Development of ELISAs for the detection of urogenital chlamydia trachomatis infection targeting the pORF5 protein / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To prepare antibodies against pORF5 plasmid protein of Chlamydia trachomatis and develop double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISAs) for the detection of genital C. trachomatis infections.</p><p><b>METHODS</b>The pORF5 protein was expressed in Escherichia coli and used to immunize BALB/c mice and New Zealand rabbits to produce monoclonal antibodies (mAbs) and polyclonal antibody (pAb) for DAS-ELISAs. Clinical samples from 186 urogenital infection patients (groups I) and 62 healthy donors (groups II) were detected in parallel by the DAS-ELISAs developed in this study and by IDEIA PCE commercial ELISA.</p><p><b>RESULTS</b>Two hybridoma cell lines, named 2H4 and 4E6, stably secreting specific mAbs against pORF5 were obtained. The mAb 2H4 was recognized by 32 (17.20%, positive recognition rate) and 25 (13.44%), mAb 2H4 by 0 (0%) and 2 (3.22%) samples from groups I and II, respectively. The sensitivities of mAbs 2H4 and 4E6 were 92.11% and 77.78% and the specificities were 100% and 96.88%, respectively in relation to the IDEIA PCE commercial ELISA. The sensitivities of detection for the DAS-ELISAs were 10 ng/mL (based on 2H4) and 18 ng/mL (based on 4E6).</p><p><b>CONCLUSION</b>Two DAS-ELISAs were developed in this study that provided a feasible and effective assay that could be considered alternative tools for the serodiagnosis of C. trachomatis infection.</p>

Predictive value of three preoperative nutritional assessment methods in postoperative complications of gastrointestinal cancers / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To compare the value of three preoperative nutritional assessment methods, European nutrition risk screening 2002(NRS 2002), mini-nutrition assessment(MNA) and subjective global assessment(SGA), in predicting postoperative complications of gastrointestinal cancer patients.</p><p><b>METHODS</b>A total of 235 patients with gastrointestinal cancers, including 31 esophageal cancers, 82 gastric cancers, and 122 colorectal cancers, in our hospital from January 2012 to June 2013 were prospectively enrolled. Preoperative nutritional status was evaluated with above 3 methods respectively. Postoperative complication rates were compared among different preoperative nutritional status.</p><p><b>RESULTS</b>According to SGA score, the morbidity of severe-moderate, mild and no malnourished patients was 40.5%(17/42), 25.3%(22/87) and 14.2%(15/106) respectively(P<0.01). According to MNA score, the morbidity of patients with malnutrition, at risk of malnutrition and without malnutrition was 32.9%(23/70), 24.7%(18/73) and 14.1%(13/92) respectively(P<0.05). According to NRS 2002, the morbidity of patients at malnutrition risk and without malnutrition risk was 27.6%(27/98) and 19.7%(27/137) respectively(P>0.05). Multiple regression analysis revealed that both SGA and MNA scores were predictive factors for the development of postoperative complications(both P<0.01). The sensitivity of SGA score for predicting complications was higher compared to MNA score (90.7% vs. 79.6%), while the specificity was similar(49.7% vs. 50.8%).</p><p><b>CONCLUSIONS</b>Both SGA and MNA scores can effectively predict the development of postoperative complications in gastrointestinal cancer patients, and SGA score has better sensitivity. SGA score is recommended for decision-making regarding preoperative nutrition support.</p>

Cloning and cellular localization of pORF8 plasmid protein of Chlamydia trachomatis / 南方医科大学学报

<p><b>OBJECTIVE</b>To clone the plasmid protein pORF8 of Chlamydia trachomatis and localize its expression in Chlamydia-infected cells.</p><p><b>METHODS</b>pORF8 gene was amplified and cloned into pGEX-6p vector, and the pORF8 fusion protein was expressed in E.coli XL1 Blue. After purification with glutathione-conjugated agarose beads, the pORF8 fusion protein was used to immunize BALB/c mice to generate polyclonal antibodies against pORF8 protein. The antibodies obtained were used to localize the plasmid protein pORF8 in Chlamydia-infected cells with immunofluorescence assay (IFA).</p><p><b>RESULTS</b>The pORF8 gene 744 bp in length was successfully cloned and the GST fusion protein with a relative molecular mass of 54 000 was obtained. The cellular distribution pattern of the plasmid protein pORF8 was similar to that of the major outer membrane protein (MOMP), a known C. trachomatis inclusion body protein, but not to that of chlamydial protease-like activity factor (CPAF, a secreted protein).</p><p><b>CONCLUSION</b>The plasmid protein pORF8 is localized on the bacterial organism as an inclusion body protein in C. trachomatis-infected cells. The cellular location of pORF8 protein can potentially provide important insights into the pathogenesis of C. trachomatis.</p>