1.A simple microfluidic chip technology for assaying electrotaxis of cancer cells
Yonghua MI ; Wenhai XIAO ; Juan LIAO ; Jing CHEN ; Yuan LI
Military Medical Sciences 2016;40(12):946-951,957
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 .
2.Cytokine production in mice with experimental cardiomyopathy treated with anti-L3T4 monoclonal antibody at different stages
Zhaohui WANG ; Yuhua LIAO ; Jing YUAN ; Min WANG ; Jinghui ZHANG ; Yuan TIAN ; Jihua DONG
Chinese Journal of Pathophysiology 1989;0(06):-
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.
3.Comparison and application of three visual rating scales for white matter lesions in Alzheimer's disease and mild cognitive impairment
Wanxin MA ; Shan SHI ; Jing LIAO ; Huali WANG ; Xin YU ; Huishu YUAN
Chinese Journal of Geriatrics 2013;32(10):1042-1046
Objective To compare the clinical value of three visual rating scales (VRS) for white matter lesions (WML) in Alzheimer's disease (AD) and mild cognitive impairment (MCI).Methods Totally 184 subjects (including 107 AD patients,47 MCI patients and 30 normal controls)were recruited.All subjects underwent comprehensive neuropsychological tests and were examined with a standard protocol of MR imaging.WML burden was rated with the Age-Related White Matter Changes (ARWMC) rating scale,Cholinergic Pathways Hyperintensities Scale (CHIPS) and Fazekas scale.Consistence of the three rating scales were analyzed,and detection results of WML in AD/ MCI/NC group with the three rating scales were compared.The relationship between WML and cognitive function in AD and MCI groups were explored.Results ARWMC rating scale,CHIPS and Fazekas scales were moderately to highly correlated with WML (r=0.61-0.78,all P<0.01).ARWMC showed that WML were in the left and right frontal lobes in AD and MCI groups and in the right frontal lobe and left temporal lobe in NC group,which had significant differences (F=4.20,4.69,4.69,3.36,respectively,all P<0.05).WML in the left temporal lobe had a significant difference between MCI group and NC group (F=3.36,P<0.05).CHIPS showed that WML in the front left side of centrum semiovale in AD and MCI groups had significant difference as compared with that in NC group (F=4.88,P<0.05),and WML in the rear right side of low external capsule had a significant difference between AD and NC groups (F=3.04,P<0.05).Fazekas scale showed that there was no difference in WML among AD,MCI and NC groups.ARWMC showed that scores of WML in right temporal lobe were positively correlated with concentration and visual construction,and bilateral basal ganglia were negatively correlated with the concentration.Fazekas scale showed that scores of WML in periventricular were negatively correlated with concentration,MMSE and orientation,scores of WML in deep white matter were negatively correlated with concentration,abstraction and judgment,and the whole scores were negatively correlated with concentration.CHIPS showed that the scores of WML in the rear left side of low external capsule were negatively correlated with short-term memory in MCI group.Conclusions The three rating scales well consistent with each other.ARWMC rating scale and CHIPS can detect the differences in WML between AD,MCI and NC groups.The scores of CHIPS in WML are related with cognition in MCI group,while scores of ARWMC and Fazekas scale in WML are correlated with many aspects in cognitive function in AD group.In general speaking,CHIPS in MCI patients and ARWMC rating scale in AD patients show more advantages in the detection of WML and have better correlation with cognitive function.
4.Expression and significance of IL-10 mRNA in children patients with mycoplasma pneumoniae pneumonia
Wenjiang LIAO ; Tingbiao HUANG ; Anhui YUAN ; Yan MENG ; Xia FU ; Jing HE
Chinese Journal of Primary Medicine and Pharmacy 2011;18(3):289-290
Objective To investigate the expression and significance of interkeulin 10 (IL-10)in children patients with mycoplasma pneumoniae pneumonia(MP). Methods Twenty-six children patients with MP( MP group)and20 health control( contrl group)were sequentially enrolled in this study. The production of IL-10 mRNA was measured by fluorescent quantitative polymerase chain reaction. Results Expression of IL-10 mRNA in MP group (33 893.22±5 207.60) copy/μg was significantly lower than that in control group(59 341.92 ±6 712.98) copy/μg(t = - 14. 49, P < 0. 01 ). Expression of IL-10 mRNA in heavy degree MP group was significantly lower than that of mild degree MP group ( t= - 2. 20, P < 0. 05 ). Conclusion The expression of IL-10 was low, and played an important role in the development of patients with MP.
5.Multi-modality magnetic resonance imaging features of cognitive function in mild cognitive impairment
Huali WANG ; Huishu YUAN ; Yue ZHU ; Jing LIAO ; Meiyan ZHANG ; Tao LI ; Xin YU
Chinese Journal of Internal Medicine 2010;49(8):680-683
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.
6.MR diffusion tensor imaging-based white matter studies in mild cognitive impairment and Alzheimer disease
Jing LIAO ; Huishu YUAN ; Yue ZHU ; Meiyan ZHANG ; Xin YU ; Huali WANG
Chinese Journal of Radiology 2009;43(5):490-494
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.
7.Antibacterial effect of grape peel procyani residudins on common clinical bacteria
Li FANG ; Jing WANG ; Yuan XU ; Yan CAI ; Tao LIAO ; Zhong TANG
Chongqing Medicine 2015;(4):453-455
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) .
8.Research on the Inhibition of Breast Cancer Bone Metastasis Induced Rat Pain-related Behaviors by Intrathecally Injection of GSK690693, an Akt Specific Inhibitor
Zhijun LIAO ; Yunlong ZHOU ; Jing LI ; Hongxiang LIANG ; Cuitang YUAN ; Gang DING
Progress in Modern Biomedicine 2017;17(25):4812-4815,4823
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.
9.Analysis of specific Th1/Th2 helper cell responses and IgG subtype antibodies in anti-CD4 monoclonal antibody treated mice with autoimmune cardiomyopathy.
Zhaohui, WANG ; Yuhua, LIAO ; Jing, YUAN ; Jinghui, ZHANG ; Jihua, DONG ; Jinping, WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2008;28(4):409-14
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.
10.Preparation and application of aminophyllin electrochemical sensor based on molecularly imprinting technique
Jing CHEN ; Wenhai XIAO ; Juan LIAO ; Qingmei HE ; Yuan LI ; Fang GONG
Military Medical Sciences 2016;40(12):952-957
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 .