Objective:To explore the clinical value of synovial fluid calprotectin for the diagnosis of periprosthetic joint infection (PJI).Methods:Based on prospective cohort study design, a total of 82 patients suspected of PJI after hip and knee arthroplasty in the First Medical Center of the PLA General Hospital from July 2021 to June 2022 were selected. Patients were divided into infection group (PJI, n=39) and non-infection group (non-PJI, n=43) according to the diagnostic criteria proposed by the Second International Consensus Conference in 2018. The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used for double-blind detection of calprotectin and internal reference standard (IRS) in synovial fluid of patients. The peaks of target protein and IRS were recorded for further analysis. Mann-Whitney U test was used to compare the concentrations of S100A8 and S100A9 between the two groups, and receiver operating characteristic curve (ROC) was used to analyze the diagnostic efficacy of S100A8 and S100A9 for PJI. Results:Calprotectin was detected as monomers S100A8 and S100A9. Synovial fluid S100A8 was significantly higher in the PJI group than that in the non-PJI group [1.57 (0.48, 4.17) vs 0.00 (0.00, 0.05), Z=?7.221, P<0.05]. Synovial fluid S100A9 was also significantly higher in the PJI group than that in the non-PJI group [0.74 (0.29, 1.70) vs 0.06 (0.00, 0.10), Z=?6.255, P<0.05]. When using S100A8 and S100A9 to diagnose PJI, the sensitivity were 97.4% and 87.2%, the specificity were 86.0% and 88.4%, and the area under the ROC were 0.964 (95% CI 0.929-0.998) and 0.902 (95% CI 0.924-0.996), respectively. Conclusion:The detection of synovial fluid S100A8 and S100A9 by MALDI-TOF MS can make a satisfactory diagnosis for PJI.

Objective:Based on the high-throughput detection technique of multiplex PCR combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, constructing the characteristic SNP profiles of different strains, and establishing a rapid, accurate and highly sensitive method for the diagnosis of bloodstream infection pathogens.Methods:Seven kinds of pathogens such as common Escherichia coli were selected as target. The multiple PCR reaction conditions was optimized, and the characteristic peaks of each target bacteria were detected by MALDI-TOF MS to establish the joint detect system. Common primer pairs and central homo-sequence primer pairs were designed to analyse the formation of primer dimer. Using simulated bacterial infection blood samples with detection system to determine specificity and sensitivity. One hundred and fifty blood samples from suspected bacteremia patients were collected from June to September 2020 in a hospital in Beijing, and the identification results were compared to traditional identification method of clinical application that are using χ 2 test. Results:The cycle threshold (Ct) value of the central homo-sequence primers that were designed were more than 38, with a delay of 6-10 cycles. The joint mass spectrometry detection system could detect seven kinds of bacteria divided into two groups at the same time. The target bacteria can be detected specific product of the peak, and the clinical strains other than the target strains only had primer peaks. All maps had non-specific miscellaneous peaks. The sensitivity of Escherichia coli could reach 50 CFU/ml, and the detection limit of other bacteria was 100 CFU/ml. The detection results of 150 patients showed that 46 cases were positive by traditional method. The positive rate was 30.67% (46/150), including two cases of mixed infection. Forty-eight cases were positive by mass spectrometry, and the positive rate was 32.0% (48/150), including three cases of mixed infections. The negative coincidence rate was 100% (101/101). The comparison of the two methods showed that the P=0.625>0.01, the Kappa=0.938, the sensitivity and specificity was 97.82%(45/46) and 97.11%(101/104), respectively. There was no significant difference between the two methods, and the results of nucleic acid mass spectrometry could also be used in clinic. Conclusions:The established detection system can not only quickly and accurately detect seven common pathogens causing bloodstream infection, and effectively shorten the time needed for traditional culture and identification, but also can detect multiple bacterial mixed infections at the same time to make up for the possibility of missed detection. Besides, the method can also be used to identify other bacteria.

Objective Toinvestigatestatistically significant peptide peaks as biomarkersto diagnose osteoarticular tuberculosis, matrix-assisted laser desorption/ ionization time of flight mass spectrometry (MALDI-TOF MS) was applied to identify the characteristic fingerprint among the serum of patients with osteoarticular tuberculosis, rheumatoid arthritis and healthy adults.Methods Clinical Study. Serum samples of untreatedpatients with osteoarticular tuberculosis and rheumatoid arthritis were collected from August 2018 to December 2018, and serum samples of healthy adults from physical examination were collected as control. After analysis with MALDI-TOF MS, the serum peptide fingerprint datawas imported into software, and protein polypeptide peaks with obvious differences were screened to establish diagnostic models.Results Established the diagnostic model of osteoarticular tuberculosis and healthy adults with m/z 2943.9, 5929.6, 7615.4 and 9033.8 as differential protein polypeptides, the diagnostic model of osteoarticular tuberculosis and rheumatoid arthritis with m/z 4195.6, 5847.6, 5929.6 and 7748.6 as differential protein polypeptides. To these two models, the sensitivity were 95.00％ and 97.50％, respectively. The specificity were 85.71％ and 88.46％, respectively. The accuracy rates were 89.58％ and 92.39％, respectively. The AUC value of ROC curves were 0.8859 and 0.8709, respectively. Conclusions By mass spectrometry and software analysis, the serum protein polypeptides with statistical difference were found successfully. The related diagnostic modelsarealso established, which has certain reference value for auxiliary diagnosis of osteoarticular tuberculosis.

Anesthesia was done for 36 patients undergoing orthotropic heart transplantation in Bei-jing Anzhen Hospital from April 2015 to November 2016. Anesthesia management for orthotropic heart transplantation and related problems were analyzed and investigated. Anesthesia management protocol for patients with end-stage heart disease was aimed at reducing fluctuation of hemodynamics and avoiding malig-nant arrhythmia. Anesthesia was induced by intravenously injecting diazepam 5-10 mg, etomidate 0. 2-0. 3 mg∕kg or ketamine 1 mg∕kg, sufentanil 1. 0-1. 5 μg∕kg or fentanyl 10-15 μg∕kg and rocuronium 0. 6 mg∕kg. Anesthesia was maintained by continuously infusing dexmedetomidine 0. 3-0. 5μg·kg-1 ·h-1 , ci-satracurium 10 mg∕h and sufentanil 0. 5-1. 0 μg·kg-1 ·h-1 . Pulmonary arterial pressure and donor heart function were monitored using the flow-directed pulmonary artery catheter. Dopamine, epinephrine and iso-prenaline were intravenously infused after cardiopulmonary bypass to maintain circulation stable. Nitroglyc-erin and prostacyclin were intravenously infused to decrease pulmonary arterial pressure. Immunosuppressive therapy was performed with methylprednisone, mycophenolate mofetil and cyclosporine∕FK506. Thirty-two patients were discharged from hospital, and 4 cases died. Among the 4 patients died, 1 patient died of pul-monary hypertension ( pulmonary arterial systolic pressure>67 mmHg) and right heart failure and, 1 patient showed difficulty in weaning from cardiopulmonary bypass and 2 patients died of refractory low cardiac outputand multi-organ failure. Anesthetic management for heart transplantation required an appreciation of the pathophysiological mechanism of heart failure. Invasive monitoring, steady anesthesia induction and mainte-nance, stable hemodynamics in the perioperative period and good donor heart protection were the keys to ensuring anesthesia management for orthotropic heart transplantation.

Objective To investigate the expressions of leukaemia inhibitory factor (LIF) and regulated upon activation,normal T cell expressed and secreted factor (RANTES) in mice with bloodstream infection by 4 different single pathogen and provide research basis for the early diagnosis of bacteriogenous bloodstream infection.Methods CD-1 (ICR,Institute of Cancer Research) mouse models of bloodstream infection with the standard strains of Staphylococcus aureus (S.aureus),Enterococcus faecalis (E.faecalis),Escherichia coli (E.coli) and Klebsiella pneumonia(K,pneumoniae) were established.The serum samples were collected at the 0.5,1,3,6,12,24 and 48 hours after infection and the concentrations of LIF and RANTES in mouse serum of experimental groups and control were detected by Luminex liquid chip system.Results The median lethal dose (LD50) of S.aureus,E.faecalis,E.coli and K.pneumoniae were 8.1 × 108/mL,9.6 × 108/mL,8.1 × 108/mL and 1.1 × 109/mL,respectively.The concentration of serum LIF was significantly increased in 1 hour after infection.The peak concentrations of LIF in the four groups were (51.6±5.0),(73.2±20.8),(7.3 ±0.9)and (6.1 ± 1.2) pg/mL respectively,and the differences were statistically significant compared with the control group (P ＜ 0.01).The concentrations of RANTES in E.faecalis group,E.coli group and K.pneumoniae group were increased after infection for 1 hour and increased significantly after infection for 3 hours.The increased concentrations of RANTES in E.coli group and K.pneumoniae group were more than those in S.aureus group and E.faecalis group.The peak concentrations of RANTES in S.aureus group,E.faecalis group,E.coli group and K.pneumoniae group were (1 929.0-± 25.2),(1 218.1 ± 227.4),(55.7 ± 10.0) and (179.2 ± 9.2)pg/mL,and the differences were statistically significant compared with the control group (P ＜ 0.01).Conclusion The concentrations of LIF and RANTES increased obviously in 1 h after the bacteria entered bloodstream.After 2 days of infections,the levels of LIF and RANTES in E.coli group and K.pneumoniae group were significantly higher than those in S.aureus group and the E.faecalis group.Combined detections of LIF and RANTES may be of certain values to differentiate the infections caused by the pathogens between gram positive and gram negative bacteria.

Objective To reveal the mechanisms of immunological infertility,the method of coimmunoprecipitation(CO-IP) and liquid chromatogram mass/mass (LC-MS/MS) was used to screen sperm membrane proteins which interacting with antisperm antibodies (ASA).Methods This study was designed as a case-control.The disease group including 56 serum samples from 521 cryptogenic infertile patients were screened ASA positive by ELISA and conformed with mixed antiglobulin reaction(MAR).The controls were 31 serum samples which ASA is negative and already possessed healthy offspring.All subjects were enrolled from September 2015 to December 2015 in China PLA General Hospital.Spermatozoa samples from 48donors with normal sperm parameters were from January 2016 to April 2016 in China PLA General Hospital.The purified human sperm membrane proteins were then mixed with serum from disease group (positive for ASA) and control group (not containing ASA).The binding proteins of antisperm antibodies were enriched using CO-IP assay.The immunoprecipitates were separated on sodium dodecyl sulfate-polyactylamide gel (SDS-PAGE),then the binding proteins were cut from the gel and analyzed by LC-MS/MS after the enzymolysis.These proteins could bc idcntified as definition,biological function (s) and subccllular localization with Uniprot database.Results The serum samples from infertile persons (39 females and 17males) were screened ASA positive by ELISA and conformed with MAR.The healthy controls (17 females and 14 males) were ASA-negative in ELISA.Forty proteins that interact with ASA were obtained from the study and these could be divided into three groups:11 antigens detected by control serum samples only,14antigens recognised by both infertile patients and control sera,and 15 antigens specific for patients with ASA.These 15 proteins are Sperm Cation channcl protein 1,Sperm Cation channel protein 3,Sperm Cation channel protein 4,Sperm associated antigen 9,Apolipoprotein A-I,Dynein heavy chain 14,Cylicin-2,Izumo sperm-egg fusion protein 4,Thioredoxin domain-containing protein 2,IQ domain-containing protein H,IQ domain-containing protein F1,Spermatogenesis-associated protein 5,Spermatogenesis-associated protein 5-like protein 1,Sperm acrosome membrane-associated protein 1,E3 ubiquitin-protein ligase RNF 114.Conclusion Fifteen proteins discovered with CO-IP technology and LC-MS/MS analysis could be referred as male immunoinfertility-related antigens and they may hold the great importance in revealing the secret of immunological infertility.

Objective To study the serum peptide fingerprint using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) technology,and find the different peaks with potential significance and establish the diagnosis model of Staphylococcus aureus and Escherichia coli bloodstream infection.Methods To establish ICR mice model of S.aureus and E.coli bloodstream infections,and collect serum samples.The serum samples were purified by weak cation exchange beads,the serum peptide fingerprint was recognized by using MALDI-TOF MS and BioExplorer software between infections group and normal control group.Results Compared with the normal control group,6 peptides were up-regulated,7 peptides downregulated and 8 peptides up-regulated first and then down-regulated in S.aureus infection group;And 5 peptides down-regulated,4 peptides down-regulated first and then up-regulated,and 8 peptides up-regulated first and then down-regulated in E.coli infection group.Conclusion MALDI-TOF MS combined with BioExplorer software may be used as a tool to study the serum peptides of S.aureus and E.coli bloodstream infection,effectively find significant peptides for establishing a diagnosis model of these two bacterial infections,and has a certain value for the diagnosis of bacterial bloodstream infection.

BACKGROUND:Excessive lateral pressure syndrome is often associated with lateral retinacular tension and radiographic patel ar tilt. CT scan displayed that lateral retinacular release can effectively correct patel ar tilt. OBJECTIVE:To study the effect of arthroscopic lateral retinacular release combined with intraosseous dril ing and decomposition in the treatment of excessive lateral pressure syndrome. METHODS:Thirty-two patients with excessive lateral pressure syndrome were treated by arthroscopic lateral release combined with intraosseous dril ing and decomposition. The Lysholm scoring system was used to evaluate the treatment effect. RESULTS AND CONCLUSION:The mean duration of fol ow-up was 12 months. After 1 month, pain of al patients was released or disappeared;after 1 year, pain of 26 cases disappeared basical y. Lysholm scoring system assessment showed 20 cases were rated as excellent, six cases were as good, four cases were as fair and two cases were as poor. The excellent and good rate was 83.6%. The patients’ subjective satisfaction rate was 92.8%. The results indicate that arthroscopic lateral release combined with intraosseous dril ing and decomposition is a good method to treat excessive lateral pressure syndromewere. It has the advantages of less trauma and rapid recovery. Patel ar decomposition has a good effect in the treatment of patel ofemoral pain associated with patel ar tilt outward and lightens articular cartilage degeneration without damage to patel ar cartilage surface.