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 epidemiological characteristics of respiratory virus infections in Kunming area during 2023-2024,and explore the detection rates of different virus types and their distribution patterns across different time periods and age groups.Methods A total of 10354 nasopharyngeal swab or sputum samples were retrospectively collected from patients with acute respiratory infections who visited the Third People's Hospital of Kunming City between March 2023 and June 2024.Multiple pathogens were detected using real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)technology.A retrospective analysis was then conducted on the clinical laboratory detection results,statistically analyzing the overall detection rates of various respiratory viruses,multiple infection phenomena,gender differences,age distribution,seasonal variations,infection site differences,and monthly infection situations.Results Among the 10354 patient respiratory samples tested,3368 pathogen infections were detected,with a detection rate of 32.53%(3368/10354).204 patients presented with mixed infections of≥2 pathogens,with a mixed detection rate of 6.06%(204/3368).The single detection rate for females was significantly higher than males(P<0.001),and the multiple infection detection rate for males is significantly higher than females(P<0.05),indicating that males may have a higher risk of concurrent infections.Among different age groups,the virus detection rate was highest in the 5-18 years age group at 55.87%.Significant differences were observed in the detection rates of FluA,FluB,and SIV-H3 across different disease types(P<0.05).In March 2023,the detection rate was highest at 54.27%(5619/10354),with Influenza A virus(FluA)and seasonal influenza H3 subtype(SIV-H3)being the most detected pathogens.Conclusion In acute respiratory infection(ARI)cases in the Kunming area,FluA,FluB,and SIV-H3 were the primary viral pathogens,with the region's viral epidemic characteristics closely related to patient age stages,seasonal changes,and infection site factors.

Objective To evaluate the efficacy and safety of polyethylene glycol electrolyte powder(PEG)combined with linaclotide(Lin)for bowel preparation in elderly constipated patients before colonoscopy.Methods In this prospective,randomized controlled trial,90 elderly patients with constipation undergoing colonoscopy were recruited at our hospital from June 2022 to December 2023.Participants were randomly assigned to three groups(n=30 each):PEG-3L alone,PEG-3L+Lin,and PEG-2L+Lin.Primary outcome was Boston Bowel Preparation Scale(BBPS)score and secondary outcomes included adverse event rates,colonoscopy completion rate,withdrawal time,and polyp detection rate.Statistical analysis was performed using independent t-tests and chi-square tests.Results The PEG-3L+Lin group achieved significantly higher BBPS scores than both PEG-3L alone and PEG-2L+Lin groups did(both P<0.001).The PEG-2L+Lin group also outperformed the PEG-3L alone group in cleansing efficacy(90.0%vs.76.7%,P=0.008).The PEG-2L+Lin group demonstrated the best tolerability and lowest adverse event rate,the PEG-3L group had the longest withdrawal time(P<0.05),but the three groups showed no significant difference in polyp detection rates.Conclusion PEG combined with linaclotide significantly improves bowel cleansing in elderly constipated patients.PEG-2L+Lin regimen provides optimal balance between efficacy,safety,and tolerability,making it a preferable choice for this population.

Objective To investigate the application of operation plan based on injury stress sequence in the treatment of degree Ⅲ and Ⅳ ankle fractures with Lange-Hansen classification of pronation and external rotation.Methods From May 2019 to May 2022,80 eligible patients of degree Ⅲand Ⅳ ankle fractures with Lange-Hansen classification were selected prospectively and randomely divided into 2 groups:40 patients in the study group were treated with surgical reduction according to the sequence of injury stress,and the other 40 patients in the control group were treated with traditional reduction sequence.The operative time,intraoperative blood loss,fracture healing time,full weight-bearing time,Ankle dorsoextension Angle,plantar flexion Angle,and American Orthopaedic Foot and Ankle Association were compared between the two groups Society,AOFAS)Ankle and posterior foot function scores and complications.Results The surgical time[(44.80±5.12)min vs.(47.61±6.42)min]and surgical bleeding volume[(43.81±9.02)ml vs.(51.50±12.01)ml]between the study group and the control group were statistically significant(all P＜0.05).Before surgery,there had no significant differences in ankle joint dorsiflexion angle,plantar flexion angle,AOFAS ankle and hind foot function score in 2 groups(all P＞0.05).The ankle dorsiflexion angle at 3 months after surgery[(16.84±1.77)° vs.(15.50±1.65)°],plantar flexion angle at 3 months after surgery[(45.64±4.10)° vs.(42.58±3.86)°]and at 6 months after surgery[(52.38±3.03)°vs.(50.64±3.74)°],the maximum walking distance score[(3.50±0.64)vs.(3.16±0.60)],ground walking score[(2.06±0.56)vs.(1.72±0.48)]and ankle joint anteroposterior activity score at 3 months after surgery[(4.96±1.38)vs.(4.16±1.07)]of the study group were all higher than those in the control group(all P＜0.05).There was no statistically significant difference in terms of fracture healing time,complete weight-bearing time,postoperative pain score,ankle joint function autonomous activity support score,abnormal gait score,maximum walking distance score at 6 months after surgery,ground walking score,ankle joint anterior and posterior activity score and surgical complications between 2 group(all P＞0.05).Conclusion According to the sequence of injury stress,surgical reduction for the treatment of grade Ⅲ and Ⅳ ankle fractures of Lange-Hansen classification of middle pronation and external rotation can shorten the operation time and promote the early recovery of joint function.

Objectives:To assess the impact of intravascular ultrasound(IVUS)guidance for drug-eluting stent(DES)implantation on the long-term outcomes in coronary artery disease(CAD)patients with chronic kidney disease(CKD).Methods:A retrospective study was conducted on 1 800 CAD and CKD patients who underwent coronary DES implantation at Nanjing First Hospital from January 2018 to December 2022.Patients were divided into IVUS-guided group(IVUS group,n=333)and angiography-guided group(angiography group,n=1 467).Propensity score matching(PSM)was performed at a 1:2 ratio to adjust for differences in baseline clinical and angiographic characteristics between the two groups.Major adverse cardiovascular events(MACE),including cardiac death,target vessel myocardial infarction,and clinically driven target vessel revascularization,were evaluated over a 2-year follow-up.Cox proportional hazards regression was performed to identify independent predictors of MACE.Results:After propensity score matching,333 patients in the IVUS group were matched with 666 patients in the angiography group.Following matching,there were no significant differences in clinical characteristics and angiographic data between the two groups(all P>0.05).Regarding procedural data,IVUS group had a higher number of stents implanted,longer total stent length,larger average stent diameter,more frequent use of post-dilation balloons,larger post-dilation balloon diameter,and greater contrast agent usage(all P<0.05).MACE rate was significantly higher in the angiography group than that in the IVUS group(23.9%vs.18.0%,P=0.037).The difference was mainly attributed to a higher rate of cardiac death in the angiography group(16.1%vs.10.8%,P=0.013).Additionally,patients in angiography group had a significantly higher all-cause mortality rate compared to IVUS group(19.7%vs.13.8%,P=0.022).Multivariate cox regression analysis showed that IVUS guidance(HR=0.73,95%CI:0.55-0.99,P=0.042)was an independent protective factor,while hypertension(HR=1.60,95%CI:1.13-2.26,P=0.008),type 2 diabetes(HR=1.56,95%CI:1.19-2.05,P=0.001),acute coronary syndrome(HR=1.92,95%CI:1.12-3.30,P=0.018),and moderate to severe calcification(HR=1.55,95%CI:1.18-2.04,P=0.002)were independent risk factors for MACE at 2 years after DES implantation in CKD patients.Conclusions:Patients with CAD and CKD,IVUS-guided DES implantation is associated with a reduced risk of MACE and all-cause mortality at 2 years post-procedure compared to coronary angiography-guided implantation.

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 investigate the application of operation plan based on injury stress sequence in the treatment of degree Ⅲ and Ⅳ ankle fractures with Lange-Hansen classification of pronation and external rotation.Methods From May 2019 to May 2022,80 eligible patients of degree Ⅲand Ⅳ ankle fractures with Lange-Hansen classification were selected prospectively and randomely divided into 2 groups:40 patients in the study group were treated with surgical reduction according to the sequence of injury stress,and the other 40 patients in the control group were treated with traditional reduction sequence.The operative time,intraoperative blood loss,fracture healing time,full weight-bearing time,Ankle dorsoextension Angle,plantar flexion Angle,and American Orthopaedic Foot and Ankle Association were compared between the two groups Society,AOFAS)Ankle and posterior foot function scores and complications.Results The surgical time[(44.80±5.12)min vs.(47.61±6.42)min]and surgical bleeding volume[(43.81±9.02)ml vs.(51.50±12.01)ml]between the study group and the control group were statistically significant(all P＜0.05).Before surgery,there had no significant differences in ankle joint dorsiflexion angle,plantar flexion angle,AOFAS ankle and hind foot function score in 2 groups(all P＞0.05).The ankle dorsiflexion angle at 3 months after surgery[(16.84±1.77)° vs.(15.50±1.65)°],plantar flexion angle at 3 months after surgery[(45.64±4.10)° vs.(42.58±3.86)°]and at 6 months after surgery[(52.38±3.03)°vs.(50.64±3.74)°],the maximum walking distance score[(3.50±0.64)vs.(3.16±0.60)],ground walking score[(2.06±0.56)vs.(1.72±0.48)]and ankle joint anteroposterior activity score at 3 months after surgery[(4.96±1.38)vs.(4.16±1.07)]of the study group were all higher than those in the control group(all P＜0.05).There was no statistically significant difference in terms of fracture healing time,complete weight-bearing time,postoperative pain score,ankle joint function autonomous activity support score,abnormal gait score,maximum walking distance score at 6 months after surgery,ground walking score,ankle joint anterior and posterior activity score and surgical complications between 2 group(all P＞0.05).Conclusion According to the sequence of injury stress,surgical reduction for the treatment of grade Ⅲ and Ⅳ ankle fractures of Lange-Hansen classification of middle pronation and external rotation can shorten the operation time and promote the early recovery of joint function.

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.

Objectives:To assess the impact of intravascular ultrasound(IVUS)guidance for drug-eluting stent(DES)implantation on the long-term outcomes in coronary artery disease(CAD)patients with chronic kidney disease(CKD).Methods:A retrospective study was conducted on 1 800 CAD and CKD patients who underwent coronary DES implantation at Nanjing First Hospital from January 2018 to December 2022.Patients were divided into IVUS-guided group(IVUS group,n=333)and angiography-guided group(angiography group,n=1 467).Propensity score matching(PSM)was performed at a 1:2 ratio to adjust for differences in baseline clinical and angiographic characteristics between the two groups.Major adverse cardiovascular events(MACE),including cardiac death,target vessel myocardial infarction,and clinically driven target vessel revascularization,were evaluated over a 2-year follow-up.Cox proportional hazards regression was performed to identify independent predictors of MACE.Results:After propensity score matching,333 patients in the IVUS group were matched with 666 patients in the angiography group.Following matching,there were no significant differences in clinical characteristics and angiographic data between the two groups(all P>0.05).Regarding procedural data,IVUS group had a higher number of stents implanted,longer total stent length,larger average stent diameter,more frequent use of post-dilation balloons,larger post-dilation balloon diameter,and greater contrast agent usage(all P<0.05).MACE rate was significantly higher in the angiography group than that in the IVUS group(23.9%vs.18.0%,P=0.037).The difference was mainly attributed to a higher rate of cardiac death in the angiography group(16.1%vs.10.8%,P=0.013).Additionally,patients in angiography group had a significantly higher all-cause mortality rate compared to IVUS group(19.7%vs.13.8%,P=0.022).Multivariate cox regression analysis showed that IVUS guidance(HR=0.73,95%CI:0.55-0.99,P=0.042)was an independent protective factor,while hypertension(HR=1.60,95%CI:1.13-2.26,P=0.008),type 2 diabetes(HR=1.56,95%CI:1.19-2.05,P=0.001),acute coronary syndrome(HR=1.92,95%CI:1.12-3.30,P=0.018),and moderate to severe calcification(HR=1.55,95%CI:1.18-2.04,P=0.002)were independent risk factors for MACE at 2 years after DES implantation in CKD patients.Conclusions:Patients with CAD and CKD,IVUS-guided DES implantation is associated with a reduced risk of MACE and all-cause mortality at 2 years post-procedure compared to coronary angiography-guided implantation.

Objective To evaluate the efficacy and safety of polyethylene glycol electrolyte powder(PEG)combined with linaclotide(Lin)for bowel preparation in elderly constipated patients before colonoscopy.Methods In this prospective,randomized controlled trial,90 elderly patients with constipation undergoing colonoscopy were recruited at our hospital from June 2022 to December 2023.Participants were randomly assigned to three groups(n=30 each):PEG-3L alone,PEG-3L+Lin,and PEG-2L+Lin.Primary outcome was Boston Bowel Preparation Scale(BBPS)score and secondary outcomes included adverse event rates,colonoscopy completion rate,withdrawal time,and polyp detection rate.Statistical analysis was performed using independent t-tests and chi-square tests.Results The PEG-3L+Lin group achieved significantly higher BBPS scores than both PEG-3L alone and PEG-2L+Lin groups did(both P<0.001).The PEG-2L+Lin group also outperformed the PEG-3L alone group in cleansing efficacy(90.0%vs.76.7%,P=0.008).The PEG-2L+Lin group demonstrated the best tolerability and lowest adverse event rate,the PEG-3L group had the longest withdrawal time(P<0.05),but the three groups showed no significant difference in polyp detection rates.Conclusion PEG combined with linaclotide significantly improves bowel cleansing in elderly constipated patients.PEG-2L+Lin regimen provides optimal balance between efficacy,safety,and tolerability,making it a preferable choice for this population.