Objective To evaluate the feasibility to detect Prototheca in a mouse model of Prototheca zopfii cutaneous infection by using fluorescence in situ hybridization (FISH).Methods The model of Prototheca zopfii cutaneous infection was established by abdominal subcutaneous inoculation of Prototheca zopfii suspensions into 20 male BALB/c mice.Seven days after the inoculation,the mice were sacrificed,and tissue specimens were obtained from abdominal skin and subjected to microscopic examination,fungal culture and paraffin embedding.A PZ-probe was artificially synthesized and used to detect Prototheca in paraffin-embedded sections by using FISH.Moreover,both periodic acid-Schiff (PAS) and hematoxylin-eosin (HE) staining were performed to examine the paraffin-embedded sections.Skin specimens obtained from normal mice and Candida albicans-or Cryptococcus neoformans-infected mice served as the negative control.Results Clinical presentations,pathological examination and fungal culture results all confirmed the successful establishment of Prototheca zopfii skin infection model in mice.Prototheca was identified by FISH with the PZ-probe in the paraffin-embedded skin tissue sections from the murine model of Prototheca zopfii cutaneous infection,but not detected in the negative control tissue specimens,which was consistent with the results of PAS and HE staining.Conclusion FISH can be used to detect Prototheca in paraffin-embedded skin sections from the mouse model of Prototheca zopfii cutaneous infection.

Objective To develop a mouse model of cutaneous protothecosis.Methods Totally,48 BALB/c mice were randomly and equally divided into four groups:high-and low-concentration immunosuppressive groups-immunosuppressed mice inoculated with P.zopfii var.portoricensis suspension of 1 × 109 and 1 × 106 colony forming units (CFU) conidia/ml respectively,high-concentration healthy group-healthy mice inoculated with P.zopfii var.portoricensis suspension of 1 × 109 CFU conidia/ml,and control group-healthy mice inoculatedwith sodium chloride physiological solution.The P.zopfii suspension or sodium chloride physiological solution was subcutaneously inoculated to the abdominal skin of mice,with the inoculation volume being 200 μl.Skin appearance at the inoculation site was observed,and four mice were sacrificed in each group on day 7,14 and 28 after the inoculation.Skin specimens were resected from the inoculation sites of mice and subjected to pathological and mycological examinations.Results Mice in the three experiment groups inoculated with the P.zopfi suspension were all infected,with the appearance of papules and abscess at the inoculation sites of all mice as well as ulcer and crusts in some mice.Meanwhile,no mice were infected in the control group.Significant differences were noted in the diameter of skin lesions between the three time points in these experiment groups (all P ＜ 0.05),and the largest diameter of lesions was observed on day 7,which was (6.75 ± 1.09) mm in the high-concentration immunosuppressive group,(5.88 ± 1.17) mm in the low-concentration immunosuppressive group,and (5.96 ± 0.99) mm in the high-concentration healthy group.Comparisons of lesion diameter between the three experiment groups revealed a statistical difference on day 28 (F =8.91,P ＜ 0.05),with the largest lesion diameter observed in the high-concentration immunosuppressive group (4.38 ± 0.86 mm),but no statistical difference was found on day 7 or 14 (both P ＞ 0.05).Pathology of skin specimens consistently revealed necrosis,abscess and granuloma formation in the three experiment groups.Spores were found in the lesions of infected mice by haematoxylin-eosin staining,periodic acid-Schiff staining,and direct microscopy.Culture of tissue samples from the experiment groups grew Prototheca.Conclusion The mouse model of protothecosis can be established by subcutaneous inoculation of Prototheca suspension in the abdominal skin of healthy or immunosuppressed mice.

Objective To explore the long-term prognosis of the patientswithpulmonary arterial hypertension ( PAH) after cardiac repair operations through follow up .Methods We filtered the CHD patients with PAH after cardiac repair operation , admitted to the department of pediatric cardiology in Beijing Anzhen Hospital from June 2004 to December 2014, and collected their preoperative and postoperative data to analysis the long-term prognosis.Results 92 patients were included, median age was 20.78 years(2.44-60.26 years),median operation age was 13.54 years(1.33-53.99 years), World Health Organiza-tion functional classⅠ-Ⅱ was 80(86.96%), Ⅲ -Ⅳ was 12(13.04%), mean 6 min walk test distance was(481.74 ± 80.74) meters, mean Borg score was 1.53 ±1.14, and mean percutaneous oxygen saturation was 0.95 ±0.05.Among them, 64 patients completed preoperative right heart catheterization.Their average pulmonary artery pressure was(72.23 ±12.83) mmHg, aortic pressure on average was(85.57 ±11.91) mmHg, and average pulmonary vascular resistancewas(10.79 ±5. 04) Wood· U.The range of follow-up was between 6 months and 28.32 years(median 13.67years).During the follow-up, there were arrhythmia in 2 cases(2.17%), syncope in 7 cases(7.61%), hemoptysis in 4 cases(4.35%),heart failure in 8 cases(8.70%) and death in 1 case(1.09%).Compared with preoperative data, WHO cardiac function was found decreased in 22 patients(23.91%), stabled in 61 patients(66.30%) and significantly improved in 9 patients (9.78%).63 patients (68.48%) received targeted drug therapy, only 5 patients were treated with combined therapy,29 cases were not.Conclusion Compared with idiopathic pulmonary hypertension , PAH after defect correction has a lower incidence of arrhythmia , hemop-tysis and heart failure, and a higher long-term survival rate, and heart function is stable.To patients with PAH after defect cor-rection, targeted therapy is not only in perioperative period, but also more important in the long-term after surgery.In the targe-ted drug therapy, the combination treatment rate is low.

Background Epidemiologieal survey suggests that visual impairment in teenagers is a worldwide public health problem,and its leading cause is uncorrected refractive error.To conduct an extensive screening of visual impairment in teenagers and analyze the relevant affecting factors are significant works for the prevention and management of refractive errors.But high-quality epidemiological data on visual impairment and refractive error from large groups of children are lacking in Shanghai.Objective This study was to investigate the prevalence of ametropia and visual impairment in schoolchildren aged 6 to 12 years old.Methods In this cross-sectional study,4 686 students from 6 elementary schools in Baoshan District of Shanghai,a rural-urban fringe zone,China were selected by clustering sampling from May 2010 to April 2011.The eye examinations included visual acuity,ocular surface,ocular anterior segment,ocular media,fundus and intraocular pressure measurement,and the data autorefraction under cycloplegia and eye position were recorded.The cause of visual impairment was evaluated.Results A total of 4 594 students received examination with the response rate 98.0％,and autorefraction under cycloplegia was completed in 84.8％ schoolchildren (3 975/4 594).The prevalence rate of uncorrected visual acuity 0.5 or worse in both eyes was 14.4％ (662/4 594),with the wearing glasses rate 51.8％ (343/662).The percentage of students with the uncorrected visual acuity 0.5 or worse in at least one eye was 22.4％ (1 031/4 594).The prevalence rate of refractive error in the visual impaired students was 96.9％ (999/1 031),followed by amblyopia (37/1 031,3.6％).The prevalence rate of myopia,hyperopia,and astigmatic in the pupils after cycloplegia was 31.1％,4.3 ％ and 33.0％,respectively.In addition,the prevalence rate of myopia in public elementary schools was higher than that in migrant elementary schools (33.9％ versus 30.3％) (x2 =5.46,P =0.02).Logistic regression analysis showed that myopia was associated with age (OR =1.60,95 ％ CI:1.53-1.68,P ＜ 0.01) and female (OR =1.33,95％ CI:1.16-1.54,P＜0.01).Conclusions The leading cause of vision impairment is myopia in the elementary school students in Shanghai,China.The screening-ratio of refractive error and the coverage of refractive correction in the elementary school students in Shanghai are matters of urgent concern.

Objective To investigate the expression of miR-372 in the plasma of patients with acute my-eloid leukemia(AML)and the possible mechanism to participate in the development of AML. Methods Real-time quantitative PCR was used to detect the level of miR-372 in plasma. Bioinformatics software predicted the pos-sible target genes of miR-372 and dual luciferase reporter assay was performed to validate the prediction. In HL-60 cells,miR-372 was knocked down,and the effects on cell migration and cloning were detected by scratch test and clone formation. Results The level of miR-372 was significantly up-regulated in the plasma of AML patients. ROC analysis showed that miR-372 could distinguish between AML patients and healthy controls. Dual luciferase report-er assay showed that miR-372 could inhibit the activity of PTEN-3'UTR. Inhibition of miR-372 in HL-60 cells can significantly reduce the cell migration rate and clone formation ability. Conclusion In summary,for the first time,we showed novel data that the level of miR-372 was increased in the plasma of AML patients. By targeting the tumor suppressor gene PTEN,miR-372 may become a potential noninvasive biomarker for the screening and di-agnosis of AML.

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Objective: To analyze the prognosis and associated risk factors of pediatric idiopathic pulmonary arterial hypertension. Methods: A total of 119 patients under 18 years of age diagnosed as idiopathic pulmonary arterial hypertension in the Pulmonary Arterial Hypertension Center in Beijing Anzhen Hospital between June 2007 and May 2017 were enrolled in this retrospective study. The clinical informations and follow-up data were collected. The endpoints of follow-up were defined as death or undergoing lung transplantation. Kaplan-Meier survival curve was used to assess the survival,and the COX risk regression model was used to analyze the prognostic risk factors. Results: The mean age at diagnosis was (5.9±4.2) years. For 92 (77.5%) patients, the main reason for visit was decreased activity with shortness of breath after exercise. Seventy patients (58.8%) were in baseline NYHA functional class Ⅲ-Ⅳ and 49 patients (41.2%) were in NYHA functional class Ⅰ-Ⅱ. The mean systolic pulmonary arterial pressure estimated by echocardiography was (90±23) mmHg (1 mmHg=0.133 kPa) . Right heart catheterization was performed in 50 patients. Hemodynamic parameters revealed that the mean pulmonary artery pressure was (66±19) mmHg. Mean right atrium pressure was (8.5±3.4) mmHg. Mean pulmonary vascular resistance index was (17±9) wood·m² and the mean cardiac index was (3.4±1.3)L/m²; 100 patients (84.0%) received targeted therapy in which 55 patients (46.2%) were on monotherapy,40 patients (33.6%) were on dual therapy and 5 patients (4.2%) were on triple therapy. The mean time of follow-up was 22.0 months (0-108 months). During follow-up, 43 patients (36.1%) died and 1 patient received double-lung transplantation. Main causes of death including right heart failure, pulmonary hypertension crisis, asphyxia and massive hemoptysis. The mean survival time from diagnosis was 37.0 months,1-,2-,3-and 5-year survival rates were 86.3%, 72.2%, 51.4%and 37.8% respectively. Survival analysis showed that patients in baseline NYHA functional class Ⅰ-Ⅱ had better prognosis. COX regression analysis showed that NYHA function class, edema, increased total bilirubin and troponin concentration and the pulmonary artery and aorta diameter ratio measured by echocardiogram are risk factors of pediatric IPAH (HR=2.310, 2.723, 1.066, 1.696, 3.719, P=0.028, 0.005, 0.001, 0.024, 0.030) . While the existence of aterial septal defect or patent foramen ovale, using bosentan and phosphodiesterase inhibitors_, dual or triple therapy were protective factors (HR=0.563, 0.559, 0.603, 0.682, 0.044, P=0.169, 0.076, 0.115, 0.258, 0.220) . In multivariate analysis only edema associated with decreased survival (HR=2.398, P=0.025) . Conclusion Childhood idiopathic pulmonary arterial hypertension patients are seriously ill at visit. Worse cardiac function classification at visit associate with high mortality. Target therapy including using bosentan, dual or triple therapy can improve survival.

Objective To study the effect of vagus nerve stimulation (VNS) on the expression of multidrug resistance-associated protein (MRP1) in brains of rats with drug refractory epilepsy.Methods Rat models based on the lithium-pilocarpine epileptic rats were induced with sodium phenobarbital (PB).Then,the rats were randomly divided into three groups:VNS group (n=7),sham-VNS group (n=6) and control group (n=6); VNS through stimulation device was given to the rats in the VNS group,stimulation device without VNS was given to the rats in the sham-VNS group,and rats in the control group were only given conventional feeding.Behavioral changes and number of seizures were recorded by continuous video monitoring during the whole process.And the amount of MRP1 over-expression in each group was detected following a four-week VNS.Results (1) The seizure frequency in VNS group (2.6±1.0 per week) was significantly decreased as compared with that in sham-VNS group (5.3±1.1 per week) or control group (5.2±1.3 per week) after a four-week VNS (P＜0.05).(2) The MRP1 expression in VNS group had significantly statistical differences as compared with that in the sham-VNS group and control group (the absorbance values:9120±1496,19556±1462 and 20231±1710,P＜0.05).Conclusion VNS can effectively decrease the seizures frequency through reducing the MRP1 over-expression.

Objective To study the acute effect of vagus nerve stimulation (VNS) on status epilepticus (SE) in rats.Methods Forty-eight male Wistar rats were randomly divided into 4 groups:Group A (VNS preconditioning group),accepted VNS at 2 h before pilocarpine injection,Group B (VNS treatment group),accepted VNS immediately after pilocarpine injection,Group C (negative control group),implanted with electrode without stimulation,and Group D (SE model group),without electrode implantation (n=1 2).Lithium-pilocarpine was used to kindle the rats to establish SE models.Stimulation parameters used in the procedure of VNS were as follows:frequency,30 Hz; pulse width,0.5 ms; current,1 mA; and duty cycle,on-30 s,off-30 s.Behavior changes were observed and caspase-3 expression in dentate gyrus was detected 72 h after pilocarpine injection.Results (1) The seizure frequencies of grade Ⅳ and Ⅴ (Racine grading) in group A and B were significantly fewer than those in group C and D (P＜0.05); the seizure frequency of grade Ⅴ in group A was significantly fewer than that of group B (P＜0.05); SE latency in group A was significantly longer than that in other groups (P＜0.05).(2) Expressions of caspase-3 in the dentate gyrus of group A and B were significantly lower than those of group C and D (absorbancy values being 5854.7±856.5,6244.8±806.0,11957.0±1948.1 and 11543.2±1734.7,P＜0.05).Conclusion VNS has an acute seizure-suppressing effect on SE in rats and it can also reduce neuronal apoptosis induced by SE in hipocampus.

Objective To summarize the common causes and types of circuit failure after vagus nerve stimulator (VNS) implantation,and analyze the available methods of resolving these problems.Methods Three patients with postoperative circuit fault in the 97 patients performed VNS implantation for drug refractory epilepsy in our hospital from October 2011 to January 2014,and three patients with postoperative circuit fault performed surgery in other hospitals at the same period were chosen in this study.The causes of circuit failure of these 6 patients were identified by performing system diagnostics and X-ray evaluation.A revision surgery may be necessary if a broken or damaged lead was suspected.Results In these 6 patients of device failure,two patients were due to the disconnection of the lead from the pulse generator;the lead impedance became normal after re-inserting the existing lead connector pins into the pulse generator following proper lead insertion techniques.Four patients performed lead revision surgery to replace or remove the existing lead;in 3 of them,the helices and associated scar tissues were removed from the vagus nerve under the microscope,and a new set of electrodes was placed;in one of them,the lead was transected as much as possible and the distal portion of the lead was severed at the neck following the removal of pulse generator.There were no complications such as hoarse voice,dyspnea,slow rhythm,subcutaneous hematoma or infection after the revision surgery.Five patients were followed up,indicating the devices work normally.Four patients had obvious improvement,and one patient had no significant improvement after the revision surgery.Conclusions The high lead impedance is the main manifestation of circuit failure after vagus nerve stimulator implantation.Surgical exploration is an effective method to identify and resolve these problems.