Objective: To optimize the sample pretreatment and establish an ultra-high-performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap-MS) assay based on the parallel reaction monitoring (PRM) mode for determination of antibiotic residues in chicken meat. Methods: Blank matrix-spiked chicken meat samples were extracted with 95% acetonitrile aqueous solution containing Na2EDTA and formic acid. The extraction solutions were cleaned up using different combinations of C18, PSA and GCB fillers, and the combinations with a higher antibiotic recovery rate was screened. The residues of 32 antibiotics were determined using UPLC-Q-Orbitrap-MS based on the PRM mode. Results: If the extraction solution was cleaned up using the C18 filler, the largest number of antibiotics with a spiked recovery rate of >80% was seen, with matrix effects of 82.2% to 112.6%. The detection limits of 32 antibiotics were 0.8 to 5.8 μg/kg, with linear correlation coefficients of >0.99, spiked recovery rates of 71.3% to 111.5% and relative standard deviations of 3.2% to 14.2%. Conclusion The UPLC-Q-Orbitrap-MS assay is suitable for determination and quantitative analysis of multiple antibiotics in chicken meat. Key words： high-resolution mass spectrometry orbitrap antibiotic residue parallel reaction monitoring

The repair of articular cartilage defects has been a dif ficult problem. Presently cartilaginous autograft and allograft, periosteal and perichondral graft and transplantation of chondrocytes had been widely applied t o repair cartilaginous defects. The application of tissue engineering might be a method of hope. This article reviewed the progression of cartilage transplantation and the tissue engineering.

Objective To probe imaging manifestations of bronchogenic cysts,as that to improve the knowledge of this disease.Methods Imaging features of congenital bronchial cyst in 32 cases confirmed by operation and pathology were analyzed.Results 4 cases of cyst localized in mediastinum ,all were liquid cyst.28 cases of cyst localized in lungs,aqmong them,22 cases were air-liquid cyst and 6 cases were air cyst,the diameter of lesions was 1~10 cm,there were 2 cases with pyothorax,4 cases with pulmonary maldevelopment,one case with brochio-pleural fistula.The cysts appeared as round or similar round,the walls of cyst was thin.The wall of cyst was obscure and there was infiltrating shadow around cysts when in company with infection.Conclusion X-ray and CT are very important in diagnosing congenital bronchial cysts,CT scan is of very help in diagnosing the compley cysts.

Objective To analyze the clinical features,and prognosis of the interstitial lung disease (ILD) in patients with dermatomyositis (DM) and polymyositis (PM) by chest X-ray,chest high-resolution CT scan (HRCT) and pulmonary lung function.Methods Thirty-three patients hospitalized with DM/PM associated ILD were retrospectively analyzed.Results Thirty-three patients with ILD were confirmed by HRCT.Abnormal pulmonary function tests were available in 82% of patients.Clinical-imaging analysis revealed that the pathological features of ILD were non-specific interstitial pneumonia (NSIP,57%) and unusual interstitial pneumonia (UIP,25%).UIP types showed a poor prognosis and high mortality (70%).Conclusion This study shows that HRCT is more sensitive for the diagnosis of ILD than lung function tests and chest X-ray.Combined HRCT and chest X-ray with lung function tests and blood gas analysis have shown that the major pathological types of ILD are NSIP and UIP,in which UIP are associated with high mortality and poor prognosis.

Objective To evaluate the therapeutic effect of etanercept in patients with Behcet's disease (BD). Methods Twenty six patients with active BD were eligible for anti-TNF-α treatment, which failed previously to commonly used drugs. All the patients were stopped previous therapy and treated with etanercept 25 mg twice a week. The clinical responses and laboratory parameters were evaluated at the baseline, week 2, 4, 8, 12 and 24, and week 12 of etanercept withdrawal. Results Etanercept was effective in several clinical lesions and lab. parameters at all time points. Especially at week 2, including oral ulcers, genital ulcers, erythema nodosum, pseudofolliculitis and swollen joints (P<0.01). Two of four cases of uveitis were contro-lled with addition of topical corticosteroids; The ileocecal junction ulcer of 1 patient healed after 2 months etanercept therapy; four cases of epididymitis reached complete remission at week 4. ESR(P=0.012) and CRP (P=0.013) were also rapidly decreased at week 2. Most patients were relapsed at week 12 of etanercept withdrawal compared with baseline except for swollen joints (0.65±1.09 vs 0.31±0.60, P<0.01) after etanercept withdrawal, in which the typical rebound was CRP from (481±312) mg/L to (549±267) mg/L (P=0.013).Conclusion Etanereept has rapid efficacy onset for most of the mucocutaneous disorders and possibly effective for arthritis, ophthalmitis, epididymitis and colonic ulcer of BD.

Objective: To determine human tear proteins using nanoliter liquid chromatography coupled to quadrupole orbitrap mass spectrometry (NanoLC-Q-Orbitrap-MS), and perform a bioinformatics analysis of main proteins. Methods: Human tear samples were collected with capillary, transferred to 3 kDa ultrafiltration tubes containing 400 μL of superpure water and centrifuged at 12 000×g for 15 min. Repeated extraction of tear proteins were performed four times, and following digestion with trypsin, the proteins were separated using the Waters NanoAcquity peptide BEH C18 column (1.7 μm, 100 μm×100 mm) and determined using NanoLC-Q-Orbitrap-MS with the mobile phase of 0.1% formic acid aqueous solution and acetonitrile (0.1% formic acid) in the full MS/dd-MS2 mode. The types of proteins were characterized in the Uniprot database using the software Proteome Discoverer version 2.1 and verified using bovine serum albumin. The tear proteins were subjected to gene annotation analysis using the String database. Results: A total of (387±160) human tear proteins were yielded, with a relative standard deviation (RSD) of 4.13%, and there were 25 types of proteins with a relative high abundance, including lipocalin 1, lysozyme and lactoferrin. The peptide sequence coverage of bovine serum albumin was (86.08±2.61)%, with a RSD of 3.03%. The 25 major tear proteins were involved in substance transduction among cells, homeostasis process, negative regulation of the endopeptidase activity, detection of chemical stimulants and humoral immune responses, and the 16 proteins had close interactions. Lacritin, lipocalin 1, lactoferrin, lysozyme and zinc-α 2-glycoprotein, which had a relative high abundance, had close biological connections. Conclusion NanoLC-Q-Orbitrap-MS is stable, reliable and feasible for detection of multiple proteins in tears.

Objective To evaluate the treatment results and psychological changes in female patients with stress urinary incontinence (SUI) before and after TVT-O.Methods Fifty-six female patients with SUI underwent TVT-O from Nov.2011 to Mar.2012.Preoperative and postoperative International Consultation on Incontinence Questionnaire Short Form (ICIQ-SF) score,Incontinence Quality of Life Questionnaire (I-QOL) score,one hour urine pad test,Self-rating Anxiety Scale (SAS) score,and Self-rating Depression Scale (SDS) score were recorded and analyzed.Results Followed up for 3 months,the subjective satisfactory rate was 94.6％ (53/56),the objective efficiency rate was 91.1％ (51/56),and there was no recurrence in this group.Before operation and 3-month after operation,ICIQ-SF scores were 15.9±3.2(13-20) and 8.6±2.7 (4-13),t=6.02,P=0.01 ; I-QOL scores were 40.3±18.9 (33-62) and 87.6±11.2(76-102),t=6.81,P=0.01; The urine leakages in one hour urine pad test were 26.7±7.1 (9-51)g and 5.2±2.5 (1-8)g,t=7.13,P=0.00; SAS scores were 59.3±8.3 (48-72) and 42.6±4.6 (39-52),t=9.15,P =0.01; SDS scores were 57.2±5.6 (49-69) and 40.2±3.4 (36-54),t=8.63,P=0.01,respectively.No serious and long-term complication occurred.Conclusions TVT-O is a safe and effective treatment option for female SUI.This treatment can improve the quality of life and psychological status of the patients.

Objective To study the correlation between the risk factors of coronary slow flow phenomenon ( CSF) and the level of plasma D-dimer in patients with ACS ( acute coronary syndrome) after emergency percutaneous coronary intervention ( PCI) .Methods A total of 297 patients with ACS after PCI were enrolled for retrospective analysis.All patients were divided into CSF group and control group in the light of corrected thrombolysis in myocardial infarction (TIMI) frame count method (cTFC).Multivariate analysis for evaluating clinical predictors of CSF was carried out using Logistic regression test and Pearson analysis to find the correlation between plasma D-dimer and cTFC.The predictive value of D-dimer level in the occurrence of coronary slow flow was determined by using receiver operating characteristic ( POC) curve analysis.Results CSF was observed in 59 cases (19.8%).The plasma D-dimer was significantly higher in the coronary slow flow group compared with the control group ( P <0.05 ) .Multivariate regression analysis and Logistic regression test showed that the level of plasma D-dimer ( OR =1.276, 95%CI:1.132-3.224, P=0.012), thrombus score (OR =1.108, 95%CI: 1.085-2.103, P =0.018) and target lesion length of culprit vessel ( OR =1.436, 95%CI: 0.635-1.382, P =0.037 ) were the risk factors of CSF.Correlation analysis showed that plasma D-dimer were positively associated with CSF. Receiver operating characteristic ( ROC ) curve analysis showed that D-dimer cutoff point at 515.3 ng/ml had a good judgment significance ( AUC 0.783, OR =1.502, 95%CI: 1.324-2.531, P =0.005). Conclusions The increased D-dimer level is a risk factor and plays an important role in the ACS patients with the CSF phenomenon, thereby predicting no-reflow phenomenon after primary PCI in these patients.

Objective : To develop an analytical method of ibotenic acid (IBA) and muscimol (MUS) in wild mushroom by dansyl chloride (DNSCl) derivatization-liquid chromatography-tandem mass spectrometry (LC-MS/MS), and to provide technical support for etiological identification of mushroom poisoning events. Methods : The sample was extracted with hydrochloric acid solution, derived by bimolecular DNSCl, diluted and inorganic salts precipitated with acetonitrile. The extract was separated by a waters XBridgeTM BEH C18 column and measured by LC-MS/MS. Results : The limits of detection for IBA and MUS in wild mushroom were 0.15 mg/kg and 0.1 mg/kg, respectively. Good linear relationship was obtained for IBA and MUS at the range of 0.5-250 mg/kg with the correlation coefficient of 0.997 and 0.999, respectively. The average recoveries at three spiking levels were 84.5%-102.0% with relative standard deviations (RSDs, n=6) of 4.7%-8.6% for IBA. The average recoveries were 88.6%-95.4% with RSDs (n=6) of 4.9%-7.5% for MUS. Conclusion The optimized sample extraction and bimolecular DNSCl derivatization conditions can achieve rapid and accurate analysis of IBA and MUS in wild mushroom poisoning sample.

Objective: To optimize the enzymatic digestion conditions of trypsin, so as to improve the testing capacity of mass spectrometry for shrimp allergens. Methods: The enzymatic digestion test was carried out by response surface methodology for optimizing pH, temperature and time. After enzymatic hydrolysis, the peptides were separated by chromatography and determined by high-resolution mass spectrometry with Q-orbitrap. The allergen protein was identified and quantified by UniProt database and MaxQuant software. Results: Two allergen proteins, tropomyosin and arginine kinase, were isolated from shrimp, and their intensities ranged from 100.2×106 to 436.5×106. Response surface analysis showed that when the digestion time was 4.29 hours, the temperature was 44.15 ℃ and pH value was 6.55, the maximal intensity of the allergen proteins was 457.48×106. The experiment was validated with the digestion time of 4.2 h, pH value of 6.5, and temperature of 44 ℃, then resulted in the average intensity of 448.1×106. The deviation from the predicted value was 2.1%. Conclusions The conditions of enzymatic digestion can be optimized by response surface methodology. The enzyme may have the best performance with the pH value of 6.5, temperature of 44 ℃ and digestion time of 4.2 hours.