Objective:To establish a flow cytometric assay for myeloid-derived suppressor cells (MDSC) in human peripheral whole blood and reference intervals for healthy adults in Shanghai region.Method:A whole blood eight-color and twelve-parameter flow cytometric assay was designed, utilizing fluorescently labeled antibodies against CD45, CD3, CD19, CD123, CD56, CD16, HLA-DR, CD33, CD11b, CD14, CD15 and CD20.A total of 246 healthy participants who met the health standards from the health check-ups conducted at the Tongji University Affiliated Shanghai East Hospital between May 8 to December 2, 2024 were enrolled. Peripheral venous whole blood was collected using EDTA-K 2 anticoagulant vacuum tubes for MDSC detection. A single-platform flow cytometry based relative count technique was used to quantify the percentage of each MDSC subpopulation. Kolmogorov Smirnov (K-S) test was used to test the distribution of specimens. Mann-Whitney U test and Kruskal-Wallis (K-W) test were used to evaluate whether reference intervals should be established separately based on gender or age. According to the clinical significance of MDSC, bilateral reference intervals were taken. Non parametric methods were used to take the percentile P2.5 and P97.5 to represent the rank of the lower and upper reference limits, respectively. Results:The results showed that a gating strategy was designed to exclude granulocytes, lymphocyte lineage cells, and natural killer cells. The K-S test results showed that the MDSC in each group of healthy individuals were distributed in a skewed manner. The U test showed significant gender differences ( P0.05) in the distribution of total myeloid-derived suppressor cells (T-MDSC) and polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC). The K-W test showed no significant differences in MDSC among different age groups (21-30 years old, 30-40 years old, 40-50 years old, and 50-60 years old). T-MDSC reference interval is 0.056%-0.485%, PMN-MDSC reference interval is 0.035%-0.406%, Monocytic myeloid-derived suppressor cells (M-MDSC) reference interval is 0.000%-0.221%, early-stage myeloid-derived suppressor cells (E-MDSC) reference interval is 0.004%-0.125%. Reference interval verification was conducted on 20 healthy individuals, with a pass rate of 100%. Conclusion:A whole blood eight-color and twelve-parameter flow cytometric assay was established in this experiment. Based on the flow cytometry single platform method, reference intervals for healthy adults in Shanghai region were established.

Objective:To explore the clinical phenotypes and hematological characteristics of β-thalassemia combined with α-globin gene triplet.Methods:A retrospective case analysis study was conducted, taking individuals diagnosed with thalassemia who sought for outpatient services in No 1 people′s hospital of Chenzhou affiliated to South China University from August 10, 2021, to December 31, 2023 as study objectives. Among them, there were 8 768 males and 11 707 females, aged 31.5 (23.0, 46.0) years old. Blood analysis were analyzed by hematology analyze.The hemoglobin(Hb) Hb A, HbA 2,Hb F bands were analyzed by Capillary electrophoresis method, and the genotypes of thalassemia were analyzed by high-throughput sequencing technology.Hematological parameters between different genotypes of thalassemia were analyzed using t-tests and calibrated t-tests for data analysis. Results:A total of 27 cases of beta thalassemia combined with alpha globin triplet were detected, The average hemoglobin (Hb) of 11 cases of β Codon41/42 (-CTTT)/β N combined with ααα anti 4.2 (3.7) (92±16)g/L was lower than that of β Codon41/42 (-CTTT)/β N(112±11)g/L, and the difference was statistically significant ( t=3.97, P0.05). The average Hb of 8 cases of β IVS-Ⅱ-654 (CT)/β N combined with ααα anti 4.2 (3.7)(85±21) g/L was lower than that of β IVS-Ⅱ-654 (CT)/β N(116±12) g/L, and the difference was statistically significant ( t=4.05, P0.05). Conclusion:When the mutation site is at Codon41/42 (-CTTT) or IVS-Ⅱ-654 (CT), β-thalassemia combined with alpha globin triplet can make the clinical manifestations of β-thalassemia at this site more pronounced.

A 71-year-old male patient was admitted to the Hematology Department of the Second Hospital of Hebei Medical University due to abdominal pain accompanied of chest tightness and shortness of breath, which appeared without obvious cause over a month ago, and multiple abnormal enlarged lymph nodes around the stomach.After admission, physical examination revealed an anemic appearance, serum immunofixation electrophoresis showed positivity for immunoglobulin A (IgA) and κ light chain. The cytological morphology of bone marrow, bone marrow biopsy and lymph node flow cytometry all indicated a significant proliferation of plasma cells.The preliminary clinical diagnosis is multiple myeloma.The review of bone marrow cytology revealed a small number of abnormal cells, which provided direction for the next clinical diagnosis and treatment of lymph node pathological biopsy, immunohistochemical staining, and genetic testing were performed.The patient was ultimately diagnosed with angioimmunoblastic T-cell lymphoma (AITL) with monoclonal gammopathy.After the clinical administration of the chemotherapy regimen consisting of Chidamide and CHOPE.The patient′s symptoms have improved compared with before.The treatment was effective.The AITL is a subtype of nodal T-follicular helper cell lymphoma. Which is a rare type of non-Hodgkin′s lymphoma.The cytological morphology of bone marrow, lymph node pathological biopsy and immunohistochemistry provided important basis and ideas for the correct diagnosis and treatment of the patient.

A case of 75-year-old male patient was admitted to the Second Affiliated Hospital of Harbin Medical University on June ,2024 complaining 'repeated cough for half a year', accompanied by night fever, night sweats, fatigue, weight loss. He was initially diagnosed as diffuse large B-cell lymphoma through chest CT and bronchoscopic biopsy, then confirmed as diffuse large B-cell lymphoma with MYD88 genetic aberration with bone marrow inconsistent involvement by PET-CT, bone marrow biopsy, immunohistochemistry(IHC),FISH and other tests. The R-CHOP was given as first-line regimen treatment immediately, but the patient appeared allergyic to Rituximab and Zuberitamab sequencially, therefore we change the plan to CDOP for continued chemotherapy and discharged after remission. In his regular admission, we give G-CHOP regimen for chemotherapy, no adverse reactions were found.

Colorectal cancer is a malignant tumor of the digestive system that seriously endangers human health. Small extracellular vesicles (sEV) are a class of extracellular vesicles with lipid bilayer structure with diameter less than 200 nm. SsEV mediate cell-to-cell information exchange through a variety of components. MicroRNA (miRNA), long non-coding RNA (lncRNA), circular RNA (circRNA) and proteins are involved in regulating the occurrence and development of colorectal cancer and play important roles in the diagnosis and prognosis of colorectal cancer. The various active components of sEV hold application value in the 'diagnosis-treatment-prognosis' of colorectal cancer, providing a reference for their application prospects.

Various methods for cancer biomarker detection have been developed to realize cancer early diagnosis, precise treatment, and prognosis evaluation. Traditional techniques such as ELISA (enzyme-linked immunosorbent assay)have been widely applied worldwide, but face challenges in standardized wide application due to molecular-level limitations, sensitivity, and application scope. In 2014, the ultra-sensitive Simoa (Single Molecule Array) technology was introduced, breaking through the shortcomings of small-molecule detection. Simoa enables highly efficient detection and quantitative analysis of biomarkers across multiple fields, providing new insights into cancer biomarker tests. Simoa demonstrates significant advantages in biomarker detection and clinical application, with a wide clinical application prospect in the diagnosis and treatment of lung cancer, breast cancer and other cancers, and provides a theoretical basis for the construction of cancer prediction model.

The blood and circulatory system, which are crucial for maintaining normal physiological functions and homeostasis in the human body, can be subjected to erroneous attacks by the immune system due to autoimmune abnormalities. This can lead to symptoms, including anemia, purpura, myocarditis and arrhythmia, which can severely affect the health and life-quality of patients. Laboratory-tests play a vital role in the diagnosis and treatment of autoimmune diseases, with test items including a variety of biomarkers such as autoantibodies, special proteins, and immune cells. This article reviews the biomarkers related to autoimmune diseases that primarily affect the blood and circulatory system. It also aims to draw the attention of medical professionals to these biomarkers and their associated laboratory tests, and to promote their application in clinical practice.

Objective:To identify strain-specific features of Klebsiella pneumoniae by analyzing metagenomics next generation sequencing (mNGS) data, thereby expanding the downstream applications of mNGS. Methods:The sequences of K.pneumoniae strains were organized from both the self-built database of the long-term multi-center research cohort in China established by the Peking University People′s Hospital from 2009 to 2020 (with 2 345 sequences) and the public databases (with 19 648 sequences). The existing large-scale databases were compressed, and a set of strains representative of clonal groups were screened. A strain genome information library was constructed based on k-mer features, and the most matching representative sequences in the database were searched for the raw mNGS data. The search results of the self-built library and public library were merged and optimized to update the prediction of antimicrobial-resistance characteristics and avoid the impact of uneven data distribution on the results. A total of 314 clinical samples from patients with K.pneumoniae detected by mNGS in the Clinical Microbiology Laboratory of Peking University People′s Hospital from 2022 to 2024 were retrospectively collected, and 101 samples with positive clinical culture results were selected to validate the prediction results. The antimicrobial-resistance phenotypes were verified by clinical antimicrobial susceptibility test results. Whole-genome sequencing was performed on the culture strains of 14 samples randomly selected using random numbers to verify the genotypes. Single nucleotide polymorphism distance analysis was used to verify the occurrence of outbreak events. The χ2 test and Mann-Whitney U test were used for statistical analysis. Results:A representative strain sequence k-mer feature library containing self-built and public sub-libraries was constructed. The library construction required only about 1 hour with <3 GB storage, with a high compression ratio and low update cost. Using k-mer-based analysis, mNGS data achieved precise strain characterization within 4 minutes and and <5 GB memory occupation. There was a significant difference in the antimicrobial-resistance rates to more than half of the antibiotics between the self-built database (90.8%, 2 130/2 345) and the public database (22.7%, 4 457/19 648) ( χ2=4 634.1, P<0.001). After optimizing the search results, the mean category agreement, sensitivity, and specificity of the prediction for eight antibiotics reached 84.8% (323/381), 78.9% (131/166), and 91.2% (196/215), respectively. The target genotypes were successfully detected in 10 out of 12 samples, and two outbreak events (2 samples per event) were successfully identified. Conclusions:An independent analysis process adapted to the needs of identifying the features of K. pneumoniae strains in mNGS data was developed. This process requires minimal computational resources and processing time and can directly achieve the simultaneous analysis of the antimicrobial-resistance phenotypes of K. pneumoniae at the strain level and their corresponding genomic characteristic profiles based on the raw mNGS reads.

Objective:To retrospectively analyze the changes in the proportion of refined lymphocyte subsets in peripheral blood of patients with Graves disease (GD), and their correlation with the clinical characteristics and efficacy of GD, and to explore the immunological pathogenesis of Graves disease for seeking new therapeutic targets.Methods:A total of 97 newly diagnosed GD patients (GD group), 27 patients after treatment (treatment group), and 31 healthy individuals (control group) who visited the Fifth Medical Center of the PLA General Hospital from 2018 to 2021 were included in this study. The data of refined lymphocyte subsets, thyroid function, blood routine and clinical treatment of the three groups were compared and analyzed. The t-test and rank sum test were used to compare the proportions of lymphocyte subsets among different groups, and Pearson correlation analysis was used to analyze the correlation between the proportions of lymphocyte subsets and thyroid function indicators.Results:The proportion of B cells in GD group was higher than that in the control group [16.2%(11.8%, 21.8%) vs 10.2%(8.1%,13.6%)], while the proportion of natural killer (NK) cells was lower [9.4%(4.9%, 13.6%) vs 14.6%(12.1%,18.8%)], and the differences were statistically significant ( P<0.05). Abnormal T cell differentiation: the proportions of functional cells, including activated T cells, memory T cells, clustering antigen(CD)4+memory T cells, Th1 cells, and Tc1 cells, were lower than that in the control group [3.2%(2.1%, 5.7%) vs 5.8%(3.0%, 9.3%), P<0.05; 36.7% (29.9%, 48.1%) vs 48.0%(39.2%,57.7%), P<0.05; 23.1%(17.4%, 30.1%) vs 28.9%(23.3%,34.6%), P<0.05; 16.4% (11.8%, 23.6%) vs 24.3%(16.9%,28.5%), P<0.05; 28.5% (14.7%, 39.2%) vs 46.3%(21.6%,69.2%), P<0.05]. The proportion of activated T cells in the treatment group was higher than that in the GD group [6.5% (4.6%, 13.6%) vs 3.2% (2.1%, 5.7%), P<0.05]. The total triiodothyronine results showed positive correlations with B cells ( r=0.356, P<0.01) and negative correlations with NK cells ( r=?0.416, P<0.01), while the total thyroxine values showed negative correlations with NK cells and activated T cells ( r=?0.318,?0.335; P<0.01). Thyroid stimulating hormone and CD8+initial T cells were positively correlated ( r=0.382, P<0.01). The proportion of B cells, cytotoxic T cells and suppressor T cells in CD8+cells of patients with complications [such as Graves orbitopathy (GO), thyroid toxic cardiomyopathy, etc.] was significantly different from that of the simple GD patients [18.3% (14.1%, 27.1%) vs 14.6% (10.8%, 21.4%), Z=2.54, P<0.05; 73.4%(65.6%,83.6%)vs 65.0%(50.3%,79.3%), Z=2.93, P<0.05; 26.6%(16.4%, 37.5%)vs 35.0%(20.7%,49.7%), Z=?2.74, P<0.05]. The proportion of suppressor T cells in GO patients was lower than that in non-GO patients [6.1% (3.4%, 8.1%) vs 8.5% (4.9%, 13.6%), Z=?3.20 P<0.05]. Conclusion:There are significant alterations in the circulating immune cells of GD patients, suggesting that immunological abnormalities play a crucial role in the onset and progression of the disease.

Objective:To explore the clinical predicting value of serum intestinal fatty acid-binding protein (I-FABP) in the development of esophageal varices (EV) in patients with chronic hepatitis B cirrhosis.Method:We used case-control study. A retrospective analysis was performed on the clinical data of 169 patients with hepatitis B cirrhosis who were admitted to the Affiliated Hospital of Yanbian University from September 2020 to October 2023. EV diagnosis and grades were based on gastroscopy. Enzyme-linked immunosorbent assay was used to measure the serum level of I-FABP on admission. Spearman correlation analysis was used to investigate the correlation among variables. Contributing factors of EV were evaluated using univariate and multivariate logistic regression analysis. Receiver operating characteristic (ROC) curves were used to evaluate the predictive performance of I-FABP for EV presence.Results:The gastroscopy showed 59 patients without EV. The median of I-FABP in the EV Group was significantly higher than that in the no-EV Group [2.01 (1.39, 2.89) μg/L vs 0.96 (0.77, 1.91) μg/L], and the difference was statistically significant ( Z=5.585, P<0.001). I-FABP showed significant positive correlations between model for end-stage liver disease sore and Von Willebrand Factor Antigen/thrombocyte Ratio ( r=0.523, 0.328, both P<0.001). Multiple logistic regression analysis identified I-FABP as the independent factor contributing to the presence of EV ( OR=1.73, P=0.045). The area under the curve of I-FABP predicting EV was 0.76. The cut-off was 1.46 μg/L. Conclusion:I-FABP is a potential marker for the formation of EV in patients with hepatitis B cirrhosis, and its increased concentration is related to reduced hepatic reserve and portal hypertension.