Objective To study a multiple frequency tympanometry on ear system. Methods Based on the original low frequency and single ingredient SY-IA tympanometry, the study mainly focused on such aspects as multiple frequency probe tone circuit, constant pressure sound control system, micropressor control system and multiple frequency acoustic immitance & phase angle detection circuit. Results Multiple frequency tympanometry on ear system could detect multiple frequency acoustic immitance and phase angle for normal ear, tympanum loosed ear and ossicular chain break ear under different probe tone. Conclusion Experiments show that multiple frequency tympanometry on ear system based on ARM is helpful to the diagnosis of pathological changes which are insensitive to single low frequency tympanometry.

Objective To keep the intensity of sound constant and reduce the effect of ear canal difference on detection process.Methods Real-time detection of the probe tone in ear canal was carried out and by control ling the output circuit of the probe tone through the digital signal compensation algorithm,to realize real-time digital tympanometry of ear system and the function of constant pressure control.Results The results showed that the probe tone remained constant at 226 Hz for the measurement of middle ear and tympanometry was accurate.Conclusion The study simplified system configuration and provides basis for the clinical application.

Objective To reduce the error caused by the non-linearity of headphone and device via the design of calibration algorithm for acoustic immittance.Methods Values of calibration chambers with different volumes (0.5,2.0 and 5.0 ml) were measured using the digital detection system.Then,the calibration data tables,for linearity calibration with inputting the values of acoustic immittance,were produced by means of processing the signal of acoustic immittance using the digital calibration algorithm based on the values from the calibration chambers,Results With the low frequency probe tone measurement,it was confirmed that the range of error among various calibration chambers with different volumes was 1％.Conclusion The calibration algorithm enables the improving of the detection accuracy and provides the reliable data for further clinical application.

OBJECTIVEUsing the algorithm on the layered structure of the retina and quantitative analysis of the automatic segmentation technique is the key to the early diagnosis of glaucoma and other retinopathy on optical coherence tomography. Existing methods require high quality image and have low reliability. This paper used the improved complex nonlinear diffuse filtering and other methods to solve this problem.

METHODSThis paper includes algorithm such as automatic threshold, improved complex nonlinear diffusion filtering, morphological operations and peak detection. Use the method for the segmentation of 20 retinal layers images which acquired on the self-builder OCT system, the boundary of inner limiting membrane(ILM), outer nuclear layer(ONL), the photoreceptor segments(IS/ OS) and the RPE_ChCap layer are detected accurately. At last, the photoreceptor layer thickness is measured.

RESULTSThe results of segmentation and measurement are good corresponded with expert manual segmentation and measurements, retinal foveal measurements data is consistent with Zeiss Stratus OCT central retinal thickness 212+/-20 micro m.

CONCLUSIONThe algorithm proposed is prospective applied to clinical diagnosis of retinal diseases.

OBJECTIVEOptical coherence tomography images may be distorted by motion artifacts in dynamic imaging, so it may lead to misdiagnosis in clinical diagnosis. Motion artifacts correction has become an urgent issue in optical coherence tomography imaging.

METHODSFirstly, using the improved complex nonlinear diffusion preprocessing filtering reduced the noise of the sequence images, then using the image mass center aligned the distortion data in the Y direction, finally, using the method of deviation average corrected motion artifacts along the Y direction.

RESULTSAfter correction, the motion artifacts in the longitudinal 2D images and the 3D image disappeared, the surface of the 2D and the 3D image became more smooth, the structure between layers of the images got clear and distinct, retinal en-face single slice image was sharp, and the fundus tissue structure could be observed.

CONCLUSIONThe algorithm of correction makes the physical structure of the retinal display truly after motion artifacts correction.

Objective To observe the effects of high glucose and anoxia on Amot expression in vascular endothelial cells (VECs),and explore its role in angiogenesis.Methods VECs were incubated with different glucose concentrations for 48 h,and then cultured at normal oxygen concentration or anaerobic condition for 24 h.The protein expressions of p130-Amot and p80-Amot were detected by Western blot.After Amot expression was downregulated in VECs by siRNA,wound healing experiments and angiogenesis experiments were performed to test the effect of decreased Amot expression on angiogenesis.Results pl30-Amot protein expressions in low glucose (5.5mmol/L) plus normal oxygen group and low glucose plus anaerobic group were higher than those in high glucose (30mmol/L) plus normal oxygen group,high glucose plus anaerobic group,middle glucose (15 mmol/L) plus normal oxygen group,and middle glucose plus anaerobic group (all P＜0.01).Compared with low glucose plus anaerobic group,p130-Amot expression was higher in low glucose plus normal oxygen group (P ＜ 0.01).However,the expression of p80-Amot showed no statistically significant difference among different groups (P＞0.05).Compared to the normal VECs,the cells with decreased Amot expression by siRNA exhibited an attenuated cell migration in the wound healing experiments and a lesser tube formation in the angiogenesis experiments.Conclusions High glucose exerts a more significantly negtive effect on the Amot expression than anoxia in VECs.The downregulation of Amot expression inhibits migration and angiogenesis of VECs.

Objective To study the diagnosis and treatment of protracted bacterial bronchitis (PBB) in children. Methods Children with PBB conifrmed by bronchoscopy were recruited from May 2013 to April 2015 . The clinical data were retrospectively analyzed. Results All 31 cases include 18 boys and 13 girls were recruited. 28/31 were younger than 6 years old. They all complained of wet cough, some of them were reported with wheeze ( 17/31 ) and with ruttle in the lungs ( 16/31 ). White blood cell were in normal range ( 18/31 ) or slightly elevated ( 13/31 ). The C-reactin protein was in normal range ( 28/31 ). Chest X-ray test of 16 cases were normal. Twenty-four cases taken chest computerized tomograph scan, 5 had a sign of tracheobronchial stenosis. The purulent bronchitis without tracheobronchial stenosis were conifrmed by bronchoscopy. Four cases had tracheomalacia. The medians of proportion of neutrophil were 80% in bronchoalveolar lavage lfuid (BALF). The pathogens were identiifed in BALF in 17 cases, 6 with Streptococcus pneumoniae, 6 with Haemophilus parainfluenzae, 3 with Moraxella catarrhalis, 2 with Staphylococcus aureus and 1 with Haemophilus influenzae. The symptoms were improved in all cases and co-amoxiclav was prescribed to most cases when discharged. The course of antibiotics therapy was 2-4 weeks in 23 cases, and more than 4 weeks in 8 cases. Twenty-three ( 23 ) cases were cured but 8 of them relapsed. Another 8 cases were improved but not completely remitted, 7/8 were cured by further treatment for concomitant diseases such as nasosinusitis and allergic rhinitis. Conclusions Children with PBB are typically younger than six years old, and presented with prolonged wet cough and parent-reported wheeze, normal or with ruttle in the lungs. A conifrmed diagnosis was reached by bronchoscopy. The antibiotics therapy were effective, the course should be more than 2-4 weeks, however, relapse were common. When antibiotics therapy does not lead to complete remission, concomitant diseases should be considered.

Objective To investigate the clinical features and antibiotic susceptibility of osteomyelitis infected by Gram-negative bacteria (G-) in patients suffered from diabetic foot ulcers (DFU). Methods The clinical data of 91 DFU pa-tients accompanied with osteomyelitis (DFO) were retrospective studied. These patients hospitalized in the Tianjin Metabolic Diseases Hospital were divided into two groups, Gram-negative bacteria (G-) group (n=44) and Gram-positive bacteria (G+) group (n=42), respectively. The clinical features were compared between two groups. Logistic regression analysis was used to determine the risk factors for Gram-negative bactreial infection. The Gram-negative antibiogram was summarized. Results A total of 112 pathogens were isolated from 91 patients. G-bacteria were the most frequent pathogens (48.2%), following by G+ bacteria (47.3%) and fungi (4.5%). Pseudomonas aeruginosa was the majority of the G-bacteria. Comparing the two groups, the rate of antibiotic use within the previous 6 months was significantly higher in G-group (75.0%) than that of G+group (52.4%, P<0.05). There were no significant differences in the other indicators between two groups. The Logistic re-gression analysis revealed that the history of antibiotic use was the independent risk factor of G-bacterial infections in DFO patients. Antibiotics susceptibilities reflected G- bacteria were more prevalent to resist to cephalosporins and quinolonem, but sensitive to imipenem, ceftazidine and cefperazone-sulbactam. Conclusion Gram negative bacteria were not only the main pathogens isolated from DFO patients, but also frequently resistant to several popular antibiotics in China. The proper bacteria culture and antibiotic sensitivity test are especially emphasized to patients with DFU.

Objective To investigate SCCmec genotypes and drug-resistance profiles of the methieillin-resistant Staphylococcus epidermidis (MRSE) strains isolated from the patients suffered from diabetic foot infections (DFI) in the Tianjin Metabohc Diseases Hospital. Methods After dabridement, specimens of 390 infectious diabetic foot ulcers in the hospital from Jan 2008 to Jun 2010 were collected from the wound basal parts by cotton swab for culture. The disk-diffusion method was performed to examine antimicrobial susceptibility. DNAs of the MRSE strains were extracted, and their SCCmec genotypes were identified by PCR. Results Twenty of the seventy(28.6% ,20/70)Staphylococcus epidermidis strains were mecA posifive. Among the MRSE isolates, 2 ( 10.0% )were SCCmec Ⅱ ,9 (45.0%)were SCCmecⅢ and 9 (45.0%)were SCCmec Ⅳ. None of the isolates were genotyped as SCCmec Ⅰ or Ⅴ. No mater which genotypes they were, all the MRSE isolates were multi-drug resistant. They were resistant not only to β-lactams (including penicillins, cefoxitin and cephems), but also to non-β-lactams (including macrolides, fiuoroquinolones and sulfonamides ) . Resistance to voncomycin and rifampicin were not found in these strains . Conclusion SCCmec Ⅲ and SCCmecⅣ are major genotypes of the MRSE isolates from the infectious diabetic foot ulcers.The SCCmec Ⅳ genotype strains with multi-drug resistant profiles are prevalent in the diabetic foot infections.

Objective To prepare recombinant human epidermal growth factor (rhEGF) sustained-release microspheres and evaluate their morphology, rhEGF releasing activities and cell proliferation activity in vitro and compare difference of rhEGF sustained-release microspheres and rhEGF in facilitaring ulcer healing in diabetic rats. Methods (1) rhEGF sustained-release microspheres were prepared by the modified double emulsion method. Morphology of the microspheres was detected by transmission electron microscope and size distribution measured by laser granularity meter/Zeta electric potential meter. ELISA assays were applied to determine rhEGF releasing. (2)Proliferation of mouse fibroblasts was analyzed by MTr method. (3) Diabetic rat models were prepared and divided into four groups, ie, rhEGF sustained-release mierospheres group (Group A), rhEGF stock solution group (Group B), blank sustainedrelease mierospheres group (Group C) and PBS meustruum control group (Group D), which were given drug once a day. The wound healing rate was calculated by taking photographs at days 3,7,14 and 21. Skin specimens from the wound edge were harvested partially for observation of hydroxyproline (HYP) contents. Immunohistochemistry was employed to detect integrin 131 and keratin-19 and measure their positive staining area ratio. Results (1) The particle diameter of rhEGF sustained-release microspheres was 193.5 nm, with relative uniform particle diameter distribution. There showed no conglutination among rhEGF susrained-release microspheres, with good dispersibility. Releasing drug lasted for 24 hours and accorded with Higuchi release kinetic model. (2) Different concentrations of rhEGF sustained-release microspheres could promote the proliferation of mouse fibroblast, especially the concentration of 10 μg/L (P <0.05, compared with the control). (3) From the 7th day after treatment, Group A had the fastest wound healing rate, with statistical difference compared with other three groups (P < 0.05). Group A had higher HYP contents and positive area ratio of integrin β1 and keratin-19 than Group B. Conclusions rhEGF sustained-release microspheres prepared by the modified double emulsion method have uniform particle size and can last release for 24 hours. Compared with rhEGF stock solution, rhEGF sustained-release microspheres have faster and better ulcer healing and higher healing quality in diabetic rats.