Omnibearing reform and practice of medical education based-on integrated organ system-based curriculum and PBL / 中华医学教育探索杂志

Xi'an Jiaotong University has proposed the concept of "less teaching and more learning, interaction between guiding and learning" in medical education, based on its sedimentary deposits, and carried out reform for all clinical medical students since 2001. After more than ten years of educational reform, we have built brand new management framework, and established integrated organ system-based curriculum and PBL teaching pattern. This pattern involves eight aspects of comprehensive reform, including training program, curriculum model, textbook, teaching method, learning style, assessment and evaluation, teaching organization, teaching conditions and guarantee. It will provide paradigm for the integrated curriculum reform in peer colleges, and will be a milestone in the history of medical education in China.

Construction and verification of anti-MM scFv-tP fusion protein expression vector / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct an expression vector of anti-MM scFv-tP fusion protein and test its expression efficiency and function.</p><p><b>METHODS</b>The truncated protamine (tP) gene sequence was added to the gene of single chain antibody against the specific antigen on the surface of malignant melanoma tumor cells using PCR. A GST-fusion expression vector was constructed and the soluable protein was expressed in the E.coli system. After cleavage and purification, the purified fusion protein was obtained. The binding activity of Anti-MM scFv-tP and siRNA was detected by EMSA. Flow cytometry and confocal microscopy were used to detect the cell surface antigen binding activity of the fusion protein.</p><p><b>RESULTS</b>The expression vector of Anti-MM scFv-tP fusion protein was successfully constructed. The soluable protein could be expressed in the E.coli system, and the purified fusion protein was obtained. The anti-MM scFv-tP fusion protein retained siRNA binding ability and could directly target malignant melanoma (MM) LiBr cells.</p><p><b>CONCLUSION</b>The recombinant GST- Anti-MM-scFv-tp expression vector was successfully constructed. The fusion protein retains siRNA binding ability and can directly target LiBr cells to provide a reliable tool for further study.</p>

Effects of spironolactone towards rabbit atrial remodeling with rapid pacing

This study aimed to observe the effects of spironolactone towards the rabbit atrial remodeling with rapid atrial pacing [RAP]. 30 rabbits were randomly divided into control group, RAP group and spironolactone group, with 10 rabbits in each group. RAP was performed at the speed of 800 beats/min for 8 h, atrial effective refractory period [AERP] was determined before and at the 1[st], 2[nd], 4[th], 6[th] and 8[th] of the pacing, the expressions of atrial muscular calcium channel alpha 1C subunit and beta 1 subunit mRNA were performed the RT-PCR detection, and ultrastructural changes of atrial myocytes were observed. AERP of RAP group shortened, with poor frequency adaptability; the expressions of calcium channel alpha 1C subunit and beta 1 subunit mRNA decreased 22% and 26%, respectively, when compared with the control group; ultrastructure of atrial myocytes changed significantly. AERP of spironotlactone group shortened less that RAP group, and the frequency adaptability was maintained, the decreased expressions of calcium channel alpha 1C subunit and beta 1 subunit mRNA significantly reduced. RAP could cause atrial remodeling, while spironolactone could inhibit RAPinduced atrial remodeling

Quantitative determination of metoprolol by liquid chromatography mass spectrometry in rat plasma / 军事医学科学院院刊

Objective:To establish a highly sensitive, rapid and selective liquid chromatography mass spectrometry (LC-MS) method for the determination of metoprolol in rat plasma.Methods:A simplified liquid-liquid extraction with acetidin was employed for the sample preparation. The separation was carried out on a Thermo ODS-C_(18)(5 μm,100 mm×2.1 mm).The mobile phase consisted of acetonitrile-methanol-water(20∶20∶60). Propranolol was used as the internal standard. The detection was performed on a liquid chromatography mass spectrometry by selected ion monitoring(SIM) scan mode electrospray ionization(ESI).Results and Conclusion:The range of calibration curve was 0.1-50 ng/ml and the limit of quantification was 0.1 ng/ml. The intra- and inter-day precision RSD was less than 15%.This method is sensitive, simple,rapid and suitable for the pharmacokinetic study of metoprolol.

Pharmacokinetics of 6beta-naltrexol after single and multiple intramuscular injections in Beagle dogs / 药学学报

The pharmacokinetics of 6beta-naltrexol (6beta-NOL) following single intramuscular administration and multiple intramuscular injection once per day for seven days was studied in 4 Beagle dogs. Plasma concentration of 6beta-NOL in dogs was analyzed by a combination of high performance liquid chromatography (HPLC) and electrochemical detection with naloxone (NLX) as internal standard. After single intramuscular injection of 0.2 mg x kg(-1) 6beta-NOL, the plasma concentration-time curve of the drug was found to fit to a two compartment model with first-order absorption. The main parameters of single dosing were as follows: t1/2alpha was (0.26 +/- 0.23) h, t1/2beta was (4.77 +/- 1.65) h, C(max) was (81.65 +/- 5.61) ng x mL(-1), t(peak) was (0.27 +/- 0.07) h, CL(s) was (1.20 +/- 0.06) L x kg(-1) x h(-1), V/F(c) was (1.94 +/- 0.15) L x kg(-1), and AUC(0-t) was (166.82 +/- 7.68) ng x h x mL(-1), separately. After multiple intramuscular injection of 0.2 mg x kg(-1) 6beta-NOL once per day for seven days, the plasma concentration-time curve of the drug fitted to a two compartment model with first-order absorption too. The main parameters of the last dosing were as follows: t1/2alpha was (0.19 +/- 0.18) h, t1/2beta was (5.79 +/- 1.50) h, C(max) was (79.82 +/- 10.5) ng x mL(-1), t(peak) was (0.18 +/- 0.08) h, CL(s) was (1.12 +/- 0.07) L x kg(-1) x h(-1), V/F(c) was (2.10 +/- 0.27) L x kg(-1), and AUC(0-t) was (173.23 +/- 9.49) ng x h x mL(-1), separately. The difference of the parameters between the first and the last dosing was not significant, showing that the plasma kinetics of 6beta-naltrexol was not changed after multiple administrations. In the course of multiple administration, the peak and valley concentration of plasma 6beta-naltrexol were (79.03 +/- 10.3) and (1.50 +/- 0.93) ng x mL(-1), respectively. No clear adverse events were noted during this study. These results showed that plasma 6beta-naltrexol fits to a two compartment model with first-order absorption in dog after intramuscular administration and their pharmacokinetic parameters were reported. There was no remarkable change on plasma pharmacokinetics of 6beta-naltrexol after multiple intramuscular administrations.

Differential expression of genes related to transcription in cultured hepatoma cells with intact genome of hepatitis C virus / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of hepatitis C virus (HCV) on transcription regulation genes of host cells by gene chip assays in cultured cells with intact HCV genome.</p><p><b>METHODS</b>Huh-7 hepatoma cells were cultured and infected with in vitro constructed HCV. The total RNAs, proteins and cell culture supernatants of HCV infected cells and control cells were isolated. Proteins and cell culture supernatants were used to detect the HCV replication and protein expression in cell culture system. The HCV protein expression was detected with Western blotting. Released HCV from infected cells was analyzed by real-time fluorescence quantitative PCR. Total RNA was qualified using 10 g/L agarose gel electrophoresis. cRNA was synthesized, fluorescence labeled and purified, then hybridized with Agilent oligo microarray (20173 probes). Differential expression of genes related to transcription in cell culture system was analyzed.</p><p><b>RESULT</b>HCV was positive in cell culture supernatants and HCV protein expression was also positive according to Western blotting results. Eleven up-regulated and 11 down-regulated genes related to transcription were found after Agilent gene chip screening.</p><p><b>CONCLUSION</b>Intact hepatitis C virus cell culture system provides an useful tool for study on the affects of HCV infection on transcription regulation genes in host cells.</p>

Current status and perspective of hepatitis C virus models / 浙江大学学报·医学版

Lack of proper study models has brought difficulties in the study of the mechanism of viral infection, life cycle and pathogenic mechanism of hepatitis C virus (HCV) and also become the major obstacles in development of efficient vaccine and new drugs for hepatitis C. In recent years, the establishment of robust HCV cell culture infection system and HCV transgenic animal provide powerful tools for the analysis of host virus interactions, which facilitate the discovery of antiviral drugs and vaccines for this important human pathogen.

Analysis of ORF27 full length sequence of Thai isolated Helicoverpa armigera nucleopolyhedrovirus and prokaryotic expression of truncated ORF27 gene / 西安交通大学学报(医学版)

Objective To understand the functions of ORF27 protein of the Thai isolated Helicoverpa armigera nucleopolyhedrovirus(HaNPV-Th) and explain the possible molecular mechanism of HaNPV-Th in passage effect.Methods ORF27 gene was cloned and amplified by polymerase chain reaction(PCR) from HaNPV-Th genome.The physical-chemical properties,subcellular localization,signal peptide,transmembrane structure and functional motifs were analyzed by the software of ProtParam,TMpred,SignalP and ScanProsite.The truncated ORF27 gene which lacked parts of the amino-terminal was expressed by bacterial expression system.Results The full length sequence of ORF27 was 768bp long and encoded 255aa with a molecular weight of 29.5ku.Several functional motifs on ORF27 were predicted by bioinformatics software.The truncated ORF27 recombinant expressed 35ku target protein mainly in the form of inclusion body.Conclusion ORF27 protein of HaNPV-Th is presumed to possess extensive biological activities.It may regulate various cellular signaling pathways,apoptosis and cell cycle,which are involved in passage effect.