Wavelet entropy is a quantitative index to describe the complexity of signals. Continuous wavelet transform method was employed to analyze the spontaneous electroencephalogram (EEG) signals of mild, moderate and severe Alzheimer's disease (AD) patients and normal elderly control people in this study. Wavelet power spectrums of EEG signals were calculated based on wavelet coefficients. Wavelet entropies of mild, moderate and severe AD patients were compared with those of normal controls. The correlation analysis between wavelet entropy and MMSE score was carried out. There existed significant difference on wavelet entropy among mild, moderate, severe AD patients and normal controls (P<0.01). Group comparisons showed that wavelet entropy for mild, moderate, severe AD patients was significantly lower than that for normal controls, which was related to the narrow distribution of their wavelet power spectrums. The statistical difference was significant (P<0.05). Further studies showed that the wavelet entropy of EEG and the MMSE score were significantly correlated (r=0.601-0.799, P<0.01). Wavelet entropy is a quantitative indicator describing the complexity of EEG signals. Wavelet entropy is likely to be an electrophysiological index for AD diagnosis and severity assessment.
Mass point-spring model is one of the commonly used models in virtual surgery. However, its model parameters have no clear physical meaning, and it is hard to set the parameter conveniently. We, therefore, proposed a method based on genetic algorithm to determine the mass-spring model parameters. Computer-aided tomography (CAT) data were used to determine the mass value of the particle, and stiffness and damping coefficient were obtained by genetic algorithm. We used the difference between the reference deformation and virtual deformation as the fitness function to get the approximate optimal solution of the model parameters. Experimental results showed that this method could obtain an approximate optimal solution of spring parameters with lower cost, and could accurately reproduce the effect of the actual deformation model as well.
ObjectiveTo evaluate the combination of lipopolysaccharide-amine nanopolymersomes (LNPs), as a gene vector, with target gene and the transfection in bone marrow mesenchymal stem cells (BMSCs) so as to provide a preliminary experiment basis for combination treatment of bone defect with gene therapy mediated by LNPs and stem cells. MethodsPlasmid of bone morphogenetic protein 2 (pBMP-2)-loaded LNPs (pLNPs) were prepared. The binding ability of pLNPs to pBMP-2 was evaluated by a gel retardation experiment with different ratios of nitrogen to phosphorus elements (N/P). The morphology of pLNPs (N/P=60) was observed under transmission electron microscope (TEM) and atomic force microscope (AFM). The size and Zeta potential were measured by dynamic light scattering (DLS). The resistance of pLNPs against DNase I degradation over time was explored. The viability of BMSCs, transfection efficiency, and expression of target protein were investigated after transfection by pLNPs in vitro. ResultsAt N/P≥1.5, pLNPs could completely retard pBMP-2; at N/P of 60, pLNPs was uniform vesicular shape under AFM; TEM observation demonstrated that pLNPs were spherical nano-vesicles with the diameter of (72.07±11.03) nm, DLS observation showed that the size of pLNPs was (123±6) nm and Zeta potential was 20 mV; pLNPs could completely resist DNase I degradation within 4 hours, and such protection capacity to pBMP-2 decreased slightly at 6 hours. The cell survival rate first increased and then decreased with the increase of N/P, and reached the maximum value at N/P of 45; the cytotoxicity was in grade I at N/P≤90, which meant no toxicity for in vivo experiment. While the transfection efficiency of pLNPs increased with the increase of N/P, and reached the maximum value at N/P of 60. So it is comprehensively determined that the best N/P was 60. At 4 days, transfected BMSCs expressed BMP-2 continuously at a relatively high level at N/P of 60. ConclusionLNPs can compress pBMP-2 effectively to form the nanovesicles complex, which protects the target gene against enzymolysis. LNPs has higher transfection efficiency and produces more amount of protein than polyethylenimine 25k and Lipofectamine 2000.
Attention-deficit/hyperactivity disorder (ADHD) is one of the most common neuro-developmental disorders occurring in childhood, characterized by symptoms of age-inappropriate inattention, hyperactivity/impulsivity, and the prevalence is higher in boys. Although gray matter volume deficits have been frequently reported for ADHD children via structural magnetic resonance imaging, few of them had specifically focused on male patients. The present study aimed to explore the alterations of gray matter volumes in medicated-naive boys with ADHD via a relatively new voxel-based morphometry technique. According to the criteria of DSM-IV-TR, 43 medicated-naive ADHD boys and 44 age-matched healthy boys were recruited. The magnetic resonance image (MRI) scan was performed via a 3T MRI system with three-dimensional (3D) spoiled gradient recalled echo (SPGR) sequence. Voxel-based morphometry with diffeomorphic anatomical registration through exponentiated lie algebra in SPM8 was used to preprocess the 3D T1-weighted images. To identify gray matter volume differences between the ADHD and the controls, voxel-based analysis of whole brain gray matter volumes between two groups were done via two sample t-test in SPM8 with age as covariate, threshold at P＜0.001. Finally, compared to the controls, significantly reduced gray matter volumes were identified in the right orbitofrontal cortex (peak coordinates [-2,52,-25], t=4.01), and bilateral hippocampus (Left: peak coordinates [14,0,-18], t=3.61; Right: peak coordinates [-14,15,-28], t=3.64) of ADHD boys. Our results demonstrated obvious reduction of whole brain gray matter volumes in right orbitofrontal cortex and bilateral hippocampus in boys with ADHD. This suggests that the abnormalities of prefrontal-hippocam-pus circuit may be the underlying cause of the cognitive dysfunction and abnormal behavioral inhibition in medicated-naive boys with ADHD.
ObjectiveTo investigate the long-term effectiveness of microgenia treatment with natural coral, and the volume relationship between the implant and the new bone. MethodsA retrospective analysis was made on the cl inical data of 12 patients with microgenia treated by horizontal genioplasty with natural coral implantation between October 1998 and September 2004. There were 7 males and 5 females with the average age of 18.5 years (range, 15-28 years). The cephalometric data on the photo and X-ray films were collected at pre-operation, immediate after operation, and last follow-up. The vertical distance between lower lip point and inferior mental point, the vertical distance between inferior alveolar point and inferior mental point, the vertical distance of the osteotomic gap, and the distance between pogonion and the line between nasion and inferior alveolar point were measured, and the recurrence rates were caculated. ResultsAll incisions healed by first intention, and no complication occurred. All patients were followed up 8-12 years (mean, 9.2 years). X-ray films showed that the natural coral was replaced by new bone formation in the mental osteotomic gap; the new bone had good strength and firmly attached to the mentalis and periosteum. At last follow-up, the vertical distance between lower lip point and inferior mental point, the vertical distance between inferior alveolar point and inferior mental point, and the vertical distance of the osteotomic gap were decreased when compared with the ones at immediate after operation, and the mean recurrence rates were 6.1%, 22.9%, and 31.7%, respectively; and no obvious change was observed in the vertical distance between pogonion and the line between nasion and inferior alveolar point. Nine patients were satisfied with operation effectiveness; chin morphology was adjusted again in 3 patients. ConclusionNatural coral is a safe and effective bone substitute with enough stable new bone and good long-term effectiveness.
ObjectiveTo explore the function of intercellular adhesion A (icaA), fibrinogen binding protein (fbe), and accumulation-associated protein (aap) genes in formation of Staphylococcus epidermidis-Candida albicans mixed species biofilms. MethodsThe experiment was divided into 3 groups:single culture of Staphylococcus epidermidis ATCC35984 (S. epidermidis group) or Candida albicans ATCC10231 (C. albicans group), and co-culture of two strains (mixed group) to build in vitro biofilm model. Biofilm mass was detected by crystal violet semi-quantitative adherence assay at 2, 4, 6, 8, 12, 24, 48, and 72 hours after incubation. XTT assay was performed to determine the growth kinetics in the same time. Scanning electron microscopy (SEM) was used to observe the ultrastructure of the biofilms after 24 and 72 hours of incubation. The expressions of icaA, fbe, and aap genes were analyzed by real-time fluorescent quantitative PCR. ResultsCrystal violet semi-quantitative adherence assay showed that the biofilms thickened at 12 hours in the S. epidermidis and mixed groups; after co-cultured for 72 hours the thickness of biofilm in mixed group was more than that in the S. epidermidis group, and there was significant difference between 2 groups at the other time (P<0.05) except at 72 hours (P>0.05). In C. albicans group, the biofilm started to grow at 12 hours of cultivation, but the thickness of the biofilm was significantly lower than that in the mixed group in all the time points (P<0.05). XTT assay showed that the overall growth speed in the mixed group was greater than that in the C. albicans group, and it was greater than that in the S. epidermidis group at 48 hours; there was no significant difference in the growth speed between the mixed groups and the S. epidermidis group in the other time points (P>0.05) except at 12 hours (P<0.05). The absorbance (A) value in the mixed group was lower than that in the S. epidermidis group at 2 and 4 hours, but no significant difference was shown (P>0.05); the A value of mixed group was significantly higher than that of the C. albicans group after 6 hours (P<0.05). SEM observation showed that mature biofilms with complex structure formed in all groups. The real-time fluorescent quantitative PCR showed the expressions of fbe, icaA, and aap genes in mixed group increased 1.93, 1.52, and 1.46 times respectively at 72 hours compared with the S. epidermidis group (P<0.05). ConclusionMixed species biofilms have more complex structure and are thicker than single species biofilms of Staphylococcus epidermidis or Candida albicans, which is related to increased expressions of the icaA, fbe, and aap genes of Staphylococcus epidermidis.
ObjectiveTo establish an in vitro model of Candida albicans-Staphylococcus epidermidis mixed species biofilm on polyvinyl chloride (PVC) material, and to observe mixed species biofilm formation and its microstructure. MethodsStaphylococcus epidermidis bacteria (ATCC35984) and Candida albicans fungal (ATCC10231)were co-incubated with 0.5 cm diameter PVC pieces in tryptic soy broth (TSB) to form mixed specie biofilms (experimental group). At 2, 6, 12, 24, 48, and 72 hours, the thicknesses of the biofilms, the number of bacteria per sight, and the percentage of viable cells in biofilms were measured, and three-dimensional images of biofilms were obtained using confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) at 48 hours. PVC material cultured in the TSB medium served as control group. ResultsIn control group, there was no pathogenic bacteria adhesion on the PVC material surface. In experimental group, CLSM showed that colonies and biofilm formation were found at 6 hours after co-culture, and gradually increased with time. The pathogenic bacteria colonies reached the peak at 24 hours, and biofilm thickness attained peak value at 48 hours. In experimental group, the number of colony was significantly different among 2, 6, and 24 hours, and between 2, 6 hours and 48, 72 hours (P<0.05), but no significant difference was found among 24, 48, and 72 hours (P>0.05). The biofilm thickness showed significant difference between the other time points (P<0.05) except between 48 and 72 hours (P>0.05). The percentage of viable cells in the outer layers of the biofilm was significantly higher than that in inner and middle layers at 48 hours (P<0.05). Three-dimensional reconstruction displayed that the surface of mixd species was uneven; living bacterium mainly located at the protuberance, and dead bacteria mainly located at the concaves. SEM image showed that Staphylococcus epidermidis attached to various forms of Candida albicans (spores, pseudohyphae, hyphae) gradually, and formed multilayer reticulate sophisticated structure on the surface of PVC with time. ConclusionCandida albicans-Staphylococcus epidermidis mixed species biofilm is sophisticated in structure. The combination of CLSM, SEM, and three-dimensional image reconstruction technology is ideal for investigation of mixed species biofilm on PVC material.
ObjectiveTo investigate the effect of the estradiol hormones on biofilm formati on and structure of Staphylococcus epidermidis after breast implant surgery. MethodsThe concentration of Staphylococcus epidermidis strains ATCC35984 was adjusted to 1×107 CFU/mL or 1×108 CFU/mL, and the type strains were incubated on the surface of silica gel in 125 pmol/L estradiol suspensions to prepare bacterial biofilms model in vitro. After cultured in vitro for 4, 6, 12, 24, 48, and 72 hours, bacteria growth and biofilm formation ability were assessed by means of the XTT and crystal violet staining respectively. According to the above results, the bacterial suspension concentration was selected for experiments. The experimental concentration of Staphylococcus epidermidis ATCC35984 suspension and the concentrations of 50, 125, 250, 500 pmol/L estradiol suspensions were mixed with silica gel respectively to prepare biofilm model in vitro, no estradiol suspension served as control group. The experimental concentration of Staphylococcus epidermidis ATCC12228 suspension was used to prepare the same model in the negative control. After cultured in vitro for 4, 6, 12, 24, 48, and 72 hours, the same methods were used to assess the bacteria growth dynamics and biofilm forming ability, and the scanning electron microscope (SEM) was used to observe bacterial biofilm structure cultured on the surface of silica gel; the laser scanning confocal microscope (CLSM) was used to measure bacterial biofilm thickness on the surface of silica gel after 6, 12, and 24 hours. ResultsAccording to the results of semi quantitative detection of crystal violet stain and XTT methods, the bacterial suspension of 1×107 CFU/mL was selected for the experiment. XTT results indicated that the growth rates of ATCC12228 strain (at 4, 6, 12, 24, and 72 hours) and ATCC35984 strain (at 4, 6, 24, and 72 hours) in 125, 250, and 500 pmol/L estradiol were significantly faster than those in 0 and 50 pmol/L (P < 0.05). The growth rate of 500 pmol/L group was significantly faster than 125 and 250 pmol/L groups at 4, 6, and 72 hours (P < 0.05), and the growth rate of 250 pmol/L group was significantly faster than that of 125 pmol/L group at 72 hours (P < 0.05), but there was no significant difference between 0 and 50 pmol/L groups (P>0.05). At the same time point and same estradiol concentration, the growth rates showed no significant difference between 2 strains (P>0.05). Semi quantitative detection of crystal violet staining showed no biofilm formed in ATCC12228 strain in all estradiol concentration groups at different time points. In ATCC35984 strain, the biofilm was found at 4 hours and gradually thickened with time, reached the peak at 24 hours. After cultured for 4 and 6 hours, the biofilm of 0 pmol/L groups were significantly thicker than that of 125, 250, and 500 pmol/L groups (P < 0.05). At 12 hours, the 125 pmol/L group had the thickest biofilm, showing significant difference when compared with other groups (P < 0.05). The CLSM showed ATCC35984 biofilm thickness of 125, 250, and 500 pmol/L was significantly less than that of 0 and 50 pmol/L groups at 6 hours (P < 0.05), but difference was not significant between other groups (P>0.05). Then the thickness of the biofilm increased gradually, and the thickness of 125 pmol/L group was significantly larger than that of other concentration groups at 12 and 24 hours (P < 0.05). The SEM observation showed that the biofilm of 125 pmol/L group was denser and thicker than that of the other concentration groups at each time point. ConclusionHigh level estradiol can promote bacteria growth, biofilm formation, and biofilm maturity of Staphylococcus epidermidis.
ObjectiveTo analyse epidermal growth factor receptor (EGFR) gene mutations in pathologically confirmed lung adenocarcinoma (LAC) samples obtained by endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA). MethodsClinical data of 964 consecutive patients who underwent EBUS-TBNA in Department of Thoracic Surgery, Fudan University Shanghai Cancer Center from April 2009 to September 2013 were retrospectively reviewed. EGFR gene mutations in 77 LAC patients who were comfirmed by cell morphology and immunohistochemistry were analyzed. There were 48 males and 29 females with their median age of 61 (range 33-78) years, and 43 patients were smokers. ResultsAll the 77 LAC patients were confirmed by immunohistochemistry. Among them, 31 patients (40.26%) were found to have EGFR gene mutations. There was no statistical difference in EGFR gene mutations between male and female patients (P=0.088). Mutation rate of EGFR genes of non-smokers was significantly higher than that of smokers (P=0.032). ConclusionSamples obtained by EBUS-TBNA can be used for EGFR gene mutations analysis. The mutation rate of EGFR genes of non-smokers is higher than that of smokers.
ObjectiveThis study aimed to explore the timing of the long-term antiepileptic drugs (AEDs) therapy in patients with stroke related seizures. MethodsWe enrolled 90 Patients with post-stroke seizures who diagnosed in neurology and epilepsy specialist clinic of Tianjin Medical University General Hospital and followed up for at least 12 months from September 2014 to August 2016. The patients were divided into early-onset seizure group (occurring within 2 weeks of stroke) and late-onset seizure group (occurring after 2 weeks of stroke).The two groups were subdivided into treated and untreated group after the first seizure. ResultsThe patients were followed up for 12~96m (median 20m). 31 patients in ES group, 19 of which in treated group and 12 of which in untreated group. 59 patients in LS group, 36 of which in treated group and 23 of cases in untreated group. The recurrence rate of second seizures occurred in each group and the comparison between the subgroups in the 3rd, 6th, 9th and 12th mouth of follow-up as follows. 1 LS group compared with the group of ES, the recurrence rate of second seizures was high (25.81%~38.71% vs. 49.15%~69.49%), and there was statistical difference (P < 0.05). 2 The recurrence rate of ES in untreated group was lower than that in untreated LS group (16.77% 33.33% vs. 56.52% 73.91%), but only in 3m and 12m the difference was statistically significant (P < 0.05). 3 There was on statistically significant different in ES treated group compared to untreated group, LS treated group compared to untreated group, ES treated group compared to LS treated group (P > 0.05). Both in group of ES and LS, The ratio of seizure recurred patients at different time points during follow-up period was highest at the time of 3m, 3 6m followed, within six months respectively as high as 91.67% and 76.59%. ConclusionOnly one early-onset seizure after stroke can be suspended long-term AEDs treatment, once it recurred that indicates the need for treatment. However, the recurrence rate of late-onset seizure was higher than that of early-onset seizure and it should be given long term AEDs treatment after the first seizure.