The experimental study of rAAV2 transfecting neural stem cells-derived neurospheres / 中国应用生理学杂志

<p><b>AIM</b>To investigate the abilities of recombinant adeno-associated virus type 2 (rAAV2) transfecting neurospheres.</p><p><b>METHODS</b>The rAAV2 conjugated with FITC (rAAV2-FITC) was added into the culture medium of neurospheres and 30 minutes later the neurospheres were detected with a fluorescence microscopy to determine if the AAV can combine with neurospheres. The rAAV2 containing GFP reporter gene (rAAV2-GFP) was incubated with the neurospheres for a month and then detected the ability of transfecting neurospheres. The neurospheres transfected with rAAV2-containing GFP were transplanted to the brain of rats. A month later the rats were sacrificed and the brains were removed to detect if there are expressions of the reporter gene. The neurospheres were transfected with rAAV2 containing hypoxia responds elements (HRE) and vascular endothelium growth factor(VEGF) gene and reporter gene GFP (rAAV2-HRE-VEGF-GFP) and then cultured in low oxygen density environments. Seventy-two hours later the neurospheres were detected through a fluorescence microscopy.</p><p><b>RESULTS</b>The neurospheres incubated with rAAV2-FITC present bright green fluorescence. GFP, the reporter gene, can be seen clearly 1 month after being transfected with rAAV2-GFP. The same green fluorescence protein can be observed ex vivo as well. The fluorescence can be seen in neurospheres transfected by rAAV2-HREVEGF-GFP only in low oxygen density.</p><p><b>CONCLUSION</b>The rAAV2 can transfect neurospheres specifically and efficiently. Reporter gene can be expressed in the neurospheres in vivo and ex vivo. Expression of reporter gene can be adjusted by HRE.</p>

Experimental study on the proliferation and plasticity of neural stem cells in situ in adult rats after cerebral infarction / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the proliferation and plasticity of neural stem cells in situ in adult rats after cerebral infarction.</p><p><b>METHODS</b>Cerebral infarction models of rats were made and the dynamic expression of bromodeoxyuridine (BrdU) and BrdU/polysialylated neural cell adhesion molecule (PSA-NCAM) were determined by immunohistochemistry and immunofluorescence staining.</p><p><b>RESULTS</b>Compared with the controls, the number of BrdU-positive cells in the subventricular zone (SVZ) and hippocampus increased strikingly at day 1 (P < 0.05), reached maximum at day 7, and decreased markedly at day 14, but it was still elevated compared with that of the controls (P < 0.05); The number of BrdU-labeled with PSA-NCAM-positive cells increased strikingly at day 7 (P < 0.05), reached maximum at day 14, and markedly decreased at day 28, but it was still elevated compared with that of the controls (P < 0.05), and was equal to 60% of the number of BrdU-positive cells in the same period.</p><p><b>CONCLUSIONS</b>Our results indicate that cerebral infarction stimulate the proliferation of inherent neural stem cells in situ and most proliferated neural stem cells represent neural plasticity.</p>

Measurement of soluble CD44v6 in peripheral blood as assistant diagnosis of invasive pituitary adenomas / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the value of measuring the concentration of soluble CD44 splice variant 6 (sCD44v6) in peripheral blood in patients with invasive and non-invasive pituitary adenomas.</p><p><b>METHODS</b>The concentrations of sCD44v6 in peripheral blood were measured with ELISA in 68 patients with invasive pituitary adenomas and 100 patients with non-invasive pituitary adenomas.</p><p><b>RESULTS</b>The serum concentration of sCD44v6 in patients with invasive pituitary adenomas was lower than that in patients with non-invasive pituitary adenomas, while the latter was lower than that in healthy controls. The serum concentrations of sCD44v6 were (44.63 +/- 7.21), (34.53 +/- 6.41), and (26.34 +/- 4.95) ng/ml in patients with invasive microadenoma, macroadenoma, and giant adenoma, and (60.78 +/- 9.61), (57.78 +/- 10.00), and (37.22 +/- 5.17) ng/ml in patients with non-invasive microadenoma, macroadenoma, and giant adenoma, lower than that in the healthy control group (68.73 +/- 6.00) ng/ml. Significant differences were observed among groups (P < 0.005). The concentration of sCD44v6 in peripheral blood decreased as the tumor size increased (P < 0.01), which was particularly significant in invasive pituitary adenomas. The positive rate in the patients with invasive pituitary adenomas reached 89.71%.</p><p><b>CONCLUSION</b>Serum concentration of sCD44v6 in the peripheral blood is inversely correlated with tumor size and its invasive growth, which may provide certain value in the early diagnosis, treatment and prognosis of invasive pituitary macroadenoma and giant adenoma.</p>