Expression and its significance of vascular endothelial growth factor in patients with obstructive sleep apnea-hypopnea syndrome / 中华老年医学杂志

Objective To observe the expression level change of vascular endothelial growth factor (VEGF) in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS), and to explore the relationships of VEGF expression with OSAHS, OSAHS related cardiovascular and cerebrovascular diseases. Methods Polysomnography (PSG) was used to conduct sleep apnea monitoring in 24 OSAHS patients from 6OSAHS popular families and 48 healthy controls with normal physical examination results. The expression of VEGF mRNA was examined by reverse transcriptionpolymerase chain reaction (RT-PCR) analysis, meanwhile, the level of VEGF in plasma was measured VEGF mRNA in PBMC were significantly higher in simple OSAHS group [plasma levels: (205.75±2.79) pg/ml; mRNA: 0. 61±0. 02] than in control group [(168.72±4.64) pg/ml; 0. 47±0. 02,cardiovascular and cerebrovascular diseases group [(288.74 ± 2.73) pg/ml, 1.16 ± 0. 03] than in simple OSAHS group [ ( 205.75 ± 2.79 ) pg/ml, 0. 61 ± 0.02, P ＜ 0. 01]. ( 2 ) There was a positive correlation of the levels of VEGF in plasma and mRNA with AHI as well as systolic and diastolic blood pressure in early morning. There was a negative correlation of the level of VEGF in plasma and VEGF mRNA with the lowest saturation of blood oxygen. There was a positive correlation of the level of VEGF mRNA with AHI as well as systolic and diastolic blood pressure in early morning.Conclusions The level of VEGF in OSAHS significantly increases, which may play a role in the pathophysiology of OSAHS and OSAHS related cardiovascular and cerebrovascular diseases.

Therapeutic efficacy of lentiviral vector mediated BDNF gene-modified MSCs in cerebral infarction / 生物工程学报

Pretreatment with brain-derived neurotrophic factor (BDNF) reduces ischemic damage after focal cerebral ischemia, and bone marrow mesenchymal stem cells(MSCs) were reported to ameliorate functional deficits after stroke in rats. Here we investigate the synergistically therapeutic effects of BDNF gene-modified MSCs on cerebral infarction. We transfected MSCs with the BDNF gene using a lentivirus-based system and investigated whether the BDNF-modified MSCs contributed to improved functional recovery in a rat transient middle cerebral artery occlusion (MCAO) model. Compared to untreated rats, rats that received both MSCs and BDNF-MSCs showed significantly more functional recovery. The difference in modified neurological severity score(mNSS) was statistically significant (P < 0.001). Recovery was better in BDNF-MSCs than in MSCs (P < 0.001). At the second week and second month after the systemic delivery of blank vector-modified MSCs and BDNF-modified MSCs, the treated rats exhibited more significant recovery than the control, including the accumulation and living of enhanced green fluorescence protein (EGFP)-positive cells in the infarct area and surrounding areas, neuron-like changes, expression of surface markers of neural cells, and a large amount of BDNF expression in the BDNF-MSCs-treated group. Our findings suggest that BDNF-gene-modified rMSCs can migrate to surrounding areas of the cerebral infarction lesion, differentiate into neural cells, and survive for extended periods. With the synergy of BDNF, they may promote the recovery of the neurological function following cerebral infarction and represent a new strategy for stem cell-based therapy.

Brain-derived neurotrophic factor gene-modified bone marrow mesenchymal stem cells in treatment of sciatic nerve injury / 中国组织工程研究

BACKGROUND: Peripheral nerve injuries are hoped to promote the regeneration of nerve repair by elevating brain-derived neurotrophic factor (BDNF) concentration in local injury regions. OBJECTIVE: To observe the recanalization of nerve fiber and motor function in sciatic nerve injury rats following BDNF gene-modified bone marrow mesenchymal stem cells (BMSCs). DESIGN, TIME AND SETTING: The randomized, controlled animal study was performed at the Fujian Institute of Neurology from May 2007 to May 2008. MATERIALS: Femur and tibia of F344 male rats aged 2 months were sterilely harvested to prepare BMSCs. BDNF gene-modified BMSCs were prepared using constructed chronic viral vector PNL-BDNF-IRES2-EGFP. METHODS: The right sciatic nerve injury models were made using 60 adult Sprague Dawley rats. All models were randomly assigned into phosphate buffer saline (PBS) group, BMSC group and BDNF gene-modified BMSC group. 2 ?L BPS, 2 ?L BMSC solution and 2 ?L BDNF modified BMSC solution were separately transfered into injury site of each group. MAIN OUTCOME MEASURES: The retrograde horseradish peroxidase tracing was practiced to observe neural cell number in the Lumbar 4 and 5 spinal cord anterior horn at 2 and 4 weeks after microinjection. Sciatic nerve function index was used to observe rat motor function of injured limbs. Fluorescence excitation and immunohistochemistry were employed to detect BMSC survival and BDNF expression. RESULTS: Cell number was more, and nerve function recovery was better in the Lumbar 4 and 5 spinal cord anterior horn in the BDNF gene-modified BMSC group than in the PBS and BMSC groups. BMSC survival was found in the injured sites in the BMSC and BDNF gene-modified BMSC groups. BDNF expression was significantly more in the BDNF gene-modified BMSC group than in the BMSC group. CONCLUSION: BDNF gene-modified BMSCs have promotion effects on the recanalization and functional recovery of nerve fiber following peripheral nerve injury.

Construction of lentiviral vector carrying the Ang-1 gene and its expression in the rMSCs / 中国病理生理杂志

AIM: To construct lentiviral vector carrying the angiopoietin-1(Ang-1) gene,and make it express Ang-1 in the rat mesenchymal stem cells(rMSCs).METHODS: The cDNA encoding the CDS of Ang-1 gene was obtained from the placenta of the adult Fisher 344 rats with RT-PCR.After digestion with restrication endonuclease,the Ang-1 gene was recombined to construct the transfer plasmid PNL-Ang1-IRES2-EGFP.The three-plasmid system of lentiviral vector was consisted of PNL-Ang1-IRES2-EGFP,the packaging plasmid HELPER,and the envelope plasmid VSVG,which were co-transfected to 293T cells mediated by lipofectamin2000 to produce lentiviral particles.The rMSCs were infected by obtained lentiviral particles.The insertion of Ang-1 gene was detected by PCR,the mRNA expression of Ang-1 in rMSCs was detected with RT-PCR,the protein expression of Ang-1 was observed with immunocytochemistry and Western blotting methods.RESULTS: The result of sequencing showed that the cloned Ang-1 gene was consistent with the sequence reported in GenBank.After digestion with restrication endonuclease,the 1 512 bp fragment of Ang-1 gene and the 10.5 kb vector fragment of PNL-IRES2-EGFP were observed with gel electrophoresis.The insertion of Ang-1 gene in viral genome was confirmed.The EGFP expression was observed with the fluorescent microscope.In infected rMSCs,the mRNA and protein expressions of Ang-1 were confirmed.CONCLUSION: Lentiviral vector carrying Ang-1 gene has been successfully constructed.The infected rMSCs are able to express the Ang-1 mRNA and Ang-1 abundantly.This will facilitate the following exploratory development of Ang-1 gene-modified rMSCs.