Objective To study the damage focused ultrasound inflicts on the membrane permeability of Ehrlich ascitic tumor (EAC) cells and the relationship between changes in membrane permeability and focused ultra-sound exposure time. Methods The relative survival rate of tumor cells was examined at various intensities and dif-ferent exposure times using focused 2.2 MHz ultrasound. The uhrastructure changes were evaluated with a scanning electron microscope after different exposures. Membrane permeability was investigated by incorporating fluorescein isothiocyanate dextran (FD5OO) , and membrane damage was evaluated by measuring lactate dehydrogenase (LDH) release. Results Morphological observation showed there were numerous microvilli on the surface of un-exposed cells. When the cells had been irradiated with focused ultrasound for 30 s there was only a slight effect on the shape of the cells and the number of microvilli was slightly reduced. When the cells were exposed to ultrasound for 60 s, the surface of many cells became relatively smooth with no obvious microvilli, and several small craters were seen on the surfaces of cells where the cytoplasm seemed to have extruded through the membrane. The cell membrane was seri-ously damaged by sonoporation. The loading of FD500 in the unexposed cells was only 0.21%. When the cells had been sonicated with focused ultrasound for 30 s or 60 s, the loading of FD500 increased to 11.46% and 18.50% re-spectively. The released LDH activities in the 30 s group and 60 s group were 2.94±0.02 and 3.28±0.04 U/L, respectively. The activities of LDH increaased as the focused ultrasound exposure time was prolonged. Conclusion Focused ultrasound may damage the cell membrane permeability of EAC cells, and the damage increases as the expo-sure time is prolonged from 30 s to 60 s.

Objective:To investigate the early diagnosis value of neutrophilic CD 64 index(nCD64 ID),neutrophilic CD32 index( nCD32 ID) in ascites and CRP in blood of liver cirrhosis patients combined with spontaneous bacterial peritonitis. Methods:The data of 156 cases with liver cirrhosis was analyzed retrospectively, which CD32 index, CD64 index and CRP were detected respectively and ROC curve analysis were performed. Results:The nCD64 ID,nCD32 ID and CRP in bacterial infection group were all significantly higher than that in no infection group(P<0. 001). The sensitivity and specificity of nCD32 ID,nCD64 ID and CRP were 82. 8%,96. 2%,72. 5% and 81. 0%, 95. 8%, 73. 1% respectively. Conclusion: The sensitivity and specificity of nCD64 ID were higher than nCD32 ID and CRP. The nCD64 ID can be used as an effective index for early diagnosis and differential diagnosis of liver cirrhosis combined with spontaneous bacterial peritonitis.

Objective To observe the mRNA expressions of endothelin-1(ET-1) and endothelin A/B receptors (ETA/B) in tissue of benign prostatic hyperplasia treated by daily low-dose sildenafil.Methods A total of 32 patients with benign prostatic hyperplasia were randomly divided into two groups:treatment(25 mg sildenafi for 12 weeks,n=16) and control (no drug,n=16) groups.Immunohistochemical staining,ELISA and RT-PCR were used to detect the expression levels of ET-1 protein and ET A/B mRNA,respectively.Results The expressions of ET-1 protein and ET A/B mRNA in prostatic tissue were significantly lower in treatment group than in control group[(53.31±18.56) ng/kg vs.(83.34±31.38) ng/kg,0.356±0.056 vs.0.624±0.083,0.721±0.083 vs.0.933±0.905,t=-3.295,10.715,6.937,all P＜0.001].Conclusions Daily low-dose sildenafil can reduce the expressions of ET-1 and ET A/B receptors mRNA in benign prostatic hyperplasia.

Objective To detect the level of CD64 and serum procalcitonin (PCT ) and investigate the diagnosis value of CD64 and serum PCT in cirrhosis patients with spontaneous bacterial peritonitis (SBP) .Methods Participants were categorized in‐to three groups including liver cirrhosis with SBP(45 patients) ,liver cirrhosis without SBP(93 patients) and health personnel(50 persons) .CD64 was detected by flow cytometry and serum PCT was measured by electroc hemiluminescence immunoassay .The li‐mosis vein blood samples were obtained from the patients with SBP at the time of 24 h after admission ,before antibacterial drugs use and 7 days after the effective treatment of antibacterial drugs .The CD64 and serum PCT were detected with the limosis vein blood samples .At the same time ,the complete blood count ,liver ,kidney and blood coagulate functions were tested .The participants in other two groups were detected the CD64 ,serum PCT ,complete blood count ,liver ,kidney and blood coagulate functions at the same time .Results The level of CD64 and serum PCT in cirrhosis patients with SBP were significantly higher than those in liver cirrhosis without SBP and normal controls (P< 0 .01) .ROC curve analysis showed that the sensitivity and specificity of CD64 and serum PCT were 95 .5% ,93 .8% and 96 .1% ,85 .2% respectively .Conclusion CD64 and serum PCT can be determined as the im‐portant indicator in early diagnosis and efficacy criterion .

Objective To explore the value of MRI histogram analysis in the risk assessment of medulloblastoma recur rence.Methods The data of 28 patients pathologically confirmed of medulloblastoma was analyzed retrospectively.All patients were divided into recurrent group and the non recurrent group (each n=14).The ROIs were drawn on the maximum level of enhanced MR sagittal images,and the histogram analysis were performed using the software named Mazda.The statistical analysis was performed on the histogram parameters to find out the different characteristics between the two groups,and the ROC curve was drawn to evaluate its diagnostic efficacy for recurrence of medulloblastoma.Results In all of the 9 parameters which are extracted from histogram,kurtosis had statistical significance between the 2 groups (P=0.018).The area under the ROC curve was 0.776 (P=0.018),and the sensitivity and specificity of kurtosis in the risk assessment of medulloblastoma recurrence were 64.3％ and 71.4％,respectively.Conclusion MRI histogram analysis can be an important method to assess the risk of medulloblastoma recurrence.

Objective: To investigate the effects of Paraquat on neural stem cell proliferation in vitro and explore the its mechanism based on DNA methylation pathway. Methods: Nestin, β-tubulin III, and glial fibrillary acidic protein (GFAP) were detected by indirect immunofluorescence assay to evaluate self renewal and differentiation potentia of ReNcell CX human neural stem. The cells were treated with terminal concentrations of 0, 5, 25, 50, and 100μmol/L PQ for 24 hours, and the cells were induced by 50 μmol/L PQ for different time (6, 12, 24, 48 h). Cell viability was determined by MTT assay. The proliferation of neural stem cells was evaluated using Sox2/Brdu and Nestin/Brdu double immunofluorescence staining. The global DNA methylation level was assayed by MethyflashTM methylated DNA Quantification kit. The expression levels of Dnmts mRNA and protein were analyzed by quantitative reverse transcription polymerase chain reaction(qRT-PCR) and Western blot, respectively. Results: Immunofluorescence showed that nestin was primarily expressed in proliferative neural stem cell and peotein biomarkers (β-tubulin III, GFAP) for neuron and astrocyte were expressed in differentiated cells. MTT assay showed PQ induced cell survival rate decrease in a time and dose dependent manner. Double immunfluorescence staining of cells showed colocalization of Sox2 and Brdu. The percentage of Brdu/Sox2 positive cells was significantly lower in the PQ-exposed (25, 50, 100μmol/L PQ treatment) groups compared to control (P<0.05); Meanwhile, The percentage of Brdu/Nestin positive cells was also significantly lower in the PQ-exposed(50,100μmol/L PQ treatment) groups compared to control (P<0.05). The results of global DNA methylation revealed a significant decrease in PQ-exposed groups (P<0.05). Western blot showed that compared with control group, the protein and mRNA levels of Dnmt1, Dnmt3a in PQ-exposed group were significantly decreased (P<0.05), but there was a significant increase in expression level of Dnmt3b in 50, 100 μmol/L PQ-treated group(P<0.05). Conclusion Paraquat could inhibite the proliferation of human neural stem cells through reducing the level of DNA methylation reaction by suppressing the protein expression and transcription of DNA methylated transferase(Dnmts).

Objective: To investigate the roles of p38 mitogen-activated protein kinases (p38 MAPK) , extracellular regulated protein kinases (ERK) and c-Jun N-tenninal kinases (JNK) of MAPK signaling pathway in Paraquat-induced epithelial to mesenchymal transition (EMT) of type II alveolarepithelial cells. Methods: RLE-6NT cells were incubated with different concentrations of PQ (0, 25, 50, 100μmol/L) for 6, 12 and 24 h. Cell morphology alteration was observed under phase-contrast microscopy. Cell viability was determined using an MTT assay. Cell migration ability was detected using scratch wound assay. Protein expression of P-p38 MAP, P-Erk1/2, P-JNK, E-cad, ZO-1, Vimentin and а-SMA were detected by western blot. The level of genes related to fibrosis (COL-I, COL-III, FN and FSP-1) were analyzed via quantitative real-time RT-PCR. Results: Cell morphology started to undergo EMT changes with a phenotype characteristic of mesenchymal cells, including an elongated shape and a lack of tight cell-cell adhesions induced by 100μmol/L PQ treatment in a time-dependent manner. MTT showed that cell viability decreased with increasing PQ concentration (50、100、200、300 μmol/L PQ treatment for 24 h) and increasing treatment time (200 μmol/L PQ treatment for 6, 12, 24, 36, 48 h) . Compared to control group, the expressions of the epithelial phenotype marker E-cad and ZO-1 significantly decreased with PQ treatment (50, 100μmol/L) in a time-dependent manner (P<0.05) . Additionally, the level of the mesenchymal marker (a-SMA, vimentin) dramatically increased with PQ treatment in the same concentration-and time-dependent manner (P<0.05) . Cell migration ability was markedly increased after 24 h of 100 μmol/L PQ treatment compared to control (P<0.05) . The phosphorylated forms of p38 MAPK, Erk1/2, and JNK were increased at 24 h after stimulation with PQ (P<0.05) . This PQ induced (100 μmol/L) phosphorylation was markedly attenuated in the presence of the p38 MAPK, ERK and JNK inhibitors (SB-203580, SP-600125 and PD98059) respectively. Furthermore, RT-PCR showed that PQ significantly induced the upregulation expression of COL I and III mRNA, Fn, and FSP-1 mRNA (P<0.05) . Conclusion PQ-induced pulmonary fibrosis occurs via EMT, which is mediated by the MAPK pathway.

Humans ; Cardiac Resynchronization Therapy ; Cardiac Pacing, Artificial ; Heart Ventricles ; Heart Failure/therapy* ; Treatment Outcome ; Electrocardiography ; Ventricular Function, Left

Objective To analyze human gait stability by acceleration signal at the head and lumbar under different walking conditions, and make comparison with parameters by the traditional COM (center of mass)-COP(center of pressure) method, so as to discuss the reliability of applying wearable sensors to analyze human gait stability. Methods The harmonic ratio (HR) parameter at the head and lumbar based on acceleration signal was applied to analyze gait stability of 18 healthy young adults under 3 walking conditions (footwear normal walking, barefoot normal walking and barefoot walking at different velocities), and the results were compared with the assessment results from the COM-COP method. Results Walking at normal velocity was most stable, with the maximum HR parameter. Compared with footwear walking, HR parameters were significantly decreased (P＜0.05) during barefoot walking, indicating that gait stability was reduced. The results were consistent with the assessment results from the COM-COP method. Considering the factors of walking velocity and footwear, the gait stability parameters obtained by the two methods showed a significant negative linear correlation (R2＞0.50). Lumbar HR parameter and COM-COP parameters showed a stronger linear correlation (R2＞0.65). Conclusions The application of acceleration signal-based analysis algorithm could effectively and reliably evaluate the stability of human gait, and acceleration at the lumbar was more sensitive than the head signal for analyzing gait stability.