OBJECTIVE To develop a method for the determination of tacrolimus （TAC） concentration in human whole blood and to apply it in clinical therapeutic drug monitoring. METHODS Whole blood samples were processed by protein precipitation with methanol. The determination was performed using liquid chromatography-tandem mass spectrometry （LC-MS/MS）， with ascomycin serving as the internal standard. Chromatographic separation was carried out on a Kinetex F5 100Å column with a mobile phase consisting of 0.1 mmol/L ammonium acetate containing 0.2 mmol/L formic acid and methanol. Gradient elution was performed at a flow rate of 0.4 mL/min. The injection volume was 5 μL. Detection was conducted using multiple reaction monitoring （ m / z 821.6→768.6 for TAC； m / z 809.4→756.1 for ascomycin） with an electrospray ionization source in positive ion mode. The study focused on 86 whole blood samples collected from 83 pedi atric patients who received TAC therapy at Children’s Hospital of Nanjing Medical University from September 1 to 30， 2025. The aforementioned method was employed to measure the TAC concentration in the whole blood samples. The correlation and agreement between the aforementioned method and the traditional enzyme multiplied immunoassay technique （EMIT） were evaluated through Spearman correlation analysis， Bland-Altman analysis， and Passing-Bablok regression analysis. RESULTS The linear range of TAC was 0.5-100 ng/mL； the evaluation results for accuracy， precision， extraction recovery， matrix effect， and stability tests all met the relevant requirements. Clinical application results showed that the median concentration of TAC in pediatric whole blood measured by LC-MS/MS and EMIT methods were 4.4 and 4.0 ng/mL， respectively. Moreover， the two methods exhibited a strong correlation （correlation coefficient of 0.848 1） and good agreement （average relative deviation of 6.5%）. CONCLUSIONS A reliable LC-MS/MS method for the determination of TAC concentration in human whole blood is successfully established. This method demonstrates strong correlation and good agreement with the EMIT method， making it suitable for clinical therapeutic drug monitoring.

Objective:To investigate the significance of urinary 8-oxo-7, 8-dihydroguanosine(8-oxoGuo)in assessing the short-term prognosis of sepsis in frail elderly patients.Methods:We conducted a cross-sectional study involving 62 frail elderly patients diagnosed with sepsis who were admitted to the Emergency Intensive Care Unit(EICU)at Beijing Hospital between March 2021 and March 2022.Based on their 28-day prognosis, the patients were categorized into two groups: those who died and those who survived.Upon admission, we collected urine samples and clinical data from both groups.We employed isotope dilution high-performance liquid chromatography-mass spectrometry to measure the levels of the RNA oxidation marker 8-oxoGuo in the urine.Results:A total of 62 frail elderly patients[aged(85.1±6.3)years]diagnosed with sepsis were included in the study, comprising 36 patients in the 28-day mortality group and 26 patients in the survival group.Univariate analysis revealed that the survival group had significantly lower body temperature, blood calcitonin(PCT)levels, sequential organ failure assessment(SOFA)scores, and urinary 8-oxoGuo levels compared to the mortality group.Additionally, the survival group exhibited a higher mean arterial pressure(MAP)than the mortality group, with all differences reaching statistical significance(all P<0.05).Spearman correlation analysis indicated that urinary 8-oxoGuo levels were positively correlated with both PCT and SOFA scores in frail elderly sepsis patients( r=0.426, 0.768, both P<0.05).Furthermore, logistic regression analysis identified urinary 8-oxoGuo and SOFA as independent risk factors for 28-day mortality in this population( OR=1.936, 1.427; P=0.006, 0.002).The area under the receiver operating characteristic curve(AUC)for urinary 8-oxoGuo and SOFA in predicting the 28-day prognosis of frail elderly sepsis patients was 0.761 and 0.741, respectively, both demonstrating statistical significance(both P<0.001). Conclusions:Our findings suggest that urinary 8-oxoGuo possesses strong predictive value for the short-term prognosis of sepsis in this vulnerable population.

Purpose This study aimed to investigate the expression of CTLA-4 and PD-L1 in pulmonary lympho-epithelial carcinoma(PLEC)and to explore their relationships with patient prognosis and with tumor-infiltrating lym-phocytes(TILs).Methods Fifty cases of PLEC were retrospectively collected,together with 23 samples of adjacent normal lung tissue.Immunohistochemistry was performed to detect CTLA-4 and PD-L1 expression in both PLEC and adjacent normal lung tissues,as well as to quantify CD4+and CD8+T-lymphocytes infiltration within the tumor micro-environment.CTLA-4,PD-L1,and the distributions of CD4+T cells and CD8+T cells were then correlated with the clinicopathological features of PLEC.Results The positive rate of CTLA-4 in PLEC was significantly higher than that in adjacent normal lung tissue(P＜0.05).PD-L1 expression differed significantly across TNM stages of PLEC(P＜0.05)and was positively correlated with TNM stages(r=0.31,P=0.03).CD4+and CD8+T-lymphocytes were pre-dominantly localized in the tumor stroma,with CD4+T cells density exceeding that of CD8+(P＜0.05).Within canc-er nests,CD8+T cells density was significantly higher than CD4+(P＜0.05).Conclusion Both PD-L1 and CTLA-4 are frequently expressed in PLEC,suggesting they represent potential immunotherapeutic targets.In the PLEC micro-environment,lymphocytes primarily infiltrated the stromal compartment,and CD4+T cells are more abundant than CD8+T cells in that locale.

The neurovascular unit (NVU) plays a crucial role in the functionality and plasticity of the visual cortex by tightly coupling neuronal activity with vascular function through the coordinated action of multiple cell types.Synaptic activity in neurons drives glutamate uptake by astrocytes, triggering calcium signaling, which further modulates local blood flow dynamics to meet the metabolic demands of the visual cortex during periods of high neural activity.Neurons also contribute to synaptic plasticity through mechanisms such as long-term potentiation and long-term depression, determining the efficiency of visual information processing and network remodeling.Astrocytic calcium signaling serves as a vital intermediary in this process, not only supporting the metabolic needs of neurons but also shaping the fine structure of visual networks by regulating synaptic dynamics.During critical periods, astrocytic activity within the NVU promotes synaptic maturation and establishes the temporal window for plasticity in the visual cortex.In adulthood, the proper functioning of NVU provides a foundation for the reorganization of visual function.Dysfunctional NVU activity can impair visual plasticity, highlighting its relevance as a therapeutic target for conditions such as amblyopia, brain injuries, and neurodegenerative diseases.Understanding the collaborative mechanisms between neurons and other NVU components offers novel theoretical insights and practical approaches for restoring visual function and intervening in neurological disorders.This article reviews the research progress of NVU in visual plasticity, focusing on the concept of NVU, the roles of neurons and the functions of astrocytes within NVU, as well as the roles of NVU in synaptic plasticity, critical period visual plasticity, and plastic remodeling.

Objective To explore the mechanism of Tongdu Xingshen acupuncture,the clinical efficacy,systemic inflammatory response,blood-brain barrier and intestinal flora in patients with cognitive impairment after ischemic stroke(IS)were studied.Methods Thirty patients(3 cases shedding)with cognitive impairment after IS were included as the disease group,including patients before treatment as the disease group,patients after Tongdu Xingshen acupuncture treatment as the electroacupuncture group,and 30 healthy controls(3 cases shedding)were included as the healthy group.In the electroacupuncture group,on the basis of the basic treatment,Tongdu Xingshen acupuncture was applied,which was 30 min each time,once a day for 14 days.The MMSE,MoCA and MBI scores of the three groups were observed.The fecal and serum samples from all study subjects were collected,and 16S rDNA sequencing technology and ELISA were used to detect the changes of proinflammatory factors IL-6,IL-1β,TNF-α and S100β in serum in intestinal flora and feces.Results Compared with the healthy group,the MMSE,MoCA,and MBI score of patients in the disease group decreased significantly(P<0.05),serum proinflammatory factors and S100β protein content increased significantly(P<0.05),and the Shannon index(P<0.01)and Simpson index(P<0.001)increased significantly.Compared with the disease group,the MMSE,MoCA,and MBI score of the EA group increased significantly(P<0.05),the serum levels of proinflammatory factors and S100β decreased significantly(P<0.05),Shannon index and Simpson index decreased(P>0.05).The dominant bacterial flora in the healthy group mainly included Bacteroides,Bifidobacterium,Bacteroides,Faecalibacterium,Bifidobacteriaceae,Ruminococcaceae,and Bacteroides and other beneficial bacteria(P<0.05).The dominant flora in the disease group included Proteobacteria,Enterobacteriaceae,Escherichia,Klebsiella and other opportunistic bacteria(P<0.05),while the dominant flora in the EA group was consistent with the healthy group,the relative abundance of beneficial bacteria increased significantly(P<0.05),and the relative abundance of opportunistic bacteria decreased significantly(P<0.05).Spearman correlation analysis found that beneficial bacteria were positively correlated with clinical efficacy related indicators,but with serum proinflammatory factors and the content of S100β was negatively correlated.Conclusion Tongdu Xingshen acupuncture can regulate the diversity of intestinal flora to increase the abundance of Bacteroides,Bifidobacterium,Faecalibacterium,and other beneficial bacteria,regulate the intestinal microecological balance,Thereby regulating systemic inflammation and blood-brain barrier function,which plays a role in improving cognitive function.

Objective:To understand the experience of inpatients′ needs on medical nursing assistants, and explore the characteristics and population differences, in order to provide a reference for optimizing the inpatient companion management system.Methods:The descriptive qualitative study was adopted for semi-structured in-depth interviews of 19 inpatients hospitalized in Zhongshan Hospital Fudan University from December 2023 to January 2024 based on purposive sampling. Information on personal characteristics, status and experience of dependence on medical nursing assistants was collected. Colaizzi seven-step analysis method was used to construct the fact labels and model dimensions. Manually extract the characteristics of inpatients′ needs on medical nursing assistants, and construct a user persona to complete the visualization.Results:A total of 19 inpatients′ were interviewed, including 12 males and 7 females with an average age of (63.16 ± 11.56) years. Four dimensions were extracted including cognitive characteristics, health status characteristics, family support characteristics, and psychological characteristics. Combined with personal traits and experience, three types of typical user persona were constructed including independent type, mutual assistance and cooperation type, complete dependence type.Conclusions:Group heterogeneity existed in the demand of inpatients on medical nursing assistants. According to the user persona, the manager should formulate personalized and precise caregiver management strategies.

Objective To investigate the acceptance and satisfaction of tuberculosis prevention and control personnel in Beijing with the patient management system, and to provide a basis for further improving the patient management model. Methods A survey was conducted on the current usage, satisfaction, willingness to use and system improvement opinions of the patient management system among medical staff involved in the supervision and medication management of pulmonary tuberculosis patients in Beijing. Results A total of 360 medical staff participated in the survey. ^“Patient management^” was the function with the largest number of users, accounting for 96.94%. The proportion of users of each module who believed that the module's design met actual work needs was over 90%. About 94.44% of respondents believed that patient management systems facilitated the transfer and sharing of information between institutions. And 90.83% of respondents thought that the patient management system was easy to operate, and 89.17% of respondents believed that patient management systems reduced workload. About 97.50% of respondents were satisfied with the overall use of the patient management system. The results of the influencing factor analysis showed that those with 3 or less modules designed to meet actual work were less satisfied than those with more than 3 modules, and the difference was statistically significant (P=0.001). Respondents put forward suggestions for improvement on the optimization of operational details such as system response speed, interface design, system login and query statistics. Conclusion Medical staff involved in the follow-up management of pulmonary tuberculosis patients are highly satisfied with their work using the patient management system. During the promotion and use, it is still necessary to continuously optimize the system functions according to work needs so that the system can truly facilitate work.

Post-cholecystectomy syndrome (PCS) encompasses persistent or new abdominal pain, bloating, and diarrhea following cholecystectomy. Our understanding of its etiology, diagnosis, and treatment has evolved significantly. This systematic review traced the conceptual progression of PCS and addressed clinical controversies, and reflections on diagnostic and therapeutic improvements. The definition of PCS has shifted from an anatomical focus (e.g., retained stones, biliary duct injury) to functional disorders (e.g., sphincter of Oddi dysfunction, abnormal bile acid metabolism, and psychosomatic factors). Current diagnosis strictly adheres to the Rome Ⅳ criteria, with an approximate prevalence of 10%. Historically broad diagnostic criteria explained the wide variability in reported incidence rates (5%-63%). Ambiguity persists regarding whether pre-existing symptoms persisting or evolving postoperatively should be attributed to PCS.Therapeutic approaches have transitioned from definitive surgical interventions for organic lesions to pharmacological management of functional dyspepsia. Given the inherent conceptual ambiguity in PCS, we proposed replacing PCS with ​post-cholecystectomy biliary dyspepsia (PCBD)—a term emphasizing its postoperative onset, functional dyspepsia characteristics, and exclusion of preoperative symptoms or non-biliary etiologies. The introduction of the concept of PCBD can help to unify diagnostic criteria, guide individualized treatment, and conduct in-depth research.

Objective To retrospectively analyze multicenter data from domestic sources, aiming to explore the link between intrahepatic cholangiocarcinoma (ICC) tumor size and prognosis, establishing a classification system based on tumor size. Methods Between December 2011 and September 2018, 280 ICC patients from 13 hospitals were included. The tumor size prognosis cutoff was identified by the minimum P-value method, and the classification's overall survival related effectiveness was assessed by Kaplan-Meier analysis. Results All 280 patients were divided into the group of tumor maximum diameter ≤4 cm and >4 cm. Tumor size was confirmed as an independent prognosis factor by multivariate COX regression analysis (HR=2.110, 95% CI: 1.358-3.280). Conclusions The tumor size dichotomy classification system based on the Chinese patient group can expediently predict ICC prognosis and offers an important basis for selecting post-operative individualized adjuvant therapy and follow up plans.

Objective To analyze the potential profiles of discharge readiness in patients after 125I seed implantation and the influencing factors of different profiles.Methods A total of 221 patients who underwent 125I seed implantation in the Interventional Therapy Department,Zhejiang Cancer Hospital from October 2023 to October 2024 were selected as the research objects by convenient sampling method.The general information questionnaire,discharge readiness scale and social support rating scale were used for investigation.Mplus8.3 software was used to conduct latent profile analysis on the discharge readiness of patients after 125I seed implantation,and multivariate Logistic regression model was used to analyze the influencing factors of different discharge readiness profiles.Results The discharge readiness of patients after 125I seed implantation was divided into three potential profiles,"low discharge readiness-low self-care group"(10.41％),"medium discharge readiness-low others care group"(32.58％)and"high discharge readiness-high emotional support group"(57.01％).Logistic regression analysis showed that the number of short-term complications and the level of social support were the influencing factors of the potential profiles of discharge readiness in patients after 125I seed implantation(P＜0.05).Conclusion There is heterogeneity in the discharge readiness of patients after 125I seed implantation,and personalized intervention strategies should be formulated according to different profiles.