Rationally designed mariner vectors for functional genomic analysis of Actinobacillus pleuropneumoniae and other Pasteurellaceae species by transposon-directed insertion-site sequencing (TraDIS)

Comprehensive identification of conditionally essential genes requires efficient tools for generating high-density transposon libraries that, ideally, can be analysed using next-generation sequencing methods such as Transposon Directed Insertion-site Sequencing (TraDIS). The Himar1 (mariner) transposon is ideal for generating near-saturating mutant libraries, especially in AT-rich chromosomes, as the requirement for integration is a TA dinucleotide, and this transposon has been used for mutagenesis of a wide variety of bacteria. However, plasmids for mariner delivery do not necessarily work well in all bacteria. In particular, there are limited tools for functional genomic analysis of Pasteurellaceae species of major veterinary importance, such as swine and cattle pathogens, Actinobacillus pleuropneumoniae and Pasteurella multocida, respectively. Here, we developed plasmids, pTsodCPC9 and pTlacPC9 (differing only in the promoter driving expression of the transposase gene), that allow delivery of mariner into both these pathogens, but which should also be applicable to a wider range of bacteria. Using the pTlacPC9 vector, we have generated, for the first time, saturating mariner mutant libraries in both A. pleuropneumoniae and P. multocida that showed a near random distribution of insertions around the respective chromosomes as detected by TraDIS. A preliminary screen of 5000 mutants each identified 8 and 14 genes, respectively, that are required for growth under anaerobic conditions. Future high-throughput screening of the generated libraries will facilitate identification of mutants required for growth under different conditions, including in vivo, highlighting key virulence factors and pathways that can be exploited for development of novel therapeutics and vaccines.

Study on establishing hypertension animal models by adverse lifestyles and the antihypertensive effect of traditional Chinese medicine

OBJECTIVE Nowadays,with the living habits and diet structure changes,the morbidity and mortality of hypertension has been rising yearly,which induced by adverse lifestyles,excessive alcohol and poor diet.However,the animal models of hypertension resulted from adverse lifestyles have rarely been reported.It is essential to set up some more systematic models to study the characters of hypertension and evaluate the efficacy of traditional Chinese medicine(TCM).METHODS To build the rat model of human lifehabitual hypertension,SD rats had free access to alcohol with high-sucrose-fat diet(AHSFD),alcohol with high-fat diet(AHFD),high-sucrose-fat diet(HSFD),high-fat diet(HFD)and alcohol etc.Various of indicators were detected systematically to explore the features of hypertension models which included blood pressure(BP),liver and kidney functions,the lipid profiles and indexes quantifying TCM symptoms.Then we choose one of the stable and sustained animal models,studying the effects of RPA.E(which extracted from a TCM)on modulating the level of hypertension and the preliminary mechanism in this abstract.RESULTS ①The BP level,serum TC,TG,LDL-C of AHSFD-induced and AHFD-induced rats increased significantly,while the HDL-C reduced in the 4th week.② The BP level HSFD-induced rats increased significantly in the 6th week,serum TC,TG increased in 10 th week.③ The BP level of alcohol-induced rats increased significantly in the 9th week,serum TC,TG increased in 24 th week.④ The level of serum ALT,AST,UA and Cr of all model rats increased significantly after 12 weeks.Meanwhile the microcirculation increased significantly after8 weeks.⑤ After 4-week administration,RPA.E could significantly reduce BP of AHFD-induced hypertensive rats,and could reduce the serum TC,LDL-C,HDL-C,ALT,AST level after 8-week.CONCLUSION The rats induced by AHSFD,AHFD,HSFD and alcohol can raise BP obviously with underlying hepatic and kidney damage,the lipid profiles disorder and TCM symptoms change,while the AH(S)FD-induced hypertension models are earlier and more stable.RPA.E could mildly reduce BP level,and improve the lipid profiles disorders,hepatic damage,which reflects the characteristics of TCM on antihypertensive effects.The above models have their own characteristics,can be used to study the causes and pathogenesis of hypertension complicating metabolic disorder and the related treatment drug screening.

A Challenge to Social Norms-A Positive Perspective of Richard Cory's Death

This paper tries to analyze E.A.Robinson's famous poem Richard Cory from a new perspective. In the poem, Richard Cory pos sesses abundant wealth as well as good manners, which are highly admired by his town people. However, he chooses to end his life by com mitting suicide. Through analyzing this figure, the paper aims to affirm Richard's challenge of traditional values which emphasize material abundance.

A meta-analysis of the treatment of cancer-related insomnia with acupuncture

To systematically evaluate the clinical efficacy and safety of acupuncture in treating cancer-related insomnia.Methods:Randomized controlled trials(RCTs)and quasi-randomized controlled trials of acupuncture in the treatment of cancer-related insomnia were searched by computer in a Pubmed database,and the retrieved literature was screened according to inclusion and exclusion criteria.After the literature screening was completed,two reviewers read the literature titles independently and extracted effective information.The migration risk assessment was performed by Cochrane.All statistical calculation requirements for this study were fully completed by RevMan5.3 software.Results:Among the 68 selected works of literature,3 RCTs were finally included,with a total of 133 patients.The risk of research bias is generally very low.In 3 RCTs,the sleep score of the experimental group was decreased by 0.48 more than that of the control group in 3 RCTs,with a 95%confidence interval of-0.82 and-0.13,and the degree of decrease was statistically significant(Z=2.70,P<0.05).Conclusion:Higher levels may indicate that acupuncture-related therapies are more effective in improving cancer-related insomnia.Although the included literatures feature low bias and no heterogeneity,the number may have reduced its clinical significance.Therefore,more studies on high-quality randomized controlled trials should be conducted in the future to better clarify the therapeutic effect of acupuncture and moxibustion on cancer-related insomnia.

Contrasting cognitive behavioral therapy for insomnia to analyze sleep theory in Laolao Hengyan

Insomnia is a prominent problem.The current mainstream therapy for insomnia is cognitive behavioral therapy.Traditional Chinese medicine has rich clinical experience in treating insomnia,but we have ignored the contribution of traditional Chinese medicine to behavioral therapy in the past.Completed in the Qing Dynasty in China,Laolao Hengyan is a comprehensive book on the management of the elderly's self-health.The book describes many ways to promote sleep health.This book inherits the traditional Chinese medical treatment concept and reflects traditional Chinese culture.By comparing this book with cognitive behavioral therapy,we find that the Chinese Qing Dynasty literature has documented some cognitive and behavioral aspects of sleep that are consistent with modern medical concepts.Moreover,this method is superior to modern medicine in some aspects,so this is worthy of our in-depth study to complement and enrich modern medical theory.

Chronicle for Chemical Fertilizers in 2009

First,China started to practice market-orientedpricing system for the chemical fertilizersector,effective January 25th,2009.(CCR2009 No.2,No.4) Thereafter,the pricesof domestically made fertilizers and importedfertilizers excluding imported potash fertilizerwill be set by the market itself.The governmentcanceled control over the tradingprices.This reform brought a great impact onthe chemical fertilizer industry and