Isolation and multipotent differentiation of human decidua basalis-derived mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the biological features of human decidua basalis-derived mesenchymal stem cells (PDB-MSCs) in vitro and identify their capacity of multilineage differentiation.</p><p><b>METHODS</b>PDB-MSCs were harvested from the decidua basalis of term placental by enzymatic digestion and density gradient centrifugation, and the growth characteristics and morphological changes of the MSCs were observed by inverted microscope. The proliferative ability of the cells was assessed by Cell Counting Kit-8. The cell cycle and expressions of the surface markers (CD29, CD44, CD73, CD90, CD34, CD45, and CD14) of the MSCs were identified by flow cytometry. Multilineage differentiation capacity of the cells was tested by inducing their differentiation toward osteoblasts, adipocytes and chondroblasts in vitro.</p><p><b>RESULTS</b>MSCs isolated from human decidua basalis of term placental exhibited a morphology similar to that of bone marrow-derived MSCs, and grew into colonies in in vitro culture, where the cells proliferated rapidly after passage with a cell doubling time of 2.21∓0.21 days. More than 70% of the cells stayed in the resting stage (G(0)/G(1)) and showed positivity for CD29, CD44, CD73 and CD90, but not for CD14, CD34 or CD45. After induction, the cells showed positive results of alizarin red staining, oil red O staining and Alcian blue staining.</p><p><b>CONCLUSION</b>Human decidua basalis contains a rich source of MSCs, which can be easily isolated and cultured without affecting their capacity of multilineage differentiation. The PDB-MSCs may have the potential as a novel source of stem cells.</p>

Interventional treatment of dural arteriovenous fistula / 中华神经医学杂志

Objective To explore the clinical skills and curative efficacy of endovascular embolization on dural arteriovenous fistula (DAVF). Methods A total of 50 patients with DAVF,admitted to our hospital from 2000 to 2009, underwent endovascular embolization with different materials through a microcatheter under DSA. After embolization, imaging examination was applied and clinical manifestations were noted during the clinical follow-up to evaluate the clinical efficacies.Results Immediate postoperative angiography indicated that the fistulas were totally occluded in 43patients, occluded evidently in 6 and occluded partially in 1. The 6-84 months follow-up found that clinical symptoms of 34 patients disappeared, 6 alleviated evidently, 7 relapsed, and 3 worsened; no complications were noted. Conclusion The endovascular embolization for DAVF is effective; and choosing the right embolic approach and material and improving the technology are the keys of successful treatment.

Isolation of human umbilical cord mesenchymal stem cells and their differentiation into dopaminergic neurons / 中华神经医学杂志

Objective To investigate the methods of isolation of human umbilical cord mesenchymal stem cells (MSCs) in Wharton' s jelly and the differentiation of MSCs into dopaminergic neurons. Methods The umbilical cord mesenchymal tissue was scraped off from the Wharton's jelly,and then, collagenase Ⅳ was employed to isolate the MSCs. The isolated cells were primarily cultured in DMEM/F12 medium containing 10% FBS. Inverted microscopy was used to observe the cytomorphology, and flow cytometry was employed to detect the cell surface antigens and the cell cycle.We evaluated the cell viability using CCK8 kit. Two-step method was employed to induce the MSCs of the P3 generation to differentiate into dopaminergic neurons, and immunocytochemistry and Western blotting were used to detect the expressions of neuron specific enolase (NSE) and tyrosine hydroxylase (TH) 3, 6 and 9 d after the induction. Results The isolated MSCs showed fibroblast-like shape, with parallel arrangement and vortex-like growth. MSCs of the P3 generation expressed CD73, CD29, CD44and CD105, but did not express CD34, CD45, CD106 and HLA-DR. The doubling time in the exponential phase was at the 48th h of culture, and 91.13% cells were under G0-G1. These cells had similar morphology of the neurons. The immunocytochemical assay showed that the NSE and TH positive rates were 19.5% and 8.9% on the 9th d of induction; and Western blotting showed that MSCs obviously expressed NSE and weakly expressed TH.Conclusion MSCs can be isolated from the umbilical cord mesenchymal tissue, and be induced to differentiate into dopaminergic neurons in vitro.

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>

Determination of SARS-CoV by simplified nested fluorescent RT-PCR / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish a simple rapid and sensitive nested RT-PCR method for detection of SARS coronavirus RNA by designing the specific primers for SARS and optimizing the parameters for PCR.</p><p><b>METHODS</b>Primers and fluorescent probes were designed according to the sequences of SARS coronavirus genes available from GenBank. The optimization of the parameters for PCR was performed in PE 7700 thermal cycle. The 36 serum samples and 40 mouthwash of SARS patients and 80 samples of healthy people were tested.</p><p><b>RESULTS</b>The positive rate of patient serum and mouthwash was 33.6%, (12/36) and 67.5%, (27/40), respectively, while the positive rate of healthy people was zero (0/160).</p><p><b>CONCLUSION</b>The simple nested RT-PCR method was a rapid, efficient and sensitive one for SARS early diagnosis.</p>

Development of an intelligent control system for radiography / 中国医疗器械杂志

An intelligent control system has been designed using the single chip and the related circuit, and with the assemble language. It is connected with the common X-ray units to control the exposure dose. The result shows that three parameters for radiography are well controlled by the intelligent control system, and auto-radiography is realized.

Coxsackie virus B types were discriminated by RT-PCR / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To develop a method for detection of coxsackie B virus type 1-6 by RT-PCR.</p><p><b>METHODS</b>A pair of primers were designed to amplify all types of coxsackie B virus 1-6 efficiently. The PCR product was hybridized in micro-wells in which 6 type specific oligonucleotide probes had been coated respectively, colorimetric detection was performed to discriminate the types of coxsackie B virus.</p><p><b>RESULTS</b>This method was shown to be concordant with the IgM ELISA, 71.7% of anti-coxsackie B positive cases could be detected by RT-PCR.</p><p><b>CONCLUSION</b>The RT-PCR method can type coxsackie B virus efficiently and provides a tool for clinical diagnosis and epidemiological investigation.</p>