Acrylonitrile ; toxicity ; Animals ; Male ; Mice ; Mice, Inbred Strains ; Testis ; drug effects ; pathology

Objective To construct the eukaryotic expression vector of Notch1intracellular domain,p3XFLAG?CMV7?NICD1,so as to prepare for the further research and exploration of effect of Notch1 on promoting epithelial?mesenchymal transition of human lens epithelial cells. Methods The cDNA fragment was reversely transcribed by RT?PCR from total RNA extracted from the SRA01/04 cells and was encoded with the specific am?plification?targeted NICD1was obtained from the SRA01/04 cells,then the cDNA fragment was inserted into p3XFLAG?CMV7 to transcribe Esche?richia coli DH5α. And the recombinant plasmid was extracted after bacterial screening by LB plating medium and confirmed by the restriction endo?nuclease digestion and DNA sequencing. Results The target gene obtained had the same molecular size as predicted. It was indicated that recom?bined p3XFLAG?CMV7 plasmid contained correct recombinant human Notch1 sequences and p3XFLAG?CMV7?NICD1 was constructed success?fully. The western blotting showed protein NICD1 expressed in SRA01/04 cells transfected with p3XFLAG?CMV7?NICD1. Conclusion The suc?cessful construction of p3XFLAG?CMV7?NICD1 will provide a foundation for a further study studies in on the effect relationship of Notch signaling pathway and in posterior capsular opacification(PCO)after cataract extraction.

Objective To evaluate the relaxation rates and imaging effects of three MRI contrast agents gadopentetate dimeglumine(0.5 mol/L),gadodiamine and gadovist(1.0 mol/L) in the nervous system.Methods Relaxation rate differences between the three contrast agents were assessed using the GE Signa HDx 3.0 T MR scanner and phantom solutions of different albumin concentrations.Twenty leukemia patients whose initial scans had been conducted with the injection of a standard dose(0.5 mol/L)of gadopentetate dimeglumine as the contrast agent were recalled to have a follow-up scan for signs of brain infections with the same imaging protocols,except that a high concentration(1.0 mol/L)gadovist was used this time as the contrast agent.CNR and SNR in the ROI were measured for quantitative analysis.Results Changes in dosage of the three contrast agents produced no difference in intensity of the image signal for each phantom solution of a specific albumin concentration(5.0 g/L:P=0.35,6.5g/L:P =0.27,8.0 g/L:P=0.23).Two sets of scans of the leukemia patients showed that high concentration(1.0 mol/L)gadovist generated higher SNR and CNR in the ROI of the white matter,gray matter and vasculature than standard concentration(0.5 mol/L) gadopentetate dimeglumine(P＜ 0.05).Conclusions A half dose of high concentration(1.0 mol/L) gadovist generates better imaging enhancement than standard concentration(0.5 mol/L)gadopentetate dimegluminethe.Gadopentetate dimeglumine,gadodiamide and gadovist have no significant difference in relaxation rate.

Objective To explore the risk factors of acute organophosphorus pesticide poisoning(AOPP). Method The patients with acute organophosphorus pesticide poisoning,admitted from September 2004 to August 2005,were retrospectively analyzed.The 87 patients were divided into groups according to the presence or absence of combined conditions including hypotension,hypoxemia or metabolic acidosis.Acute physiology and chronic health evaluationⅡ(APACHEⅡ)score was calculated.The chi-square test was used to examine the mortality between those groups.Results The total in-hospital mortality of 87 acute organophosphorus pesticide poisoning patients was 21.8%.The mean APACHEⅡscore was(7.58?5.32)in the 68 survivors and(21.17～9.46)in the 19 dead,there were significant differences between the survivors and the dead(t=9.25,P20 was 65.2%(15/23),andit was 6.3% (4/64)in patients with APACHEII score

OBJECTIVE: To determinate triptolide and wilforlide A in biological samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method and to verify the method. METHODS: After 0.4 mL blood, urine or 0.4 g hepatic tissues with internal standard were extracted by ethyl acetate, they were separated on a Allure PFP Propyl (100 mm x 2.1 mm, 5 µm) with a mobile phase of methanol-20 mmol/L ammonium acetate using gradient elution. For mass spectrometric detection, electrospray ionization (ESI⁺) in positive mode was elected and the data was collected using multiple-reaction monitoring (MRM). RESULTS: The linearity was good (r > 0.995 0) and the limit of detection was 2 ng/mL or 2 ng/g for triptolide and wilforlide A. The recovery was 61.08%-102.98%. The intra-day and inter-day precision was less than 12.58% for each biological sample, and the accuracy was 90.61%-105.80%. CONCLUSION This method is simple, convenient and good selective, and could be applied to analysis of triptolide and wilforlide A in different biological samples. And the method may provide technical support for forensic medicine identification, clinical diagnosis and treatment of tripterygium wilfordii Hook. f. poisoning.
Chromatography, High Pressure Liquid ; Diterpenes/urine* ; Epoxy Compounds/urine* ; Humans ; Oleanolic Acid/urine* ; Phenanthrenes/urine* ; Sensitivity and Specificity ; Tandem Mass Spectrometry

Acute Disease ; Adolescent ; Adult ; Drugs, Chinese Herbal ; therapeutic use ; Ethylene Dichlorides ; poisoning ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Middle Aged ; Poisoning ; drug therapy ; Retrospective Studies ; Treatment Outcome ; Young Adult

Adoptive Transfer ; methods ; trends ; Cells ; pathology ; Cells, Cultured ; pathology ; Humans

Adenocarcinoma ; metabolism ; pathology ; surgery ; Aged ; Humans ; Intestinal Neoplasms ; metabolism ; pathology ; surgery ; Intestine, Small ; Ki-1 Antigen ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; surgery ; Male ; Neoplasms, Multiple Primary ; metabolism ; pathology ; surgery ; Rectal Neoplasms ; metabolism ; pathology ; surgery

The directed molecular evolution of enzyme in vitro can not only improve the efficiency of evolution, but also evolve enzyme according to the investigator's desire. This review summed up the feasible methods of this novel technique.

OsRacD belonging to rice Rho family of the small GTP binding proteins,is a pivotal gene involved in rice photoperiod fertility conversion of photoperiod sensitive genic male sterile rice,which influences rice fertility via controlling the pollen tube growth.T20N site-directed mutation was introduced into its highly conserved G1 motif by PCR-mediated method to mimic its GDP-binding state based on the sequences alignment and conserved domains analysis.The prokaryotic expression vector of OsRacD and T20N-OsRacD were constructed,and the His6 tag fused proteins were expressed and purified from E.coli.After identified by Western blot,the GTP hydrolysis activities were detected.The results showed that the GTPase activities of T20N-OsRacD were significantly reduced comparing with that of OsRacD,suggested that OsRacD and T20N-OsRacD have different biochemical characteristics.