Rapid and ultrasensitive detection of pathogen-associated biomarkers is vital for the early diagnosis and therapy of bacterial infections.Herein,we developed a close-packed and ordered Au@AgPt array coupled with a cascade triggering strategy for surface-enhanced Raman scattering(SERS)and colorimetric identification of the Staphylococcus aureus biomarker micrococcal nuclease(MNase)in serum samples.The trimetallic Au@AgPt nanozymes can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine(TMB)molecules to SERS-enhanced oxidized TMB(oxTMB),accompanied by the color change from colorless to blue.In the presence of S.aureus,the secreted MNase preferentially cut the nucleobase AT-rich regions of DNA sequences on magnetic beads(MBs)to release alkaline phosphatase(ALP),which subsequently mediated the oxTMB reduction for inducing the colorimetric/SERS signal fade away.Using this"on-to-off"triggering strategy,the target S.aureus can be recorded in a wide linear range with a limit of detection of 38 CFU/mL in the colorimetric mode and 6 CFU/mL in the SERS mode.Meanwhile,the MNase-mediated strategy characterized by high specificity and sensitivity successfully discriminated between patients with sepsis(n=7)and healthy participants(n=3),as well as monitored the prog-nostic progression of the disease(n=2).Overall,benefiting from highly active and dense"hot spot"substrate,MNase-mediated cascade response strategy,and colorimetric/SERS dual-signal output,this methodology will offer a promising avenue for the early diagnosis of S.aureus infection.

Rapid and ultrasensitive detection of pathogen-associated biomarkers is vital for the early diagnosis and therapy of bacterial infections. Herein, we developed a close-packed and ordered Au@AgPt array coupled with a cascade triggering strategy for surface-enhanced Raman scattering (SERS) and colorimetric identification of the Staphylococcus aureus biomarker micrococcal nuclease (MNase) in serum samples. The trimetallic Au@AgPt nanozymes can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) molecules to SERS-enhanced oxidized TMB (oxTMB), accompanied by the color change from colorless to blue. In the presence of S. aureus, the secreted MNase preferentially cut the nucleobase AT-rich regions of DNA sequences on magnetic beads (MBs) to release alkaline phosphatase (ALP), which subsequently mediated the oxTMB reduction for inducing the colorimetric/SERS signal fade away. Using this "on-to-off" triggering strategy, the target S. aureus can be recorded in a wide linear range with a limit of detection of 38 CFU/mL in the colorimetric mode and 6 CFU/mL in the SERS mode. Meanwhile, the MNase-mediated strategy characterized by high specificity and sensitivity successfully discriminated between patients with sepsis (n = 7) and healthy participants (n = 3), as well as monitored the prognostic progression of the disease (n = 2). Overall, benefiting from highly active and dense "hot spot" substrate, MNase-mediated cascade response strategy, and colorimetric/SERS dual-signal output, this methodology will offer a promising avenue for the early diagnosis of S. aureus infection.

Objective:To establish a method for detecting serum non-ceruloplasmin-bound copper (NCC) by immunoprecipitation-inductively coupled plasma mass spectrometry (IP-ICP-MS) and evaluate its clinical application.Methods:Methodological evaluation research. Immunoprecipitation was first used to separate serum ceruloplasmin, followed by detection of serum NCC levels using ICP-MS. Two levels of quality control serum were reconstituted to determine the limit of detection (LOD), limit of quantitation (LOQ), accuracy, and precision of the self-developed method. The serum samples of 131 healthy individuals (healthy group) and 69 first-time diagnosed Wilson′s disease (WD) patients(WD group) from November 2023 to June 2024 in the Affiliated Hospital of Institute of Neurology, Anhui University of Traditional Chinese Medicine were collected. Using self-developed method detected the serum NCC levels, established the healthy reference intervals, and NCC levels between the two groups were compared using the Mann-Whitney U test. Results:The calibration curve exhibited excellent lineary across the concentration range of 1.562 to 300.000 μg/L, as demonstrated by coefficient of determination ( R2) of 0.999. The self-developed NCC method exhibited that the LOD was 0.99 μg/L, the LOQ was 3.29 μg/L, the accuracy (spike and recovery experience) was 87.67%?106.27%, and the intra-batch and inter-batch imprecision expressed as coefficient of variation ( CV) was 2.8%?7.3%, and the healthy reference range was 34.31-71.79 μg/L. The serum NCC levels in the WD group were 102.39 (74.38, 144.04) μg/L, which was significantly higher than those in healthy group (51.45±10.34) μg/L ( Z=?7.967, P<0.01). Conclusions:The established IP-ICP-MS method for detecting serum NCC meets the required analytical performance criteria. It is simply operative, highly sensitive, and provides accurate and reliable results, which could be used for clinical detection.

Objective To construct an MRI radiomics model of cerebral damage following carbon monoxide(CO)poisoning.Methods Ninety patients with CO poisoning and 70 healthy controls(HC)were scanned by structural MRI.All patients were randomly divided into training set(n=128)and test set in a ratio of 8∶2.The radiomics features of 9 regions of interest(ROI)were automatically extracted,and each ROI was subjected to 5-fold cross-validation and 9 different types of classifiers to construct a diagnostic model.Receiver operating characteristic(ROC)curve and decision curve analysis(DCA)were used to evaluate the diagnostic efficacy and clinical value of the model.Results The model based on the radiomics features of the whole brain gray matter(GM)and the left globus pallidus(LGP)had better diagnostic efficacy and clinical application value.The area under the curve(AUC)in the training set and the test set were 0.813,0.815 and 0.806,0.806,respectively.Conclusion The GM-based radiomics model can effectively diagnose cerebral damage following CO poisoning.

Objective To construct an MRI radiomics model of cerebral damage following carbon monoxide(CO)poisoning.Methods Ninety patients with CO poisoning and 70 healthy controls(HC)were scanned by structural MRI.All patients were randomly divided into training set(n=128)and test set in a ratio of 8∶2.The radiomics features of 9 regions of interest(ROI)were automatically extracted,and each ROI was subjected to 5-fold cross-validation and 9 different types of classifiers to construct a diagnostic model.Receiver operating characteristic(ROC)curve and decision curve analysis(DCA)were used to evaluate the diagnostic efficacy and clinical value of the model.Results The model based on the radiomics features of the whole brain gray matter(GM)and the left globus pallidus(LGP)had better diagnostic efficacy and clinical application value.The area under the curve(AUC)in the training set and the test set were 0.813,0.815 and 0.806,0.806,respectively.Conclusion The GM-based radiomics model can effectively diagnose cerebral damage following CO poisoning.

Objective:To establish a method for detecting serum non-ceruloplasmin-bound copper (NCC) by immunoprecipitation-inductively coupled plasma mass spectrometry (IP-ICP-MS) and evaluate its clinical application.Methods:Methodological evaluation research. Immunoprecipitation was first used to separate serum ceruloplasmin, followed by detection of serum NCC levels using ICP-MS. Two levels of quality control serum were reconstituted to determine the limit of detection (LOD), limit of quantitation (LOQ), accuracy, and precision of the self-developed method. The serum samples of 131 healthy individuals (healthy group) and 69 first-time diagnosed Wilson′s disease (WD) patients(WD group) from November 2023 to June 2024 in the Affiliated Hospital of Institute of Neurology, Anhui University of Traditional Chinese Medicine were collected. Using self-developed method detected the serum NCC levels, established the healthy reference intervals, and NCC levels between the two groups were compared using the Mann-Whitney U test. Results:The calibration curve exhibited excellent lineary across the concentration range of 1.562 to 300.000 μg/L, as demonstrated by coefficient of determination ( R2) of 0.999. The self-developed NCC method exhibited that the LOD was 0.99 μg/L, the LOQ was 3.29 μg/L, the accuracy (spike and recovery experience) was 87.67%?106.27%, and the intra-batch and inter-batch imprecision expressed as coefficient of variation ( CV) was 2.8%?7.3%, and the healthy reference range was 34.31-71.79 μg/L. The serum NCC levels in the WD group were 102.39 (74.38, 144.04) μg/L, which was significantly higher than those in healthy group (51.45±10.34) μg/L ( Z=?7.967, P<0.01). Conclusions:The established IP-ICP-MS method for detecting serum NCC meets the required analytical performance criteria. It is simply operative, highly sensitive, and provides accurate and reliable results, which could be used for clinical detection.

The management of antibiotic-resistant, bacterial biofilm infections in skin wounds poses an increasingly challenging clinical scenario. Pseudomonas aeruginosa infection is difficult to eradicate because of biofilm formation and antibiotic resistance. In this study, we identified a new benzothiazole derivative compound, SN12 (IC₅₀ = 43.3 nmol/L), demonstrating remarkable biofilm inhibition at nanomolar concentrations in vitro. In further activity assays and mechanistic studies, we formulated an unconventional strategy for combating P. aeruginosa-derived infections by targeting the two-component (Gac/Rsm) system. Furthermore, SN12 slowed the development of ciprofloxacin and tobramycin resistance. By using murine skin wound infection models, we observed that SN12 significantly augmented the antibacterial effects of three widely used antibiotics-tobramycin (100-fold), vancomycin (200-fold), and ciprofloxacin (1000-fold)-compared with single-dose antibiotic treatments for P. aeruginosa infection in vivo. The findings of this study suggest the potential of SN12 as a promising antibacterial synergist, highlighting the effectiveness of targeting the two-component system in treating challenging bacterial biofilm infections in humans.

Objective:To explore the dilemmas and demands encountered by enterostomal therapists in providing sexual education or guidance to young and middle-aged patients with enterostomy.Methods:From April to October 2022, purposive sampling was used to select 15 enterostomal therapists who received training at the International School of Stomatologists for semi-structured interviews. Colaizzi 7-step analysis method was used for data analysis.Results:Three themes were extracted, namely the "negative behavior" and "positive willingness" of enterostomal therapists, the multiple dilemmas in implementing sex education for young and middle-aged patients with enterostomy, and the demands for sexual education for young and middle-aged patients with enterostomy.Conclusions:Enterostomal therapists should actively pay attention to the sexual education for young and middle-aged patients with enterostomy, overcome multiple obstacles that affect the lack of sexual education for young and middle-aged patients with enterostomy, and persist in promoting sexual education for young and middle-aged patients with enterostomy in clinical practice, in order to improve the marital relationship and quality of marriage between patients and their spouses.

ObjectiveThe lung mesenchymal stem cells （LMSCs） induced by D-galactose （D-gal） were intervened by Wenfei Huaxian decoction-containing serum to explore the mechanism of Wenfei Huaxian decoction in delaying the senescence of LMSCs through the nicotinamide phosphoribosyltransferase/silent information regulator 1 （NAMPT/SIRT1） signaling pathway. MethodWenfei Huaxian decoction-containing serum was prepared. LMSCs were isolated by gradient density centrifugation， and they were cultured and identified in vitro. The senescence model in vitro was established by stimulating cells via D-gal for 24 h. LMSCs cells were modeled after being treated with different volume fractions （5%， 10%， 20%， 40%， and 80%） of Wenfei Huaxian decoction-containing serum for 24 h， and the cell proliferation level was detected by methyl thiazolyl tetrazolium （MTT） method. The cells were randomly divided into blank serum group， model group， and high， medium， and low dose groups of Wenfei Huaxian decoction-containing serum. Senescence-associated β-galactosidase （SA-β-gal） staining was used to detect the senescence of LMSCs in each group. The content of NAD + was detected by colorimetry. The levels of senescence-associated factors （p16 and p53）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in cell culture supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the relative expression of senescence-associated proteins and NAMPT/SIRT1 signaling pathway-related proteins. ResultCompared with the blank serum group， the proliferation of LMSCs was significantly inhibited after D-gal stimulation for 24 h （P<0.01）. Compared with the model group， the proliferation of LMSCs could be promoted after intervention with the corresponding Wenfei Huaxian decoction-containing serum （P<0.05， P<0.01）. Compared with the blank serum group， the SA-β-gal staining of LMSCs in the model group after D-gal stimulation was enhanced， and the content of NAD⁺ was increased （P<0.01）. The expression levels of senescence factors p16 and p53， as well as SASP pro-inflammatory factors IL-6 and TNF-α in the cell culture supernatant， were significantly increased （P<0.01）. The expression of senescence-associated proteins p16， p21， and p53 increased （P<0.01）， and the protein expression of NAMPT， SIRT1， peroxisome proliferator-activated receptor γ coactivator 1α （PGC-1α）， and forkhead box family transcription factor O1 （FoxO1） decreased （P<0.01）. Compared with the model group， the SA-β-gal staining of LMSCs in each group of Wenfei Huaxian decoction-containing serum was significantly reduced， and the content of NAD⁺ was decreased （P<0.01）. The senescence factors （p16 and p53） and inflammatory factors （IL-6 and TNF-α） in the cell culture supernatant were significantly decreased （P<0.01）. The expression of senescence-associated proteins （P16， P21， and P53） decreased （P<0.05， P<0.01）. The protein expressions of NAMPT， SIRT1， PGC-1α， and FoxO1 were significantly up-regulated （P<0.05， P<0.01）. ConclusionWenfei Huaxian decoction can alleviate senescence and inflammatory response damage of D-gal-induced LMSCs， and its mechanism may be related to the regulation of the NAMPT/SIRT1 signaling pathway.

Objective:To explore the clinical effect of using great toe fibular flaps of both feet on reconstruction of pulp defects of two neighbouring digits.Methods:A total of 14 digit pulp defects in 7 cases were repaired in Zhoukou Huaihai Hospital using great toe fibular flaps of both feet from August 2020 to January 2023. Of the 7 cases, there were 4 males and 3 females, with an average of 28 years old, ranging from 19 to 45 years old. Meanwhile, there were 4 cases in left hand and 3 cases in right hand. There were 3 cases of digit pulp defects in index and middle fingers, 2 in middle and ring fingers, and 2 in thumb and index fingers. The area of soft tissue defect in 1.2 cm×1.5 cm-3.0 cm×2.5 cm, and flap was 1.5 cm ×1.8 cm-3.2 cm×2.8 cm. Furthermore, 1 case underwent emergency surgery and 5 were repaired in elective surgery. The donor site of the flap was closed directly, and an intermediate-thickness skin graft was prepared from the medial plantar area for transfer in the case of high suture tension at the wound edge. After surgery, patients received postoperative by outpatient clinic and WeChat to observe the appearance, sensation, functional recovery and flap contracture of digits, as well as the movement of the great toes of both feet.Results:After the surgery, all flaps in the 7 cases survived smoothly and the donor sites healed. All patients entered scheduled follow-ups postoperatively for 6 months to 2 years, with an average of 9 months. The flap showed an aesthetic appearance and excellent sensation, with a TPD of 3-6 mm, and satisfactory digit function. The donor site of the great toe fibular flap left linear scars only, without abnormality in range of motion and gait in walking. In addition, there were 5 in excellent and 2 in good according to the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association.Conclusion:Application of great toe fibular flaps of both feet is an ideal option for the simultaneous repair of pulp defects of two neighbouring digits, which can achieve good reconstructive results.