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 compare the efficacy of controlling pediatric myopia progression between defocus incorpo-rated multiple segments(DIMS)spectacle lenses combined with low-concentration atropine(0.1 g·L-1)(DIMSA)and orthokeratology(OK)lenses.Methods A prospective cohort study was conducted on myopic children treated at the De-partment of Ophthalmology,Children's Hospital of Nanjing Medical University between January 2022 and February 2023.Participants were divided into a DIMSA group(88 cases)and an OK lens group(73 cases).Only right eye data were in-cluded for statistical analysis,and the children were followed up for 1 year.The axial length(AL)and spherical equivalent refraction(SER)were measured for the DIMSA group while only AL was measured for the OK lens group at baseline and each follow-up visit.The AL regression rate[(AL regression rate=the number of AL regression cases/total cases in the group)× 100％]and axial elongation(AE)were calculated to compare the myopia control efficacy between the two groups.Results The baseline age of the OK lens group was significantly larger than that of the DIMSA group(P=0.001).The AE value in the DIMSA group was significantly smaller than that in the OK lens group after following up for 6 months,following up for 1 year,and adjusting for baseline age(all P＜0.05).Multiple linear regression analysis revealed that baseline age was the only factor influencing the AE value[OK lens group:unstandardized β=-0.024,standardizedβ=-0.266(P=0.022);DIMSA group:unstandardized β=-0.052,standardized β=-0.487(P＜0.001)].There was a negative correlation between AE and baseline age(OK lens group:r=-0.30,P＜0.001;DIMSA group:r=-0.42,P＜0.001).In the DIMSA group,baseline age was positively correlated with SER progression(r=0.28,P=0.001 5).After following up for 1 year,the proportions of children with different AE velocities differed significantly between the two groups(x2=11.09,P=0.004).AE was controlled better in the DIMSA group than that in the OK lens group.AL regression rates in the DIMSA group were significantly higher than those in the OK lens group after 6 months and 1 year of follow-up(all P＜0.05).No severe ocular adverse events occurred in either group.Conclusion DIMSA demonstrates superior effi-cacy over OK lenses in controlling myopia progression in children with mild-to-moderate myopia.Ophthalmologists should tailor myopia control strategies based on individual needs and characteristics.

Objective To compare the efficacy of controlling pediatric myopia progression between defocus incorpo-rated multiple segments(DIMS)spectacle lenses combined with low-concentration atropine(0.1 g·L-1)(DIMSA)and orthokeratology(OK)lenses.Methods A prospective cohort study was conducted on myopic children treated at the De-partment of Ophthalmology,Children's Hospital of Nanjing Medical University between January 2022 and February 2023.Participants were divided into a DIMSA group(88 cases)and an OK lens group(73 cases).Only right eye data were in-cluded for statistical analysis,and the children were followed up for 1 year.The axial length(AL)and spherical equivalent refraction(SER)were measured for the DIMSA group while only AL was measured for the OK lens group at baseline and each follow-up visit.The AL regression rate[(AL regression rate=the number of AL regression cases/total cases in the group)× 100％]and axial elongation(AE)were calculated to compare the myopia control efficacy between the two groups.Results The baseline age of the OK lens group was significantly larger than that of the DIMSA group(P=0.001).The AE value in the DIMSA group was significantly smaller than that in the OK lens group after following up for 6 months,following up for 1 year,and adjusting for baseline age(all P＜0.05).Multiple linear regression analysis revealed that baseline age was the only factor influencing the AE value[OK lens group:unstandardized β=-0.024,standardizedβ=-0.266(P=0.022);DIMSA group:unstandardized β=-0.052,standardized β=-0.487(P＜0.001)].There was a negative correlation between AE and baseline age(OK lens group:r=-0.30,P＜0.001;DIMSA group:r=-0.42,P＜0.001).In the DIMSA group,baseline age was positively correlated with SER progression(r=0.28,P=0.001 5).After following up for 1 year,the proportions of children with different AE velocities differed significantly between the two groups(x2=11.09,P=0.004).AE was controlled better in the DIMSA group than that in the OK lens group.AL regression rates in the DIMSA group were significantly higher than those in the OK lens group after 6 months and 1 year of follow-up(all P＜0.05).No severe ocular adverse events occurred in either group.Conclusion DIMSA demonstrates superior effi-cacy over OK lenses in controlling myopia progression in children with mild-to-moderate myopia.Ophthalmologists should tailor myopia control strategies based on individual needs and characteristics.

Objective:To explore the effects of G protein-coupled receptor 55 (GPR55) antagonist CID16020046 on renal fibrosis in mice, and provide a new method and idea for the treatment of renal fibrosis.Methods:(1) GPR55 overexpression and GPR55 antagonist CID16020046 were used in renal fibroblasts (NRK-49F) of rats, respectively. Meanwhile,transforming growth factor-β1 (TGF-β1) was applied in the NRK-49F cells to observe the expression of fibrosis-related factors and inflammatory factors. (2) A mouse model of renal fibrosis with unilateral ureteral obstruction (UUO) was established in vivo. Eight-week-old male C57BL/6J mice (20-25 g) were randomly divided into three groups according to the random number table method: sham group ( n=6), model group (UUO group, n=7), model + CID16020046 drug (UUO+CID group, n=8). The drug CID16020046 (10 mg/kg) was intraperitoneally injected 1 day before modeling, on the day of modeling and every day after surgery in UUO+CID group, and the corresponding dose of 0.9% normal saline was injected intraperitoneally in sham and UUO groups.The mice were sacrificed for sampling 7 days after UUO surgery, and their renal function indicators, liver transaminase, and cardiac markers were examined. Western blotting and quantitative real-time PCR were used to detect the expression of renal fibrosis-related factors and inflammatory factors. Immunohistochemistry staining, Sirius red staining and Masson trichrome staining were used to detect the pathological changes of renal tissues. Results:(1) After NRK-49F cells were stimulated by TGF-β1, the mRNA and protein expression levels of GPR55 were significantly increased (both P<0.05). There was no statistically significant difference in the mRNA expression of fibrosis-related factors fibronectin and collagen Ⅰ, and inflammatory factors interleukin-1β and tumor necrosis factor-α between TGF-β1 group and TGF-β1 + GPR55 overexpression group (all P>0.05). Compared with the TGF-β1 group, the protein expression levels of fibrosis-related factors alpha-smooth muscle actin (α-SMA) and vimentin, and the mRNA expression levels of collagen Ⅰ and α-SMA were lower in the TGF-β1 + CID group (all P<0.05). (2) Compared with sham group, the mRNA and protein expression levels of GPR55 in UUO group were higher (both P<0.05). The serum creatinine in the UUO+CID group was lower compared to the UUO group ( P<0.05). There was no statistically significant difference in blood urea nitrogen, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and creatine kinase isoenzyme between UUO group and UUO+CID group (all P>0.05). Compared with the UUO group, the protein expression levels of renal fibrosis-related factors fibronectin, collagen Ⅰ and vimentin, and the mRNA expression levels of fibronectin, collagen Ⅰ, collagen Ⅲ and α-SMA were lower in the UUO+CID group (all P<0.05). The degree of renal tubular dilation and interstitial collagen fiber deposition in the UUO+CID group was significantly reduced compared to the UUO group (all P<0.05). Conclusions:CID16020046 can reduce serum creatinine in UUO mice, protect renal function, and simultaneously decrease the expression of fibrosis-related factors in renal fibroblasts and mouse kidney tissues, thereby alleviating renal fibrosis.

Objective:To figure out the association between the expression of m 6A RNA methylation regulators and the prognosis of children AML, and provide genetic markers for monitoring the progression and recurrence of AML. Methods:Twenty two m 6A RNA methylation regulators were firstly analyzed using the data from Therapeutically Applicable Research To Generate Effective Treatments(TARGET) database and The Genotype-Tissue Expression(GTEx) database, Wilcoxon rank test was performed to analyze the differentially expression of m 6A RNA methylation regulators between the AML and normal tissue, 296 AML children were divided into training cohort and validation cohort by simple random sampling method, Lasso regression was used to screen out the risk factors and the multivariate Cox regression was applied for establishing prognosis predicting model in training cohort. Kaplan-Meier survival curve, time-dependent ROC curve and multivariate Cox regression were used to estimate the efficiency of risk score calculated by predictive model in validation cohort. Results:Twenty one m 6A genes were up regulated in AML compared to Normal patients. Five m 6A RNA methylation regulators( ZC3H13, YTHDC2, HNRNPA2B1, METTL3, METTL5) were included in final predicting model. Risk score could independently predict the survival of AML patients in training cohort(HR:2.72, 95% CI: 1.54-4.81, P=0.000 6) and validation cohort(HR:2.01, 95% CI:1.14-3.50, P=0.016). Low-risk patients had better prognoses than high-risk patients both in training cohort( P=0.001 9) and validation cohort( P=0.023). Conclusion:This prognosis predicting model constructed by m 6A RNA methylation regulators could independently predict the survival prognosis in AML children, and should be helpful for clinical therapy.

Objective:To figure out the association between the expression of m 6A RNA methylation regulators and the prognosis of children AML, and provide genetic markers for monitoring the progression and recurrence of AML. Methods:Twenty two m 6A RNA methylation regulators were firstly analyzed using the data from Therapeutically Applicable Research To Generate Effective Treatments(TARGET) database and The Genotype-Tissue Expression(GTEx) database, Wilcoxon rank test was performed to analyze the differentially expression of m 6A RNA methylation regulators between the AML and normal tissue, 296 AML children were divided into training cohort and validation cohort by simple random sampling method, Lasso regression was used to screen out the risk factors and the multivariate Cox regression was applied for establishing prognosis predicting model in training cohort. Kaplan-Meier survival curve, time-dependent ROC curve and multivariate Cox regression were used to estimate the efficiency of risk score calculated by predictive model in validation cohort. Results:Twenty one m 6A genes were up regulated in AML compared to Normal patients. Five m 6A RNA methylation regulators( ZC3H13, YTHDC2, HNRNPA2B1, METTL3, METTL5) were included in final predicting model. Risk score could independently predict the survival of AML patients in training cohort(HR:2.72, 95% CI: 1.54-4.81, P=0.000 6) and validation cohort(HR:2.01, 95% CI:1.14-3.50, P=0.016). Low-risk patients had better prognoses than high-risk patients both in training cohort( P=0.001 9) and validation cohort( P=0.023). Conclusion:This prognosis predicting model constructed by m 6A RNA methylation regulators could independently predict the survival prognosis in AML children, and should be helpful for clinical therapy.

Objective To summarize the computed tomographic (CT) manifestations of pulmonary aspergillosis after organ transplantation and compare different signs between pulmonary aspergillosis and bacterial pneumonia.Methods CT images of pulmonary aspergillosis (n =62) and bacterial pneumonia (n =68) in post-transplantation patients were reviewed.The signs were categorized with consolidation,mass,large nodule (≥1crn),small nodule and bud-in-tree pattern.Some detailed useful differentiating signs such as halo sign,air bronchogram sign,reversed halo sign,hypodensity sign and cavitation were also analyzed.Results CT patterns of pulmonary aspergillosis included consolidation,mass,large nodule,small nodule and bud-in-tree pattern.The most common was large nodule (75.8％),followed by consolidation (48.4％)and mass (29.0％).And small nodule (16.1 ％) and bud-in-tree (12.9％) patterns were concurrent.For consolidation pattern,the proportion of bacterial pneumonia (69.1％) was the larger;For mass pattern,the proportion of pulmonary aspergillosis (29.0％) was the larger.For large nodule pattern,there was no difference.The detail sign of large nodule in two groups had no difference In detailed signs of consolidation pattern,air bronchogram sign was more often seen in bacterial pneumonia while cavitation was more frequently found in pulmonary aspergillosis.In detailed signs of mass pattern,pulmonary aspergillosis often has single lesion (66.7％),cavitation (83.3％)and air crescent sign (77.8％) is more common.The proportion of halo sign was 30.7％.Conclusions CT manifestations of pulmonary aspergillosis are diverse after organ transplantation.There is some difference and yet overlap with bacterial pneumonia.

Objective To investigate the expressions of spleen tyrosine kinase (Syk), c-Jun amino terminal kinase (JNK) and nucleotide binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in the heart tissue in SD rat model of diabetic cardiomyopathy, and to explore the relationship between Syk, JNK and NLRP3. Methods Clean male SD rats were randomly divided into the control (Ctrl) group and diabetic cardiomyopathy model (DCM) group. Rats of DCM group were treated with a single intraperitoneal injection of streptozotocin (STZ), while rats of Ctrl group were injected with the same dose of citrate buffer. The random blood glucose level and body weight were monitored every week until 20 weeks after STZ or citrate buffer injection, then all the rats were killed and their hearts were obtained. Rat H 9c2 cardiomyocytes were randomly divided into normal glucose treatment (NG) group, high glucose treatment (HG) group, Syk inhibitor control (BAY) group and Syk inhibitor high glucose (HG+BAY) group. The Syk and JNK phosphorylations and NLRP3 protein expression were detected by Western blot assay in the heart tissue of SD rats and H9c2 cardiomyocytes. The NLRP3, cysteine-containing aspartate specific protease 1(caspase-1) and interleukin (IL)-1β expressions at mRNA level were detected by reverse transcription-polymerase chain reaction (RT-PCR). Results The random blood glucose level was significantly increased (P<0.05) and the body weight was significantly decreased (P<0.05) in DCM group compared with those of Ctrl group. The expressions of cardiac p-Syk, p-JNK and NLRP3 at protein level were significantly increased in DCM group compared with those of Ctrl group (P<0.05). Furthermore, the mRNA levels of NLRP3, caspase-1 and IL-1β were significantly up-regulated (P < 0.05). BAY treatment significantly inhibited the high glucose-induced NLRP3, caspase-1 and IL-1β mRNA expressions and p-JNK, NLRP3 protein expressions in H9c2 cardiomyocytes (P < 0.05). Conclusion JNK phosphorylation and NLRP3 inflammasome activation induced by Syk play an important role in the pathogenesis of diabetic cardiomyopathy.

Objective To summarize the diagnostic efficacy of ADC value for differentiation of benign and malignant lymph nodes on diffusion MRI with Meta-analysis. Methods Published papers on differentiation of benign and malignant lymph nodes with ADC value were searched and reviewed.Quality evaluation was performed for the eligible papers before data extraction.Test for heterogeneity was performed first,then appropriate model was selected to calculate the weighted mean difference,sensitivity,specificity,positive likelihood ratio,negative likelihood ratio,diagnostic odds ratio,pretest and posttest probability.The potential of ADC value for differentiation of benign and malignant lymph nodes was assessed qualitatively and quantitatively.Results Fifteen papers including 735 cases and 1963 lymph nodes were selected.According to Meta-regression analysis,subgroup analysis and robust analysis,two studies with benign lymph nodes in patients with benign lesion and one study using chemical shift saturation technique were excluded because of their impact on the robustness of the pooled results. The weighted mean difference (WMD) between malignant and benign lymph nodes was -0.355 × 10-3mm2/s[95％ confidence interval (CI):-0.423 ×10-3- -0.288 × 10-3 mm2/s].Although the cutoff of ADC value for differentiation in each study was different,the diagnostic efficacy was stable,the pooled sensitivity,specificity,positive likelihood ratio,negative likelihood ratio,diagnostic odds ratio and area under summarized receiver operator's curve were 0.87 (95％ CI:0.79-0.92),0.87 (95％ CI:0.82-0.90),6.5 (95％ CI:4.7-9.2),0.15 (95％ CI:0.09-0.25 ),43 ( 95％ CI:21-87 ),0.93 ( 95 ％ CI:0.90-0.95 ).The posttest malignancy probability of benign lymph node indicated by ADC was 6％,while that of malignant lymph node was 72％.Conclusion The ADC value can be used to differentiate benign and malignant lymph nodes with good sensitivity and specificity noninvasively.