Objective To investigate the differentiation from human mesenchymal stem cells (hMSC) into neuron-like cells. Methods hMSC were separated from rib marrow with Ficoll-Paque reagent and expanded in culture medium. hMSC were induced to differentiate into neurons with DMEM/BHA/DMSO or DMEM/monothioglycerol, respectively. Neuron-specific enolase (NSE), neurofilament (NF), nestin, glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. Results hMSC were expanded to be undifferentiated cells in culture for more than 10 passages. The isolated and cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. Simple method induced hMSC exhibiting a neuronal phenotype, with a positive expression of NSE, NF-M and nestin at 5 hours. But the neuron-like cells did not express the glial astrocyte marker GFAP. Conclusion It suggests that hMSC can be differentiated into neurons in vitro .

OBJECTIVE:To understand the distribution and drug resistance of the pathogens by sputum culture in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) in our hospital so as to provide reference for rational use of antibiotics. METHODS:From Dec. 2010 to Dec. 2014,the sputum specimens were collected from the AECOPD patients,then the identification of 307 strains of pathogens and drug susceptibility test were carried out,and the data were analyzed statistically by using SPSS 17.0 software. RESULTS:A total of 307 strains of pathogens were collected,of which 17 cases of gram-positive ba-cillus accounted (5.54%),247 cases of gram negative cocci (80.46%),43 cases of fungi accounted (14.00%). The most com-mon isolates from sputum specimens were Pseudomonas aeruginosa (33.22%),Acinetobacter baumannii (19.54%),Stenotroph-omonas maltophilia(9.77%),Klebsiella pneumoniae(7.82%),Candida albicans(6.84%),etc. P. aeruginosa and A. baumannii were highly multidrug-resistant. There were 10 strains of ESBLs-producing K. pneumonia isolated, with the isolation rate of 41.67%. No Staphylococcus aureus strain was found resistant to vancomycin,teicoplanin or linezolid. Methicillin resistant strains in S. aureus(MRSA)accounted for 50.00%. CONCLUSIONS:Gram-negative bacilli are the most common pathogens in the AE-COPD patients. The common species of pathogens are highly resistant. More attention should be paid to the drug resistance monitor-ing of pathogens and rational use of antibiotics according to the results of susceptibility test.

Objective: To study the interaction between the calcium channel antagonist nimodipine and growth factors (vascular endothelial growth factor [VEGF] and platelet-derived growth factor[PDGF]) during the formation of new retinal vessels. Methods: The hyperoxia model was induced by proliferative retinopathy (OIR) in newborn Sprague-Dawley (SD) rats. SD rats (2 d after birth) were randomized into 5 groups: normal control group, pure OIR group and group 3, 4 and 5, where the animals received retrobulbar injection of nimodipine 10 ?l, 5 ?l and 2 ?l once every 2 d for 3 times, respectively. Both eyeballs of newborn rats were made into common pathological sections and detected by immunohistochemistry method to count the nuclei of proliferative retinal vessel cells and to investigate the expression of VEGF and PDGF in retina. Results: The nuclei of proliferative retinal vessel cells and the expressions of VEGF and PDGF in pure OIR group increased significantly compared to those of normal control group(P

AIM: To investigate the differentiation from human mesenchymal stem cells (hMSC) to adipocytes.METHODS: hMSC were separated from rib marrow and expanded in culture medium. To detect the surface antigens, the labeled cells were analysed on a FACScan flow cytometer. hMSC were induced with dexamethasone, insulin, 1-methy1-3-isobutylxanthine and indomethacin which acted as adipocyte differentiation inducer. The cells were stained with Oil Red O. The number of adipocytes were counted on a phase-contrast microscope.RESULTS: hMSC were expanded as undifferentiated cells in culture for more than 5 passages. The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. These expanded, attached MSC were uniformly positive for CD29, CD44, CD90, CD105, CD166 and didn't express CD14, CD34, CD45, CD11a. After induced with induction medium, lipid vacuoles were first detectable within the cells at 48 hours. Two weeks later, more than 85% MSC differentiated into adipocytes which displayed a perinuclear accumlation of lipid vacuoles, as detected by Oil Red O. CONCLUSION: hMSC can be induced to differentiate into adipocytes. [

AIM: To investigate the differentiation from human mesenchymal stem cells (hMSC) into osteoblasts. METHODS: MSC were separated from human marrow with Ficoll-Paque reagent and expanded in cuture medium. To detect the surface antigens, The labeled cells were analysed on a FACScan flow cytometer. hMSC were induced to differentiate from mesenchymal stem cells into osteoblasts with dexamethasone, vitamin C, ?-GP. Cell morphology?AP activity?calcium deposition and osteopontin were detected. P10 MSC were compared to P3 MSC in the tendency of osteoblastic differentiation. RESULTS: The cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. hMSC showed a strong self-renewal capacity. After primary culture, approximately (5-6)?10 5 cells were obtained. These expanded attached MSC were uniformaly positive for CD29,CD44,CD59,CD105,CD166 and didn't express CD11a, CD14, CD33, CD34, CD45, CD38, CD80, CD86, CD117. After osteoblasts induction, the cells changed from spindle-shape to cuboidal and polygonal in cell morphology. The AP activity increased gradually and many scattered calcium nodes were observed. The expression of osteopontin was positive. CONCLUSION: hMSC can be induced to differentiate into osteoblasts.

AIM: To investigate the effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts.METHODS: hMSC were separated from human marrow and expanded in cuture medium. hMSC were induced with dexamethasone, ?-glycerophosphate, vitamin C which acted as osteoblast differentiation inducer. PD98059 was added into the osteoblasts induction medium. The cells were assayed with cell morphology, alkaline phosphatase (AP) activity and calcium deposition. RESULTS: The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. After induced with osteoblasts induction medium, the cells showed changes in cell morphology from spindle-shape to cuboidal and polygonal. The AP activity increased gradually and reached the peak in 12 days, then decreased. Many scattered tangerice calcium nodes were observed. PD 98059 significantly inhibited AP activity and calcium deposition in a dose-dependent manner. A striking observation of the present study was that a few adipocytes appeared in cultures that were treated with PD 98059 and osteogenic differentiation medium. CONCLUSION: These results indicated that osteogenic diferentiation from the hMSCs was related to the activation of the ERK.inCwhichactedasost

AIM: To investigate the differentiation from human mesenchymal stem cells(hMSC) into neuron-like cells by the increase in intracellular cyclic AMP. METHODS: hMSC were separated from human marrow with Ficoll-Paque reagent and expanded in culture medium. hMSC were induced to differentiate into neurons with Forskolin and 3-isobutyl-1-methyl-xanthine (IBMX). Neuron-specific enolase(NSE), neurofilament(NF), glial fibrillary acaidic protein(GFAP) were detected by immunohistochemistry. RESULTS: hMSC were expanded as undifferentiated cells in culture for more than 10 passages. Forskolin/IBMX can induce hMSC to exhibit a neuronal phenotype, expressing NSE and NF-M in 5 hours. But the neuron-like cells didn't express the glial astrocyte marker GFAP. CONCLUSION: hMSC can be induced to differentiate into neurons by increase in the intracellular cAMP.

Objective To observe the value of contrast-enhanced ultrasonography (CEUS) in quantitatively evaluating the evolution of the blood supply of rabbit liver VX2 tumor models in early stage. Methods Eighteen liver VX2 tumor models of rabbits were divided into 3 groups randomly (each n=6). CEUS was performed on the 10th, 15th and 20th day group after establishemnt, resepectively. Time intensity curve (TIC) was plotted, the rabbits' liver tumor and surrounding normal liver tissue were resected for immuno-histochemistry and microvessel density (MVD) count. Results CEUS showed that the rabbit liver VX2 tumor present a typical malignant tumor imaging fast-in and fast-out in early stage. TIC showed that the time to peak (TTP) of hepatic tumor is shorter than that of surrounding normal liver tissue in all three groups (P＜0.01), no statistical difference of tumor's TTP was detected among three groups (P>0.05). Absolute enhanced intensity (AEI) and MVD reduced in turn of 10th, 15th and 20th day group. MVD of tumor was higher in 10thday group than surrounding normal liver tissue (P＜0.01), while was equal in 15th day group (P>0.05) and lower in 20th day group (P＜0.01). AEI of liver tumor had positive correlation with MVD (r=0.83, P＜0.05). Conclusion AEI of TIC can be used to quantitatively assess the evolution of the blood supply in rabbit liver VX2 tumor models. The blood supply in early stage of rabbit liver VX2 tumor models shows a gradual descent process from abundant blood supply to medium blood supply and then low blood supply.

AIM:To investigate the differentiation from human mesenchymal stem cells(hMSC) into neuron-like cells.METHODS:hMSC were separated from rib marrow with Ficoll-Paque reagent and expanded in culture medium. hMSC were induced to differentiate with DMEM/monothioglycerol or DMEM/β-mercaptoethanol, respectively. Neuron-specific enolase(NSE), neurofilament(NF), and glial fibrillary acidic protein(GFAP) were detected by immunohistochemistry. RESULTS:hMSC were expanded as undifferentiated cells in culture for more than 5 passages. When treated with monothioglycerol or β-mercaptoethanol for 5 hours, hMSC exhibited neuronal phenotype. The expression of NSE and NF in the neuron-like cells was positive, but the glial astrocyte marker GFAP didn't express. CONCLUSION:hMSC can be induced to differentiate into neurous.

Objective To observe the application of the mini-mental status examination (MMSE)cognitive assessment in the patients with brain glioma before and after surgery. Methods Using MMSE,36 pastients with primary brain glioma were subjected to the cognitive assessment before surgery, after surgery,and 3 monthsr after surgery. Results The quantitative cognitive assessments with MMSE before surgery revealed the hidden cognitive dysfunction patients.The quantitative cognitive assessments after surgery showed that surgeons might need to protect the non-function area and to form the idea of cognitive function in patients with glioma.Conclusion MMSE assessment is a simple,understandably,and convenient method having good compliance of patient. It may be effectively used to assess cognitive impairment for patients with glioma and worth being studied continuously and used widely in the clinic practice.