Objective To evaluate the impact of setting a 4-day incubation time on the detection performance of bloodstream infection(BSI)pathogens by the BacTALERT VIRTUO blood culture system(VIRTUO).Methods Retrospectively collecting blood culture(BC)results from Zhongshan Hospital,Fudan University,from June 2021 to December 2023,and comparing the time-to-detection for clinically significant microorganisms.Results A total of 87 052 BC bottles were collected,of which 7 167(8.23%)were positive.95%of aerobic and anaerobic bottles were reported positive at 65.67 h and 91.2 h respectively.The average(median)time-to-detection for Staphylococcus aureus(S.aureus)was 21.76(16.08)h,Klebsiella pneumoniae(KP)was 15.32(11.65)h,Escherichia coli(E.coli)was 16.14(10.87)h,and Candida species was 35.2(28.5)h.Only 213 bottles(0.24%)were reported positive after 4 days of incubation;among them,88 bottles(41.31%)cultured Propionibacterium acnes(P.acnse).Medical chart reviews for BC reported positive after 96 h(4 days)showed no clinical significance.Simulated growth experiments were conducted on 11 fastidious bacteria,and the median detection time was only 10.8 h,indicating that an incubation time of 4 days is sufficient for fastidious bacteria.Conclusion A 4-day incubation is adequate for VIRTUO system.Reporting 1 day earlier to the doctors can reduce the positive rate of contaminants and the use of antimicrobial agents.

AIM:To investigate the function of mitochondrial reactive oxygen species(mtROS)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)signaling pathway in modulating macrophage polarization in liver fibrosis resulting from iron overload.METHODS:Thirty-two male C57BL/6 mice were randomly allocated into four groups:control group,model group(iron dextran,50 mg/kg),MitoTEMPO(3 mg/kg)group,and MCC950(10 mg/kg)group,comprising eight mice per group.All mice,with the exception of the control group,were administered daily in-traperitoneal injections of iron dextran for a duration of seven consecutive weeks,whereas the control group received equiv-alent volumes of normal saline.Starting in week four,the MitoTEMPO and MCC950 cohorts received their designated treatments through intraperitoneal injection three times weekly.Serum alanine aminotransferase(ALT)and aspartate ami-notransferase(AST)concentrations were assessed through biochemical analysis.Liver tissues were analyzed utilizing HE,Masson,Sirius red and immunohistochemical staining.The concentrations of mtROS were evaluated utilizing the MitoSOX Red probe.Cytokines and polarization markers,such as interleukin-1β(IL-1β),IL-18,IL-6,tumor necrosis factor-α(TNF-α),inducible nitric oxide synthase(iNOS),IL-10,and arginase-1(Arg-1),were quantified via ELISA.Western blot analysis was performed to quantify the protein expression levels of Arg-1,iNOS,NLRP3,apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),and caspase-1.The mRNA expression of NLRP3,ASC,and caspase-1 was assessed using RT-qPCR.Immunofluorescence double labeling was employed to identify M1 and M2 macrophages.RESULTS:(1)In comparison to the control group,the model group demonstrated notable inflammatory cell infiltration,pronounced fibrous tissue hyperplasia,significant disruption of hepatic lobular architecture,and the develop-ment of pseudo-lobules in certain areas.Serum ALT and AST levels were markedly elevated(P<0.01),as were the mRNA and protein expression levels of NLRP3,ASC,caspase-1,and mtROS(P<0.01).Iron overload resulted in markedly ele-vated serum iron,ferritin,total liver iron,and ferrous iron concentrations(P<0.01).Markers indicative of M1 macrophage polarization,including IL-6,TNF-α,and iNOS,exhibited upregulation(P<0.01),whereas M2 markers such as IL-10,Arg-1,and CD206 were significantly downregulated(P<0.01).(2)Compared with model group,inhibiting mtROS or NL-RP3 substantially reduced inflammation and fibrous tissue hyperplasia.ALT and AST levels were markedly diminished(P<0.01),as were the areas of positive staining for α-smooth muscle actin and collagen type Ⅰ(P<0.01).Markers of iron over-load,such as serum iron,ferritin,total liver iron,and ferrous iron,were significantly ameliorated(P<0.01).M1 polariza-tion markers were significantly downregulated(P<0.01).CONCLUSION:The mtROS/NLRP3 signaling pathway facili-tates liver fibrosis caused by iron overload by enhancing macrophage polarization to the M1 phenotype.

Objective The relationship between blood pressure variability(BPV)and short-term neurological prognosis in patients with aneurysmal subarachnoid hemorrhage(aSAH)was investigated.Methods The study conducted a retrospective analysis of clinical data from aSAH patients who underwent surgical treatment and were admitted to the ICU at Beijing Tiantan Hospital,Capital Medical University,from January 2023 to April 2024.BPV was quantitively assessed by calculating the standard deviation(SD),successive variation(SV),coefficient of variation(CV),and range of mean blood pressure(MBP).Patients were divided into two group based on discharge GOS scores:good prognosis[Glasgow Outcome Scale(GOS)4-5]and poor prognosis(GOS 1-3)groups.Comparative analyses were performed to evaluate differences in BPV metrics between the two groups,followed by multivariable logistic regression modeling to adjust for potential confounding factors and elucidate the association between BPV and clinical prognosis.Results A total of 150 patients were included,with 59 in the poor prognosis group and 91 in the good prognosis group.The poor prognosis group exhibited significantly elevated levels of MBP-SD[(9.85±3.20)mmHg vs.(8.04±2.31)mmHg,P<0.001],MBP-SV[(10.37±3.85)mmHg vs.(8.07±2.33)mmHg,P<0.001],MBP-CV(10.00±3.30%vs.8.19±2.33%,P<0.001),and MBP-range[(39.60±13.56)mmHg vs.(32.44±9.78)mmHg,P<0.05]compared to the good prognosis group.Cohen’s d values indicated moderate effect sizes for BPV differences(0.65,0.72,0.63,and 0.61,respectively).Multivariable logistic regression showed that MBP-SD(OR=1.22,95%CI:1.08-1.39,P=0.002)and MBP-SV(OR=1.19,95%CI:1.05-1.35,P=0.007)were independently associated with poor prognosis.Conclusion Elevated MBP-SD and MBP-SV within the first 24 hours postoperative period are independent predictors of unfavorable short-term neurological outcomes in aSAH patients.

OBJECTIVE To evaluate the value of matrix-assisted laser desorption ionization time-of-flight-mass spectrometry(MALDI-TOF MS)in direct identification of carbapenem-resistant Escherichia coli and Klebsiella pneumoniae positive for blood culture.METHODS The blood culture bottles that were positive for E.coli or K.pneumoniae were collected from the patients with bloodstream infection who were treated in Zhongshan Hospi-tal,Fudan University from Jul.2021 to Jun.2023.The isolates were collected by using a gel-contact clotting tube,then the tested strains were respectively mixed with 4 μg/ml of imipenem,4 μg/ml of meropenem and 2 μg/ml of ertapenem for coculture and were incubated at 35 ℃ for 4 and 5 hours,finally,the strains were i-dentified by using the mass spectrum.The minimum inhibitory concentrations(MICs)of the three types of drugs were tested by microbroth dilution method and were set as the golden standards for the test.RESULTS Totally 31 strains of E.coli and 28 strains of K.pneumoniae that were positive in blood culture bottles were collected,both of the effective rates of controlled growth of the E.coli strains were 100.00％after the incuba-tion for 4 and 5 hours,and the effective rates of controlled growth of the K.pneumoniae strains were 9 6.43％and 100.00％after the incubation for 4 and hours,respectively.The sensitivity,specificity,positive predictive value and negative predictive value of the K.pneumoniae strains against the three types of drugs were 100.00％after the incubation for 5 hours.The specificity and positive predictive value of the E.coli strains against imipenem,meropenem and ertapenem were 100.00％after the incubation for 5 hours;the sensitivities were 73.58％,78.93％and 78.93％,respectively;the negative predictive values were 70.60％,75.00％and 75.00％,respectively.CONCLUSIONS MALDI-TOF MS is rapid and accurate for direct identification of the carbapenem-resistant E.coli and K.pneumoniae strains positive in blood culture bottles,and the accuracy reaches at 100.00％for the test of drug-resistant K.pneumoniae strains after the incubation for 5 hours.The method may provide evidence for clinical treatment of bloodstream infections induced by carbapenem-resistant Enterobacteriaceae.

Objective To identify and cluster non-O1/O139 Vibrio cholerae(NOVC)using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS),and evaluate the feasibility of MALDI-TOF MS as a method for the identification and clustering of NOVC.Methods The NOVC was identified by the MALDI-TOF MS equipped with V5 database,V12 database,highly pathogenic bacteria database,and V5 database combined with self-built spectrum projection and analyzed by the principal com-ponent analysis(PCA)and main spectrum projection(MSP)clustering.Results The NOVC was incorrectly identified as Vibrio al-bensis by the MALDI-TOF MS equipped with V5 database or V12 database,while the MALDI-TOF MS equipped with highly pathogenic bacteria database or V5 database combined with self-built spectrum projection could correctly identify NOVC.The PCA clustering of MALDI-TOF MS could distinguish NOVC from other bacterial strains and refine the differentiation of NOVC species to show the dis-tance relationship between NOVC species.Some spectrum projections of NOVC were extremely similar to the reference strains used to establish the database,and MSP clustering could not distinguish the differences between NOVC species.Conclusion The identifica-tion ability of MALDI-TOF MS for NOVC is limited by its database.The MALDI-TOF MS equipped with highly pathogenic bacteria da-tabase or V5 database combined with the self-built spectrum projection can accurately identify NOVC.The PCA clustering of MALDI-TOF MS has certain reference significance for the intra-and inter-species identification and homology analysis of NOVC.

Objective To evaluate the impact of setting a 4-day incubation time on the detection performance of bloodstream infection(BSI)pathogens by the BacTALERT VIRTUO blood culture system(VIRTUO).Methods Retrospectively collecting blood culture(BC)results from Zhongshan Hospital,Fudan University,from June 2021 to December 2023,and comparing the time-to-detection for clinically significant microorganisms.Results A total of 87 052 BC bottles were collected,of which 7 167(8.23%)were positive.95%of aerobic and anaerobic bottles were reported positive at 65.67 h and 91.2 h respectively.The average(median)time-to-detection for Staphylococcus aureus(S.aureus)was 21.76(16.08)h,Klebsiella pneumoniae(KP)was 15.32(11.65)h,Escherichia coli(E.coli)was 16.14(10.87)h,and Candida species was 35.2(28.5)h.Only 213 bottles(0.24%)were reported positive after 4 days of incubation;among them,88 bottles(41.31%)cultured Propionibacterium acnes(P.acnse).Medical chart reviews for BC reported positive after 96 h(4 days)showed no clinical significance.Simulated growth experiments were conducted on 11 fastidious bacteria,and the median detection time was only 10.8 h,indicating that an incubation time of 4 days is sufficient for fastidious bacteria.Conclusion A 4-day incubation is adequate for VIRTUO system.Reporting 1 day earlier to the doctors can reduce the positive rate of contaminants and the use of antimicrobial agents.

Objective:To investigate the clinical distribution characteristics changes in antimicrobial resistance, and carbapenemase resistance genes of Klebsiella pneumoniae isolated from children in Chongqing region during the period of January 2019 to December 2024, providing a basis for the rational use of antibiotics and the prevention and control of nosocomial infections.Methods:An observational study was conducted to retrospectively analyze 5 020 Klebsiella pneumoniae (KP) isolates detected in four hospitals of the Southwest Pediatric Laboratory Specialty Alliance. Antimicrobial susceptibility testing was performed by the minimum inhibitory concentration method combined with the disk diffusion method. Results were interpreted according to the 2024 Clinical and Laboratory Standards Institute (CLSI) standards. Carbapenemase resistance genes were detected by polymerase chain reaction (PCR) combined with Sanger sequencing. WHONET 5.6 was used for resistance analysis and SPSS 19.0 for statistical analysis. The chi-square test was used to assess trends in resistance rates, ESBL detection rates, and resistance rates of different CRKP carbapenemase genotypes from 2019 to 2024. Statistical significance was confirmed if the two-tailed P-value was <0.05. Results:A total of 5 020 strains were isolated, with a detection rate of 5.1% (5 020/99 063). The majority were from sputum (59.2%, 2 970/5 020), followed by pus (17.1%, 857), urine (9.7%, 487), venous blood (6.5%, 326), secretions (2.6%, 130), and other specimens (5.0%, 250).The lowest resistance rate was to amikacin (3.8%), followed by levofloxacin (10.9%), imipenem (19.1%), and meropenem (19.9%). Resistance rates to cefoperazone/sulbactam ( χ2=9.982 0, P=0.001 6), piperacillin/tazobactam ( χ2=10.110 0, P=0.001 5), ceftazidime ( χ2=3.849 0, P=0.049 8), cefotaxime ( χ2=7.605 0, P=0.005 8), cefepime ( χ2=13.510 0, P=0.000 2), aztreonam ( χ2=6.457 0, P=0.011 1), imipenem ( χ2=4.672 0, P=0.030 7), and levofloxacin ( χ2=7.555 0, P=0.006 0) showed an annual increasing trend. The main carbapenemase genes were blaNDM-5 (42.2%, 127/301), blaNDM-1 (33.9%, 102/301), and blaKPC-2 (17.3%, 52/301). Patients with KPC-2-producing strains (median age, 240 days) were older than those with NDM-1/NDM-5-producing strains (median age, 40 days) ( χ2=22.620 0, P<0.000 1). In neonatal wards, the detection rate of NDM-KP was higher than that of KPC-KP (64.6%, 148/229 vs. 26.9%, 14/52, χ2=24.680 0, P<0.000 1), whereas in ICUs, it was lower (6.1%, 14/229 vs. 26.9%, 14/52, χ2=20.450 0, P<0.000 1). Conclusion:In Chongqing region, the isolation rate of K. pneumoniae from sputum was the highest with most cases from neonatal wards. Resistance to carbapenems showed an upward trend. BlaNDM-5 was the predominant genotype in pediatric CRKP. Patients with KPC-KP were older than those with NDM-KP. NDM-KP predominated in neonatal wards, while KPC-KP predominated in ICUs, with KPC-KP showing higher antimicrobial resistance.

Objective To investigate the effect of local vibration therapy(LVT)on fine motor function of upper extremity and surface electromyography characteristics in children with spastic hemiplegic cerebral palsy(SHCP).Methods Sixty children with SHCP were divided into the observation group and the control group using random number table method,with 30 cases in each group.The control group was given conventional rehabilitation treatment(physical therapy,occupational therapy,sensory integration training,physiotherapy and rehabilitation therapy of traditional Chinese medicine),while the observation group received LVT for the affected upper extremity in addition to conventional rehabilitation treatment.Both groups were treated for 12 weeks.Fine motor function of upper extremity on the hemiplegic side[Peabody Developmental Motor Scale-Fine Motor(PDMS-FM),Fine Motor Function Measure(FMFM)]and root mean square(RMS)value of the biceps surface electromyography were compared between the two groups as well as before and after treatment.Results Compared with those before treatment,both groups showed increases in PDMS-FM scores for grasping energy range,visual-motor integration energy range,fine motor development quotient,FMFM scores for upper extremity joint mobility ability,grasping ability,operation ability,hand-eye coordination ability and total scores,but a decrease in RMS value of biceps brachii after treatment(P＜0.05).After treatment,PDMS-FM scores for grasping energy range and visual-motor integration energy range,FMFM scores for upper extremity joint mobility ability and grasping ability and total score were higher in the observation group than those in the control group.The RMS value of biceps brachii was lower in the observation group than that of the control group(P＜0.05).Conclusion LVT can enhance the effect of conventional rehabilitation on fine motor function of upper extremity and electromyography characteristics in children with SHCP.

Objective:To explore the effectiveness of the scaffolding teaching mode from the perspective of deep learning in basic nursing techniques course.Methods:From September 2023 to June 2024, 324 nursing students at Suzhou Vocational Health College in the 2023 cohort were selected as research subjects using convenience sampling. Three classes were designated as experimental group ( n=167), and three classes were designated as control group ( n=157). Control group received the traditional teaching mode, while experimental group received a scaffolding teaching mode from the perspective of deep learning on the basis of control group. Teaching effectiveness was evaluated using course grades, the Academic Self-Efficacy Scale, and the Measure Scale of Autonomous Learning Competencies of Nursing Undergraduates. Results:A total of 324 questionnaires were distributed and 324 valid questionnaires were collected, with a valid response rate of 100.00% (324/324). After teaching, the experimental group's nursing course grades were statistically higher than those of the control group ( P<0.05). After teaching, the experimental group's nursing students demonstrated statistically higher total scores and scores across all dimensions on the Academic Self-Efficacy Scale compared to the control group ( P<0.01). After teaching, the experimental group of nursing students scored statistically higher than the control group on the total score of the Measure Scale of Autonomous Learning Competencies of Nursing Undergraduates, as well as on the dimensions of self-management and learning cooperation ability ( P<0.01) . Conclusions:The implementation of the scaffolding teaching mode from the perspective of deep learning for basic nursing techniques course enhances vocational nursing students' academic performance, academic self-efficacy, and autonomous learning competencies.

Objective To explore and analyze the characteristics and transmission routes of carbapenem-resistant Klebsiella pneumoniae(CRKP)strains in intensive care unit(ICU).Methods From January to October 2023,17 clinical infection isolates(clinical infection group),5 active screening isolates(active screening group),and 7 envi-ronmental isolates(environmental group)of CRKP in the liver surgery ICU of a hospital were selected and analyzed by whole-genome sequencing.The differences in resistance genes,virulence genes,and sequence typing(ST)were compared,and transmission routes were analyzed based on the phylogenetic tree.Results 29 strains of CRKP car-ried 4-18 resistance genes and 52-98 virulence genes,respectively.There were no statistically significant diffe-rences in genotype distribution of resistance genes,the number of virulence genes,and gene types among three groups of CRKP(all P＞0.05).ST showed that 29 CRKP strains mainly consisted of two categories:ST11 and ST15.Based on the phylogenetic tree constructed from the core genome,there were 7 highly homologous groups of CRKP,among which 4 groups had clear epidemiological associations.Conclusion CRKP in ICU carries more re-sistance and virulence genes,and some strains are highly homologous in ST and phylogenetic tree,which may lead to cross transmission.In the future,prevention and control measures should be strengthened to reduce the trans-mission of CRKP.