AIM: Recently,it is widely accepted that atherosclerosis(AS) is an auto-immune related disease and the oxidized-low density lipoprotein(ox-LDL) is the most important AS-related antigen.In order to prevent immune injuries in AS and find new strategies to prevent AS,the immune tolerance of T cells to ox-LDL in vitro was induced in this study.METHODS: Human monocytes were separated from peripheral blood to induce dendritic cells(DCs).DCs were treated with LPS(30 ?g/L),ox-LDL(10 mg/L) and LDL(10 mg/L) for 48 h.Then DCs were mixed with allogenic T lymphocytes to carry out mixed lymphocytes reaction(MLR).CTLA4Ig in different concentrations was added in the MLR of ox-LDL group.MTT method was used to assay the proliferation of T cells and expressed in stimulation index(IS).The CD25 expression and apoptosis of T cells in MLR were tested by flow cytometry.The excretion of IL-2,IFN-? and IL-4 was assayed by ELISpot method.RESULTS: SI in ox-LDL group was higher than that in LDL group significantly(P

Objective To induce the immunoincompetence of T cells to oxidized-low density lipoprotein(ox-LDL)in vitro,in order to prevent immune injuries in atherosclerosis(AS)and to find new strategies to prevent AS.Methods Monocytes were separated from peripheral blood to induce dendritic cells (DC).DCs were treated with LPS(30 ng/m1),ox-LDL(10μg/m1)and LDL(10μg/m1)for 48 h,respectively.Then DCs were mixed with allogeneic T lymphocytes to earry out mixed lymphoeytes reaction (MLR).CTLA4Ig in different concentrations was added into the MLR of ox-LDL group.MTr method was used to assay the proliferation of T cells.The CD25 expression and apoptosis of T cells in MLR were tested by flow cytometry.And the excretion of IL-2,IFN-γ and IL-4 were assayed by ELISPOT method.Results SI of the ox-LDL group was higher than that of the LDL group significantly(DC:T=1:5,1.6717±0.3152vs 1.4250±0.2874.P<0.05;DC:T=1:10,1.5458±0.2748 vs 1.3352±0.2991,P<0.05),and CTLA4Ig inhibited the SI of the ox-LDL group in dose-dependence(CTLA4Ig 1.25μg/ml,0.96±0.30 vs μg/ml 1.29±0.28 vs 1.64±0.33 P<0.05).CTLA4Ig caused the decrease of CD25 expression(CTLA4Ig 1.25 μg/ml,11.26±0.58 vs 14.25±1.02,P<0.05;CTLA4Ig 10μg/rnl 8.42±0.45,P<0.01)and induced apoptosis of T cells in MLR(CTLA4Ig 1.25μg/ml,12.54±3.69 vs 6.09 4-2.24,P<0.05;CTLA4Ig 10μg/ml,26.87±5.06 VS 6.09±2.24,P<0.01).CTLA4Ig caused the decrease of ELISPOT counts of IL-2(CTLA4Ig 1.25μg/ml,386±42 VS 534±54,P<0.05;CTLA4Ig 10μg/rnl,230±27 VS 534±54,P<0.01)and IFN-γ(CTLA4Ig 1.25μg/ml,445±48 VS 672±46,P<0.05;CTLA4Ig 10μg/ml,193±39 VS 672±46,P<0.01),while that of IL-4 increased(CTLA4Ig 1.25μg/ml,401±32 VS 332±41,P<0.05;CTLA4Ig 10μg/ml,453±57 VS 332±41,P<0.05).Conclusion CTLA4Ig can induce T cens immunoin competence to ox-LDL in vitro by inhibiting T cells activation,inducing T cells apoptosis and TH 1/TH2 immune deviation.

AIM : To set up a method of determinting the contents of four iridoid glycosides in different preparations of Lamiophlomis rotata ( Benth.) kudo.METHODS : The quantitative analysis was carded out on a column of Waters Symmetry by HPLC using a mobile phase of CH_3OH-H_2O under a flow rate of 1.0 mL/min.235 nm was selected as the detective wavelength.RESULTS: The linear ranges were 0.236-1.18 μg for sesamoside,0.440-2.20 μg for shanzhiside methyl ester,0.094-0.470 μg for 7,8-dehydropenste moside、0.750-3.75 μg for 8-O-ace-tylshanzhiside methyl ester.The recoveries of them were all between 96% and 104% ( RSD ＜ 5% ).CONCLUSION: The method is accurate,stable,reliable,and can be used as a quantitative standard for determining the contents of iridoid glycosides in different Duyiwei preparations.

AIM:To set up a method of determinting the contents of four iridoid glycosides in different preparations of Lamiophlomis rotata(Benth.)kudo.METHODS:The quantitative analysis was carried out on a column of Waters Symmetry by HPLC using a mobile phase of CH_3OH-H_2O under a flow rate of 1.0 mL/min.235 nm was selected as the detective wavelength.RESULTS:The linear ranges were 0.236-1.18 ?g for sesamoside,0.440-2.20 ?g for shanzhiside methyl ester、0.094-0.470 ?g for 7,8-dehydropenste moside、0.750-3.75 ?g for 8-O-acetylshanzhiside methyl ester.The recoveries of them were all between 96% and 104%(RSD

OBJECTIVE:To develop a method for simultaneous determination of 6 alkaloids in Coptis teeta.METHODS:The determination was performed on XtimateTM C18 with mobile phase consisted of 30 mmol/L ammonium bicarbonate [containing 0.1％ triethylamine and 0.7％ ammonia-acetonitrile (gradient elution)] at the flow rate of 1.0 mL/min.The detection wavelength was set at 270 nm,the column temperature was 30 ℃,and the sample size was 10 μL.RESULTS:The linear ranges of jateorrhizine,columbamine,epiberberine,coptisine,palmatine hydrochloride and berberine hydrochloride were 0.85-16.96 mg/L (r=0.999 9),1.25-24.96 mg/L(r=0.999 8),2.05-40.96 mg/L(r=0.999 9),3.65-72.96 mg/L(r=0.999 9),2.88-57.60 mg/L(r=0.999 9) and 13.25-264.96 mg/L(r=0.999 9),respectively.RSDs of precision,stability and reproducibility tests were all lower than 3.0％.Recoveries were 97.14％-102.14％ (RSD=1.93％,n=6),97.00％-102.00％ (RSD=2.06％,n=6),98.18％-101.82 ％ (RSD=1.79％,n=6),96.15％-101.28％ (RSD=2.06％,n=6),96.88％-101.88％ (RSD=1.87％,n=6),99.31％-103.76％ (RSD=1.89％,n=6),respectively.CONCLUSIONS:The method is simple,accurate,stability and reproducible,and can be used for simultaneous determination of 6 alkaloids in Coptis teeta.

Objective To investigate the genotypes of host killing genes and their single nucleotide polymorphisms (SNPs). Methods Three hundred and twenty strains of Escherichia coli that collected from the First Affiliated Hospital of Wenzhou Medical College were analyzed. The first sample ( E1 ) contains 160 strains isolated during the years from 2002 to 2003. The second sample (E2) contains 160 strains covering the years from 2008 to 2009. The plasmids of Escherichia coli were extracted by alkaline lysis method. Solexa/Illumina sequencing technology was used to sequence plasmids metagenome. Solexa Genome Analysis System and Soap programs were used to analyze gene distribution, SNPs and lineage-specific mutations. Results 11 077 768 reads were generated and 0. 045% of them can map to the reference sequences from El sample. Whereas 9 377 792 reads were generated and 0. 053% of which mapped to the reference from E2 sample. There are nine host killing genes identified in the two samples, of which hok gene is the most prevalent. A total of 29 SNP sites dispersed in five genes of the two samples. Approximately 33% of them were non-synonymous mutations. One position of A and G is the most prevalent polymorphism. Conclusion The known nine genotypes of host killing genes were all identified in plasmids of Escherichia coli in Wenzhou. hok gene showed the highest frequency. There were SNPs in five genotypes.

Objective To establish the HPLC fingerprints of Tangbikang Granules; To scientifically evaluate and effectively control the quality of Tangbikang Granules; To ensure its production stability. Methods HPLC was performed on the column of Germany Merck RP-18 endcapped (250 mm × 4.6 mm, 5 μm) with mobile phase of acetonitrile-0.1% formic acid water; column temperature was 40 ℃; flow rate was 1.0 mL/min; detection wavelength was 240 nm; volume injection was 20 μL. Fingerprint Similarity Evaluation Software (edition 2004A) of Chinese Pharmacopoeia Commission was used to evaluate the similarity of the 10 batches of Tangbikang Granules, and to analyze the correlations of 9 ingredients in Tangbikang Granules. Results Wogonoside was used as the reference peak, and the common mode for the HPLC fingerprints was set up. The similarities of the 10 batches of Tangbikang Granules were above 0.930, and altogether 25 common peaks in the chromatograms were found, of which 18 peaks were assigned to Chinese materia medica in Tangbikang Granules. Conclusion The method has good separability and is accurate and simple, which can provide references for the quality control of Tangbikang Granules.

Objective:To investigate the influence of abnormal blood pressure circadian rhythms and the morning surge on aortic stiffness in elderly patients with isolated systolic hypertension(ISH).Methods:A total of 300 untreated elderly(≥60 years)patients with ISH were enrolled.24-hour ambulatory blood pressure was measured by using a non-invasive portable ambulatory sphygmomanometer with an inflatable cuff.Brachial-ankle pulse wave velocity(baPWV)and the ankle-brachial pressure index(ABI)were measured by using an automated device.Patients were divided into the dipper(n=95), no-dipper(n=177)and extreme dipper(n=28)groups according to the rate of nocturnal blood pressure reduction.They were also divided into the morning surge(n=88)and no morning surge(n=212)groups according to the morning blood pressure surge(MBPS). Differences between the groups were compared.Correlations between the parameters were calculated by partial correlation analyses.The effects on baPWV and ABI were calculated by multiple linear regression analyses.Results:baPWV was higher in the extreme dipper group than in the dipper and no dipper groups[(1 402±234)cm/s vs.(1 467±114)cm/s vs.(1 538±140)cm/s, P<0.01], while ABI values were lower in the extreme dipper group than in the dipper group(0.98±0.10 vs.1.05±0.12, P<0.01)and the no dipper group(0.98±0.10 vs.1.03±0.12, P<0.05). Moreover, baPWV[(1 508±170)cm/s vs.(1 430±163)cm/s, P<0.01]was higher while ABI values(0.98±0.13 vs.1.06±0.11, P<0.01)were lower in the morning surge group than in the no morning surge group.baPWV was positively correlated with daytime Systolic blood pressure(dSBP)( r=0.169, P<0.01), 24 hSBPCV( r=0.143, P<0.05), and MBPS( r=0.157, P<0.01), while ABI was negatively correlated with dSBP( r=-0.146, P=0.011)and MBPS( r=-0.321, P<0.01). Age( P<0.01), dSBP( P<0.05)and 24 h systolic blood pressure variation coefficient( P<0.05)were independent factors for baPWV, while dSBP and MBPS were independent factors for ABI(all P<0.01). Conclusions:Abnormal blood pressure circadian rhythms and the morning surge are associated with increased aortic stiffness in elderly patients with ISH.

Amyloid deposits are one of the hallmark pathological lesions of Alzheimer's disease (AD). They can be visualized by thioflavin-S, silver impregnation, Congo red staining, and immunohistochemical reactions. However, that amyloid deposits generate blue autofluorescence (auto-F) has been ignored. Here, we report that visible light-induced auto-F of senile plaques (SPs) was detected and validated with conventional methods. Brain slices from APP/PS1 (amyloid precursor protein/presenilin 1) transgenic mice were mounted on slides, rinsed, coverslipped and observed for details of the imaging and spectral characteristics of the auto-F of SPs. Then the slices were treated with the above classic methods for comparative validation. We found that the SP auto-F was greatest under blue-violet excitation with a specific emission spectrum, and was much easier, more sensitive, and reliable than the classic methods. Because it does not damage slices, observation of auto-F can be combined with all post-staining techniques in slices and for brain-wide imaging in AD.