Evaluation of swallowing function after cricohyoidoepiglottopexy / 局解手术学杂志

Objective To investigate the evaluation ways and effects of swallowing function after cricohyoidoepiglottopexy(CHEP). Methods Selected 92 patients of glottic carcinoma who were admitted into hospital from February 2014 to January 2017,and all the patients were given cricohyoidoepiglottopexy(CHEP)therapy and function reconstruction.Modified barium swallow(MBS),modified penetration as-piration scale(MPAS),and fiberoptic endoscopic evaluation of swallowing(FEES)were applied after the surgery.And the prognosis of patients was followed up.Results There was one patient who was not able to extubate,and the extubation time of tracheaostomy tube and stomach tube were respectively(12.04 ±5.42)week and(8.00 ±2.19)d among the remaining 91 cases.Three months after operation,the laryngeal function were good in 84 cases,moderate in 6 cases and poor in 2 cases,the incidence of complications was 6.5%.The fundamental frequency and fundamental frequency perturbation three months after operation were significantly lower than thos before operation(P<0.05). With the extension of postoperative time,the MPAS score of patients with MBS and FEES evaluation were obviously decreased(P<0.05). The MBS assessment score were respectively(3.87 ±0.98)points,(1.64 ±0.65)points,(1.09 ±0.33)points at 15 days,30 days and 60 days after operation.The FEES evaluation score were respectively(3.27 ±1.33)points,(1.73 ±1.11)points,(1.18 ±0.89)points at 15 days,30 days and 60 days after operation.With the MBS assessment as the gold standard,the sensitivity of FEES assessment to normal,false aspiration and aspiration were 100%,76.7%and 86.7%,respectively,and the specificity were 86.7%,97.1% and 98.3%,respectively. Conclusion The cricohyoidoepiglottopexy and laryngeal defect repair in the treatment of glottic carcinoma can effectively preserve the laryn -geal function,reduce the incidence of postoperative complications,improve pronunciation function,and the FEES and MBS evaluation of laryn-geal function have good accuracy,and they have good clinical significance to understand the degree of postoperative aspiration.

Construction of biofilm formation related mutants in Vibrio parahaemolyticus / 中华预防医学杂志

<p><b>OBJECTIVE</b>To construct the mutants of biofilm related genes in Vibrio parahaemolyticus and confirm the mutants.</p><p><b>METHODS</b>The homologous upstream and downstream flanking fragments of target gene were amplified by using PCR, and the fusion homologous fragment was amplified by using the two flanking fragments as template. Then the fusion homologous fragment was digested by restriction enzyme and cloned into suicide plasmid pDS132. The recombinant plasmid was transferred into Vibrio parahaemolyticus RIMD 2210633 through conjugation. The mutants were screened and identified by PCR and the phenotype of one mutant was analyzed in order to verify that the mutants were constructed successfully.</p><p><b>RESULTS</b>Six recombinant plasmids carrying the fusion homologous fragments of genes vbfR, crp, hns, swrZ, swrT and cpsR respectively were constructed and identified by PCR. The amplification products of 1190, 1128, 1136, 953, 1242 and 1112 bp were obtained respectively. The six mutants (ΔvbfR, Δcrp, Δhns, ΔswrZ, ΔswrT and ΔcpsR) were constructed using recombinant plasmids. Verified by PCR, the size of amplification products of mutants (1190, 1128, 1136, 953, 1242 and 1112 bp respectively) was less (610, 739, 421, 542, 427 and 1367 bp respectively) than the corresponding positive control. Meanwhile, none of the products was amplified using the primers locating on the target gene. One mutant Δhns was selected to test the ability of biofilm formation. The result showed that the ability of biofilm formation of mutant Δhns was increased compared with the wild type.</p><p><b>CONCLUSION</b>Six mutants of biofilm related genes in Vibrio parahaemolyticus were constructed and tested by molecular and phenotype experiment to confirm that the mutants were constructed successfully.</p>

Establishment of stable subline of K562 cells expressing human leucocyte antigen a1101 / 中国实验血液学杂志

The aim of this study was to establish a stable subline of K562 cells expressing the HLA-A(*)1101 protein, which was expected to provide target cells for characterizing the HLA-I restrictive antigen specific cytotoxic T lymphocyte (CTL) effects against chronic myeloid leukemia (CML). The HLA-A(*)1101 protein encoding gene was amplified from peripheral blood mononuclear cell (PBMNC) of CML patient by RT-PCR; the 2A peptide linker (D-V-E-X-N-P-G-P) gene was linked to the 3'terminal of the HLA-A(*)1101 gene by recombinant PCR, then the recombinant was cloned into the pEGFP-N3 plasmid which contains an enhanced green fluorescent protein gene, and the eukaryotic recombinant expression vector containing HLA-A(*)1101-T2A-EGFP transcription box was constructed; the pEGFP-N3 vector and recombinant vector was separately electroporated into K562 cells. The expression of GFP was monitored by fluorescence microscopy, finally stably transfected sublines of K562 cells containing HLA-A(*)1101 gene, and of K562 containing pEGFP-N3 vector were obtained by G418 selection; the transcriptional or translational expression of HLA-A(*)1101 gene was detected with RT-PCR and flow cytometry respectively. The results indicated that the eukaryotic expression vector HLA-A(*)1101-T2A-EGFP plasmid was successfully constructed; after G418 selection for 2 months, two sublines of K562 cells (HLA-A(*)1101(+)K562, pEGFP-N3(+)K562) expressing GFP were constructed. The expression of HLA-A*A1101 gene could be determined in HLA-A(*)1101(+)K562 cell line by RT-PCR, while the pEGFP-N3(+)K562 cells could not express HLA-A*A1101 gene. HLA-A(*)1101 protein and GFP double positive HLA-A(*)1101(+)K562 cells were up to 88.5%, which was obviously higher than pEGFP-N3(+)K562 cells (0.698%) by flow cytometric analysis. It is concluded that a simple and effective method to select HLA-A(*)1101(+)K562 cells has been established and a subline of K562 cell expressing HLA-A(*)1101 protein on its cell membrane was successfully constructed, which provides the tool cells for further studying the specific cellular immunity against-CML.