Objective To develop a simple microfluidic chip technology for analyzing the electrotaxis of cancer cells . Methods The basic structure of the proposed microfluidic electrotaxis chip included a straight microchannel and liquid storage pools located on both sides of the microchannel .Two platinum electrodes were inserted into the liquid pools to create a controllable direct current ( DC ) field in the microchannel .The distribution and strength of the DC field in the microchannel was analyzed by the finite element analysis software COMSOL multiphysics and experiment tests .Finally, the electrotactic behavior of the rhabdomyosarcoma RD cells in the DC field of different strength was characterized using the accumulated distance, average velocity, x forward migration index ( xFMI) and y forward migration index ( yFMI) as quantitative parameters.Results The results of element analysis and experiments showed that the structure of the designed microfluidic electrotaxis chip was able to guarantee a uniform and strength-controllable DC field in the microchannel .The experiment of cell electrotaxis showed that the RD cells migrated toward the anode of the DC field .Meanwhile , the values of xFMI and accumulated distance for RD cells increased with the enlargement of the DC field , with the strength ranging from 188 to 1320 V/m.Conclusion The microfluidic chip technology developed in this paper for assaying the electrotaxis of cancer cells is simple and easily implementable , and it can be used for studies of the electrotactic behavior and underlying mechanisms of various cancer cells and normal cells in the future .

AIM:To clarify the mechanism of treating autoimmune cardiomyopathy at different stages with anti-L3T4 monoclonal antibody.METHODS:Mice immunized with human mitochondria ADP/ATP peptides were used as the cardiomyopathy(DCM) group,and the sham-immunized mice were regarded as the controls.Mice receiving early treatment were immunized with the same peptides,followed by the injection of 400 ?g of anti-L3T4 on day 0,1 and 2 post-immunization.Mice in the late treatment group were immunized as of the early treatment group but anti-L3T4 was administered 3 months post-immunization.The cytokine expression was measured with three-color flow cytometry to quantitate the splenic Th1/Th2 cell subsets in the different groups of mice.In addition,serum and myocardial cytokines were measured by enzyme-linked immunosorbent assay and real-time PCR.RESULTS:Th1 and Th2 subsets in the early treatment group were similar to those in control group,but were drastically lower than those in DCM group.Mice in the late treatment group showed an increased level of Th1-related cytokines,while the Th2 level was between the DCM and early treatment group.IFN-? and IL-6 levels in early treatment group were similar to those in control group.In the early treatment group,IL-4 level was higher than that in control and lower than that in DCM group,whereas IL-2 and TNF-? contents were lower than those in control and DCM group.In the late treatment group,IFN-? and IL-2 levels were higher than those in DCM group and lower than those in the early treatment group,while IL-6 and IL-4 levels were lower than those in DCM group.CONCLUSION:These results suggest that the cytokine production in cardiomyopathic mice may be repressed by treatment with anti-L3T4 at different stages.Early treatment with anti-L3T4 has better inhibitory function than treatment in late stage of autoimmune cardiomyopathy.

AIM:We examined the efficacy ofanti-L3T4 McAb in the T cell signaling pathway in treating ex-Derimental antoimmune cardiomyopathy in BALB/c mice,as a model of the autoimmune mechanism involved in human di-latedardiomyopathy(DCM).METHODS:ADP/ATP carrier peptides were used to induce autoimmune cardiomyopathy in BALB/c mice.Afier 3 months.anti-L3T4 McAb WaS administered to deplete CD4+ T ceHs in the mice.Real-time PCR were used to detect the expression of intracellular signaling molecules(p561ck,p59fyn and Zap-70)and cytokine production(IFN-r,IL-2 and IL-4)in T cells.The expression of CIM5 was determined by immunohistochemistry a-nalysis.RESULTS:Reduced expression of p561ck,p59fyn and Zap-70 and the reduced cytokine production of IFN-r, IL-2 and IL-4 in T cells of anti-IJ3T4-treated DCM mice were found.Also,the expression of CD45 in spleen T cells was significantiv decreased in the anti-L3T4-treated group.In contrast,immunization with irrelevant Ab did not protect the mice.the expression of T cell signaling molecules,CD45,and cytokine were not inhibited.CONCLUSION:Thesestudies provide direct evidenee that anti-L3T4 McAb can be all effective immunomodulator to T cell signal molecules and 8ubsequent cytokine production events in ADP/ATP carrier-induced DCM in BALB/c mice.

Objective To evaluate the microstructural integrity of white matter (WM) in mild cognitive impairment ( MCI ) and Alzheimer disease (AD) using DTI technique, and to explore the relationship between WM abnormalities and cognitive dysfunction. Methods Nine cases of amnestic MCI, 15 cases of mild probable AD and 11 cases of normal controls (NC) with normal-appearing WM (NAWM) were studied using 3. 0 T MR system. Fractional anisotropy (FA) and mean diffusivity (MD) were measured in different WM areas. One-way analysis of variance was used to test the difference among the three groups for DTI indices. Spearman Correlation analysis was applied to reveal the correlation between the DTI indices and the MMSE and CASI scores. Results The FA value in parietal, centrum semiovale, posterior cingulate gyrus, parahippocampus, temporal and frontal WM in MCI was 0. 31 ± 0.03,0. 39 ± 0. 03,0. 62 ± 0. 05,0. 59 ± 0. 05,0. 47 ± 0. 08,0. 32 ± 0. 04, respectely, and MD value was ( 899 ± 30 ) × 10-6,(782±53) × 10-6, (732±45) × 10-6, (806±38) × 10-6, (772 ± 55) × 10-6, (792 ± 35) × 10-6 mm2/s. The FA value of these regions in AD was 0. 28 ± 0. 04, 0. 37 ± 0. 03,0. 55 ± 0. 06,0. 52 ± 0.05,0.40±0. 05,0. 27 ± 0. 04,and MD value was (912±37) × 10-6,(800 ± 67) × 10-6, (762 ± 46) × 10-6, (874±57)×10-6,(822±55)×10-6, (822±39)×10-6 mm2/s. The FA value in NC was 0.36±0.03,0.43±0.05,0.64±0.05, 0.60±0.05, 0.52±0.05,0.33±0.03, and MD value was (866±37)×10-6,(754±54)×10-6,(718±32)×10-6,(810±39)×10-6,(755±48) × 10-6, (785±23)×10-6 mm2/s. Compared with NC, the FA value in parietal WM was significantly decreased in MCI(P<0. 01 ), The significantly reduced FA values in parietal, centrum semiovale, posterior cingulate gyrus, parahippocampus, temporal and frontal WM , as well as significantly elevated MD values were found in AD(P <0. 05). There was significant correlation between these DTI indices and MMSE and CASI scores (P<0.05). Conclusions MR DTI can detect WM abnormalities in AD and MCI. The parietal WM abnormalities and the disconnection of WM circuitry may play an important role in the development of dementia.

Objective To investigate the association of multi-modality neuroimaging features and cognitive function in mild cognitive impairment (MCI) and Alzheimer's disease (AD).Methods Nine individuals with amnestic MCI (aMCI), fifteen patients with mild probable AD, and eleven age-controlled cognitively normal controls (NC) were recruited.All participants were administered with mini-mental status examination (MMSE) and Cognitive assessment screening instrument (CASI) to assess general cognitive function.Optimized voxel-based morphometry ( VBM ) was used for the analysis with 3-D high resolution anatomical images.Values of fractional anisotropy (FA) and mean apparent diffusivity coefficient (ADC) were measured from different brain regions on diffusion-tensor images ( DTI) .The relationship between structural atrophy and DTI-based measurements in the selected brain regions was examined.Results The scores of MMSE and CASI were correlated with the volumetric changes in such areas as temporal, frontal and parietal lobes, and cingulate gyrus and hippocampal gyrus (P <0.001).The scores of MMSE and CASI were positively correlated with FA values, and negatively with ADC values in the white-matter-affected regions including temporal, frontal, parietal lobes, cingulate gyrus, and parahippocampal gyrus (P < 0.05).Conclusions Cognitive decline was associated with atrophy and white matter microstructural alterations in temporal, frontal, parietal lobes, cingulate gyrus, and parahippocampal gyrus in MCI and AD. Multi-modality imaging technique may be important in elucidating the brain mechanism of cognitive impairment.

The cytokine repertoire of ADP/ATP carrier-specific humoral immune responses and the cytokine-dependent anti-ADP/ATP carrier antibody IgG subclasses were examined in a cohort of ADP/ATP carrier-immunized BALB/c mice treated with anti-CD4 monoclonal antibody. Eighteen male BALB/c mice (6-8 weeks old) were randomized into 3 groups: dilated cardiomyopathy (DCM) group, DCM-tolerance (Tol) group and control group. The mice in DCM group were immunized with the peptides derived from human ADP/ATP carrier protein for 6 months and mice in the control group were sham-immunized, while the mice in DCM-Tol group were immunized with ADP/ATP carrier protein and anti-CD4 McAb simultaneously. Serum autoantibody against ADP/ATP carrier and IgG subclasses were measured by ELISA, intracellular cytokines IFN-gamma and IL-4 of Th cells were monitored with flow cytometry, and splenic T cell cytokines IFN-gamma, IL-2, IL-4 and IL-6 were detected by using real-time fluorescent quantitative PCR. The results showed that the autoantibody against ADP/ATP carrier was found in all mice in DCM group, and the antibody level, serum IgG1 and IgG2a subclasses, cytokines in T cells and Th cells were all elevated in DCM group, as compared with those in control group (P<0.01). On the other hand, in DCM-Tol group, the autoantibody level and contents of all the cytokines were significantly different from those in DCM group (P<0.01), and were close to those in control group. And the levels of IgG1, IgG2a, IgG2b and IgG3 were influenced, to varying degrees, by anti-CD4 McAb as compared with those in DCM group. All these four types of IgG subclasses were substantially decreased in DCM-Tol group as compared with DCM group. It is concluded that the treatment with anti-CD4 McAb could prevent the activation of T cells, reverse the abnormal secretion of cytokines and the imbalance between Th1/Th2 cell subsets and abnormal production of autoantibody against ADP/ATP carrier, and eventually avoid myocardial injuries.

Objective:To investigate the role of Akt signaling pathway in the regulation of breast cancer bone metastasis induced pain behavior in rat.Methods:Model rats were randomly divided into three groups:Model group,Model+GSK690693 group and Model+ Saline group.On PID 13,14 and 21,Model+GSK690693 group rats were intrathecally injected with GSK690693,a specific inhibitor of Akt.Model+Saline group were injected with saline instead.The pain related behaviors were respectively recorded on PID 0,7,14 and 21.The expression ofp-Akt in DRG used for western bloting were examed on PID 21.Results:After the injection of Akt specific inhibitor GSK690693 by intrathecal,In the Model+GSK690693 group,the threshold value of mechanical contraction reflex was increased,the spontaneous pain behavior and the expression of p-Akt in DRG decreased.On PID 14 d and 21 d,the pain behavior of rats in Model+GSK690693 group was significantly different from that of Model+Saline group and Model group (P＜0.01);On PID 21 d,There was significant statistical significance (P＜0.01) on the expression of p-Akt and Model in the ipsilateral Model+GSK690693 of DRG group,which was statistically significant (P＜0.05) compared with the Model+Saline group.Conclusion:Intrathecal injection of Akt specific inhibitor GSK690693 inhibits rat pain related behaviors induced by bone metastasis in rat breast cancer.

Objective To establish a method for detection of aminophylline in blood samples of preterm infants . Methods A molecularly imprinted electrochemical sensing film on the glassy carbon electrode surface was prepared by electropolymerization using aminophylline as the template molecule and pyrrole as the functional monomer in 0.2 mol/L HAc-NaAc buffer solution ( pH 4.0).The surface morphology and properties of molecularly imprinted sensing films were characterized by three dimensional laser scanning microscopy , differential pulse voltammetry ( DPV) and electrochemical impedance spectroscopy ( EIS) while the effects of scanning cycle number and incubation time were investigated by square wave voltammetry(SWV) method in 5 mmol/L K3[Fe(CN)6] -0.1 mol/L KCl solution.Results Under optimized experimental conditions ,the SWV peak current difference was linear to the negative logarithm of aminophylline concentration in the range from 1.0 ×10 -7 to 1.0 ×10 -3mol/L with a detection limit (S/N=3) of 0.5 ×10 -8mol/L.The recovery rate was 92.2% -101.4%.Also, the molecularly imprinted electrochemical sensor for aminophylline had good selectivity , stability and reproducibility .Conclusion The molecularly imprinted electrochemical sensor for aminophylline can be used for rapid and accurate detection of clinical blood concentrations of aminophylline molecules in preterm infants in the future .

AIM: To explore the molecular mechanism in the pathogenesis of dilated cardiomyopathy (DCM) by analyzing the expression of T cell signaling molecules in mice with autoimmune DCM. METHODS: Mouse DCM model was induced by immunizing the animals with adenine nucleotide translocase (ANT) synthetic peptides. P56lck in T cells was detected with real-time fluorescent quantitative PCR in both DCM-group and the sham-immunized controls. At the same time, flow cytometry was used for quantity of Th cell intracellular cytokine IFN-? and IL-4, ELISA for examining the level of serum anti-ANT antibody, immune histochemistry for investigating the expression of CD45 in Th cells. RESULTS: The mRNA expression of P56lck ( 1 369.51 ?874.05 vs 47.93?10.21, P

Objective To investigate the antibacterial effect of grape peel residue procyanidins on common clinical bacteria ,inclu‐ding E .coli ,Pseudomonas aeruginosa ,Acinetobacter baumannii ,Enterococcus and methicillin‐resistant Staphylococcus aureus (M R‐SA) .Methods By the means of agar plate method ,we detected the minimum inhibitory concentration (MIC) of 30 strains of E .co‐li ,28 strains of Pseudomonas aeruginosa ,15 strains of Acinetobacter baumannii ,32 strains of Feces Enterococcus ,25 strains of E . faecalis and 70 strains of MRSA ,then calculate the MIC50 and MIC90 .The result was analyzed with SPSS software 16 .0 .Results The concentration of procyanidins at 3 -8 mg/mL had no inhibitory effect on E .coli ,Pseudomonas aeruginosa and Acinetobacter baumannii;there was no inhibitory effect on E .faecalis at 3-8 mg/mL ,but the inhibition rate to Feces Enterococcus was 6 .3% .By measuring the inhibitory effect of procyanidins on 70 MRSA ,the inhibition rate at 4 mg/mL was 65 .7% ,at 8 mg/mL was 95 .7% , MIC50 was 4 .221 mg/mL and MIC90 was 6 .260 mg/mL .Conclusion There are no inhibitory effects of grape peel residue procya‐nidins on Gram‐negative bacilli such as E .coli ,Pseudomonas aeruginosa ,Acinetobacter baumannii ,and there are certain inhibitory effects on Gram‐positive bacteria such as MRSA ,enterococci ,especially on MRSA (P<0 .05) .