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.

Objective To observe the changes of four quantitative indexes of diabetic macular edema by using optical coherence tomography.Methods Eighty-nine patients (155 eyes) with diabetic retinopathy were included in this project and were divided into two groups according to the present of diabetic macular edema:Negative group (56 cases,100 eyes) and positive group(33 cases,55 eyes).In addition,23 cases (42 eyes) of normal volunteers constituted the normal control group.All the objects accepted an optical coherence tomography examination and the indexes including central retinal thickness (CRT),subfoveal choroidal thickness (SFCT) and integrity of external limiting membrane(ELM) as well as inner/outer segment (IS/OS) were measured and analyzed.Results The average CRT of positive group (219.048 ± 16.798) μm was significantly thicker than that of control group(217.775 ± 26.866) μm and negative group (280.418 ±74.187) μm (P ＜0.001).Mean SFCT among control group (312.893 ±140.559) μm,negative group (302.080 ± 125.287) μm and positive group (293.745 ±140.517) μm had no statistical significance (P =0.781).There were 3 eyes with disrupted ELM layer in the negative group and 8 eyes in the positive group.Difference between them was proved to be significant (P =0.019).Similarly,the integrity of IS/OS layer had significant difference between negative group (5 eyes disrupted) and positive group (19 eyes disrupted) (P ＜ 0.001).Conelusion CRT of patients with diabetic macular edema is always increased and the integrity of ELM or (and) IS/OS can be disrupted in many cases.Indexes including CRT and the integrity of ELM or (and) IS/OS can be used to evaluate the severity of diabetic macular edema quantificationally.

Objective To evaluate the clinical efficacy and safety of application of anterograde flexible ureteroscope in the treatment of ureterointestinal anastomotic strictures in patients after Bricker urinary diversion. Methods From March 2009 to July 2012, 6 patients with ureterointestinal anastomotic strictures after Bricker procedure were enrolled in this study. The average age of the patients was (61 ±7) years old. The first clinical presentation was averagely (6.3 ±2.5) months after the Bricker procedure. There were 4 cases with left side strictures and 2 cases with right side ones. The urinary tract ultrasound, CT, KUB+IVP and antegrade urography were carried out to identify the obstructive portion. The mean length of stricture was 0.9cm (0.4~2.5) . First, all patients underwent percutaneous nephrostomy (PCN), then inside incision by Holmium:YAG laser under anterograde flexible ureteroscopy and lithotripsy (with calculi) . The F6 double J ureteral stent was indwelled for 12 weeks. KUB+IVP was performed after removal of double J ureteral stents. Results The mean operative time was (53±8) min. The mean hospital stay was (5.5±2) days. The blood loss was 3~6 mL. The average follow-up was 18 months (6~30) . No recurrence was found in 5 patients. One case had recurrent stricture after the first procedure, which was successfully managed by the flexible ureteroscopy again and replacing double J ureteral stent every 12 months. Conclusion The inside incision by anterograde flexible ureteroscopic Holmium:YAG laser is safe and effective for ureterointestinal anastomotic strictures in patients after Bricker urinary diversion, with less complications.

Objective: To provide an important tool for the study of diagnose and treatment of prostate cancer (PCa) osseous metastasis and change of bone stress force on prostate cancer (PCa) osseous metastasis and a platform, which is more congruous to clinical process, for prevention and cure of neoplastic bone metastases, and to carry out the construction and improvement of animal models of PCa with different positional osseous metastasis in vivo.Methods: Different gradient concentrations of RM-1 cells were inoculated into the cavity of left femoral bone or lumbar vertebra of mice (C57BL/6) respectively.The change of mouse activity, tumor formation, tumor size and survival time were observed respectively.And the femur tissue and spinal tissue were obtained from the mice after death.The gray value of iconography were measured by imageological examination of femur tissue, and the final histopathological examination were taken to determine the tumor type in both femur and spinal tissue.Results: The tumor growth could be touched at the puncture site in all the mice after inoculated for 7 days.There were no obvious differences in the time of tumorigenesis, the rate of tumor growth and tumor size among the mice in the same group (P>0.05).As the result, the construction femoral bone and lumbar vertebra metastatic models of PCa had been confirmed by iconography and pathology detection.At the same time, the survival time of the mice inoculated with low concentrations of PCa cells was obviously longer than that of high concentrations of PCa cells (at least 2 weeks longer).Conclusion: The animal models with different positional osseous metastasis (limbs and axial skeleton) of PCa using the same PCa cells (RM-1) had been first constructed successfully in our study.At the same time, a high success rate of construction of PCa animal model with bone metastasis was obtained by femoral bone marrow cavity injection of PCa cells.The rate of tumor growth was rapid, animal survival time was appropriate, and the PCa animal model with bone metastasis can be stably reproduced by our method.These animal models can be used to explore the pathogenesis of different positional PCa bone metastasis and provide a new platform, which were more congruous to clinical process, for prevention and cure of neoplastic bone metastases.

Optical coherence tomography angiography (OCTA) base on OCT with an algorithm that can image a high-resolution picture of retinal circulation. OCTA has allowed quantifying the characteristic lesions of diabetic retinopathy (DR) in early stage, such as fovea avascular zone, retinal vascular density and the counts of retinal microaneurysm. In addition, OCTA can objectively evaluate the progression and prognosis of DR in late stage through imaging involved retinal neovascularization. Understanding OCT angiography features of DR lesions with different course of the disease may provide reference value for the diagnosis and treatment of DR.

Objective To observe the Th17 and regulatory T cells(Tr) equilibrium state as well as their changes of tuberculosis patients in six-month's anti-Tuberculosis treatment.Methods Select thirty-two tuberculosis patients received anti-Tuberculosis treatment while thirty-two healthy volunteers as controls.Flow cytometry was used to analyze Th17 and Tr cells in venous blood at the time of pre-therapy,3th,6th month.Results The ratio of Th17 cells in CD4 cells in tuberculosis patients and volunteers were (1.10±0.39)％,(2.50±1.03) ％,(3.90±1.34) ％,(4.50±1.52)％,respectively; the ratio of Tr cells were (9.17±3.26)％,(6.85±2.73)％,(5.46±1.69)％,(4.35±0.86)％,respectively.Conclusion Tuberculosis could make Th17 cells and Tr cells lost their balance,but the immune equilibrium state may gradually recover after anti-tuberculosis.The change of the amount of immune cells was likely to be the reference indexes to observe the progress of tuberculosis and the treatment effect of anti-tuberculosis.

Objective To design and synthesize novel triazole antifungal derivatives with 1 ,3 ,4-oxadiazole side chain for the study of antifungal activities. Methods Fourteen title compounds were synthesized via acylation ,aminolysis reaction ,cy-clization ,nucleophilic substitution ,etc. All the compounds were characterized by 1 H NMR ,MS spectra. The in vitro antifun-gal activities were evaluated against six human pathogenic fungi through the micro-broth dilution method. Results The title compounds exhibited strong antifungal activities against all the tested fungi ,especially against Candida albicans. Compounds 10d ,10i , 10l , and 10n were found to be the most effective , with a minimum inhibitory concentration (MIC80 ) of 0.003 9 μg/ml .They are 16-fold more potent than ICZ ( MIC80 0.062 5 μg/ml) and 64-fold more potent than FCZ (MIC80 0.25 μg/ml) .Conclusion The 1 ,3 ,4-oxadiazole side chain could affect the antifungal activities. That could be due to the prop-er incorporation between the 1 ,3 ,4-oxadiazole substituted phenyl ring with the target enzyme.

Objective: To study the oxidative damage of di- (2-ethylhexyl) phthalate (DEHP) on MCF-7 cells, and to investigate the effects of 3β-hydroxysteroid dehydrogenase (3β-HSD) gene silence or overexpression on DEHP-induced oxidative damage. Methods: MCF-7 cells, 3β-HSD gene silencing cells and 3β-HSD gene overexpression cells were treated with different doses of DEHP (0,0.05,0.1,0.2,0.4,0.8 mmol/L) for 24h, then intracellular oxidative damage index such as MDA, SOD, GSH, GSH-PX were detected, DNA repair gene hOGG1, hMTH1 mRNA expression were tested by Q-PCR, hOGG1, hMTH1 protein expression were detected by western blot. Results: After MCF-7 cells were treated by DEHP, MDA levels increased; SOD activity, GSH content, GSH-PX activity decreased, hOGG1 and hMTH1 mRNA expression levels increased, hOGG1 and hMTH1 protein expression levels increased, the differences were statistically significant when compared with control (P<0.05 or P<0.01) . In 3β-HSD gene silencing cells which were treated by DEHP, when compared with the same dose group of MCF-7 cells, MDA content increased, SOD activity, GSH content, GSH-PX activity decreased, hOGG1 and hMTH1 mRNA expression levels decreased, hOGG1 and hMTH1 protein expression levels decreased, the difference were statistically significant (P<0.05 or P<0.01) . In 3β-HSD gene overexpression cells which were treated by DEHP, when compared with the same dose group of MCF-7 cells, MDA content decreased; SOD activity, GSH content, GSH-PX activity increased, of hOGG1 and hMTH1 mRNA expression levels increased, hOGG1 and hMTH1 protein expression levels increased, the difference were statistically significant (P<0.05 or P<0.01) . Conclusion DEHP could cause oxidative damage in MCF-7 cells, induce the changes of related genes and proteins, 3β-HSD plays an antioxidant role in the process of DEHP ox-idative damage.

OBJECTIVE: To investigate the effect and mechanism of glucagon like peptide 1(GLP-1)on the proliferation and differentiation of endothelial progenitor cells(EPCs)derived from the peripheral blood. METHODS: Mononuclear cells were isolated from human peripheral blood by density gradient centrifugation. After 7 days of culture,attached cells were stimulated with different cultures of 0.2% BSA,and GLP-1(1,10,and 20 nmol/L). Laser scanning confocal microscope was used to determine the EPCs from human peripheral blood.The activity of EPCs was observed under reverse microscope. MTT was used to determine the proliferation of EPCs. The expression of KDR,Flt-1,VE-cadherin,and eNOS mRNA was detected by RT-PCR.The concentration of serum VEGF was detected by ELISA. The expression of VEGF protein was detected by immunohistochemical SP method. The EPCs cultured in GLP-1 were intervened by VEGFmAb. RESULTS: EPCs was proliferated more in the GLP-1 group(1,10,and 20 nmol/L) than in the control group (P<0.05 or P<0.01). The expression of KDR,FLT-1,VE-cadherin,eNOS mRNA and VEGF protein was higher than that in the control group(P<0.05 or P<0.01). VEGFmAb(100 ng/mL)down-regulated the expression of KDR,Flt-1,VE-cadherin,and eNOS mRNA. CONCLUSION GLP-1 can promote the proliferation and differentiation of EPCs derived from the peripheral blood by up-regulating VEGF autocrine.
Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Endothelial Cells ; cytology ; Glucagon-Like Peptide 1 ; pharmacology ; Humans ; Leukocytes, Mononuclear ; cytology ; Stem Cells ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Objective:To investigate the protective effect of quercetin(QUE)on acute lung injury(ALI)rats with sepsis.Methods:Sixty male SD rats were randomly divided into 6 groups(n=10):Sham operation group(Sham),model group(CLP),QUE 25 mg/kg group,QUE 50 mg/kg group,QUE 100 mg/kg group and positive drug dexamethasone(DEX)group.Rats in each group were continuously treated for 7 days,and the survival rate was calculated;HE staining and lung wet-to-dry weight ratio(W/D)were used to evaluate the severity of lung injury;TUNEL staining was used to detect lung tissue apoptosis;levels of inflammatory factors,SOD and MDA were detected by the kits;Western blot was used to detect expressions of apoptosis-related proteins and phosphoryla-tion levels of PTEN,β-catenin,protein kinase B(AKT)and glycogen synthase kinase-3β(GSK-3β)in rat lung tissue.Results:Com-pared with CLP group,after QUE 50 mg/kg and 100 mg/kg treatment,survival rate of rats was significantly increased(P<0.05),the degree of inflammatory cell infiltration in lung tissue was reduced,lung injury score and W/D were reduced(P<0.05),apoptosis rate of lung tissue cells and expressions of apoptosis-related proteins were significantly decreased(P<0.05),levels of inflammatory factors were decreased(P<0.05),while the antioxidant capacity was enhanced(P<0.05),phosphorylation levels of PTEN and β-catenin were decreased,while phosphorylation levels of AKT and GSK-3β were increased(P<0.05).Conclusion:QUE protects rats from ALI with sepsis by inhibiting apoptosis,reducing inflammation and antioxidance,which mechanism may be related to PTEN/β-catenin and AKT/GSK-3β pathways.