Objective:To explore the expression level of Porphyromonas gingivalis, downstream of tyrosine kinase 3 (DOK3) and tumor-associated macrophage (TAM) in the tumor immunomicroenvironment of oral squamous cell carcinoma (OSCC) and the correlation with clinicopathological characteristics and prognosis of patients.Methods:A retrospective case-control study was conducted. The clinical data of 200 OSCC patients with Porphyromonas gingivalis-positive confirmed by 16S rDNA sequencing technology in the First Affiliated Hospital, Xinjiang Medical University between June 2008 and June 2020 were collected. The tumor tissues and the corresponding adjacent normal mucosal tissues of 6 OSCC patients (including 3 cases with Porphyromonas gingivalis -positive and 3 cases with Porphyromonas gingivalis-negative) were selected for high-throughput sequencing to screen differentially co-expressed genes. Immunohistochemistry method was used to detect the expressions of Porphyromonas gingivalis, DOK3, and CD206 (a TAM marker). The median H score of OSCC tissues was used as the threshold to categorize the expression level of Porphyromonas gingivalis, DOK3 and CD206 into low-expression (H score < threshold) and high-expression (H score ≥ threshold) groups. The overall survival (OS) analysis was conducted by using the Kaplan-Meier method, and the log-rank test was employed.Results:The high-throughput sequencing results revealed that DOK3 is a differentially co-expressed gene among normal oral mucosa, Porphyromonas gingivalis-positive, and Porphyromonas gingivalis-negative OSCC. In 200 patients with Porphyromonas gingivalis-positive OSCC, 139 exhibited high expression of Porphyromonas gingivalis (H score ≥ 7 points), while 61 showed low expression (H score < 7 points). There were statistically significant differences in the expression levels of Porphyromonas gingivalis in patients with different survival status, pathological T stage, pathological N stage, clinical stage, tumor diameter, degree of tumor differentiation and recurrence (all P < 0.05). Among the 139 OSCC patients with high expression of Porphyromonas gingivalis, 92 cases showed high expression of DOK3 (H score ≥ 6 points) and 47 showed low expression (H score < 6 points); 78 cases exhibited high expression of CD206 (H score ≥ 6 points), while 61 showed low expression (H score < 6 points). There were statistically significant differences in the DOK3 expression level in the high expression of Porphyromonas gingivalis OSCC patients with different age, survival status, pathological T stage, pathological N stage, and recurrence (all P < 0.05). There were statistically significant differences in the CD206 expression level in the high expression of Porphyromonas gingivalis OSCC patients with different pathological T stage, clinical stage, and degree of tumor differentiation (all P < 0.05). The expression of Porphyromonas gingivalis was positively correlated with the expressions of DOK3 and CD206 (both P < 0.01). At the last follow-up on April 6th, 2024, the median follow-up time was 45 months (3 to 106 month range). The median OS time of the 200 patients was 2 429 d, and the 3-year OS rate was 63.9%. The OS of OSCC patients with high expressions of Porphyromonas gingivalis, DOK3, and CD206 was worse than that in those with low expressions (all P < 0.05). Conclusions:The high expression levels of Porphyromonas gingivalis, DOK3, and TAM are associated with a poor prognosis of OSCC patients, suggesting their potential as key biomarkers for prognostic evaluation.

Champagne bottle neck sign (CBNS) is an important morphological feature of extracranial artery damage in patients with moyamoya disease (MMD), and more than half of them have this sign. Carotid ultrasound is the most convenient imaging examination method for diagnosing CBNS. CBNS can have a serious impact on cerebral hemodynamics and is closely associated with the staging of MMD, stroke risk, stroke characteristics, MMD-related headaches, and the risk of postoperative complications. This article comprehensively reviews the pathology and pathogenesis, imaging examination, correlation with clinical symptoms, and intervention of CBNS in patients with MMD, aiming to provide reference for the clinical diagnosis, treatment, and research of MMD combined with CBNS.

Objective To investigate the changing antibiotic resistance profiles of E.coli isolated from patients in the 52 hospitals participating in the CHINET program from 2015 to 2021.Methods Antimicrobial susceptibility was tested for clinical isolates of E.coli according to the unified protocol of CHINET program.WHONET 5.6 and SPSS 20.0 software were used for data analysis.Results Atotal of 289 760 nonduplicate clinical strains ofE.coli were isolated from 2015 to 2021,mainly from urine samples(44.7±3.2)％.The proportion of E.coli strains isolated from urine samples was higher in females than in males(59.0％vs 29.5％).The proportion of E.coli strains isolated from respiratory tract and cerebrospinal fluid samples was significantly higher in children than in adults(16.7％vs 7.8％,0.8％vs 0.1％,both P＜0.05).The isolates from internal medicine department accounted for the largest proportion(28.9±2.8)％with an increasing trend over years.Overall,the prevalence of ESBLs-producing E.coli and carbapenem resistant E.coli(CREco)was 55.9％and 1.8％,respectively during the 7-year period.The prevalence of ESBLs-producing E.coli was the highest in tertiary hospitals each year from 2015 to 2021 compared to secondary hospitals.The prevalence of CREco was higher in children's hospitals compared to secondary and tertiary hospitals each year from 2015 to 2021.The prevalence of ESBLs-producing E.coli in tertiary hospitals and children's hospitals and the prevalence of CREco in children's hospitals showed a decreasing trend over the 7-year period.The prevalence of CREco in secondary and tertiary hospitals increased slowly.Antibiotic resistance rates changed slowly from 2015 to 2021.Carbapenem drugs(imipenem,meropenem)were the most active drugs amongβ-lactams against E.coli(resistance rate≤2.1％).The resistance rates of E.coli to β-lactam/β-lactam inhibitor combinations(piperacillin-tazobactam,cefoperazone-sulbactam),aminoglycosides(amikacin),nitrofurantoin and fosfomycin(for urinary isolates only)were all less than 10％.The resistance rate of E.coli strains to antibiotics varied with the level of hospitals and the departments where the strains were isolated,especially for cefazolin and ciprofloxacin,to which the resistance rate of E.coli strains from children in non-ICU departments was significantly lower than that of the strains isolated from other departments(P＜0.05).The E.coli isolates from ICU showed higher resistance rate to most antimicrobial agents tested(excluding tigecycline)than the strains isolated from other departments.The E.coli strains isolated from tertiary hospitals showed higher resistance rates to the antimicrobial agents tested(excluding tigecycline,polymyxin B,cefepime and carbapenems)than the strains from secondary hospitals and children's hospitals.Conclusions E.coli is an important pathogen causing clinical infection.More than half of the clinical isolates produced ESBL.The prevalence of CREco is increasing in secondary and tertiary hospitals over the 7-year period even though the overall prevalence is still low.This is an issue of concern.

Immobilized enzyme-based enzyme electrode biosensors, characterized by high sensitivity and efficiency, strong specificity, and compact size, demonstrate broad application prospects in life science research, disease diagnosis and monitoring, etc. Immobilization of enzyme is a critical step in determining the performance (stability, sensitivity, and reproducibility) of the biosensors. Random immobilization (physical adsorption, covalent cross-linking, etc.) can easily bring about problems, such as decreased enzyme activity and relatively unstable immobilization. Whereas, directional immobilization utilizing amino acid residue mutation, affinity peptide fusion, or nucleotide-specific binding to restrict the orientation of the enzymes provides new possibilities to solve the problems caused by random immobilization. In this paper, the principles, advantages and disadvantages and the application progress of enzyme electrode biosensors of different directional immobilization strategies for enzyme molecular sensing elements by specific amino acids (lysine, histidine, cysteine, unnatural amino acid) with functional groups introduced based on site-specific mutation, affinity peptides (gold binding peptides, carbon binding peptides, carbohydrate binding domains) fused through genetic engineering, and specific binding between nucleotides and target enzymes (proteins) were reviewed, and the application fields, advantages and limitations of various immobilized enzyme interface characterization techniques were discussed, hoping to provide theoretical and technical guidance for the creation of high-performance enzyme sensing elements and the manufacture of enzyme electrode sensors.

Objective To investigate the changing antibiotic resistance profiles of E.coli isolated from patients in the 52 hospitals participating in the CHINET program from 2015 to 2021.Methods Antimicrobial susceptibility was tested for clinical isolates of E.coli according to the unified protocol of CHINET program.WHONET 5.6 and SPSS 20.0 software were used for data analysis.Results Atotal of 289 760 nonduplicate clinical strains ofE.coli were isolated from 2015 to 2021,mainly from urine samples(44.7±3.2)％.The proportion of E.coli strains isolated from urine samples was higher in females than in males(59.0％vs 29.5％).The proportion of E.coli strains isolated from respiratory tract and cerebrospinal fluid samples was significantly higher in children than in adults(16.7％vs 7.8％,0.8％vs 0.1％,both P＜0.05).The isolates from internal medicine department accounted for the largest proportion(28.9±2.8)％with an increasing trend over years.Overall,the prevalence of ESBLs-producing E.coli and carbapenem resistant E.coli(CREco)was 55.9％and 1.8％,respectively during the 7-year period.The prevalence of ESBLs-producing E.coli was the highest in tertiary hospitals each year from 2015 to 2021 compared to secondary hospitals.The prevalence of CREco was higher in children's hospitals compared to secondary and tertiary hospitals each year from 2015 to 2021.The prevalence of ESBLs-producing E.coli in tertiary hospitals and children's hospitals and the prevalence of CREco in children's hospitals showed a decreasing trend over the 7-year period.The prevalence of CREco in secondary and tertiary hospitals increased slowly.Antibiotic resistance rates changed slowly from 2015 to 2021.Carbapenem drugs(imipenem,meropenem)were the most active drugs amongβ-lactams against E.coli(resistance rate≤2.1％).The resistance rates of E.coli to β-lactam/β-lactam inhibitor combinations(piperacillin-tazobactam,cefoperazone-sulbactam),aminoglycosides(amikacin),nitrofurantoin and fosfomycin(for urinary isolates only)were all less than 10％.The resistance rate of E.coli strains to antibiotics varied with the level of hospitals and the departments where the strains were isolated,especially for cefazolin and ciprofloxacin,to which the resistance rate of E.coli strains from children in non-ICU departments was significantly lower than that of the strains isolated from other departments(P＜0.05).The E.coli isolates from ICU showed higher resistance rate to most antimicrobial agents tested(excluding tigecycline)than the strains isolated from other departments.The E.coli strains isolated from tertiary hospitals showed higher resistance rates to the antimicrobial agents tested(excluding tigecycline,polymyxin B,cefepime and carbapenems)than the strains from secondary hospitals and children's hospitals.Conclusions E.coli is an important pathogen causing clinical infection.More than half of the clinical isolates produced ESBL.The prevalence of CREco is increasing in secondary and tertiary hospitals over the 7-year period even though the overall prevalence is still low.This is an issue of concern.

Objective:To explore the expression level of Porphyromonas gingivalis, downstream of tyrosine kinase 3 (DOK3) and tumor-associated macrophage (TAM) in the tumor immunomicroenvironment of oral squamous cell carcinoma (OSCC) and the correlation with clinicopathological characteristics and prognosis of patients.Methods:A retrospective case-control study was conducted. The clinical data of 200 OSCC patients with Porphyromonas gingivalis-positive confirmed by 16S rDNA sequencing technology in the First Affiliated Hospital, Xinjiang Medical University between June 2008 and June 2020 were collected. The tumor tissues and the corresponding adjacent normal mucosal tissues of 6 OSCC patients (including 3 cases with Porphyromonas gingivalis -positive and 3 cases with Porphyromonas gingivalis-negative) were selected for high-throughput sequencing to screen differentially co-expressed genes. Immunohistochemistry method was used to detect the expressions of Porphyromonas gingivalis, DOK3, and CD206 (a TAM marker). The median H score of OSCC tissues was used as the threshold to categorize the expression level of Porphyromonas gingivalis, DOK3 and CD206 into low-expression (H score < threshold) and high-expression (H score ≥ threshold) groups. The overall survival (OS) analysis was conducted by using the Kaplan-Meier method, and the log-rank test was employed.Results:The high-throughput sequencing results revealed that DOK3 is a differentially co-expressed gene among normal oral mucosa, Porphyromonas gingivalis-positive, and Porphyromonas gingivalis-negative OSCC. In 200 patients with Porphyromonas gingivalis-positive OSCC, 139 exhibited high expression of Porphyromonas gingivalis (H score ≥ 7 points), while 61 showed low expression (H score < 7 points). There were statistically significant differences in the expression levels of Porphyromonas gingivalis in patients with different survival status, pathological T stage, pathological N stage, clinical stage, tumor diameter, degree of tumor differentiation and recurrence (all P < 0.05). Among the 139 OSCC patients with high expression of Porphyromonas gingivalis, 92 cases showed high expression of DOK3 (H score ≥ 6 points) and 47 showed low expression (H score < 6 points); 78 cases exhibited high expression of CD206 (H score ≥ 6 points), while 61 showed low expression (H score < 6 points). There were statistically significant differences in the DOK3 expression level in the high expression of Porphyromonas gingivalis OSCC patients with different age, survival status, pathological T stage, pathological N stage, and recurrence (all P < 0.05). There were statistically significant differences in the CD206 expression level in the high expression of Porphyromonas gingivalis OSCC patients with different pathological T stage, clinical stage, and degree of tumor differentiation (all P < 0.05). The expression of Porphyromonas gingivalis was positively correlated with the expressions of DOK3 and CD206 (both P < 0.01). At the last follow-up on April 6th, 2024, the median follow-up time was 45 months (3 to 106 month range). The median OS time of the 200 patients was 2 429 d, and the 3-year OS rate was 63.9%. The OS of OSCC patients with high expressions of Porphyromonas gingivalis, DOK3, and CD206 was worse than that in those with low expressions (all P < 0.05). Conclusions:The high expression levels of Porphyromonas gingivalis, DOK3, and TAM are associated with a poor prognosis of OSCC patients, suggesting their potential as key biomarkers for prognostic evaluation.

In recent years, the application of minimal residual disease (MRD) in solid tumors has gained widespread attention. MRD typically refers to the presence of residual cancer cells that remain undetectable by imaging after curative treatments, such as surgical resection. The presence of MRD post-surgery is significantly associated with an increased risk of tumor recurrence. In colorectal cancer, circulating tumor DNA (ctDNA) serves as an effective marker for assessing MRD, particularly in non-metastatic (stages I-III) colorectal cancer. As a real-time, accurate, and convenient biomarker, ctDNA can effectively predict tumor recurrence, guide postoperative adjuvant chemotherapy decisions, and provide crucial information for recurrence monitoring. The application prospects of ctDNA detection technology are vast, promising more precise and individualized treatment plans for colorectal cancer patients. This article comprehensively analyzes the progress in the application of ctDNA for detecting MRD in non-metastatic colorectal cancer patients, elaborates on its guiding role in clinical treatment decisions, and envisions the future development directions in this field.

Aim To investigate the intracellular up-take and target protein binding characteristics of two Bruton's tyrosine kinase inhibitors(BTKi)with differ-ent selectivities to provide further insights into the mechanisms of drug off-target-related bleeding risk.Methods Ibrutinib(non-selective BTKi)and za-nubrutinib(selective BTKi)were used as study drugs.After incubation of MEC-1 cells and human platelets with drugs,the cellular thermal shift assay(CETSA)was combined with Western blot to obtain the melting curve and isothermal curve to analyze the binding char-acteristics of the two drugs with the target protein BTK.After incubation of MEC-1 cells and human platelets with drugs,the concentrations of the two drugs were detected by liquid chromatography-tandem mass spectrometry(LC-MS/MS)to analyze the intracellular uptake of the two drugs.Results CETSA analysis confirmed that zanubrutinib was more selective for the target protein BTK compared to ibrutinib.LC-MS/MS analysis showed that both drugs were uptaken intracel-lularly by MEC-1 cells and platelets in a concentration-dependent manner.Conclusions While BTKi targe-ting BTK to B lymphocytes exerts therapeutic effects,off-target effects on platelets due to differences in their intracellular uptake,and target-binding characteristics may be one of the reasons for the differences in bleed-ing risk across selective BTKi.

In recent years, the application of minimal residual disease (MRD) in solid tumors has gained widespread attention. MRD typically refers to the presence of residual cancer cells that remain undetectable by imaging after curative treatments, such as surgical resection. The presence of MRD post-surgery is significantly associated with an increased risk of tumor recurrence. In colorectal cancer, circulating tumor DNA (ctDNA) serves as an effective marker for assessing MRD, particularly in non-metastatic (stages I-III) colorectal cancer. As a real-time, accurate, and convenient biomarker, ctDNA can effectively predict tumor recurrence, guide postoperative adjuvant chemotherapy decisions, and provide crucial information for recurrence monitoring. The application prospects of ctDNA detection technology are vast, promising more precise and individualized treatment plans for colorectal cancer patients. This article comprehensively analyzes the progress in the application of ctDNA for detecting MRD in non-metastatic colorectal cancer patients, elaborates on its guiding role in clinical treatment decisions, and envisions the future development directions in this field.