[Quality assessment of the clinical practice guideline for diagnosis, treatment and prevention of pulmonary thromboembolism, 2018].

Objective: To assess the quality of the clinical practice guideline for diagnosis, treatment and prevention of pulmonary thromboembolism, 2018 in China, providing the references for updating and developing clinical practice guidelines of this field in the future. Methods: The quality of the clinical practice guideline for diagnosis, treatment and prevention of pulmonary thromboembolism, 2018 in China was assessed using the internationally recognized instrument Appraisal of Guidelines for Research and Evaluation Ⅱ (AGREE Ⅱ). AGREE Ⅱ instrument consisted of 23 items in six domains, followed by two overall assessment items. Each item was scored from 1 to 7. The final overall guideline quality considered all domain items. Results: The scores of the six AGREE Ⅱ domains were: Scope and purpose 76.4%, Stakeholder involvement 55.6%, Rigor of development 78.1%, Clarity and presentation 83.3%, Applicability 55.2%, and Editorial independence 66.7%. The guideline was recommended for clinical use. Among the 101 recommendations, recommendations based on Levels High, Moderate and Low evidence accounted for 7 (6.9%), 31 (30.7%) and 63 (62.4%), respectively. Conclusion: The methodological quality of the clinical practice guideline for diagnosis, treatment and prevention of pulmonary thromboembolism, 2018 in China was great, but the levels of evidence were not high. More efforts were urgently required to improve in Stakeholder involvement and applicability. Especially corresponding economic research evidence, as well as preferences of patients and the public should be considered in the future development of clinical practice guidelines.

Identification of O-antigen polymerase transcription and translation start signals and visualization of the protein in Salmonella enterica serovar Typhimurium.

The wzy/rfc gene, encoding the O-antigen polymerase, of Salmonella enterica serovar Typhimurium has been previously cloned and sequenced. In the present work, the wzy transcriptional startpoint was initially identified by primer extension. Next, wzy promoter strength in Escherichia coli K-12 was measured, and was found to be greater than that of the induced lac promoter. To define the Wzy translational startpoint, DNA including the wzy promoter and the putative first five residues of the Wzy protein was fused to the N-terminus of glutathione-S-transferase, and the fusion protein purified by affinity chromatography. N-terminal amino acid sequencing yielded the Wzy translational startpoint. Next, the Wzy protein was C-terminally tagged with the FLAG peptide, and immunoblotting of an S. typhimurium strain expressing a low-copy wzy-FLAG gene (five copies per cell) localized the intact Wzy protein in the cytoplasmic membrane of S. typhimurium cells. The Wzy protein was not well-expressed from a multi-copy wzy-FLAG+ plasmid in S. typhimurium, or in E. coli K-12.

Purification and characterization of a Hodgkin's and B cell-associated glycoprotein.

A glycoprotein (BLA.36), expressed on the plasma membrane of Hodgkin's cells and also on normal and malignant B lymphocytes and histiocytes, was identified by reaction with a monoclonal antibody. BLA.36 was not detectable on other hematopoietic, carcinoma or melanoma cell lines. BLA.36 was purified to homogeneity by extracting proteins from a Hodgkin's cell line (HDLM-3), followed by immunoaffinity chromatography, utilizing immobilized anti-BLA.36 antibody, and gel filtration on Sephacryl S-100 in the presence of protein dissociating agents. The purified component yielded a single band on sodium dodecyl sulphate/polyacrylamide gel electrophoresis under both non-reducing and reducing conditions, and closely related three isotypes of similar molecular weight and with the apparent isoelectric points that ranged from 5.0 to 5.2 on two-dimensional gel electrophoresis. The purified BLA.36 reacted with the original specific antibody, on both one- or two-dimensional gel electrophoresis, suggesting that antigenic determinant was not adversely affected during purification procedure. Competitive immunoprecipitation analyses and the determination of N-terminus sequence of the first 13 amino acid residues suggest that BLA.36 is unrelated to other known Hodgkin's or hematopoietic cell antigens. Finally, significance of BLA.36 expression on the growth of BLA.36-positive cell lines was studied. Blocking of BLA.36 with anti-BLA.36 antibody led to the in vitro growth-inhibition of BLA.36-positive cell lines. The antibody pre-absorbed with the purified BLA.36 was unable to exert growth-inhibition, demonstrating the specificity of reaction. In addition, the treatment of the BLA.36-positive cell lines with differentiation-inducing agent, alpha-interferon (alpha-IFN), down-regulated BLA.36 expression and also showed in vitro growth-inhibition of BLA.36-positive cell lines. Taken together, these results suggest a growth-related function of BLA.36.