In order to explore the feasibility of inducing the human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) to differentiate into insulin-secreting cells with biological products alone, hUC-MSCs were separated and purified from the whole umbilical cord by the sequent digestion of collagenase II and trypsin followed by two-step centrifugation. hUC-MSCs were induced with IMDM culture medium containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and Ginkgo biloba extract (GBE). Before and after the induction, the morphological changes were observed under inverse microscope; the islet-related genes were detected by RT-PCR; islet-like clusters (ILCs) were identified by dithizone (DTZ) staining; PDX-1 and immunoreactive insulin (IRI) were examined by immunofluorescence method; the quantity and quality of IRI secretion were assayed by chemiluminescence immunoassay and Western blot respectively. The results showed that the purified hUC-MSCs presented long spindle-like shape and parallel or spiral arrangement which are typical morphological features of MSCs. After the induction, hUC-MSCs changed gradually into round or oval shape and gathered together to form ILCs; there were more than one hundred clusters on the growth surface of a flask of T25; ILCs were stained into positive mauve by DTZ and positive for PDX-1 and IRI; Western blot displayed that most of the IRI was proinsulin (PI). Therefore, hUC-MSCs can rapidly differentiate into insulin-secreting cells under the sole induction of EGF, bFGF, GBE and IMDM, but ILCs are not mature enough to produce sufficient true insulin.
Cell Differentiation ; Cells, Cultured ; Epidermal Growth Factor ; pharmacology ; Fibroblast Growth Factor 2 ; pharmacology ; Ginkgo biloba ; chemistry ; Humans ; Insulin-Secreting Cells ; cytology ; Mesenchymal Stromal Cells ; cytology ; Plant Extracts ; pharmacology ; Umbilical Cord ; cytology

Objective To investigate whether microRNA(miR)-199a-5p regulates blood-brain barrier(BBB)integrity through PI3K/Akt pathway after cerebral ischemia.Methods A permanent middle cerebral artery occlusion(MCAO)model was established in SPF adult male SD rats.Totally 48 rats were randomly divided into sham group(n=12),model group(n=12),MCAO+miR-199a-5p group(n=12),and MCAO+miR-199a-5p negative control group(n=12).The Ludmila Bellayev 12 point score was used to evaluate the neurobehavioral performance of rats;The integrity of the BBB after ischemia stroke was detected through Evans blue staining;Immunofluorescent staining was used to determine apoptosis after cerebral ischemia;Western blotting technology was used to detect the protein expression of claudin-5,phosphatidylinositol-3 kinase regulatory subunit 2(PIK3R2),p-Akt,Akt,and vascular endothelial growth factor(VEGF)-A;Real-time PCR was used to investigate the expression levels of miR-199a-5p,claudin-5,and VEGF-A in the ischemic penumbra and infarcted area of the brain.Results The result showed that miR-199a-5p mimic intervention improved proprioception and motor ability in MCAO rats.MiR-199a-5p mimic reduced the expression of PIK3R2 following ischemia stroke,activated the Akt signaling pathway,and increased the expression of claudin-5 and VEGF-A in the ischemic penumbra.In addition,miR-199a-5p alleviated inflammation after cerebral ischemia.MiR-199a-5p mimic reduced BBB permeability and reduced neuronal apoptosis after cerebral ischemia.Conclusion MiR-199a-5p can reduce the expression of PIK3R2 following ischemic stroke,activate the Akt signaling pathway,reduce the expression of inflammatory cytokines,and alleviate the damage to the blood-brain barrier.

BACKGROUND: Coronary atherosclerotic plaque could go through rapid progression and induce adverse cardiac events. This study aimed to evaluate the impacts of smoking status on clinical outcomes of coronary non-target lesions. METHODS: Consecutive patients with coronary heart disease who underwent two serial coronary angiographies were included. All coronary non-target lesions were recorded at first coronary angiography and analyzed using quantitative coronary angiography at both procedures. Patients were grouped into non-smokers, quitters, and smokers according to their smoking status. Clinical outcomes including rapid lesion progression, lesion re-vascularization, and myocardial infarction were recorded at second coronary angiography. Multivariable Cox regression analysis was used to investigate the association between smoking status and clinical outcomes. RESULTS: A total of 1255 patients and 1670 lesions were included. Smokers were younger and more likely to be male compared with non-smokers. Increase in percent diameter stenosis was significantly lower (2.7 [0.6, 7.1] % vs. 3.5 [0.9, 8.9]%) and 3.4 [1.1, 7.7]%, P = 0.020) in quitters than those in smokers and non-smokers. Quitters tended to have a decreased incidence of rapid lesions progression (15.8% [76/482] vs. 21.6% [74/342] and 20.6% [89/431], P = 0.062), lesion re-vascularization (13.1% [63/482] vs. 15.5% [53/432] and 15.5% [67/431], P = 0.448), lesion-related myocardial infarction (0.8% [4/482] vs. 2.6% [9/342] and 1.4% [6/431], P = 0.110) and all-cause myocardial infarction (1.9% [9/482] vs. 4.1% [14/342] and 2.3% [10/431], P = 0.128) compared with smokers and non-smokers. In multivariable analysis, smoking status was not an independent predictor for rapid lesion progression, lesion re-vascularization, and lesion-related myocardial infarction except that a higher risk of all-cause myocardial infarction was observed in smokers than non-smokers (hazards ratio: 3.00, 95% confidence interval: 1.04-8.62, P = 0.042). CONCLUSION Smoking cessation mitigates the increase in percent diameter stenosis of coronary non-target lesions, meanwhile, smokers are associated with increased risk for all-cause myocardial infarction compared with non-smokers.

Objective To compare left ventricular outflow tract (LVOT) gradient induced by dobutamine stress echocardiography (DSE) and exercise echocardiography (EE) in patients with hypertrophic cardiomyopathy (HCM).Methods DSE and EE were performed in 40 consecutive patients with HCM and LVOT gradient ＜50 mm Hg( 1mm Hg =0.133 kPa) at rest.Dobutamine was administered intravenously at incremental doses of 200,400,600,800 and 1200 p.g/min at 5 min intervals. LVOT gradients were measured at rest,at peak exercise and during each dose of DSE.Results LVOT gradient at rest was lower than 30 mm Hg in 36 patients and between 30 and 49 mm Hg in 4 patients.Of the 36 patients with LVOT gradient ＜ 30 mm Hg at rest,mechanical LVOT obstruction (latent LVOTO)was evidenced in 17 patients during EE and in 18 patients during DSE and good consistency (91.7％,kappa value 0.833) was found between the two maneuvers.The provoked gradient was similar between DSE at 800 μg/min and EE at peak exercise( P =0.181 ).In the 4 patients with LVOT gradient between 30 and 49 mm Hg[ (38.8 + 2.6)mm Hg],LVOT gradient increased to (85.3 +26.4) mm Hg during EE and (105.0 ±28.0) mm Hg during DES.Conclusions DES and EE are comparable and suitable provoke methods for identifying LVOT obstruction in patients with HCM.

Objectives: To study serum level of M2-muscarinic receptor autoantibody (M2-AAb) in hypertrophic cardiomyopathy (HCM) patients with its relationship to relevant clinical parameters. Methods: Our research included in 2 groups: HCM group, 133 patients and they were divided into 3 subgroups:Obstructive hypertrophic cardiomyopathy (HOCM) subgroup, 72, Latent obstructive hypertrophic cardiomyopathy (LHOCM) subgroup, 22 and Non-obstructive hypertrophic cardiomyopathy (NOCM) subgroup, 39; since there was no obstruction of left ventricular outflow tract (LVOT) in LHOCM and NOCM patients at resting, LHOCM and NOCM patients were combined as LHOCM+NOCM subgroup, 61 in comparison with HOCM subgroup. And Control group, 40 subjects had no organic heart disease and autoimmune diseases which were confirmed by 12 lead ECG, transthoracic echocardiography and routine hematological tests, they were not using β-blockers, glucocorticoids and immune-suppressants. Serum levels of M2-AAb were examined by ELISA, the relationship between M2-AAb and relevant clinical parameters were studied. Results: Compared with Control group, HCM group had increased serum level of M2-AAb [22.91 (17.21, 29.64) ng/ml] vs (17.14±5.66) ng/ml, P<0.01; M2-AAb was similar among HOCM, LHOCM and NOCM subgroups; M2-AAb in female patients were higher than male, P=0.001. Further investigation presented that the patients with family history of sudden death had the higher M2-AAb, P<0.05; patients with atrial fibrillation (AF) or left atrial diameter (LAD)≥50 mm or moderate to severe mitral regurgitation (MR) had the higher M2-AAb than those without such problems, all P<0.05. In HCM group, log M2-AAb was positively related to resting LVOT gradient (r=0.178, P=0.040); in HOCM subgroup, log M2-AAb was marginal positively related to resting LVOT gradient (r=0.224, P=0.058). Conclusions: Serum M2-AAb was elevated in HCM patients; gender, family history of sudden death may affect M2-AAb level; patients combining AF or LAD≥50 mm or moderate-severe MR had the higher M2-AAb and it was related to resting LVOT gradient.

Periplaneta americana is an important medicinal insect. A series of new drugs developed from it have remarkable clinical effects and are in great demand in the market. Because of unclear biology, the quality and yield of P. americana are affected. Understanding the developmental threshold temperature and effective accumulated temperature of P. americana can provide theoretical basis for standardized culture of P.americana. Under climate chamber, the threshold temperature and effective accumulated temperature for egg development of P. americana to were determined through effective accumulated temperature law. The threshold temperature was （15.8±0.71）°C, the effective accumulated temperature was 415.8±38.05 degree days. A model of the relationship between temperature and developmental rates was established.
Animals ; Ovum ; physiology ; Periplaneta ; physiology ; Temperature

Clear cell renal cell carcinoma (ccRCC) has been proved to be a metabolic disease with high