Objective:To analyze the characteristics of long non-coding RNA (lncRNA) expression in nonalcoholic fatty liver disease (NAFLD).Methods:lncRNA-mRNA microarray was conducted on the liver tissue samples from 10 patients with simple gallbladder stone (5 NAFLD liver samples and 5 normal liver samples),and the differentially expressed lncRNA was analyzed by bioinformatics technology.Results:Compared with the normal liver samples,there were abnormal expression of 1 735 lncRNAs and 1 485 mRNAs in NAFLD liver samples.Among them,535 lncRNAs and 760 mRNAs were up-regulated,1 200 lncRNAs and 725 mRNAs were down-regulated.Conclusion:Compared with normal liver,the expression oflncRNA in NAFLD tissues is obviously abnormal.These lncRNAs may play an important role in the occurrence and development of NAFLD.

Objective To study the effect of lateral supramalleolar perforator flap for skin and soft tissue defect of lateral malleolus.Methods From January 2012 to December 2015,36 patients with soft tissue burn of ankle in the Traditional Chinese Medicine Hospital of Cixi were selected as study objects.The average wound area was 4 ～ 10cm.The average flap size was 5 ～ 13cm.There were 17 cases received skin flap of behind perforator of the lateral malleolus.There were 19 cases received skin flap of front perforator of the lateral malleolus.The effect of repair was observed.Results All flaps survived in 36 cases after operation.There were 7 cases of 2.6cm of blood stasis,and necrosis occurred in 2 cases which healed after dressing change.Transplantation of skin flap and the original skin had good adhesion,skin color and texture was similar to the surrounding skin during the follow-up period of 6 to 12 months.The survival time and walk time of the repair of anterior perforator flap were (2.20 ± 0.70) weeks,(4.70 ± 1.10) weeks,respectively,which were longer than those of the lateral retromalleolar perforator flap [(2.10 ± 0.80) weeks,(4.20 ± 1.10) weeks],but the differences were not statistically significant (all P ＞ 0.05).The American orthopaedic foot and ankle society(AOFAS) score of the anterior perforator flap was significantly lower than that of the lateral retromalleolar perforator flap,the difference was statistically significant[(77.47 ± 3.41) points vs.(80.12 ± 4.12) points,t =2.110,P ＜ 0.05].Conclusion Lateral supramalleolar perforator flap has significant effect for skin and soft tissue defect of lateral malleolus.The operation is simple,the success rate is high,it is worth to be popularized in clinical.

Objective To establish and optimize a method for simultaneous determination for the contents of sodium,potassium,calci-um and magnesium ions in serum using microwave digestion combined with ion chromatography,and evaluate its performance.Methods Serum samples were pretreated by microwave digestion,and four cations were quantitatively determined using ion chromatography.The linearity,precision,recovery and accuracy of the established method were verified with reference to the EP6-A and EP15-A3 docu-ments issued by the Clinical and Laboratory Standards Institute(CLSI).Results The imprecisions of determinations for sodium,po-tassium,calcium and magnesium ions expressed as coefficient of variation(CV)were 0.10％to 0.36％,0.15％to 0.48％,0.22％to 0.87％and 0.21％to 0.73％,respectively.The recoveries with adding standard of the four cations were between 97.3％and 103.0％.Referred to RELA2020 and RELA2021,the deviations of the analysis results were-0.57％to-0.14％,-0.84％to-0.16％,-1.82％to-0.37％,and-0.60％to 0.34％for sodium,potassium,calcium,and magnesium,respectively,and all the comparisons were pas-sed.The limits of detection/quantification were 0.017 5/0.058 3 mmol/L,0.000 5/0.001 7 mmol/L,0.009 9/0.032 9 mmol/L and 0.001 1/0.003 5 mmol/L for the four cations,indicating that after pretreatment,no significantly interfere from other impurities for the results of determination was found.Conclusion A method for simultaneous determination of sodium,potassium,calcium and magnesi-um ions in serum was successfully established and optimized using microwave digestion combined with ion chromatography.The method showed good precision and accuracy,simple operation,short time consumption and low cost,thus it could be used for quality improve-ment of clinical electrolyte testing in serum.

Objective:To develop a candidate reference method for cortisol in human serum by isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC/MS/MS).Methods:An isotope-labeled internal standard was added to samples, followed by alkalizing with sodium carbonate solution and liquid-liquid extraction with n-hexane/ethyl acetate. Isoelution with a methanol aqueous solution was employed for liquid chromatographic separation, while ESI in the positive ion and multiple reaction monitoring mode were used for mass spectrometry. The volume of sample, standard, and internal standard solutions were all controlled by weighing, and the results were calculated by bracketing method. The accuracy, precision, specificity, linearity, LOD and LOQ of this method were evaluated referring to the CLSI C62-A and C50-A guidelines and the domestic expert consensus documents.Results:The method demonstrated excellent accuracy and precision with the bias of ERM-DA192, ERM-DA193 and RELA 2021-2023 all being less than 2%. The recovery of added cortisol ranged from 98.2% to 101.1%. Both intra- and inter-assay imprecisions was <2%. The method was free from interference by structural analogue and showed a good linearity in the range of 25-1 600 nmol/L. The LOD and LOQ were 0.5 nmol/L and 1.0 nmol/L, respectively. A cortisol assay kit (chemiluminescence immunoassay) traced to this candidate reference method was used to determine 46 clinical serum samples concurrently, and the two methods exhibited good correlation.Conclusions:A candidate reference method for the determination of cortisol in serum was established, demonstrating high sensitivity, good repeatability and accuracy. This method can serve as a reference for the measurement traceability and accuracy evaluation of routine methods.

Plant fungal pathogens secrete numerous proteins into the apoplast at the plant-fungus contact sites to facilitate colonization.However,only a few secretory proteins were functionally characterized in Magnaporthe oryzae,the fungal pathogen causing rice blast disease worldwide.Asparagine-linked glycosylation 3(Alg3)is an a-l,3-mannosyltransferase functioning in the N-glycan synthesis of N-glycosylated secretory proteins.Fungal pathogenicity and cell wall integrity are impaired in Aalg3 mutants,but the secreted proteins affected in Aalg3 mutants are largely unknown.In this study,we compared the secretomes of the wild-type strain and the Aalg3 mutant and identified 51 proteins that require Alg3 for proper secretion.These proteins were predicted to be involved in metabolic processes,interspecies interactions,cell wall organization,and response to chemicals.Nine proteins were selected for further validation.We found that these proteins were localized at the apoplastic region surrounding the fungal infection hyphae.Moreover,the N-glycosylation of these proteins was significantly changed in the Aalg3 mutant,leading to the decreased protein secretion and abnormal protein localization.Furthermore,we tested the biological functions of two genes,INV1(encoding invertase 1,a secreted invertase)and AMCase(encoding acid mammalian chinitase,a secreted chitinase).The fungal virulence was significantly reduced,and the cell wall integrity was altered in the Ainv1 and Aamcase mutant strains.Moreover,the N-glycosylation was essential for the function and secretion of AMCase.Taken together,our study provides new insight into the role of N-glycosylated secretory proteins in fungal virulence and cell wall integrity.