Objective To describe the anxiety status of new nurses in emergency department so as to put forward strategies for nursing management. Methods Thirty-two new nurses (working time less than 2 years) from emergency department of two class Ⅲ- A general hospitals in Xiamen were surveyed by using Zung self-rating anxiety scale (SAS). Results The average of SAS total score showed significant difference from that of the Chinese norm, but no significant difference could be seen in the score of SAS be-tween college and baccalaureate nurses.Nurses with different working length showed significant difference in SAS score. Conclusions Anxiety status of new nurses in emergency department is serious, nursing man-agers should make more efforts to conduct mental health education and improve psychological quality of new nurses in emergency department to relieve their anxiety.

Objective To establish the sperm specific Sleeping Beauty ( SB ) transposase-expression transgenic mouse for the study of the genetic modification mediated by transposon system in mouse .Methods Prm1 promoter was cloned from mouse genomic DNA to drive the expression of SB transposase .The transgenic mice were generated by microinjection .The gene type of transgenic line was identified by PCR .The expressing level in testis was determined by western blot and immunohistochemistry (IHC) staining.Results Five lines of transposase transgenic mice were obtained by microinjection and three can be germline .One mouse line with higher expression level of transposase in the testis was obtained.Conclusion One transgenic mouse model with Sleeping Beauty transposase - expression was successfully established .This model will greatly contribute to the research of genetic modification mediated by transposon in mouse.

Objective To develop a model that could roundly show the phenotypes of human alzheimer disease (AD), the triple-transgenic rat model harboring APP(Swe), PS1dE9, and TAU transgenes was established in view of the advantage of rat as an important animal model on the research of nerve system .Methods APPswe/PS1dE9/TAU triple transgenic rat AD rats were generated on a SD background by co-injecting rat pronuclei with two human genes driven by the mouse prion promoter:‘Swedish’ mutant human APP (APPsw) and exon 9 mutant human presenilin-1 (PS1dE9) and human microtubule-associated protein tau gene under the control of PDGF promoter .Transgene integration was confirmed by genotyping and expression levels were evaluated by western blot ( WB ) of brain homogenates .The pathological changes were detected by human Abeta, TAU and Phospho-PHF-TAU immunohistochemistry staining (IHC).The behavioral and cognitive changes were evaluated by Morris water maze .Results One transgenic rat lines with high human APP ( Swe ) , PS1dE9, and TAU transgenic expression was selected from three transgenic founders .Compared with the wild type rat , the transgenic rat showed significant learning and memory impairments in the Morris water maze at 6 months of age .The triple transgenic rat manifested hyperphosphorylated tau and obvious aggregation of amyloid -β( Aβ) in the brain cortex and hippocampus.Conclusion APPswe/PS1dE9/TAU triple transgenic rat AD model was established .The triple transgenic AD rat fills a critical need for a next-generation animal model to enable basic and translational AD research .

Objective To investigate the clinical implications of portal vein bloodletting immediately before reperfusion during orthotopic liver transplantation(OLT).Methods Thirty-two patients with end-stage liver diseases undergoing non veno-venous OLT were divided into bloodletting group (n=21)and control group(n=11).During anhepatic phase,we maintained mean arterial pressure ＞70 mm Hg,cardiac index＞2.5 L·min-1·m-2 by infusion,norepinephrine and dopamine.Blood samples were taken at the time when portal vein was clamped(T1),the time when portal vein was unclamped (T2),10 minutes after neohepatic phase(T3),neohepatic phase 30 minutes(T4)for electrolytes,blood gas and plasma inflammatory cytokines.Hemodynamic and ventilation parameters were also recorded.Results There was no significant difference in mortality(X2=1.12,P＞0.05)and arrhythmia incidence (X2=1.73,P＞0.05)between the two groups.Serum calcium,magnesium were both significantly lower than normal.After anhepatic phase,potassium,tumor necrosis factor alpha,interleukin-6 in radial artery didn't alter significantly;Bloodletting had no effect on lactic acid.There was no significant difference in hemodynamic and ventilation parameters among four time periods.Conclusion Bloodletting seemed to have no effect on changes of internal environment.

Objective To knock out the Abcb1 gene of rat,and establish the Abcb1 humanized rat model based on the Abcb1 knock out rat.Methods The animal model was established using BAC and CRISPR/Cas9 technology,and was analyzed by PCR, RT-PCR and real-time PCR.Results Establishing a rat model expressing human Abcb1 stably by transfer the 153 kb BAC containing human Abcb1 promoter and cDNA into rat genome, and establishing the Abcb1 knock out rat at the same time.Establishing the Abcb1 humanized model by crossing these two strains together.The expression pattern of Abcb1 in Abcb1 humanized rat is different from the wild type rat.The Abcb1 humanized model express not only the human Abcb1 gene but also has similar expression pattern as human.Conclusions The Abcb1 knock out rat and the Abcb1 humanized rat were successfully established, and this model is close to human concerning about the drug metabolism related to Abcb1.

Objective To obtain more physiological data of Leptin knockout SD rats available for the user , the long-term observation of fasting blood glucose and pathological phenotypes were performed .Methods The protein expression levels in liver tissues were determined by western blot .Body weight of Leptin knockout rats ( Leptin-/-) and littermate lean rats (Leptin+/+) were weighed up at 1,3,6,8 months of age.Fasting blood glucose of Leptin +/+rats and Leptin-/-rats at 1,3,6,8 months of age were measured using One Touch? brand blood glucose monitoring systems.Pathological changes of pancreas and livers of Leptin -/-rats were observed by the method of HE staining and Immunohistochemistry (IHC).Results Short null Lepin proteins were expressed in liver tissues from Leptin -/-rats. Leptin-/-rats become heavier than Leptin +/+rats since they were one month old .The body weight of Leptin -/-rats at 8 months of age was twice as heavy as Leptin +/+rats, female Leptin-/-rats weighing 884g, and male Leptin-/-rats weighing 1200g.Overt hyperglycemia was observed during the first month after birth .Compared with Leptin+/+female rats,the fasting blood glucose of Leptin -/-female rats was increased by 40%-26%from 1 to 6 months old. After that, blood glucose values decreased and eventually become nearly normal at 8 months of age.Pathological examination indicated that Leptin -/-rats at 8 months of age had a fatty liver , more pancreas islets with lager volume and more beta cells with increased insulin secretion .Conclusion Leptin-/-rat were characterized by obesity , fatty liver, islet cell hyperplasia and early hyperglycemia .

Objective To study the relationship of insulin receptor substrate-1 (Irs1) and metabolic disease, we generated Irs1 gene knockout rat by CRISPR/Cas9 system.Methods Two sgRNA targeting sites were designed for Irs1 targeting.The Cas9 and sgRNAs were transcribed by T7 RNA polymerase in vitro.Cas9 mRNA and sgRNA mixtures were pooled and microinjected into one-cell fertilized eggs of SD rats to generate rats with targeted mutation .Results Five rats with the mutations were detected with the efficiency of 83%.Conclusion The Irs1 gene knockout rats generated in this study can be transmitted by germline .

Objective To investigate the functional role of 25 microRNAs in breast cancer ,and to find new tumor suppressor microRNAs that may serve as specific targets of new gene therapies . Methods Twenty-five microRNAs expression vectors were constructed and stably transfected into mouse mammary tumor cells 4To7 by Lipofectamine2000. Cells were selected with G418 and sorted by Flow cytometry.The cells in logarithmic phase were collected and 2 ×105 cells/mouse was inoculated into BALB/c mice via tail vein .Lungs were harvested 14 days after tumor cell inoculation , and the number of metastasis foci was counted .Results Mice inoculated with mir-449a-expressing 4To7 cells via tail vein developed reduced lung metastases compared with mice inoculated with negative control cells .Mice inoculated with mir-1935-expressing 4To7 cells via tail vein developed increased lung metastases compared with mice inoculated with negative control cells .Other twenty-three microRNAs neither promoted nor inhibited lung metastases of breast cancer .Conclusions Two of twenty-five microRNA were identified to be associated with breast cancer metastasis .MiR-449a may play a tumour suppressor role in the regulation of migration and metastasis in breast cancer .miR-1935 transgenic over-expression promoted tumor growth and metastasis .

Objective To study the effects of NRG2 on cardiac structure and function , we established the cardiac-specific human NRG2 transgenic mice and investigate the effect of NRG2 on cardiac structure and function under pressure overload situation .Methods The transgenic vector was constructed by insertion of the human NRG2 gene under the α-MHC promoter.The transgenic mice were generated by microinjection and were all maintained on a C57BL/6J genetic background .The genotype of transgenic mice was identified by PCR and the expression level of target gene was determined by western blot .Transverse aortic constriction ( TAC) was applied to prepare the pressure overload induced cardiomyopathy mice model .The cardiac structure and function of the transgenic mice were compared and analysized by echocardiographic and pathological observation .Results Transgenic mice with high level of NRG2 in heart tissues were established.The left ventricular wall thickness (LVPWD) was increased, and to 15.6% at 3 months old compared with that of the non transgenic ( NTG) mice.The hypertrophy of left ventricular wall caused by pressure overload was removed due to the expression of NRG2 .Meanwhile, cardiac disarray and fibrosis were increased obviously compared with that of the NTG mice.Conclusion The transgenic expression of NRG2 in heart tissues could shorten the pathological process of hypertrophy, but accelerated the process of heart failure (HF).

Objective To explore differentiation in vitro of rat adipose-derived stem cells into photoreceptor cells and RPE cells.Methods The ADSCs were cultured by adhering to the flask surface and purified by continual passaging.Surface antigens including CD45、CD90、CD49d、CD106 were indentified by flow cytometry.ADSCs were induced to differentiate by EGF,activin A,taurine,retinoic acid(RA) and extracted liquid of retina respectively.Meanwhile,ADSCs were induced by EGF+taurine,EGF+RA,taurine + RA,EGF+taurine+RA respectively.Immunofluorescence was used for detecting the expression of rhodopsin,CK and S-100,and flow cytometry was used for quantification.Results For primary culture,the phenotypes of ADSCs were: CD45,CD90,CD49d and CD106,with a positive percentage of 1.6%,71.3%,7.8% and 3.5%,respectively.From passage 1 to 5,these phenotypes were: CD45(0.8%～9.3%),CD90(84.7%～94.8%),CD49d(16.8%～31.0%)and CD106(8.3%～22.2%).There was a higher CD49d percentage than CD106 in all the passages.The induction efficacy of ADSCs was 17.5%～46.0% for rhodopsin,19.7%～79.3% for CK and 27.3%～50.7% for S-100.Conclusion It is suggested that ADSC has potential to differentiate into photoceptors and RPE cells as evidenced by thepresence of the specific markers of photoceptors(rhodopsin) and RPE markers(CK and S-100).