Background: Twenty-four-hour urinary free cortisol (UFC) measurement is the initial diagnostic test for Cushing’s syndrome (CS). We compared UFC determination by both direct and extraction immunoassays using Abbott Architect, Siemens Atellica Solution, and Beckman DxI800 with liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, we evaluated the value of 24-hr UFC measured by six methods for diagnosing CS. Methods: Residual 24-hr urine samples of 94 CS and 246 non-CS patients were collected.A laboratory-developed LC-MS/MS method was used as reference. UFC was measured by direct assays (D) using Abbott, Siemens, and Beckman platforms and by extraction assays (E) using Siemens and Beckman platforms. Method was compared using Passing–Bablok regression and Bland–Altman plot analyses. Cut-off values for the six assays and corresponding sensitivities and specificities were calculated by ROC analysis. Results: Abbott-D, Beckman-E, Siemens-E, and Siemens-D showed strong correlations with LC-MS/MS (Spearman coefficient r = 0.965, 0.922, 0.922, and 0.897, respectively), while Beckman-D showed weaker correlation (r = 0.755). All immunoassays showed proportionally positive bias. The areas under the curve were 0.975 for Abbott-D, 0.972 for LCMS/MS, 0.966 for Siemens-E, 0.948 for Siemens-D, 0.955 for Beckman-E, and 0.877 for Beckman-D. The cut-off values varied significantly (154.8–1,321.5 nmol/24 hrs). Assay sensitivity and specificity ranged from 76.1% to 93.2% and from 93.0% to 97.1%, respectively. Conclusions Commercially available immunoassays for measuring UFC show different levels of analytical consistency compared to LC-MS/MS. Abbott-D, Siemens-E, and Beckman-E have high diagnostic accuracy for CS.

Objective:Exploring the efficacy and safety of bridging blinatumomab (BiTE) in combination with chimeric antigen receptor T (CAR-T) cell therapy for the treatment of adult patients with acute B-cell lymphoblastic leukemia (B-ALL) .Methods:Clinical data from 36 adult B-ALL patients treated at the First Affiliated Hospital of Suzhou University from August 2018 to May 2023 were retrospectively analyzed. A total of 36 cases were included: 18 men and 18 women. The median age was 43.5 years (21-72 years). Moreover, 21 cases of Philadelphia chromosome-positive acute lymphoblastic leukemia were reported, and 16 of these cases were relapsed or refractory. Eighteen patients underwent blinatumomab bridging followed by CAR-T cell therapy, and 18 patients received CAR-T cell therapy. This study analyzed the efficacy and safety of treatment in two groups of patients.Results:In the BiTE bridge-to-CAR-T group, 16 patients achieved complete remission (CR) after BiTE immunotherapy, with a CR rate of 88.9%. One month after bridging CAR-T therapy, bone marrow examination showed a CR rate of 100.0%, and the minimal residual disease (MRD) negativity rate was higher than the nonbridging therapy group (94.4% vs. 61.1%, Fisher, P=0.041). The incidence of cytokine release syndrome and other adverse reactions in the BiTE bridge-to-CAR-T group was lower than that in the nonbridging therapy group (11.1% vs. 50.0%, Fisher, P=0.027). The follow-up reveals that 13 patients continued to maintain MRD negativity, and five patients experienced relapse 8.40 months (2.57-10.20 months) after treatment. Two of five patients with relapse achieved CR after receiving the second CAR-T cell therapy. In the nonbridging therapy group, 10 patients maintained continuous MRD negativity, 7 experienced relapse, and 6 died. The 1 year overall survival rate in the BiTE bridge-to-CAR-T group was higher than that in the nonbridging therapy group, with a statistically significant difference at the 0.1 level (88.9%±10.5% vs. 66.7%±10.9%, P=0.091) . Conclusion:BiTE bridging CAR-T cell therapy demonstrates excellent efficacy in adult B-ALL treatment, with a low recent recurrence rate and ongoing assessment of long-term efficacy during follow-up.

Objective To investigate the MRI characteristics of subventricular zone(SVZ)-associated isocitrate dehydrogenase(IDH)wild-type glioblastoma(GBM)and their correlations with Ki-67 expression and O6-methylguanine-DNA methyltransferase(MGMT)promoter methylation status.Methods A retrospective analysis was conducted on data of 78 patients with IDH wild-type GBM who underwent surgery and received pathological confirmation.Preoperative MRI contrast-enhanced T1 WI sequences were used to assess SVZ involvement,and postoperative molecular testing of tumor markers,including Ki-67 expression and MGMT methylation status,was utilized to categorize the patients accordingly.Results The SVZ involved(+)group(P＜0.001)and the MGMT(+)group(P=0.036)exhibited significantly larger tumor volumes.There were no significant differences between the groups in terms of gender,age,left/right hemispheric lateralization,or specific brain lobe distribution.There was no significant association between Ki-67 expression levels,MGMT methylation status,and SVZ involvement,respectively.Conclusion The SVZ(+)group and the MGMT(+)group demonstrates a wider range of tumor invasion.

The STAR tool was used to evaluate and analyze the science, transparency, and applicability of Chinese pathology guidelines and consensus published in medical journals in 2022. There were a total of 18 pathology guidelines and consensuses published in 2022, including 1 guideline and 17 consensuses. The results showed that the guideline score was 21.83 points, lower than the overall guideline average (43.4 points). Consensus ratings scored an average of 27.87 points, on par with the overall consensus level (28.3 points). Areas that scored above the overall level were "conflict of interest" and "working groups", while areas that scored below the overall level were "proposals", "funding", "evidence", "consensus approaches" and "accessibility". To sum up, the formulation of pathology guidelines and consensuses in 2022 is not standardized, and the evidence retrieval process, evidence evaluation methods and grading criteria for recommendations on clinical issues are not provided in the formulation process; the process and method for reaching consensus are not provided, the plan is lacking, and registration is not carried out. It is therefore suggested that guidelines/consensus makers in the field of pathology should attach importance to evidence-based medical evidence, strictly follow guideline formulation methods and processes, further improve the scientific, applicable and transparent guidelines/consensuses in the field, and better provide support for clinicians and patients.

Objective To investigate the mechanism by which lectin-like oxidized low density lipoprotein receptor-1(LOX-1)regulates hyperglycemic-induced myocardial fibroblast(CFs)fibrosis through the glycogen synthase kinase-3β(GSK-3β)/signal transducer and activator of transcription 3(STAT3)pathway.Methods CFs were isolated,cultured and identified.LOX-1 RNAi lentiviral vector was constructed and infected CFs.The experimental groups were as follows:Normal control(NC)group,High glucose(HG)group,LV-LOX-1,LV-Con group,Hypertonic(HPG)group.After LV-LOX-1 and LV-Con were infected with CFs,adding 25 mmol/L glucose to culture CFs for 24 h,they were denoted as HG+LV-LOX-1 group and HG+LV-Con group.Cells in HG+LV-LOX-1 group and HG+LV-Con group were treated with 10 μ mol/L SB216763 and 10 μ mol/L STATTIC for 24 h,respectively,and then they were recorded as HG+LV-LOX-1+SB216763 group,HG+LV-Con+SB216763 group,HG+LV-LOX-1+STATTIC group and HG+LV-Con+STATTIC group.CCK-8 was used to detect the activity of CFs,and the expression levels of mRAN and protein of LOX-1,collagen type I(COL-I),thioredoxin 5(TXNDC5),GSK-3β,STAT3,p-GSK-3β and p-STAT3 were detected by qRT-PCR and Western blot.Results CFs infected with LOX-1 RNAi lentiviral vector were obtained,which showed green under fluorescence microscopy.Compared with HG and HG+LV-Con groups,the mRNA expressions of LOX-1,COL-I and TXNDC5 were decreased in HG+LV-LOX-1 group(P<0.05).Compared with HG+LV-LOX-1 group,mRNA expressions of COL-I and TXNDC5 were decreased in HG+LV-LOX-1+SB216763 and HG+LV-LOX-1+STATTIC groups(P<0.05).Compared with HG and HG+LV-Con groups,p-GSK-3β protein expression was increased in HG+LV-LOX-1 group(P<0.05),while LOX-1,p-STAT3,COL-I,TXNDC5 protein expression was decreased in HG+LV-LOX-1 group(P<0.05).Compared with HG+LV-LOX-1 group,p-GSK-3β protein expression was increased in HG+LV-LOX-1+SB216763 group(P<0.05),while the protein expressions of p-STAT3,COL-I and TXNDC5 were decreased in HG+LV-LOX-1+SB216763 and HG+LV-LOX-1+STATTIC groups(P<0.05).Conclusion LOX-1,GSK-3β,STAT3,TXNDC5,and COL-I are involved in high glucose induced CFs fibrosis.LOX-1 promotes the expression of TXNDC5 and COL-I through GSK-3β/STAT3 pathway,and inhibition of LOX-1 can inhibit high glucose induced CFs fibrosis.

Deep phenotyping is a precise and comprehensive approach used for the precise analysis and comprehensive assessment of multi-system phenotypes of the patients. The approach uses symptoms, signs, various medical examination and laboratory results, and other relevant medical information. In the clinical diagnosis and medical research of rare bone diseases, deep phenotyping plays a pivotal role. The realization of precision medicine primarily comprises three key dimensions: deep phenotyping, stratified medicine, and targeted therapy. The deep phenotyping is the basis for the latter two. Deep phenotyping not only facilitates fine subtyping of diseases, but also allows for the in-depth understanding of genetic data. The use of deep phenotyping requires stand- ardized terminology and specific procedures. Moreover, deep phenotyping shows substantial potential using the application of artificial intelligence technology particularly when combining with multi-omics techniques.

Objective To explore the application value of automatic cytomorphological analyzer in the morphological analysis of white blood cells(WBC)in peripheral blood.Methods A total of 306 venous blood samples from inpatients and outpatients were randomly selected and prepared with automatic cytomorphological analyzer for WBC pre-classification.The differences between automatic cytomorphological analyzer counting,automatic blood cell analyzer counting and manual counting were compared,and the correlation between automatic cytomorphological analyzer and manual counting method was analyzed.Results Compared with the other two methods,the automatic cytomorphologi-cal analyzer was able to detect more types of WBC,especially abnormal cells.There were no signifi-cant differences between automatic cytomorphological analyzer and manual counting method for 6 ma-ture WBC types(band neutrophils,segmented neutrophils,lymphocytes,monocytes,eosinophils,and basophils),immature cells at different stages and atypical lymphocyte counts(P＞0.05).Re-sults of the 6 mature WBC types counted by the automatic cytomorphological analyzer and manual counting had favorable correlations(r＞0.8).Conclusion The automatic cytomorphological analyzer can classify more types of WBC,provide WBC counting results that are highly consistent with manual microscopy,and the counting results of the two methods have a good correlation.

Objective To explore the application value of automatic cytomorphological analyzer in the morphological analysis of white blood cells(WBC)in peripheral blood.Methods A total of 306 venous blood samples from inpatients and outpatients were randomly selected and prepared with automatic cytomorphological analyzer for WBC pre-classification.The differences between automatic cytomorphological analyzer counting,automatic blood cell analyzer counting and manual counting were compared,and the correlation between automatic cytomorphological analyzer and manual counting method was analyzed.Results Compared with the other two methods,the automatic cytomorphologi-cal analyzer was able to detect more types of WBC,especially abnormal cells.There were no signifi-cant differences between automatic cytomorphological analyzer and manual counting method for 6 ma-ture WBC types(band neutrophils,segmented neutrophils,lymphocytes,monocytes,eosinophils,and basophils),immature cells at different stages and atypical lymphocyte counts(P＞0.05).Re-sults of the 6 mature WBC types counted by the automatic cytomorphological analyzer and manual counting had favorable correlations(r＞0.8).Conclusion The automatic cytomorphological analyzer can classify more types of WBC,provide WBC counting results that are highly consistent with manual microscopy,and the counting results of the two methods have a good correlation.

Objective:To investigate the clinical value of virtual monoenergetic images (VMI) and electron density map (EDM) derived from the dual-layer spectral detector CT (DLCT) in the differential diagnosis of benign and malignant pulmonary ground glass nodules (GGN).Methods:From July 2019 to August 2020, a total of 65 patients with lung GGN (27 benign GGNs and 38 malignant GGNs) confirmed by pathology were retrospectively enrolled in Gulou Clinical Medical College of Nanjing Medical University. All the patients underwent DLCT plain scanning within two weeks before the surgery. The conventional 120 kVp polyenergetic image (PI), EDM and 40-80 keV VMI were reconstructed. The differences of CT and electron density (ED) values between benign and malignant lesions on different images were compared by Mann-Whitney U test. Independent t-test was used to compare the lesion size and χ 2 test was used to analyze the CT features (including lesion location, shape, edge, internal structure, adjacent structure, nodule type) between benign and malignant lesions. Receiver operating characteristic (ROC) curve was used to analyze the efficacy of different energy spectrum quantitative parameters in the differential diagnosis of benign and malignant GGN. The statistically significant CT signs and energy spectrum quantitative parameters were analyzed by logistic regression analysis to find out the independent risk factors of malignant GGN, and then ROC curve analysis was performed for each independent risk factor alone or in combination. Results:There were significant differences in lesion shape, spiculation, lobulation, location and size between benign and malignant groups ( P<0.05). The CT value of pulmonary GGN in PI, 40-80 keV VMI and the ED value in EDM were statistically different between benign and malignant lesions ( P<0.05). The area under ROC curve (AUC) were 0.680, 0.682, 0.683, 0.686, 0.694, 0.676 and 0.722, respectively, among which the ED value had the highest AUC. Binary logistic regression analysis was carried out with GGN shape, spiculation, lobulation, location, size, ED value and CT value in PI, 40-80 keV VMI as independent variables, and malignant GGN as dependent variables. The results showed that ED value (OR=1.045, 95%CI 1.001-1.090, P=0.044), lesion size (OR=1.582, 95%CI 1.159-2.158, P=0.004), spiculation sign (OR=11.352, 95%CI 2.379-54.172, P=0.002) were independent risk factors for malignant GGN. ROC curve analysis showed the AUC of ED value, lesion size, spiculation sign and combination of the three for differential diagnosis of benign and malignant GGN were 0.722, 0.772, 0.698 and 0.885. The AUC for the combined parameters was the largest, with sensitivity of 92.1% and specificity of 74.1%. Conclusion:The diagnostic efficacy of EDM is higher than that of other VMI in the differential diagnosis of pulmonary GGN by DLCT images; The efficacy is further improved when EDM is combined with lesion size and spiculation sign for comprehensive diagnosis.

Objective:To explore the clinical efficacy of consolidation chemotherapy combined with microtransplantation in the treatment of elderly patients with acute myeloid leukemia (AML).Methods:The clinical data of 45 elderly patients with AML in Suzhou Yongding Hospital from January 2016 to December 2018 were retrospectively analyzed. After 1 or 2 courses of induced chemotherapy, the patients achieved complete response (CR). Among them, 20 patients were given consolidation chemotherapy (single group), and 25 patients were given consolidation chemotherapy combined with microtransplantation (combined group). After 3 courses of treatment, the minimal residual disease (MRD), quantitative expression of WT1 gene and adverse reactions were compared between 2 groups; the quality of life before treatment and after treatment was evaluated by quality of life-BREF (QOL-BREF). The patients were followed up to November 1, 2020, and the median follow-up was 30 months. The overall survival and progress-free survival (PFS) were recorded.Results:The MRD negative rate and WT1 gene negative rate after treatment in combined group were significantly higher than those in single group: 60.00% (15/25) vs. 25.00% (5/20) and 52.00% (13/25) vs. 20.00% (4/20), and there were statistical differences ( P<0.05). The recovery times of neutrophils and platelets in combined group were significantly shorter than those in single group: 10 (8, 12) d vs. 16 (13, 20) d and 14 (11, 17) d vs. 24 (19, 30) d, and there were statistical differences ( Z = 3.152 and 4.285, P<0.05). No adverse reactions such as liver and kidney abnormalities or gastrointestinal reactions occurred in 2 groups; and no specific graft versus host disease (GVHD) occurred in the combined group. The each item scores of QOL-BREF after treatment in combined group were significantly higher than those in single group, and there were statistical differences ( P<0.01). The 2-year overall survival rate and PFS rate in combined group were significantly higher than those in single group: 60.00% (15/25) vs. 35.00% (7/20) and 52.00% (13/25) vs. 25.00% (5/20), and there were statistical differences ( χ2 = 4.235 and 4.742, P = 0.040 and 0.029). Conclusions:Consolidation chemotherapy combined with microtransplantation is effective and safe in the treatment of elderly patients with AML. It can significantly improve the quality of life, and improve the overall survival rate and PFS rate.