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.

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.

Objective To investigate the chemical constituents of the 60% alcohol extract of Solanum lyratum Thunb..Method The compounds were isolated by column chromatography over silica gel and Sephadex LH-20 and preparative TLC.Their structures were elucidated on the basis of physicochemical property and spectral data.Resulut Eleven compounds were isolated and identified as:ononin(1), genistin(2), 5-hydroxyl ononin(3), formononetin(4), daidzein(5), daidzin(6), 4-hydroxy-benzaldehyde(7),vanillic acid(8), protocatechuic acid(9),ethyl-α-D-arabinofuranoside(10) and ursolic acid(11).Conclusion Compounds 1,2,3,10 and 11 are isolated from S.lyratum for the first time.

Objective To explore the expression of cyclooxygenase-1 in ovarian cancer and its significance. Methods Expression of COX-1 and CA125 was detected by immunohistochemistry in 37 cases of ovarian cancer and 31 cases of ovarian cyst. The serum level of COX-1 and CA125 was tested in 40 cases of ovarian cancer and 31 cases of ovarian cyst and 60 healthy volunteers by ELISA. Results The positive expression of COX-1 and CA125 was 78% and 57% in ovarian cancer and 3% and 7% in ovarian cyst,respectively. The expression of COX-1 and CA125 was 65% and 93% at the serum level. Conclusion Expression of COX-1 could act as an auxiliary diagnostic criterion in ovarian carcinoma.

Objective To create a model with simple expression of mechanical characteristics of the human chest for the development of a manikin. Methods A simplified Gruben-model was proposed based on the anatomical structure and physical characteristics of the materials, and perfect coefficients were computed. The model feasibility was proved by the coefficient of determination and residual analysis.Results The mathematic form of the model provided had three fewer terms than Gruben′s. The coefficient of determination approximated 1, the residue was small, and the perfect coefficients of "a typical human" were determined.Conclusion The hypothesis of the model makes the coefficients physically meaningful, which provides a new method to study the force-displacement relationship of the thorax. Also the simple form makes it easy to create the model and provide some guidance for the design of a manikin′s chest.

Objective To explore the effect of recovery sleep on the executive function after 36 h of total sleep deprivation by event related potential technology.Methods Thirteen healthy male college students participated in two trials. At the first trial normal sleep as control was investigated. At the second trial participants experienced 36 h of sleep deprivation and then accepted 8 h recovery sleep. In each trial six Go/Nogo tests were employed to test the executive control function and the ERP data were recorded. Results There was no statistical difference in behavior and ERP results at each time point as the subjects had normal sleep. After 36 h of sleep deprivation, the behavior results were statistically significant when compared to the baseline. The amplitude and latency of Nogo-N2, Nogo-P3 on Fz electrode, the amplitude and latency of Nogo-P3 on Cz electrode showed statistical significance when compared to the baseline. After 8 h recovery sleep, the average correct reaction time and the Go correct reaction rate had statistical significance compared to 36 h value. The amplitude of Nogo-N2 and Nogo-P3 had no statistical significance compared to the baseline.However,it was of statistical significance[(-6.80 3.95)vs(-3.37 2.63)μV,(10.63±6.62)vs(5.63±5.45)μV,(9.49±7.37)vs(6.08±6.56)μV] compared to 36 h value. The latency of the recovery value of Nogo-N2 and Nogo-P3 was statistically significant[(254.14±15.55)vs(243.08±13.97)ms(382.14±41.07)vs(349.17±30.36)ms,(369.86±26.48)vs(347.48±29.24)ms]compared to the baseline.Conclusion As the time of sleep deprivation is prolonged, the executive function is impaired and the executive function is not completely recovered after 8 h recovery sleep.