Objective To study the vascular endothelial growth factor(VEGF) expression and secretion of mesenchymal stem cells(MSCs) of rabbit transfected with pcDNA3-VEGF165 expression plasmid in order to construct a kind of tissue engineering artificial skull with more blood supply.Methods By gene reconstruction method,the VEGF165 gene was cloned into eukaryotic expression plasmid pcDNA3 and recombined eukaryotic expression plasmid pcDNA3-VEGF165 was constructed;By lipofectamine transfection method,pcDNA3-VEGF165 expression plasmid was transfected into MSCs of rabbit;By RT-PCR and Western blotting methods,the VEGF mRNA expression and VEGF protein secretion in the MSCs were detected.Results Recombined eukaryotic expression plasmid pcDNA3-VEGF165 was confirmed to be true by double enzyme digestion,two strips came to appear in 5400 bp and 600 bp of gelose electrophoresis and their sizes accorded with pcDNA3 plasmid and VEGF165 accordingly;Primarily cultured and subcultured MSCs of rabbit were successfully performed and the MSCs storehouse of rabbit was established,primary MSCs presented lymphoid form at first and then the morphology of them became circulara,polygonal or irregular forms,they were more and more like fibroblastic cells after subcultured cultivation.The expressions of VEGF mRNA and VEGF protein in the MSCs were found after transiently transfection by RT-PCR and Western blotting methods.Conclusion Recombined eukaryotic expression plasmid pcDNA3VEGF165 can be transfected into MSCs of rabbit effectively by lipofectamine and the VEGF expression can be detected in the MSCs after transfection.

Humans ; Interferons ; administration & dosage ; therapeutic use ; Lymphoma, Mantle-Cell ; drug therapy ; Male ; Middle Aged ; Thalidomide ; administration & dosage ; therapeutic use

A total of 183 patients of ectopic pregnancy due to tubal factors were divided into the methotrexate (MTX),conservative surgery and salpingectomy groups.The dose of gonadotropin,counts of harvested oocytes and high-quality embryos and pregnancy rate were compared among three groups to analyze the differences of in vitro fertilization and embryo transfer (IVF-ET).And the above parameters showed no significant differences (P ＞0.05 ).The clinical pregnancy rate of the conservative group was higher than the other two groups.And the difference was statistically significant compared with salpingectomy ( P ＜ 0.05 ).It suggested that the treatments of ectopic pregnancy had some effects on the outcome of IVF-ET.The pregnancy rate was slightly higher in the conservative surgery group.

Objective To investigate the impact of ultrapure dialysate on long-term survival rate and complication in maintenance hemodialysis patients.Methods Seventy maintenance hemodialysis patients were selected,and they were divided into ultrapure dialysate group (36 cases) and control group (34 cases) by random digits table method.The 1-,3-,5-year mortality,death causes and the incidences of cardiovascular complication,infection,cerebrovascular accident were recorded.Results The 3-and 5-year mortality in ultrapure dialysate group were significantly lower than those in control group:30.6％ (11/36) vs.41.2％ (14/34) and 41.7％ (15/36) vs.82.4％ (28/34),and there were statistical differences (P ＜ 0.05).The mortality of infection in ultrapure dialysate group was significantly lower than that in control group:1/15 vs.21.4％ (6/28),and there was statistical difference (P ＜ 0.05).The incidences of cardiovascular complication,cerebrovascular accident,infection in ultrapure dialysate group were significantly lower than those in control group:47.2％ (17/36) vs.79.4％ (27/34),27.8％ (10/36) vs.41.2％ (14/34) and 41.7％ (15/36) vs.73.5％ (25/34),and there were statistical differences (P ＜ 0.05).Conclusion Ultrapure dialysate can increase long-term survival rate and decrease incidence of complication in maintenance hemodialysis patients.

Objective To observe the effect of electro-acupuncture therapy (ET) on the expression of sodium channel Na(v) 1.1 in rats after acute cerebral ischemia and the mechanism of any protective function of ET.Methods A model of focal acute cerebral ischemia was established by occluding the right middle cerebral artery.One hundred and eighty healthy SD rats were randomly divided into a sham operation control (SC) group, an ischemia control (IC) group, a real ET group and a false ET group, with 45 in each group. Immunohistochemistry and real-time polymerase chain reaction (PGR) methods were used to detect Na(v)1. 1 expression. 2,3,5-triphenyl tetrazolium chloride (TTC) staining was used to detect infarct volume. Neurological examination and grading was carried out at 6 hours and then 1, 2, 3 and 7 days after inducing ischemia. Results The gradings and infarction volume ratios of the rats in the IC group were the most serious, while in the real ET group the severity was much less at each time point. Compared with the SC group, the expression of Na(v) 1.1 was significantly up-regulated in the IC group. The expression of Na(v) 1.1 was increased at the 6th hour, then down-regulated to the lowest level at day 1,then from the 2nd to the 7th day was up-regulated again. The expression of Na(v) 1.1 in the real ET group was significantly lower than in the IC group. Although the expression of Na(v)1.1 in the false ET group was low compared with the IC group, the difference was not significant. The difference between the real ET group and the false ET group was significant, however. Conclusions ET can reduce damage from cerebral ischemia and benefit the recovery of neural function. ET can also could regulate the expression of Na(v)1.1 after acute cerebral ischemia, which may be an important mechanism for neural function recovery.

Objective To investigate the correlation between plasma homocysteine (Hcy) level and cerebral microbleeds (CMBs) band leukoaraiosis (LA) in patients with acute stroke.Methods The clinical and imaging data of patients with acute stroke were analyzed retrospectively.The numbers of CMBs were counted and the severity of LA was graded according to the results of MRI.Fasting venous samples were obtained and the plasma Hey concentration was measured the next day after admission.Results A total of 139 patients with acute stroke were enrolled,67 of them were females and 72 were males (mean age 70.1 ± 10.2 years); 24 had hemorrhagic stroke and 115 had ischemic stroke.The age (76.23 ± 8.74 years vs.64.58 ± 7.42 years;t =4.621,P =0.012) hypertension ratio (89.13％ vs.67.74％ ;x2 =8.324,P =0.0 370) and plasma Hey level (14.53 ± 4.31 mmol/L vs.11.31 ±3.16 mmol/L;t =6.538,P=0.008| in a severe LA group (n=46) were significantly higher than those in a non-severe LA group (n =93).Spearman correlation analysis showed that there was significant correlation between the plasma Hcy level and the severity of LA (rs =0.365,P =0.002).Multivariate logistic regression analysis showed that the increased Hey level (odds ratio [ OR ],1.366,95％confidence interval [ CI] 1.141 - 1.526; P =0.010) and age (OR 1.093,95％ CI 1.031 - 1.162; P =0.016)were the independent risk factors for severe LA.The age (74.37 ± 6.35 years vs.67.56 ± 8.52 years; t =6.628,P =0.038) and hypertension ratio (94.74％ vs.62.20％;x2 =8.773,P =0.002) in a CBM group were significantly higher than those in a non-CMB group (n =82).Spearman correlation analysis showed that there was no significant correlation between the plasma Hcy level and the numbers of CBMs (rs =0.038,P =0.813).Multivariate logistic regression analysis showed that hypertension was an independent risk factor for CBMs.Conclusions The elevated plasma Hcy level was associated with LA,but it was not associated with CBMs.

Objective:To isolate cell spheres containing cancer stem cells (CSCs) from lung cancer cell lines (A549 and H1299) and iden-tify their biological characteristics. Methods:By adopting the method of serum-free suspension culture, A549 cells and H1299 cells were cultured on no-adhesion plate to form tumor spheres. Clone assay, CCK-8 assay, and Transwell assay were employed to observe proliferation, self-renewal and invasion of tumor spheres. Besides, RT-PCR was performed to compare the expression levels of stem cell markers between sphere cells and adherent cells. Adherent A549 cells and A549 cell spheres were inoculated subcutaneously in nude mice and the tumor growth was assessed. Results:Isolated CSCs from A549 cells and H1299 cells in serum-free medium (SFM) without adhesion could grow as floating cell spheres. The results demonstrated that the self-renewal, proliferation and invasion of A549 and H1299 sphere cells were stronger than parent cells (P<0.05). When compared with adherent cells, the mRNA level of expres-sion of cell spheres' stem cell markers (Oct4 and Nanog) were significantly high (P<0.05) and A549 spheres had a stronger tumorigenici-ty in nude mice. Conclusion:SFM without adhesion can be a quick and easy method to construct stem cells model from human lung adenocarcinoma cell lines A549 and H1299.

Caliciviridae Infections ; epidemiology ; Child, Hospitalized ; Child, Preschool ; China ; epidemiology ; Female ; Gastroenteritis ; epidemiology ; virology ; Humans ; Infant ; Male ; Sapovirus ; isolation & purification

Adenoviridae ; classification ; genetics ; pathogenicity ; Adenoviridae Infections ; epidemiology ; prevention & control ; virology ; Age Distribution ; Child, Preschool ; Humans ; Infant ; Infant, Newborn

Objective To investigate the probability of verifying the X-knife treatment plan by the radiotherapy simulator,and to analyze and report the errors of the X-knife treatment plan for reference.Methods The radiation fields of the X-knife treatment plan were observed in the whole simulating treatment process by the radiotherapy simulator.Results The error rate of X-knife treatment plan were in the simulating treatment process was 5.9%(22/372).Among the errors,3 cases of X-knife plans were found in the simulating treatment process,and the errors of isocenter in Z direction were 10cm.Conclusion This is a special error of X-knife TPS,which deserves more attention in stereotactic radiotherapy.