BACKGROUND:Umbilical cord blood-derived mesenchymal stem cel s are multipotential stem cel s in the mesoderm in early development stage, and have been paid great attention due to its properties of multi-directional differentiation. OBJECTIVE:To summarize the potential of induced differentiation of umbilical cord blood-derived mesenchymal stem cel s. METHODS:We retrieved PubMed Database for articles concerning the differentiation potential of umbilical cord blood-derived mesenchymal stem cel s published from January 1999 to December 2012. In titles and abstracts, the key words were“umblical cord blood, mesenchymal stem cel s, potential, differentiation”. Total y, 52 articles addressing the differentiation potential of umbilical cord blood-derived mesenchymal stem cel s were reviewed. RESULTS AND CONCLUSION:Numerous studies have confirmed that human umbilical cord blood-derived mesenchymal stem cel s can successful y differentiate into multiple kinds of cel lines, but their understanding remains minor. If we can master the characteristics of the differentiation potential of umbilical cord blood-derived mesenchymal stem cel s, it would be used to repair bone and myocardium detects. Present studies remain in a starting stage. Isolation and purification, regulation of differentiation direction, in vitro amplification and immunogenicity require further investigations.

Chronic cough is a multi-factorial symptom,postnasal drip syndrome (PNDS) and gastro-esophageal reflux disease (GERD) are common causes of chronic cough, which is closely associated with the otolaryngologist. The aim of this paper is to highlight the issues in clinical features, diagnosis and management of chronic cough from the otolaryngologist perspective.
Chronic Disease ; Cough ; diagnosis ; therapy ; Humans

Objective To investigate the gene silencing,apoptosis induction and the suppression of proliferation in vivo transfected by UTMD techniques associated with shRNA techniques. Methods The survivin-shRNA expression vector was constructed. Nude mice were randomly arranged into 3 groups:control group, plasmid injection and ultrasound (P + US), P + UTMD group. Histological examination were evaluated. Protein expressions of Survivin and proliferating cell nuclear antigen (PCNA), Bcl-2, Bax,Caspase-3, Ki-67, nucleostemin (NS), p53 were investigated by immunohistochemistry. Results In transplanted tumors experiment, comparing with those in C and P + US groups, protein expressions of PCNA,Ki-67,Bcl-2, Survivin, NS were down-regulated markedly, while those of Bax, Caspase-3 and P53 were up-regulated significantly ( P ＜ 0.05). Conclusions UTMD combined with shRNA technique can induce apoptosis and inhibit proliferation significantly, without causing any apparently adverse effect,representing a new,promising technology that can be used in the tumor gene therapy and research.

Objective To improve the identification of rush pneumoconiosis.Methods To analyze 47 cases of rush pneumoconiosis treated in our hospital from 2003 May to 2008 December.Results The male occupied 74% of 47 patients.The average age of onset was 34.2 ranging from 26.1 to 42.3.78.7% patiets presented with cough,sputum production,chest pain and dyspnea.4.2% patiets had PaO2 45 mmHg.85.1% patients' chest X-ray and CT images showed nodule shadows,interstitial fibrosis and ground glass attenuation.42.6% patients had restrictive ventilation disorders,2.1% obstrutive ventilation disorders 4.2% mixed ventilation disorders,and 74.5% decreased diffusing capacities.Through fiberoptic bronchoscopy,carbon sediment were seen on the 10% patients' bronchial walls,and bronchial lumen were distorted and stiff.Chronic inflammaion increased,macrophages and fibre tissues,and 21.2% dust cells were seen in pathology.All patients were treated according to their clinic symptoms.Conclusion Rush-mat dust was the main cause of rush pneumoconiosis,there were no special ways to cure this disease,prevention was the key to eliminate rush pneumoconiosis.

Objective To explore the changes and significance of serum tumor necrosis factor-?(TNF-?),interleukin-1?(IL-1?),IL-6 and IL-10 in rats with acute necrotizing pancreatitis(ANP).Methods Sixty-four Wistar rats were randomly divided into 2 groups: sham operation group(SO group n=32) and ANP group(n=32).The ANP model was established by using retrograde injection of Sodium Taurocholate into cholangiopancreatic duct.The changes of serum endotoxin(ET),TNF-?,IL-1?,IL-6 and IL-10 in different groups and different time points were observed.Results The levels of serum ET,TNF-?,IL-1?,IL-6 and IL-10(except IL10 of ANP group in 16h) in rats of ANP group were significantly higher than those of SO group(P

Objective To study the transfeetion efficiency and safety of liposome microbubble(LM)on red fluorescent protein(RFP)in vitro and in vivo under ultrasound mediated gene transfection(USMGT)conditions.Methods Plasmids containing RFP were added to cultured Hela cells followed by ultrasound (US)exposure with LM.Different concentration of LM,US intensity and exposure time were optimized.Transfection efficiency was evaluated by fluorescent microscopy and FACS.Cell viability was verified by propidium iodide assay.In transplanted tumors in vivo study,LM and plasmid(P)were injected into the nude mice followed by US exposure(P+LM+US group).Nude mice undergoing plasmid injection alone(P group),plasmid injection and US exposure(P+US group)and plasmid and LM injection(P+LM group)were used as controls.Frozen section and histological examination were conducted and RFP expression was evaluated.Results LM and US exposure significantly increased transfeetion efficiency in cultured Hela cells (P＜ 0.01).Transfection efficiency was the most prominent under the condition of US intensity of 1.0 W/cm2 with 6%LM,duration 3 min.No apparent cell damage was found in the all groups.In transplanted tumors,strong RFP was seen in P+LM+US group.It was significantly higher than in any other groups(P＜0.0 1).No tissue damage was seen histologically.Conclusions LM could enhance USMGT effectively without causing any apparently adverse effect in vitro and in vivo.This method would be a novel,effective,safe non-viral gene transfection method and provide an alternative to current clinical gene therapy.

Objective To determine whether it could enhance gene delivery and tumor transfection in vivo by combination of ultrasound-targeted microbubble destruction(UTMD)with polyethylenimine(PEI)in tumor xenografts.Methods Two different reporter plasmid[lueiferase(pCMV-LUC)and red fluorescent protein(RFP)]were incubated with PEI to prepare cationic compound(PEI/DNA)in various nitrogen:phosphate ratios(N/P ratios,nmol of nitrogen in the PEI/nmol of phosphate in DNA).Formation of PEI/DNA complexes were confirmed by the gel retardation assay.Human cervical carcinoma(Hela)tumors were planted subcutaneously in both flanks of female nude mice.Tumor-bearing mice were administered by tail vein with PBS,plasmid,plasmid and Sono Vue microbubble,PEI/DNA and Sono Vue microbubble.One tumor was exposed to ultrasound irradiation (3 MHz,2 W/cm2,2 min exposure,duty cycle 20%),while the other served as control.The feasibility of targeted delivery and tissue specificity facilitated by UTMD and PEI was investigated.The mice were sacrificed 3 days after ultrasound exposure.Tissue specimens were viewed with microscopy to determine the presence of RFP expression.The efficiencies of luciferase transgene expression were determined.Histology analysis was detected.Results Electrophoresis experiment revealed that PEI was mixed with plasmid to condense DNA efficiently.The application of UTMD significantly increases the tissue transfection in vivo compared to plasmid alone.RFP expression was present in all sections of tumors that received ultrasound exposure but not in control tumors.Results of luciferase activity showed that the expression of luciferase was to be 14 times greater in ultrasound-exposed tumors(P＜0.01).More importantly,the increase in transgene expresgion was related to UTMD with the presence of PEI dramatically.At least 10-fold increase of luciferase gene transfer was obtained in irradiated tumors compared to non-irradiated controls(P＜0.01),111-fold increase compared to UTMD alone(P＜0.01).There was not significantly gene expression in other organs or tissues regardless of US exposure(P＞0.05).No tissue damage was seen histologically.Conclusions The combination of UTMD with PEI can enhance targeted delivery and expression of reporter gene to tumors at intravenous administration.It is a promising new and safe method for gene delivery in vivo.

Objective To investigate different ultrasound parameters and transfection conditions that would affect transfection rate of red fluorescent protein(RFP)and cell viability of cancer cells.Methods In this study,Hela cells were cultured using two different protocols:(A)24 h culture for complete adherence;(S)suspension.Subsequently,cells were transfected following different ultrasound exposure protocols[1.0W/cm2;duty cycle(DC):10%,20%and 50%;exposure 1min or 3 min].Gene transfection and cell viability were evaluated.Treatment parameters optimized in Hela cells were applied for delivery RFP in 4 other cell lines(HepG2,Ishikawa,MCF-7 and B16-F10).Results Cell injury were found to increase progressively with DC and exposure time in group A.Cell detachment was significantly accompanied by ultrasound exposure in adherent HeLa cells.Cells in group S were found more prone to be transfected than group A with the same ultrasound parameters,while the survival rate was not decreased apparently.The ideal ultrasound conditions were noted to be at 1.0 W/cm2 irradiated 3 min with 20%DC using suspended protocol,producing maximum efficiency[transfection=(28.04±2.27)%]in gene delivery with minimum cell toxicity[cell viability=(81.20±1.73)%].These experiments also revealed different response to ultrasound treatment,but for all tested cell lines,dead and transfected cells in the treated groups were significantly different from the non-irradiated groups.Conclusions Ultrasound parameters and transfection conditions have a great impact on the gene delivery and cell viability.Gene delivery of ultrasound-mediated microbubble enhance should be optimized to improve the efficiency.

Objective To study the optimized condition of transfection efficiency for MCF-7 cells enhanced by ultrasound(US) irradiation and contrast agent combined with polyethyleneimine(PEI) and observe whether the combination can have a synergistic effect to increase DNA transfection. Methods MCF-7 cells were transfected with the compounds prepared by the vector of plasmid DNA encoding luciferase (pCMV-luciferase-GL3) and PEI.SonoVue microbubble was added to the cell suspension to serve as nucleation sites for aeoustic cavitation before US irradiation. The DNA expression of luciferase plasmid and viability of cells were evaluated. The strategy of US irradiation was optimized. Furthermore, the influencing factor, such as the concentration of plasmid, incubation time, serum, the type of solvent and the volume of culture media, were examined. Results The viability of cells and US-induced enhancement of luciferase activity were influenced by the US intensity,exposure time and duty cycle.US irradiation under an appropriate condition enables ceils to accelerate the permeation of the PEI/DNA complex through the cell membrane, resulted in enhanced transfection efficiency of plasmid DNA. Optimal US condition for the enhancement was determined to be 1 W/cm2,10% DC for 3 min. In contrast to the PEI/DNA complex alone without US irradiation or US irradiation alone, the combination of US irradiation with contrast agent and PEI had a significantly enhanced luciferase activity (P<0.01). The 2 h pre-irradiation incubation with PEI/DNA complex for MCF-7 ceils exhibited a significantly enhanced lueiferase activity (P<0.01). Besides,serum,type of solvent and the volume of culture media did affect the transfection efficiency. Conclusions The optimized parameters of US and transfection provide efficient gene delivery in MCF-7 cancer cells. The combination of US irradiation with contrast agent and PEI has a synergistic effect to increase DNA transfection. This is a simple and promising method to enhance the gene expression of plasmid DNA.

Objective To investigate different pulsed ultrasound (PUS) parameters and culture conditionsthat would affect cell viability and sonoporation on cell membrane of human cervical cancer cells (HeLa). MethodsHeLa cells were cultured in two different conditions ( in suspension or in monolayer). Cells were exposed to differentPUS intensity (0.4 W/cm2, 1.0 W/cm2, 1.6 W/cm2, 2.2 W/cm2), duty cycle (10%, 20%, 50%) and expo-sure time ( 1 min or 3 min). Cell viability was analyzed by flow cytometry. Using microscope and scanning electronmicroscopy (SEM) , the changes of shape and the sonoporation on cell membrane induced by PUS were observed.Results Low intensity and duty cycle did not exert a great impact on the cell viability. Cell injury was found to in-crease progressively with high intensity ( 1.6 W/cm2 , 2.2 W/cm2 ) and duty cycle ( 50% ) ( P < 0. 01 ) , and celldetachment was significantly accompanied by PUS exposure in adherent HeLa cells. Results of factorial design showedthat the culture conditions and the PUS parameters had significant interaction ( P < 0.01 ). SEM demonstrated insome detail the phenomenon of transient pores in the cell membrane under suitable PUS irradiation. The ideal sonopo-ration conditions that cell viability was above 80% and more membrane holes were noted to be at 1.0 W/cm2 expo-sure for 3 min with a duty cycle of 20% in cell suspension. Conclusion The optimized conditions of the PUS pa-rameters and the culture conditions could lower the cell injury and exert a great impact on the sonoporation. It couldproduce remarkable membrane pores on cells and enhance cell membrane permeability, which facilitate transportationof macromolecules into cells.