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:Analyze the clinical features in children with pharyngeal neoplasm, and explore the clinical efficacy of low-temperature plasma minimally invasive treatment in children with pharyngeal neoplasm. Methods:A total of 46 pediatric cases of pharyngeal neoplasms admitted to the Otolaryngology Department of Wuhan Children's Hospital from January 2016 to December 2023 were included. There were 23 males and 23 females, with admission ages ranging from 5 days to 9 years and 4 months, and a median age of 2 years and 2 months. The clinical manifestations, examinations, treatments, and outcomes were evaluated. Results:Among the 46 pediatric patients, 21 were admitted with throat wheezing, 7 with masses in the oropharynx or head and neck, 5 with dyspnea, 4 with snoring during sleep as the main symptom, 3 with hoarseness, 2 with sore throat, 2 with swallowing discomfort, and 2 with difficulty feeding as the first symptom. Comorbidities included 5 cases of combined laryngomalacia, 4 of snoring, 4 of congenital heart disease, 4 of severe pneumonia, 3 of myocardial damage, 1 of multiple deformities, and 1 after hematopoietic stem cell transplantation. All patients underwent CT or MRI examination, with 26 cases undergoing local ultrasound examination and 39 undergoing electronic laryngoscopy examination. A total of 19 patients were transferred to the ICU after surgery, all of whom were patients with pharyngeal cysts. All patients underwent low-temperature plasma minimally invasive surgery, and all patients were diagnosed through pathological examination, including 1 case of nasopharyngeal teratoma, 5 of pharyngeal non-Hodgkin's lymphoma, 1 of oropharyngeal rhabdomyosarcoma, 1 of laryngeal perivascular epithelioid cell tumor （PEComa）, and 38 of pharyngeal cysts （2 cases of uvula, 2 of oropharynx, 7 of epiglottic valley, 19 of tongue base, 2 of branchial cleft cyst, and 6 of throat）. Conclusion:Pharyngeal neoplasms in children are prone to upper airway obstruction, including some rare or malignant tumors. Imaging and pathological diagnosis can assist in early diagnosis. Depending on the nature of the tumor, early detection and treatment can improve the quality of life and survival rate of children. Low-temperature plasma radiofrequency ablation is safe, minimally invasive, precise, and provides a clear field of vision, making it a valuable treatment option for children with pharyngeal neoplasms.
Humans ; Male ; Female ; Child, Preschool ; Child ; Infant ; Pharyngeal Neoplasms/therapy* ; Minimally Invasive Surgical Procedures/methods* ; Cold Temperature ; Treatment Outcome

Objective:To establish a patient-derived tumor xenograft (PDX) model of primary liver cancer (PLC), and to analyze the pathological features, proliferation and drug-related gene mutation characteristics of the primary tumor and the PDX model of primary liver cancer.Methods:A retrospective analysis was conducted on the tumor samples of 64 PLC patients who underwent hepatectomy in the Department of Hepatobiliary Surgery of Tianjin First Central Hospital from January 2019 to December 2020. Among them, there were 46 males and 18 females, with an average age of (60.7±9.4) years. The degree of tumor differentiation and whether there was vascular invasion were recorded. The pathology of the primary tumor and each generation of PDX models was observed. Immunohistochemical staining was used to detect the characteristic proteins and proliferation-related protein of PLC. The genes related to drug use in PLC PDX models were analyzed in the primary tumor.Results:Among the tumor tissues of 64 PLC patients, 31 cases (48.4%) were successfully transplanted into nude mice and passaged to subsequent generations. In the primary tumor tissues and PDX models of hepatocellular carcinoma (HCC), the cancer nodules were clearly distinguishable, the cancer cells were arranged disorderly, and distributed in nests or cords. The tumor cell nuclei were large and deeply stained. In the primary tumor tissues and each generation of PDX models of intrahepatic cholangiocarcinoma (ICC), there were ICC adenocarcinoma-like structures, with well-differentiated tumor glands and glandular structures composed of cuboidal or columnar tumor cells, presenting as small individual glands or interwoven glands. The degree of differentiation of the PDX models of HCC and ICC patients was basically consistent with that of the primary tumors. Immunohistochemistry showed that proliferating cell nuclear antigen staining of the primary tumors and PDX model transplanted tumors of HCC and ICC patients was strongly positive. The Ki-67 staining positive rate of the primary tumors and PDX model transplanted tumors of HCC was > 80%, and that of ICC was > 70%. Alpha-fetoprotein was strongly positive in the primary tumors and PDX model transplanted tumors of HCC and ICC. The common mutations of transplanted tumors and the primary tumors of P24 HCC patients were 90%, and those of P43 ICC patients were 89%, 94%, and 94%, respectively.Conclusion:The constructed PDX model is highly consistent with the biological characteristics of the primary tumor.

Objective:To explore the molecular mechanism by which interferon regulatory factor 1 (IRF1) affects hepatic ischemia-reperfusion injury (HIRI) by regulating the polarization of Kupffer cells.Methods:Twelve male healthy C57BL/6 wild-type mice weighing 20-25 g and aged 6-8 weeks were divided into a sham operation group ( n=6) and a HIRI group ( n=6); Twelve male healthy C57BL/6 IRF1 gene knockout (IRF1 -/-) mice weighing 20-25 g and aged 6-8 weeks were divided into a sham operation IRF1 -/- group ( n=6) and a HIRI IRF1 -/- group ( n=6). The levels of serum alanine transaminase (ALT) and aspartate transaminase (AST) in mice were measured, and hematoxylin-eosin (HE) staining of liver tissues was performed for Suzuki scoring to evaluate liver injury. Fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to evaluate the mRNA levels of IRF1 and tumor necrosis factor α (TNFα) in liver tissues. Flow cytometry and qRT-PCR were used to detect the proportion and functional changes of M1/M2-type Kupffer cells in liver tissues. IRF1 was overexpressed or knocked down in the mononuclear macrophage cell line ANA1, and a co-culture and hypoxia-reoxygenation system with the hepatocyte cell line AML12 was established. Flow cytometry was used to detect the apoptosis of AML12 cells. Results:At 12 hours after hepatic ischemia-reperfusion in wild-type mice, the liver tissue injury was the most severe. Compared with the sham operation group, the levels of serum ALT [(8 073±83) U/L vs. (81±19) U/L, q=13.59] and AST [(11 170±2 890) U/L vs. (412±210) U/L, q=13.77] in the HIRI group were significantly higher, and the differences were statistically significant (both P<0.001). The Suzuki score reached 5-6 points. At 12 hours after hepatic ischemia-reperfusion in IRF1 gene knockout mice, the liver tissue injury was not obvious. There were no significant differences in the levels of serum ALT [668 (514, 2 344) U/L vs. 254 (147, 285) U/L, q=2.52, P=0.348] and AST [1 936 (1 262, 2 003) U/L vs. 628 (423, 759) U/L, q=1.22, P=0.824] between the HIRI IRF1 -/- group and the sham operation IRF1 -/- group. Compared with the HIRI group, the ratio of M1/M2-type Kupffer cells in the liver of the HIRI IRF1 -/- group decreased [(0.958±0.090) vs. (2.788±0.258), q=2.06, P<0.0001], and the mRNA expression of TNFα decreased [(4.363±0.393) vs. (12.900±5.504), q=5.59, P=0.018], and the differences between the two groups were statistically significant. In the co-culture and hypoxia-reoxygenation experiment using ANA1 cells overexpressing IRF1 and AML12 cells, the proportion of AML12 hepatocytes in late apoptosis was higher than that in the control group [(14.05±4.25) vs. (3.15±1.16), t=2.85, P=0.047], and the difference was statistically significant. In contrast, when the expression of IRF1 was knocked down, the proportion of apoptotic AML12 cells decreased [(9.26±3.04) vs. (13.36±4.64), t=2.15, P=0.098], but the difference was not statistically significant. Conclusion:The IRF1 protein can regulate the polarization of Kupffer cells into M1-type macrophages, promote the inflammatory injury of the liver tissue after ischemia-reperfusion, and increase the apoptosis of hepatocytes.

Objective To explore and analyze the related factors of kinesiophobia in elderly patients with coronary heart disease (CHD) during cardiac rehabilitation exercise period, and to evaluate the effect of cognitive behavior therapy on improving kinesiophobia and promoting rehabilitation. Methods A total of 352 elderly patients with CHD admitted to the hospital were included from October 2023 to October 2024. Tampa Scale for Kinesiophobia-11 (TSK-11) was adopted to evaluate the kinesiophobia status. Patients with kinesiophobia were randomly grouped. Routine intervention (routine group, n=82) and cognitive behavior intervention (study group, n=82) were implemented respectively. The intervention effects were observed in both groups. Results Among the 352 patients, 46.59 % (164/352 ) of elderly patients with coronary heart disease had different degrees of kinesiophobia. The proportions of female, divorced/widowed, revascularization and family relationship disharmony and scores of Patient Health Questionnaire (PHQ9) and Generalized Anxiety Disorder Scale (GAD7) in patients with kinesiophobia were higher than those in patients without kinesiophobia (P<0.05) while the scores of General Self-Efficacy Scale (GSES) and Social Support Rating Scale (SSRS) were lower compared with those in patients without kinesiophobia (P<0.05). Logistic regression analysis found that female, divorced/widowed, family relationship disharmony, revascularization and scores of PHQ9, TSK-11, GAD7, GSES and SSRS were related to kinesiophobia (P<0.05). After intervention, the scores of TSK-11, PHQ9 and GAD7 in the study group were lower while the scores of GSES and SSRS, 6 min walking test distance, and cardiopulmonary exercise test peak oxygen uptake and anaerobic threshold were higher compared to the routine group (P<0.05). Conclusion The kinesiophobia in elderly patients with CHD during cardiac rehabilitation is related to gender, revascularization and psychosocial factors. Clinically, cognitive behavior intervention should be provided according to the situation and guided to carry out rehabilitation exercise regularly so as to promote improvement of cardiopulmonary function.

The Healthy China 2030 Plan set the goal of reducing premature deaths from diabetes by 30% by 2030. However, there has been a lack of assessment of premature mortality for diabetes since the action plan was issued. This study used data from the Global Burden of Disease Study 2021, calculated the premature deaths for diabetes by sex, provinces, and subtypes from 1990 to 2021. We explored the temporal trend of premature mortality using the average annual percent change (AAPC) for different sexes, provinces, and subtypes from 1990 to 2021. Furthermore, we predicted premature mortality for diabetes through 2030 for China and its provinces according to the average annual change rate from 2010 to 2021. There was a first slow upward trend in premature mortality for diabetes from 0.5% in 1990 to 0.6% in 2004, and then a decline until 2021 with premature mortality of 0.4%. By 2030, only Fujian (30.3%) will achieve the desired level of reduction, with only seven provinces meeting the target for females and none for males. There is a large range in the degree of decline between inland and coastal regions, showing obvious geographic differences, and there should be a focus on balancing medical resources.
Humans ; China/epidemiology* ; Female ; Male ; Mortality, Premature/trends* ; Diabetes Mellitus/mortality* ; Goals ; Middle Aged ; Adult

Objective:To analyze the research status, hotspots, and trends of Banxia Houpu Decoction.Methods:Research literature about Banxia Houpu Decoction was retrieved from the CNKI, VIP, Wanfang Data, and SinoMed from January 1, 2000 to June 30, 2024. NoteExpress 3.6.0.9220 software was used to merge and eliminate duplicates, and CiteSpace 6.1.R6 software was used to draw and interpret the annual publication volume, authors, and keywords.Results:A total of 716 articles were included in this study, and the number of articles increased first and then decreased in recent years. The journal with the most published articles was Journal of Practical Traditional Chinese Medicine (23 articles); the author with the largest number of articles was Kong Lingdong from Nanjing University (7 articles); high-frequency keywords included "globus hystericus", "cough" and "chronic pharyngitis". A total of 11 labels were generated by clustering analysis, which could be summarized into two topics: clinical application (chronic pharyngitis, gastroesophageal reflux disease, aspiration pneumonia, depression, etc.) and literature type (clinical observation, experimental research). Conclusions:The research hotspots of Banxia Houpu Decoction are focused on clinical application and compatibility research. Using network pharmacology to predict potential drug targets and explore their mechanisms, conducting experimental research to explore pharmacological effects and clinical applications for treating diseases is a research trend in this field.

Objective To explore the application and effectiveness of the apprenticeship education model in Chinese herbal medicine teaching at Western medical college.Methods By comparing classic lecture-based teaching with a combined approach integrating apprenticeship education,the study assesses the impact on student learning outcomes.Participants included students from the 2018 cohort of the eight-year clinical medicine program and the 2022 cohort of the"4+4"pilot program at Peking Union Medical College,who received classic teaching methods and apprenticeship case-based teaching methods,respectively.Upon course completion,students completed a 14-item multiple-choice questionnaire covering essential theory of Chinese medicine,as well as specific categories such as qi-regulating,blood-activating herbs,among others.Results The overall accuracy rate in the apprenticeship case-based teaching group was significantly higher than that in the classic teaching group(P＜0.01).Conclusions The apprenticeship education model of Traditional Chinese Medicine has a positive effect on teaching of Chinese herbal medicine at West-ern medical college and warrants further promotion and application.

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 correlation and consistency of urine protein creatinine ratio (UPCR) with 24-hour urine protein quantification (24hUTP) in patients with lupus nephritis (LN).Methods:LN patients were enrolled from the STAR cohort of Peking University First Hospital, The time period is January 1, 2007 to July 31 2024. The correlation and consistency of UPCR with 24hUTP were analyzed in overall patients and various subgroups, such as,the levels of proteinuria and multiple parameters. The correlation was analyzed using Spearman′s method. The consistency was evaluated by using Kappa test, cross tabulation consistency probability, and UPCR/24hUTP ratio.Results:A total of 261 LN patients were included in this study. Overall, UPCR was positively correlated with 24hUTP ( r=0.892, P<0.001), and the consistency between then was good ( Kappa=0.603, P<0.001). Subgroup analysis based on multiple levels including different urine volume, eGFR grading, albumin, urea nitrogen, total cholesterol, low-density lipoprotein cholesterol, SLEDAI-2K, also revealed good correlation ( r=0.747~0.951; P<0.001) and consistency ( Kappa=0.393~0.771; P<0.001). Patients were divided into subgroups according to the 24hUTP (≤ 0.5 g, >0.5 g to ≤1.0 g, >1.0 g to ≤3.5 g, and>3.5 g). The best correlation and consistency were observed in ≤ 0.5g group ( r=0.728, Kappa=0.395, P<0.001), while the correlation was average or poor in other subgroups ( r=0.16~0.46). The consistency evaluated by UPCR/24hUTP ratio showed good consistency (ratios between 0.8~1.2) in 74 patients (28.4%), however, the unsatisfactory consistency in most patients, with 128 patients (68.4%) having ratios < 0.8, indicating a significantly lower UPCR than 24hUTP, and 59 patients (31.6%) having ratios > 1.2, indicating significantly higher UPCR than 24hUTP.The cross tabulation evaluation showed that the probability of consistency was distributed in a gradient, we found the worst consistency (48.9%) in >0.5 g to ≤1.0 g group, moderate consistency (64.0%) in >1.0 g to ≤3.5 g group, and the best consistency (73.3%) in >3.5 g group. Conclusion:In LN patients, the overall correlation and consistency between UPCR and 24hUTP are good, but the degree of correlation and consistency is closely related to the total amount of proteinuria. When 24hUTP ≤ 0.5 g, UPCR can be used to replace 24hUTP, but UPCR cannot be simply equated with 24hUTP at other urinary protein levels. Detecting 24hUTP to accurately reflect the total amount of proteinuria remain necessary.