Purpose: Our previous study showed that circulating tumor cell (CTC) count combined with gene mutation detection might help differentiate benign and malignant pulmonary nodules (PNs). Herein, we aimed to expand the study cohort and conduct further sequencing analysis. Materials and Methods: Patients with PNs were included, and CTCs were identified before operation. Low-coverage whole-genome sequencing (LC-WGS) and lung cancer-related targeted gene sequencing were performed on CTCs. The diagnostic efficacy was evaluated by receiver operating characteristic (ROC) curve. The differences in CTC counts among subgroups classified by demographic–clinical characteristics were analyzed. LC-WGS–based copy number variation (CNV) analysis and targeted gene mutation analysis were conducted. Results: A total of 172 patients were included. CTC count of 2.5 was identified by the ROC curves as the optimal diagnostic cutoff. The sensitivity and specificity of CTC count for differentiating benign and malignant PNs were 54.2% and 78.6%, respectively. The diagnostic sensitivity and specificity of combined CTC count, radiological nodule type, and any malignant imaging features were 84.7% and 71.4%, respectively. The CTC counts were significantly greater in patients with aggressive tumors, later stage, and spread through air spaces. CTCs from malignant cases had more CNVs than those from benign cases. Conclusion CTC count can be used in identifying malignant PNs. The diagnostic efficacy can be improved if combined with computed tomography imaging characteristics. Further CNV analysis might help differential diagnosis. Greater CTC count might suggest more aggressive tumors. CTC detection can provide important information and guidance for subsequent management of PNs.

Objective To explore the clinical features of fulminant type 1 diabetes mellitus (FT1DM). Methods The clinical data of 6 patients with FT1DM who were hospitalized in Jinshan Hospital of Fudan University from April 2020 to August 2024 were retrospectively analyzed. Their data were compared with that of 30 patients diagnosed with non-fulminant type 1 diabetes mellitus (NFT1DM) and diabetic ketosis or diabetic ketoacidosis (DKA) who were admitted to the hospital during the same period. The clinical characteristics of FT1DM were summarized. Results All 6 patients with FT1DM were male, with a disease course of 2.00 (1.75, 4.00) d. Three cases exhibited a history of prior infection, four tested positive for glutamic acid decarboxylase antibody (GADA), and five developed severe DKA. The glycated hemoglobin A_1C (HbA_1C) was (6.30±0.67) %, fasting C-peptide (FCP) was 0.07 (0.03, 0.15) ng/mL, 2-hour postprandial C-peptide (2h-CP) was 0.09 (0.03, 0.16) ng/mL. At discharge, all 6 patients received 4-injection insulin regimen, with a dose (0.69±0.15) U·kg⁻¹·d⁻¹. The body mass index (BMI), blood glucose/HbA_1C, blood potassium/HbA_1C, fasting plasma glucose (FPG), 2-hour postprandial plasma glucose (2h-PG), high-sensitivity C-reactive protein (hs-CRP), alanine aminotransferase (ALT), serum creatinine, and blood potassium levels in the FT1DM group were higher than those in the NFT1DM group (P＜0.05), while HbA_1C and glycated albumin (GA) levels were lower than NFT1DM group (P＜0.05). Conclusions FT1DM usually presents with an acute onset of DKA, may be accompanied by a history of preceding infection, and GADA can be positive. Patients with FT1DM have elevated blood glucose/HbA_1C, blood potassium/HbA_1C, FPG, 2h-PG, hs-CRP, ALT, serum creatinine, blood potassium levels, and require insulin therapy, while the HbA_1C and GA levels are lower.

Objective To understand the challenges faced by elderly patients with multiple chronic conditions in community settings during the process of disease management,and to provide a basis for developing intervention strategies for managing comorbidities in the elderly.Methods A phenomenological research approach was adopted,using purposive sampling.From July to October 2024,16 elderly patients with multiple chronic conditions from 5 community service centers in Shenzhen were selected as study participants.Semi-structured in-depth interviews were conducted,and the interview data were analyzed using Colaizzi's seven-step method.Results Totally 4 themes and 8 sub-themes were extracted,including the disorder of multi-source health information structure and environment(insufficient systematic integration of health guidance,uneven quality of knowledge under multiple information channels),the exhaustion of self-management efficacy among chronic disease patients(the coexistence of multiple diseases aggravates the burden of physical and mental symptoms;negative emotions weaken the efficacy of active management),the absence of an integrated management mechanism for multiple diseases(repeated medical visits lead to poor treatment experience;multiple medications pose potential health risks),and the lack of in-depth and dynamic social support practices(formal operation of community support systems,family support facing the dilemma of filial piety and control).Conclusion Elderly patients with multiple chronic conditions in community settings face numerous challenges in disease management.Current chronic disease management in China primarily focuses on single diseases.It is recommended that healthcare professionals shift their perspective,improve the identification,assessment,nursing,and treatment of comorbidities in the elderly,and provide integrated chronic disease management approaches.This would enable a"one-stop"solution to the management challenges of multimorbidity in elderly patients,thereby enhancing their quality of life.

Objective:To investigate the expression characteristics of Zeste enhancer of Zeste homolog 2 (EZH2) in breast cancer tissue and its influence on tumor progression and prognosis.Methods:Transcriptome data of breast cancer tissue and normal breast tissue adjacent to cancer as well as clinical data of patients were obtained from the cancer genome atlas (TCGA) database, gene expression comprehensive database and European genome phenotype archives database, and the difference of EZH2 expression was analyzed using TIMER 2.0 platform. The survival information of breast cancer patients was obtained from the Kaplan Meier Plotter database, and the overall survival time, relapse free survival time and distant metastasis free survival time of breast cancer patients with low EZH2 expression and high EZH2 expression were compared. Select 14 nude mice were selected and randomly divided into si-EZH2 group and control group, with 7 mice in each group.MCF7 culture suspensions transfected with EZH2 knockdown plasmid and control plasmid were inoculated for corresponding group. The body mass and tumor volume of two groups of nude mice inoculated with MCF7 cells were compared at different times. On the 28th day, the nude mice were euthanized and the tumors were dissected to compare the tumor mass of the two groups of nude mice. The normally distributed quantitative data was represented by xˉ ± s. Two independent sample t-tests were used for comparison between two groups, repeated measures ANOVA was used for comparison of body mass and tumor volume between two groups of nude mice at different times, and Bonferroni test was used for pairwise comparison. The comparison of survival rates was conducted using log rank test. Results:A total of 1085 breast cancer tissues and 291 normal adjacent breast tissues were included in the TCGA database. EZH2 expression in breast cancer tissues was higher than that in normal adjacent breast tissues ( P<0.05). In the Kaplan Meier Plotter database, the total survival time, relapse free survival time, and distant metastasis free survival time of breast cancer patients in the EZH2 overexpression group were shorter than those in the EZH2 low expression group ( P=0.013, <0.001, <0.001). After 7 days of inoculation with MCF7 culture suspension, significant subcutaneous tumors were observed on the left back of both groups of nude mice. On the first day, there were no statistically significant difference in body mass between the two groups of nude mice ( P>0.05); On day 7, 13, 19, 25, and 28, the body mass and tumor volume of both groups of nude mice gradually increased (nude mouse body mass: within group F=29.31, P<0.001, between groups F=234.32, P<0.001, Finteraction=16.83, P<0.001; Tumor volume: within group F=34.00, P<0.001, between groups F=193.17, P<0.001, Finteraction=35.61, P<0.001). And the body mass of the siEZH2 group nude mice was higher than that of control group (all P<0.05). On days 19, 25, and 28, tumor the volume of the siEZH2 group nude mice was smaller than that of control group (all P<0.05). On the 28th day, the mass of tumors dissected in the siEZH2 group of nude mice was lower than that in the control group [(0.30±0.07) g vs. (0.61±0.14) g, t=5.16, P<0.001]。 Conclusions:EZH2 is highly expressed in breast cancer tissues and is significantly associated with poor prognosis. Knockdown of EZH2 can significantly inhibit the proliferation and tumor formation of breast cancer cells.

Objective: To accurately determine the ABO blood group of samples exhibiting forward/reverse grouping discrepancies by combining first-generation (Sanger) and third-generation (long-read) sequencing technologies. Methods: Five samples with ABO forward/reverse grouping discrepancies were selected. Serological testing was conducted using automated blood typing instruments and the tube method. Genotyping was conducted using both Sanger and long-read sequencing technologies. Results: Sanger sequencing identified specific genetic mutations in two samples, with genotypes of ABO BA. 04/ABO O.01.01 and ABO B3.05/ABO O.01.02. Further analysis with long-read sequencing revealed specific mutations in the +5.8kb region of intron 1 (c.28+5885C>T and c.28+5861T>G) in three samples where mutations were not detected by Sanger sequencing. These mutations affect the expression of the ABO antigens and are likely responsible for the ABO subgroup phenotypes. Conclusion: The integration of Sanger and long-read sequencing technologies effectively identifies genetic variations causing ABO subtypes, providing a scientific basis for enhancing clinical transfusion safety and ensuring accurate blood group determination.

Objective: To accurately determine the ABO blood group of samples exhibiting forward/reverse grouping discrepancies by combining first-generation (Sanger) and third-generation (long-read) sequencing technologies. Methods: Five samples with ABO forward/reverse grouping discrepancies were selected. Serological testing was conducted using automated blood typing instruments and the tube method. Genotyping was conducted using both Sanger and long-read sequencing technologies. Results: Sanger sequencing identified specific genetic mutations in two samples, with genotypes of ABO BA. 04/ABO O.01.01 and ABO B3.05/ABO O.01.02. Further analysis with long-read sequencing revealed specific mutations in the +5.8kb region of intron 1 (c.28+5885C>T and c.28+5861T>G) in three samples where mutations were not detected by Sanger sequencing. These mutations affect the expression of the ABO antigens and are likely responsible for the ABO subgroup phenotypes. Conclusion: The integration of Sanger and long-read sequencing technologies effectively identifies genetic variations causing ABO subtypes, providing a scientific basis for enhancing clinical transfusion safety and ensuring accurate blood group determination.

In maintaining normal function and activation processes, glycolysis, lipid metabolism, and amino acid metabolism play key roles in the energy demand of platelets. In the resting state, platelets primarily rely on glycolysis and aerobic oxidation to generate energy. Upon activation, platelets preferentially utilize glycolysis, as it can more rapidly provide the required ATP. In addition to glycolysis, platelets can also utilize glycogen and fatty acids as additional energy sources. The ATP provided by fatty acid oxidation is crucial for platelet activation. Additionally, during platelet storage, distinctive changes in energy metabolism occur. In the early stages of storage, platelets primarily rely on glycolysis and the pentose phosphate pathway (PPP) to generate energy. In the mid-storage phase, there is an increase in tricarboxylic acid cycle (TCA) metabolism. In the later stages of storage, cellular metabolism gradually declines. The regulation and flexibility of these metabolic pathways play a critical role in the survival and function of platelets in different states.

[Objective] To compare the characteristics of platelet-rich plasma derived exosomes (PRP-Exos) under different activation conditions and their differential effects on the proliferation capacit of human microvascular endothelial cells (HMEC-1) and human skin fibroblasts (BJ). [Methods] Ten healthy volunteers were recruited, and 10 mL of venous blood anticoagulated with EDTA-K2 was collected. PRP-Exos were prepared and extracted by the secondary centrifugation method. Transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blotting (WB) detection techniques were used to compare the morphological characteristics, particle size distribution, concentration, and the expression of surface marker proteins of PRP-Exos. Through in vitro cell culture, the differences in morphological changes and proliferation abilities of HMEC-1 and BJ cells induced by PRP-Exos in different activator groups were compared. [Results] 1) TEM observation showed that the morphologies of PRP-Exos in different activator groups were different. 2) NTA analysis showed that compared with the particle size of (109.60±2.71) nm in the normal saline group, the particle sizes of the thrombin group [(104.93±1.55) nm] and the mixed agent group [(103.83±1.58) nm] were relatively smaller, while the particle size of the calcium gluconate group [(113.57±4.70) nm] was larger and its particle size distribution range was wider. There were statistically significant differences among different groups (F=7.019, P<0.05). The concentration of exosomes obtained in the group with the mixture of calcium gluconate and thrombin was the highest [(4.87±0.63)×10⁶ particles/mL, P<0.001]. Both the thrombin group [(2.75±0.13)×10⁶ particles/mL, P < 0.01] and the calcium gluconate group [(3.82±0.06)×10⁶ particles/mL, P<0.001] obtained higher concentrations of exosomes compared with the normal saline group. The concentration in the calcium gluconate group was slightly higher than that in the thrombin group, and there was a significant difference between the two groups (P<0.05). 3) WB analysis showed that CD41, CD81, and TSG101 were expressed on the surface of PRP-Exos, and the protein signal intensities of CD41 and TSG101 in the group with the mixture of thrombin and calcium gluconate were the strongest (P<0.001). 4) The results of in vitro cell culture showed that PRP-Exos in the group with the mixture of thrombin and calcium gluconate could significantly promote the proliferation of HMEC-1 and BJ cells (P<0.05). [Conclusion] The PRP treated with the thrombin and calcium gluconate mixture group yielded the highest concentration of exosomes; under in vitro culture conditions, PRP-Exos from this group significantly promoted the proliferation of HMEC-1 and BJ cells.

Extraintestinal manifestations (EIM) of inflammatory bowel disease (IBD) may affect multiple organ systems, with approximately 50% of IBD patients presenting with at least one EIM during the course of their disease, with cutaneous involvement being particularly common. Cutaneous manifestations can present in various forms. This paper reports a case of ulcerative colitis (UC) complicated with bullous pemphigoid (BP), aiming to enhance clinicians' awareness of skin lesions associated with UC.

Objective:To analyze the epidemiological and clinical characteristics of respiratory pathogens in China.Methods:This study was a cross-sectional study, which encompassed 19 core units of the clinical pathogen network and established a three-tiered clinical pathogen surveillance system. Thirty respiratory samples were collected every two weeks from various units from January to December 2024, and the clinical and pathogen diagnostic information were gathered. A total of 11 864 samples were tested using this system. The tier-1 clinical pathogen surveillance system covered influenza A virus (Flu-A), influenza B virus (Flu-B), respiratory syncytial virus (RSV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The tier-2 clinical pathogen surveillance system focused on 18 key respiratory pathogens. The tier-3 clinical pathogen surveillance system further clarified whether any emerging infectious diseases had occurred.Results:The tier-1 clinical pathogen surveillance system showed Flu-A predominated in December, Flu-B predominated in January, SARS-CoV-2 peaked in March and August, whereas RSV circulated sporadically throughout the year. Geographic trends were broadly consistent across the seven major regions, although Flu-A detection in December was notably higher in Northeast China (48.1%(111/231)) and East China (36.2%(148/409)), and RSV detection was concentrated in the Northwest and South China from January to March. Data from the tier-2 clinical pathogen surveillance system indicated that Streptococcus pneumoniae, Mycoplasma pneumoniae, rhinovirus, and adenovirus were detected year-round, of these, Streptococcus pneumoniae and rhinovirus showed elevated positive detection rates from August to September, while adenovirus peaked in January. Legionella pneumophila was not detected throughout the year, and other pathogens fluctuated throughout the year without a consistent pattern. The predominant etiologic agents of pediatric pneumonia were Mycoplasma pneumoniae (35.0%(105/300)), rhinovirus (25.7%(77/300)), and adenovirus (17.3%(52/300)), whereas adult pneumonia was mainly caused by Streptococcus pneumoniae (10.5%(29/277)), Staphylococcus aureus (6.9%(19/277)), Mycoplasma pneumoniae (6.9%(19/277)), and Flu-A (6.1%(17/277)). The tier-3 clinical pathogen surveillance system did not identify any emerging respiratory pathogens. Conclusion:Respiratory pathogens in China in 2024 exhibit distinct temporal and spatial distribution patterns and vary among different populations.