ObjectiveTo explore the effects of enteral nutrition (EN) on intestinal permeability in patients with active ulcerative colitis (AUC). MethodsTwenty-four A UC patients were randomly divided into two groups:routine treatment group (n =11 ) and routine treatment plus EN group (n =13). Patients in routine treatment group were treated with mesalazine as well as low-residue diet, while patients in routine treatment plus EN group received mesalazine and short peptide EN for 14 days. The ratio of lactulose to mannitol in urine (L/M) before and after treatment was detected by high-performance liquid chromatography. ResultsThe L/M ratio was 0. 039 ± 0. 025 in routine treatment group and 0.072 ± 0.019 in routine treatment plus EN group (P =0.069). After 2 weeks of treatment, the L/M ratio of routine treatment plus EN group (0.038 ± 0.012 ) was significantly lower than the pretreatment level (P =0.043 ), while the L/M ratio of routine treatment group between before and after treatment had no significant difference (0.039 ± 0.025 vs. 0.032 ± 0.022, P =0.730). ConclusionEN can effectively improve the intestinal permeability in AUC patients.

Rv2742 is a novel gene identified from Mycobacterium tuberculosis H37Rv by the proteogenomics strategy. The aim of this study was to establish a system of soluble expression and purification of the missing protein Rv2742 in M. tuberculosis H37Rv, to provide reference for further research on the biological function of Rv2742. The soluble protein was not successfully induced by prokaryotic expression vectors pGEX-4T-2-Rv2742, pET-32a-Rv2742, pET-28a-Rv2742 and pMAL-c2X-Rv2742. After the codon of novel gene Rv2742 was optimized according to E. coli codon usage frequency, only the recombinant strain containing plasmid pMAL-c2X-Rv2742 could produce soluble products of Rv2742 encoding gene. In addition, the expression effects of the desired fusion protein were also analyzed under different conditions including hosts, culture temperatures and IPTG concentrations. The optimum expression conditions were as follows: Rosetta (DE3) host, 16 °C culture temperature and 0.5 mmol/L IPTG. After being purified by affinity chromatography with amylose resin, the fusion protein sequence was confirmed by LC-MS/MS. These results indicated that the novel gene Rv2742 product could be successfully induced and expressed in a soluble form by the expression system pMAL-c2X with MBP tag. Our findings provide reference for studies on potential interaction and immunogenicity.
Chromatography, Liquid ; Cloning, Molecular ; Escherichia coli ; Mycobacterium tuberculosis ; genetics ; Recombinant Fusion Proteins ; Tandem Mass Spectrometry

Type 2 diabetes mellitus (T2DM) is a global metabolic disease with potentially life- threatening complications. Liver metabolism plays a vital role in the occurrence and development of diabetes mellitus. It has been reported that the Chinese medicinal Anemarrhenae rhizoma (AR) can relieve insulin resistance and diabetes mellitus. However, the effect on abnormal liver metabolism in diabetes mellitus is still unclear. Therefore, we extracted liver proteins of T2DM rats induced by high-fat diet (HFD) and streptozotocin (STZ), T2DM rats treated with AR extract (ARE), obesity rats (fed with HFD), and normal control rats (fed with normal diet). Then, through tandem mass tag (TMT) labeling combined with mass spectrometry (MS), we obtained the quantitative proteomic data. Bioinformatics software was used for hierarchical cluster analysis and principal component analysis of the data in each group. The volcano map for differentially expressed proteins (P < 0.05, fold change > 1.5) was plotted. It was found that the treatment group was closer to the normal control group, indicating that the quantitative proteomic data of liver tissue can reflect the therapeutic effect of ARE on T2DM rats. Key protein clusters closely related to the treatment of ARE were screened out. The Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of the protein clusters were analyzed by David, and the result showed that AR's alleviation of abnormal fatty acid metabolism in livers of T2DM rats may be related to the regulation of the expression of key proteins Ndufa6 and Prkar2b.
Rats ; Animals ; Diabetes Mellitus, Type 2/metabolism* ; Proteomics/methods* ; Drugs, Chinese Herbal/therapeutic use* ; Anemarrhena ; Liver/metabolism*