ObjectiveThe Golgi apparatus serves as a central hub in the eukaryotic secretory pathway, responsible for the processing, sorting, and trafficking of proteins and lipids. In mammalian cells, the Golgi typically forms a perinuclear ribbon-like structure composed of laterally connected cisternae stacks.The maintenance of Golgi ribbon structure depends on the balance of membrane flux across multiple intracellular trafficking pathways, yet the specific contributions of distinct trafficking branches to Golgi macroscopic morphology remain elusive. In mammalian cells, the Golgi ribbon is typically organized as a perinuclear, laterally connected structure composed of stacked cisternae, and its integrity is highly dynamic and sensitive to perturbations in membrane trafficking. This study aims to elucidate the role of coat protein complex I (COPI)-mediated retrograde transport in maintaining the Golgi ribbon and to dissect the functional relationship between the transmembrane cargo receptors LEPROT/LEPROTL1 (LEPROTs) and the COPI adaptor GOLPH3. MethodsUsing siRNA interference and gene-deficient cell lines, we selectively perturbed COPI- or adaptor protein complex 1 (AP-1)-mediated trafficking pathways in HeLa cells. To quantitatively evaluate Golgi morphology, we employed a “Golgi Angle”-based measurement to assess its circumferential distribution around the nucleus. The spatial distribution of the Golgi ribbon was quantitatively analyzed using confocal microscopy, while Golgi ultrastructure and vesicle density were examined via transmission electron microscopy. Additionally, the subcellular distribution of COPI components was assessed by immunofluorescence co-localization. ResultsSelective inhibition of COPI retrograde transport significantly induced the circumferential extension of the Golgi ribbon around the nucleus, whereas blocking AP-1-mediated anterograde transport resulted in Golgi compaction, indicating opposing roles. These results suggest that different trafficking branches downstream of ARF1 exert distinct and even antagonistic effects on Golgi morphology. LEPROTs-deficient cells exhibited a Golgi extension phenotype highly consistent with COPI impairment. Furthermore, knockdown of GOLPH3 in a LEPROTs double-knockout background produced a significant additive effect on Golgi extension, suggesting that LEPROTs and GOLPH3 play non-redundant roles in regulating COPI-related trafficking processes. Mechanistically, loss of either LEPROTs or GOLPH3 led to the aberrant accumulation of COPI components at endoplasmic reticulum exit sites, accompanied by a reduction in COPI-like vesicles around the Golgi. This redistribution indicates a defect in COPI recycling between the ER-Golgi interface and the Golgi apparatus. Ultrastructural analysis revealed that Golgi cisternae in defective cells became shorter and thicker while maintaining a stable number of stacks. In parallel, the density of Golgi-associated vesicles was markedly decreased, further supporting an impairment in COPI vesicle formation or budding processes. ConclusionThis study demonstrates that active COPI retrograde transport is a critical factor in restricting the over-connection of the Golgi ribbon and maintaining its compactness. Rather than causing fragmentation, partial disruption of COPI function leads to a distinct morphological outcome characterized by Golgi ribbon extension at the light microscopy level and cisternal remodeling at the ultrastructural level. LEPROTs and GOLPH3 cooperatively promote the recycling and vesiculation of COPI components, thereby imposing a structural constraint on the Golgi periphery. Our findings support a model in which multiple adaptor proteins act in parallel to sustain efficient COPI cycling, thereby maintaining Golgi structural homeostasis. These findings provide new cell biological evidence for the membrane trafficking basis of Golgi morphological homeostasis.

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.

ObjectiveIn order to provide a reference for the optimization of preparation process of stir-fried Glycyrrhizae Radix et Rhizoma（sf-GRR）， the quality changes during the processing was studied. MethodsGlycyrrhizae Radix et Rhizoma was processed by stir-frying for 17 min， and samples were collected every 1 min during the processing. The appearance color of the samples was determined by visual analysis technology， the moisture and extract of the process samples were detected by the drying method and the hot extraction method of alcohol-soluble extract in the general rules of the 2020 edition of Chinese Pharmacopoeia（part Ⅳ）， and the contents of liquiritin apioside， liquiritin， isoliquiritin apioside， isoliquiritin， licoricesaponin G₂ and glycyrrhizic acid in the process samples were determined by high performance liquid chromatography（HPLC）. Then principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and Spearman correlation analysis were used for clustering， discrimination and correlation analysis of the appearance color， moisture， extract and the contents of six internal components. Based on artificial neural network and random forest algorithm， the prediction model of processing degree of sf-GRR was established. On this basis， based on the five principles of quality marker（Q-Maker）， explore the monitoring Q-Maker of sf-GRR. ResultsThe color of Glycyrrhizae Radix et Rhizoma deepened after stir-frying， and the appearance color of the sample changed from light yellow to dark yellow during processing. During the stir-frying process， the moisture content showed a decreasing trend with the extension of processing time， while the extract content showed an increasing trend with the extension of processing time. After stir-frying， the contents of liquiritin apioside， liquiritin and licoricesaponin G₂ showed an overall decreasing trend， while the contents of isoliquiritin apioside and isoliquiritin increased， and the content of glycyrrhizic acid increased slightly. The correlation analysis showed that moisture was positively correlated with brightness（L^*） and red/green value（a^*）， and negatively correlated with yellow/blue value（b^*） and total color difference（E^*ab）. Isoliquiritin apioside and isoliquiritin had negative correlation with L^* and a^*， and positive correlation with b^* and E^*ab. The processing process of sf-GRR could be divided into two stages of the early stage（0-14 min） and the late stage（15-17 min）， and could be divided into three stages of the early stage（0-6 min）， the middle stage（7-14 min） and the late stage（15-17 min） by combining the moisture， extract， the contents of 6 components and color values. Based on artificial neural network analysis and random forest algorithm， isoliquiritin apioside， isoliquiritin， liquiritin and glycyrrhizic acid were selected as monitoring markers for sf-GRR. ConclusionBased on the analysis of the exterior-interior indicators of process samples of sf-GRR， this paper ultimately identifies four processing monitoring markers， which can provide a basis for optimizing the processing technology of sf-GRR.

Objective:To explore the application value of machine learning prediction model for neural invasion in gallbladder cancer based on enhanced computed tomography (CT) and clinical characteristics.Methods:The retrospective cohort study was conducted. The clinical and imaging data of 502 patients with gallbladder cancer who were admitted to The First Affiliated Hospital of Xinxiang Medical University from January 2010 to June 2024 were collected. There were 171 males and 331 females, aged 65(range, 35?91)years. All patients underwent preoperative abdominal enhanced CT and radical resection. The 502 patients were randomly divided into a training set of 351 cases and a test set of 151 cases at a 7:3 ratio. The training set was used to construct prediction model, and the test set was used to validate prediction model. Observation indicators: (1)neural invasion in gallbladder cancer and influencing factor analysis; (2) construction and validation of machine learning prediction models for neural invasion in gallbladder cancer. Comparison of count data between groups was conducted using the chi-square test. Comparison of ordinal data between groups was conducted using the Mann-Whitney U test. Logistic regression model was performed for univariate and multivariate analyses. Independent influencing factors were incor-porated to construct machine learning models using the standard library modules based on Python 3.9. Receiver operating characteristic (ROC) curves were plotted, and the accuracy, sensitivity, specificity, area under the curve (AUC), precision, F1 score, positive predictive value, negative predic-tive value, and Kappa value were calculated to evaluate the predictive performance of the models. The Delong test was used to assess the differences in AUC among different models in the test set. The Hosmer-Lemeshow test and Brier score were used to evaluate the calibration of the models. Results:(1) Neural invasion in gallbladder cancer and influencing factor analysis. Of the 502 patients with gallbladder cancer, 131 cases had neural invasion, and 371 cases had no neural invasion. Results of multivariate analysis showed that total bilirubin, carcinoembryonic antigen, CA199, CA125, neutrophil-lymphocyte ratio, liver invasion detected by CT, vascular invasion detected by CT, hilar or retroperi-toneal lymph node metastasis detected by CT, and tumor stages T3 and T4 were independent influencing factors for neural invasion in patients with gallbladder cancer [ odds ratios=3.747, 2.395, 3.917, 3.596, 2.805, 2.377, 3.523, 2.774, 5.080, 6.809, 95% confidence interval ( CI) as 1.890?7.430, 1.154?4.971, 2.054?7.472, 1.807?7.155, 1.506?5.225, 1.241?4.553, 1.666?7.449, 1.483?5.189, 2.050?12.589, 2.552?18.168, P<0.05]. (2) Construction and validation of machine learning predic-tion models for neural invasion in gallbladder cancer. Based on the independent influencing factors, seven machine learning models were constructed, including logistic regression, K-nearest neighbors, support vector machine, random forest, decision tree, back-propagation neural network, and gradient boosting machine. The ROC curves of seven machine learning models in the test set were plotted, and the AUC were 0.900(95% CI as 0.851?0.948), 0.741(95% CI as 0.646?0.829), 0.836(95% CI as 0.762?0.895), 0.782(95% CI as 0.701?0.855), 0.839(95% CI as 0.770?0.901), 0.817(95% CI as 0.738?0.887), 0.843(95% CI as 0.770?0.909), respectively. Results of Delong test showed that the logistic regression model had the highest AUC. The sensitivity and specificity of the logistic regression model were 0.868 and 0.805 respectively, indicating the best balance. Results of Hosmer-Lemeshow test showed that the logistic regression model had a good goodness-of-fit ( χ2=5.320, P>0.05). The Brier score of the logistic regression model was relatively low, as 0.168, which verified its calibration advantage. Conclusion:Total bilirubin, carcinoembryonic antigen, CA199, CA125, neutrophil-to-lymphocyte ratio, liver invasion detected by enhanced CT, vascular invasion detected by enhanced CT, hilar or retroperitoneal lymph node metastasis detected by enhanced CT, and tumor stages T3 and T4 are independent influencing factors for nerve invasion in patients with gallbladder cancer. Seven machine learning models are constructed based on enhanced CT and clinical characteristics to predict neural invasion in gallbladder cancer, of which the logistic regression model demonstrates good predictive performance.

Objective To investigate the cIinicopathological features and immunohistochemical expression of gastric hyperplastic polyps(GHPs)with dysplasia/adenocarcinoma.Methods A retrospective analysis of 24 cases(44 polyps)that were diagnosed as GHPs with dysplasia/adenocarcinoma in our hospital from January 2020 to December 2024 was reviewed,and clinical,histomorphological,immunophenotypic and follow-up data were analyzed.Results There were 20 female and 4 male cases,with a mean age of(65.5±7.9)(range 56～76)years.Among 44 polyps,3 occurred in the antrum of the stomach,1 in the gastric horn,and 40 in the fundus/body.Among the polyps,32 cases were diagnosed as high-grade dysplasia,4 cases as low-grade dysplasia,4 cases as coexistence of low-grade+high-grade dysplasia,2 cases as mucinous adenocarcinoma,1 cases as poorly differentiated adenocarcinoma,and 1 cases as signet-ring cell carcinoma.The histological manifestations of 23 cases of background mucosa were autoimmune metaplastic atrophic gastritis(AMAG).the P53 of 8 polyps showed a mutant expression pattern.Through MUC5/MUC6/MUC2/CD10 joint examination,33 cases showed gastric type(25 cases of which were foveal epithelium type),4 cases were intestinal type,5 cases were mixed gastrointestinal type,and 2 cases were non-gastrointestinal type.Conclusion The neoplastic transformation of GHPs is closely related to AMAG.It is necessary for clinicians and pathologists to strengthen their evaluation of background mucosa,to achieve early detection,early diagnosis and early treatment.

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.

In recent decades,the incidence of human papillomavirus(HPV)-associated oropharyngeal squamous cell carcinoma(OPSCC)has shown a marked increase.Significant changes have also occurred in the OPSCC diagnosis and treatment paradigm.Deter-mining HPV status prior to treatment is now essential,and radiotherapy/chemotherapy,immunotherapy,and minimally invasive surgical techniques have progressively emerged as key modalities for managing OPSCC.However,alongside these paradigm shifts,a comprehen-sive technical consensus guiding the entire diagnostic and therapeutic process for OPSCC patients is currently lacking.Given China's large population base and the rising incidence of OPSCC,an expert panel convened to develop a clinical technical consensus on OPSCC diagno-sis and management tailored to China's specific context.This consensus aims to further enhance and standardize understanding of OPSCC management techniques among relevant healthcare professionals.

OBJECTIVE To investigate the distribution,drug resistance and molecular characteristics of the Staph-ylococcus aureus strains isolated from intensive care unit(ICU)environment in some hospitals of Shanghai Pudong New Area so as to provide bases for prevention and control of infections.METHODS The specimens were collected from ICU environment of 8 medical institutions in Pudong District in 2019 and 2023,the S.aureus strains were i-solated and identified.The drug susceptibility testing was performed by microbroth dilution method.The drug re-sistance genes and virulence genes were predicted based on virulence factors of bacterial pathogens,the compre-hensive antibiotic resistance database(mcmaster.ca)and PubMLST,multilocus sequence typing(MLST)and phylogenetic analysis were performed.RESULTS Totally 3408 samples were collected,and the isolated S.aureus strains accounted for 1.29％(44/3408);the isolated strains form both the shared articles and the articles that health care workers directly contact accounted for 20.45％.The drug resistance rate to penicillin was highest(72.73％,32/44);the strains were completely susceptible to vancomycin,rifampicin,sulfamethoxazole-trime-thoprim,daptomycin and nitrofurantoin.The methicillin-resistant Staphylococcus aureus(MRSA)strains accoun-ted for 25.00％.The multi-drug resistance rate was 40.91％.The strains carried 26 types of drug resistance genes in total,covering 9 categories;the carrying rate of fluroquinolones resistance gene nor A was 100.00％.The strains carried 12 types of enterotoxin genes,the carrying rate of sea gene was highest(50.00％,22/44),some of the strains carried lukS-PV,lukF-PV and tsst gene.There were 14 types of sequence type(ST),dominated by ST7(13/44,29.55％),and novel ST type ST9342 was discovered.The phylogenetic analysis showed that all of the strains were divided into 2 major branches,and the strains with the same ST type were phylogenetically close on the evolutionary tree.CONCLUSIONS The S.aureus strains are prevalent in ICU environment and are highly re-sistant to antibiotics.The strains exhibit various ST types,which,to a certain degree,is associated with the path-ogenicity.

Objective:To compare the consistency and efficacy of ultrasound and CT in the preoperative diagnosis of cervical lymph node metastasis in patients with thyroid cancer, and to explore the clinical value of the combined application of multimodal imaging.Methods:The 119 thyroid cancer patients underwent surgical treatment from January to September 2023 in Zhongshan Hospital Affiliated to Dalian University were retrospectively analyzed. All patients underwent ultrasound and CT examinations before operation. The results of postoperative histopathology examination were taken as the gold standard, the efficacy of ultrasound and CT in preoperative diagnosis of cervical lymph node metastasis was compared.Results:A total of 1 721 cervical lymph nodes were detected in 119 patients with thyroid cancer, among which 1 378 lymph nodes were benign, and 343 lymph nodes were malignant, the rate of malignant lymph nodes was 19.93% (343/1 721). Among them, the proportion of malignant lymph nodes in area Ⅵ was the highest, 22.58% (245/1 085), followed by area Ⅲ, 21.26% (37/174). The sensitivity of CT in diagnosing cervical lymph node metastasis in patients with thyroid cancer was significantly higher than that of ultrasound diagnosis: 58.46% (38/65) vs. 38.46% (25/65), the specificity was significantly lower than that of ultrasound diagnosis: 85.19% (46/54) vs. 96.30% (52/54), and there were statistical differences ( P<0.01 and <0.05); there was no statistical difference in accuracy between CT and ultrasound ( P>0.05). Conclusions:In the preoperative diagnosis of cervical lymph node metastasis in patients with thyroid cancer by ultrasound and CT, ultrasound examination has no radiation risk, while CT examination has a higher diagnostic efficiency.

Objective:To explore the relationship between urinary cadmium levels and thyroid hormone levels in people aged 40-89 years old in selected areas of China.Methods:Based on the "Investigation of the Impact of Soil Quality of Agricultural Land on Human Health in Typical Areas" project from October 2019 to August 2020, a multi-stage stratified random sampling method was used to include 6 588 middle-aged and older adults aged 40-89. Demographic characteristics, dietary frequency and disease status were collected through the questionnaire and physical examination. Urinary cadmium and urinary creatinine were detected by random midstream urine. Fasting venous blood was collected for the detection of Triiodothyronine (T3) and Thyroxine (T4). The linear mixed effects model was used to explore the association of urine cadmium levels with thyroid hormone levels. Its dose-response relationship was explored by using the restricted cubic spline.Results:The age of the subjects was (63.48±12.18) years, with males accounting for 51.28%. The M ( Q 1,Q 3) of urinary cadmium level, T3 and T4 was 2.48 (1.36, 4.42) μg/g·creatinine, (1.96±0.51) nmol/L and (113.75±29.11) nmol/L, respectively. The linear mixed effects model showed that the changes of T3 and T4 were 0.027 (0.009, 0.044) nmol/L and 2.019 (1.084, 2.953) nmol/L for each one-unit increase (natural logarithm transformed) of urinary cadmium. The restricted cubic spline showed that there was a positive nonlinear association between urinary cadmium and T3 as well as T4 (all Pnonlinear<0.05). Conclusion:In selected areas of China, the urinary cadmium level of middle-aged and older adults aged 40-89 years is positively associated with T3 and T4.