OBJECTIVE To observe the characteristics of outer membrane vesicles(OMVs)of 1 strain of hypervir-ulent Klebsiella pneumoniae(hvKp)11492.METHODS The hvKp11492,one of major clones of hvKp that were i-solated from patients with bloodstream infection in the First Medical Center of Chinese PLA General Hospital,was chosen as the research subject.The strain was identified by means of matrix-assisted laser desorption/ioniza-tion time of flight mass spectrometry and whole genome sequencing.The hvKp1 1492-OMVs were separated and purified by high speed centrifugation in combination with polymer precipitation,the morphology and particle size of the hvKp11492-OMVs were identified by transmission electron microscopy(TEM)and nanoparticle tracking a-nalysis(NTA),the proteomic characteristics were analyzed through bicinchoninic acid assay(BCA)and liquid chromatography-mass spectrometry(LC-MS)series technique,and the virulence genes were detected by PCR.RESULTS The hvKp11492-OMVs were displayed differently in size,round or oval,and complete double-layer membrane vesicles on TEM.The result of NTA showed that the average particle size of the hvKp1 1492-OMVs was 270 nm.The protein content of hvKp11492-OMVs was(2.448±0.975)μg/μl.The result of subcellular lo-calization indicated that the protein included plasmosin,intracellular membrane protein,periplasmic protein,ex-tracellular membrane protein and nuclear region protein,which were found,by the annotation of GO database,to participate in the biological processes such as oxidation-reduction,interpretation,and metabolism.The hvKp11492-OMVs contained various proteins such as GDP-L-fucose synthetase,iron ion transporter protein and ferritin that were associated with pathogenicity.In addition,the hvKp1 1492-OMVs carried with iutA,iroN,iucA and rmpA virulence genes.CONCLUSIONS The morphologic characteristics,size and proteomic characteris-tics of the hvKp11492-OMVs are identified in the study.It is concluded that the hvKp11492-OMVs carry with va-rious proteins and genes that are association with the virulence and pathogenicity.

OBJECTIVE To evaluate the performance of commonly used molecular biological identification methods in differential identification of Citrobacter freundii complex(Cfc)and find out the differences in the mass spectra among the prevalent species of Cfc so as to facilitate the rapid and accurate identification.METHODS Totally 150 clinical strains were isolated from The First Medical Center of Chinese PLA General Hospital from 2015 to 2020 and were identified as Cfc by means of matrix assisted laser desorption ionization-time-of-flight mass spectrometry(MALDI-TOF MS),the genome data were obtained by the next generation sequencing and were compared with reference genome of Citrobacter species from National Center of Biotechnology Information(NCBI)based on aver-age nucleotide identity(ANI),phylogenetic analyses of rec N gene,concatenated alignments of four housekeeping genes(fusA,pyrG,leuS and rpoB as well as gyrB,rplB,fusA and leuS),core genome single nucleotide poly-morphism(coreSNP)and whole genome(WG),digital DNA-DNA hybridization(dDDH)analysis,ribosomal multilocus sequence typing(rMLST)and typical(strain)genome service(TYGS)analysis.ANI was set as the'gold standard' for the identification of the Cfc strains,and the capabilities of the various methods were evaluated.A certain number of strains of Citrobacter freundii(Cfr),Citrobacter braakii(Cbr)and Citrobacter portucalen-sis(Cpo)were randomly selected to obtain and compare the mass spectra so as to find out the differences in the mass spectra.RESULTS The identification capability of the Cfc strains is limited based on the available database for MALDI-TOF MS,Cpo was prone to be misidentified as Cfr or Cbr.The mass spectrum between Cpo and Cfr ranged between 2600 and 2660 m/z,the mass spectrum between Cpo and Cbr ranged between 5200 and 5300 m/z,showing significant difference.The concatenated alignments of four housekeeping genes gyrB,rplB,fusA and leuS,CoreSNP and WG phylogenetic analysis had high accurate rate of identification.CONCLUSIONS The meth-ods have certain limitations in identification of Cfc.The joint application of ANI with the methods such as WG phylogenetic analysis,update of mass spectrum database and timely establishment of database can achieve the ac-curate identification of the species of Cfc.

After continuous development and improvement, lung transplantation has become the preferred means to treat a variety of benign end-stage lung diseases. However, the field of lung transplantation still faces many challenges, including shortage of donor resources, preservation and maintenance of donor lungs, and postoperative complications. Lung tissue samples removed after lung transplantation are excellent clinical resources for the study of benign end-stage lung disease and perioperative complications of lung transplantation. However, at present, the collection, storage and utilization of tissue samples after lung transplantation are limited to a single study, and unified technical specifications have not been formed. Based on the construction plan of the biobank for lung transplantation in the First Affiliated Hospital of Xi'an Jiaotong University, this study reviewed the practical experience in the collection, storage and utilization of lung transplant tissue samples in the aspects of ethical review, staffing, collection process, storage method, quality control and efficient utilization, in order to provide references for lung transplant related research.

Objective:To analyze the correlation between thymus dose-volume parameters and lymphopenia in patients with early-stage breast cancer (BC) receiving adjuvant radiotherapy (RT).Methods:Medical records of 54 patients with early-stage BC who received postoperative adjuvant RT in the Second Affiliated Hospital of Soochow University from January to December 2019 were retrospectively analyzed. Absolute lymphocyte counts (ALC) were collected at 1 month before (baseline) and weekly during RT. Lymphopenia was graded based according to the common terminology criteria for adverse events version 5.0 and nadir/baseline ALC was calculated. The thymus was delineated according to anatomical boundaries in the original RT planning system. Dosimetric parameters were obtained from the dose volume histograms. Stepwise multiple linear regression analysis was used to explore the factors associated with nadir/baseline ALC. The cutoff values of dosimetric parameters for predicting ≥grade 3 lymphopenia were obtained using the receiver operating characteristic (ROC) curve.Results:The proportion of 54 patients experiencing ≥ grade 3 lymphopenia was 38.9%. The median value of thymus volume, mean dose, V 5 Gy, V 10 Gy were 14.02 cm 3, 4.95 Gy, 36.18%, and 6.61%, respectively. Stepwise multiple linear regression analysis revealed that baseline ALC ( P=0.005), quadrant location ( P=0.005) and mean thymus dose ( P<0.001) were significantly associated with nadir/baseline ALC. ROC curve analysis indicated that the cutoff values of thymus mean dose, V 5 Gy and V 10 Gy for predicting ≥ grade 3 lymphopenia were 6.12 Gy, 35.2%, and 7.4%, respectively. Conclusions:Lymphopenia in early-stage BC patients is significantly correlated with high dosimetric parameters of the thymus during postoperative adjuvant RT. Thymus may be considered as an organ at risk during RT.

OBJECTIVE To observe the characteristics of outer membrane vesicles(OMVs)of 1 strain of hypervir-ulent Klebsiella pneumoniae(hvKp)11492.METHODS The hvKp11492,one of major clones of hvKp that were i-solated from patients with bloodstream infection in the First Medical Center of Chinese PLA General Hospital,was chosen as the research subject.The strain was identified by means of matrix-assisted laser desorption/ioniza-tion time of flight mass spectrometry and whole genome sequencing.The hvKp1 1492-OMVs were separated and purified by high speed centrifugation in combination with polymer precipitation,the morphology and particle size of the hvKp11492-OMVs were identified by transmission electron microscopy(TEM)and nanoparticle tracking a-nalysis(NTA),the proteomic characteristics were analyzed through bicinchoninic acid assay(BCA)and liquid chromatography-mass spectrometry(LC-MS)series technique,and the virulence genes were detected by PCR.RESULTS The hvKp11492-OMVs were displayed differently in size,round or oval,and complete double-layer membrane vesicles on TEM.The result of NTA showed that the average particle size of the hvKp1 1492-OMVs was 270 nm.The protein content of hvKp11492-OMVs was(2.448±0.975)μg/μl.The result of subcellular lo-calization indicated that the protein included plasmosin,intracellular membrane protein,periplasmic protein,ex-tracellular membrane protein and nuclear region protein,which were found,by the annotation of GO database,to participate in the biological processes such as oxidation-reduction,interpretation,and metabolism.The hvKp11492-OMVs contained various proteins such as GDP-L-fucose synthetase,iron ion transporter protein and ferritin that were associated with pathogenicity.In addition,the hvKp1 1492-OMVs carried with iutA,iroN,iucA and rmpA virulence genes.CONCLUSIONS The morphologic characteristics,size and proteomic characteris-tics of the hvKp11492-OMVs are identified in the study.It is concluded that the hvKp11492-OMVs carry with va-rious proteins and genes that are association with the virulence and pathogenicity.

Objective:To analyze the correlation between thymus dose-volume parameters and lymphopenia in patients with early-stage breast cancer (BC) receiving adjuvant radiotherapy (RT).Methods:Medical records of 54 patients with early-stage BC who received postoperative adjuvant RT in the Second Affiliated Hospital of Soochow University from January to December 2019 were retrospectively analyzed. Absolute lymphocyte counts (ALC) were collected at 1 month before (baseline) and weekly during RT. Lymphopenia was graded based according to the common terminology criteria for adverse events version 5.0 and nadir/baseline ALC was calculated. The thymus was delineated according to anatomical boundaries in the original RT planning system. Dosimetric parameters were obtained from the dose volume histograms. Stepwise multiple linear regression analysis was used to explore the factors associated with nadir/baseline ALC. The cutoff values of dosimetric parameters for predicting ≥grade 3 lymphopenia were obtained using the receiver operating characteristic (ROC) curve.Results:The proportion of 54 patients experiencing ≥ grade 3 lymphopenia was 38.9%. The median value of thymus volume, mean dose, V 5 Gy, V 10 Gy were 14.02 cm 3, 4.95 Gy, 36.18%, and 6.61%, respectively. Stepwise multiple linear regression analysis revealed that baseline ALC ( P=0.005), quadrant location ( P=0.005) and mean thymus dose ( P<0.001) were significantly associated with nadir/baseline ALC. ROC curve analysis indicated that the cutoff values of thymus mean dose, V 5 Gy and V 10 Gy for predicting ≥ grade 3 lymphopenia were 6.12 Gy, 35.2%, and 7.4%, respectively. Conclusions:Lymphopenia in early-stage BC patients is significantly correlated with high dosimetric parameters of the thymus during postoperative adjuvant RT. Thymus may be considered as an organ at risk during RT.

OBJECTIVE To evaluate the performance of commonly used molecular biological identification methods in differential identification of Citrobacter freundii complex(Cfc)and find out the differences in the mass spectra among the prevalent species of Cfc so as to facilitate the rapid and accurate identification.METHODS Totally 150 clinical strains were isolated from The First Medical Center of Chinese PLA General Hospital from 2015 to 2020 and were identified as Cfc by means of matrix assisted laser desorption ionization-time-of-flight mass spectrometry(MALDI-TOF MS),the genome data were obtained by the next generation sequencing and were compared with reference genome of Citrobacter species from National Center of Biotechnology Information(NCBI)based on aver-age nucleotide identity(ANI),phylogenetic analyses of rec N gene,concatenated alignments of four housekeeping genes(fusA,pyrG,leuS and rpoB as well as gyrB,rplB,fusA and leuS),core genome single nucleotide poly-morphism(coreSNP)and whole genome(WG),digital DNA-DNA hybridization(dDDH)analysis,ribosomal multilocus sequence typing(rMLST)and typical(strain)genome service(TYGS)analysis.ANI was set as the'gold standard' for the identification of the Cfc strains,and the capabilities of the various methods were evaluated.A certain number of strains of Citrobacter freundii(Cfr),Citrobacter braakii(Cbr)and Citrobacter portucalen-sis(Cpo)were randomly selected to obtain and compare the mass spectra so as to find out the differences in the mass spectra.RESULTS The identification capability of the Cfc strains is limited based on the available database for MALDI-TOF MS,Cpo was prone to be misidentified as Cfr or Cbr.The mass spectrum between Cpo and Cfr ranged between 2600 and 2660 m/z,the mass spectrum between Cpo and Cbr ranged between 5200 and 5300 m/z,showing significant difference.The concatenated alignments of four housekeeping genes gyrB,rplB,fusA and leuS,CoreSNP and WG phylogenetic analysis had high accurate rate of identification.CONCLUSIONS The meth-ods have certain limitations in identification of Cfc.The joint application of ANI with the methods such as WG phylogenetic analysis,update of mass spectrum database and timely establishment of database can achieve the ac-curate identification of the species of Cfc.

To explore the application of humanistic care in children's fever clinics,the children's fever clinics of Guangzhou Women and Children's Medical Center conducted relevant research using narrative medicine as a guiding concept.This paper elaborated on the medical humanistic dilemmas and narrative care needs in children's fever clinics,and focused on exploring the practice paths of narrative medicine in pediatric with Chinese characteristics from five dimensions,including the cultivation of nursing staff's abilities of professional narrative and humanistic care,the establishment of health lectures featuring narrative patient education,the alleviation of medical anxiety for children and their families,related support of narrative nursing,and caring services.The aim was to improve the narrative care ability of nurses in children's fever clinics,develop methods for pediatric patients that can eliminate the fear of seeking medical treatment,and protect children's physical and mental health.A carrier of care,support,and emotional expression can be provided for parents.The foundation for nursing staff to achieve professional growth through narrative reflection can be laid.Thus,it can assist in establishing a life narrative community relationship between doctors and patients,and jointly explore the meaning of life.

Continuous arterial blood pressure monitoring by radial artery catheterization is an indispensable monitoring technique for children in the perioperative period and critical care, and the factors that affect the success of catheterization have always been the focus of clinical researches. This study reviewed the research progress on the factors related to the success of radial artery catheterization in children from the perspectives of operator's factors, patient's factors, and operation equipment and materials.

Closed-loop hospital management can effectivly cope with the COVID-19 pandemic. In order to ensure the continuity of treatments for hemodialysis patients under closed-loop management and minimize possible medical and infection risks, Huashan Hospital affiliated to Fudan University and 9 hospitals in Shanghai established a hemodialysis alliance in January 2021.The alliance optimized hemodialysis resources within the region through overall planning by preparing sites, materials and personnel shifts in advance, and establishing management systems and work processes to ensure that patients could be quickly and orderly diverted to other blood dialysis centers for uninterrupted high-quality hemodialysis services, in case that some hemodialysis centers in the alliance under closed-loop management.From November 2021 to April 2022, 317 of 1 459 hemodialysis patients in the alliance were diverted to other centers for treatment, accumulating 1 215 times/cases of treatments without obvious adverse reactions. The practice could provide a reference for medical institutions to quickly establish mutual support mode under major public health events.