Objective To investigate the risk factors for mortality and bleeding complications in extracorporeal membrane oxygenation(ECMO)treated patients and to evaluate the impact of thrombocytopenia severity on the prognosis of ECMO therapy.Methods A total of 153 patients who received ECMO treatment at Peking University Third Hospital between January 2013 and September 2024 were en-rolled in this study.The patients were divided into death group(n=97)and recovery group(n=56)based on their final outcomes.Additionally,the patients were categorized into bleeding group(n=104)and non-bleeding group(n=49)based on the occurrence of bleeding complications during ECMO.Clinical baseline characteristics and extreme laboratory values during ECMO were compared be-tween groups.Logistic regression was used to analyze the risk factors for mortality and bleeding.The patients were further divided,based on the initial platelet(PLT)values on the day of catheter placement and the lowest platelet count during ECMO,into normal group(PLT≥ 100× 109/L),moderate reduction group[PLT=(50～99)× 109/L],and severe reduction group(PLT＜50× 109/L).Kaplan-Meier analysis was used to compare survival rates among these groups.The patients in the moderate and severe reduction groups were further divided into a platelet transfusion group and a non-transfusion group,and the outcomes and complication rates were com-pared.Results The recovery group had a higher proportion of myocarditis,higher minimum values of PLT,Hb,and Fib,and higher initial PLT values,while the maximum values of lactic dehydrogenase(LDH),total bilirubin(T-Bil),prothrombin time(PT),and procalcitonin(PCT)were lower(all P＜0.05)with significant differences.Logistic regression showed that age and maximum PCT were independent risk factors for mortality(OR=1.025 and 1.015 respectively,all P＜0.05).The bleeding group had longer ECMO dura-tions,more plasma transfusions,lower minimum Hb values,and higher maximum values of WBC,neutrophils(Neu),and APTT(all P＜0.05)with statistical differences.The minimum PLT value,maximum WBC value,and maximum APTT value were independent risk factors for bleeding complications(OR=0.986,1.062,and 1.004 respectively,all P＜0.05).Kaplan-Meier analysis showed that the patients in the severe reduction group had lower survival rates,regardless of whether the grouping was based on initial or minimum platelet counts(all P＜0.05).Platelet transfusion improved the mortality in the severe reduction group(P＜0.05)but had no effect on the moderate reduction group.Conclusion Age and peak value of PCT are the risk factors for mortality in ECMO patients,while mini-mum PLT count,peak value of WBC and APTT are the risk factors for bleeding complications.Early intervention for infection and in-flammation during ECMO may improve the outcome of patients.Severe thrombocytopenia during ECMO therapy increased the risk of mortality,and targeted platelet transfusion may improve the survival of these patients.

Objective To prepare in-house coagulation factor Ⅷ(F Ⅷ)inhibitor-positive control material and evaluate its perform-ance.Methods Frozen plasma samples from hemophilia A patients with positive factor Ⅷ inhibitors were pooled,and diluted with Owren's Veronal Buffer(OVB)to 1 BU/mL of the inhibitor concentration in the mixture,then aliquoted and freeze-stored.The homo-geneity and stability of the in-house quality control material were verified,and its suitability was further assessed through intra-laborato-ry reproducibility among different technologists and inter-laboratory comparisons.Results Twenty-one aliquots were randomly tested for homogeneity assessment,yielding an average of 1.05 BU/mL(range 0.9-1.15 BU/mL),with a standard deviation(SD)of 0.083 and coefficient of variation(CV)of 7.90％.The freshly prepared inhibitor-positive control samples contained a concentration of 1.03 BU/mL.After storage at-80℃ for 24 hours,1 week,1 month,2 months,3 months,4 months,5 months,6 months,7 months,8 months,and 9 months,thawed the samples showed relative deviations of 9％,0％,10％,9％,14％,15％,6％,0％,-10％,-5％,and 2％,respectively.The intra-laboratory CV value from different technologists at this center was 7.28％,and the inter-labora-tory CV across different centers was 18.75％.Conclusion The prepared in-house positive control material of Factor Ⅷ inhibitor ex-hibited adequate uniformity and stability.

Objective To analyze the clinical value of fungus(1,3)-β-D glucan test magnetic particle chemiluminescence immunoas-say(G-CLIA)for diagnosis of invasive fungal disease(IFD).Methods A total of 509 patients with clinically suspected IFD in Qilu Hospital of Shandong University from 1 March to 30 April,2023 were collected.According to the inclusion criteria,the 509 patients were grouped into IFD group(141 patients)and non-IFD group(368 patients).The sensitivity,specificity,accuracy,positive predic-tive value and negative predictive value of G-CLIA were analyzed,and the consistency of the results of G-CLIA with G test colorimetric method,fungal smear microscopy or culture and metagenomics next-generation sequencing(mNGS)was comparatively analyzed.Re-sults The sensitivity and specificity of G-CLIA were 88.65％and 96.47％,respectively,and the positive percentage agreement of G-CLIA with G test colorimetric assay,fungal smear microscopy or culture,and mNGS were 92.19％,75.86％,and 75.00％,respective-ly,and the consistency of G-CLIA with G test colorimetric assay was the highest(kappa value≥ 0.75).Conclusion G-CLIA has high sensitivity and specificity for detecting IFD with excellent diagnostic value.Combined with the fully automated chemiluminescence analy-zer,G-CLIA test is fast and has a high throughput,which provides a new option for the clinical diagnosis of IFD.

Objective To analyze the drug resistance characteristics,molecular typing,and biological properties of carbapenem-resist-ant Klebsiella pneumoniae(CRKP).Methods A retrospective analysis was conducted on 31 non-repetitive CRKP strains collected clinically from April 2019 to May 2021 at the Third People's Hospital of Nantong.The Vitek 2 Compact microbial analysis system was used for bacterial identification and in vitro drug susceptibility analysis.The broth dilution method was used to determine the minimum inhibitory concentration(MIC)of polymyxin B.The disk diffusion testing was performed to supplement the susceptibility of five com-monly used antibiotics:ertapenem,cefotaxime,cefoxitin,cefoperazone-sulbactam,and tigecycline.The carbapenemase-resistance phenotype of CRKP strains was initially determined by a combined assay of modified carbapenem inactivation method(mCIM)and ED-TA-carbapenem inactivation method(eCIM).Certain carbapenemase resistance genes(blaKPC,blaNDM,blaIMP,blaVIM,and blaOXA-48),AmpC enzyme genes(blaDHA,bla ACC,blaCIT,blaEBC,blaMOX,and blaFOX),extended-spectrum β-lactamases(ESBLs)genes(blaSHv,blaTEM,and blaCTX-M),and nine virulence genes were amplified by PCR and subsequently verified by sequencing.The stringing test was used to screen for hypermucoviscous phenotype strains.The growth curves in vitro and biofilm formation assays,and multilocus se-quence typing(MLST)were performed on 31 isolates.Outer-membrane proteins were extracted and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)to evaluate the expressions of OmpK35 and OmpK36.Results All the 31 isolates were resistant to ampicillin/sulbactam,ampicillin,aztreonam,cefazolin,ceftriaxone,cefotaxime,cefuroxime,ciprofloxacin,pip-eracillin,piperacillin/tazobactam,meropenem,and ertapenem with resistance rate of 100％.The resistance to polymyxin B was ob-served in 32.26％,whereas tigecycline retained 100％susceptibility.In terms of MLST,three sequence types(STs)were identified,with ST15 being the most prevalent,accounting for 61.29％(19/31)of the isolates.All strains produced serine carbapenemase,and only blaKPC-2 was detected among carbapenem resistance genes.The virulence genes fimH and entB were present in all strains(100％,31/31),while the detection rate of mrkD was 80.64％(25/31).Some strains carried virulence genes such as rmpA,rmpA2,and other virulence genes,whereas magA gene was not detected in any isolate.The carriage rates of rmpA2,iutA,and mrkD were higher in ST11 strains than in ST15 strains.The string test was positive in 38.71％of the strains.The growth test showed that there was no significant difference observed in the growth curves among all strains in vitro,and all were able to form biofilms with varying degrees.All ST11 strains exhibited OmpK36 protein alterations,while OmpK35 protein was intact in the 31 strains.Conclusion CRKP strains in this hospital showed high drug-resistance rate,and ST15 was the predominant sequence type.All the isolates carried blaKPC-2 and virulence genes.Enhanced molecular surveillance and strengthened prevention and control measures of CRKP infection are urgently needed.

Objective To establish a flow cytometry method for detecting C1s protein on platelet surface and preliminarily explore its potential application value in the auxiliary diagnosis of primary immune thrombocytopenia(ITP).Methods C1s-conjugated 2 μm car-boxylated magnetic beads(C1s beads)were prepared and used as quality control particles.Fluorescein isothiocyanate(FITC)-labeled anti-C1s antibody was employed as the detection antibody to develop a flow cytometric assay for detecting C1s protein expression on platelets.The intra-assay and inter-assay precision,as well as the dilution linearity of the method,were evaluated.Subsequently,the expression levels of C1 s protein on the surface of platelets were compared among the ITP group,the non-ITP thrombocytopenia group,and the healthy control group.Results Light microscopy showed that both unconjugated carboxylated magnetic beads(blank beads)and C1s-conjugated beads were uniformly dispersed without aggregation.Under fluorescence microscopy,C1s beads exhibited strong yellow-green fluorescence,whereas the blank beads showed no fluorescence signal.The established flow cytometry assay exhibited ac-ceptable precision,with intra-assay coefficient of variation(CV)values of 7.02％,7.12％,and 3.91％for low,medium,and high con-centrations of C1s beads,respectively,and inter-assay CV values of 13.49％,6.15％,and 0.78％,respectively.The dilution linearity was satisfactory,coefficient of determination(R2)=0.998 8.Clinical sample testing revealed that the proportion of C1s-positive plate-lets in ITP group(2.56±0.79)％was significantly higher than that in healthy control group(0.23±0.18)％and the non-ITP thrombo-cytopenia control group(0.22±0.10)％,with statistically significant differences(both P＜0.05).Conclusion This study successfully established a stable and reliable flow-cytometry method for quantifying C1s expression on platelet surface and preliminarily demonstrated that C1s expression is significantly elevated on platelets of ITP patients,suggesting that C1s could serve as a potential auxiliary diag-nostic marker for ITP.

Objective To establish autoverification rules for six routine coagulation assays(PT,APTT,TT,Fib,DD,and FDP)based on the stratification of outpatients and inpatients,in accordance with CLSI AUTO-10A,AUTO-15,and WS/T 616-2018 guide-lines,and to validate the feasibility of this stratified strategy with clinical data while optimizing verification efficiency.Methods A to-tal of 323 451 coagulation test results from Zhongshan Hospital,Fudan University in 2022 were retrospectively analyzed to define auto-verification rules involving critical values,instrument flags,logical rules,historical comparison,and numerical ranges.A stratified au-toverification system was established by applying distinct rules for outpatient and inpatient populations.Subsequently,the rules were op-timized using 87 830 coagulation test results from January to March 2024,and the consistency between autoverification and manual veri-fication was prospectively evaluated using 33 968 consecutive coagulation specimens collected in April 2024.Results A stratified au-toverification system was successfully developed,comprising a total of 53 rules.The pass rate of overall verification was 77.16％(26 210/33 968),with a true-positive rate of 19.64％(6 672/33 968),a false-positive rate of 3.20％(1 086/33 968),a true-nega-tive rate of 77.16％(26 210/33 968),and no false negatives were detected.Conclusion The proposed autoverification system signifi-cantly improved verification efficiency.The stratified design based on outpatient and inpatient populations effectively minimized the risk of false negatives,and may provide a novel approach for the further development and optimization of coagulation test autoverification.

Objective To establish and validate an autoverification system for pediatric coagulation tests and apply it in clinical prac-tice.Methods A total of 31 633 specimens collected at the Children's Hospital of Soochow University,from August 1,2023 to De-cember 31,2023,were used to establish coagulation auto-verification rules,which were subsequently validated with 6 704 specimens collected during January 1-31,2024.The samples were analyzed using the SYSMEX fully automatic blood coagulation analyzer assem-bly line.Auto-verification rules were established based on quality control status,instrument alerts,sample trait,numerical anomalies,logical rules,linear deviations,specimen integrity,and delta checks.The pass rate and consistency rate following the auto-verification were subsequently evaluated.Results A total of 37 auto-verification rules were established,covering abnormal results for PT-INR,APTT,Fib,TT,DD,FDP and AT together with aberrant reaction curves,specimen status and instrument alerts.The pass rate of auto-verification in the establishment group was 57.44％,and the concordance rate between auto-verification and manual review was 99.82％,while the pass rate of auto-verification was 59.96％in the validation set,and the concordance rate between auto-verification and manual review was 100％.Conclusion A set of auto-verification rules for pediatric coagulation reports was established and imple-mented in routine practice,significantly expediting the efficiency of report review and effectively shortening the turn-around time(TAT).

Objective To investigate the prognostic value of tissue plasminogen activator-inhibitor complex(tPAIC)for mortality in patients with sepsis associated thrombocytopenia(SAT).Methods A retrospective analysis was conducted on the clinical data of 74 patients with SAT admitted to the Intensive Care Unit of the 908th Hospital of the Joint Logistics Support Force of PLA between Septem-ber 2017 and December 2021.Based on 28-day follow-up for outcomes,the patients were divided into survival group(43 cases)and death group(31 cases).The Boruta algorithm based on random forest and Lasso regression algorithm were used to screen the risk fac-tors from a total of 29 candidate variables,including age,sex,SOFA score on admission,APACHE Ⅱ score,and various hematological indexes,(e.g.,white blood cell count).The time-dependent receiver operating characteristic(ROC)curves and the survival analysis were performed for the key variables identified.Results Compared with the survival group,the nonsurvivors in death group had signif-icantly higher SOFA and APACHE Ⅱ scores.The levels of thrombomodulin(TM),thrombin-antithrombin complex(TAT),and tissue plasminogen activator inhibitor complex(tPAIC)were also markedly elevated in death group(P＜0.05).Similarly,PT,APTT,TT,FDP,and D-dimer in the death group were significantly higher than those in survivalroup,whereas the fibrinogen levels were signifi-cantly decreased(P＜0.05).The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(T-Bil),and lactate(Lac)levels were likewise significantly elevated in the death group.Random forest and Lasso regression analysis identified SOFA score,tPAIC,INR,and Lac as the independent risk factors of mortality in the patients with sepsis-associated thrombo-cytopenia(SAT)patients.Time-dependent ROC curve analysis showed that a higher C-index and average area under the curve(AUC)in tPAIC than any other variables for predicting the prognosis of SAT patients.Kaplan-Meier survival analysis stratified by the optimal cutoff value of tPAIC(38.9 ng/mL)revealed that a 2.31-fold higher 28-day survival rate in the patients with tPAIC＜38.9 ng/mL group compared those with tPAIC ≥38.9 ng/mL.Conclusion tPAIC demonstrates significant predictive value for mortality in SAT patients.

Objective To investigate the mutation spectrum of F7 gene and its clinical implications in patients with coagulation factorⅦ(FⅦ)deficiency in the southeastern Chinese population,and to analyze the correlations among genotype,FⅦ activity(FⅦ:C),and bleeding risk.Methods A retrospective analysis was conducted on 66 probands diagnosed with FⅦ deficiency between 2010 and 2024 at The First Affiliated Hospital of Wenzhou Medical University.The clinical data,bleeding scores according to ISTH(Internation-al Society on Thrombosis and Haemostasis),the results of coagulation function tests,and F7 gene sequencing data were collected and analyzed.Results Among the 66 probands,59 cases exhibited severe FⅦ deficiency,of whom 37 presented bleeding symptoms,pri-marily gingival bleeding,epistaxis,and menorrhagia.The most frequent mutation:sp.His408Gln,p.Cys10Profs * 16,and p.Cys389Gly,were clustered in exon 8.Prothrombin time(PT)showed a significant positive correlation with ISTH bleeding scores(P＜0.05),while FⅦ:C demonstrated weak predictive power for bleeding risk.Conclusion Exon 8 and the S1 peptide region of the F7 gene were identified as mutation hotspots,and PT was highlighted as an effective tool for evaluating bleeding risk.Although FⅦ:C levels exhibited only a limited correlation with bleeding risk,genetic mutation analysis provided crucial insights for the molecular diag-nosis and clinical management of FⅦ deficiency.

Objective To explore the risk factors of acute respiratory failure(ARF)in adult patients with community-acquired pneu-monia(CAP),and thereby construct and validate the efficacy of nomogram model.Methods The clinical and laboratory data of 172 adult CAP patients admitted to Taikang Xianlin Drum Tower Hospital affiliated to Nanjing University School of Medicine from January 2018 to December 2021 were retrospectively collected.The patients were divided into two groups based on whether they had concurrent ARF.After the comparison for the differences of single factor between the two groups,collinearity analysis was assessed.The risk fac-tors were then screened by binary logistic regression analysis with forward stepwise regression method.A nomogram model was subse-quently constructed and the discrimination and accuracy of the model were evaluated by ROC and colibration curves.Results Among the 172 CAP patients,53 cases(30.8％)developed ARF.The results of univariate analysis showed that the CAP patients with concur-rent ARF group had higher age,CURB-65 score and inflammatory markers than the non-concurrent ARF group,and the incidence of complex infection(culturing two or more pathogenic bacteria)was high.The values of CRP(C-reactive protein)and BUN/Alb(blood urea nitrogen/albumin)were significantly different between the two groups(53.910[25.900,101.200]vs.23.300[6.800,48.930],0.231[0.160,0.302]vs.0.123[0.089,0.171],P＜0.05).Multivariate analysis indicated:glucose(Glu)≥6.06 mmol/L(odds ra-tio(OR):2.737,95％confidence interval(CI):1.116-7.037),AST(aspartate aminotransferase)≥22.5 U/L(OR:4.291,95％CI:1.779-11.120),fibrinogen(Fib)≤3.83 g/L(OR:3.955,95％CI:1.631-10.237),uric acid(UA)188.07 μmol/L(OR:4.617,95％CI:1.859-12.489),BUN/Alb≥0.15 mmol/g(OR:6.381,95％CI:2.423-18.513),total number of multicomor-bidity≥3(OR:6.191,95％CI:2.088-21.905)were the risk factors(P＜0.05).All the screened indicators were incorporate into the nomogram model and its efficacy was verified.The results showed that the area under the curve of the model was 0.888[95％CI:0.840-0.935](P＜0.05),the sensitivity was 0.868,and the specificity was 0.790.The calibration curve showed that the predicted probability of adult CAP patients-associated with ARF was in good consistency with the observed probability(Briser Score:0.125;H-L test:x2=7.563,P=0.477).Conclusion The established model has a good ability to predict adult CAP associated with ARF,and can provide a reference basis for early clinical prediction and intervention treatment.