Adenoidectomy ; Female ; Humans ; Infant ; Male ; Retrospective Studies ; Sleep Apnea, Obstructive ; surgery

Objective:To introduce a "Four-step method" for extraction and separation of Candida albicans total proteins.Methods: Proteins of C.albicans were extracted step-by-step with 4 kinds of solutions with different solubilities.After quantification,the protein samples were separated by isoelectric focusing electrophoresis and then by SDS-PAGE.Results: Proteins with different solubilities were successfully extracted step-by-step from C.albicans and were separated by two-dimensional gel electrophoresis. Conclusion: The "Four-step method" for extraction of C.albicans proteins is an effective approach to study C.albicans membrane proteome and lays a foundation for further investigation of the mechanisms of antifungal agents and drug resistance in C.albicans.

Objective To compare the efficacy and efficiency of simulation-based training of flexible fibreoptic intubation in novices with virtual reality simulator.Methods A total of 46 anaesthesia residents in their first stage of training in anaesthesiology with no experience in flexible fibreoptic intubation at Peking University People's Hospital were enrolled in the study,and were divided into 2 groups randomly,which were virtual reality simulator group (group S,n=23) and manikin group (group M,n=23).The group S was then trained for 25 times on simulator,while the group M did the same processes on manikin.After training,participants in both groups had their performance assessed with the fibrescope evaluated through the oral route using a simulation manikin,who were instructed to attempt to advance the fibrescope 5 consecutive times to view the carina in the shortest amount of time.The time required to view the carina of each practice during training in both groups were recorded as pooled data to construct group learning curves with the application of SPSS 20.0.By using repeated measures analysis of variance and Ttest,the procedure time and global rating scale (GRS) of fibreoptic bronchoscope manipulation ability were compared between groups,so did the participant's confidence between before and after the training both within-subjects and between-subjects.Results The plateaus in the learning curves were achieved after 19 (15,26) practice sessions in group S and 24 (19,31) in group M,respectively.There was no significant difference in the procedure time [(13.7 ± 6.6) s and (11.9 ±4.1) s] and GRS [(3.9 ±0.4) vs.(3.7 ±0.3)]between groups.There were significant increases in participant's confidences in both groups after training [group S:(1.8 ± 0.5) vs.(3.9 ± 0.6),t=10.928,P=0.000;group M:(2.0 ± 0.7) vs.(3.9 ± 0.5),t=15.306,P=0.000],but there was no significant difference between groups.Conclusion The simulation-based training of flexible fibreoptic intubation in novices with virtual reality simulator is more efficient than the one with manikin,but the similar effects can be achieved in both modalities,after adequate trainings.In the related training a balance between time cost and economic cost should be considered and the appropriate teaching methods and forms should be taken.

To establish a LC-MS/MS method to determine the concentrations of liquiritin, glycyrrhizin, glycyrrhetinic acid, amygdalin, amygdalin prunasin, ephedrine, pseudoephedrine and methylephedrine of Maxing Shigan decoction in rat plasma, and study the differences on their pharmacokinetic process in normal rats and RSV pneumonia model rats. After normal rats and RSV pneumonia model rats were orally administered with Maxing Shigan decoction, the blood was collected from retinal vein plexus of different time points. Specifically, tetrahydropalmatine was taken as internal standard for determining ephedrine, while chloramphenicol was taken as internal standard for determining other components. After plasma samples were pre-treated as the above, the supernatant was dried with nitrogen blowing concentrator and then redissolved with methylalcohol. The chromatography was eluted with mobile phase consisted of acetonitrile and 0.1% formic acid solution in a gradient manner. ESI sources were adopted to scan ingredients in ephedra in a positive ion scanning mode and other ingredientsin a negative ion scanning mode. The multiple-reaction monitoring (MRM) method was developed the plasma concentration of each active component. The pharmacokinetic parameters of each group were calculated by using Win-Nonlin 4.1 software and put into the statistical analysis. The result showed the plasma concentration of the eight active ingredients, i.e., liquiritin, glycyrrhizin, glycyrrhetinic acid, amygdalin, amygdalin prunasin, ephedrine, pseudoephedrine and methylephedrine within the ranges of 1.04-1040, 1.04-1040, 0.89-445, 1.05-4200, 1.25-2490, 0.3-480, 0.3-480, 0.3-480 microg x L(-1), with a good linearity and satisfactory precision, recovery and stability in the above ingredients. After modeling, except for glycyrrhetinic acid whose pharmacokinetic parameters were lacked due to the data missing, all of the rest components showed significant higher Cmax, AUC(0-1) and lower clearance rate (CL) than that of the normal group, indicating the increase in absorption in rats in the pathological state by reducing the clearance rate. The method is accurate and sensitive and so can be used to determine the plasma concentrations of the eight active ingredients in Maxing Shigan decoction. RSV pneumonia-infected rats absorbed more ingredients in Maxing Shigan decoction.
Animals ; Chromatography, High Pressure Liquid ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacokinetics ; Male ; Pneumonia, Viral ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Respiratory Syncytial Virus Infections ; drug therapy ; metabolism ; Tandem Mass Spectrometry

BACKGROUND:The umbilical cord mesenchymal stem cells possess multipotent differentiation capacity, but less research focus on its differentiation into fibroblasts.
OBJECTIVE:To investigate the capacity of human umbilical cord mesenchymal stem cells differentiating into fibroblasts.
METHODS:Using adherent method, human umbilical cord mesenchymal stem cells were isolated, and flow cytometric analysis of the surface antigen was performed. Passage 3 cells were selected for osteogenic and
adipogenic differentiation, and cells differentiated into fibroblasts under the induction of basic fibroblast growth factor. RESULTS AND CONCLUSION:Adherent stem cells were stably isolated from the umbilical cord. Human umbilical cord mesenchymal stem cells lowly expressed CD31, CD45, CD40, HLA-DR, but strongly expressed CD29, CD90, CD44, CD105. Oil red O staining showed red droplets were ful of the cytoplasm after adipogenic induction;alizarin red staining showed red calcium nodules after osteogenic induction. After induced by basic fibroblast growth factor, the type I col agen expression was significantly higher than that in the control group. These findings indicate that the adherent human umbilical cord mesenchymal stem cells are reliably isolated with high purity;basic fibroblast growth factor can induce differentiation of umbilical cord mesenchymal stem cells into fibroblasts.

Objective To assess the clinical usefulness and value of the 5 models for the prediction of acute kidney injury (AKI),severe AKI which renal replacement treatment was needed (RRT-AKI) and death after cardiac surgery procedures in Chinese patients.Methods One thousand and sixty-seven patients who underwent cardiac surgery procedures in the department of cardiac surgery in the Zhongshan Hospital,Fudan University between May 2010 and January 2011 were involved in this research.The predicting value for AKI (AKICS),RRT-AKI (Cleveland,SRI and Mehta score) and death (EURO score) after cardiac surgery procedures was evaluated by Hosmer-Lemeshow goodness-of-fit test for the calibration and area under receiver operation characteristic curve (AUROC)for the discrimination.Results The incidence of AKI was 20.34％(217/1067),and 63.13％ of their renal function recovered completely.The incidence of RRT-AKI was 3.56％(38/1067) and the mortality of AKI and RRT-AKI was 9.68％ (21/217) and 44.73％ (17/38) respectively.The total mortality was 3.28％ (35/1067).The discrimination and calibration for the prediction ofAKI of AKICS were low.For the prediction ofRRT-AKI,the discrimination and calibration of Cleveland score were high enough,but the predicated value was lower than the real value (1.70％ vs 3.86％).The discrimination of Mehta score and the calibration of SRI were low.The discrimination and calibration for the prediction of death of EURO score was low.Conclusion According to the 2012 KDIGO AKI definition,none of the 5 models above is good at predicting AKI after cardiac surgery procedures.Cleveland score has been validated to have a proper impact on predicting RRT-AKI after cardiac surgery procedures,but the predicting value is still in doubt.EURO score has been validated to have an inaccurate predicting value for death after cardiac surgery procedures.

Animals ; Carcinogens ; toxicity ; Female ; Lung Neoplasms ; chemically induced ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Sensitivity and Specificity ; Urethane ; toxicity

Objective To study the relative bioavailability after ig rats with Ophiopogon japonicus saponin enteric microsphere in single-dose so as to give directly preparation quality evaluation. Methods(Establishing the) analysis method of HPLC-MS and dealing with the blood sample by solid phase extraction. Determining the diosgenin concentration in plasma after ig Ophiopogon japonicas saponin enteric microsphere and the suspension as comparison in single-dose, and calculating pharmacokinetic parameters and relative bioavailability. Results C_(max), t_(max), and AUC_(0-72) were (637.81?17.23) ng/mL, (4.0?0.0) h, and 7 840.01?412.03, respectively. Conclusion The characteristics of absorption, distribution, and elimination are resembled in rats between two preparations by ig in single-dose. The pharmacokinetic parameters are also identical. The relative bioavailability is (91.10?3.44)%.

OBJECTIVETo screen miRNA profiles of malignantly transformed human bronchial epithelial cells, 16HBE-T, induced by anti-benzo[a]pyrene-trans-7,8-diol-9,10-epoxide (anti-BPDE), and to analyze putative miR-10a targets in 16HBE-T.

METHODSA novel microarray platform was employed to screen miRNA profiles of 16HBE-T cells transformed by anti-BPDE. Microarray data for miR-10a and miR-320 were validated using quantitative real time polymerase chain reaction (QRT-PCR). The expression of a putative target for miR-10a, HOXA1, was analyzed by reverse transcription polymerase chain reaction (RT-PCR) and QRT-PCR.

RESULTSIn comparison with the vehicle-treated cells (16HBE-N), 16HBE-T exhibited differential expression of 54 miRNAs, in which, 45 were over-expressed and 9 were down-regulated. The five most highly expressed miRNAs were miR-494, miR-320, miR-498, miR-129, and miR-106a. The lowest expressed miRNAs were miR-10a, miR-493-5p, and miR-363*. Three members of miR-17-92 cluster, miR-17-5p, miR-20a, and miR-92, showed significantly higher abundance in 16BHE-T as miR-21, miR-141, miR-27a, miR-27b, miR-16 and miRNAs of the let-7 family. The putative target for miR-10a, HOXA1 mRNA was up-regulated 3-9-fold in 16HBE-T, as compared with 16HBE-N.

CONCLUSIONThe findings of the study provide information on differentially expressed miRNA in malignant 16HBE-T, and also suggest a potential role of these miRNAs in cell transformation induced by anti-BPDE. HOXA1 is similarly up-regulated, suggesting that miR-10a is associated with the process of HOXA 1-mediated transformation.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide ; toxicity ; Carcinogens ; toxicity ; Cell Transformation, Neoplastic ; chemically induced ; genetics ; metabolism ; Cells, Cultured ; Gene Expression Profiling ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; MicroRNAs ; metabolism ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics ; metabolism

OBJECTIVETo screen microRNA (miRNA) profiles of malignantly transformed cells induced by anti-benzo-a-pyrene-trans-7, 8-dihydrodiol-9, 10-epoxide (BPDE) and to look for miRNAs which is expressed differently between malignantly transformed cells and normal human bronchial epithelial cells 16HBE.

METHODSExperimental group was the malignantly transformed 16HBE which was induced by cultured with final concentration 2.0 micromol/L of BPDE which was dissolved in dimethyl sulphoxide. The control group was 16HBE that was cultured with minimal essential medium including dimethyl sulphoxide. 327 miR-NAs were tested be-tween those two groups with miRNA microarray analysis. MiR-10a that was down expressed and miR-320 that was overexpressed were selected to be validated by miRNA specific quantitative real-time reverse transcriptase chain reaction (miR qRT-PCR).

RESULTS327 human miRNAs were tested with miRNA microarray analysis. 55 miRNAs were found expressing differently between those two groups and of which 46 were overexpressed and 9 were down expressed. Some data were validated by quantitative RT-PCR.

CONCLUSIONmiRNAs expressed significantly between malignantly transformed 16HBE and normal cells and this helps us look for unique miRNAs of malignantly transformed cells induced by BPDE, but there should have more sufficient evidences to prove their functions in malignant cells.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide ; adverse effects ; Bronchi ; cytology ; Cell Transformation, Neoplastic ; chemically induced ; genetics ; pathology ; Cells, Cultured ; Epithelial Cells ; drug effects ; pathology ; Gene Expression Profiling ; Humans ; MicroRNAs ; genetics