Objective To construct human acidic fibroblast growth factor (aFGF) recombinant eu karyotic expression vector and transfect it into muscle satellite cells(MSCs) of rat,in purpose of further study the method to set up cell bank.Methods The aFGF gene was cloned from human total RNA which was obtained from human skeletal muscle tissue by RT-PCR method.Human interleukin 2 (IL-2) signal peptide sequence (SPS) was obtained by direct chemosynthesis method.Then aFGF and SPS were fused to obtain SPS-aFGF.Finally,directional cloning SPS-aFGF into pEGFP-N1,the recombinant (pEGFP-N1-SPS-aFGF) was obtained.The recombinant was confirmed by endonuclease digestion and DNA sequencing.MSCs were purified by difference-speed adherence method and were ideontified by immunofluorescence assay.The correct cells were divided into 3 groups:Experimental group (aFGF +N 1),control group (N 1),blank group (blank).All the groups were transfected by Lipofectamine 2000TM Reagent,and pEGFP-N1-SPS-aFGF,pEGFP-N1 were respectively added in experimental group and control group while blank group was added none plasmid.Fluorescence microscope was employed to detect transfection efficiency tendency along with time changes.The expression of target gene was detected by fluorescent quantitation PCR and Western blot.Results (1) The sequencing of pEGFP-N1-SPS-aFGF was completely correct and the outcome of endonuclease was equal to actual ban s-ize.(2)The expression of GFP in transfected cells were observed by fluorescencemicroscope and transfection efficiency reached the peak at 72 h.(3)Real-time fluorescent quantitation PCR proved strong aFGF mRNA expression in transfected cells (the average relative expression of experimental group was 1464.95)with aFGF gene,while it was detected a little in the other groups (the average relative expression of control group was 1.016 and blank group was 1.000) (P ＜ 0.05).Western blot also proved strong expression in Experimental group then the other two groups.Conclusion aFGF eukaryotic expression vector was successfully constructed and transfected into MSCs.This study may be expected to obtain some specific functions cells.

Objective To evaluate the performance of ACL TOP coagulation analyzer system in the laboratory of children′s hospital. Methods According to the documents of CLSI, the analytic characteristics including precision, accuracy, linearity, interference and carryover rate were examined; specimens from healthy children were collected and assayed to determine the reference range of Prothrombin Time (PT), Activated Partial Thromboplastin Time (APTT), Thrombin Time (TT), Fibrinogen (FIB) and D-Dimer (D-D) from children on ACL TOP. Results The within-run and between-day coefficient of variability (CV) were within an acceptable range; The accuracy deviation of PT , APTT and FIB were less than 1/2 allowed total errors; The results of determination of FIB linearity test were correlated with the results of calculation: Y = 1.002 1X-0.122, R2 =0.998 2; The extent of influence of low to middle grade of jaundice , fat and hemolysis on each test were all less than 1/2 allowed total error; The carryover rates were lower than 1.81% and within an acceptable range; The reference range of PT, APTT, TT and FIB were PT (9.1-13.1 s), APTT (24.9-42.1 s), TT (12.6-21.1 s), FIB (1.924-4.011 g/L). Conclusion The ACL TOP coagulation analyzer has good repeatability, stability, linearity and capability of anti-interference and anti-carryover.

Objective To investigate the knowledge of chronic heart failure (CHF) and influencing factors in general practitioners (GP) in Beijing.Methods A self-designed questionnaire contained total 28 items,including basic knowledge of CHF,non-drug management,drug management and other management ; clinical cases were used to test clinical ability in 7 items.The questionnaire survey was conducted among GPs who participated in continuing education courses from 16 counties/districts in Beijing during January to June 2013.Results Total 720 questionnaires were distributed and 657 valid questionnaires were returned with a recovery rate of 91.3％.Total scores was 60.6.Scores of basic knowledge,non-drug management,drug management and other management was 63.1,76.9,44.0 and 56.9,respectively.There were statistical differences in scores of basic knowledge,non-drug management,drug management and other management between GPs with different diploma (F value:36.8,5.8,21.6,12.2,respectively; P ＜0.01) ; there were significant differences in scores of basic knowledge and drug management among GPs with different working years (F value:15.1 and 17.4,respectively ; P ＜ 0.01) ; there was significant difference in scores of drug management among GPs with different professional title (F =7.69,P ＜ 0.01).Only for GPs with junior college diploma,the scores of basic knowledge and drug management in GPs with working ≥20 y were higher than those working ＜ 20 y(P ＜ 0.01).The accuracy of clinical ability in GPs with junior college diploma,undergraduate diploma and post-undergraduate diploma was 39.6％,41.6％,41.8％ (P ＞ 0.05).Conclusions The knowledge of CHF is less desirable in GPs of Beijing,so that measures should be taken to improve the GP's knowledge of CHF.

Brain ; diagnostic imaging ; Child, Preschool ; Electroencephalography ; Female ; Gastroenteritis ; complications ; Humans ; Infant ; Male ; Seizures ; etiology ; Tomography, X-Ray Computed

Objective: To study the UPLC-MS fingerprint of Bupleurum marginatum DC. in northwest Hubei and establish the quality evaluation system for the herb. Methods:A UPLC-MS method was used to analyze 10 samples of B. marginatum DC with the following chromatographic conditions:an ACQUITY UPLC? RBEH C18 column (100 mm × 2. 1 mm, 1. 7 μm) was used, the mobile phase consisted of acetonitrile-0. 3% formic acid water with gradient elution, the flow rate was 0. 2 ml·min-1, and the detection wavelength was 203nm with ESI(-). Results:There were 8 common peaks in the UPLC-MS fingerprint of B. marginatum DC. Through analyzing the information of mass spectrometry and combining the references, 6 peaks were identified. Conclusion:The UPLC-MS fin-gerprint method is simple,rapid and feasible. The acquired UPLC-MS fingerprint of B. marginatum DC. and the evaluation indices can provide the scientific quality assessment of B. marginatum DC.

Objective To study the effects of mechanical strain on the proliferation of the human pulmonary epithelial cell and the redistribution of its membrane receptors, integrins ? 5 and ? 1. Methods A cyclic strain unit in vitro was designed. The cellular proliferative index was measured by flow cytometry and the redistribution of ? 5 and ? 1 integrins was analyzed in human pulmonary epithelial cell line H727 by laser confocal microscopy. Results The cellular proliferative index reduced significantly after cells were subjected to 15% elongation at frequencies of 20 cycles/min or 40 cycles/min for 24 h. In human pulmonary epithelial H727 cells, ? 5 and ? 1 integrins transferred from the apical layer to the basal layer and formed an adhesion plaque after 24 h exposure to 15% elongation at frequency of 40 cycles/min. Conclusion The results suggest that ? 5 and ? 1 integrins in pulmonary epithelial cells may play an important role in the transduction of mechanical stress.

Objective To determine the aromatase protein expression in A549 cell and to investigate the effects of 17? estrogen (E 2), testosterone (T), estrogen receptor antagonist tamoxifen (TAM), and aromatase inhibitor DL aminoglutethimide (AMIN) on the growth and proliferation of A549 cells. Methods The expression of aromatase protein was determined by immunohistochemical methods. The changes of cell cycle and cell number before and after treatment with E 2, T, TAM, and AMIN were measured by flow cytometry and tetrazolium method (MTT). Results The aromatase protein was positively expressed in A549 cells. The aromatase inhibitor AMIN and 5?10 -7 mol/L TAM could inhibit the growth of A549 cells and block them in G 0/G 1 phase ( P

Objective To use inverse distance weighting(IDW)in studying the distribution of endemic fluor.0sis in Jiangsu Province and evaluate the value of IDW in endemic fluorosis surveillance.Methods A geographic information system(GIS)database of endemic fluorosis was established in Jiangsu Province from the data of endemic fluorosis survevs conducted during 1982-1985.With the help of Arc View 3.3 system,IDW was applied to forecast the distribution of fluoride concentration in water and the distribution of the prevalence rate of dental fluorosis in Jiangsu Province based on the electronic map of Jiangsu Province.Results IDW was applied to forecast the distribution of endemic fluorosis in Jiangsu Province.By comparing with the result of endemic investigation in the 1980's.the forecasting Was proven to be accurate,exact and detailed.Conclusion With the application of IDW and stratified sampling,it is feasible to describe the spatial distribution of endemic fluorosis in Jiangsu Province in endemic fluorosis surveillance.

PAK1 plays an important role in the development of tumors. It is of great significance to screen and develop new PAK1 inhibitors as targeted drugs for cancer treatment. The traditional PAK1 inhibitor screening method has the problems of high cost and low efficiency. Computer virtual screening can reduce the cost of finding active lead compounds and improve the screening efficiency. In this study, a kind of PAK1 candidate compound was screened by computer assisted virtual screening combined with Z′lyteTM high flux kinase screen. In vitro enzyme activity screening showed that compound 18(K788)had good PAK1 inhibitory activity(inhibition rate was 42. 7%). Furtherly by MTT detection, it was found that K788 had significant PAK1 positive tumor killing activity, which was even better than the positive drug IPA-3. Flow cytometry and Western Blot showed that K788 could activate caspase apoptosis pathway and induce apoptosis of colon cancer cell DLD-1 by inhibiting PAK1 expression and activation. K788 has great potential for clinical development and application, and can be used as a PAK1 target for further research.

This research is to establish an HPLC method for the simultaneous determination of ophiopogonin D, ophiopogonin D', ophiopogonin C, deacetylophiopojaponin A and ophiogenin-3-O-α-L-rhamnosyl-(1-->2)-β-D-glucoside in Ophiopogonis Radix. HPLC-ELSD analysis was performed on a Kromasil 100-5 C₁₈ column (4.6 mm x 250 mm, 5 µm), with the mobile phase of acetonitrile (A) -water (B) in gradient elution mode (0-45 min, 35%-55% A), at a flow rate of 1 mL · min⁻¹. The column temperature was 35 °C and the drift tube temperature was 100 °C in a gas flow rate of 3.0 L · min⁻¹. The result showed that baseline of all the 5 constituents was well separated, and every constituent had wide linearity range and good linear relation (r > 0.999). The recovery rate was between 95.75% and 103.1%. The new established method for simultaneous determination of saponin constituents in Ophiopogonis Radix was sensitive and has good, repeatability. It could be applied to quality evaluation of Ophiopogonis Radix.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ophiopogon ; chemistry ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification