OBJECTIVES: To elucidate the underlying mechanisms of macrophage-mediated inflammation and tissue injury in diabetic foot ulcer (DFU). METHODS: Skin tissue samples were collected from patients with DFU and with non-DFU. A total of 79 272 high-quality cell transcriptomes were obtained using single-cell RNA sequencing. An unbiased clustering approach was employed to identify cell subpopulations. Seurat functions were used to identify differentially expressed genes between DFU and non-DFU groups, and gene ontology (GO) enrichment analysis was used to reveal gene function. Furthermore, cell-cell communication network construction and ligand-receptor interaction analysis were performed to reveal the mechanisms underlying cellular interactions and signaling regulation in the DFU microenvironment from multiple perspectives. RESULTS: The results revealed a significant expansion of myeloid cells in DFU tissues, alongside a marked reduction in structural cells such as endothelial cells, epithelial cells, and smooth muscle cells. Major cell types underwent functional reprogramming, characterized by immune activation and impaired tissue remodeling. Specifically, macrophages in DFU skin tissues exhibited a shift toward a pro-inflammatory M1 phenotype, with upregulation of genes associated with inflammation and oxidative stress. Cell communication analysis further demonstrated that M1 macrophages served as both primary signal receivers and influencers in the COMPLEMENT pathway mediated communication network, and as key signal senders and mediators in the secreted phosphoprotein 1 (SPP1) pathway mediated communication network, actively shaping the inflammatory microenvironment. Key ligand-receptor interactions driving macrophage signaling were identified, including C3-(ITGAM+ITGB2) and SPP1-CD44. CONCLUSIONS This study establishes a comprehensive single-cell atlas of DFU, revealing the role of macrophage-driven cellular networks in chronic inflammation and impaired healing. These findings may offer potential novel therapeutic targets for DFU treatment.
Humans ; Macrophages/immunology* ; Diabetic Foot/pathology* ; Single-Cell Analysis ; Transcriptome ; Gene Expression Profiling ; Inflammation ; Skin ; Cell Communication ; Signal Transduction ; Cellular Reprogramming

Objective To establish a method for the simultaneous determination of 16 cephalosporin antibiotics of the fourth generation in whole blood by ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS),including representative drugs such as cefalexin,cefuroxime axetil,cefetamet pivoxil,ceftizoxime,cefodizime,cefteram pivoxil,cefpodoxime proxetil,cefditoren pivoxil,cefminox sodium,cefoperazone,cefpirome,cefoxitin,cefamandole nafate,cefquinome sulfate,cefpiramide,and ceftiofur.Methods Whole blood was pretreated with acetonitrile for protein precipitation and then determined by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry.The liquid phase used a Hypersil GOLD? C18 column(2.1 mm ×100 mm,1.9 μm).The organic phase was 0.1％formic acid methanol solution,and the aqueous phase was 0.1％formic acid aqueous solution(containing 10 mmol/mL ammonium formate)for gradient elution.Detection was performed in electrospray positive ionization mode with selected reaction monitoring(SRM).Results The 16 drugs showed good linearity within their respective concentration ranges,with R2 values all greater than 0.99.Limits of detection for cefminox sodium and cefpiramide were 50 and 20 ng/mL,respectively,and for the remaining 14 drugs were all lower than 5 ng/mL.The relative standard deviations(RSDs)of intra-day and inter-day precisions at four spiked concentrations for the 16 drugs were all no higher than 10％(n=5).Accuracy ranged within±15％for mosg drugs,except for cefamandole nafate,ceftiofur,and cefetamet pivoxil at the lower limit of quantification,which showed accuracy within±20％.Extraction recoveries exceeded 80％for all compounds.Conclusion This method has high detection sensitivity,rapid speed,and good repeatability for the simultaneously determination of 16 cephalosporin antibiotics in whole blood.

Objective To explore the characteristics of changes in blood test indicators and the combined detection for the differential diagnosis value of early gastric cancer and atrophic gastritis.Methods A total of 44 patients with early gastric cancer admitted to the hospital from January 2023 to March 2024 were selected as the early gastric cancer group.Another 59 patients with atrophic gastritis who were admitted to the hospital during the same period were selected as the atrophic gastritis group.Analyze the characteristics of changes in blood test indicators of the two groups.The receiver operating characteristic(ROC)curve was drawn to eval-uate the clinical value of blood test indicators in differentiating early gastric cancer from atrophic gastritis.Re-sults The levels of white blood cell count(WBC),platelet count(PLT),C-reactive protein(CRP),pro-thrombin time(PT),fibrinogen(Fib),D-dimer(D-D),and carbohydrate antigen 199(CA199)in the early gastric cancer group were significantly higher than those in the atrophic gastritis group,and the differences were statistically significant(P<0.05).The levels of red blood cells(RBC),hemoglobin(HGB),total pro-tein(TP),thrombin time(TT),and ferritin(Fer)in the early gastric cancer group were significantly lower than those in the atrophic gastritis group,and the difference was statistically significant(P<0.05).The re-sults of the ROC curve analysis show that the area under the curve(AUC)for the individual detection and dif-ferential diagnosis of early gastric cancer and atrophic gastritis by WBC,RBC,HGB,PLT,CRP,TP,PT,TT,Fib,D-D,Fer and CA199 were 0.591,0.716,0.764,0.623,0.728,0.671,0.692,0.746,0.721,0.689,0.770 and 0.680,respectively.The AUC of the combined detection of RBC,HGB,CRP,TT,Fib and Fer for the dif-ferential diagnosis of early gastric cancer and atrophic gastritis was 0.857(95%CI:0.772-0.941),which was significantly higher than AUC of the individual detection.Conclusion The combined detection of blood test indicators such as RBC,HGB,CRP,TT,Fib and Fer has certain clinical value in the differential diagnosis of early gastric cancer and atrophic gastritis.

Objective:To compare the surface electromyography characteristics of essential tremor (ET), Parkinson′s disease (PD) and essential tremor-Parkinson′s disease syndrome (ET-PD).Methods:A total of 74 patients [ET group ( n=23), PD group ( n=30), and ET-PD group ( n=21)] admitted to the Parkinson′s and Movement Disorders Center of the Department of Neurology of Xuanwu Hospital from August 2020 to December 2023 were enrolled, and results of their bilateral upper limbs surface electromyography (sEMG) were collected. sEMG activities were analyzed offline. Power spectral analysis was performed to explore the sEMG activities. One-way analysis of variance and chi-square test were used to compare the differences of electrophysiological parameters. Results:For the ET group characterized by postural tremor, the frequency was (6.06±0.68) Hz, the amplitude was (1 200.91±360.69) μV, the proportion of alternating contractions was 30.4% (7/23), and the proportion of harmonic resonances was 34.8% (8/23). For the PD group characterized by rest tremor, the frequency was (4.81±0.61) Hz, the amplitude was (1 057.40±354.52) μV, the proportion of alternating contractions was 73.3% (22/30), and the proportion of harmonic resonances was 70.0% (21/30). For the ET-PD group in rest and postural state respectively, the frequencies were (5.04±0.44) Hz and (5.80±0.47) Hz, the amplitudes were (1 026.05±191.90) μV and (1 196.67±212.12) μV, the proportions of alternating contractions were 52.4% (11/21) and 38.1% (8/21), and the proportions of harmonic resonances were 52.4% (11/21) and 33.3% (7/21). Analysis of variance revealed that tremor frequencies for the PD group and the ET-PD group in rest state were lower than the ET group and the ET-PD group in postural state ( F=27.439, P<0.001). The proportion of alternating contractions for the PD group was higher than the ET group ( χ 2=9.669, P=0.002) and the ET-PD group in postural state ( χ 2=6.333, P=0.012). The proportion of harmonic resonances for the PD group was higher than the ET group ( χ 2=6.517, P=0.011) and the ET-PD group in postural state ( χ 2=6.708, P=0.010). No statistically significant differences were found for tremor amplitudes among all the groups ( F=2.143, P=0.100). Conclusions:ET is characterized by postural tremors, with a higher frequency and a lower alternating contractions and harmonic resonances. PD is characterized by rest tremors, with a lower frequency and a higher alternating contractions and harmonic resonances. The parameters of ET-PD are between ET and PD, which provide objective evidences for differential diagnosis of tremors.

Objective To establish a method for the simultaneous determination of 16 cephalosporin antibiotics of the fourth generation in whole blood by ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS),including representative drugs such as cefalexin,cefuroxime axetil,cefetamet pivoxil,ceftizoxime,cefodizime,cefteram pivoxil,cefpodoxime proxetil,cefditoren pivoxil,cefminox sodium,cefoperazone,cefpirome,cefoxitin,cefamandole nafate,cefquinome sulfate,cefpiramide,and ceftiofur.Methods Whole blood was pretreated with acetonitrile for protein precipitation and then determined by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry.The liquid phase used a Hypersil GOLD? C18 column(2.1 mm ×100 mm,1.9 μm).The organic phase was 0.1％formic acid methanol solution,and the aqueous phase was 0.1％formic acid aqueous solution(containing 10 mmol/mL ammonium formate)for gradient elution.Detection was performed in electrospray positive ionization mode with selected reaction monitoring(SRM).Results The 16 drugs showed good linearity within their respective concentration ranges,with R2 values all greater than 0.99.Limits of detection for cefminox sodium and cefpiramide were 50 and 20 ng/mL,respectively,and for the remaining 14 drugs were all lower than 5 ng/mL.The relative standard deviations(RSDs)of intra-day and inter-day precisions at four spiked concentrations for the 16 drugs were all no higher than 10％(n=5).Accuracy ranged within±15％for mosg drugs,except for cefamandole nafate,ceftiofur,and cefetamet pivoxil at the lower limit of quantification,which showed accuracy within±20％.Extraction recoveries exceeded 80％for all compounds.Conclusion This method has high detection sensitivity,rapid speed,and good repeatability for the simultaneously determination of 16 cephalosporin antibiotics in whole blood.

Objective:To compare the surface electromyography characteristics of essential tremor (ET), Parkinson′s disease (PD) and essential tremor-Parkinson′s disease syndrome (ET-PD).Methods:A total of 74 patients [ET group ( n=23), PD group ( n=30), and ET-PD group ( n=21)] admitted to the Parkinson′s and Movement Disorders Center of the Department of Neurology of Xuanwu Hospital from August 2020 to December 2023 were enrolled, and results of their bilateral upper limbs surface electromyography (sEMG) were collected. sEMG activities were analyzed offline. Power spectral analysis was performed to explore the sEMG activities. One-way analysis of variance and chi-square test were used to compare the differences of electrophysiological parameters. Results:For the ET group characterized by postural tremor, the frequency was (6.06±0.68) Hz, the amplitude was (1 200.91±360.69) μV, the proportion of alternating contractions was 30.4% (7/23), and the proportion of harmonic resonances was 34.8% (8/23). For the PD group characterized by rest tremor, the frequency was (4.81±0.61) Hz, the amplitude was (1 057.40±354.52) μV, the proportion of alternating contractions was 73.3% (22/30), and the proportion of harmonic resonances was 70.0% (21/30). For the ET-PD group in rest and postural state respectively, the frequencies were (5.04±0.44) Hz and (5.80±0.47) Hz, the amplitudes were (1 026.05±191.90) μV and (1 196.67±212.12) μV, the proportions of alternating contractions were 52.4% (11/21) and 38.1% (8/21), and the proportions of harmonic resonances were 52.4% (11/21) and 33.3% (7/21). Analysis of variance revealed that tremor frequencies for the PD group and the ET-PD group in rest state were lower than the ET group and the ET-PD group in postural state ( F=27.439, P<0.001). The proportion of alternating contractions for the PD group was higher than the ET group ( χ 2=9.669, P=0.002) and the ET-PD group in postural state ( χ 2=6.333, P=0.012). The proportion of harmonic resonances for the PD group was higher than the ET group ( χ 2=6.517, P=0.011) and the ET-PD group in postural state ( χ 2=6.708, P=0.010). No statistically significant differences were found for tremor amplitudes among all the groups ( F=2.143, P=0.100). Conclusions:ET is characterized by postural tremors, with a higher frequency and a lower alternating contractions and harmonic resonances. PD is characterized by rest tremors, with a lower frequency and a higher alternating contractions and harmonic resonances. The parameters of ET-PD are between ET and PD, which provide objective evidences for differential diagnosis of tremors.

Background: Biofilms, such as those from Staphylococcus epidermidis, are generally insensitive to traditional antimicrobial agents, making it difficult to inhibit their formation. Although quercetin has excellent antibiofilm effects, its clinical applications are limited by the lack of sustained and targeted release at the site of S. epidermidis infection. Objectives: Polyethylene glycol-quercetin nanoparticles (PQ-NPs)-loaded gelatin-N,Ocarboxymethyl chitosan (N,O-CMCS) composite nanogels were prepared and assessed for the on-demand release potential for reducing S. epidermidis biofilm formation. Methods: The formation mechanism, physicochemical characterization, and antibiofilm activity of PQ-nanogels against S. epidermidis were studied. Results: Physicochemical characterization confirmed that PQ-nanogels had been prepared by the electrostatic interactions between gelatin and N,O-CMCS with sodium tripolyphosphate. The PQ-nanogels exhibited obvious pH and gelatinase-responsive to achieve on-demand release in the micro-environment (pH 5.5 and gelatinase) of S. epidermidis.In addition, PQ-nanogels had excellent antibiofilm activity, and the potential antibiofilm mechanism may enhance its antibiofilm activity by reducing its relative biofilm formation, surface hydrophobicity, exopolysaccharides production, and eDNA production. Conclusions This study will guide the development of the dual responsiveness (pH and gelatinase) of nanogels to achieve on-demand release for reducing S. epidermidis biofilm formation.

Objective:To investigate the predictive value of inflammatory and nutritional indices for postoperative survival of elderly patients with esophageal squamous carcinoma.Methods:The retrospective cohort study was conducted. The clinicopathological data of 130 elderly patients with esophageal squamous carcinoma who were admitted to Sichuan Cancer Hospital from January 2019 to April 2020 were collected. There were 102 males and 28 females, aged (70±4)years. Mea-surement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represented as M( Q1, Q3) or M(range). Count data were expressed as absolute numbers, and comparison between groups was conducted using the chi-square test. Receiver opera-ting characteristic (ROC) curves were plotted. The area under the curve (AUC) and optimal cut-off values were calculated. The Kaplan-Meier method was used to plot survival curves, and the Log-rank test was used for survival analysis. The COX proportional hazard regression model was used for univariate and multivariate analyses. Results:(1) Postoperative survival of elderly patients with esophageal squamous carcinoma predicted by inflammatory and multitional indices. Results of ROC curves analysis showed that the best cut-off values of preoperative systemic immune-inflammation index (SII), systemic inflammation response index (SIRI), neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and prognostic nutrition index (PNI) for predicting postoperative survival of elderly patients with esophageal squamous carcinoma were 470.71×10 9/L, 1.11, 2.07, 136.24, and 46.28, respectively. (2) Risk factors analysis of postoperative survival of elderly patients with esophageal squamous carcinoma. Results of multivariate analysis showed that preoperative SII ≥470.71×10 9/L, preoperative SIRI ≥1.11, preoperative PNI<46.28, score of preoperative patient-generated subjective global assessment (PG-SGA) ≥4, postoperative pathological stage Ⅳ and post-operative complications were independent risk factors for the overall survival time of elderly patients with esophageal squamous carcinoma ( hazard ratio=3.30, 2.50, 0.36, 4.86, 1.57, 1.97, 95% confidence interval as 1.10?9.88, 1.07?5.88, 0.16?0.81, 1.13?20.87, 1.20?2.06, 1.02?3.82, P<0.05). (3) Follow-up. All the 130 patients were followed up for 39(range, 1?60)months. Of the 130 patients, 81 cases survived, 49 cases died, and the median overall survival time was not reached. The 1- and 3-year survival rates of the 130 patients were 83.85% and 54.62%, respectively. ① The median overall survival time was 25(0,43)months for patients with SII ≥470.71×10 9/L, and unreached for patients with SII <470.71×10 9/L, showing a significant difference between them ( χ2=60.59, P<0.05). ② The median overall survival time was 26(0,44)months for patients with SIRI ≥1.11, and unreached for patients with SIRI <1.11, showing a significant difference between them ( χ2=45.57, P<0.05). ③ The median overall survival time was unreached for patients with PNI ≥46.28, and 38(0,47)months for patients with PNI <46.28, showing a significant difference between them ( χ2=12.53, P<0.05). ④ The median overall survival time was unreached for patients with PG-SGA <4 and ≥4, showing a signifi-cant difference between them ( χ2=14.41, P<0.05). ⑤ The median overall survival time was 25(1,47)months for patients in pathological stage Ⅲ, 12(1,32)months for patients in stage Ⅳ, and unreached for patients in stage 0, Ⅰ, Ⅱ, respectively, showing a significant difference among them ( χ2=58.75, P<0.05). ⑥ The median overall survival time was 33(1,47)months for patients with postoperative complication, and unreached for patients without postoperative complication, showing a significant difference between them ( χ2=14.27, P<0.05). Conclusions:Preoperative SII, SIRI and PNI have good predictive value for postoperative survival in elderly patients with esophageal squamous carcinoma. Preoperative SII ≥470.71×10 9/L, preoperative SIRI ≥1.11, preoperative PNI <46.28, score of preoperative PG-SGA ≥4, postoperative pathological stage Ⅳ, and postoperative complications are independent risk factors for the overall survival time of elderly patients with esophageal squamous carcinoma. Patients with preoperative SII <470.71×10 9/L, preoperative SIRI <1.11, preoperative PNI >46.28, score of preoperative PG-SGA <4, postoperative pathological stage 0, Ⅰ, Ⅱ, and non post-operative complications have better survival.

Objective To explore the role of serum leucine-rich-alpha-2-glycoprotein1(LRG1),neopterin(NPT)and virus antibodies during the progression of viral meningitis(VM)and their value in evaluating the condition and predicting the prognosis of VM in children.Methods A total of 130 pediatric patients with VM admitted to Kaifeng Children's Hospital from August 2020 to February 2023 were selected as the observation group,and at the same time,another 130 pediatric patients with VM symptoms who were diagnosed as non central nervous system infectious diseases by lumbar puncture cerebrospinal fluid examination were selected as the control group.After admission,2 mL peripheral venous blood of children in the two groups was collected,and the levels of serum LRG1 and NPT were assessed by enzyme-linked immunosorbent assay.Before treatment,1 mL cerebrospinal fluid sample of children in the two groups was collected by lumbar puncture,and the host specific immuno-globulin G(IgG)against herpes simplex virus-I(HSV-I),cytomegalovirus(CMV),Epstein-Barr virus(EBV),and parain-fluenza virus(PIV)in cerebrospinal fluid was detected by enzyme-linked immunosorbent assay.The levels of serum LRG1 and NPT,as well as the positive rate of virus antibody IgG were compared between the two groups.The VM children were then divided into the severe group(n=54)and mild group(n=76)according to the disease severity,and they were divided into the good prognosis group(n=37)and poor prognosis group(n=93)based on the prognosis.The levels of serum IRG1 and NPT and the positive rate of virus antibody IgG were compared in children with different disease severity and prognosis.The correlation of serum LRG1,NPT and virus antibody with the disease severity or disease progression for pediatric VM was analyzed,and their interactive effects on the disease progression of children with VM were analyzed.The predictive value of serum LRG1,NPT and virus antibody for pediatric VM outcome was analyzed by the receiver operating characteristic(ROC)curve.Results The levels of serum LRG1 and NPT,as well as the positive rate of virus antibody IgG of children in the obser-vation group were significantly higher than those in the control group(P＜0.05);the levels of serum LRG1 and NPT,as well as the positive rate of virus antibody IgG of children in the severe group were significantly higher than those in the mild group(P＜0.05);the levels of serum LRG1 and NPT,as well as the positive rate of virus antibody IgG of children in the poor prognosis group were significantly higher than those in the good prognosis group(P＜0.05).High level of serum LRG1 with high positive rate of virus antibody IgG and high level of serum NPT with high positive rate of virus antibody IgG showed positive interactions in leading to poor prognosis in children with VM odds ratio(OR)=15.238,9.684).The submultiplicative models were applied for the high levels of serum LRG1 with high positive rate of virus antibody IgG(OR=15.238)and high levels of serum NPT with high positive rate of virus antibody IgG(OR=9.684).The area under the curve(AUC)for predicting poor prognosis in children with VM after 2 weeks of treatment by serum LRG1,NPT and virus antibody IgG were 0.786,0.794 and 0.919,respectively,and the AUC for the combined prediction of the three was 0.933;the AUC of LRG1,NPT and positive rate of virus antibody IgG for predicting poor prognosis in children with VM after 2 weeks of treatment was greater than that of the three alone(P＜0.05).Conclusion NPT and LRG1 may be involved in the development of VM.The levels of serum LRG1 and NPT are positively correlated with the severity and progression of VM.The levels of serum NPT and LRG1 and the positive rate of virus antibody IgG have certain value in the disease evaluation and prognosis prediction of VM,and the combination of the three has a higher predictive value for the prognosis of VM.There is a positive interaction between high LRG1 with high positive rate of virus antibody IgG,and high NPT with high positive rate of virus antibody IgG in children with VM.

Objective To analyze the distribution and drug resistance patterns of carbapenem-resistant Acinetobacter baumannii(CRAB),and evaluate the antimicrobial effects in vitro of colistin(COL)combined with cefoperazone/sulbactam(SCF),tigecycline(TGC),imipenem(IPM),meropenem(MEM),amikacin(AK),and levofloxacin(LEV)against CRAB to provide guidance for anti-infective therapy.Methods A total of 75 non-duplicate CRAB strains isolated from clinical specimens in 2022 were collected.WHONET 5.6 software was used for retrospective analysis of their clinical distribution and resistance profiles.Among them,25 strains were selected for combined antimicrobial susceptibility tests using broth microdilution to determine minimum inhibitory concentrations(MICs).The fractional inhibitory concentration(FIC)index was calculated based on checkerboard method to assess the combined effects.Results In 2022,a total of 145 Acinetobacter baumannii strains were isolated,including 75 CRAB(detection rate of 51.7％).CRAB was most prevalent in the patients over 70 years old(40.0％),mainly from blood(41.3％)and sputum(37.3％)specimens,and the intensive care unit was the top isolating department.All the 75 CRAB strains were resistant to piperacillin,piperacillin/tazobactam,ceftriaxone,1PM and MEM with resistance rates of 100％.The resistance rates for ampicillin/sulbactam,cefepime and tri-methoprim/sulfamethoxazole were all exceeded 95％.The resistance rates were 86.7％for LEV,82.7％for SCF,77.4％for AK,2.4％for TGC,and 0.0％for COL.The results of combined tests of 25 strains revealed the synergy rates of 92％and 80％for COL+SCF and COL+TGC respectively with sums of both synergy and additive rates of 100％.Synergy rates for COL combined with IPM,MEM,AK and LEV were 64％,72％,56％and 48％respectively.No antagonism was observed in any combination.Conclusion The drug resist-ance of CRAB exhibited high levels at our hospital,and the elderly and ICU patients were the major susceptible populations.Among all the combinations tests,COL+SCF showed optimal synergy while COL+LEV only had the lowest combined effect with the findings that warrants clinical consideration.