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 evaluate the outcomes of intensive conservative treatment compared to conventional conservative treatment in patients with acute aortic syndrome(AAS).Methods:The study prospectively enrolled consecutive patients with AAS who were admitted to Beijing Anzhen Hospital, affiliated with Capital Medical University, and Beijing Dawanglu Emergency Rescue Hospital from January 2024 to December 2024. These patients with surgical contraindications or refused surgery for various reasons opted for conservative treatment. A total of 282 patients were included, and 15 patients with missing data or those who died without any treatment were excluded. Finally, 267 patients were enrolled, of whom 94 received intensive conservative treatment, and 173 received conventional conservative treatment, the inverse probability of treatment weighting (IPTW) was used to reduce the influence of confoundings. After adjusting of baseline datas via IPTW, the survival outcomes of the two groups were compared at 14 days, 30 days, and at the end of follow-up.Results:The results showed significant differences in acute phase survival rates between the enhanced conservative treatment group and the conventional conservative treatment group at 14 days(82.40%vs.53.20%, P<0.0001). Significant survival differences were also observed at 30 days and at 276-day mid-term follow-up (96.29% vs.51.60%, P<0.0001; 78.50% vs.48.50%, P<0.0001). In the subgroup analysis, for type A aortic dissection, the enhanced conservative treatment group had higher survival rates compared to the conventional conservative treatment group at 14, 30 and 276 days (63.46% vs.41.35%, P<0.05; 52.17% vs.37.90%, P<0.05; 50.00% vs. 31.97%, P<0.05). However, for type B aortic dissection, although the enhanced conservative treatment group had higher survival rates than the conventional conservative treatment group, no statistically significant differences were observed (96.29% vs. 80.00%, P=0.054; 95.65% vs.78.37%, P=0.067; 94.12% vs.74.20%, P=0.088). Conclusion:For patients diagnosed with AAS are forced to choose conservative treatment if emergency surgery is not possible in the first place, intensive conservative treatment strategies can significantly reduce the mortality in the acute phase compared with conventional conservative treatment. Mid-term follow-up, intensive conservative treatment still has a significant survival advantage.

Objective:To understand the experience of inpatients′ needs on medical nursing assistants, and explore the characteristics and population differences, in order to provide a reference for optimizing the inpatient companion management system.Methods:The descriptive qualitative study was adopted for semi-structured in-depth interviews of 19 inpatients hospitalized in Zhongshan Hospital Fudan University from December 2023 to January 2024 based on purposive sampling. Information on personal characteristics, status and experience of dependence on medical nursing assistants was collected. Colaizzi seven-step analysis method was used to construct the fact labels and model dimensions. Manually extract the characteristics of inpatients′ needs on medical nursing assistants, and construct a user persona to complete the visualization.Results:A total of 19 inpatients′ were interviewed, including 12 males and 7 females with an average age of (63.16 ± 11.56) years. Four dimensions were extracted including cognitive characteristics, health status characteristics, family support characteristics, and psychological characteristics. Combined with personal traits and experience, three types of typical user persona were constructed including independent type, mutual assistance and cooperation type, complete dependence type.Conclusions:Group heterogeneity existed in the demand of inpatients on medical nursing assistants. According to the user persona, the manager should formulate personalized and precise caregiver management strategies.

Objective To explore the characteristics and influencing factors of complicated intra-abdominal infection in ICU patients after abdominal surgery.Methods A retrospective study was performed on ICU patients(n=92,observation group)developing complicated intra-abdominal infection after abdominal surgery versus patients(n=104,control group)without complicated intra-abdominal infection after abdominal surgery in our hospital from January 2020 to December 2023.The characteristics of patients who developed complicated intra-abdominal infections were analyzed and the relevant influencing factors were identified using multivariate Logistic regression analysis.Results A total of 124 strains of pathogenic bacteria were isolated and identified in 92 infected patients,with 64.52％(80/124)of Gram-negative strains comprising mainly 29.03％(36/124)of Escherichia coli,25.81％(32/124)of Gram-positive strains comprising mainly 12.90％(16/124)of Enterococcus faecalis,and 9.68％(12/124)of fungi comprising mainly 6.45％(8/124)of Candida albicans.Multivariate Logistic regression results showed that preoperative underlying disease,surgical methods,duration of surgery,invasive procedures,antibiotic use,and length of ICU stay,enteral nutrition were the influencing factors of complicated intra-abdominal infection in ICU patients after abdominal surgery(P＜0.0 5).Conclusion Patients with complicated intra-abdominal infection are infected with a variety of pathogenic bacteria,predominantly Gram-negative.The patient's preoperative underlying disease,surgical approach,duration of surgery,invasive procedures,use of antibiotics,length of ICU stay,and early enteral nutrition all affect the risk of complicated intra-abdominal infection in ICU patients after abdominal surgery.

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.

Objective:To explore the influencing factors for pathological upgrading in prostate cancer patients with a Gleason score of 3 + 3 undergoing targeted biopsy，and to establish a nomogram prediction model.Methods:A retrospective analysis was conducted on 191 patients with localized prostate cancer diagnosed with a Gleason score of 3 + 3 through targeted biopsies at the First Affiliated Hospital of Nanjing Medical University from January 2020 to June 2024. The age of the patients was 67（61，73）years，with prostate-specific antigen（PSA）level of 7.44（5.53，10.19）ng/ml，prostate volume of 35.64（26.59，48.97）ml，and PSA density（PSAD）of 0.20（0.14，0.31）ng/ml 2. Among them，61 cases（31.94%）had a Prostate Imaging Reporting and Data System（PI-RADS）score of 3，104 cases（54.45%）had a score of 4，and 26 cases（13.61%）had a score of 5. The diameter of the main lesion was 10.75（7.86，14.00）mm. The lesions were located in the peripheral zone in 78 cases（40.84%），the transition zone in 99 cases（51.83%），and the anterior fibromuscular stroma in 14 cases（7.33%）. The lesions were found at the apex in 56 cases（29.32%），in the body in 120 cases（62.83%），and at the base in 15 cases（7.85%）. MRI revealed only one lesion with a PI-RADS score ≥ 3 in 131 cases，two suspected lesions in 43 cases，three suspected lesions in 12 cases，and four suspected lesions in 5 cases. Systematic biopsy was positive in 121 cases（63.4%）and negative in 70 cases（36.6%）. The lesions were confined to the left lobe in 63 cases（32.98%），right lobe in 68 cases（35.60%），and involved both lobes in 60 cases（31.41%）. The interval between biopsy and surgery was 9.0（7.0，14.0）days. Univariate analyses were performed using Mann-Whitney U tests or χ2 tests，and multivariate logistic regression was used to identify independent predictors of pathological upgrading. A nomogram model was constructed based on these independent predictors. The model’s discriminative ability was assessed using the area under the receiver operating characteristic（ROC）curve（AUC），and internal validation of the model’s consistency was conducted using the bootstrap resampling method. Decision curve analysis（DCA）was performed to assess clinical utility. Results:Among the 191 cases，60（31.4%）had no pathological upgrading after surgery，while 131（68.6%）showed upgrading. Univariate analysis showed that the maximum diameter of the main lesion［9.0（6.0，13.2）mm vs. 11.0（8.4，14.0）mm］，number of suspicious lesions on MRI［1.0（1.0，1.0）vs. 1.0（1.0，2.0）］，number of positive systematic biopsy cores［1.0（0，2.0）vs. 1.0（0，3.0）］，percentage of positive systematic biopsy cores［0.08（0，0.17）vs. 0.12（0，0.25）］，number of positive targeted biopsy cores［2.0（1.0，3.0）vs. 3.0（1.0，4.0）］，percentage of positive targeted biopsy cores［0.37（0.24，0.75）vs. 0.50（0.38，0.85）］，level of the index lesion，location of the index lesion，and PI-RADS score were associated with pathological upgrading（ P < 0.05）. Multivariate logistic regression analysis showed that PI-RADS score 4（ OR = 5.88，95% CI 2.41 - 14.35），number of suspicious lesions on MRI（ OR = 4.15，95% CI 1.88 - 9.17），location of the index lesion in the transition zone（ OR = 6.86，95% CI 2.81 - 16.73），and percentage of positive targeted biopsy cores（ OR = 4.37，95% CI 1.38 - 14.90）were independent risk factors for pathological upgrading（ P < 0.05）. The nomogram model constructed using these predictors had an AUC of 0.845. Internal validation using the Bootstrap method yielded an AUC value of 0.812，indicating high predictive accuracy of the model. The calibration curve indicated good calibration. Decision curve analysis showed that the threshold range for net benefit in the model was between 12% - 100%. Conclusions:The PI-RADS score 4，the number of lesions with PI-RADS ≥ 3，the location of the main lesion in the transition zone，and the percentage of positive needles in targeted biopsy are independent risk factors for pathological upgrading from Gleason score 3 + 3. The nomogram model constructed from these factors demonstrates good predictive performance and provides a reference for clinical decision-making.

Objective:To evaluate the outcomes of intensive conservative treatment compared to conventional conservative treatment in patients with acute aortic syndrome(AAS).Methods:The study prospectively enrolled consecutive patients with AAS who were admitted to Beijing Anzhen Hospital, affiliated with Capital Medical University, and Beijing Dawanglu Emergency Rescue Hospital from January 2024 to December 2024. These patients with surgical contraindications or refused surgery for various reasons opted for conservative treatment. A total of 282 patients were included, and 15 patients with missing data or those who died without any treatment were excluded. Finally, 267 patients were enrolled, of whom 94 received intensive conservative treatment, and 173 received conventional conservative treatment, the inverse probability of treatment weighting (IPTW) was used to reduce the influence of confoundings. After adjusting of baseline datas via IPTW, the survival outcomes of the two groups were compared at 14 days, 30 days, and at the end of follow-up.Results:The results showed significant differences in acute phase survival rates between the enhanced conservative treatment group and the conventional conservative treatment group at 14 days(82.40%vs.53.20%, P<0.0001). Significant survival differences were also observed at 30 days and at 276-day mid-term follow-up (96.29% vs.51.60%, P<0.0001; 78.50% vs.48.50%, P<0.0001). In the subgroup analysis, for type A aortic dissection, the enhanced conservative treatment group had higher survival rates compared to the conventional conservative treatment group at 14, 30 and 276 days (63.46% vs.41.35%, P<0.05; 52.17% vs.37.90%, P<0.05; 50.00% vs. 31.97%, P<0.05). However, for type B aortic dissection, although the enhanced conservative treatment group had higher survival rates than the conventional conservative treatment group, no statistically significant differences were observed (96.29% vs. 80.00%, P=0.054; 95.65% vs.78.37%, P=0.067; 94.12% vs.74.20%, P=0.088). Conclusion:For patients diagnosed with AAS are forced to choose conservative treatment if emergency surgery is not possible in the first place, intensive conservative treatment strategies can significantly reduce the mortality in the acute phase compared with conventional conservative treatment. Mid-term follow-up, intensive conservative treatment still has a significant survival advantage.

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 mechanism of Tongdu Xingshen acupuncture,the clinical efficacy,systemic inflammatory response,blood-brain barrier and intestinal flora in patients with cognitive impairment after ischemic stroke(IS)were studied.Methods Thirty patients(3 cases shedding)with cognitive impairment after IS were included as the disease group,including patients before treatment as the disease group,patients after Tongdu Xingshen acupuncture treatment as the electroacupuncture group,and 30 healthy controls(3 cases shedding)were included as the healthy group.In the electroacupuncture group,on the basis of the basic treatment,Tongdu Xingshen acupuncture was applied,which was 30 min each time,once a day for 14 days.The MMSE,MoCA and MBI scores of the three groups were observed.The fecal and serum samples from all study subjects were collected,and 16S rDNA sequencing technology and ELISA were used to detect the changes of proinflammatory factors IL-6,IL-1β,TNF-α and S100β in serum in intestinal flora and feces.Results Compared with the healthy group,the MMSE,MoCA,and MBI score of patients in the disease group decreased significantly(P<0.05),serum proinflammatory factors and S100β protein content increased significantly(P<0.05),and the Shannon index(P<0.01)and Simpson index(P<0.001)increased significantly.Compared with the disease group,the MMSE,MoCA,and MBI score of the EA group increased significantly(P<0.05),the serum levels of proinflammatory factors and S100β decreased significantly(P<0.05),Shannon index and Simpson index decreased(P>0.05).The dominant bacterial flora in the healthy group mainly included Bacteroides,Bifidobacterium,Bacteroides,Faecalibacterium,Bifidobacteriaceae,Ruminococcaceae,and Bacteroides and other beneficial bacteria(P<0.05).The dominant flora in the disease group included Proteobacteria,Enterobacteriaceae,Escherichia,Klebsiella and other opportunistic bacteria(P<0.05),while the dominant flora in the EA group was consistent with the healthy group,the relative abundance of beneficial bacteria increased significantly(P<0.05),and the relative abundance of opportunistic bacteria decreased significantly(P<0.05).Spearman correlation analysis found that beneficial bacteria were positively correlated with clinical efficacy related indicators,but with serum proinflammatory factors and the content of S100β was negatively correlated.Conclusion Tongdu Xingshen acupuncture can regulate the diversity of intestinal flora to increase the abundance of Bacteroides,Bifidobacterium,Faecalibacterium,and other beneficial bacteria,regulate the intestinal microecological balance,Thereby regulating systemic inflammation and blood-brain barrier function,which plays a role in improving cognitive function.

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.