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 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.

Objective To analyze the effects of tyrosinase on choroidal and retinal thickness and blood flow changes in guinea pigs,and to explore the role of tyrosinase in the development and progression of myopia.Methods A total of 30 three-week-old male tri-colored guinea pigs and 10 albino guinea pigs were selected and divided into four groups:control group(tri-colored hyperopic guinea pigs,with no intervention),albino group(albino myopic guinea pigs,with no inter-vention),myopia group(tri-colored myopic guinea pigs,with no intervention),and injection group[tri-colored hyperopic guinea pigs,injected with tyrosinase inhibitor(6 250 μg·L-1),100 μL per day].The experiment lasted for 4 weeks.The refractive status and axial length(AL)of the guinea pigs in each group were measured,along with ocular biometric param-eters related to axial length[anterior chamber depth,aqueous humor depth(AQD),central corneal thickness,lens diame-ter,vitreous chamber depth(VCD)].Optical coherence tomography was used to measure the retinal thickness,choroidal thickness(ChT),and choroidal blood perfusion(ChBP)in different regions of the guinea pigs.The tyrosinase activity in the vitreous and retina of guinea pigs in each group was measured.The expression levels of neurotransmitters in the vitre-ous and retina of guinea pigs in each group were detected.Results The differences in refractive status between the albi-no group and the control group at 0,2,and 4 weeks of the experiment were statistically significant(F=8.972,P＜0.05).At the 4th week of the experiment,the refractive status of the injection group was lower than that of the control group,and the AL was greater than that of the control group,with statistically significant differences(both P＜0.05).Further analysis of AL-related biometric parameters revealed that only AQD and VCD were associated with the changes in AL of guinea pigs in each group.At the 0th week of the experiment,the average retinal thickness of the control group was greater than that of the albino group,with a statistically significant difference(t=-9.007,P＜0.000 1).Moreover,the differences in reti-nal thickness in the outer retina across different directions and time points were statistically significant(all P＜0.05).The differences in retinal thickness between the control and albino groups were mainly concentrated in the outer retina.The ChT of the albino group was less than that of the control group,with a significant difference between groups(F=4.809,P=0.030).The ChBP of the control group was significantly higher than that of the albino group at 0,2,and 4 weeks of the experiment(all P＜0.05).At the 4th week of the experiment,the tyrosinase activity in the vitreous of the injection,albi-no,and myopia groups was lower than that of the control group,with statistically significant differences(all P＜0.05).The differences in neurotransmitters between the albino and control groups were mainly concentrated in the vitreous.In the retina,the ornithine level in the myopia group was higher than that in the albino and control groups,and the tryptophan levels in the myopia and control groups were higher than that in the albino group,with statistically significant differences(all P＜0.05).Conclusion Tyrosinase plays a crucial role in the development of myopia,regulating the development of refractive status by influencing the physiological properties of the retina and choroid.

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.

Objective:To evaluate the outcomes of intensive conservative treatment compared to conventional conservative treatment in patients with acute aortic syndrome(AAS).Methods:The study prospectively enrolled consecutive patients with AAS who were admitted to Beijing Anzhen Hospital, affiliated with Capital Medical University, and Beijing Dawanglu Emergency Rescue Hospital from January 2024 to December 2024. These patients with surgical contraindications or refused surgery for various reasons opted for conservative treatment. A total of 282 patients were included, and 15 patients with missing data or those who died without any treatment were excluded. Finally, 267 patients were enrolled, of whom 94 received intensive conservative treatment, and 173 received conventional conservative treatment, the inverse probability of treatment weighting (IPTW) was used to reduce the influence of confoundings. After adjusting of baseline datas via IPTW, the survival outcomes of the two groups were compared at 14 days, 30 days, and at the end of follow-up.Results:The results showed significant differences in acute phase survival rates between the enhanced conservative treatment group and the conventional conservative treatment group at 14 days(82.40%vs.53.20%, P<0.0001). Significant survival differences were also observed at 30 days and at 276-day mid-term follow-up (96.29% vs.51.60%, P<0.0001; 78.50% vs.48.50%, P<0.0001). In the subgroup analysis, for type A aortic dissection, the enhanced conservative treatment group had higher survival rates compared to the conventional conservative treatment group at 14, 30 and 276 days (63.46% vs.41.35%, P<0.05; 52.17% vs.37.90%, P<0.05; 50.00% vs. 31.97%, P<0.05). However, for type B aortic dissection, although the enhanced conservative treatment group had higher survival rates than the conventional conservative treatment group, no statistically significant differences were observed (96.29% vs. 80.00%, P=0.054; 95.65% vs.78.37%, P=0.067; 94.12% vs.74.20%, P=0.088). Conclusion:For patients diagnosed with AAS are forced to choose conservative treatment if emergency surgery is not possible in the first place, intensive conservative treatment strategies can significantly reduce the mortality in the acute phase compared with conventional conservative treatment. Mid-term follow-up, intensive conservative treatment still has a significant survival advantage.

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 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.