OBJECTIVE To establish a method for simultaneous determination of plasma concentration of lamotrigine（LTG）， levetiracetam（LEV） and perampanel（PER） in children with epilepsy and apply this method in clinical practice. METHODS Plasma proteins were precipitated with acetonitrile. Using PER-D 5 as internal standard， ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method was adopted. The determination was performed on ACQUITY UPLC HSS T3 C 18 column with mobile phase consisted of 0.1% formic acid with 5 mmol/L ammonium acetate-acetonitrile （gradient elution） at the flow rate of 0.3 mL/min. The column temperature was 40 ℃， and sample size was 5 μL. The analysis time was 5 min. The electrospray ionization source and multiple reaction monitoring mode were used for positive ion scanning. The ion pairs used for quantitative analysis of LTG， LEV， PER and internal standard were m / z 255.9→144.9， m / z 171.1→126.1， m / z 350.1→219.0 and m / z 354.9→220.2， respectively. The steady-state trough concentrations of the aforementioned drugs in the plasma of 14 pediatric epilepsy patients receiving combination therapy were determined using the same UPLC-MS/MS method as above. RESULTS The linear ranges of LTG， LEV and PER were 0.15-24 μg/mL （ R 2 ＞0.993）， 0.312 5-50 μg/mL （ R 2 ＞0.997） and 6.25-1 000 ng/mL （ R 2 ＞0.997）， respectively. The lower limits of quantification were 0.15 μg/mL， 0.312 5 μg/mL and 6.25 ng/mL， respectively. RSDs of intraday and interday precision tests of the three drugs were no more than 9.83%， and the accuracies （relative errors） were between －9.33% and 13.72%（ n ＝6 or n ＝18）； the average extraction recovery rates were 86.4%-97.9%， and the average matrix effects were 86.9%-110.0% （ n ＝6）. The absolute values of the relative errors in the stability tests were all below 15%. The steady-state trough concentrations of LTG， LEV and PER were （5.64±4.03）μg/mL， （10.67±8.78）μg/mL and（450.20±251.27）ng/mL， respectively； the rates of achieving target trough concentrations were 71.4%， 37.5% and 84.6%， respectively. CONCLUSIONS The established UPLC-MS/MS method is specific， rapid and suitable for the plasma concentration monitoring in epileptic children receiving combination therapy.

Objective To investigate the risk factors for ICU-acquired weakness (ICU-AW) in patients with sepsis, and to construct a nomogram prediction model. Methods A total of 360 patients with sepsis admitted from January 2020 to December 2025 were enrolled as study subjects. The risk factors for ICU-AW were identified by logistic regression analysis. A nomogram prediction model was constructed, and the model was validated. Results Multivariate logistic regression analysis indicated that age, APACHE II score, duration of mechanical ventilation, duration of immobilization, duration of sedation, and NLR were risk factors for the occurrence of ICU-AW (P<0.05), while ALB was a protective factor (P<0.05). A nomogram model was constructed based on these seven variables. After validation via Bootstrap resampling, ROC curve showed an AUC of 0.949 (0.929–0.969) and a corrected AUC of 0.942 (0.940–0.948), indicating good discrimination degree of the model. Hosmer-Lemeshow test for calibration curve revealed a chi-square value of 10.188 (P=0.252), suggesting that the predicted value of fitting was generally consistent with the actual value. Decision curve analysis demonstrated that when the threshold was in the range of 0.05–0.9, the model had a good net benefit. Conclusion The nomogram prediction model for sepsis ICU-AW based on seven indicators including age, APACHE II score, NLR, ALB, and the durations of mechanical ventilation, immobilization, and sedation exhibits good predictive efficiency and stability. This nomogram prediction model is helpful for the clinical rapid screening of ICU-AW high-risk groups in patients with sepsis, and meets clinical needs for early warning and precise prevention and control.

Objective To investigate the protective effects of Wuling capsule on mice with autoimmune hepatitis(AIH).Methods Mice were randomly divided into control group,AIH model group,Wuling capsule low-dose group(0.5 g·kg-1·d-1),Wuling capsule middle-dose group(1.0 g·kg-1·d-1),and Wuling capsule high-dose group(2.0 g·kg-1·d-1),with 10 mice in each group.The Wuling capsule groups were administered with Wuling capsule suspension of different doses orally at a volume of 10 mL/kg once daily;the control group and AIH model group were given the same volume of saline by gavage.After 14 d of administration,mice in the AIH model group and Wuling capsule groups were injected with concanavalin A(20 mg/kg)via the tail vein,and the serum,liver,and spleen were collected 8 h after injection.Serum alanine transaminase(ALT)and aspartate transaminase(AST)levels were measured using an automatic biochemical analyzer;hematoxylin-eosin staining was used to observe the pathological structure of the liver tissue;the contents of interleukin(IL)-6,IL-4,and tumor necrosis factor(TNF)-α in liver were determined by enzyme-linked immunosorbent assay,and quantitative polymerase chain reaction was used to determine the relative mRNA expression levels of IL-6,IL-4,TNF-α,Toll-like receptor 4(TLR4),and nuclear factor κB(NF-κB)in the liver.Fluorescence immunoassay was used to analyze the expression and nuclear translocation of NF-κB p65 protein in the liver.Results Compared with the control group,the AIH model group showed abnormal liver morphology and structure,increased serum ALT and AST levels,increased contents of IL-4,IL-6,and TNF-α in the liver,upregulated mRNA expression levels of IL-6,IL-4,TNF-α,TLR4,and NF-κB in the liver,and increased nuclear entry of NF-κB p65.Wuling capsule significantly improved the pathological structure of the liver in AIH mice,reduced serum ALT and AST levels,decreased the contents of IL-4,IL-6,and TNF-α in the liver and the mRNA expression levels of TLR4,NF-κB,IL-4,IL-6,and inhibited the nuclear activation of NF-κB p65.Conclusion Wuling capsule has significant protective effects on AIH mice,which may be related to the TLR4/NF-κB signaling pathway.

Objective To explore the psychological feelings and coping resources of patients waiting for a liver transplantation,in order to provide references for implementing psychological nursing care.Methods Using purposive sampling,15 patients waiting liver transplantation surgery from June to November 2024 at a tertiary hospital in Qingdao were selected as study subjects.Semi-structured in-depth interviews were conducted,and data were analyzed using interpretative phenomenological analysis to extract themes.Results The psychological feelings of patients waiting for a liver transplantation included 2 main themes with 6 sub-themes,namely,the complex and contradictory psychological feelings of liver transplant patients during the waiting period(expectations for transplanta-tion and future outlook,multiple psychological burdens intertwining,introspection and growth),and mobilizing psychological resilience resources to deal with complex psychological feelings(core resources,internal resources,external resources).Conclusion Clinical medical staff should implement psychological care for patients waiting for a liver transplantation based on the complex and contradictory psychological feelings they experience.Also,they should target the coping resources for psychological feelings to construct coping strategies,thereby comprehensively improving patients' psychological health during the waiting period.The coping strategies include promoting patients' health behaviors,improving their psychological status,and providing multidimensional social support for them.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective:To develop a deep learning-based segmentation model for anterior segment ultrasound biomicroscopy (UBM) images to automatically segment the anterior segment tissues of patients with primary angle-closure glaucoma (PACG).Methods:A single-center retrospective case series was conducted.A small-scale dataset comprised 468 UBM images of the anterior chamber angle closure from 156 patients with PACG who underwent the UBM examination at Tianjin Medical University Eye Hospital between July 12, 2022, and February 20, 2023.The UBM images were randomly split into a training dataset of 228 images and a testing dataset of 152 images using a random seed method in a ratio of 6∶4.The models were trained using the PSPNet model with MobileNet V2 and ResNet50 as backbones, the DeepLab v3+ model with MobileNet V2 and Xception as backbones, and the SegFormer model with MiT-B0 and MiT-B2 as backbones.The testing dataset was used for result prediction and to achieve segmentation of four regions: the cornea and sclera, iris, ciliary body, and anterior lens surface.To evaluate the performance of the models in segmenting the anterior segment structures, multiple metrics were assessed, including the mean intersection over union (mIoU), Dice coefficient, precision, recall, false negative rate, and specificity.A comparative analysis of the test results across the different models was subsequently performed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Tianjin Medical University Eye Hospital (No.2023KY-05).Results:The two models with the best segmentation performance were PSPNet and DeepLab v3+ .The PSPNet model with ResNet50 as the backbone achieved the mIoU of 85.11%, Dice coefficient of 91.38%, precision of 91.83%, recall of 90.94%, false negative rate of 9.06%, and specificity of 98.89%.The DeepLab v3+ model with MobileNet V2 as the backbone achieved an mIoU of 85.84%, Dice coefficient of 92.01%, precision of 92.67%, recall of 91.36%, false negative rate of 8.64%, and specificity of 98.90%.Among the five key metrics, mIoU, Dice coefficient, recall, false negative rate, and specificity, DeepLab v3+ exhibited the best segmentation performance.In addition, the DeepLab v3+ model with Xception as the backbone had the highest precision among all models, reaching 92.77%.Conclusions:The deep learning-based DeepLab v3+ model achieves precise segmentation of anterior segment tissue structures in PACG anterior segment UBM image segmentation, providing auxiliary support for clinical diagnosis.

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

Aim To investigate the effects of the fangchinoline derivative LYY-32 on the biological prop-erties of the BLM642-1290 DNA helicase,in order to lay a foundation for further research on its antitumor activity.Methods Fluorescence polarization assay,malachite green-phosphate and ammonium molybdate colorime-try,and fluorescein-labeled DNA gel electrophoresis experiments were conducted to study the effects of fangchinoline derivative LYY-32 on the DNA binding activity,ATPase activity,and DNA unwinding activity of BLM642-1290 DNA helicase.The effects of LYY-32 on the DNA unwinding activity of DNA helicase in cells were studied using fluorescent techniques and time-lapse microscopy.Ultraviolet spectral scanning was used to investigate the effects of LYY-32 on the confor-mation of the BLM642-1290 DNA helicase.Results At a concentration of 10 μmol·L-1,the inhibition rate of LYY-32 on BLM642-1290 DNA helicase binding to dsDNA was 53.17％.At a concentration of 5 μmol·L-1,the inhibition rate of LYY-32 on BLM642-1290 DNA helicase binding to ssDNA was 88.49％.The inhibition rate of LYY-32 on the ATPase activity of BLM642-1290 DNA he-licase was 89.3％at a concentration of 50 μmol·L-1.When the concentration of LYY-32 exceeded 5μmol·L-1,its inhibition rate on the DNA unwinding activity of BLM642-1290 DNA helicase was 100％.LYY-32 also significantly inhibited the DNA unwinding ac-tivity of DNA helicase in cells.However,LYY-32 had no effect on the conformation of BLM642-1290 DNA heli-case.Conclusion The DNA binding activity,AT-Pase activity,and DNA unwinding activity of BLM642-1290 DNA helicase could be significantly inhibi-ted by the fangchinoline derivative LYY-32.

This study was aimed at understanding the prevalence and influencing factors of food-borne parasitic diseases in ethnic minority areas of Guizhou Province,to provide a scientific basis for the development of appropriate intervention measures based on the human-animal-environment One Health concept.In 2023,the infection status of the human population,reservoir hosts,intermediate hosts,food-borne parasitic diseases,and related social and environmental factors were investigated in Congjiang County in Qidongnan Miao and Dong Autonomous Prefecture;Luodian County in Qiannan Buyi and Miao Autonomous Prefecture;and Ceheng County in Qianxinan Buyi and Miao Autonomous Prefecture.At least 1 000 individuals were sampled from each county,along with at least 50 insect-protected host samples from each location.Food-borne parasite infections were detected with the modified Kato thick smear method.A questionnaire survey was administered to the population.Detection of food-borne parasitic metacercariae was performed in intermediate host fish through the flaking and digestion method,and in crabs through the pounding and sedimentation method.The chi-square test was used to compare rates,and logistic regression was applied for multivariate analysis.A total of 3 023 questionnaires and fecal samples were collected.Males accounted for 47.50%,females accounted for 52.50%,and members of ethnic minorities accounted for 96.06%.A total of 186 food-borne parasitic infections were identified,and the infection rate was 6.15%.Five insect species were detected,which showed an infection rate of 5.39%.The infection rate of Clonorchis sinensis was 0.33%,that of Taenia was 0.40%,that of Heteroceles was 0.17%,that of Acanthus was 0.17%,and that of Echinostoma was 0.03%.Human infections with Echinostomus colloides and Echinostomia transferoris had not previously been reported in China.Single-factor analysis revealed statistically significant differences in food-borne parasite infections according to various factors,including the consumption of untreated water,raw fish and shrimp,raw pig blood,raw cow gastric juice,and raw pork and beef,as well as raw pig and cow viscera(P<0.05).Multivariate analysis indicated that the risk factors for food-borne parasite infections among residents in minority areas of Guizhou Province included the consumption of raw pig blood(OR=2.841,95%CI:1.809-4.463),raw cow gastric juice(OR=2.122,95%CI:1.297-3.469),and raw fish and shrimp(OR=1.779,95%CI:1.049-3.018).A total of 173 fecal samples of the reservoir host were examined,which showed a rate of food-borne parasite infection of 5.2%.A total of 510 intermediate host fish were examined,which showed a 4.51%positivity rate of encysted metacercaria of Clonorchis sinensis.The crab,pig,and beef samples were not positive.In conclusion,food-borne parasitic infections were prevalent in ethnic minority regions of Guizhou Province,and consumption of raw food were influencing factors.A focus on populations with raw food consumption habits,including raw pig blood,cow gastric juice,fish and shrimp,is essential.Concurrently,monitoring of animal hosts must be strengthened to perform key interventions according to the One Health concept.

Chrysophanol(Chr)and physcion(Phy)are primary active ingredients of a well-known traditional Chinese medicine namely rhubarb(Chinese name:Dahuang),and their glucuronides have been revealed as the dominant forms presenting in rats after oral administration.Either Chr or Phy has two glycosylation sites,resulting in a pair of positional isomers for glucuronides of either compound(CG1&CG2 and PG1&PG2).To confirmatively identify these glucuronides,energy-resolved mass spectrometry(ER-MS)was used to pursue the fragmentation trajectories of the targeted fragment ions,and the resultant breakdown graphs that were described by the optimal collision energy(OCE)were expected to exhibit the differences of glycosidic bond cleavage between the isomers.Quantum chemical calculation was thereafter conducted to produce the bond dissociation energy(BDE)of the glycosidic bonds.The isomers were unambiguously identified through applying the positive correlation rule between OCE and BDE.Fortunately,the glucuronides of Chr and Phy in vivo were observed through liver microsomes incubationin vitro.ER-MS was utilized to collect the Gaussian-shaped breakdown graphs in response to the neutral loss of 176 Da,and the absolute values of OCE were compared between positional isomers.The results revealed that CG1(-32.31 eV)>CG2(-31.61 eV),and nonetheless,PG1(-30.00 eV)Chr-8-GluA(-48.787 kJ/mol),and nonetheless,Phy-1-GluA(-48.672 kJ/mol)