BackgroundPosterior capsule opacification(PCO) is the main cause inducing low vision after extacapsular cataract extraction. Our previous study determined that polylysine-ethylene diamine tetraacetic acid (EDTA) (PLE) can suppress the incidence of PCO. ObjectiveThe goal of this experiment was to investigate the inhibition of polylysine-EDTA on rabbit lens epithelial cells (LECs)proliferation in vitro and the effective concentrations of polylysine-EDTA. MethodsThe anterior capsular membranes from 10 3-month-old clean New Zealand white rabbits were digested and then cultured to obtain the LECs. The second and third generation of LECs were inoculated on the 96-hole culture plate with the cell density of the 1 × 105/ml. 12.5,25.0,50. 0,100. 0 μmol/Lof PLE were added into the culture medium for 48 hours respectively,and the DMSO medium was used at the same way as the control group. The proliferation of the LECs was then detected by MTT method and the inhibitory rate of PLE on LECs growth was calculated. ResultsLECs grew at a near normal state in ≤25.0 μmol/L PLE groups,however,cultured LECs were out of shape and the numbers decreased with the weakened adhesion ability in ≥50.0 μ mol/L PLE groups. The A490 values of LECs were 0. 278±0. 013,0. 266±0. 028,0. 260±0. 022 and 0. 247±0. 012 in 12. 5,25.0,50. 0, 100. 0 μmol/L polylysine-EDTA groups respectively and were lower than 0. 311 ±0. 038 of DMSO control group( P=0. 035,0. 011,0. 009,0.013 ). The inhibitory rates of 12. 5,25.0,50. 0, 100.0 μmoL/L PLE on LECs proliferation were 10.61％ , 14.47％ , 16.40％ and 20. 58％ respectively. ConclusionsPolylysine-EDTA can inhibit the growth and proliferation of LECs in vitro at a dose-dependent manner.

Objective To perform a DNA-based prenatal diagnosis in a family with recessive dys-trophic epidermolysis bullosa, and to develop a strategy to eliminate matemal cell contamination in arnniotic fluid samples. Methods Amniocentesis was carried out at gestation week 16, amniotic fluid culture was used to separate fetal cells from maternal blood cells. Peripheral blood was obtained from the proband, and her parents. Genomic DNA was extracted from peripheral blood and aminotic cells. Subsequently, PCR and direct sequencing were performed to detect pathogenic mutations in the COL7A1 gone. Karyotype analysis was used to confirm paternal information in amniotic fluid. Linkage analysis between micro-satellite markers was performed to confirm the fetal genotype. Resulta Centrifugation showed visible contamination of aminotic cells by blood cells. Direct sequencing revealed that the proband was a carrier of both maternal mutation, R525X in exon 12, and paternal mutation, R2610X in exon 105, while the fetus only carried the maternal mutation, R525X. The second direct sequencing and hapiotype analysis after elimination of mater-nal blood cells by amniotic fluid culture confirmed that the fetus was a carrier of maternal mutation with nor-real phenotype. The pregnancy continued and a clinically unaffected girl was born at gestation week 40.Conclusion The accuracy of DNA-based prenatal diagnosis could be improved by the combination of direct sequencing, amniotic fluid culture, karyotype analysis and linkage analysis, etc.

Objective To evaluate the clinical effect of the intertrochanteric fractures with or without lateral femoral wall fractures using proximal femoral nail antirotation (PFNA).Methods From May 2008 to June 2011,102 patients with intertrochanteric fractures were treated with PFNA.In accordance with the preoperative three dimensional CT reconstruction(3D CT) images,the group A included 41 cases with lateral femoral wall fractures,and the other 61 cases with an intact lateral wall were in group B.According to the AO/OTA classification,there were 5 cases in 31-A2,36 in 31-A3 in group A,and 61 in 31-A2 in group B.The operative time,operative blood loss,average length of stay,postoperative X-ray images,and 3D CT images were collected for each patient.Time of partial weight-bearing,full weight-bearing and fracture healing were also recorded.Clinical evaluation was made using the functional recovery scale (FRS) of hip fractures.Results The mean operation time was 56±8 min in group A vs 45±6 min in group B; the mean blood loss was 238±21 ml vs 175±11 ml; the average length of stay was 17±3 days vs 15±3 days.On the postoperative radiography,the blowout of lateral trochanteric wall only occurred in 8 (19.5％,8/41) cases in group A and 3 (4.9％,3/61) in group B.According to the postoperative 3D CT,the similar findings were seen in 36 (87.8％.36/41) cases in group A and 45 (73.8％,45/61) in group B.Eighty-two cases were followed up for 6 to 35 months (mean.19.5 months).The mean FRS score was 64.2±4.8 points in group A and 76.5±7.9 points in group B.Conclusion When treating unstable intertrochanteric fractures,iatrogenic fractures in lateral trochanteric wall could be easily caused with using PFNA.3D CT could effectively evaluate iatrogenic trauma in the intertrochanteric fractures.

The new intein-mediated PHB purify protein system is a high expression, automatic cutting, for purification, low-cost protein purification system,it is conducive to large-scale protein purification.Choose human antibacterial peptide LL-37 as the purification objects,which is poison to prokaryotic cell.We construct intein-mediated PHB purified human antimicrobial peptide LL-37 system through genetic engineering technology and use this system to purify LL-37. The results show that this system can highly express LL-37 fusion protein and purifiy the product as same size with expectations.

Objective To investigate the long-term outcome of children with recurrent wheeze and to determine the effectiveness of inhaled hormone therapy. Methods One thousand and thirty-five children with recurrent wheezing were followed up for more than 4 years and the data were retrospectively evaluated. Results Of 1035 cases, 751 (72.56%) patients outgrew their wheeze during the follow-up period, whereas the other 284 (27.44%) patients had recurrence wheeze during the last two years. The age of wheezing onset was<3 years in 542 (52.37%) cases, from 3 to 7 years in 386 (37.29%) cases, and from 7 to 12 years in 107 (10.34%) cases. There was significant difference in clinical control rate among groups with different wheezing ages onset (χ2=45.27, P<0.001). Children with wheezing age onset from 7 to 12 years had the lowest clinical control rate. Among 1035 wheeze children, 343 (79.95%) children in 429 cases who received inhaled hormone therapy for more than one year outgrew their wheeze. Whereas 408 (67.35%) in 606 cases who did not receive inhaled hormone therapy outgrew their wheeze. There was significant difference of clinical control rate between inhaled group and non-inhaled group (P<0.01). Con-clusions The age of wheezing onset is<7 years in 89.66%of children with recurrent wheeze. Most of them can be clinicalycon-trolled. The long term inhaled hoemone therapy for children with recurrent wheeze can reduce the risk of developing adulthood asthma.

Objective Digital PCR ( dPCR ) was established to detect plasma epidermal growth factor receptor (EGFR) T790M mutation of non-small cell lung cancer (NSCLC) patients and was evaluated in terms of analytical performance and clinical application significance.Methods The specific primers and probes for EGFR T790M mutation and wildtype were designed to establish dPCR platform.Limit of blank, sensitivity and linearity of dPCR were evaluated by the detection of plasmids with different concentrations to set up optimal reporting system and reanalyzing process.The mutation of EGFR T790M in plasma and tissue samples from 10 patients with advanced NSCLC resistant to EGFR-TKI therapy who were enrolled in Zhongshan Hospital Fudan University from January 2014 to October 2015 were analyzed by dPCR and amplification refractory ( ARMS) , respectively.The consistency was evaluated between dPCR and ARMS by Chi-square test.The correlation of T790M abundance detected by dPCR between plasma and tissue samples was also analyzed by Peasrson correlation analysis.Results Limit of blank and sensitivity of dPCR was 10 copies and 0.01%, respectively.dPCR was evaluated as linear in the range of 0.01%-100%( Y=1.226X-3.984,R2 =0.999 ).The consistency between dPCR and ARMS of tissue samples was good ( kappa=0.80), while the positive rates of plasma T790M detected by dPCR was significantly higher than ARMS (50%vs 20%,P<0.05).It was found that T790M abundance detected by dPCR was highly correlated between lung cancer tissue and plasma ( R =0.923, P <0.05 ) using Pearson correlation analysis. Conclusions A new method of dPCR with high sensitivity and absolute quantification is established for the detection of EGFR T790M mutation in plasma from advanced NSCLC patients, which brings tumor liquid biopsy into real.It has the ability to provide the most direct and valuable guidance for clinicians to make decision on EGFR tyrosine kinase inhibitors therapy in patients with advanced NSCLC resistant to EGFR-TKI.

Objective To understand the pathogenesis of primary hypothyroidism associated with polymyositis-like syndrome.Method Four cases were presented and the clinical characteristics were analyzed.Results All of the four patients were male.Complaints of weakness primarily affecting proximal muscles and elevation of muscle enzymes were presented.Primary hypothyroidism was diagnosed after clinical examination.Complaints of weakness and the elevated muscle enzymes were all improved after the substitution of thyroxine.Conclusion Hypothyroidism could cause polymyositis-like syndrome,and thyroid function should be screened when elevation of muscle enzymes is presented.

Objective To analyze the results of survival and local regional control of early-stage breast cancer patients treated with breast-conserving surgery and whole breast radiation therapy, and to explore the factors influencing local control. Methods From October 1995 to September 2005,335 patients with early-stage breast cancer were treated with breast conservative therapy, including 25 with post-operative radiotherapy alone( group A), 194 with sequential chemo-radiotherapy( group B) ,80 with concurrent chemoradiotherapy( group C), and 36 with chemo-radio-chemotherapy (group D). Axillary dissection was performed in 312 patients and sentinel node biopsy in 2. Adjuvant chemotherapy was given to 310 patients. Radiation of 50 Gy in 25 fractions was delivered to the whole breast in 256 patients,and to the breast and regional nodes in 79. Another 10 Gy in 5 fractions was boosted to the tumor bed. The proportion of patients with surgery-radiation interval of less than 4 weeks,4-8 weeks,8-20 weeks and over 20 weeks was 16.4%, 23.6% ,37.3% and 22.7% ,respectively. Concurrent chemo-radiation and" sandwich" modality were delivered in 80 and 36 patients,respectively. Results The median follow-up was 48(25.3-146.7) months. The 5-year local-regional control and ipsilateral breast control rates were 94.5% and 95.6%, respectively. The 5-year disease-free survival,metastasis-free survival and overall survival rates were 88.6% ,93.2% and 98.8% ,respectively. Lymph-vascular invasion and neural invasion were significant factors influencing local control in univariate analysis. No significant differences were found in local control rates among different surgery-radiation intervals or different sequencing of radiotherapy and chemotherapy. Multivariate analysis showed that lymph-vascular invasion and reexcision for positive or unknown margin were independent prognostic factors influencing local control. In group A + B, C and D, the frequency of grade Ⅲ skin toxicity was4.5%.80.0%.and 77.0%,respectively(χ2=226.00,P=0.000).Conclusiom Breast conservative therapy for early-stage breast eancer results in good local-regional control and overall survival.Lymph-vascular invasion and reexcision are independent prognostic factors for local control.Surgery-radiation interval and sequencing of radiotherapy and chemotherapy have no impact on local contr01.Grade Ⅲ skin toxicity of the concurTent radio.chemotherapy and the chemo.radio.chemotherapy group is significantly higher than that of the sequential chemotherapy-radiotherapy plus radiotherapy alone group.

Objective To evaluate the effect of open reduction and internal fixation in treatment of calcaneal fractures in aged patients. Methods Between January 1997 and June 2007,37 patients (41 fractures)at a mean age of68.3 years(60-78 years)with calcaneal fractures underwent surgical operations.According to Sanders classification based on CT scanning.14 fractures were rated as typeⅡ,18 as type Ⅲ and 9 as type Ⅳ.Of all,5 fractures were treated with reconstruction plate,18 with shapeable titanic plate,4 with Y-shaped plate,12 Depuy titanic plate and 2 with AO locking titanic plate plus open reduction and internal fixation.The foot function was evaluated by X-ray,American Orthopedic Foot and Ankle Society(AOFAS)Score and Maryland Foot Score. Results A total of 27 patients (60.9%)with 31 fractures were followed up for 12-48 months(average 27.6 months),which showed that all fractures were healed,with morphous improvement of the calcanus determined by 8 parameters measured in X-ray films.The average active range of motion was 36°of plantar flexion,12°of dorsiflexion,15°of inversion,and 10°of eversion.According to Maryland Foot Score,functional foot score was excellent(90-100 points)in 16 fractures,good(75-89 points)in 13 and fair(50-74 points)in 2,with excellence rate of 94%(29/31). Conclusion Open reduction and intemal fixation can gain good clinical result in severe calcaneal fractures in aged patients with obvious dislocation of posterior articular facet.

Objective To explore the clinical characteristics of cardiac involvement in children with mitochondriopathies.Methods The clinical data of 23 children with mitochondriopathies were reviewed.The changes of electrocardiography,echocardiography and heart enzymes were analyzed.Results In 15 cases of mitochondrial encephalomyopathy,lactic acidosis,and stroke-like episode(MELAS syndrome),electrocardiography was performed on 9 cases,6 of them showed abnormal electrocardiographic findings,including right bundle branch block,ST-T change,Wolff-Parkinson-White syndrome,et al.On echocardiographic examination in 9 MELAS syndrome ca-ses,only 1 case showed hypertrophy cardiomyopathy.Six cases had increased plasma creatine kinaseMB(CK-MB) mass and only one of 12 MELAS syndrome cases had increased cardiac troponin I(cTnI) level.In 8 cases of subacute necrotizing encephalomyopathy(Leigh syndrome),electrocardiography was performed on 5 cases,4 of them showed abnormal electrocardiographic findings,including sinus tachycardia,ST-T change and low voltage.Two cases showed normal electrocardiography.Three out of 6 cases with Leigh syndrome showed increased plasma CK-MB mass.The molecular genetic examinations were performed in 13 cases of MELAS syndrome and 6 cases of Leigh syndrome.The mitochondrial DNA nt 3243 A→G mutation was found in white blood cells of 9 MELAS syndrome cases,the mutation rate being 37%-60%.The mitochondrial DNA nt 8993 T→C mutation was found in white blood cells of 2 Leigh syndrome cases.Conclusion In children with mitochondriopathies,myocardiac involvement is comparatively common,and even cardiomyopathy can occur.