Objective To compare four different methods for extracting exosomes derived from human umbilical cord mesenchymal stem cells(HUCMSCs-Exos) and obtain a simple, efficient, and cost-effective method for isolating exosomes.Methods The(Human umbilical cord Mesenchymal stem cells,HUC-MSCs)were subcultured, observed for cell morphology under microscope, and detected for cell proliferation ability, specific surface antigen expression level and multidirectional differentiation potential. The hucMSCs-Exos were extracted from the culture supernatant of HUC-MSCs by four methods, ultracentrifugation, polyethylene glycol(PEG) precipitation, sucrose precipitation and Total Exosome Isolation kit. The exosomes were observed for the morphology by transmission electron microscope, measured for the concentration and particle size by nanoparticle tracking analysis(NTA), detected for the total protein concentration by BCA, and detected for the specific marker protein level by Western blot.Results The subcultured HUC-MSCs showed long spindle shape and vortex adherence growth under microscope with fast cell proliferation, highly expressed MSCs specific surface antigens CD73 and CD90,and extremely low expressed hematopoietic stem cell surface markers CD34 and CD45, which had the potential to differentiate into adipocytes and osteoblasts. The HUCMSCs-Exos were extracted by all the four methods, except kit method, the exosomes obtained by the other three methods all displayed a cup-shaped double-layer membrane structure with clear boundaries. The total protein concentration and particle concentration of exosomes isolated using the kit were significantly higher than those of the other three methods(F = 35. 18, each P < 0. 01). The exosomes extracted by the four methods all expressed specific marker proteins CD63 and CD81. Except the exosomes extracted by sucrose precipitation did not express CD9, the exosomes extracted by the other methods all expressed CD9.Conclusion The exosomes can be extracted by all the four methods, and the quality of exosomes extracted by ultracentrifugation is stable; the yield of exosomes extracted by the kit is higher, more time-saving and more convenient; the purity of exosomes extracted by PEG precipitation and sucrose precipitation is higher. According to different production requirements, the appropriate exosomes isolation techniques can be selected.

Objective To study the application of the two-paths double U-shaped fixer in clinical intravenous indwelling needle fixation. Methods A total of 420 cases using of intravenous indwelling needle from January to June 2015 were involved and divided into experimental group(210 cases) and control group(210 cases) by random digits table method. The two-paths double U-shaped fixer was applied in experimental group and the control group were used tape by Y type to fix. The incidence of returning-blood, blocking, phlebitis, slippage and the comfort between two groups were observed and compared. Results The incidence of returning-blood, blocking, phlebitis, slippage in experimental group were 6.2%(13/210), 0.5%(1/210), 0 , which were lower than those in control group 48.6%(102/210), 5.7%(12/210), 1.4%(3/210) seperately, the differences were statistically significant (χ2=117.895, P<0.01). The incidence of phlebitis levelⅠ,Ⅱ,ⅢandⅣin experimental group were 3.3%(7/210), 1.4%(3/210), 0.9%(2/210), 0, which were lower than those in control group 11.9%(25/210), 9.0%(19/210), 3.8%(8/210), 2.4%(5/210), the differences were statistically significant(Z=-5.960, P<0.01). The incidence of the comfort level 1, 2, 3 and 4 in experimental group were 92.4%(194/210), 6.2%(13/210), 1.4%(3/210), 0 individually, which were higher than those in control group 36.7%(77/210), 46.7%(98/210), 12.4%(26/210), 4.3%(9/210), the differences were statistical significant (Z=-16.228, P < 0.01). Conclusions Two-paths double U-shaped fixationis is better than Y-shaped tape fixation. To compare the venous indwelling needle fixation, two-paths double U-shaped fixer could fix more effectively and safety. At the same time, it also has advantages in convenient, observation and appearance. It could be widely used in clinical practice.

Parkinson's disease(PD) is a progressive neurodegenerative disorder harming mainly the elderly.Astrocyte may be involved in the pathogenesis of PD.However,it also plays a protective role in PD by releasing neurotrophic factors and glutathione to support the survival of dopaminergic neurons.Astrocyte may also release proinflammatory cytokines to promote the development of PD.

Objective:To investigate the electromyography activities of the facial and neck muscles during mandibular movement in children with high-angle.Methods:A computer-integrated BioEMG Ⅱ diagnostic system was used to synchronously record the electromyography activities of temporal anterior(TA), masseter muscle(MM), sternocleidomastoid(SCM) and digastric anterior(DA) muscles during mandibular movement in 14 children with high-angle and 16 children with normal-angle.Results:Muscular activity of TA, DA and SCM in children with high-angle was significantly weaker than that in those with normal-angle in the rest position(P

After intracerebral ventricular administration of adrenomedullin (AM) in normal saline (NS) or 30% glucose to rats, the effects on blood glucose and glucose regulatory hormones were studied and compared. 1) Am in NS increased significantly the concentration of plasma insulin and C peptide, and decreased the concentration of glucagon and blood glucose. 2) As compared with 30% glucose alone, AM in 30% glucose raised significantly the plasma insulin and C peptide and decreased significantly the glucagon concentration, but the blood glucose level remained unchanged. The results showed that the AM plays a role in the regulation of pancreatic hormone secretion by central nerve system.

This article analyzes the forward and backward feedback pathways between cones and horizontal cells in the Cascade model and sets up a Hopfield Network to simulate the interaction mechanism.Additionally,it appears that the phase character of horizontal cell is connected with spectrum and a CMAC Network is established to simulate the mapping process.This article aims to indicate that the mapping of horizontal cell and spectrum might be the underlying way for retina to code the signal from spectral input.

Objective To determine the prevalence of renal artery stenosis (RAS) and the efficacy of percutaneous transluminal renal angioplasty (PTRA) and stenting in patients with documented coronary artery disease.Methods Selective renal artery angiography or abdominal digital subtraction angiography was performed immediately after coronary angiography to detect and quantify RAS in 200 consecutive patients with known or suspected coronary artery disease. Eight severe stenotic renal arteries in 7 patients were treated with PTRA and stenting. All the lesions are considered to be atherosclerotic. Results RAS (defined as ≥50% diameter stenosis) was detected in 21 of 200 patients (10.5%) and 19 of 99 patients with coronary artery disease (19.2%) who had at least one coronary artery of ≥50% diameter stenosis. Procedure success rate was 100% in the 7 patients received PTRA and stenting. Blood pressure control improved in 74.1% patients with hypertension. Angiographic follow up was done in 3 patients in 3 to 9 months and no restenosis was found in the vessels.Conclusions It is necessary to routinely perform RAS for the patients with coronary artery discase undergoing coronary angiography to identify RAS, especially in those associated with hypertension, so as to diagnose and treat RAS in early stage. PTRA and stenting carries high success rate and appears to be an effective and safe treatment for RAS.

After intracerebral ventricular administration of GLP-1 in NS or 30% glucose to rats,their effects on blood glucose and glucose regulatory hormones were observed. The results demonstrated:1. Intracerebral ventricular administration of GLP-1 in NS didn't influence plasma stucose, insulin and glucagon secretion.2. Intracerebral ventricular administration of GLP-1 in 30% glucose increased plasma glucose concentration significantly and decreased insulin concentration,but no change of glucagon concentration. This experiment showed that GLP-1 might participate in CNS regulation of pancreatic islet hormonesecretion.

AIM:To observe the influence of beta-amyloid precursor protein (APP17) on the study ability, memory and the expression of neurotrophin-3 (NT-3), nerve growth factor(NGF)in the hippocampus neuron of the model mice. METHODS: Mice brain aging model were produced with D-galactose(D-gal), the model mice were given hypodermic injection of APP17 peptide. APP17 peptide is the 319-335 peptide sequence of beta-amyloid precursor protein. Eight weeks later, the animals were observed by water labyrinth test and immunohistochemistry assay. RESULT:(1) The whole time needed and total times of wrong response for the D-gal group mice to complete the whole course of the water labyrinth test is significantly higher than the normal control group. (2) The expression of NT-3, NGF in the hippocampus neurons of the mice in APP17 peptide group is significantly higher than that of the normal control group and D-gal mice group, P

Objective To investigate whether there is any functional link between p27~(Kip1) function and all-trans retinoic acid (RA) in the control of neuronal differentiation of immortalized human neural progenitor cells (hSN12W-TERT cells). To investigate the mechanism by which p27~(Kip1) regulates the differentiation of immortalized human neural progenitor cells. Methods hSN12W-TERT cells were derived from the striatums of human embryos at 12 weeks gestation and cultured with serum-free medium in presence of EGF and bFGF. At the appropriated time, hSN12W-TERT cells were exposed to 1μmol/L RA for 3, 5, 7 days respectively. The experiment was repeated there times. Cell cycle analysis was performed by flow cytometry analysis (FACS). The expression of p27~(Kip1), p21~(cip1), cyclin-dependent kinase 2 (cdk2), p-cdk2 and S-phase kinase-associated protein 2 (skp2) in hSN12W-TERT cells before and after RA treatment cells were determined by using Western blotting analysis. Results FACS result showed that 77.25% of proliferating hSN12W-TERT cells were in the G1/G0-phase while 9.38% of cells in the S-phase. Following RA treatment, cell growth was arrested, and 85.68% of cells accumulated in G1/G0-phase while 8.57% of cells in the S-phase. Western blotting analysis demonstrated that the levels of p27~(Kip1) in the hSN12W-TERT cells increased following 3 days' treatment with RA compared with those of normal untreated cells, with a peak at 5 days (P<0.05). The similar results were acquired both in nuclear proteins and in cytoplasm proteins of hSN12W-TERT cells. The expression level of p21~(cip1) decreased in response to RA treatment. RA did not affect the expression of cdk2, but the expression of p-cdk2, which represented the activity of cdk2, was markedly decreased in response to RA treatment. Skp2, which was required for the ubiquitin-mediated degradation of p27~(Kip1), was detected in proliferating hSN12W-TERT cells. The expression of skp2 reduced dramatically in response to RA treatment in a time-dependent manner.Conclusion There is a functional link between RA and p27~(Kip1) function in the control of neuronal differentiation in hSN12W-TERT cells. P27~(Kip1) plays a key role during neuronal differentiation. Moreover, high levels of p27~(Kip1) are associated with its degradation inhibiting through reducing proteasome-dependent proteolysis.