Meta-analysis of the effect of hollow nail and bone plate on Lisfranc injury / 中国骨伤

OBJECTIVE@#To compare the clinical efficacy of screw and bone plate internal fixation in the treatment of Lisfranc injury.@*METHODS@#The databases of Wanfang, CNKI, Pubmed, EMBASE, VIP, BIOSIS and other databases were retrieved by computer, and the clinical trial literature from January 1, 2000 to August 1, 2021 was retrieved, the methodological quality of the included studies was strictly evaluated and the data were extracted, and the obtained data were meta-analyzed by Revman 5.4 software.@*RESULTS@#Nine randomized controlled trial literature and 10 retrospective cohort studies were included, of which 416 patients in the experimental group were treated with screw internal fixation, and 435 patients in the control group were treated with bone plate internal fixation. Meta-analysis showed that the surgical time of the bone plate internal fixation group was longer than that of the screw internal fixation group [MD=-14.40, 95%CI(-17.21, -11.60), P<0.000 01], the postoperative X-ray anatomical reduction of the bone plate internal fixation group [MD=0.47, 95%CI(0.25, 0.86), P=0.01], the excellent and good rate of postoperative American orthopedic foot and ankle society(AOFAS) foot function score[MD=0.25, 95%CI(0.15, 0.42), P<0.000 01], postoperative AOFAS foot function score [MD=-5.51, 95%CI(-10.10, -0.92), P=0.02] of the bone plate fixation group was better than those of the screw internal fixation group. Two kinds of operation method had no statistical different for postoperative fracture healing time[MD=1.91, 95%CI(-1.36, 5.18), P=0.25], postoperative visual analgue scale(VAS)[MD=0.38, 95%CI(0.09, 0.86), P=0.11], postoperative complications [MD=1.32, 95%CI(0.73, 2.40), P=0.36], the postoperative infection [MD=0.84, 95%CI(0.48, 1.46), P=0.53], the postoperative fracture internal fixation loosening [MD=1.25, 95% CI(0.61, 2.53), P=0.54], the postoperative incidence of traumatic arthritis [MD=1.80, 95%CI(0.83, 3.91), P=0.14].@*CONCLUSION@#Bone plate fixation has better short-term and medium-term results and lower reoperation rate in the treatment of Lisfranc injury, so it is recommended to use bone plate fixation in the treatment of Lisfranc injury.

Construction of the eukaryotic expression plasmid containing lvgA gene flanked with CpG motifs of Legionella pneumophila and its expression in NIH3T3 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the eukaryotic expression plasmid containing lvgA gene flanked with CpG motifs of Legionella pneumophila for its expression in NIH3T3 cells.</p><p><b>METHODS</b>lvgA gene flanked with CpG motifs of Legionella pneumophila was amplified by PCR. The PCR products was inserted into the eukaryotic expression plasmid pcDNA3.1/myc-his(+) to construct the recombinant plasmid pclvgA/CpG, which was subsequently transfected into NIH3T3 cells via lipofection. Immunofluorescence analysis was carried out to detect the transient expression of the plasmid in the cells.</p><p><b>RESULTS</b>Sequence analysis showed that the recombinant plasmid pclvgA/CpG contained the lvgA/CpG fragment with a length of 657 bp, encoding a protein of 27.7 Ku. Immunofluorescence analysis identified the transient expression of the recombinant plasmid pclvgA/CpG in NIH3T3 cells.</p><p><b>CONCLUSION</b>The lvgA gene flanked with CpG motifs of Legionella pneumophila has been constructed successfully, and the transient expression of the recombinant plasmid pclvgA/CpG can be detected in NIH3T3 cells.</p>

Legionella pneumophila lvgA and Hsp60 gene splicing and the fusion gene expression in E.coli / 南方医科大学学报

<p><b>OBJECTIVE</b>To fuse Legionella virulence gene (lvgA) with heat shock protein 60 gene (Hsp60) by PCR and detect the fusion gene expression in E.coli.</p><p><b>METHODS</b>The fragments of lvgA and Hsp60 genes having matching sequences at their ends to be fused were amplified from the genomic DNA of Legionella pneumophila by PCR, and the PCR products were mixed, denatured, reannealed, so that the strands with matching sequences at their 3' ends overlapped to serve as primers for each other. Extension of this overlap by DNA polymerase produced recombinant products. After amplification with outer primers, sufficient product of the fusion gene was harvested, which was inserted into the prokaryotic expression vector pGEX-4T-1 to construct the prokaryotic expression recombinant plasmid. After identification with restriction enzyme analysis, polymerase chain reaction and nucleotide sequence analysis, the E.coli BL21 containing the recombinant plasmid pGlvgA/Hsp60 was induced with IPTG and the expression of lvgA/Hsp60 was detected by SDS-PAGE and Western blot analysis.</p><p><b>RESULTS</b>The lvgA/Hsp60 fusion gene of 2,292 bp was amplified and the recombinant plasmid pGlvgA/Hsp60 was constructed successfully. A 117-kD GST-lvgA-Hsp60 fusion protein was detected in the E.coli containing the recombinant plasmid.</p><p><b>CONCLUSION</b>The recombinant plasmid for Legionella pneumophila lvgA/Hsp60 fusion gene is constructed successfully and this fusion protein can be expressed in prokaryotic cells efficiently, which make possible the immunological characterization of this fusion gene.</p>

Construction of a prokaryotic expression vector carrying mompS-linker-flaA fusion gene and its expression in E.coli / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the fusion expression vector of Legionella pneumophila mompS and flaA genes linked with a flexible chain for expression in E.coli.</p><p><b>METHODS</b>The flaA gene, an flagellum subunit gene of Legionella pneumophila, and mompS gene that encodes an major outer membrane protein of Legionella pneumophila, were amplified from the DNA of Legionella pneumophila by PCR and cloned into the prokaryotic expression vector pET32a (+) containing thioredoxin gene Trx. Following analysis of the recombinant plasmid (pET-LpSLF) with restriction endonuclease digestion, PCR and DNA sequencing, the expression of pET-LpSLF was induced with IPTG and the expressed fusion protein Trx-MOMPS-FlaA was examined with SDS-PAGE and Western blotting.</p><p><b>RESULTS</b>The results of restriction endonuclease digestion, PCR and DNA sequencing analysis showed that the flaA gene (1 440 bp) and the mompS gene (906 bp) were successfully amplified from Legionella pneumophila DNA, and the recombinant plasmid pET-LpSLF was constructed and expressed in E.coli as demonstrated by SDS-PAGE and Western blotting.</p><p><b>CONCLUSION</b>The fusion expression vector of mompS and flaA genes linked with a flexible chain has been successfully constructed and allows efficient expression of mompS-linker-flaA gene in E.coli, which enables further study of the immunogenicity and immunoprotection of Legionella pneumophila.</p>