Objective To observe the effect of subretinal injection of retinal pigment epithelium (RPE) cells for RPE in mice.Methods A total of 30 postnatal day 7 C57BL/6J mice were randomly divided into normal mice group,OIR model group and OIR model cell transplanted group,10 mice in each group.The OIR model was induced in mice of OIR model group and OIR model cell transplanted group.The RPE cells were subretinal injected into the RPE of mice in OIR model cell transplanted group.At 20 days after the injection,the RPE thickness was evaluated by fluorescence microscope.The expression of RPE65,Bestrophin and zonula occludens-1 (ZO-1) were estimated by Western blot and real-time quantitative PCR (RT-PCR).Results The thickness of RPE in OIR model mice was thinner than that in normal mice;the thickness of RPE in OIR model cell transplantation mice was significantly thicker than that in the OIR model mice.The results of Western blot and RT-PCR indicated that the differences of protein (F=8.597,18.864,25.691) and mRNA expression (F=39.458,11.461,34.796) of RPE65,Bestrophin,ZO-1 were statistically significant between OIR model group and OIR model cell transplanted group (P＜0.05).Conclusions Subretinal injection of RPE cells can promote RPE thickening.RPE65 and Bestrophin protein relative expression levels increased,ZO-1 protein relative expression levels reduced;mRNA expression levels of RPE65,Bestrophin and ZO-1 genes increased.

AIM: To investigate the effects of microRNA (miRNA)-483-3p on the growth and migration of human glioma cell line A172 and its potential mechanisms.METHODS: The abundance of miRNA-483-3p in human embryonic kidney 293 cells and different human glioma cell lines (A172, U251 and SHG44) was measured by RT-qPCR.After down-regulation of miRNA-483-3p by transfection of inhibitor in the A172 cells, the cell viability, cell cycle distribution and cell migration were detected by CCK-8 assay, flow cytometry and Transwell assay, respectively.Furthermore, the protein levels of cell cycle-related molecules and epithelial-mesenchymal transition markers were measured by Western blot.Luciferase reporter assay was used to predict and verify the target gene of miRNA-483-3p.RESULTS: miRNA-483-3p was highly expressed in human glioma cells.Knockdown of miRNA-483-3p inhibited A172 cell viability, arrested cell cycle and decreased cell migration rate.Furthermore, the protein levels of cyclin D1, cyclin-dependent kinase 4, phoshorylated retinoblastoma protein, N-cadherin and vimentin were significantly decreased after knockdown of miRNA-483-3p, accompanied with the up-regulation of E-cadherin and β-catenin protein expression.Luciferase reporter assay demonstrated that Smad4 was a potential target gene of miRNA-483-3p.Down-regulation of Smad4 in the A172 cells transfected with miRNA-483-3p inhibitor partially reversed the effect of miRNA-483-3p on cell viability and migration.CONCLUSION: Knockdown of miRNA-483-3p restrains the growth and migration of A172 cells by targeting Smad4.

Burns ; complications ; immunology ; physiopathology ; Humans ; Hypoxia ; etiology ; immunology ; physiopathology ; Ischemia ; etiology ; immunology ; physiopathology

Chronic Disease ; therapy ; Constipation ; diagnosis ; therapy ; Gastrointestinal Transit ; Humans

According to the synonymous codons used in 28 open reading frames from Pichia pastoris, the codon usage in this species was calculated and 19 codons have been inferred to be its optimal codons. The results show that pattern of the codon usage in P. pastoris is similar to that in S. cerevisiae and in K. lactis except for the synonymous codon of glutamic acid, which may be the special bias of P. pastoris.

Acute Coronary Syndrome ; therapy ; Angioplasty, Balloon, Coronary ; Humans ; No-Reflow Phenomenon ; prevention & control

Objective:To investigate the efficacy of postoperative chemotherapy in patients with early-stage pulmonary large cell neu-roendocrine carcinoma (LCNEC) after resection. Methods:A cohort of 50 patients who underwent resection and systematic nodal dissection for LCNEC between January 2008 and December 2014 in our institution was retrospectively reviewed. The patients were divided into adjuvant chemotherapy group (32 cases) and non-chemotherapy group (18 cases). Follow-up information was investigated. Results:The median survival and the 5-year survival rate were 48 months and 72.5%for the adjuvant chemotherapy group whereas 29 months and 35.6%for the non-adjuvant chemotherapy group, respectively. Univariate and multivariate analyses using Cox's proportional hazard models showed that postoperative chemotherapy was a signifi cant prognostic factor for OS (P=0.005;hazard ratio=0.281, P=0.008, respectively). Conclusion:Postoperative chemotherapy is beneficial to patients with early-stage pulmonary LCNEC after complete resection.

[Objective]To evaluate the effect of continuous dynamic pressure stress on experimental humeral fracture healing and to explore the role of COX-2 /PGE2 /cAMP signaling pathway in healing promotion induced by the special biomechanical condition.[Method]A total of 120 New Zealand rabbits were divided randomly into three groups: Group A,unilateral humeral fracture was fixed with shape memory connector(SMC),producing axial continuous dynamic pressure stress on fracture gap.Group B,fixed with 4-hole DCP,and Group C,fixed with SMC and gavaged with aqueous suspensions of celecoxib(3 mg/kg/day) after fracture.COX-1 and COX-2 mRNA expressions in fracture gap were determined by real-time RT-PCR at 0,3,7,14,21,28 and 56 days after operation,as well as the contents of PGE2 and cAMP were measured by radioimmunoassay(RIA).Bone-specific alkaline phosphatase(B-ALP) activity was determined by measuring p-nitrophenol and osteocalcin concentrations were measured using enzyme-linked immunosorbent assay(ELISA) for serial collected blood samples.Callus specimens fixed by formalin were used for histopathological examination.[Result]The COX-2 mRNA levels normalized with GAPDH mRNA as well as the contents of PGE2 and cAMP showed significant difference along with the 56-day period and also between the two internal fixations.Histopathological observation and determination of serum biochemical markers of bone formation revealed that the endochondral bone formation and the callus remodelling took place earlier in Group A than in Group B.The accelerated healing process and increased serum values of the bone formation markers induced by continuous dynamic pressure stress of SMC were inhibited by celecoxib,a specific COX-2 inhibitor.[Conclusion]The persistent dynamic longitudinal pressure stress produced by SMC contributed to endochondral bone formation and callus remodelling,resulting in an acceleration of fracture healing,which was considered to be closely correlated with the COX-2/PGE2/cAMP signal transduction pathway.

Objective To investigate the histomorphology,cellular biocompatibility in vitro and tensile force resistance property of fibrin-binding amniotic membrane(FBAM)and to explore its feasibility and superiority using as ocular surface graft.Methods After observing the morphology of FBAM under light microscope and scanning electron microscope,we did primary culture of corneal epithelial cells,taking monolayer fibrin-binding amniotic membrane as carrier,then observed cell growth overlay area.The elongation nature of FBAM was tested on electronic universal testing machine by calculating elastic modulus,using bilayer amniotic membrane(BAM)and monolayer amniotic membrane(MAM)as controls.Results The fresh FBAM was integrated as a whole,with structurally complete fibrin and amniotic membrane as well as tight binding of fibrin to amniotic membrane.The reserved FBAM was structurally complete,brittler than the fresh FBAM,while amniotic membrane still bound tightly to the fibrin,but under microscope the fiber reticular formation was fairly fuzzy.Single layer of epithelial cells was formed on FBAM after about seven-day culture in vitro.Disparity of cell growth overlay area was significant(P

[Objective] To synthesize an amino acid coordinately compounds-Ferrous Glycine Sulfate(FeGly2) using Fe2+ as central iron and study the effect of blood enrichment.[Method]To synthesize the chelate iron using glycin and Fe2+ and measure the constructor of Ferrous Glycine Sulfate by IR spectrum and atomic absorption spectroscopy.To observe the blood enrichment effect of FeGly2 by rat anemia model.[Result] After separately treatment with 200 mg/kg FeGly2 and 300 mg/kg FeSO4 by intragastric administration,the haemoglobin in blood of hypoferric anemia rat increased obviously(P