Objective:To explore the application value of the artificial intelligence (AI) morphological assisted analysis system in the pre-classification of blood cells in patients with acute myeloid leukemia, myelodysplasia-related (AML-MR).Methods:A retrospective analysis was conducted on the bone marrow and peripheral blood cell morphology of patients initially diagnosed with AML-MR at Taizhou Hospital in Zhejiang Province from September 1, 2022, to December 31, 2023. A total of 44 patients, including 25 males and 19 females, with a median age of 71 (63.5, 75.3) years. Bone marrow and peripheral blood morphology were examined using the Morphogo cell morphology assisted analysis system, with the artificial classification results serving as the gold standard. A confusion matrix was constructed to evaluate the precision, sensitivity, and specificity of the AI system in identifying various cell types in bone marrow and peripheral blood for AML-MR diagnosis. The impact of dysplastic hematopoiesis on AI pre-classification was analyzed by comparing AI and manual classification results.Results:The AI system completed the pre-classification of 44 bone marrow smears and 42 corresponding peripheral blood smears from AML-MR patients. For bone marrow smears, the precision, sensitivity, and specificity of AI in pre-classifying blast cells were 85.78%, 91.01%, and 94.58%, respectively. For peripheral blood smears, these values were 87.11%, 87.05%, and 98.29%, respectively. The precision and sensitivity of AI in pre-classifying promyelocytes were 54.26% and 46.93%, respectively, while for monocytes, they were 58.16% and 68.34%, both lower than those for blast cells. The precision and sensitivity of AI in identifying myelocytes and metamyelocytes also decreased (77.47%, 66.25% and 81.91%, 63.29%, respectively). The precision and sensitivity of AI in pre-classifying erythroblasts/proerythroblasts (67.71%, 69.89%) were lower than those for polychromatic and orthochromatic normoblasts (83.43%, 85.53% and 92.97%, 86.96%, respectively). The confusion matrix and comparative analysis of AI and manual classification indicated that the decline in AI pre-classification precision and sensitivity was due to frequent misclassification between promonocytes and monocytes, as well as between monocytes and promyelocytes. Additionally, this decline is associated with dysplasia. However, the impact of dysplasia on the AI pre-classification of mature-stage granulocytes was minimal.Conclusion:The AI system demonstrated high precision, sensitivity, and specificity in pre-classifying blast cells in bone marrow and peripheral blood smears from AML-MR patients. The AI-assisted morphological analysis system can be effectively utilized for the pre-classification of blood cells in AML-MR patients.

Objective:To explore the immunophenotypic differences between acute promyelocytic leukemia (APL) and APL-like NPM1 mutant acute myeloid leukemia (NPM1mutAML) using flow cytometry, and to investigate early diagnostic markers for differentiating APL from NPM1mutAML.Methods:A retrospective study was conducted on 72 cases of APL diagnosed at Taizhou Hospital, affiliated with Wenzhou Medical University, from February 2nd, 2018 to December 16th, 2023, including 42 male and 30 female patients with a median age of 42 (32, 57) years old. Based on morphology, 51 cases were classified as the coarse-granular type and 21 cases as the fine-granular type. Additionally, 45 cases of NPM1mutAML, comprising 20 male and 25 female patients with a median age of 58 (47, 65) years old, were included. Of these, 12 cases were classified as the coarse-granular type and 33 as the fine-granular type. Immunophenotypic analysis was performed using multiparameter flow cytometry, and all patients underwent cytogenetic analysis for chromosome karyotyping. FISH analysis was used for detecting the PML-RARα fusion gene in APL cases, and sequencing was used for identifying NPM1 mutations in NPM1mutAML patients. The antigen expression parameters (expression rate, median fluorescence intensity [MdFI], and coefficient of variation [ CV]) were analyzed using principal component analysis (PCA). The antigen expression rates were compared using the Wilcoxon rank-sum test, and the positive rates of antigens were compared using the Chi-square test. Sensitivity and specificity for diagnosis by the some antigens were evaluated using ROC curve analysis. Results:The immunophenotypic analysis revealed that the expression rates of CD123, CD64, CD13, and CD9 were significantly higher in APL compared to NPM1mutAML ( Z values of-6.72, -6.29, -5.63, -7.67, P<0.01). In the coarse-granular type, the expression rates of CD123 and CD9 in APL were also significantly higher than those in NPM1mutAML ( P<0.01). In the fine-granular type, the expression levels of CD123, CD13, CD64, and CD9 were significantly higher in APL than in NPM1mutAML ( P<0.01). ROC curve analysis showed that in the fine-granular type, the areas under the curve (AUC) for CD64, CD13, CD123, and CD9 in diagnosing APL and NPM1mutAML were 0.96, 0.89, 0.86, and 0.89, respectively ( P<0.01). In the coarse-granular type, the AUC for CD64 and CD13 were 0.49 and 0.51 ( P>0.05), while the AUC for CD123 and CD9 were 0.96 and 0.96 ( P<0.01). Principal component analysis (PCA) of antigen expression (expression rate, MdFI, CV) showed complete separation of the APL and NPM1mutAML groups. Conclusion:APL and APL-like NPM1mutAML patients exhibit distinct antigen expression profiles. Specifically, a combined detection of CD64, CD13, CD123, and CD9 can help to rapidly differentiate APL from APL-like NPM1mutAML at initial diagnosis.

Objective:To explore the application value of the artificial intelligence (AI) morphological assisted analysis system in the pre-classification of blood cells in patients with acute myeloid leukemia, myelodysplasia-related (AML-MR).Methods:A retrospective analysis was conducted on the bone marrow and peripheral blood cell morphology of patients initially diagnosed with AML-MR at Taizhou Hospital in Zhejiang Province from September 1, 2022, to December 31, 2023. A total of 44 patients, including 25 males and 19 females, with a median age of 71 (63.5, 75.3) years. Bone marrow and peripheral blood morphology were examined using the Morphogo cell morphology assisted analysis system, with the artificial classification results serving as the gold standard. A confusion matrix was constructed to evaluate the precision, sensitivity, and specificity of the AI system in identifying various cell types in bone marrow and peripheral blood for AML-MR diagnosis. The impact of dysplastic hematopoiesis on AI pre-classification was analyzed by comparing AI and manual classification results.Results:The AI system completed the pre-classification of 44 bone marrow smears and 42 corresponding peripheral blood smears from AML-MR patients. For bone marrow smears, the precision, sensitivity, and specificity of AI in pre-classifying blast cells were 85.78%, 91.01%, and 94.58%, respectively. For peripheral blood smears, these values were 87.11%, 87.05%, and 98.29%, respectively. The precision and sensitivity of AI in pre-classifying promyelocytes were 54.26% and 46.93%, respectively, while for monocytes, they were 58.16% and 68.34%, both lower than those for blast cells. The precision and sensitivity of AI in identifying myelocytes and metamyelocytes also decreased (77.47%, 66.25% and 81.91%, 63.29%, respectively). The precision and sensitivity of AI in pre-classifying erythroblasts/proerythroblasts (67.71%, 69.89%) were lower than those for polychromatic and orthochromatic normoblasts (83.43%, 85.53% and 92.97%, 86.96%, respectively). The confusion matrix and comparative analysis of AI and manual classification indicated that the decline in AI pre-classification precision and sensitivity was due to frequent misclassification between promonocytes and monocytes, as well as between monocytes and promyelocytes. Additionally, this decline is associated with dysplasia. However, the impact of dysplasia on the AI pre-classification of mature-stage granulocytes was minimal.Conclusion:The AI system demonstrated high precision, sensitivity, and specificity in pre-classifying blast cells in bone marrow and peripheral blood smears from AML-MR patients. The AI-assisted morphological analysis system can be effectively utilized for the pre-classification of blood cells in AML-MR patients.

Objective:To explore the immunophenotypic differences between acute promyelocytic leukemia (APL) and APL-like NPM1 mutant acute myeloid leukemia (NPM1mutAML) using flow cytometry, and to investigate early diagnostic markers for differentiating APL from NPM1mutAML.Methods:A retrospective study was conducted on 72 cases of APL diagnosed at Taizhou Hospital, affiliated with Wenzhou Medical University, from February 2nd, 2018 to December 16th, 2023, including 42 male and 30 female patients with a median age of 42 (32, 57) years old. Based on morphology, 51 cases were classified as the coarse-granular type and 21 cases as the fine-granular type. Additionally, 45 cases of NPM1mutAML, comprising 20 male and 25 female patients with a median age of 58 (47, 65) years old, were included. Of these, 12 cases were classified as the coarse-granular type and 33 as the fine-granular type. Immunophenotypic analysis was performed using multiparameter flow cytometry, and all patients underwent cytogenetic analysis for chromosome karyotyping. FISH analysis was used for detecting the PML-RARα fusion gene in APL cases, and sequencing was used for identifying NPM1 mutations in NPM1mutAML patients. The antigen expression parameters (expression rate, median fluorescence intensity [MdFI], and coefficient of variation [ CV]) were analyzed using principal component analysis (PCA). The antigen expression rates were compared using the Wilcoxon rank-sum test, and the positive rates of antigens were compared using the Chi-square test. Sensitivity and specificity for diagnosis by the some antigens were evaluated using ROC curve analysis. Results:The immunophenotypic analysis revealed that the expression rates of CD123, CD64, CD13, and CD9 were significantly higher in APL compared to NPM1mutAML ( Z values of-6.72, -6.29, -5.63, -7.67, P<0.01). In the coarse-granular type, the expression rates of CD123 and CD9 in APL were also significantly higher than those in NPM1mutAML ( P<0.01). In the fine-granular type, the expression levels of CD123, CD13, CD64, and CD9 were significantly higher in APL than in NPM1mutAML ( P<0.01). ROC curve analysis showed that in the fine-granular type, the areas under the curve (AUC) for CD64, CD13, CD123, and CD9 in diagnosing APL and NPM1mutAML were 0.96, 0.89, 0.86, and 0.89, respectively ( P<0.01). In the coarse-granular type, the AUC for CD64 and CD13 were 0.49 and 0.51 ( P>0.05), while the AUC for CD123 and CD9 were 0.96 and 0.96 ( P<0.01). Principal component analysis (PCA) of antigen expression (expression rate, MdFI, CV) showed complete separation of the APL and NPM1mutAML groups. Conclusion:APL and APL-like NPM1mutAML patients exhibit distinct antigen expression profiles. Specifically, a combined detection of CD64, CD13, CD123, and CD9 can help to rapidly differentiate APL from APL-like NPM1mutAML at initial diagnosis.

Objective: To understand the epidemiological characteristics of pulmonary tuberculosis (PTB) in Shaanxi Province from 2016 to 2021, so as to provide the basis for improving PTB prevention and control strategies. Methods: The incidence data of PTB cases in Shaanxi Province were collected from the Tuberculosis Information Management System of the Chinese Disease Prevention and Control Information System from 2016 to 2021. The temporal, geographical, and demographic distribution characteristics of PTB were described. Results: A total of 121 280 PTB cases were reported in Shaanxi Province from 2016 to 2021, with the reported incidence rate decreasing from 56.30/105 in 2016 to 40.98/105 in 2021 (P<0.05). The etiological positivity rate increased from 16.61% in 2016 to 54.37% in 2021 (P<0.05). The average annual number of reported cases peaked in March (1 984.67 cases, accounting for 9.81%), followed by January (1 885.50 cases, accounting for 9.33%), and hit its lowest in October (1 458.50 cases, accounting for 7.22%). The incidence of PTB was relatively high in northern Shaanxi and southern Shaanxi, at 67.99/105 and 64.05/105, respectively, while it was relatively low in Guanzhong Region, at 44.83/105. The incidence rates of Ankang City (85.81/105), Fugu County (73.92/105), and Yulin City (72.20/105) ranked the top three. There were 80 111 male cases and 41 169 female cases of PTB, with a male-to-female ratio of 1.95︰1. The reported incidence rate of males (67.38/105) was significantly higher than that of females (36.61/105), and the difference was statistically significant (P<0.05). Among males, the highest average reported incidence rate was found in the 80 to <85 age group (179.46/105), while among females, the highest was in the 75 to <80 age group (104.38/105). The main occupations were farmers, accounting for 68.87% (83 526 cases). Conclusions The reported incidence rate of PTB in Shaanxi Province showed an overall downward trend from 2016 to 2021, with a high incidence in winter and spring. The epidemic in southern Shaanxi and northern Shaanxi is more severe than in Guanzhong Region of Shaanxi Province. The incidence of PTB is predominantly found among males, the elderly, and farmers.

Objective:To explore the expression levels and significance of programmed death factor 1 (PD-1)/programmed death factor ligand 1 (PDL-1) in T cells, regulatory T cells (Tregs), and regulatory B cells (Bregs) in patients with unexplained recurrent spontaneous abortion (URSA).Methods:Forty-two URSA patients (as patient group), 34 healthy pregnant women (as normal pregnancy group) and 30 unpregnant healthy examination patients (as control group) were collected for retrospective analysis,all study subjects were from patients who were treated in Taizhou Hospital affiliated to Wenzhou Medical University from February 2020 to February 2022. Flow cytometry was used to detect the expression level of PD-1/PDL-1 in Treg cells, Breg cells and [T cells, B cells and natural killer cells (TBNK)] lymphocyte subsets, as well as the expression level of serum Th1 (IFN-γ, TNF-α)/Th2 (IL-4, IL-6, IL-10)/Th17 (IL-17) cytokine expression. The patients were treated with lymphocyte immunotherapy, the changes of PD-1/PDL-1 levels were detected at the end of treatment, and the pregnancy outcome was recorded during follow-up. Comparisons between multiple groups were performed by ANOVA, comparisons before and after treatment in URSA patients were performed by paired T-test, and correlation of each test index by bivariate correlation analysis.Results:The expression levels of PD-1/PDL-1 in Treg, Breg cells and T lymphocyte subsets in peripheral blood of patients with URSA was significantly lower than that of healthy pregnant women(all P<0.01). The expression level of PD-1/PDL-1 in CD4+T cells was negatively correlated with the expression of serum Th1 cytokines Interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) (The r values were -0.44, -0.85, -0.33, and -0.94, respectively, all P<0.01). The expression level of PD-1/PDL-1 in CD4+T cells was positively correlated with the expression of serum Th2 cytokines interleukin 4(IL-4), interleukin 6(IL-6) and interleukin 10(IL-10)(The r values were 0.55, 0.47, 0.41, 0.33, 0.46, and 0.69, respectively,all P<0.01). The proportion of Breg cells and Treg cells were positively correlated with the level of serum IL-10 expression(The r values were 0.97, and 0.95 respectively, all P<0.01). The proportion of Treg cells was negatively correlated with the expression of IL-17( r=?0.95, P<0.01). The expression of PD-1/PDL-1 in Breg cells and Treg cells was positively correlated with the expression of serum IL-10(The r values were 0.95, 0.36, 0.96, and 0.95, respectively, all P<0.01). There was a negative correlation between serum IL-10 expression level and IL-17 expression level( r=?0.58, P<0.01). After URSA treatment, pregnancy was successful in 23 cases and failed in 19 cases. The expression of PD-1/PD-L1 on CD4, CD8, Tregs cells and Bregs cells in USRA treatment group was significantly higher than that before treatment( P<0.01), but there was no significant change in treatment failure group( P>0.05). Conclusion:The low expression of PD-1/PD-L1 in Treg cells, Breg cells and T cell subsets in peripheral blood of patients with URSA results in the immune imbalance of Th1/Th2 and Tregs/Th17, and the damage of maternal-fetal immune tolerance leads to pregnancy failure, which may be a potential therapeutic target.

METTL3 and METTL14 are two components that form the core heterodimer of the main RNA m6A methyltransferase complex (MTC) that installs m6A. Surprisingly, depletion of METTL3 or METTL14 displayed distinct effects on stemness maintenance of mouse embryonic stem cell (mESC). While comparable global hypo-methylation in RNA m6A was observed in Mettl3 or Mettl14 knockout mESCs, respectively. Mettl14 knockout led to a globally decreased nascent RNA synthesis, whereas Mettl3 depletion resulted in transcription upregulation, suggesting that METTL14 might possess an m6A-independent role in gene regulation. We found that METTL14 colocalizes with the repressive H3K27me3 modification. Mechanistically, METTL14, but not METTL3, binds H3K27me3 and recruits KDM6B to induce H3K27me3 demethylation independent of METTL3. Depletion of METTL14 thus led to a global increase in H3K27me3 level along with a global gene suppression. The effects of METTL14 on regulation of H3K27me3 is essential for the transition from self-renewal to differentiation of mESCs. This work reveals a regulatory mechanism on heterochromatin by METTL14 in a manner distinct from METTL3 and independently of m6A, and critically impacts transcriptional regulation, stemness maintenance, and differentiation of mESCs.
Animals ; Mice ; Methylation ; Chromatin ; Histones/metabolism* ; RNA, Messenger/genetics* ; Methyltransferases/metabolism* ; RNA/metabolism*

Objective:This study aimed to investigate the clinical characteristics of patients diagnosed with primary hemophagocytic lymphohistiocytosis (pHLH) associated with perforin gene deficiency.Methods:We retrospectively analyzed the clinical data of 16 pHLH patients associated with perforin gene deficiency at Beijing Friendship Hospital, Capital Medical University, from April 2014 to August 2021. The mutation sites, mutation types, family history, clinical characteristics, and prognosis of the patients were assessed.Results:A total of 16 patients, including ten males and six females, with a median onset age of 17.5 years (range: 4-42 years), were enrolled in this study. Sixteen different mutations were identified, consisting of 11 missense mutations, one nonsense mutation, two frameshift mutations, and two in-frame mutations. All patients harbored at least one deleterious missense mutation, with the most common mutation sites being c.1349C>T (p.T450M) and c.503G>A (p.S168N). Decreased natural killer (NK) cell activity was observed in 11 patients, reduced perforin protein expression in ten patients, concurrent Epstein-Barr virus (EBV) infection at onset in eight patients, a family history in two patients, and central nervous system involvement in four patients. Eleven cases underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT), with eight cases surviving. The median survival time of non-transplanted patients was eight months (range: 4-18 months), while that of transplanted patients was reported as "not reached".Conclusions:Emphasizing the diagnosis of pHLH in adults with perforin gene deficiency. In addition, it should be noted that EBV infection can potentially act as a triggering factor in such disease, and allo-HSCT exerts a substantial effect on the prognosis of patients.

Objective::Microchannels are associated with the progression of atherosclerotic vulnerable plaques. However, in patients with culprit optical coherence tomography (OCT)-defined plaque erosion, the knowledge of microchannels and culprit lesion vulnerability is limited. The aim of this study was to investigate culprit lesion characteristics in patients with ST-segment elevated myocardial infarction (STEMI) caused by plaque erosion with and without microchannels using OCT.Methods::In all, 348 STEMI patients with plaque erosion who underwent OCT of the culprit lesion at the 2 nd Affiliated Hospital of Harbin Medical University (Harbin, China) from August 2014 to December 2017 were included and divided into the microchannel group ( n= 116, 33.3%) and no-microchannel group ( n = 232, 66.7%). The clinical characteristics and OCT-derived plaque features were compared between both groups. Results::Among the 348 STEMI patients with plaque erosion, culprit lesions with microchannels had higher incidence of lipid plaque (59.5% vs. 45.3%, P= 0.012); calcification (41.4% vs. 24.6%, P= 0.002); spotty calcification (30.2% vs. 18.1%, P= 0.014); macrophages accumulation (72.4% vs. 45.7%, P < 0.001); and cholesterol crystals (32.8% vs. 14.2%, P < 0.001) than those without microchannels. In addition, minimal lumen area was smaller ((1.9 ± 0.9) mm 2vs. (2.8 ± 2.3) mm 2, P < 0.001) and lumen area stenosis was greater ((71.3% ± 13.4%) vs. (65.3% ± 19.3%), P= 0.001) in the microchannel group than in the no-microchannel group. Conclusion::In patients with STEMI caused by plaque erosion, one-third manifested typical microchannel characteristics, and those with microchannels were associated with more severe luminal stenosis and more vulnerable plaque features than those without microchannels.

Objective::Microchannels are associated with the progression of atherosclerotic vulnerable plaques. However, in patients with culprit optical coherence tomography (OCT)-defined plaque erosion, the knowledge of microchannels and culprit lesion vulnerability is limited. The aim of this study was to investigate culprit lesion characteristics in patients with ST-segment elevated myocardial infarction (STEMI) caused by plaque erosion with and without microchannels using OCT.Methods::In all, 348 STEMI patients with plaque erosion who underwent OCT of the culprit lesion at the 2 nd Affiliated Hospital of Harbin Medical University (Harbin, China) from August 2014 to December 2017 were included and divided into the microchannel group ( n= 116, 33.3%) and no-microchannel group ( n = 232, 66.7%). The clinical characteristics and OCT-derived plaque features were compared between both groups. Results::Among the 348 STEMI patients with plaque erosion, culprit lesions with microchannels had higher incidence of lipid plaque (59.5% vs. 45.3%, P= 0.012); calcification (41.4% vs. 24.6%, P= 0.002); spotty calcification (30.2% vs. 18.1%, P= 0.014); macrophages accumulation (72.4% vs. 45.7%, P < 0.001); and cholesterol crystals (32.8% vs. 14.2%, P < 0.001) than those without microchannels. In addition, minimal lumen area was smaller ((1.9 ± 0.9) mm 2vs. (2.8 ± 2.3) mm 2, P < 0.001) and lumen area stenosis was greater ((71.3% ± 13.4%) vs. (65.3% ± 19.3%), P= 0.001) in the microchannel group than in the no-microchannel group. Conclusion::In patients with STEMI caused by plaque erosion, one-third manifested typical microchannel characteristics, and those with microchannels were associated with more severe luminal stenosis and more vulnerable plaque features than those without microchannels.