 days, then dropped down gradually;Bcl-2, Bax protein began to increase at 6 hours post-injury;R3eached the peak at 5 days after injury, then dropped down gradually. Caspase-3 protein began to increase at 6 hours post-injury;R3eached the peak at 3 days after injury, then dropped down gradually. The content of MDA, the number of TUNEL labeling positive cells, the expression of Caspase-3 and Bax of MPSS group decreased significantly than that of SCI group at the same time;R3espectively, while Bcl-2 protein was up-regulated after administration of MPSS.Conclusion ROS could promote the expression of Caspase-3 and degrade the ratio of Bcl-2/Bax to induce apoptosis of neurocyte, which might play significantly role in the process of secondary SCI. In addition, MPSS exerts neuroprotective effects against ROS toxicity, which might be of importance and might contribute to their clinical efficacy for the treatment of SCI.

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.

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.