Objective To perform single-molecule, real-time sequencing of ceftazidime-avibactam (CAZ-AVI)-resistant Pseudomonas aeruginosa and to investigate the mechanism underlying ceftazidime-avibactam resistance in P. aeruginosa. Methods The susceptibility of 89 P. aeruginosa isolates randomly sampled from clinical specimens in Sanming First Hospital Affiliated to Fujian Medical University from November 2021 through July 2023 to common antimicrobial agents was tested, and the minimum inhibitory concentration (MIC) of CAZ-AVI was determined against P. aeruginosa with a broth microdilution assay, with CAZ-AVI MICs of 8 mg/L and lower defined as susceptible and 16 mg/L and higher as resistant. The expression of drug-resistant genes ampC, oxa-488, oprD, mexA, oxa-10, oxa-14, vim and tem was quantified in P. aeruginosa using a real-time quantitative reverse transcription PCR (qPCR) assay. CAZ-AVI-susceptible and -resistant P. aeruginosa isolates from the same case were selected for PacBio single-molecule, real-time sequencing, and sequencing results were subjected to genome structure and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotations. Results The 89 P. aeruginosa isolates showed a relatively high level of resistance to meropenem (75.28%) and imipenem (74.16%) and the highest susceptibility to amikacin (91.01%). There were 49 CAZ-AVI-resistant P. aeruginosa isolates and 40 susceptible isolates. qPCR assay detected lower oprD gene expression in CAZ-AVI-resistant P. aeruginosa isolates [0.104 (2.385)] than in susceptible isolates [0.551 (17.885)] (Z = -2.958, P < 0.01), and there were no significant differences between CAZ-AVI-susceptible and -resistant P. aeruginosa isolates in terms of ampC, oxa-488, mexA or tem gene expression (all P values > 0.05), while oxa-10, oxa-14 and vim gene was expressed in few P. aeruginosa isolates. There were 1 729, 3 936, 3 737 and 3 955 genes in CAZ-AVI-resistant P. aeruginosa isolates PA-762 and PA-M174 and susceptible isolates PA-885 and PA-808 that were annotated to GO terms, with the highest numbers of genes enriched in the molecular function of catalytic activity, high numbers of genes enriched in biological processes of metabolic process, single-organism process and cellular process, and high numbers of genes enriched in cellular components of cell and cell membranes. There were 1 803, 4 084, 3 915 and 4 066 genes in the PA-762, PA-M174, PA-885 and PA-808 isolates enriched in the KEGG signaling pathway, and the majority of genes were enriched in four primary signaling pathways of metabolism, genetic information processing, environmental information processing and cellular process, with the highest number of genes associated with metabolic pathways. Both CAZ-AVI-resistant P. aeruginosa isolates PA-762 and PA-M174 carried multiple efflux pumps systems, including MexAB-OprM, MexCD-OprJ, MexEF-OprN and MexXY-OprM. Single nucleotide substitution was found at position 169 in the DNA sequence of the PA-762 isolate, leading to substitution of serine for glycine at position 57 in the protein sequence, and there are deletions of two bases at positions 307 and 308 in the DNA sequence of the PA-M174 isolate, leading to substitution of threonine for arginine at position 103 in the protein sequence. Conclusion Mutation or downregulation of oprD gene may lead to CAZ-AVI resistance in P. aeruginosa.

Lumbar disc (LD) herniation and aging are prevalent conditions that can result in substantial morbidity. This study aimed to clarify the mechanisms connecting the LD aging and herniation, particularly focusing on cellular senescence and molecular alterations in the nucleus pulposus (NP). We performed a detailed analysis of NP samples from a diverse cohort, including individuals of varying ages and those with diagnosed LD herniation. Our methodology combined histological assessments with single-nucleus RNA sequencing to identify phenotypic and molecular changes related to NP aging and herniation. We discovered that cellular senescence and a decrease in nucleus pulposus progenitor cells (NPPCs) are central to both processes. Additionally, we found an age-related increase in NFAT1 expression that promotes NPPC senescence and contributes to both aging and herniation of LD. This research offers fresh insights into LD aging and its associated pathologies, potentially guiding the development of new therapeutic strategies to target the root causes of LD herniation and aging.
Intervertebral Disc Displacement/metabolism* ; Humans ; Aging/pathology* ; Nucleus Pulposus/pathology* ; Male ; Female ; Transcriptome ; Middle Aged ; Lumbar Vertebrae/pathology* ; Adult ; Cellular Senescence ; Stem Cells/pathology* ; Aged ; Intervertebral Disc Degeneration/metabolism*

OBJECTIVE To explore the risk factors for polymyxin-and carbapenem-resistant Klebsiella pneumoni-ae(PR-CRKP)infection and establish the prediction model.METHODS The clinical data were retrospectively col-lected from the patients with CRKP infection who were treated in the First Affiliated Hospital of Xi'an Jiaotong University from Jan.2023 to Mar.2024.The enrolled patients were divided into the CRKP group and the PR-CRKP group according to the result of drug susceptibility testing for polymyxin B.Totally 203 patients who were treated from Jan.2023 to Dec.2023 were assigned as the modeling group,and 91 patients who were treated from Jan.2024 to Mar.2024 were assigned as the validation group.Multivariate logistic regression analysis was per-formed for the risk factors for PR-CRKP infection,nomogram was built up for prediction of PR-CRKP infection by R software,and the predictive efficacy of the model was evaluated by means of receiver operating characteristic(ROC)curves.RESULTS The result of univariate analysis showed that the proportions of patients who received fi-berobronchoscopy,endotracheal intubation/tracheotomy,were complicated with other carbapenem-resistant or-ganisms(CROs)infections,bloodstream infections,were treated in intensive care unit(ICU)and had the history of exposure to polymyxins and carbapenems 3 months before the admission were higher in the PR-CRKP group than in the CRKP group(P＜0.05);the length of hospital stay and duration of use of polymyxins and carbapene-ms were longer in the PR-CRKP group than in the CRKP group(P＜0.05).Multivariate logistic regression analy-sis indicated that complication with other CROs infections,history of exposure to polymyxins and carbapenems 3 months before the admission,duration of use of polymyxins and ICU stay were the risk factors for the PR-CRKP infection(P＜0.05).The area under the curve(AUC)of the predictive model was 0.898 in the modeling group,with the sensitivity 80.33％,the specificity 84.51％.ROC curve analysis showed that the AUC was 0.901 in the validation group,with the sensitivity 75.00％,the specificity 92.73％.CONCLUSION The prediction model that is established based on the result of multivariate analysis has high value in prediction of PR-CRKP infection.

Objective The aim of this study is to detect the expression of purinergic 2X7 receptor(P2X7R)and the NLRP3 inflammasome in benign prostatic hyperplasia(BPH)tissues and to analyze the clinical correlations.Methods Twelve patients undergoing surgery for BPH were enrolled.Based on the presence or absence of inflammatory cell infiltration in HE-stained tissue sections,the patients were divided into inflammation group and non-inflammation group.Preoperative routine ex-aminations excluded surgical contraindications,and International Prostate Symptom Score(IPSS)questionnaires,urinary flow rate measurements,expressed prostatic secretions(EPS)analysis,and prostate-specific antigen(PSA)tests were conducted.Prostate tissues obtained during surgery were subjected to HE staining,immunofluorescence/Western blot to detect the expression of NLRP3,P2X7R,Caspase-1,Cleaved-Caspase-1,IL-1 β,and TNF-α,and immunofluorescence to assess lymphocyte infiltra-tion.SPSS 26.0 software was used to analyze correlations between the expression levels of NLRP3 and P2X7R in prostate tissues and indicators including Caspase-1,Cleaved-Caspase-1,IL-1 β,TNF-α,lymphocyte count,IPSS score,urinary flow rate,EPS,and PSA.Results In BPH tissues,the expression levels of NLRP3 and P2X7R were positively correlated(P＜0.05).The ex-pression levels of NLRP3 and P2X7R were positively correlated with Caspase-1,Cleaved-Caspase-1,IL-1 β,TNF-α,and lym-phocyte count(P＜0.05).NLRP3 and P2X7R expression levels were positively correlated with white blood cell count in EPS,but showed no correlation with IPSS score,lecithin body count in EPS,maximum urinary flow rate and PSA(P＜0.05).Conclusion P2X7R and NLRP3 in prostate tissues exacerbate local inflammatory responses,which may be an important mecha-nism in BPH development.However,they are not correlated with IPSS score,lecithin body count in EPS,maximum urinary flow rate and PSA.

Microvascular aging is a complex physiological process associated with advancing age.The aging and dysfunction of microvascular endothelial cells serve as critical markers of this phenomenon, characterized by increased expression of inflammatory factors, heightened oxidative stress, and structural damage to the microvasculature.This process is closely linked to age-related diseases such as diabetes in the elderly and cardiovascular disorders, significantly impacting the quality of life and overall health of older adults.Biomarkers, including the reactive hyperemia index, circulating endothelial progenitor cells, and extracellular microvesicles, play a vital role in assessing microvascular health and predicting cardiovascular events.Interventions targeting microvascular aging, which encompass lifestyle modifications, pharmacotherapy, and strategies aimed at preserving microvascular structure and endothelial cell function, hold promise for delaying or reversing the process of microvascular aging.

Objective:To understand the current situation of occupational internal exposure dose research using direct measurement method in China, and to explore the occupational internal dose level of nuclear medicine staff.Methods:From 2023 to April 2024, search on platforms such as China National Knowledge Infrastructure and Chinese Medical Journal Network, to collect research literature on the internal exposure dose monitoring of nuclear medicine staff and discuss the internal exposure dose assessment method. According to the literature reported thyroid 131I activity level at the measuring time, the internal exposure dose level was estimated using the method recommended by GBZ 129-2016. Results:According to existing research reports on direct measurement using portable γ spectrometers, 6.1% of nuclear medicine workers may receive a 131I dose greater than 1 mSv/a. Conclusion:It is necessary to conduct periodic continuous monitoring for personnel whose internal exposure dose may greater than 1 mSv/a, and it is necessary to explore the feasibility of periodic internal exposure monitoring method carried out by hospitals.

Objective:To analyze the values of platelet transforming growth factor-β (TGF-β) and SMAD family member 2 (Smad2) in patients′ peripheral platelets for CRC diagnosis and staging.Methods:Retrospective case-control study. Tumor tissues, paratumor tissues and peripheral blood samples were collected from 248 CRC patients (147 males, 101 females; age 21-93 years) diagnosed in the First Hospital of Jilin University from October 10th, 2020, to March 10th, 2025. Peripheral blood samples were also collected from 40 colorectal adenomatous polyp patients (21 males, 19 females; age 22-74 years) and 75 healthy individuals (43 males, 32 females; age 18-81 years) during the same period. Tissue homogenates and platelets were isolated using tissue disruption and gradient centrifugation, respectively. Total RNA was respectively extracted from tissues and platelets, and the expression levels of TGF-β and Smad2 were quantified by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) expressed as relative quantity 2 -ΔΔCt. Differences of TGF-β and Smad2 expression were compared between CRC tissues and adjacent tissues, as well as among CRC patients, polyp patients, and healthy controls. The relationship of platelet TGF-β and Smad2 expression with pathological features includingtumor stage, pathological type, and metastasis were analyzed. The efficiency of platelet TGF-β, Smad2, and their combination in diagnosing CRC was evaluated using receiver operating characteristic (ROC) curves. Results:The expression levels of TGF-β and Smad2 in CRC tumor tissues[1.09 (0.45, 2.00), 2.93 (0.78, 6.73)] were significantly higher than those in adjacent tissues[0.81 (0.27, 1.50), 1.29 (0.40, 2.63)] ( Z TGF-β=4.54, Z Smad2=6.67, both P<0.001). The expression levels of TGF-β and Smad2 in platelets of CRC patients[2.73(1.53, 4.38), 3.16 (1.58, 4.38)] were significantly higher than those in the colorectal polyp group[1.23(0.70, 2.54), 1.16(0.78, 2.27)] and the healthy control group[0.96(0.51, 1.88), 0.92 (0.55, 1.88)] ( H TGF-β=59.71, H Smad2=78.74, both P<0.001). Platelet TGF-β expression increased progressively with tumor stage (stage 1-4) ( P<0.05), while platelet Smad2 levels were higher in metastatic CRC compared with non-metastatic cases ( P<0.05). ROC curve analysis showed that the area under the curve (AUC) for diagnosing CRC when combining platelet TGF-β and Smad2 was 0.81[95%Confidence interval( CI) 0.77—0.86], which was 0.90 (95% CI 0.86—0.93) if adding serum carcinoembryonic antigen (CEA). Conclusion:Platelet TGF-β and Smad2 expression correlates with the diagnosis and staging of CRC, demonstrating potential as liquid biopsy biomarkers for colorectal malignancies.

Objective:To detect the biological markers related to Alzheimer's disease(AD)in the peripheral blood of AD patients,and to explore the activities and levels of the antioxidant function indexes and the expressions of related genes and proteins in the blood of AD patients and the changes after intervention of selenium-rich food.Methods:The Mini-Mental State Examination(MMSE)combined with electroencephalogram or brain CT and clinician diagnosis were used for screening AD.Fifty-six elderly patients with AD aged 75-90 years old were selected.Among them,28 cases were selected as normal diet group for AD(AD group),and 28 cases were selected as dietary selenium intervention group(Se-AD group).The patients in Se-AD group were given daily dietary selenium supplementation(increaseing dietary selenium by 15-20 μg per day)for 3 months.Meanwhile,30 people with the same age were selected as healthy control group.The activities of serum superoxide dismutase(SOD),cholinesterase(CHE),and glutathione peroxidase(GSH-Px)and the levels of serum malondialdehyde(MDA),homocysteine(Hcy),and nitric oxide(NO)as well as reagent kit the levels of serum β-amyloid protein(Aβ),and microtubule-associated protein(Tau)and phosphorylated microtubule-associated protein(p-Tau)of the subjects in various groups were detected by and enzyme-linked immunosorbent assay(ELISA)method;the expression levels of apolipoprotein E4(ApoE4),presenilin 1(PS1),presenilin 2(PS2),cysteinyl aspartate specific proteinase 3(Caspase3),sorting associated protein receptor 1(SORL1),β-site amyloid precursor protein cleaving enzyme 1(BACE1),hypoxia-inducible factor 1(HIF1),nuclear factor-kappa B(NF-κB),β-amyloid precursor protein(APP),protein kinase C(PKC),and Aβ mRNA in peripheral blood of the subjects various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method.Results:Compared with healthy control group,the serum SOD activities of the patients in Se-AD group and AD group were significantly decreased(P＜0.05),while serum CHE activity and the levels of MDA and Hcy were significantly increased(P＜0.05);the serum GSH-Px activity of the patients in AD group was significantly decreased(P＜0.05),and the level of NO was significantly increased(P＜0.05).Compared with Se-AD group,serum CHE activity and the level of Hcy of the patients in AD group were significantly increased(P＜0.05).The expression levels of ApoE4,PS1,Caspase3,BACE1,NF-κB and APP mRNA of the patients in Se-AD group and AD group were significantly increased(P＜0.05),and the expression levels of PKC mRNA were significantly decreased(P＜0.05);the expression level of PS2 mRNA of the patients in AD group was significantly increased(P＜0.05),and the expression levels of Aβ mRNA of the patients in Se-AD group and AD group were significantly increased(P＜0.05).Conclusion:The activities of serum SOD,GSH-Px and CHE and the levels of MDA,Hcy and NO,the levels of Aβ,Tau and p-Tau proteins,and the expression levels of ApoE4,PS1,Caspase3,BACE1,NF-κB,PKC,PS2,Aβ and APP mRNA in peripheral blood of the AD patients may vary and can be used for clinical diagnosis of the AD patients.Selenium-rich food can improve AD to some extent,and its mechanism is related to reducing the oxidative damage of brain tissue and decreasing the expression of AD related genes PS2 and Aβ.