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 epidemiology of elderly respiratory tract infection (RTI) cases in a hospital in Xuzhou region from 2020 to 2022. Methods The cases of RTI patients in a hospital were screened from May 2020 to December 2022, and 548 cases that met the criteria were included in the study. Patient case data were analyzed for symptoms, pathogen distribution, and differences in patient distribution under different screening conditions (age, disease, and season). Results More than 90.00% of the included RTI patients presented with symptoms of cough, sputum, wet rales and pleural effusion was less common. The top three comorbidities were cardiovascular disease (153 patients, 27.92%), cerebrovascular disease (133 patients, 24.27%), and gastrointestinal disease (105 patients, 19.16%).All 548 elderly patients tested positive for respiratory pathogens (100.00%). There were 540 cases of single pathogen infection (98.54%) and 8 cases of mixed infection (1.46%). The top five single pathogen infections were Pseudomonas aeruginosa (92 cases, 16.76%), Escherichia coli (78 cases, 14.21%), drug-resistant Staphylococcus aureus (69 cases, 12.57%), Klebsiella pneumoniae (65 cases, 11.84%), and Mycoplasma pneumoniae (46 cases, 8.38%). The highest detection rate of respiratory pathogens was found in patients >90 years old, whose main pathogens were Pseudomonas aeruginosa, Klebsiella pneumoniae and drug-resistant Staphylococcus aureus. The next highest rates of pathogen detection were found in patients aged 86-91 and 81-85 years, unlike patients >90 years, who had a higher rate of Escherichia coli detection. Unlike other age groups, patients <75 years old had a higher percentage of influenza B virus detection. The highest incidence of pneumonia was found in 45.62% (250 cases). Escherichia coli had the highest detection rate in acute bronchitis/episodes and pneumonia, respiratory syncytial virus had the highest detection rate in wheezing bronchitis, Klebsiella pneumoniae had the highest detection rate in bronchopneumonia, and Pseudomonas aeruginosa had the highest detection rate in fever. The highest detection rate of pathogens was found in fall (36.50%), followed by spring (27.01%). The distribution of pathogen infections in all seasons was matched with the results of pathogenicity testing. Streptococcus oxysporus had the highest number of infections in the fall (χ²=20.33, P<0.001). Conclusion Elderly respiratory tract infections in this region are most common in patients over 90 years old, with the highest incidence of pneumonia and high incidence in fall, and the pathogens are mainly Pseudomonas aeruginosa, Escherichia coli and drug-resistant Staphylococcus aureus. Attention to distinguish the above characteristics can provide some support for early diagnosis and treatment of respiratory infections in the elderly in this region.

AIM: To investigate the role of pyroptosis in the development of diabetic retinopathy(DR)and to explore the regulatory mechanism by which circular RNA(circRNA)and its targeted microRNA(miRNA)mediate pyroptosis in DR, providing new therapeutic targets and strategies for the prevention and treatment of DR.METHODS: A streptozotocin(STZ)-induced model of type 1 diabetes in SD rats was established. The expression of circRNA_0076631, miR-214, and pyroptosis-related factors were measured in retinal tissues. CCK-8 and tube formation assays were used to detect the effect of different concentration of glucose on cell proliferation and angiogenic abilities of human retinal microvascular endothelial cells(HRMECs). The expression levels of circRNA_0076631, miR-214, and pyroptosis-related markers were evaluated through qRT-PCR and Western blot analysis, with additional experiments conducted following circRNA_0076631 knockdown to assess its effect on pyroptosis markers. Previous bioinformatics analysis and luciferase reporter assays identified a shared binding site among circRNA_0076631, miR-214, and caspase-1. To clarify the interaction between these molecules, co-transfection experiments using circRNA_0076631 inhibitors(ASO-circRNA_0076631), miR-214 overexpression transfection reagent, and miR-214 inhibitors(AMO-miR-214)were conducted to elucidate the regulatory pathway involved in DR.RESULTS: Both the diabetic rat model and D-glucose-treated HRMECs showed significantly elevated expression of circRNA_0076631 and pyroptosis-related factors(NLRP3, caspase-1, and IL-1β), while miR-214 expression was reduced(all P<0.05). The mRNA expression of pyroptosis-related factors caspase-1 was reduced after the overexpression of miR-214, and it was upregulated after the inhibition of miR-214(all P<0.05). Knockdown of circRNA_0076631 reduced the mRNA expression of pyroptosis markers caspase-1(P<0.05). Co-transfection experiments revealed that the inhibition circRNA_0076631 suppressed pyroptosis(all P<0.05), but this suppression was reversed upon co-transfection with miR-214 inhibitors, leading to increased mRNA expression of the pyroptosis marker caspase-1(all P<0.05).CONCLUSION: The circRNA_0076631 and pyroptosis play critical roles in the pathogenesis of DR, and circRNA_0076631 may regulate pyroptosis by modulating miR-214, which in turn influences the expression of caspase-1 in the pyroptosis signaling pathway, thereby contributing to DR progression. The circRNA_0076631 may serve as a novel therapeutic target, providing new insights for the prevention and treatment of DR.

AIM: To investigate the role of pyroptosis in the development of diabetic retinopathy(DR)and to explore the regulatory mechanism by which circular RNA(circRNA)and its targeted microRNA(miRNA)mediate pyroptosis in DR, providing new therapeutic targets and strategies for the prevention and treatment of DR.METHODS: A streptozotocin(STZ)-induced model of type 1 diabetes in SD rats was established. The expression of circRNA_0076631, miR-214, and pyroptosis-related factors were measured in retinal tissues. CCK-8 and tube formation assays were used to detect the effect of different concentration of glucose on cell proliferation and angiogenic abilities of human retinal microvascular endothelial cells(HRMECs). The expression levels of circRNA_0076631, miR-214, and pyroptosis-related markers were evaluated through qRT-PCR and Western blot analysis, with additional experiments conducted following circRNA_0076631 knockdown to assess its effect on pyroptosis markers. Previous bioinformatics analysis and luciferase reporter assays identified a shared binding site among circRNA_0076631, miR-214, and caspase-1. To clarify the interaction between these molecules, co-transfection experiments using circRNA_0076631 inhibitors(ASO-circRNA_0076631), miR-214 overexpression transfection reagent, and miR-214 inhibitors(AMO-miR-214)were conducted to elucidate the regulatory pathway involved in DR.RESULTS: Both the diabetic rat model and D-glucose-treated HRMECs showed significantly elevated expression of circRNA_0076631 and pyroptosis-related factors(NLRP3, caspase-1, and IL-1β), while miR-214 expression was reduced(all P<0.05). The mRNA expression of pyroptosis-related factors caspase-1 was reduced after the overexpression of miR-214, and it was upregulated after the inhibition of miR-214(all P<0.05). Knockdown of circRNA_0076631 reduced the mRNA expression of pyroptosis markers caspase-1(P<0.05). Co-transfection experiments revealed that the inhibition circRNA_0076631 suppressed pyroptosis(all P<0.05), but this suppression was reversed upon co-transfection with miR-214 inhibitors, leading to increased mRNA expression of the pyroptosis marker caspase-1(all P<0.05).CONCLUSION: The circRNA_0076631 and pyroptosis play critical roles in the pathogenesis of DR, and circRNA_0076631 may regulate pyroptosis by modulating miR-214, which in turn influences the expression of caspase-1 in the pyroptosis signaling pathway, thereby contributing to DR progression. The circRNA_0076631 may serve as a novel therapeutic target, providing new insights for the prevention and treatment of DR.

Objective: To construct a prediction model for sleep disorders in shift workers of a chemical fiber enterprise, so as to provide the basis for early identification and prevention of sleep disorders in shift workers. Methods: Shift workers were sampled from a chemical fiber enterprise in Tongxiang City, Zhejiang Province using a cluster sampling method from August 2022 to July 2024. Demographic information, length of service and average weekly working hours were collected through questionnaire surveys. Depressive symptoms, anxiety symptoms and sleep disorders were evaluated using the Pittsburgh Sleep Quality Index, Patient Health Questionnaire and Generalized Anxiety Disorder Questionnaire, respectively. The shift workers were randomly divided into a training set and a validation set at a ratio of 7∶3. Predictive factors were selected using a multivariable logistic regression model based on the training set, and a nomograph model for prediction of sleep disorders in shift workers was established. The predictive values of the model were evaluated using the receiver operating characteristic (ROC) curve and calibration curve based on the training set and validation set. Results: Totally 673 shift workers were included, with a median age of 32 (interquartile range, 12) years. There were 493 males, accounting for 73.25%. There were 471 (69.99%) workers in the training set and 202 (30.01%) workers in the validation set. There were 274 workers with sleep disorders, accounting for 40.71%. The equation for the prediction model was ln[p/(1-p)]=-8.391+1.906×average weekly working hours+1.822×depressive symptoms+1.667×anxiety symptoms. The area under the ROC curve was 0.769 (95%CI: 0.661-0.835) for the training set and 0.655 (95%CI: 0.593-0.737) for the validation set, and Hosmer-Lemeshow test showed a good fitting effect (both P>0.05). Conclusion The nomograph model constructed by average weekly working hours, depressive symptoms and anxiety symptoms can be used to predict the risk of sleep disorders in shift workers of a chemical fiber enterprise.

BACKGROUND:Studies have reported that alendronate intake significantly increases bone mineral density in patients with osteoporosis. OBJECTIVE:To analyze and compare the changes in metabolites before and after alendronate intervention in ovariectomized rats by chromatography-mass spectrometry,and to further explore the specific mechanism and target of alendronate in the treatment of osteoporosis. METHODS:A total of 36 female Sprague-Dawley rats were randomly divided into model group,alendronate sodium group and sham operation group.The osteoporosis model was established by ovariectomy in the first two groups.Four weeks after modeling,the rats in the alendronate group were intragastrically given alendronate sodium,while those in the sham operation group and model group were given equal volume of normal saline.After 12 weeks of continuous gavage,the metabolites of the lumbar spine were analyzed by chromatography-mass spectrometry,and the common differential metabolites were obtained,which were analyzed by bioinformatics such as Kyoto Gene and Genome Encyclopedia pathway. RESULTS AND CONCLUSION:Totally 17 different metabolites were obtained in the three groups.The enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes showed that alendronate sodium could regulate unsaturated fatty acid biosynthesis,linoleic acid metabolism and other pathways to protect ovariectomized rats.These results indicate that alendronate sodium may exert its anti-osteoporosis effect by interfering with unsaturated fatty acid bioanabolism and linoleic acid metabolism,so as to achieve the purpose of preventing osteoporosis

Chemical investigation of the stems of Fritillaria unibracteata P.K. Hsiao & K.C. Hsia resulted in the isolation of nine compounds, by means of silica gel column chromatography, and preparative HPLC. Based on spectroscopic and chemical evidence, these compounds were identified as: 27-hydroxychlorogenone (1), sieboldogenin (2), (3β, 25S)-spirost-5-ene-3,17,27-triol (3), laxogenin (4), tigogenone (5), cerevisterol (6), ergosterol peroxide (7), stigmaterol (8), and β-sitosterol (9). Compound 1 was a new compound, and compounds 2-9 were isolated from the stems of Fritillaria unibracteata for the first time. The inhibitory effects of compounds 1−9 on A549 cells were determined using the MTT method. The results show that compound 6 exhibits moderate inhibitory activity with an IC50 value of (14.16 ± 1.11) μmol/L.

The transition of cancer cells from epithelial state to mesenchymal state awarded hepatocellular carcinoma (HCC) stem cell properties and induced tumorigenicity, drug resistance, and high recurrence rate. Reversing the mesenchymal state to epithelial state by inducing mesenchymal-epithelial remodeling could inhibit the progression of HCC. Using high-throughput screening, chrysin was selected from natural products to reverse epithelial-mesenchymal transition (EMT) by selectively increasing CDH1 expression. The target identification suggested chrysin exerted its anti-HCC effect through covalently and specifically binding threonine 205 (Thr205) of alpha-enolase (ENO1). For the first time, we revealed that ENO1 bound β-catenin mRNA, and recruited YTHDF2 to identify the m6A modified β-catenin in the 3'-UTR region to degrade β-catenin mRNA. Eventually, the CDH1 gene expression was improved through the regulation of β-catenin mRNA. ENO1/β-catenin mRNA interaction might be a promising target for cellular plasticity reprogramming. Moreover, chrysin could mediate mesenchymal‒epithelial remodeling through increasing degradation of β-catenin mRNA by promoting the binding of ENO1 and β-catenin mRNA. To the best of our knowledge, chrysin is the first reported small molecule inducing β-catenin mRNA degradation through binding to ENO1. The water-soluble derivative of chrysin may be a natural product-derived lead compound for circumventing metastasis, recurrence, and drug resistance of HCC by mediating mesenchymal‒epithelial remodeling.

OBJECTIVES: To analyze the molecular mechanism of Xixian Pills for treatment of rheumatoid arthritis (RA). METHODS: Forty-eight rats were randomized into 6 groups (n=8), including a normal control group, a collagen-induced arthritis (CIA) model group, 3 Xixian Pills treatment (200, 400 and 800 mg/kg) groups, and a Tripterygium glycosides tablet (TGT) treatment group. In the latter 4 groups, the rats were treated with daily gavage of Xixian Pills or TGT 2 weeks after CIA modeling for 3 consecutive weeks. The differentially expressed proteins in high-dose Xixian Pills group and the model group compared with the normal control group were screened based on the tandem mass spectrometry tag (TMT) technology, and the core targets and signaling pathways were analyzed. The immune cell infiltration and gene expression data were analyzed using ggplot2 and tidyverse packages, and the correlation coefficients between the core targets and the immune cells were calculated. RESULTS: The CIA rats showed significantly increased serum levels of TNF-α and IL-6 and lowered serum IL-10 level. Treatments with high- and medium-dose Xixian Pills and TGT all significantly reduced serum TNF‑α and IL-6 and increased IL-10 levels in CIA rats. Proteomic analysis identified 160 differential proteins between the model group and high-dose Xixian Pills group, and the core targets included CCL5, STAT1, GZMB and IL7R. The areas under the ROC curve of CCL5 and STAT1 were both greater than 0.9. Immunohistochemical and immunofluorescence staining revealed increased levels of CCL5 and STAT1 in the ankle joints of CIA rats, which were significantly decreased after treatment with Xixian Pills. CONCLUSIONS Treatment with Xixian Pills offers protection of the joints in CIA rats possibly by inhibiting joint inflammation via regulating protein expressions of CCL5 and STAT1.
Animals ; Drugs, Chinese Herbal/pharmacology* ; Rats ; Arthritis, Rheumatoid/metabolism* ; Proteomics ; Tripterygium/chemistry* ; Arthritis, Experimental/metabolism* ; Tumor Necrosis Factor-alpha/blood* ; Interleukin-10/blood* ; Interleukin-6/blood* ; Male ; Rats, Sprague-Dawley ; Signal Transduction

Narrative is the cornerstone of interpersonal relationships and life safety. However， its important value in daily life， school education， health management， personal happiness， career development， and other aspects has been ignored. The narrative ecology of families， schools， hospitals， workplaces， and elderly care institutions is worrying， the narrative connection between parent-child and intergenerational is broken， the narrative nature of adolescents is ignored， the narrative demands of patients are neglected， narrative relationships in the workplace are indifferent， and the narrative capital of the elderly is idle. These issues have resulted in serious social problems， such as depression and suicide among adolescents， conflicts between doctors and patients， workplace and life burnout among middle-aged people， and the inability of the elderly to achieve healthy aging， which have become a “stumbling block” to the realization of holistic health. Advocating the construction of narrative ecology and interpersonal narrative connections is an important measure of achieving holistic health. Taking the “narrative concept” as the overall framework， and based on the research， education， and practice carried out by the Alliance of Narrative Medicine in Higher Education Institutions， this paper proposed that actively build China’s narrative system of life and health， to enable narrative play an active and dynamic role in the construction of narrative ecology in different spaces， such as the families， the schools， the hospitals， the workplaces， and the elderly care institutions， as well as practically improve the quality of life of the people.