Objective To evaluate our results of external quality assessment (EQA) program of the newborn screening laboratory for 6 years, summarize the problems in the screening measurement and improve the quality of measurement.Methods The phenylalanine (Phe) and thyrotropin (TSH) values in dried blood spot specimens on filter paper distributed by national center were measured by fluorometric method and time -resolved fluorescence im-munoassay ( DELFIA ) respectively.The results sent by national center were analyzed .Results In the 90 specimens of 18 batches, the bias range of Phe and TSH measurement was wide (-62.50%~66.67%and-66.67%~54.31%respectively) and their average values were 8.62%and 10.18% respectively;85 quantitative results of Phe and 84 quantitative results of TSH measurement were in the quality control , accounting for 94.44%and 93.33%respective-ly.The qualitative analysis of 90 Phe and TSH specimens all conformed to the expected results , accounting for 100%.The result of every batch was up to 80%.Conclusion Our neonatal screening laboratory is in control and it is useful to evaluate the measurement competence , find out the problems in the Phe and TSH measurement and re-solve them in time through participated in the national EQA in neonatal screening laboratory , so as to improve the quality of measurement and ensure the accuracy and reliability of results .

In vitro evolution methods are often used to modify protein with improved characteristics. We developed a directed evolution protocol to enhance the thermostability of the beta-1,3-1,4-glucanase. The thermostability of the enzyme was significantly improved after two rounds of directed evolution. Three variants with higher thermostability were obtained. The mutant enzymes were further analyzed by their melting temperature, halftime and kinetic parameters. Comparing to intact enzyme, the T50 of mutant enzymes 2-JF-01, 2-JF-02 and 2-JF-03 were increased by 2.2 degrees C, 5.5 degrees C and 3.5 degrees C, respectively, the halftime (t1/2, 60 degrees C) of mutant enzymes 2-JF-01, 2-JF-02 and 2-JF-03 were shortened by 4,13 and 17 min, respectively, the V(max) of mutant enzymes were decreased by 8.3%, 2.6% and 10.6%, respectively, while K(m) of mutant enzymes were nearly unchanged. Sequence analysis revealed seven single amino acid mutant happened among three mutant enzymes, such as 2-JF-01 (N36S, G213R), 2-JF-02 (C86R, S115I, N150G) and 2-JF-03 (E156V, K105R). Homology-modeling showed that five of seven substituted amino acids were located on the surface of or in hole of protein. 42.8% of substituted amino acids were arginine, which indicated that arginine may play a role in the improvement of the thermostability of the beta-1,3-1,4-glucanase.This study provide some intresting results of the structural basis of the thermostability of beta-1,3-1,4-glucanase,and provide some new point of view in modifying enzyme for future industrial use.
Amino Acid Sequence ; Bacillus ; enzymology ; Directed Molecular Evolution ; Endo-1,3(4)-beta-Glucanase ; biosynthesis ; chemistry ; genetics ; Enzyme Stability ; Hot Temperature ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; genetics

Brewer's yeast is crucial in beer fermentation, mainly beer flavor diversity and stability. Beer flavor stability is one of the most influential beer quality aspects, and screening or breeding brewer's yeast with enhanced anti-staling capacity will be an effective solution. In recent decades, with the progress of genetic engineering and detailed description of brewer's yeast genome, great efforts have been made to improve brewer's yeast. This review highlights recent advances in classical and genetic engineering improvement of yeasts to produce more antioxidant compounds or less beer aging substances and precursors. Therein, improvement targets, evaluation methods and development strategies of anti-staling brewer's yeast are also discussed. Furthermore, hotspot and future trend of anti-staling yeast strain development are also proposed.

[Abstract] Objective: To explore the pathogenosis and prognostic markers for non-smoking female lung cancer patients with bioinfor‐ matics analysis and functional prediction of potential lung cancer associated genes in female non-smokers. Methods: Data for nonsmoking female patients with lung cancer were downloaded from the Gene Expression Omnibus (GEO) database and the differentially expressed genes (DEGs) were identified using GEO2R. DAVID online data base was used to perform gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG), and STRING online software was used to perform protein-protein interaction (PPI) analysis; then the plug-in (M-CODE) was used to screen the key DEGs; finally, GEPIA and Kaplan-Meier plotter were used to perform function prediction and prognosis analysis of key DEGs. Results: A total of 160 DEGs were screened, including 54 up-regulated and 106 down-regulated genes; GO enrichment analysis showed that these DEGs were mainly related to neovascularization, single cell adhesion, positive regulation of GTPase activity and signal transduction (all P<0.05). KEGG pathway analysis revealed that DEGs were mainly involved in cell adhesion molecules (CAMs), leukocyte transendothelial migration, tight junction and endocytosis (all P<0.05); PPI network analysis revealed 8 key DEGs, including TIE1, PECAM1, CLDN5, VEGFD, ICAM2, ESAM, EMCN and ROBO4. Conclusion: TIE1, CLDN5, ICAM2, ESAM, VEGFD and ROBO4 may be the research targets of the pathogenesis of non-smoking female lung cancer patients. PECAM1 and EMCN may be the new bio-markers to predict the progression and prognosis of nonsmoking female lung cancer patients.

Yeast cell wall plays an important role in the establishment and maintenance of cell morphology upon the cell wall stress. The cell wall of yeast consists of β-glucans, mannoproteins and chitin. The composition and structure remodel due to cell wall stress. Brewer's yeast cell wall exhibits stress response during long-term acclimation in order to adapt to environmental changes. This paper reviews the composition and structure of yeast cell wall and the molecular mechanisms of cell wall remodeling and signal pathway regulation.
Cell Wall ; Chitin ; Saccharomyces cerevisiae

The morphological structure and physiological indexes of Saccharomyces cerevisiae have changed during serial re-pitching due to the stress conditions in serial handlings and the cells become aging. It is of great significance to study the physiological changes of S. cerevisiae during serial re-pitching to understand the anti-aging effect of S. cerevisiae. In this paper, the changes of the physiological indexes during re-pitching of yeast are summarized, and based on the analysis of the previous works further research directions are proposed.

1,3-1,4-β-glucanase (E.C.3.2.1.73) is an important industrial enzyme which cleave β-glucans into oligosaccharides through strictly cutting the β-1,4 glycosidic bonds in 3-O-substituted glucopyranose units. Microbial 1,3-1,4-β-glucanase belongs to retaining glycosyl hydrolases of family 16 with a jellyroll β-sandwich fold structure. The present paper reviews the industrial application and protein engineering of microbial β-glucanases in the last decades and forecasts the research prospects of microbial β-glucanases.
Amino Acid Sequence ; Glycoside Hydrolases ; Models, Molecular ; Protein Engineering ; Substrate Specificity