The complete length of P gene from rabies virus was amplified by RT-PCR using a pair of specific primers designed according to the relevant sequences from GenBank. The PCR product was cloned into cloning expression vestor pGM-T to obtain the cloning expressed plasmid pGM-T-P. After double-digestion by NotI and EcoRI, the product was transferred into prokaryotic expression vetor pET-32a(+)to obtain the prokaryotically expressed plasmid pET-32a-P. The target gene was then expressed in the E.coli BL21(DE3) cell with IPTG induction. The highest expression of target protein was analysed by SDS-PAGE, and the good immunoreactivity to rabies virus antibodies was proved by Western-blot analysis. By using purified protein, the indirect ELISA assay for the detection of rabies virus antibodies in canine serum was applied after management of the optional working condition.

ObjectiveTo investigate the effects of nicotine exposure and ethanol-preferring behavior on mRNA expression of some nAChR subunits in the rat ventral tegmental area(VTA) and to explore possible mechanisms of dependence on tobacco and alcohol.Methods39 male Wistar rats,35-day-old,were randomly divided into an experimental group (group N,n=20) and a control group (group C,n=19).Rats in group N were treated with nicotine ( 1.0 mg · kg -1 · d -1 ) by subcutaneous injection while in group C with saline both for 10 days,after which 6 rats (respectively group NE,n =6,group CE,n =6 )were drawn randomly from each group and killed by cutting off the head.mRNA was extracted from the VTA tissue,and the expression of nAChR subunits,including α4,α5,α7 and β2,were examined by Real Time-PCR.Other rats both in groups N and C ( respectively group NA,n=14,group CA,n=13) were induced for 69 days to establish two-bottle free choice alcohol-preferring behavior model by Samson sucrose fading program from 60-day-old on.The same indexes mentioned above were detected by the same methods in the VTA tissue.Results① The factor analysis showed that both the two factors,nicotine and alcohol-preferring behavior,showed regulating effects on the expression of nAChR subunits α4 and α5 ( respectively F was 6.13,5.407,5.186,7.132,P ＜ 0.05 ),and the factor,alcohol-preferring behavior,on subunit β2 (F =5.896,P＜0.05) ; the two factors exhibited strong interaction on the expression of subunit α7 (F=13.894,P＜0.001 ),and some interaction on subunits α5 and β2 (respectively F was 6.149,4.222,P＜0.05 ).② The mRNA expression of nAChR subunits α4,α5,α7,and β2 were significantly up-regulated by different degrees in group NA compared to group CA ( respectively Fwas 7.941,13.517,17.438,9.272,respectively P ＜ 0.05,P ＜ 0.05,P ＜ 0.01,P ＜ 0.01 ),the expression level of subunits α4,α5,α7 and β2 were significantly higher in different degrees in group NA than in group NE( respectively F was 5.293,8.500,6.149,4.837,P ＜0.05) ; while subunit α7 was significantly down-regulated in group CA compared to group CE (F =12.750,P ＜0.01 ).ConclusionNicotine and ethanol co-affect on the nAChR subtype comprised of subunits α4,α5,α7 and β2.

OBJECTIVE:To investigate the utilization and trend of endocrine therapeutic drugs after breast cancer surgery,and to provide reference for rational drug use in clinic. METHODS:In retrospective study,the data of endocrine therapeutic drugs use in outpatients after breast cancer in a Third Grade Class A Hospital during 2014-2016 was calculated,sorted and analyzed statistically. RESULTS:The age of outpatients receiving endocrine therapy after breast cancer ranged from ＞40 to 60. Among endocrine therapeutic drugs used within 3 years,3 kinds of them were aromatase inhibitor(AI),3 kinds were anti-estrogen and 2 kinds were gonadotropin releasing hormone(GnRH)analogues. The amount of Toremifene citrate tablets(imported)in prescription was much more than the others. The proportion of anti-estrogen drugs alone in prescription was the highest(81.11%). The AI and GnRH analogues were mostly used in drug combination prescription,and commonly combined with anti-osteoporosis drugs. The comsuption sum of endocrine therapeutic drugs was increased year by year,increasing by 52.48% in 2015,40.60% in 2016. The consumption sum of AI ranked the highest proportion among three categories of endocrine drugs(44.71%,48.62%,48.62%,each year respectively). The consumption sum of imported drugs was significantly higher than that of domestic drugs. The consumption sum of Toremifene citrate tablets(imported)increased continuously in 3 years,and took the first place in 2016. The consumption sum of Leuprorelin acetate microspheres for injection(imported)had the largest growth rate when applied to the clinic. Top types in the list of DDC were all imported types;Fulvestrant injection(imported)ranked top 1,and Tamoxifen citrate tablet(domestic) was the bottom of the list. DDDs of endocrine therapeutic drugs also increased year by year,among which Toremifene citrate tablets,Tamoxifen citrate tablets,Anastrozole tablets and Letrozol tablets ranked top 4 places during the 3 years,while Tamoxifen citrate tablets DDDs decreased year by year. B/A values of these drugs were between 0.25 and 9.00. B/A values of domestic varieties were basically more than or equal to 1,of which Tamoxifen citrate tablets was the largest;B/A values of the imported varieties were basically less than or equal to 1. CONCLUSIONS:The consumption sum of endocrine therapeutic drugs after breast cancer surgery increased year by year in 3 years,and DDDs is also increasing year by year. The imported drugs took up the dominant place. Toromiphene citrate tablets had replaced Tamoxifen citrate tablets as the first choice of endocrine therapeutic drugs in the hospital. The frequency of AI and GnRH analogues use had also increased. And the combination of 2 kinds of endocrine therapeutic drugs and endocrine therapeutic drugs combined with anti-osteoporosis drug had become a trend of clinical treatment. The choice of therapeutic drugs should take into account of effectiveness,safety,economics,cost and therapeutic efficacy,so as to guarantee the greatest benefit of the patient.

The self-renewal and multipotent potentials in neural stem cells (NSCs) maintain the normal physiological functions of central nervous system (CNS). The abnormal differentiation of NSCs would lead to CNS disorders. However, the mechanisms of how NSCs differentiate into astrocytes, oligodendrocytes (OLs) and neurons are still unclear, which is mainly due to the complexity of differentiation processes and the limitation of the cell separation method. In this study, we modeled the dynamics of neural cell interactions in a systemic approach by mining the high-throughput genomic and proteomic data, and identified 8615 genes that are involved in various biological processes and functions with significant changes during the differentiation processes. A total of 1559 genes are specifically expressed in neural cells, in which 242 genes are NSC specific, 215 are astrocyte specific, 551 are OL specific, and 563 are neuron specific. In addition, we proposed 57 transcriptional regulators specifically expressed in NSCs may play essential roles in the development courses. These findings provide more comprehensive analysis for better understanding the endogenous mechanisms of NSC fate determination.
Animals ; Astrocytes ; cytology ; metabolism ; Cell Differentiation ; genetics ; Gene Expression Profiling ; Gene Regulatory Networks ; Mice ; Neural Stem Cells ; cytology ; metabolism ; Oligodendroglia ; cytology ; metabolism ; Protein Interaction Mapping