The 6.5kb specific fragment containing the T7 promoter and the transcription vector was cut down from the full-length cDNA clone of Newcastle disease virus strain ZJI of goose origin,and thereafter it was self-ligated to form the high guality plasmid for mutagenesis.Site-directed mutagenesis technique was used for inserting three additional G nucleotides(nts) into the region between the T7 promter and the leader sequence of the NDV.The RT-PCR was employed to amplify the F/HN genes fragements,and then they were ligated by the shairing restriction enzyme BsmBI and finally the corresponding fragment in the mutatant full-length cDNA was substituted by the new one.The sequencing results showed that the three additional G nts were successfully inserted and the mutant nts in the full-length cDNA were corrected and all these studies lay a foundation for the research on the reverse genetics of NDV strain ZJI.

[Objective] To explore the characteristics and achievements of YU Chang'sYuyi Grassin treating typhoid disease. [Method] Based on studying, thinking and analysing theYuyi Grass, combined with related literature research, it summed up the YU Chang characteristics and achievements in treating typhoid disease. [Results] YU Chang treating typhoid fever syndrome had accuracy differentiation, forcefully rectifying children with typhoid fever being mistaken for the fallacies of present concern of charlotte, fine solution of typhoid rehabilitation treatment; slow treatment for typhoid critical patients, illustrating the appropriate use of ginseng theory for typhoid fever, having the original opinion in such use. [Conclusion] The Yuyi Grass treating typhoid disease has rich experience and unique achievements. These insights and achievement in learning by using the theory of Treatise on Febrile Disease for treating typhoid disease have important guiding significance in Chinese medicine clinic.

Objective To study the related factors of operating on Pilon bone fracture.Methods 35 cases with tibial Pilon fracture treated operative,according to the Reedi-Allgower were divided the type,Ⅱ type 15 examples,Ⅲ type 20 example.14 cases were treated with open redcution and inter fixation plate and screws and 11 cases were treated with open reduced minimal and exter fixation that cast.Results According to Mazur's grade system treatment was evaluated may in 28 cases,18 cases of which were type Ⅱ racture and 10 cases type Ⅲ fracture poor in 7 cases,2 cases of which were type Ⅱ fracture and 2 cases type Ⅲ fracture.Conclusion The curative effect with fracture type,repla-cement mass and operation time are relative in tibial Pilon fracture.

As an energy dependent membrane transporter, P-GP highly expresses in the organs which have the function of barrier and secretory.In the brain, P-GP mainly expresses in the cerebral vascular endothelial cells of blood brain barrier.In the state of brain injury diseases ( such as epilepsy and cerebral ischemia) , P-GP appears in the hippocampus, cortex and striatum of rat brain.At cellular level, it mainly exists in the cerebral vascular endo-thelial cells and also overexpresses in the neurons and glial cells.

Objective To investigate the employment pressure among graduates from nursing school,and explore influencing factors,in order to make suggestions on education management,career guidance and psychological intervention.Methods Students graduated from Nanyang Medical College in 2012 who majored in nursing and nursing graduates from Nanyang Institute of Technology were surveyed with medical students employment pressure questionnaire designed by DU Tian-jiao from China Medical University.SPSS13.0 was used to establish database and analyze data.Results The total employment pressure of all students was M=3.2.Seven stressors scores from high to level were:post requirements,social environment,family support,personal qualities,the school effect,professional requirements,and career guidance.Multiple regression analysis showed that whether or not the student leaders,different sources and the education background were the influencing factors of employment pressure.Conclusions This research suggests that different psychological intervention should be given to students majored in different specialty and sources in order to release employment pressure and anxiety.

Objective To explore the clinical guiding value of spiral computed tomography(SCT)with three-dimensional computed tomography reconstruction(3-DCT)in percutaneous kyphoplasty(PKP).Methods 24 patients prepared to be treated with PKP were examined under 3-DCT before the treatment.Results According to the contra-indications shown by 3-DCT,11 patients' operation were cancelled;the others were treated with PKP and got good outcomes due to the accurate data provided by 3-DCT.Conclusion SCT plays an important role in PKP.

Botulinum neurotoxin(BoNT) is the most lethal biotoxin known to mankind. It inhibits acetylcholinerelease from the cholinergic nerve ending by cleavage of SNARE proteins, followed by neuromuscular blockadeand paralysis. Gangliosides are considered to act as a first receptor of BoNT with low affinity.Then the membranebound gangliosides-BoNT complex moves laterally to reach and bind the toxin specific protein receptor,synaptotagmin, with a high affinity constant. At last the gangliosides-BoNT-synaptotagmin complex undergoesreceptor-mediated endocytosis. This double-receptors theory is widely accepted. The research data are summarizedand reviewed.

Usage of the fruit and bark of a Melia-family plant as a digestive tract-parasiticide and agricultural insecticide was recorded about two thousant years ago in ancient China. Toosendanin (TSN), a triterpenoid, is an effectual ingredient extracted from the plant. Studies have demonstrated that TSN selectively affects neurotransmitter release, effectively antagonizes botulism, induces cell differentiation and apoptosis and inhibits proliferation of various human cancer cells, inhibits feeding and dovelopment in insects and modifies K+- and Ca2+-channel activity. The research data to demonstrate that TSN inhibits K+-channel and facilitates L-type Ca2+-channel are summarized, and the mechanism of action of TSN is discussed.

Objective To investigate the expression levels and significance of interferon-γ(IFN-γ),interleukin (IL)-10 mRNA in peritoneal fluid mononuclear cells (PFMC) in patients with endometriosis (EM).Methods A total of 55 patients with EM diagnosed histopathologically after operation were enrolled as EM group,and 52 patients without EM were enrolled as control group.The PFMC were isolated,and the expression levels of IFN-γ and IL-10 mRNA were determined by TaqMan real-time reverse transcriptase-polymerase chain reaction (RT-PCR).Results It was found that IFN-γ mRNA expression level was significantly decreased in EM group compared with that in control group (P＜0.05),while IL-10 mRNA expression level was significantly increased in EM group compared with that in control group (P＜0.05).The ratio of IFN-γ/ IL-10 was significantly lower in EM group than that in control group (P＜0.05).The ratio of IFN-γIL-10 was significantly higher in patients with stage Ⅰ-Ⅱ than that in patients with stage Ⅲ-Ⅳ (P＜0.05).Conclusion The imbalance of IFN-γ/ IL-10 in PFMC is associated with the pathogenesis of EM.

Objective:To study the chemical constituents in the root of Petasites tricholobus Franch.and evaluate their anti-inflammatory activity.Methods: The primary extraction was done with 95% ethanol and subsequently with other agents including ethyl acetate.The compounds extracted with ethyl acetate were isolated by silica gel column chromatography and purified by means of recrystallization.The structures of the compounds were elucidated by employing chemical and spectral methods.The anti-inflammatory activities of the compounds were evaluated through their inhibitory effect on the trachea contraction of guinea pig induced by histamine in vitro.Results: Six compounds were isolated from the root of Petasites tricholobus Franch.,namely,homofukinolide(1),bakkenolide-B(2),stigmasterol(3),and ?-sitosterol(4),bakkenolide-D(5),and ?-sitosterol-3-O-?-D-glucopyranoside(6).Bakkenolide-B and bakkenolide-D demonstrated inhibitory effect on the trachea contraction induced by histamine in vitro.Conclusion: Homofukinolide,?-sitosterol,and ?-sitosterol3-O-?-D-glucopyranoside are isolated for the first time from Petasites tricholobus Franch..Bakkenolide-B and-D have obvious anti-histamine activities.