Effect of chondroitin sulfate enzyme ABC on glial scar in brain injury models / 中华神经医学杂志

Objective To explore the effect of chondroitin sulfate enzyme ABC (chABC) on glial scar in rat models of brain traumatic injury (TBI). Methods Thirty-eight Wistar rats were randomly divided into 5 groups, including normal control group (n=2), model group (rat models of TBI,n=9), 1.0 U/mL chABC treatment group (n=9), 2.5 U/ml chABC treatment group (n=9) and 5.0 U/ml chABC treatment group (n=9). After performing TBI by free falling in the later 4 groups, rats of the model group were given no treatment, while those of the other 3 groups were administrated with different concentrations of chABC by local injection respectively. One, 2 and 4 w after TBI, HE staining was performed on the brain tissues of these rat models;and immunohistochemical assay and Western blotting were employed to evaluate the secreting of chondroitin sulfate proteoglycans (CSPGs) and the therapeutic effect of chABC on glial scar. Data were statistically analyzed using t-test. Results Pathological test revealed the scars in the treatment groups were significantly fewer than those in the model group 2 w after TBI, with 5.0 U/mL chABC treatment group enjoying the fewest level (P<0.05). Immunohistochemical assay showed that the secreting of CSPGs in the treatment groups and model group was significantly increased than that in normal control group 2 w after TBI (P<0.05);the 5.0 U/ml chABC treatment group showed an obvious reduction of CSPGs secreting as compared with the model group (P<0.05). Western blotting indicated that the treatment groups showed an obvious reduction of CSPGs secreting as compared with the model group 1, 2 and 4 w after TBI (P<0.05);an obvious gradual reduction of CSPGs secreting in the model group, 2.5 and 5.0 U/ml chABC treatment groups was noted 1, 2 and 4 w after TBI (P<0.05). Conclusion ChABC could degrade the glial scar by degrading the CSPGs molecules and improve the microenvironment of local axonal regeneration after TBI;In this experiment, the highest concentration of chABC (5U/ml) shows the best effect on removing the glial scar.

Construction of a mouse pDsRed2-N1-SDF-1alpha eukaryotic expression vector and its expression in mouse bone marrow mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector pDsRed2-N1-SDF-1alpha and observe its expression in the mouse bone marrow mesenchymal stem cells.</p><p><b>METHOD</b>SDF-1alpha gene sequence with XhoI, EcoRI restriction enzyme cutting site was amplified from the total RNA of mouse smooth muscle cells by reverse transcription-polymerase chain reaction (RT-PCR) and inserted into the eukaryotic expression vector pDsRed2-N1 encoding red fluorescent protein gene, and the insertion was verified by endonuclease digestion and DNA sequencing. Mouse bone marrow mesenchymal stem cells identified with immunofluorescence assay for vimentin expression were transfected with the constructed plasmid pDsRed2-N1-SDF-1alpha, and the expression of sdf-1alpha was detected using immunofluorescence assay.</p><p><b>RESULTS</b>The DNA fragment amplified by PCR from the total RNA was identical to SDF-1alpha from the gene library, and an identical DNA fragment was also amplified from the recombinants. Sequence analysis confirmed the successful insertion of SDF-1alpha into the pDsRed2-N1 vector and the eukaryotic expression vector pDsRed2-N1-SDF-1alpha was successfully constructed. The cultured mouse bone marrow mesenchymal stem cells positive for vimentin protein showed SDF-1alpha expression 24 h after transfection with the recombinant vector.</p><p><b>CONCLUSION</b>The pDsRed2-N1-SDF-1alpha eukaryotic expression vector constructed is capable of expression of SDF-1alpha fusion protein in the mouse bone marrow mesenchymal stem cells.</p>

Immunogenicity of recombinant DNA vaccine that coded for the neurite growth inhibitors / 中华神经医学杂志

Objective To detect the immunogenicity of the recombinant DNA vaccine that encoded for neurite growth inhibitors: Nogo-A, oligodendrocyte myelin glycoprotein (OMgp), tenascin-R (TN-R) and myelin-associated glycoprotein (MAG) after the nerve injury under the help of pAdEasy, a kind of adenovirus plasmid being the vector of the DNA. Methods Sixteen 5-w-old Lewis rats were randomized into DNA vaccination group (vaccine group) and pAdEasy group. Rats in the vaccine group were immunized once weekly for a consecutive 8 w by bilateral injection of the recombinant plasmid into the musculus tibialis. The immunized animals in the 2 groups were exsanguinated each time before the vaccination for sera collection, and the qualitation and quantitation of the antibodies in the serum were detected by Dot-blot analysis and ELISA. Results The vaccine group could produce fusion-protein antibodies against Nogo-A, MAG, OMgp and TN-R at the 6th w of vaccine injection, while pAdEasy group could not. The valency of antiserum was shown by ELISA as 1:1 000 000 at the 6th w of vaccine injection and kept this level stably. Conclusion The DNA vaccine exclusively induces the generation of the fusion-protein antibodies against Nogo-A, MAG, OMgp and TN-R in vivo, which controls the favorable immunogenicity.

Analysis of the trace elements in the hair of elderly patients with Alzheimer's disease / 中华神经医学杂志

Objective To investigate the association between the trace elements and senile dementia.Methods The trace elements including zinc,iron,copper,manganese,and lead elements and calcium in the hair of the elderly patients with Alzheimer' s disease(AD)admitted in our hospital from January,2006 to September,2008 were analyzed,and the results were compared with those from 46 control subjects.Results Zinc,calcium and manganese levels in the AD group were significantly lower than those in the control group(P＜0.05).Conclusion Lowered contents of the trace elements zinc,calcium and manganese might be associated with the pathology of AD.