Objective To establish a real-time quantitative PCR ( qPCR) method for detection of Streptobacillus moniliformis, which can be used to rapidly detect this pathogen in laboratory animals .Method According to the S. moniliformis sequences published in NCBI , we designed specific primers and MGB probe .The specificity, sensitivity and stability of this method were evaluated using 24 standard reference strains .Total of 823 respiratory specimens of animals including mice, rats, guinea pigs, hamsters, rabbits, Mongolian gerbils and tree shrews , were detected by this established Taqman MGB qPCR method .Results We had successfully established the S.moniliformis Taqman MGB qPCR method . S.moniliformis was not detected in the samples of mice , rats, guinea pigs, hamsters and rabbits.The positive rate of S. moniliformis was 1.5% ( 1/65 ) and 61.7% ( 37/60 ) in conventional Mongolian Gerbils and tree shrews , respectively . Conclusions Our developed qPCR method can be used to effectively detect S.moniliformis in laboratory animals .Moreover , its accuracy and sensitivity are better than the national standard method .This study laid the foundations for optimizing the quality inspection system of laboratory animals .

Objective We established a rapid detection method of Pasteurella spp.and provided a reference for microbiological quality control of laboratory animal .Methods According to the β subunit of bacterial RNA polymerase ( rpoB) protein multiple alignments of 13 different Pasteurella spp.published in NCBI .The degenerate primers were designed by CODEHOP designer online .CODEHOP PCR method was applied to detecting Pasteurella spp.after the specificity and sensitivity of the method had been evaluated by 21 reference strains .Results Standard strain amplified fragment were about 200 bp by degenerate primers PastF6/PastR5.The primers are able to distinguish between Pasteurella spp.and the other pathognic organisms of laboratory animal respiratory tracts .Sensitivity of this method were 0.2 pg/μL~2 pg/μL to different Pasteurella.The Pasteurella positive rate was 19.1% in 609 animal ' s respiratory samples .The accuracy of positive results was 100%through verifying by sequenced and blast .Conclusions The established method has good specificity and sensitivity .It can be used to detect Pasteurella spp.in animal samples .

The effects of Sonic hedgehog (Shh) signaling pathway activation on S-type neuroblastoma (NB) cell lines and its role in NB tumorigenesis were investigated. Immunohistochemistry was used to detect the expression of Shh pathway components-Patched1 (PTCH1) and Gli1 in 40 human primary NB samples. Western blotting and RT-PCR were used to examine the protein expression and mRNA levels of PTCH1 and Gli1 in three kinds of S-type NB cell lines (SK-N-AS, SK-N-SH and SHEP1), respectively. Exogenous Shh was administrated to activate Shh signaling pathway while cyclopamine was used as a selective antagonist of Shh pathway. S-type NB cell lines were treated with different concentrations of Shh or/and cyclopamine for different durations. Cell viability was measured by using MTT method. Apoptosis rate and cell cycle were assayed by flow cytometry. The xenograft experiments were used to evaluate the role of Shh pathway in tumor growth in immunodeficient mice. High-level expression of PTCH1 and Gli1 was detected in both NB samples and S-type NB cell lines. Cyclopamine decreased the survival rate of the three cell lines while Shh increased it, and the inhibition effects of cyclopamine could be partially reversed by shh pre-treatment. Cyclopamine induced the cell apoptosis and the cell cycle arrest in G(0)/G(1) phase, while Shh induced the reverse effects and could partially prevent effects of cyclopamine. Cyclopamine could also inhibit the growth of NB in vivo. Our studies revealed that activation of the Shh pathway is important for survival and proliferation of S-type NB cells in vivo and in vitro through affecting cell apoptosis and cell cycle, suggesting a new therapeutic approach to NB.

The peripheral blood lymphocytes (PBLs) from 111 cancer patients were isolated and cultured respectively for 23 - 27 days in the medium mainly conditioned by IL-2 and PHA. With ~(125) I-UdR release method, sampling in random way, we examed the cytotoxicities of PBLs and lymphokine-activated-killer (LAK) cells in different culture periods in vitro. The statistic analysis on sufficient data gave the following results: 1. The cytotoxicity against K562 cells increased from 34.78 ?25% of the PBLs to 68.04 ?17.3% of the cells cultured for 8-13 days, afterward, kept about 70% to 23 - 27 days. The constitutional proportion patterns showed that the freshly isolated samples dispersed at a wide range of cytotoxicities (10 - 90%), and that most of the cultured samples ( ~ 85%) concentrated on the range of higher cytotoxicities (50 ~ 95% ) after 8-13 days. 2. The cytotoxicity against Raji cells rose from 8.9 ?8% of the fresh PBLs to 42.1 ?22% of the LAK cells at 8 - 13 days, and maintained about 35% in the following periods. The constitutional proportion patterns of the cytotoxicity against Raji illustrated that all the fresh PBL samples were below 25% of cytotoxicity, and that during the culture, one part of the samples ( ~ 30%) acquired the higher cytotoxicities (50 -90% ), but the other part of the samples ( - 40%) remained at lower cytotoxicities (below 35% ) . The mechanisms behind these phenomena are worth further investigating.

Objective:To summarize the evidence for the link between rheumatoid arthritis and risk of vertebral fractures or vertebral deformities with a meta-analysis, so as to provide objective proof for early preventing and the development of vertebral deformity and fractures.Methods:Wanfang,CNKI,VIP,PUBMED,Springlink and Elsevier were retrieved for all publications relating to rheumatoid arthritis and vertebral fractures in women.According to inclusion and exclusion criteria,two investigators collected their data individually,then statistical analysis was performed using Stata 12.0 software.Results: Eight case-control studies were enrolled, including 86 741 participants,2 258 of them with RA.The results of Meta-analysis showed that a higher incidence of vertebral fractures in RA,and Odds ratio was 3.70 with a 95%confidence interval(2.47-5.55,P<0.000 1).The publication bias analysis did not reveal any evidence of obvious asymmetry, and the sensitivity analysis showed that omission of any individual study made no significant difference for all comparison models,suggesting that our results were statistically robust.Conclusion:RA may be one of the risk factors for the vertebral fractures.

Objective To improve hand hygiene compliance among health care workers(HCWs)through continu-ous quality improvement,and effectively reduce the incidence of healthcare-associated infection(HAI).Methods Continuous quality improvement was performed by adopting plan-do-check-action(PDCA)cycle,all HCWs were trained,hand hygiene was stressed,periodical and random checking was conducted.Results After the implementa-tion of PDCA cycle,the acknowledge rate of hand hygiene enhanced from 48.00% to 63.99%;hand hygiene com-pliance rate enhanced from 65.11% to 82.40%,the difference were both significant(χ2=12.75,259.65,respective-ly,both P<0.05).The daily consumption of instant hand antiseptic per 1 000 bed day increased obviously,which was 2.95-fold of pre-implementation.Conclusion Continuous quality improvement through PDCA cycle can effec-tively improve hand hygiene compliance rate of HCWs.

Objective To investigate the association between levels of serum alanine aminotransferase (ALT) and the risks of metabolic syndrome in middle-aged and elderly Chinese. Methods After excluding subjects with known liver disease, excess alcohol consumption and serum ALT≥40 IU/L,1 664 subjects aged 40 years or older from Baoshan Community, Shanghai were recruited to undergo questionnaire interview, anthropometric measurements, and fasting blood sampling. Biochemical features were evaluated and the metabolic syndrome was defined according to the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) criteria with modification on waist circumference cutoff which is more appropriate for an Asian population. Results Increased ALT levels were associated with more unfavorable metabolic risk profiles. The prevalence rates of the metabolic syndrome in participants with increasing ALT quartiles were 29. 2% , 38. 0% , 44.9% , and 62. 9% , respectively (P for trend <0. 01 ). ALT concentrations were significantly elevated with increasing number of the metabolic syndrome components (P for trend<0.01). Serum ALT levels were significantly associated with the risks of metabolic syndrome and most of its components in a dose-response manner. As compared with participants in the first ALT quartile, the risks of metabolic syndrome were increased by 146% , central obesity by 204% , hypertension by 35% , high triglycerides by 133% , and hyperglycemia by 72% in participants of the fourth ALT quartile. Conclusions A high-normal serum ALT level was significantly associated with an increased risk of the metabolic syndrome in middle-aged and elderly Chinese.

The effects of Sonic hedgehog (Shh) signaling pathway activation on S-type neuroblastoma (NB) cell lines and its role in NB tumorigenesis were investigated. Immunohistochemistry was used to detect the expression of Shh pathway components-- Patchedl (PTCH1) and Glil in 40 human primary NB samples. Western blotting and RT-PCR were used to examine the protein expression and mRNA levels of PTCH1 and Glil in three kinds of S-type NB cell lines (SK-N-AS, SK-N-SH and SHEPI), re-spectively. Exogenous Shh was administrated to activate Shh signaling pathway while cyclopamine was used as a selective antagonist of Shh pathway. S-type NB cell lines were treated with different concen-trations of Shh or/and cyclopamine for different durations. Cell viability was measured by using MTT method. Apoptosis rate and cell cycle were assayed by flow cytometry. The xenograft experiments were used to evaluate the role of Shh pathway in tumor growth in immunodeficient mice. High-level expres-sion of PTCH1 and Gill was detected in both NB samples and S-type NB cell lines. Cyclopamine de-creased the survival rate of the three cell lines while Shh increased it, and the inhibition effects of cyclopaminc could be partially reversed by shh pre-treatment. Cyclopamine induced the cell apoptosis and the cell cycle arrest in G0/G1 phase, while Shh induced the reverse effects and could partially pre-vent effects of cyclopamine. Cyclopamine could also inhibit the growth of NB in vivo. Our studies re-vealed that activation of the Shh pathway is important for survival and proliferation of S-type NB cells in vivo and in vitro through affecting cell apoptosis and cell cycle, suggesting a new therapeutic ap-proach to NB.

Objective To analyze the differential gene expression between thymic carcinoids and normal thymic tissues and to study thepathogenesisofectopicACTH syndrome caused by thymic carcinoids. Methods Using gene chip technique, the gene differential expression of 2 tissues were observed following RNA labeled with different fluorescences (Cy3 and Cy5) hybridized to gene chip. Results Among 4224 genes on gene chip, 394 were up regulated more than 2 folds in thymic carcinoid tissues, 23 of which were associated with cell mitosis; 51 genes were upregulated more than 5 folds, 1 of which (PAK3) was associated with cell mitosis. Conclusion A group of differentially expressed genes were observed between the thymic carcinoids and normal thymic tissues.These overexpressed and cell mitosis-associated genes probably play a role in the pathogenesis of thymic carcinoid tumors.

Objective This study aimed to investigate the associations between adipose tissue distribution and risks of macro-vascular complications in type 2 diabetic patients. Methods The study included T2DM patients either previously diagnosed by questionnaire or newly diagnosed with fasting plasma glucose(FPG)≥7. 0 mmol/L and/or 2 hour postprandial glucose ( 2h PG )≥11. 1 mmol/L from residents older than 40 years from Jiading Community, Shanghai, China(n=10 375). Each participant had gone through anthropometric measurements, blood tests, vascular function tests including carotid intima-media thickness ( CIMT ) , ankle-brachial index ( ABI ) , branchial-ankle pulse wave velocity( baPWV) as well as carotid plaques. Results WHR quartiles was significantly associated with high risks for ten-year cardiovascular disease risks [ ASCVD high risk: OR:1. 17 ( 1. 05-1. 31 );Framingham high risk:1. 13(1. 00-1. 29)]. However, WHR has no significant relations with risks of increased carotid intima-media thickness( CIMT) , abnormal ankle-brachial index( ABI) , increased brachial ankle pulse wave velocity( baPWV) or carotid plaques. Conclusions In type 2 diabetic patients, abdominal adipose tissue has an independent role in high risk for ten-year cardiovascular diseases. However it has no relations with other vascular functions including increased CIMT, abnormal ABI, increased baPWV, or carotid plaque.