Objective To investigate the expression of Foxp3~+ lymphocytes in breast carcinoma tissues and their correlation with other pathological factors,and to investigate the mechanism of action of Treg cells.Methods The expression of Foxp3~+ lymphocytes in the breast cancer tissue and non-cancerous tissue was detected by flow cytometry (FCM) in 30 breast carcinoma patients, and its correlation with other pathological factors was statistically analyzed by multiple linear regression analysis.The expression of TGF-β and IL-10 in the lymphocytes infiltrated in breast cancer tissue and non-cancerous tissue was measured by immunohistochemistry, and their correlation with the expression of Foxp3~+ lymphocytes was statistically analyzed by linear correlation dependability analysis. Results There was significant difference in the expression of Foxp3~+ lymphocytes between the malignant and non-cancerous breast tissues(P<0.05),and it was positively correlated with the clinical stage,blood vessel invasion and the matter of axillary lymph node metastasis(P<0.05). The expression of IL-10 in the tumor infiltrating lymphocytes was positively correlated with the expression of Foxp3~+ lymphocytes(P<0.05).Conclusion The expression level of Foxp3~+ lymphocytes is correlated with invasion and metastasis of breast carcinoma, and the IL-10 secreted by Foxp3~+ lymphocytes may be involved in this effect.Foxp3~+ lymphocytes can be used as an assistant marker for prediction and new therpeutic target of breast cancer.

Objective To establish a euglycemic clamp technique in beagle dogs. Methods The euglycemic clamp technique was applied in healthy beagle dogs and the blood glucose, insulin, C-peptide, insulin and glucagon were monitored during the clamp. Results The blood glucose was controlled within the basal level.The coefficient of variation was less than 5% during the clamp. The serum insulin concentration finally reached(40.0±3.8)mIU/L stably and a significant inhibition was shown in endogenous insulin by the determination of C-peptide. But there was no significant increase in serum glucagon compared with basal values.Conclusion Methodology confirmed that the euglycemic clamp technique is successful in beagle dogs and can be applied in the study of pharmacodynamics of insulin preparations.

Objective To develop a sensitive,specific, simple and rapid quantitative real-time PCR (Q-PCR) assay for detection of Staphylococcus aureus with SmartCycler.Methods According to the nuc gene sequences specific to S.aureus, a pair of primers and one TaqMan probe were designed. An internal amplification control (IAC) which is a chimeric double-stranded DNA constructed from a fragment of the Listeria monocytogenes hly gene flanked by the nuc-specific target sequences was added to the reaction system. This IAC was detected using a second TaqMan probe labeled with a different fluorophore. The performance of the nuc-IAC Q-PCR was evaluated using artificially contaminated drinking water and commercial UTH whole milk samples spiked with ATCC 6538.Results The nuc-IAC assay could be used reliably for detection with a sensitivity of 5 copies of linear plasmid DNA per reaction, 10 fg of genomic DNA in 62.5% of the reactions or 50 cfu/ml S.aureus cells with 50% probability. The quantification was linear (r~2≥0.998) over a 6-log dynamic range, with a PCR efficiency over 0.967. The 5×10~2 CFU per 25 ml mimic sample of drinking water or milk could be detected by this assay consistently and quantifiably.Conclusion The nuc-IAC Q-PCR assay for S.aureus is developed. It could not only be applied for the quantitative detection of S.aureus, but also prevent the false negatives and underestimations of contamination loads due to PCR failure.

Objective To study the expression of the fused proteasome activator REGγ(11S regulator complex gamma subunit) using gene recombination technology and to further study the interaction between REGγ and casein kinase-2 interacting protein-1(CKIP-1)in vitro.Methods Firstly, the full length cDNA fragment of REGγ was amplified through PCR using the plasmid pCMV-Myc-REGγ as template and subcloned into the prokaryotic expression vector pGEX-4T-2 before being transformed into E.coli BL21 cells. The protein expression was induced by isopropyl-β-D-thiogalactoside(IPTG) .Secondly, the protein expression was monitored by SDS-PAGE and Western blotting after ultrasonication. Finally, the GST Pull-down assay was performed to investigate the interaction between REGγ and CKIP-1 in vitro.Results The prokaryotic expression construct pGEX-4T-2-REGγ was generated successfully and confirmed by DNA sequencing. Expression analysis showed that the GST-REGγ protein was easily expressed and isolated mainly in the lysate supernatant after sonication and centrifugation. The GST Pull-down assay revealed the strong mutual interaction between REGγ and CKIP-1 in vitro.Conclusion The proteasome activator REGγ could interact with the negative regulator of osteoblastogenesis CKIP-1 in vitro and the current study has shed light on further investigations of their physiological relevance.

S100A4, a member of S100 superfamily of Ca~(2+)-binding proteins, is a polypeptide containing 101 amino acids. S100A4, which is overexpressed in most tumor cells, plays pivotal roles in growth, invasion and migration of tumor by regulating cell cycle progression, differentiation, apoptosis, invasion and metastasis of cells. This article reviews S100A4′s structure, function and potential application as the new target in the therapy of tumors.

Immune and tissue cells usually express pattern-recognition receptors (PRRs) to detect viruses and other microorganisms, thereby inducing signal cascade amplification and host innate immune responses. Since PRRs have strain-specific substrates and mechanisms of recognition, the identification of PRRs and mechanisms of PRRs-mediated responses is highly challenging. Besides, the research on RLRs-mediated immune responses has become more popular in cellular immunology recently. Accumulating evidence shows that post-translation modifications, such as ubiquitination, deubiquitination and ISGylation, play an important role in regulating host innate immune responses. In parallel, these approaches may be used by viruses to evade PRRs-mediated responses or to actively subvert these pathways for their own benefit. It was identified that STING (also called MITA/MPYS/ERIS) plays an important role in RIG-Ⅰ-like receptor(RLR) signaling as a type Ⅰ IFN stimulator, providing a special method for the research on complex host antiviral innate immune responses.

Heterogeneous nuclear ribonulcleoprotein(hnRNP)plays a variety of roles in pre-mRNA splicing, nuclear export of mRNA and turnover. It is reported that hnRNP not only takes part in regulating the development of neuron and glial cells, but is closely related to various central nervous system diseases. This paper reviewed the structure, function and the role of hnRNP in the central nervous system in order to provide new insight into the molecule mechanism of nervous system diseases.

Multilocus sequence typing (MLST) is a molecular genotyping method based on nucleotide sequencing. The procedure of this method characterizes isolates of bacterial species using the DNA sequencing of multiple housekeeping genes(usually seven). For each housekeeping gene, the different sequences present within a bacterial species are assigned as distinct alleles.For each isolate, the alleles at each of the loci define the allelic profile or sequence type (ST). MLST has the advantages of being robust (based on genetic data) and electronically portable to generate data that allow rapid and global comparisons between different laboratories. In this paper, the principle, method, data analysis, application, advantages and flaws of MLST are introduced.

Objective To analyze the frontier and evolution of cognitive neuroscience. Methods CiteSpace Ⅱ was running on Java platform. Results By analyzing the node types,such as the author\institution\country\keyword\cited reference with the tool of Information Visualization, the critical subject for reference and the focuses were quickly defined, and the basic literature and the key literature of cognitive neuroscience were specifically determined and carefully reviewed.Conclusion Keeping track of the evolution trends and significant changes will contribute to development of cognitive neuroscience.

Objective To investigate the chemical constituents of Rannunculus chinensis Bunge..Methods The chemical constituents of R.chinensis were isolated by chromatography on silica gel and Sephadex LH-20. The structures of compounds were identified by phytochemical properties and spectral analysis(MS and NMR).Results Ten compounds were isolated and identified as quercetin(1),kaempferol(2),luteolin(3),quercitrin(4), protocatechuic acid(5),gallic acid(6),ellagic acid(7),kaempferol-3-O-β-rutinoside (8),β-sitosterol (9) and 7-ketologanin (10). Conclusion Compound 10 is isolated from R.chinensis for the first time. Compounds 1,2,3,4,5,6,7 and 8 are obtained from the title plant for the first time.