MicroRNAs are endogenously expressed non-coding RNAs, which are composed of approximately 18 nucleotides to 25 nucleotides. Mature microRNAs regulate gene expression by base pairing with the 3'-untranslated region of target mRNAs. These mature microRNAs can degrade target mRNAs or inhibit translation. This process is a type of post-transcriptional regulation of gene ex-pression. Studies have shown that microRNAs are important in physiological and pathological processes, such as cell proliferation, cell differentiation, and cell death. This article provides an overview of the function of microRNAs in the regulation of macrophages.

The aim of this study was to synthesize 1-(5-(hydroxymethyl)-4-methyl-3-oxo-3,4-dihydro-pyrazin-2-yl) guanidine (mucunaguanide),a pyrazinone alkaloid compound extracted from the seed of Stizotobium cochinchinensis.Starting from benzyloxy acetaldehyde,following four steps in reaction including condensation,cyclization,substitution and hydrogenation reaction,mucunaguanide was synthesized with total yield of 27.9％,and purity of more than 97.5％.Structures of all the intermediates and target compound were confirmed by IR,1H NMR and MS.This synthetic process was characterized with raw materials available,simple in operation with much milder reaction conditions,and was an ideal method of synthesis of this compound.

ObjectiveTo investigate the effects of autologous bone marrow-derived mesenchymal stem cells (MSCs)transplantation on acute myocardial infarction in swine models using MRI. MethodsFourteen Chinese mini-pigs(27±3 kg)were divided into control group(n=7)and transplantation group(n=7).Acute myocardial infarction(AMI)model was made by occlusion of the left anterior descending coronary artery for 90 minutes,and then 10 ml autologous MSCs(3 × 106 cell/ml)were injected into LAD by over-wire-balloon catheter after one week. MRl was performed to assess the cardiac function and myocardial perfusion 1 week after AMI and 6 weeks after transplantation.The implanted cells in vitro were analyzed by immunofluorescence.ResuitsThe left ventricular ejection fraction(LVEF)in transplantation group was increased from(42.7 ±7.5)%to(50.1±10.1)%,which was significantly different from that in control group(P＜0.01).In addition,the dyskinetic segments in infarcted region and the infareted area were decreased by 4 and 3.2 cm2 respectively(P＜0.01),and the left ventricular weight index was increased by 4.1 g/m2 in transplantation group(P＜0.05)compared with control group.The DAPI-labeled cells in infarcted and peri-infarcted region indicated the survived MSCs.Immunofluoreseence also confirmed that those cells expressed cardiomyocyte-specific troponin T,connexin 43 and vessel-specific smooth muscle actin.Capillary density in both infarcted and peri-infarcted region were higher in transplantation group than the control group(P＜0.01).Conclusion MRI is a reliable imaging method for assessing the effects of stem cell transplantation in acute myocardial infartion of swine models.

10.3969/j.issn.2095-4344.2013.25.023

Objective To investigate the effect of valproic acid (VPA) on NKG2D-ligand expression in ARK,OPM2 human myeloma cell lines and their sensitization to natural killer (NK) cell-mediated Killing.Methods Different concentrations of VPA from 0-5.0 mmol/L were used to treat ARK,OPM2 cells respectively,then the cell viabilities were tested by flow cytometry (FCM).Real-time quantitative-PCR and FCM were used to detect the changes in mRNA,protein levels of NKG2D-ligand respectively in the two cell lines treated with 1 mmol/L VPA for 48 hours.The calcein-release-assay (CARE-LASS) was carried out to detect cytotoxic changes of NK cells against mydoma cells after VPA treatment.Results VPA induced the expression of MICA/B,ULBP2 (P ＜ 0.05) and in turn enhanced the NK cytotoxicity on myeloma cells.The enhancing effect of VPA was blocked by NK cells pretreated with anti-NKG2D mAb (P ＜ 0.05).The primary mechanism of NK cell killing of myeloma cells was perforin/granzyme-mediated.Conclusion VPA can induce the expression of MICA/B,ULBP2 in ARK,OPM2 cells,thereby enhancing the cytotoxicity against myeloma cells,which implies a new mechanism of anticancer approach and may be a new approach in myeloma immunotherapy.

Objective To evaluate the clinical value of the combination of S-shaped urethral dilator and nephro-scope for the treatment of male urethral stricture disease. Methods Guidewires were inserted into bladder through the nephroscope under direct vision. The urethral dilation with S-shaped urethral dilator was carried out by nephroscope in 41 male patients. All catheters were located across the strictures and remained for 4-6 weeks. The regular follow-up was done on all cases to assess the clinical effect on urine flow. Results All surgeries were successful without serious complications. The mean operative time was (40.16 ± 5.78) min. The voiding symptoms were significantly improved after catheter removal compared with those of preoperation in all cases. Patients were followed up after surgery and were regularly urethral dilation at least 6 weeks. The mean follow-up time was (34.75±6.42) weeks.There were incontinence, diminished sexual function and other complications after operation in all cases. Conclusion Nephroscope combined with S-shaped urethral dilator for the treatment of male urethral stricture disease is feasible, minimally invasive and safe, which is worthy of recommendation.

BACKGROUND: Vessel extracellular matrix (VECM) is a natural scaffold material obtained from vascular tissues, which can stimulate angiogenesis and accelerate vascularization of tissue-engineered graft, however, the mechanism is poorly understood.OBJECTIVE: To explore the vascularization effects of release of vascular endothelial growth factor (VEGF) from VECM in ureteral reconstitution.DESIGN, TIME AND SETTING: An in vitro cytology observation. The experiment was performed at the Biomedical Engineering Laboratory of First Clinical Medical Science College, Wuhan University, between April and August in 2009.MATERIALS: Abdominal aorta was obtained from 5 rabbits to prepare VECM.METHODS: The VEGF released from VECM in vitro was evaluated by ELISA, the effects of cell proliferation by the released VEGF was detected by MTT colorimetric assay. The defected ureters of rabbits were repaired by homologous VECM in vivo.Then the recovery of the defected ureters and the situation of vasculogenesis were detected at different time point.MAIN OUTCOME MEASURES: The detection of VEGF contents in VECM; and the effects of VECM on vascular endothelial cell proliferation and ureteral reconstitution.RESULTS: In vitro experiment presented that the peak amplitude concentration of VEGF released from VECM in PBS solution was (124.10±1.42) ng/L, which showed proliferative effect on vascular endothelial cells. In vivo, there were some blood vessels on the VECM at 2 weeks after implantation. Epithelial coverage was evident in the lumen of the marginal part of the VECM grafts and the smooth muscle extended from the transition zone. After 8 weeks, the quantity of the blood vessel was increased and the caliber of the blood vessels became wide. There was thickness epithelial lamina in the graft, and the muscle fibers had an organized spatial alignment, forming variably sized bundles. After 16 weeks, there were no significant differences between the regenerative tissue and the normal tissue in morphology.CONCLUSION: The homologous VECM can release VEGF when implanted as tissue engineer biomaterial and might be an ideal replacement biomaterial for ureteral reconstitution.

Objective To investigate the effects of salinomycin on the cell proliferation and epithelial-mesenchymal transition (EMT) of MCF-7 mammosphere (MCF-7 MS). Methods Breast cancer MCF-7 cells were cultured in suspension in serum-free medium to obtain MCF-7 MS. The cell viability of MCF-7 MS cells treated with serial concentrations of 0, 10, 30, 100, 300, 1 000, 3 000 and 10 000 nmol/L of salinomycin for 24 hours were detected by CCK-8 assay. The half maximal inhibitory concentration (IC50) was calculated. Western blot analysis was performed to detect the expression levels of E-cadherin and Snail in MCF-7 MS cells treated with 30 nmol/L and 60 nmol/L salinomycin. The same capacity of DMSO was added to MCF-7 MS as control group. The xenograft tumors from MCF-7 MS transplant mice were divided into control group (the same capacity of normal saline) and salinomycin group (5 mg/kg salinomycin), then the expressions of E-cadherin and Snail were dectected by immunohistochemical staining. Results With the increased concentration of salinomycin, the cell survival rate of MCF-7 MS cells decreased (P<0.05). The IC50 after 24 h-treatment was 989 nmol/L. Both 30 and 60 nmol/L of salinomycin increased the expression of E-cadherin and decreased the expression of Snail compared with control group. In addition, 60 nmol/L treatment group showed more significant effect (P<0.05). In xenograft tumors from MCF-7 MS transplant mice, the expression of Snail decreased, and E-cadherin increased in salinomycin treatment group compared with control group (P<0.01). Conclusion Salinomycin can inhibit the cell proliferation and EMT in MCF-7 MS cells, which is a potential drug to target cancer stem cells.

ObjectiveTo explore the value of conventional echocardiography and transthoracic real-time three-dimensional echocardiography (RT3DE) in diagnosis of Ebstein anomaly.MethodsWe investigated the morphology and function of right ventricle (RV) as well as the structure, morphology and regurgitation of tricuspid valve in 61 adult patients with Ebstein anomaly before surgery by using conventional echocardiography and transthoracic RT3DE. Twenty normal adults were enrolled as control group. ResultsThe surface of the tricuspid leaflets, the morphology of the tricuspid annulus as well as the three-dimensional structure of the tricuspid valve were displayed stereoscopically by RT3DE. Complete RV volume data could be acquired in 32 patients of Ebstein anomaly. The apex or part of RV could not be contained in the remaining 29 patients. Thirty-four (55.7%) patients with Ebstein anomaly had severe tricuspid regurgitation,
16 (26.2%) patients had moderate to severe regurgitation, and 11 (18.1%) patients had moderate regurgitation. Compared with the normal adults, patients of Ebstein anomaly showed higher RV end-diastolic volume (EDV), end-systolic volume (ESV), end-diastolic volume index (EDVI), end-systolic volume index (ESVI), stroke volume (SV), RV anterior-posterior diameter (RV), tricuspid valve annular transverse diameter (TV-R), and lower RV ejection fraction(EF) [(273.5±77.7) mlvs (74.3±15.9) ml, (187.1±96.8) mlvs (31.1±9.2) ml, (177.4±53.6) ml/m2vs (43.4±8.2) ml/m2, (121.7±65.5) ml/m2vs (18.4±5.1) ml/m2, (95.9±20.2) ml vs (43.6±8.8) ml, (48.1±13.3) ml/m2vs (19.0±1.9) ml/m2, (56.4±8.9) ml/m2vs (28.5±4.3) ml/m2, (38.3±12.8) %vs (59.3±5.1) %, allP<0.05). The tricuspid regurgitant orifice flow cross-sectional area (EROA) were correlated positively with RV anterior-posterior diameter (r=0.691), ratio of RV and LV anterior-posterior diameter (RV/LV) (r=0.6471).ConclusionTransthoracic RT3DE is a feasible method in addition to conventional two-dimensional echocardiography in evaluation of tricuspid valve mophology and function, as well as RV volume and EF in adult patients with Ebstein anomaly.

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