Bone marrow mesenchymal stem cells(BMSCs) are a population of multipotent cells. Many researchers have studied whether BMSCs could differentiate into neurons in vivo and ex vivo. Although some studies didn't agree with about some chemical induction medium inducing neurons, the application of cell transplantion and gene therapy of BMSCs has achieved certain progress showing good future for BMSCs in therapy of nervous system diseases.

BACKGROUND:Previous in vitro studies mainly focused on morphological and nerve marker aspects in the study of bone marrow mesenchymal stem cells (BMSCs)-derived neuron-like cells,but less focused on the olectrophysiological properties of neuron-like cells following differentiation.OBJECTIVE:To study the electrophysiological changes of differentiation from BMSCs into neuron-like cells after induction by brain-derived neurotrophic factor (BDNF)/basic fibroblast growth factor (bFGF)/alltrans-retinolc acid (AT-RA).DESIGN,TIME AND SETTING:The cytological comparative in vitro study was performed at the Department of Neural Cell Laboratory,Tianjin Huanhu Hospital and College of life Science of Nankai University from June 2005 to October 2007.MATERIALS:Totally 3 male Wistar rats (6-week old,weighing about 160 g) were used in this study.METHODS:BMSCs were cultured and purified by their characteristic of plastic adhesion in vitro,and then induced by BDNF,bFGF and AT-RA,and differentiate into neuron-like cells.Whole-cell patch damp technique was used to detect cell membrane current prior to and 3 days following induction.MAIN OUTCOME MEASURES:MSC phenotype was determined by flow cytometry.Cell morphology was observed under the inverted microscope before and after differentiation.Neuron specific enolase expression was assessed by immunocytochemistry.Whole-cell current results were measured.RESULTS:Flow cytometry results demonstrated that CD90 positive rate (99±3)%,CD31 (3.4±0.8)%,and CD34 (0.3±0.1)%.This indicated that these cells were undifferentiated stem cells,with purity of 95%.Undifferentiated MSCs under the optical microscope were mostly fiat cells with processes,similar to fibroblast-like cells.Neuron-like cells appeared 3 days following induction.Immunocytochemistry results showed that MSCs before induction were weakly positive for neuron specific enolase,but strongly positive for neuron specific enolase.At 72 hours,the differentiated rate was (24.01±3.76)%.The peak currents of outward currents in neuron-like cells were significantly higher in the induction group compared with the control group (P ＜ 0.05),but no inward Na~+ current was detected.CONCLUSION:(bFGF & BDNF)/AT-RA could induce the differentiation of MSCs into neuron-like cells.These cells showed the tendency to differentiate into mature neurons,though having no electrophysiological properties of mature neurons.

BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into osteoblasts under inducing condition that Zouguiwan and Youguiwan coordinate inducers, but the mechanism remains to be discussed. OBJECTIVE:To observe the effects of serum containing Zuoguiwan and Youguiwan on transforming growth factorβ1 and its signal transduction protein Smad2/3 message expression during the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS:A whole bone marrow adherence method was adopted to isolate and cultivate bone marrow mesenchymal stem cells from rats. The cellcultivation was processed in five groups:bone marrow mesenchymal stem cells were respectively cultured with blank serum, serum containing Zouguiwan, serum containing Youguiwan, positive serum containing progynova+inducer (dexamethasone, vitamin C, andβ-glycerophosphate), and inducer. Western blot was applied to detect the expression of type I col agen. The immunohistochemical assay was utilized to test transforming growth factorβ1 and Smad2/3 expression in the osteoblasts. RESULTS AND CONCLUSION:It was apparently more significant for serum containing Zuoguiwan and Youguiwan on type I col agen, transforming growth factorβ1 and Smad2/3 expression, compared with blank serum group and inducer group (P<0.05);moreover, serum containing Zuoguiwan was better than serum containing Youguiwan (P<0.05). Both of serum containing Zuoguiwan and Youguiwan are able to promote osteogenic differentiation of bone marrow mesenchymal stem cells. Moreover, Zuoguiwan is much more effective indicating that this method of traditional Chinese medicine about nourishing kidneys can be better to promote osteogenic induction of bone marrow mesenchymal stem cells.

The present study was performed to investigate the seroprevalence and risk factors for Dirofilaria immitis infection in cats from Liaoning province, northeastern China. From October 2014 to September 2016, sera of 651 cats, including 364 domestic cats and 287 feral cats (332 females and 319 males) were assessed. They were tested for the presence of D. immitis antigen using SNAP Heartworm RT test kit. In this population, the average prevalence was 4.5%. Age and rearing conditions (feral or domestic) were found to be associated with the prevalence of D. immitis. The prevalence was significantly higher in feral cats compared with domestic cats (8.4% vs 1.4%, P < 0.01). There was no significant difference between males and females (4.7% vs 4.2%, P>0.05), but older cats (≥3 years old) showed a statistically higher prevalence compared with younger cats ( < 3 years old) in feral populations (16.8 vs 2.4%, P < 0.01), while the difference between the age groups was not statistically significant in domestic cats (2.4% vs 0.51%, P>0.05), all these results suggest that outdoor exposure time may be one of the most important factors for D. immitis prevalence in cats. Results reveal that D. immitis are prevalence in domestic and feral cats in northeastern China, which indicates that appropriate preventive measures should be taken to decrease the incidence of feline heartworm disease in Liaoning province, northeastern China.
Animals ; Cats* ; China* ; Dirofilaria immitis* ; Dirofilaria* ; Dirofilariasis ; Female ; Humans ; Incidence ; Male ; Prevalence ; Risk Factors ; Seroepidemiologic Studies*

Eukaryota ; Molecular Dynamics Simulation ; Permeability ; Protein Conformation ; Sodium ; metabolism ; Substrate Specificity ; Voltage-Gated Sodium Channels ; chemistry ; metabolism

Long noncoding RNAs (lncRNAs) are expressed in different species and different tissues, and perform different functions, but little is known about their involvement in the synthesis or secretion of follicle-stimulating hormone (FSH). In general, we have revealed lncRNA‍‒‍microRNA (miRNA)‍‒‍‍messenger RNA (mRNA) interactions that may play important roles in rat primary pituitary cells. In this study, a new lncRNA was identified for the first time. First, we analyzed the gene expression of lncRNA-m18as1 in different tissues and different stages by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and observed the localization of lncRNA-m18as1 with fluorescence in situ hybridization, which indicated that this lncRNA was distributed mainly in the cytoplasm. Next, we used RT-qPCR and enzyme-linked immunosorbent assay (ELISA) to analyze the regulation of FSH synthesis and secretion after overexpression or knockdown of lncRNA-m18as1 and found that lncRNA-m18as1 was positively correlated with FSH synthesis and secretion. In addition, mothers against decapentaplegic homolog 2 (Smad2) was highly expressed in our sequencing results. We also screened miR-18a-5p from our sequencing results as a miRNA that may bind to lncRNA-m18as1 and Smad2. We used RNA immunoprecipitation-qPCR (RIP-qPCR) and/or dual luciferase assays to confirm that lncRNA-m18as1 interacted with miR-18a-5p and miR-18a-5p interacted with Smad2. Fluorescence in situ hybridization (FISH) showed that lncRNA-m18as1 and miR-18a-5p were localized mainly in the cytoplasm. Finally, we determined the relationship among lncRNA-m18as1, miR-18a-5p, and the Smad2/3 pathway. Overall, we found that lncRNA-m18as1 acts as a molecular sponge of miR-18a-5p to regulate the synthesis and secretion of FSH through the Smad2/3 pathway.
Animals ; Cell Line, Tumor ; Cell Proliferation ; Follicle Stimulating Hormone/metabolism* ; Gene Expression Regulation, Neoplastic ; In Situ Hybridization, Fluorescence ; MicroRNAs/metabolism* ; RNA, Long Noncoding/metabolism* ; Rats

Astrocytes are the largest glial population in the mammalian brain. However, we have a minimal understanding of astrocyte development, especially fate specification in different regions of the brain. Through lineage tracing of the progenitors of the third ventricle (3V) wall via in-utero electroporation in the embryonic mouse brain, we show the fate specification and migration pattern of astrocytes derived from radial glia along the 3V wall. Unexpectedly, radial glia located in different regions along the 3V wall of the diencephalon produce distinct cell types: radial glia in the upper region produce astrocytes and those in the lower region produce neurons in the diencephalon. With genetic fate mapping analysis, we reveal that the first population of astrocytes appears along the zona incerta in the diencephalon. Astrogenesis occurs at an early time point in the dorsal region relative to that in the ventral region of the developing diencephalon. With transcriptomic analysis of the region-specific 3V wall and lateral ventricle (LV) wall, we identified cohorts of differentially-expressed genes in the dorsal 3V wall compared to the ventral 3V wall and LV wall that may regulate astrogenesis in the dorsal diencephalon. Together, these results demonstrate that the generation of astrocytes shows a spatiotemporal pattern in the developing mouse diencephalon.
Mice ; Animals ; Astrocytes ; Neuroglia/physiology* ; Diencephalon ; Brain ; Neurons ; Mammals