Objective:To analyze the effects of silver ion on the integration frequency of the class 1 integron in Escherichia coli ( E. coli) BL21(DE3) host. Methods:Two recombinant plasmids, pUCINT and pACINAD, were successively transformed into E. coli BL21 (DE3) to construct HS2 strains. Three experimental groups were set up using 0.3 μg/ml, 0.6 μg/ml and 0.8 μg/ml silver ion LB liquid medium, while control group used common LB liquid medium. Silver ion was supplied by silver nitrate and HS2 strains were cultured at 37℃ for 24 h. The copy number of cointegrates and the total copy number of integrons in each group were detected by real-time polymerase chain reaction (qPCR), and the ratio of them was the integration frequency. Changes in the integration frequency were analyzed by three independent phenotypic screening method and the protein expression in HS2 strains was analyzed by mass spectrometry. Results:The integration frequency in HS2 strains in the control group and three experimental groups (0.3 μg/ml, 0.6 μg/ml and 0.8 μg/ml silver ion) was 1.79×10 -5 (1.44×10 -5, 3.13×10 -5), 2.07×10 -5 (1.49×10 -5, 2.67×10 -5), 2.25×10 -6 (1.47×10 -6, 4.54×10 -6) and 1.69×10 -6 (0.22×10 -6, 3.08×10 -6), respectively. The integration frequency in the 0.6 μg/ml and 0.8 μg/ml silver ion groups was significantly lower than that in the control group ( P<0.01), but there was no significant difference between the 0.3 μg/ml silver ion group and the control group. Results of three independent phenotypic screening method were consistent with those obtained by qPCR. Mass spectrometry analysis showed that there were differences in protein expression in HS2 strains between the control group and the experimental groups. Conclusions:Silver ion at a certain concentration had an inhibitory effect on the frequency of drug resistance gene cassette captured by bacterial integron.

BACKGROUND:Diabetic cystopathy is one of the most common chronic diabetic complications. The establishment of animal models of diabetic cystopathy wil provide experimental animal platform for relevant research.
OBJECTIVE:To establish a guinea pig model of diabetic cystopathy and to evaluate its urodynamic characteristics.
METHODS:Fifty short-hair Britain female guinea pigs were randomly divided into two groups, 42 as the experiment group and the other 8 as the control group. The experiment group was intraperitoneal y injected with streptozotocin to induce diabetes. The control group received injection of blank citric acid buffered solution. Diabetic guinea pigs were detected by urinary dynamics test at 9 and 12 weeks. Diabetic guinea pigs were further assigned into diabetic cystopathy subgroup and compensated subgroup. The urodynamic parameters of three groups were compared.
RESULTS AND CONCLUSION:Twenty of 42 guinea pigs were successful y induced diabetes by the injection of streptozotocin. At 9 weeks after the injection, bladder function compensation was present in six diabetic guinea pigs while bladder function was decompensated in another three diabetic guinea pigs. At 12 weeks, bladder function compensation was present in one diabetic guinea pig, while another eight guinea pigs were confirmed with diabetic cystopathy (88.89%). In the diabetic cystopathy subgroup, the residual urine volume was increased (0.72±0.08) mL, maximal detrusor pressure was decreased (0.63±0.05) kPa, maximum bladder capacity was increased (2.01±0.05) mL, and bladder compliance was increased (0.34±0.04) mL/kPa. There were significant differences compared with the compensated subgroup and the control group (P<0.001). Diabetic cystopathy occurs at 12 weeks after diabetic models are successful y established in guinea pigs, and urodynamic changes are mainly the increase of residual urine volume.

Objective To evaluate the effects of amiodarone on connexin43 (Cx43) expression during myocardial ischemia-reperfusion (I/R) in rats.Methods Adult male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 3 groups (n =12 each) using a random number table:shame operation group (group S),I/R group and amiodarone group (group AM).Myocardial ischemia was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 120 min of reperfusion in I/R and AM groups.Amiodarone 5.0 mg/kg was injected intravenously at 10 min before ischemia followed by infusion at a rate of 0.15 mg· kg-1 · min 1 until the end of reperfusion.Arrhythmia was recorded during reperfusion and scored.At the end of reperfusion,blood samples were taken from the femoral artery for determination of the serum levels of cardiac troponin Ⅰ (cTnI) and creatine kinase isoenzyme-MB (CK-MB).Then the animals were sacrificed and myocardial specimens were removed for determination of the expression of Cx43 protein (by immuno-histochemistry) and mRNA (by RT-PCR) in myocardial tissues.Results Compared with group S,the arrhythmia score and serum levels of CK-MB and cTnl were significantly increased,and the expression of Cx43 protein and mRNA was downregulated in I/R and AM groups (P ＜ 0.05).Compared with group I/R,the arrhythmia score and serum levels of CK-MB and cTnI were significantly decreased,and the expression of Cx43 protein and mRNA was up-regulated in group AM (P ＜ 0.05).Myocardial Cx43 was unevenly distributed in group I/R,while evenly distributed in S and AM groups.Conclusion The mechanism by which amiodarone protects myocardium against I/R-induced arrhythmia is related to inhibition of redistribution of Cx43 and up-regulation of Cx43 expression in rats.

This paper introduced a hand function rehabilitation system based on motor imagery (MI) brain-computer interface for hand function rehabilitation of stroke patients. The rehabilitation system contains three subsystems. Offline training subsystem displays the blank screen, a left or right hand movement video and arrow in turn, which respectively reminders patients to rest and make preparations for MI and instruct them how to do MI, and be doing MI. Finally, the patients' electroephalography (EEG) signals are acquired and processed togenerate a recognition model. Model update online training subsystem presents the black screen and a left or right arrow, the meanings ofwhich are the same as those in offline training subsystem. Then the acquired EEG signals are analyzed according to the established recognitionmodel. Next, the analysis result is derived to control the hand movement video to be played. The video can also act as a visual feedback,which makes patients' EEG signals easier to be recognized. The updated and more effective recognition model is built at last. Virtual reality(VR) online training subsystem constructs 3D grid models of VR scene, a 3D man model and its hand animations in the 3Dmax. Then, all ofthem are imported into Unity3D. The control methods of the animations are also designed in Unity3D. In the end, the patients' EEG signalsare analyzed according to the updated recognition model, thus controlling the hand movements of the 3D man in real time. The developedsystem has many characteristics, such as multilevel training and more immersion, which hopefully promotes the plasticity of central nervoussystem. The designed system provides new treatments for post-stroke hand function rehabilitation and further lays the foundation for family-mode rehabilitation.

Objective To investigate the relationship between promoter hypermethylation of secreted frizzled-related protein (SFRP) gene and acute leukemia (AL). Methods We examined the promoter methylation starus of SFRP1, 2, 4 and 5 in primary or relapsed AL patients, cell lines ( HL60,NB4, Molt-4 and Jurkat) and peripheral blood mononuclear cells from healthy people with methylationspecific PCR (MSP). Results None of the normal mononuclear cells showed methylation of any SFRP genes. The frequencies of aberrant methylation among the samples were 33.9% (20/59) for SFRP1,23.7%(14/59) for SFRP2, 6. 8% (4/59) for SFRP4 and 10. 2% (6/59) for SFRP5 in acute myelocytic leukemia (AML), and 39.3% (11/28) for SFRP1, 28.6% (8/28) for SFRP2, 25.0% (7/28) for SFRP4 and 32. 1% (9/28) for SFRP5 in acute lymphoblastic leukemia (ALL). Hypermethylation of SFRP1, 2, 5genes were present in the 4 AL cell lines. SFRP4 was methylated in NB4, Molt-4 and Jurkat cell lines.However, methylation and unmethylation of SFRP4 were both detected in HL60. Conclusions Hypermethylation of SFRP genes is a common event in the evolution of AL. Methylation of SFRP genes might serve as potential independent biomarkers for early detection of AL.

Objective The study was to compare the effect of sufentanil and morphine preconditioning on ischemia /reperfusion-induced ventricular arrhythmias and the expression and distribution of myocardial Cx43 in rats.The regulation mechanisms that how sufentail and morphine lead to the change of Cx43 were also studied.Methods 32 male SD rats were randomly divided into 4 groups:sham operation group (group C),ischemia/reperfusion (group I/R),morphine preconditioning group(group M) and sufentanil preconditioning group(group S),each group had 8 rats.Established myocardial ischemia/reperfusion model,continuous recorded Ⅱ ECG,mean arterial pressure(MAP) and heart rate(HR).The ventricular arrhythmias at the 30 min before reperfusion was observed and the ventricular arrhythmias score of each group was calculated by ECG analysis; expression and distribution of Cx43 protein were observed by immunohistochemical technique.Results Compared with group C,the HR,MAP,RPP of group I/R were decreased obviously (P ＜ 0.05),while the arrhythmia score was significantly higher(P ＜ 0.05).Compared with group I/R,the extent of the declined of HR,MAP,RPP of group M and group S were eased significantly(P ＜ 0.05) and arrhythmia score was significantly lower(P ＜ 0.05).The HR,MAP,RPP of group M and group S are closer(P ＞ 0.05).Compared with the group C,Cx43 expression level in group I/R was significantly reduced (P ＜ 0.05) and the distribution was disordered,while compared with the group I/R,Cx43 expression level in group M and group S were significantly increased (P ＜ 0.05),and its distribution was structured.In group M and group S,Cx43 expression level were closer(P ＞ 0.05) and so as their distribution.Conclusion Sufentanil and morphine could inhibit the reduction of myocardial Cx43 expression level and improve its distribution which could played an important role in anti-arrhythmic during ischemia-reperfusion.

Objective To study the radiation injury of rat C6 glioma cell line by high resolution,1 H-nuclear magnetic resonance (1 H NMR) spectroscopy,and to preliminarily investigate its mechanism.Methods Metabolite concentrations in C6 cells were determined by 1 H NMR spectroscopy.Comet assay was used to evaluate DNA damage.Flow cytometry was used to determine the cell cycle and apoptosis rate.Colony-forming assay was used to measure the colony-forming rate and preliminarily investigate the mechanism of radiation injury.The resuhs were analyzed by one-way analysis of variance and Pearson correlation analysis.Results With the increase in radiation dose from 0 Gy to 1,5,10,and 15 Gy,DNA damage was enhanced in a dose-dependent manner (P=0.000-0.690);the percentage of cells in G1 phase increased (P =0.026-0.749);the apoptosis rate significantly increased (all P =0.000);the colony-forming rate significantly declined (P =0.000-0.004);the Lac/Cr ratio significantly decreased (P =0.000-0.015),which had a negative linear correlation with DNA damage parameters (tail length,r=-0.971;％DNA in the tail,r =-0.998;tail moment,r =-0.995) and apoptosis rate (r =0.978).Conclusions 1 H NMR spectroscopy reveals that the change in the Lac/Cr ratio is associated with injury and apoptosis of C6 cells after radiation.1 H NMR spectroscopy has the potential to predict radiation injury of glioma.

Objective To investigate the inhibition of proliferation and the regulation of histone acetylation modification in Jurkat cells treated by sodium valproate(VPA). Methods Jurkat cells were treated with VPA.Cell proliferation was determined by CCK-8 assay, and cell cycle were analyzed by flow cytometry (FCM). mRNA of HDAC1 was detected by semi-quantitative RT-PCR, and protein expression of HDAC1 and acetylation of histone H3, H4 was examined by Western blotting. Results VPA inhibited the proliferation of Jurkat cells in concentration-and time-dependent manners. After exposure to VPA in different concentrations for 48h,cell cycle was arrested obviously at G0/G1 phase (P <0.05), and with increasing concentration, the percentage of G0/G1 phase cells was increased and that of S phase were decreased. HDAC1 mRNA expression were inhibited with the increasing concentration of VPA. The protein level of HDAC1 was down-regulated, while acetylation of histone H3、H4 was up-regulated in Jurkat cells by VPA. Conclusion VPA can inhibit proliferation of Jurkat cells and induce G0/G1 phase arrest. The mechanism may be that VPA increase acetylation of histone H3/H4 by inhibiting expressions of HDAC1 gene.

Objective To evaluate the predictive effect of Braden scale for the risk of development of pressure ulcers (PU) in the department of neurology bedridden patients and to explore subgroup preventive measures. Methods 400 cases newly hospitalized bedridden patients in the department of neurology were collected with no pressure ulcers at the first evaluation and pressure ulcer risk was continuously predicted by a Braden scale skin assessment. The high-risk, middle-risk and low-risk groups were randomized into the experimental group and the control group respectively. Routine preventive measures were taken for the control group while the air fluidized bed for the high-risk group, the sponge mattress for the middle-riskgroup, and turning the body over every 4 hours for low-risk group. Other preventive procedures were undertaken simultaneously in beth the experimental and the control groups. Results The area under the ROC curve (AUC)was 0.771 and 0.828 at the first and last time Braden scale scores respectively. Such vMues as sensitiveness, specificity, positive predictive value, negative predictive value were found in higher level,when the diagnosis value was 17. There was no significant difference of incidence rate of the subgroup pressure ulcers between the high-risk, middle-risk, low-risk groups compared to the control group. Conclusions The effect of predicting pressure ulcer risk for bedridden patients in the department of neurology with Braden scale was fairly good, while the score 17 as the diagnosis value was ideal. The air fluidized bed for the high-risk group and the sponge mattress for the middle-risk group resulted in no significant decrease of incidence rate of the pressure ulcer, while taming over the patients' body every 4 hours for low-risk groups showed acceptable and therefore saving medical resources.

GeneHub software developed by us can be applied to evaluate the significance of expression similarity of genes in each functional unit of different gene function classification systems, and thus to select experiment condition related gene functional units. With different expression similarity measurement and different experiment data, we have applied GeneHub to analyze the expression correlation of functional related genes classified respectively with gene chromosome location, celluar location and interaction, metabolic pathway and signal transduction pathway relationship of protein products of genes. Genes classified according to these multiple ways all showed significant experimental condition related co-expression in our analysis. Our results provide further evidence for the assumption that functional related genes express similarly, which is widely adopted in gene expression profile analysis.
Chromosome Mapping ; methods ; Expressed Sequence Tags ; Gene Expression Profiling ; methods ; Gene Expression Regulation ; physiology ; Humans ; Oligonucleotide Array Sequence Analysis ; methods ; Software ; User-Computer Interface