OBJECTIVETo analyze the influence of retinoic acid (RA) on the undifferentiated state and EB formation abilities of human embryonic stem cells.

METHODSThe biological characteristics of H9 ESCs after RA treatment were characterized by real-time PCR, MTS proliferation assay and immunofluorescence staining. The expression of three germ layers markers, osteogenic differentiation markers and adipogenic differentiation markers in H9-differentiated embryoid bodies (EBs) with RA treatment were quantified by real time PCR.

RESULTSThe proliferation of H9 ESCs in the early logarithmic growth phase was accelerated by RA treatment. In addition, RA induced differentiation of H9 ESC coupled with morphology changes, decreased expression of undifferentiated markers Oct4, Nanog, Sox2 and OCT4 mRNA binding protein Lin28 at mRNA level, and reduced expression of Oct4 at protein level. RA induced formation of cavities in EBs. Real time PCR results showed that the expressions of ectodermal markers: NeuroD1, Noggin; mesodermal markers: Brachyury, Twist and endodermal markers: AFP, GATA-4 were significantly increased (P < 0.05), especially for AFP (P < 0.01), by RA treatment in a dose-dependent manner. In addition, the expression of adipogenic differentiation marker C/EBPalpha was increased while the osteogenic differentiation marker OPN was decreased in EBs after RA treatment for 5 days.

CONCLUSIONSHigh concentrations of RA induced the loss of stemness in H9 ESCs and excessive differentiation in EBs, and damaged the balance between osteogenic and adipogenic differentiation during early EB differentiation, which may be relevant to the congenital malformations.

Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; drug effects ; Humans ; Tretinoin ; pharmacology

Objective The hsa-miR-106b-25 gene cluster is involved in various biological processes of carcinoma .This study aims at a prediction and function analysis of the hsa-miR-106b-25 gene cluster, miR-106b, miR-93, and miR-25, so as to provide some evidence for further studies on the functions of the three miRNAs and the mechanisms of their interaction . Methods We obtained the sequences of miR-106b, miR-93, and miR-25 from the miRBase, predicted their target genes with TargetScan , PicTar, and miRanda, and used 3 or more experimentally verified target genes from the miRTarbase as the gene set for further bioinformatic analysis .We predic-ted the biological processes of the target genes by GeneOntology analysis and enriched KEGG ( Kyoto Encyclopedia of Genes and Genomes) by pathway analysis, produced protein-protein interaction ( PPI) networks with STRING . Results The target genes of the miR-106b-25 gene cluster were significantly enriched in such biological processes as the regulation of macromolecule metabolism , regulation of metabolic process , and cell cycle process , while the KEGG pathway mainly in glioma, melanoma, prostate cancer , and gallbladder carcino-ma.The proteins encoded by the targeted genes of showed complicated interactions , and those encoded by the KAT2B, PTEN, TP53, CDH1, MDM2, E2F1, RB1, and SMAD7 plaid a core role in the interac-tion network. Conc lusoi n MiRNAs of the miR-106b-25 gene cluster regulate the downstream target proteins involved in tumorigenesis by participating in the cell cycle and cancer signaling pathway .

Objective To investigate the effect of astaxanthin( ASX)on endothelial progenitor cells( EPCs)injury induced by oxidative stress in vitro and to explore its underlying mechanism. Methods Cultured EPCs isolated from peripheral blood were randomly divided into 5 groups:normal control,model group[ tert-butyl hydroperoxide( tBHP)100μmol·L-1 ],and ASX+tBHPgroups(thecellswerepreconditionedwithASX0.1,1.0,and10.0nmol·L-1,respectively).Thecellviabilitywas measured by MTT method. The level of reactive oxygen species( ROS)was determined by DCFH-DA method. The changes of mitochondrial membrane potential( MMP)and apoptosis ratio were detected by JC-1 method and DAPI method,respectively. caspase-3 activity changes of EPCs were detected. Results The cell viability of EPCs was improved with the increasing concentration of ASX. Compared with the model group[(48. 5±4. 3)%],0. 1,1. 0,10. 0 nmol·L-1 ASX significantly increased the cell viabilities[(57. 6±8. 2)%,(77. 6±7. 5)%,and(85. 3±6. 1)%,P﹤0. 05]. The results of DAPI staining revealed that ASX pretreatment could significantly reduce the apoptotic rate of EPCs. The apoptotic rate of the model group was( 27. 8 ± 3. 2)%,while that of ASX+tBHP groups was[(20. 4±2. 9)%,(14. 9±1. 7)%,and(7. 8±0. 7)%,P﹤0. 05],respectively. The data from caspase-3 activity assay indicated that ASX precondition could also remarkably decrease the caspase-3 activity for EPCs. The caspase-3 activity of the model group was(0. 345±0. 018),while that of the ASX+tBHP group were[(0. 291± 0. 013),(0. 209±0. 004),and(0. 169±0. 013),P﹤0. 05],respectively. In addition,treatment with tBHP resulted in an increase of DCF fluorescence,while ASX precondition could decrease the DCF fluorescence,which suggested the accumulation of intercellular ROS for EPCs. Injury of michondrial membrane resulted in the loss of mitochondrial membrane potential( MMP). The MMP detected by JC-1 method revealed that compared with model group,pretreatment of ASX inversed the reduction of MMP. Conclusion Astaxanthin inhibits endothelial progenitor cell apoptosis induced by oxidative stress through inhibiting ROS production,improving the mitochondrial function and down-regulating caspase-3 activity.

Objective To synthetically evaluate the relationship between miR-31 and the prognosis of carcinoma and to investi-gate its related mechanism.Methods The correlative literatures of tumor prognosis were retrieved from the electronic databases PubMed,EMBASE and ISI Web of Science.The pooled hazard ratios (HRs)and 95% confidence interval(95%CI )were extracted. The prognostic data were performed the synthesis analysis.Results A total of 7 trials conformed to the inclusion criteria including accumulated 2 012 cases of carcinoma.Meta-analysis revealed that the decrease of miR-31 expression in the tumor patients had the poor prognosis (HR=0.784,95%CI :0.630-0.974);in the subgroup analysis,the synthesis results adopting the multivariable a-nalysis and China subjects were 3.512 (95%CI :1.797-6.865)and 1.574 (95%CI :1.062-2.333),which indicating that the in-crease of miR-31 expression predicted the poor prognosis;miR-31 had no statistical significance in the digestive system (P >0.05). Conclusion The prognostic role of miR-31 may possess the histological and regional specificity and has the potential as a novel marker.

Objective To explore the technique,efficacy and complications of external sphincterotomy with Holmium laser for the treatment of detrusor external sphincter dyssynergia (DSD). Methods 7 male cases with neurogenic DSD who received the procedure were reported. Results All patients got significant improvement after the operation in symptoms,laboratory,imaging and urodynamic index. No one required transfusion. 1 case lost the erection. There was no need of re-operation during the follow-up. Conclusion External sphincterotomy with Holmium laser for the treatment of DSD shows the advantages of less bleeding,safety and good effectiveness for the well selected patients.

Objective To investigate the effect of Holmium laser urethrotomy under ureteroscopy on urethral stricture.Methods 30 men with urethral stricture who accepted urethrotomy with Holmium laser under ureteroscopy were observed.Results Operation was performed successfully in 21 cases;7 cases underwent 2～3 endoscopic surgical treatments.Endoscopic surgical treatment failed in 2 cases,and open surgery were performed on them.21 cases were followed up for 3～37 months,and 8 of them need urethral dilatation termly.Conclusion Endoscopic surgery with ureteroscopy and Holmium laser may be effective on urethral stricture with slight trauma.

Objective To explore the technique, efficacy and complications of transurethral sphincterotomy with Holmium laser for the treatment of male patients with neurogenic bladder and examine its long-term outcome. Methods 20 male cases who received transurethral sphincterotomy with Holmium laser were included in this retrospective analysis, in which 7 cases injuried at cervical spinal cord, 6 at thoracic spinal cord, 3 at cauda equine; 1 was spinal meningocele, 1 was sacral crack, 1 was ependymoma, and 1 was myelitis sequel. The age was 20~64 years, averaged 45.1 years. They were followed up for 3~95 months (averaged 43.6 months) since the last sphincterotomy. Results The 20 patients significantly improved in symptoms, laboratory examination, imaging demonstration and urodynamic index after the operation.There were 3 patients who required repeated laser surgery during the follow- up. 17 patients were stable. Conclusion Transurethral sphincterotomy with Holmium laser is effective on neurogenic bladder with less bleeding for the appropriately selected patients.

Objective To evaluate the effectiveness and safety of Botulinum toxin-A (BTX-A) injection into detrusor to treat neurogenic detrusor overactivity in patients with spinal cord injury (SCI).Methods A total of 78 patients with SCI were treated with transurethral injection of BTX-A (300 IU dissolved in 15 ml of saline) into 30 different points of detrusor with 15 ml in every patients. Urodynamic parameters and voiding diary were assessed at baseline and 3 weeks and 3 months after the injections. Adverse events were recorded after the injection if present.Results After the first injection, 78 patients showed that the mean frequencies of incontinence decreased from 13.5 to 2.7 times per day, the mean volume of intermittent catheterization (IC) increased from 131 ml to 389 ml per time, the mean volume of incontinence decreased from 1 690 ml to 281 ml per day, the mean getting effect time was 7.6 days. 10 patients received second injection at 8.9 months after first injection, the results showed that the mean frequencies of incontinence decreased from 9.7 to 3.7 times per day, the mean IC volume increased from 108 ml to 387 ml. 6 patients received third injection at 5.8 months after second injection, the results showed that the mean frequencies of incontinence decreased from 9.2 to 3.9 times per day, the mean IC volume increased from 116 ml to 364 ml. No side effects were observed during the follow-up.Conclusion BTX-A injection into detrusor to treat neurogenic detrusor overactivity in patients with SCI seems to be an effective, safe and miniinvasive solution.

BACKGROUND:Coronary stent implantation can cause blood vessel damage and wal reconstruction, leading to vascular stent restenosis. Studies have found that visfatin is associated with inflammatory reaction, and exhibits an increased expression at the site of plaque rupture in acute myocardial infarction. OBJECTIVE:To investigate the influence of percutaneous coronary intervention on the levels of visfatin in patients with coronary heart disease. METHODS:Thirty patients with acute myocardial infarction within 12 hours after the onset of the chest pain, 30 patients with unstable pectoris and 30 patients with stable angina pectoris were included. Al patients were successfuly treated by percutaneous coronary intervention. Meanwhile, 30 patients only undergoing coronary angiography but not stenting treatment were selected, and another 30 patients without any treatment served as normal control group. RESULTS AND CONCLUSION:According to enzyme-linked immunosorbent method, the visfatin levels of acute myocardial infarction, unstable angina, stable angina and coronary angiography groups continue to rise at pre-operation, 30 minutes, 6 hours, 12 hours, 24 hours after operation, al of which were higher than that in the normal control group (P < 0.05). The results confirmed that within 24 hours after coronary stent implantation the visfatin levels continue to rise.

Objective To investigate the value of endovascular surgery for the treatment of thromboangitis obliterans (TAO). Methods Sixteen patients (18 limbs) with thromboangitis obliterans were treated by percutaneous transluminal plasty (PTA). Results Angiographic success on the limb basis was achieved in 15 of 18 treated limbs and the initial technical success rate was 83.34%. Doppler anklebrachial index (ABI) increased from 0.33±0.16 to 0.79±0.23 one week after PTA. The follow-up time was 2～24 months, the average time was 10.84 months. The 3-month, 12-month accumulated primary patency rate were 81.33% and 60. 23% respectively. Conclusion PTA is effective primary invasive treatment for thromboangitis obliterans yielding acceptable primary clinical success with a low complication rate and resulting in moderate long-term clinical patency and a high limb salvage rate.