Objective To construct a prognostic model of lung adenocarcinoma(LUAD)based on m5C modification-related genes and to explore its clinical value.Methods Based on the LUAD data in TCGA,GSE30219,GSE31210,and GSE50081 cohorts,prognosis-related m5C modification-related genes were screened,and the prognostic model was constructed by using univariate Cox,Lasso,and multivariate Cox regression analyses.Kaplan-Meier curve,ROC curve,and Cox regression were used to observe the robustness and prognostic performance of the model.The correlation between the prognostic model and clinico-pathologic features was further explored.Results A prognostic model consisting of eight m5C modifi-cation-related genes,including CDK1,CDKN1A,NOP2,RRM2,TCL6,TLR8,TRDMT1,and YTHDF2,was constructed.Risk score was an independent risk factor for the prognosis of patients with LUAD,and it is combined with age,T stage,and N stage to constitute a nomogram which can accurately predict the prognosis of patients.The infiltration of macrophages and CD4+/CD8+T cells was significantly reduced in high-risk patients.The risk score in LUAD tissues was significantly higher than that in normal tissues and was positively correlated with T stage and N stage.The risk score of smoking and EGFR wild-type patients was higher than that of non-smoking and EGFR-mutant patients.Conclusion The prognostic model constructed based on m5C modification-related genes has shown good accuracy and stability in predicting the prognosis of patients with LUAD,and it is closely related to clinical features,driver gene mutations,and immune infiltration,which can provide a potential basis for the treatment and prognostic assessment of LUAD.

Extensive epigenetic reprogramming involves in memory CD8+ T-cell differentiation. The elaborate epigenetic rewiring underlying the heterogeneous functional states of CD8+ T cells remains hidden. Here, we profile single-cell chromatin accessibility and map enhancer-promoter interactomes to characterize the differentiation trajectory of memory CD8+ T cells. We reveal that under distinct epigenetic regulations, the early activated CD8+ T cells divergently originated for short-lived effector and memory precursor effector cells. We also uncover a defined epigenetic rewiring leading to the conversion from effector memory to central memory cells during memory formation. Additionally, we illustrate chromatin regulatory mechanisms underlying long-lasting versus transient transcription regulation during memory differentiation. Finally, we confirm the essential roles of Sox4 and Nrf2 in developing memory precursor effector and effector memory cells, respectively, and validate cell state-specific enhancers in regulating Il7r using CRISPR-Cas9. Our data pave the way for understanding the mechanism underlying epigenetic memory formation in CD8+ T-cell differentiation.
CD8-Positive T-Lymphocytes/metabolism* ; Cell Differentiation ; Chromatin/immunology* ; Animals ; Mice ; Immunologic Memory ; Epigenesis, Genetic ; SOXC Transcription Factors/immunology* ; NF-E2-Related Factor 2/immunology* ; Mice, Inbred C57BL ; Gene Regulatory Networks ; Enhancer Elements, Genetic

Objective:To establish an automatic classification approach for bone marrow cells based on microscopic hyperspectral imaging and three-dimensional spectral convolutional neural network (Spec-CNN).Methods:The research type is establishment of methodology. The study included 306 newly diagnosed patients' bone marrow smears under Wright's staining from the Department of Hematology of the First Medical Center of the PLA General Hospital from November 1st, 2013 to April 30th, 2024. The high-spectrum data and 4k image data of bone marrow cells were simultaneously collected using a microscopic hyperspectral-4k optical path integrated imaging system (with a spectral resolution of 400—1 000 nm). The high-spectrum data was used for model training, while the 4k image data recognized by morphologists was only used as a reference for labeling the high-spectrum data. The high-spectrum data set was divided into training set, validation set and test set in a ratio of 14∶6∶5. The training set and validation set were used to train and fine-tune the Spec-CNN model, and the test set was used to evaluate the model performance. The sensitivity, specificity ,accuracy ,and Kappa coefficient were calculated for comparing the manual annotation results as gold standard with the intelligent identification results of the Spec-CNN model. Five non-data set samples were used for external validation.Results:The acquired hyperspectral data and 4k imaging dataset comprised of 32 categories and 64 800 bone marrow cells. In the test set, the Spec-CNN model demonstrated weighted-average indicators on classification metrics across 32 cell types: sensitivity 87.79%, specificity 99.31%, and accuracy 98.78%, and Kappa coefficient 0.869. For external validation, the mean correct identification rate of bone marrow cells reached 83.28%.Conclusion:We successfully established an automatic classification method of bone marrow cells based on microscopic hyperspectral imaging and three-dimensional Spec-CNN. This method has a good automatic classification ability for 32 types of bone marrow nucleated cells, which has a certain auxiliary effect on improving the diagnosis efficiency of blood diseases for bone marrow morphologists.

Objective To construct a prognostic model of lung adenocarcinoma(LUAD)based on m5C modification-related genes and to explore its clinical value.Methods Based on the LUAD data in TCGA,GSE30219,GSE31210,and GSE50081 cohorts,prognosis-related m5C modification-related genes were screened,and the prognostic model was constructed by using univariate Cox,Lasso,and multivariate Cox regression analyses.Kaplan-Meier curve,ROC curve,and Cox regression were used to observe the robustness and prognostic performance of the model.The correlation between the prognostic model and clinico-pathologic features was further explored.Results A prognostic model consisting of eight m5C modifi-cation-related genes,including CDK1,CDKN1A,NOP2,RRM2,TCL6,TLR8,TRDMT1,and YTHDF2,was constructed.Risk score was an independent risk factor for the prognosis of patients with LUAD,and it is combined with age,T stage,and N stage to constitute a nomogram which can accurately predict the prognosis of patients.The infiltration of macrophages and CD4+/CD8+T cells was significantly reduced in high-risk patients.The risk score in LUAD tissues was significantly higher than that in normal tissues and was positively correlated with T stage and N stage.The risk score of smoking and EGFR wild-type patients was higher than that of non-smoking and EGFR-mutant patients.Conclusion The prognostic model constructed based on m5C modification-related genes has shown good accuracy and stability in predicting the prognosis of patients with LUAD,and it is closely related to clinical features,driver gene mutations,and immune infiltration,which can provide a potential basis for the treatment and prognostic assessment of LUAD.

Objective:To establish an automatic classification approach for bone marrow cells based on microscopic hyperspectral imaging and three-dimensional spectral convolutional neural network (Spec-CNN).Methods:The research type is establishment of methodology. The study included 306 newly diagnosed patients' bone marrow smears under Wright's staining from the Department of Hematology of the First Medical Center of the PLA General Hospital from November 1st, 2013 to April 30th, 2024. The high-spectrum data and 4k image data of bone marrow cells were simultaneously collected using a microscopic hyperspectral-4k optical path integrated imaging system (with a spectral resolution of 400—1 000 nm). The high-spectrum data was used for model training, while the 4k image data recognized by morphologists was only used as a reference for labeling the high-spectrum data. The high-spectrum data set was divided into training set, validation set and test set in a ratio of 14∶6∶5. The training set and validation set were used to train and fine-tune the Spec-CNN model, and the test set was used to evaluate the model performance. The sensitivity, specificity ,accuracy ,and Kappa coefficient were calculated for comparing the manual annotation results as gold standard with the intelligent identification results of the Spec-CNN model. Five non-data set samples were used for external validation.Results:The acquired hyperspectral data and 4k imaging dataset comprised of 32 categories and 64 800 bone marrow cells. In the test set, the Spec-CNN model demonstrated weighted-average indicators on classification metrics across 32 cell types: sensitivity 87.79%, specificity 99.31%, and accuracy 98.78%, and Kappa coefficient 0.869. For external validation, the mean correct identification rate of bone marrow cells reached 83.28%.Conclusion:We successfully established an automatic classification method of bone marrow cells based on microscopic hyperspectral imaging and three-dimensional Spec-CNN. This method has a good automatic classification ability for 32 types of bone marrow nucleated cells, which has a certain auxiliary effect on improving the diagnosis efficiency of blood diseases for bone marrow morphologists.

【Objective】 To investigate the environmental pollution of blood collection and supply institutions by using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and evaluate its application value. 【Methods】 Colonies of air from blood donation sites, skin puncture sites of blood donors, platelet storage boxes, platelet collection equipment, object surfaces of related experimental consumables and cuff surfaces of staff after disinfection were collected, and typical colonies after cultivation were selected for microbial identification by microbial mass spectrometry and then compared with bacteria results detected in blood components from May 2017 to May 2021. 【Results】 Aseptic growth, the number of colonies ≤4.0 CFU/ dish, and the number of colonies > 4.0 CFU/dish accounted for 21.20%, 62.20% and 16.60%, respectively. The qualified rate from high to low was platelet storage box, bacteria settling in the air of blood donation room after disinfection, platelet collection equipment, skin puncture site of blood donors after disinfection, the surface of platelet consumables and the surface of medical staff's overalls. After disinfection, the blood donors' skin puncture sites were compared with other collection sites, and the t values were 2.0371, 1.508, 2.109, 1.961 and 1.778, respectively, with no significant difference (P>0.05). Thirty cases of bacterial contamination of blood components were detected from May 2017 to May 2021, among which the detection rate of apheresis platelets was the highest, and the t values were 1.731 and 2.272, relative to the contamination frequency of erythrocytes and plasma bacteria (P>0.05), while the t value was 2.875, relative to concentrated platelets, with significant difference (P<0.05). 【Conclusion】 Bacterial contamination of blood components mostly come from air bacteria settling, blood donors' arms and skin after disinfection, and surfaces of related equipment and materials. Therefore, it is of clinical significance to conduct strict disinfection of working sites, establish disinfection monitoring methods and formulate disinfection hygiene standards in blood stations.

IgG4-related disease (IgG4-RD) is a recently recognized disorder characterized by elevated serum IgG4 levels and infiltration of IgG4 positive blood cells in the affected organs. However, other conditions like malignancy as well as connective tissue diseases, may show similar findings. A 56-year-old male patient visited Second Xiangya Hospital, Central South University for recurrent fever and chest pain for more than 1 month. Preliminary tests diagnosed as IgG4-related lung disease (IgG4-RLD). However, the improvement of symptoms was absent after the treatment with methylprednisolone. The patient underwent the second biopsy and the result eventually demonstrated lung adenocarcinoma. The role of IgG4 in the pathogenesis or prognosis for lung adenocarcinoma remains unclear. Therefore, a thorough evaluation of symptoms, test of specific serum markers and eventually pathological confirmation are required to avoid misdiagnosis.

【Objective】 To observe the effect of hydrogen peroxide atomization sterilizer using low concentration hydrogen peroxide disinfectant on the environment and object surface of physical examination area (hereinafter referred to as " physical examination area" ) in blood centers, so as to provide a simple method which is safe, efficient, easy to operate, harmless to human body and has no corrosive effect on equipment. 【Methods】 The physical examination area was disinfected with atomized hydrogen peroxide sterilizer, and the difference of colony number between air and surface before and after disinfection was compared to evaluate the disinfection effect. 【Results】 After disinfection, the hydrogen peroxide residue was detected for 25 times at 5 points, and the results were (0.7~1)ppm, with no statistical difference (P>0.05). 25 tests were carried out at 5 points, and the quartile of the test results was (0~2)CFU/ dish, and the qualified rate was 100%. The test results of bacteria before and after disinfection were statistically significant (P<0.05), which met the requirements of Class Ⅱ environment in Hygienic Standard for Hospital Disinfection(GB15982-2012). After disinfection, the quartile of surface colony detection results of workbench, blood donor seat, screen and door handle were (0~24.1)CFU/cm^2, (1.6~55.4)CFU/cm^2, (0~7.2)CFU/cm² and (0~4.8)CFU/cm^2, with the qualified rate at 80%, 48%, 100% and 100%, respectively, which were in accordance with the requirements of Class Ⅲenvironment in GB15982-2012. The number of colonies after disinfection at the above detection sites decreased significantly compared with that before disinfection (P<0.05). The surface contact plate pressing method and cotton swab smearing method were used to detect the number of colonies on the surface of sterilized work tables and blood donor seats, and the detection rate of the former was higher than that of the latter, with statistical significance (P<0.05). 【Conclusion】 After disinfection by hydrogen peroxide atomization sterilizer, the hydrogen peroxide residue met the requirements specified in the manual. The terminal disinfection effect of air in the physical examination area environment can meet the Class Ⅱ environmental requirements of GB15982-2012. However, the number of microorganisms on object surface after terminal disinfection was significantly lower than that before disinfection.

Primary enteric adenocarcinoma is a rare variant of primary pulmonary adenocarcinoma. This disease lacks a distinctive manifestation and often requires pathological examination to make a definite diagnosis. A male patient visited the Second Xiangya Hospital, Central South University for consistent cough and sputum production for about 1 year. Anti-infection therapy was given but it showed ineffectiveness. Enteric adenocarcinoma was diagnosed after percutaneous lung biopsy according to pathological findings. Combining this case with relevant literature, we summarized the characteristics to raise physicians' awareness for this rare subtype.
Adenocarcinoma/surgery* ; Adenocarcinoma of Lung ; Humans ; Lung ; Lung Neoplasms ; Male

Objective: To investigate the effects of 1, 25-dihydroxy vitamin D₃[1, 25(OH)₂D₃] on food allergy(FA) in mice and its mechanism. Methods: A total of 40 BALB/c mice were randomly divided into 5 groups, 8 in each group, including control group (group C) and FA model group (FA group), according to the dose of 1, 25(OH)₂D₃ intervention, the mice of the FA group were divided into FA₀ group (0), FA_l group [10 μg/(kg·d)], FA_m group [50 μg/(kg·d)] and FA_h group[100 μg/(kg·d)]. Egg albumin was used to establish a food allergy model, with different doses of 1, 25(OH)₂D₃ for gastric intervention, and the control group was replaced by 9 g/L saline.The serum levels of ovalbumin-immunoglobulin E(OVA-IgE), interleukin(IL)-9 and IL-17 of mice were measured by using enzyme linked immunosorbent assay after the last excitation, and HE staining and histopathological examination were carried out in the small intestine of mice. Results: Compared with group C, FA₀ group and FA_h group small intestinal mucosa in mice had different degrees of damage, partial peeling off, structure disorder, villi epithelial cell focal falls peeling off, necrosis, lamina propria edema, congestion, a large number of inflammatory cells infiltration, low but the FA_l group and FA_m group had light mucosa damage, intestinal epithelial basically intact, with integrity, no congestion, edema, and inflammatory cells infiltration to a lesser degree.The mean concentrations of serum IgE, IL-9 and IL-17 in different groups were statistically significant (F=40.770, 9.530, 5.624, all P<0.05). Compared with the FA₀ group [(41.87±3.19) ng/L], the OVA-IgE of the FA_l group [(22.71±4.77) ng/L] and the FA_m group [(16.34±2.81) ng/L] were significantly reduced (t=5.533, 11.835, all P<0.01), but there was no significant difference between the FA_h group [(36.29±6.52) ng/L] (t=1.673, P>0.05). Compared with the FA₀ group [(161.77±50.44) ng/L], the IL-9 levels of the FA_l group [(94.29±18.79) ng/L] and the FA_m group[(84.45±30.88) ng/L] were significantly lower (t=3.267, 3.366, all P<0.01), while that of the FA_h group [(36.29±6.52) ng/L] was not significantly lower (t=0.777, P>0.05). Compared with FA₀ group [(81.55±29.37) ng/L], IL-17 levels of FA_h group [(133.58±47.05) ng/L] was significantly increased (t=2.653, P<0.05), while IL-17 level of FA_l group [(79.41±25.15) ng/L] and FA_m group [(58.81±26.00) ng/L] were lower than that of FA₀ group [(81.55±29.37) ng/L], but the difference was not statistically significant (t=0.154, 1.640, all P>0.05). Conclusions Low, medium dose of 1, 25(OH)₂D₃ supplements can inhibit mice food allergies, but high doses of 1, 25(OH)₂D₃ improve performance in mice food allergies, and 1, 25(OH)₂D₃′s influence on the secretion of IL-9 is one that influences mechanism of food allergy.