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 explore the correlation between childhood obstructive sleep apnea hypopnea syndrome and otitis media with effusion,to provide evidence-based basis for clinical diagnosis and treatment.METHODS A total of 985 children were diagnosed in the Department of Otolaryngology,Heping Hospital Affiliated to Changzhi Medical College from September 2019 to March 2025.Combined or no OME patients for multivariate logistics regression analysis and construct related risk prediction model,obtain data of OSAHS(exposure factors)and OME(outcome)from the public database of genome-wide association studies(GWAS),two-sample Mendelian randomization(MR)analysis.RESULTS The risk prediction model for OME in children with OSAHS,Hosmer-Lemeshow goodness of fit test(χ2=12.541,P=0.157),C index of 0.749,Calibration calibration curve fitting,and correlation between children OSHAS and OME.Furthermore,the MR analysis using inverse variance weighting observed a positive causal relationship between OSAHS and OME in children(OR=1.05,95%CI=0.999-1.051,P=0.005).MR Egger test results(Q=42.358,P=0.716)and IVW(Q=43.271,P=0.759)showed no heterogeneity in the analysis,MR Egger intercept(intercept=-0.004,P=0.921)showed no horizontal pleiotropy,and the MR analysis results were stable.CONCLUSION There is a correlation between pediatric OSAHS and OME,and it increases the risk of OME.

Objective:To establish and validate a nomogram for predicting the probability of central lymph node metastasis (CLNM) by evaluating risk factors in clinical N 0(cN 0) stage patients with iisolated papillary thyroid carcinoma of isthmus (PTCI). Methods:Retrospectively analyzed the clinicopathological data of 112 PTCI patients who underwent surgical treatment at Lianyungang Municipal Oriental Hospital between January 2019 and December 2024. There were 35 males and 77 females, with an average age of (47.2±12.6) years, ranging from 28 to 71 years. Univariate and multivariate logistic regression analyses were performed to identify independent risk factors for CLNM. Nomogram method was used to build the transfer risk prediction model and verify it.Results:The study enrolled 112 patients, with 42 cases (37.5%) exhibiting CLNM. ROC curve analysis demonstrated that tumor diameter predicted CLNM with an AUC of 0.781 (95% CI: 0.694-0.868), identifying an optimal cutoff value of 0.9 cm (sensitivity: 0.762, specificity: 0.700). Multivariate logistic regression revealed that CLNM was independently associated with age, tumor diameter, capsular invasion, and Hashimoto′s thyroiditis (HT), all P<0.05. The logistic regression analysis results were used to construct the nomogram model of CLNM risk prediction. Hosmer lemeshow test resulted χ2=6.876, P=0.442. The calibration curves of the model group and the validation group were basically close, with an average absolute error of 0.065, P=0.099. ROC curve analysis showed that the best cut-off value of nomogram was 160 points, C-index was 0.849 (95% CI: 0.771-0.927), sensitivity was 0.786, specificity was 0.729. Decision curve analysis (DCA) based on net benefit and threshold probability shows that the nomogram can obtain net benefit in a large threshold probability range (0.13-1.00). Temporary recurrent laryngeal nerve injury occurred in 5 patients (4.5%). Temporary parathyroid injury occurred in 4 cases (3.6%), all of which were normal within one month. Conclusions:For patients with cN 0 stage isolated PTCI, CLNM is related to age ≤45 years, tumor diameter>0.9 cm, envelope invasion and HT. HT is a protective factor for CLNM. The ROC curve, calibration curve and DCA curve verified internally showed that the nomogram obtained had good discrimination ability, calibration ability and clinical applicability. When the total score ≥ 160 points indicates that the risk of CLNM is increased.

Objective:To evaluate the clinical efficacy of Sanren Decoction combined with triple therapy in the treatment of Hp-related chronic atrophic gastritis (CAG).Methods:A randomized controlled trial was conducted. A total of 118 patients with Hp-related CAG admitted to Department of Spleen and Stomach Diseases of Xi'an Affiliated Hospital of Shaanxi University of Chinese Medicine from February 2021 to February 2023 were selected as observation subjects, and were divided into two groups according to random number table method, with 59 cases in each group. The control group received triple therapy (amoxicillin+clarithromycin+omeprazole), whereas the observation group was combined with modified Sanren Decoction on the basis of the control group. Both groups were continuously treated for 10 days. TCM symptoms were scored before and after treatment. Updated Sydney System (UAS) was applied to evaluate the gastric mucosal lesions. ELISA was applied to detect serum levels of cluster of differentiation 47 (CD47), VEGF and pepsinogenⅠ/pepsinogen Ⅱ (PGⅠ/PGⅡ). The Hp infection status was evaluated by Urea Breath Test (UBT). In addition, the adverse reactions were observed and recorded during treatment.Results:After treatment, the scores of gastric fullness, stomachache, acid reflux and bitter taste in observation group were lower than those in the control group ( t=4.52, 3.46, 4.34, 4.83, P<0.01), and the scores of gastric mucosal inflammation, atrophy, intestinal metaplasia and dysplasia were also lower compared to control group ( t=3.68, 2.88, 3.73, 4.45, P<0.01 or P<0.05). After treatment, the levels of serum CD47 [(4.46±1.29) μg/L vs. (5.01±1.37) μg/L, t=2.25] and VEGF [(70.53±10.47) ng/L vs. (79.34±10.56) ng/L, t=4.55] in observation group were lower ( P<0.05 or P<0.01), while the level of PGⅠ/PGⅡ [(12.45±1.78) vs. (10.82±1.62), t=5.20] was higher compared with that of the control group ( P<0.01). After treatment, the negative conversion rate of Hp was 96.61% (57/59) in observation group and that in control group was 81.36% (48/59) ( χ2=7.00, P=0.008). During treatment, the incidence of adverse reactions was 23.73% (14/59) in observation group and 15.25% (9/59) in control group, revealing no statistical significance between the two groups ( χ2=1.35, P=0.245). Conclusion:Modified Sanren Decoction combined with triple therapy can improve gastric mucosal lesions, reduce serum CD47 and VEGF levels, and increase PGⅠ/PGⅡ ratio and Hp negative conversion rate in patients with Hp-related CAG, with good safety.

Objective To investigate the immediate therapeutic efficacy of Kuanxiong Aerosol on improving the angina pectoris in the patients complicated with intermediate coronary stenosis(ICS),and to observe its effect on resting full-cycle ratio(RFR),corrected TIMI(thrombolysis in myocardial infarction)frame count(CTFC)in angiography,and coronary serum inflammatory factors.Methods Sixty angina pectoris patients with ICS admitted to the Cardiovascular Department of Dade Road Hospital,Guangdong Provincial Hospital of Traditional Chinese Medicine from March 2023 to March 2024 were randomly divided into the trial group and the control group,with 30 patients in each group.The trial group was given four consecutive sprays of Kuanxiong Aerosol by sublingual spray,and the control group had no intervention but just was given the monitoring for 10 minutes.Before and after the intervention,the changes of coronary RFR,CTFC,Visual Analogue Scale(VAS)score of chest pain,and the serum levels of C-reactive protein(CRP),interleukin 6(IL-6)and lipoprotein-associated phospholipase A2(Lp-PLA2)in the two groups were observed.Moreover,the incidence of adverse reactions during the intervention in the two groups of patients was compared.Results(1)After the intervention,the coronary RFR value of the trial group was increased significantly compared with that before intervention(P<0.01),while the coronary RFR value of the control group was not increased significantly compared with that before intervention(P>0.05);the comparison between the two groups showed that the effect on increasing the coronary RFR value in the trial group was superior to that in the control group(P<0.05).(2)After intervention,the CTFC value of the trial group was significantly decreased compared with that before intervention(P<0.01),while the CTFC value of the control group was not significantly decreased compared with that before intervention(P>0.05);the intergroup comparison showed that the trial group tended to have a better effect on the decrease of CTFC value than the control group,but the difference being not statistically significant(P>0.05).(3)After the intervention,the chest pain VAS score of the trial group was significantly reduced compared with that before intervention(P<0.01),while the pre-and post-treatment changes of the score in the control group was not significant(P>0.05);the intergroup comparison showed that the decrease of the chest pain VAS score in the trial group was superior to that in the control group(P<0.01).In particular for immediate therapeutic efficacy,Kuanxiong Aerosol achieved the effective rate of 96.67%(29/30)for relieving chest pain 10 minutes after sublingual spraying,which was significantly superior to that of the control group[10.00%(3/30)],and the comparison between the two groups showed that the difference was statistically significant(P<0.001).(4)After the intervention,the Lp-LPA2 value of the trial group was decreased compared with that before intervention(P<0.05),while the CRP and IL-6 values of the trial group as well as the CRP,IL-6,and Lp-LPA2 values of the control group were all not significantly decreased compared with those before intervention(P>0.05).The intergroup comparison showed that the trial group's effect on the decrease of Lp-LPA2 value was significantly superior to that of the control group(P<0.05).(5)Before and after the intervention,no obvious changes of the general vital signs in the two groups were shown,no drug-related adverse occurred,either.Conclusion Kuanxiong Aerosol can immediately improve the coronary physiological function indicators of angina pectoris patients with ICS,increase the coronary flow rate,and inhibit inflammatory response of the coronary artery to some degree,thus to alleviate the symptoms of angina pectoris in patients with ICS.

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

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.

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.