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.

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.

(±)-Talapyrones A-F (1-6), six pairs of dimeric polyketide enantiomers featuring unusual 6/6/6 and 6/6/6/5 ring systems, were isolated from the fungus Talaromyces adpressus. Their structures were determined by spectroscopic analysis and HR-ESI-MS data, and their absolute configurations were elucidated using a modified Mosher's method and electronic circular dichroism (ECD) calculations. (±)-Talapyrones A-F (1-6) possess a 6/6/6 tricyclic skeleton, presumably formed through a Michael addition reaction between one molecule of α-pyrone derivative and one molecule of C₈ poly-β-keto chain. In addition, compounds 2/3 and 4/5 are two pairs of C-18 epimers, respectively. Putative biosynthetic pathways of 1-6 were discussed.
Polyketides/isolation & purification* ; Talaromyces/chemistry* ; Stereoisomerism ; Molecular Structure ; Circular Dichroism ; Pyrones/chemistry*

Objective:To identify the relevant factors for the first-course remission of acute myeloid leukemia (AML) and to develop a predictive model as well as assess its predictive capability.Methods:Clinical data of 749 patients newly diagnosed with AML admitted to the Department of Hematology, the First Affiliated Hospital, Zhejiang University, School of Medicine from January 1, 2019, to April 30, 2023, were collected and randomly divided into training and validation sets. Multivariate logistic regression analysis was conducted to determine variables associated with complete remission in the first course of induction therapy, and a predictive model was established based on these variables. The receiver operating characteristic (ROC) curve of the predictive model was plotted, and the area under the curve (AUC) was calculated.Results:The indicators predicting the first remission course included peripheral blood white blood cell count during onset, CBF::MYH11 fusion gene, CEBPA bZIP region mutation, myelodysplastic syndrome-related gene mutation, and induction chemotherapy regimen selection as independent factors for the first remission course. The model’s area under the training and validation curves was 0.738 (95% CI: 0.696-0.780) and 0.726 (95% CI: 0.650-0.801), respectively. The Hosmer-Lemeshow test results yielded P-values of 0.993 and 0.335, respectively. Conclusion:In this study, the developed model demonstrates a strong predictive capability for the efficacy of the first course of patients with AML, providing valuable guidance to clinicians in assessing patient prognosis and selecting appropriate treatment strategies.

Objective:To explore the molecular epidemiological characteristics of human metapneumovirus (HMPV) in children with acute respiratory infection (ARI) in Beijing from 2023 to 2024.Methods:In the longitudinal study, 9 834 children with ARI were enrolled from August 2023 to December 2024, including the influenza-like illness (ILI) group from emergency and outpatient department receiving influenza virus (Flu) and HMPV test and the ARI inpatient group for 13 common respiratory pathogen screening test including HMPV, Flu, respiratory syncytial virus, and so on. All respiratory samples positive with HMPV were genotyped by amplifying and sequencing of G gene and further phylogenetic analysis. The χ2 test and Wilcoxon rank-sum test were used to compare the positive rate and basic clinical data of the 2 groups. Results:Among 9 834 enrolled patient, there were 5 276 male and 4 558 female children, with age 5.4 (1.9, 8.2) years. In ILI group of 1 460 patients, there were 83 cases (5.7%) positive for HMPV, with the age 4.9 (3.6, 6.6) years and children under 6.0 years old 59 cases (71.1%). Among 8 374 ARI inpatients, there were 256 cases (3.1%) positive for HMPV, with age 3.5 (1.3, 6.4) years and children under 6.0 years old 188 cases (73.4%). The HMPV positive rate and the age of children positive for HMPV in ARI inpatient group were significantly lower than that in ILI group (both P<0.001). In December, 2024, the HMPV positive rates of ILI and ARI inpatient group (21.3% (17/80), 15.0% (47/314)) were significantly higher than the total positive rates of each group (both P<0.001). Among 279 subtyped specimens, there were 155 cases (55.6%) belonging to genotype A and 124 cases (44.4%) belonging to genotype B. Sub-lineage A2.2.2 containing 111nt-insertions was predominate one in 2023 with positive ratio 89.2% (91/102), and B2 was predominate in 2024 with positive ratio 64.4% (114/177). Conclusions:From 2023 to 2024, the positive rate of HMPV in the ILI group was higher than that in the ARI inpatient group, suggesting a common epidemic of HMPV infection. Children positive for HMPV in the ARI inpatient group were younger than that in the ILI group. A severe epidemic of HMPV was observed in the winter of 2024, which requires attention. Sub-lineage A2.2.2 with 111nt-duplicate insertions and B2 were the predominant epidemic strains in 2023 and 2024, respectively.

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 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.