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Background: Immune checkpoint blockade (ICB) therapy, including antibodies targeting the programmed cell death protein 1 (PD-1) pathway, has significantly prolonged the overall survival (OS) in patients with advanced cervical cancer (CC). ICB treatment affects both target cells and various components released by immune cells, which can be observed in peripheral blood. However, there has been limited research on the dynamics of peripheral blood immunophenotyping and its association with OS in CC patients receiving ICB therapy. Methods: Patients with persistent, recurrent, or metastatic CC treated with ICB were enrolled between December 2019 and September 2022. The dynamic changes in peripheral blood immune cells, immunoglobulins, and complement components were analyzed at baseline (within 30 days prior to the first ICB cycle) and after the second cycle of ICB treatment (4–6 weeks after the first ICB treatment). Associations of the baseline levels of peripheral blood immune cells, immunoglobulins, complement components with OS were analyzed using multivariable Cox regression analysis. Results: In this retrospective cohort study, 119 patients who received at least two cycles of ICB were included. Data on peripheral blood immune cells, immunoglobulins, and complement components were available for 70 of these patients. The percentages of suppressor T (Ts) cells and natural killer (NK) cells in peripheral blood increased significantly post-ICB treatment, whereas the Th/Ts ratio and IgM levels decreased. The percentages of cytotoxic T (Tc) cells, Ts cells, the Th/Ts ratio, and levels of IgM, IgA, C3, and C4 were significantly associated with the OS of patients. Furthermore, multivariable Cox regression analysis found that a high level of IgA was associated with poor OS of the patients (HR = 2.918; 95% CI, 1.081–7.877, P = 0.035). Conclusion: Our study demonstrated the potential proliferation of peripheral blood anti-tumor T cells in some CC patients undergoing ICB therapy. The observed associations between peripheral blood immunophenotyping and OS suggest that these biomarkers might have potential as prognostic tools. [ABSTRACT FROM AUTHOR]

Pelvic organ prolapse (POP) is a common gynecological disease caused by pathological defects, lesions, or mechanical weakening of the support structures of the pelvic floor. However, its pathogenesis is unclear. Circular RNAs (circRNAs) are covalently closed, endogenous biomolecules, which are thought to play an important role on skeletal muscle development by regulating gene expression. In this study, five pairs of peripheral blood samples from control and POP groups were used for circRNA sequencing analysis to obtain differential expression profiles. A total of 75 differentially expressed circRNAs (DEcircRNAs) were identified (fold change >2.0, P < 0.05). Furthermore, RT-qPCR confirmed that the expression levels of two circRNAs (hsa_circ_0067962 and hsa_circ_0057051) were significantly lower in the POP group. The two validated DEcircRNAs were abundantly involved in the collagen catabolic process. The circRNA-miRNA-mRNA network of two DEcircRNAs comprised nine mRNAs, which indicated that hsa_circ_0067962 and hsa_circ_0057051 may be involved in the pathogenesis of POP by regulating these nine mRNAs. [ABSTRACT FROM AUTHOR]

Background: Pancreatic cancer remains one of the deadliest malignancies, largely due to its late diagnosis and lack of effective therapeutic targets. Materials and methods: Using traditional machine learning methods, including random-effects meta-analysis and forward-search optimization, we developed a robust signature validated across 14 publicly available datasets, achieving a summary AUC of 0.99 in training datasets and 0.89 in external validation datasets. To further validate its clinical relevance, we analyzed 55 peripheral blood samples from pancreatic cancer patients and healthy controls using qPCR. Results: This study identifies and validates a novel five-gene transcriptomic signature (LAMC2, TSPAN1, MYO1E, MYOF, and SULF1) as both diagnostic biomarkers and potential drug targets for pancreatic cancer. The differential expression of these genes was confirmed, demonstrating their utility in distinguishing cancer from normal conditions with an AUC of 0.83. These findings establish the five-gene signature as a promising tool for both early, non-invasive diagnostics and the identification of actionable drug targets. Conclusion: A five-gene signature is established robustly and has utility in diagnostics and therapeutic targeting. These findings lay a foundation for developing diagnostic tests and targeted therapies, potentially offering a pathway toward improved outcomes in pancreatic cancer management. [ABSTRACT FROM AUTHOR]

Background: Steroid-induced osteonecrosis of the femoral head (SIONFH) is a universal hip articular disease and is very hard to perceive at an early stage. The understanding of the pathogenesis of SIONFH is still limited, and the identification of efficient diagnostic biomarkers is insufficient. This research aims to recognize and validate the latent exosome-related molecular signature in SIONFH diagnosis by employing bioinformatics to investigate exosome-related mechanisms in SIONFH. Method: The GSE123568 and GSE74089 datasets were employed to conduct differentially expressed genes (DEGs) analysis, and the GSE123568 dataset was subjected to perform weighted genes co-expression network analysis (WGCNA). The exosome-related genes (ERGs) were retrieved from the GeneCards database. We identified differentially expressed exosome-related genes (DEERGs) between healthy controls (HC) and SIONFH patients, and a consensus clustering analysis was then implemented to group the SIONFH patients. The CIBERSORT was implemented to calculate the immune cell infiltration. Gene Set Variation Analysis (GSVA), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) were conducted to investigate latent enriched pathways. In addition, machine-learning algorithms were applied to refine the DEERGs. Ultimately, we verified the diagnostic significance and expression of the hub genes using the SIONFH datasets and performing quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis. Results: This study identified twenty DEERGs from the peripheral serum and hip articular cartilage samples of SIONFH patients and HC. Two SIONFH subtypes related to ERGs were identified, and distinctions in pathways and immune cell infiltration patterns were compared. SIONFH's high-risk subpopulation exhibited enriched immune-related pathways and high immune cell infiltration, such as M0 macrophages, resting mast cells, and neutrophils. Three machine-learning algorithms then determined LCP1, PNP, UBE2V1, and ZFP36 as four exosome-related hub genes (ERHGs). Compared to HC samples, these ERHGs showed excellent diagnostic efficiency (overall AUC for ERHGs is in the range of 0.923 to 0.970 in GSE123568) in SIONFH samples. LCP1, PNP, UBE2V1, and ZFP36 expressions were validated in the GSE123568 and GSE74089 datasets and finally detected in peripheral serum samples with accordant expression by RT-qPCR. Conclusion: Twenty potential exosome-related genes involved in SIONFH were identified through bioinformatics analysis. LCP1, PNP, UBE2V1, and ZFP36 might become candidate biomarkers and therapeutic targets because they have an intimate relationship with exosomes. These findings shed light on the exosome-related acquaintance of SIONFH and might contribute to the diagnosis and prognosis of SIONFH. [ABSTRACT FROM AUTHOR]

To ensure the preservation of functional hematopoietic stem cells (HSC) and committed progenitor cells (HPC) at + 4 °C in ex vivo expanded cord blood cell products during worldwide transportation and subsequent infusion—without the need for washing and cell concentration—we developed a conservation medium called Stabilizer of Expanded Cells (SEC), composed exclusively of injectable pharmacological products. The in vivo engraftment assay in immunodeficient mice was used to detect primitive HSCs before and after preservation at + 4 °C. In some experiments, a complex phenotype based on CD34, CD38, and CD133 expression was utilized for this purpose. Committed progenitors (CFU-GM, BFU-E, and CFU-Mix) were detected using methylcellulose culture colony-forming assays. Additionally, in some cases, the energetic metabolism (mitochondrial respiration) was evaluated using Seahorse technology. SEC was able to preserve the functionality of HSCs and HPCs in ex vivo expanded cell populations at + 4 °C for at least 48 h. Furthermore, SEC is also effective in fully preserving HSCs and HPCs in cytapheresis products for at least 72 h. Additionally, SEC enabled the full preservation of HSCs and HPCs for 72 h in freshly collected cord blood, maintaining a normal metabolic profile of CD34+ cells. The SEC medium exhibits a positive effect on the maintenance of both HSCs and HPCs at + 4 °C, regardless of their source. Therefore, SEC can be applied in cell therapy protocols based on HSCs and HPCs with a significant advantage: the product does not need to be washed and concentrated before injection into the patient. [ABSTRACT FROM AUTHOR]

Abstract Background Immune checkpoint blockade (ICB) therapy, including antibodies targeting the programmed cell death protein 1 (PD-1) pathway, has significantly prolonged the overall survival (OS) in patients with advanced cervical cancer (CC). ICB treatment affects both target cells and various components released by immune cells, which can be observed in peripheral blood. However, there has been limited research on the dynamics of peripheral blood immunophenotyping and its association with OS in CC patients receiving ICB therapy. Methods Patients with persistent, recurrent, or metastatic CC treated with ICB were enrolled between December 2019 and September 2022. The dynamic changes in peripheral blood immune cells, immunoglobulins, and complement components were analyzed at baseline (within 30 days prior to the first ICB cycle) and after the second cycle of ICB treatment (4–6 weeks after the first ICB treatment). Associations of the baseline levels of peripheral blood immune cells, immunoglobulins, complement components with OS were analyzed using multivariable Cox regression analysis. Results In this retrospective cohort study, 119 patients who received at least two cycles of ICB were included. Data on peripheral blood immune cells, immunoglobulins, and complement components were available for 70 of these patients. The percentages of suppressor T (Ts) cells and natural killer (NK) cells in peripheral blood increased significantly post-ICB treatment, whereas the Th/Ts ratio and IgM levels decreased. The percentages of cytotoxic T (Tc) cells, Ts cells, the Th/Ts ratio, and levels of IgM, IgA, C3, and C4 were significantly associated with the OS of patients. Furthermore, multivariable Cox regression analysis found that a high level of IgA was associated with poor OS of the patients (HR = 2.918; 95% CI, 1.081–7.877, P = 0.035). Conclusion Our study demonstrated the potential proliferation of peripheral blood anti-tumor T cells in some CC patients undergoing ICB therapy. The observed associations between peripheral blood immunophenotyping and OS suggest that these biomarkers might have potential as prognostic tools.

Yan-Hua Liu,1,&ast; Jin-Wen Su,2,&ast; Jing Jiang,3 Bing-Fen Yang,1 Zhi-Hong Cao,1 Fei Zhai,1 Wen-Na Sun,1 Ling-Xia Zhang,1 Xiao-Xing Cheng1 1Beijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, the Eighth Medical Center of PLA General Hospital, Beijing, 100091, People’s Republic of China; 2Division of Critical Care Medicine, Senior Department of Tuberculosis, the Eighth Medical Center of PLA General Hospital, Beijing, 100091, People’s Republic of China; 3Institute of Research, Beijing Key Laboratory of Organ Transplantation and Immune Regulation, Senior Department of Respiratory and Critical Care Medicine, the Eighth Medical Center of PLA General Hospital, Beijing, 100091, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xiao-Xing Cheng; Ling-Xia Zhang, Beijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment, Institute of Tuberculosis Research, Senior Department of Tuberculosis, Eighth Medical Center of PLA General Hospital, 17 hei Shan Hu Road, Haidian, Beijing, 100091, People’s Republic of China, Fax +86 10 51520496 ; +86 10 66775969, Email xcheng79@outlook.com; zhanglingxia7@163.comPurpose: Tuberculosis (TB) is a major global health threat and its diagnosis remains challenging. This study aimed to develop a nomogram that incorporated peripheral blood transcriptional signatures and other blood tests for the diagnosis of tuberculosis.Patients and Methods: Patients with TB, patients with other definite pulmonary diseases (OPD), individuals with latent tuberculosis infection (LTBI), and healthy controls (HC) were retrospectively enrolled between May 2017 and April 2018. The results of the interferon-γ release assay (IGRA) and blood counts were obtained from medical records, and the transcripts of 10 genes were detected using reverse transcription polymerase chain reaction (RT-PCR). Variable selection was performed using least absolute shrinkage and selection operator regression (LASSO) and multivariate logistic regression was performed for the optimal prediction model with backward direction. The model was displayed as a nomogram, and its performance was evaluated for discrimination ability, calibration ability, and clinical usefulness. Internal validation of the prediction model was conducted using bootstrap resampling.Results: A total of 185 participants were enrolled, including 84 patients with TB and 101 controls. A prediction nomogram composed of IGRA, percentage of neutrophils, and expression levels of CD64, granzyme A (GZMA), and PR/SET domain 1 (PRDM1) was established. The nomogram demonstrated good discrimination, with an unadjusted area under the curve (AUC) of 0.914 (95% CI: 0.875– 0.954) and a bootstrap-corrected AUC of 0.914 (95% CI: 0.874– 0.947). With a cutoff value of 0.519, the sensitivity and specificity for discriminating PTB from controls were 0.81 and 0.871, respectively. The nomogram also showed good calibration with the Hosmer–Lemeshow test (P=0.58) and good clinical practicality displayed by the decision curve analysis.Conclusion: A nomogram composed of IGRA, percentage of neutrophils, and expression of CD64, GZMA, and PRDM1 was established. The nomogram demonstrated a sensitivity and specificity of 81% and 87%, respectively, for differentiating TB from controls.Keywords: tuberculosis, prediction model, peripheral blood, interferon gamma release assay, transcripts of gene

Abstract Whole genome sequencing (WGS) has become a gold standard for diagnosing genomic variation. Peripheral blood is a common sample source for the extraction of nucleic acids for Next-Generation Sequencing (NGS) applications. Here, we present an integrated and fully automated device design that uses new concepts of fluid mechanics, heat-mass transfer, and thermodynamics of enzymatic reactions to extract nucleic acids from the blood and perform DNA library preparation from a pre-filled plate. We demonstrate that the presented device effectively extracts dsDNA with an average of 25.03 µg/mL and 25.91 µg/mL yield from citrate-stabilized human peripheral blood stored in Fresh (4 °C) and Frozen (-20 °C) conditions, respectively. Furthermore, our method automatically extracts nucleic acids and creates a high-quality sequence-ready DNA library from blood stabilized with citrate and EDTA for 8 samples simultaneously in a single run with a total operation time of ~ 7 h. Our results show the required coverage and depth of the genome, highlighting an essential application of this device in processing blood samples for genome sequencing. Graphical Abstract

Objective: The aim of this study was to assess the clinical value of metagenomic next-generation sequencing (mNGS) of blood samples for the identification of disseminated tuberculosis (DTB). Methods: A total of 48 individuals suspected of DTB were enrolled. All patients underwent mNGS of peripheral blood and conventional microbiological tests. Patient characteristics were collected from their medical records. Results: A total of 28 patients were diagnosed with DTB, whereas 20 patients were confirmed as non-DTB cases. In the DTB groups, 19 (67.9%) contained TB sequences, with specific reads of TB ranging from 1 to 219. The TB sequence was more detectable by mNGS in male patients, those with elevated PCT levels, those who are HIV positive, and those with a decreased CD4 T-cell count. The HIV-positive group shows higher TB mNGS reads (p = 0.012) and TB mNGS sensitivity (p = 0.05). The sensitivity of TB mNGS in blood samples was 80% for HIV-infected patients and 44.4% for non-HIV-infected individuals (p = 0.05). The non-HIV group had a higher prevalence of miliary tuberculosis (p = 0.018), and extrapulmonary tuberculosis was more prevalent in the HIV-positive group. Conclusion: Our research has shown that the mNGS of blood samples has excellent sensitivity for the diagnosis of DTB. The TB sequence was more detectable by mNGS in patients with elevated PCT levels, those who are HIV positive, and those with a decreased CD4 T-cell count. [ABSTRACT FROM AUTHOR]

Multiple myeloma (MM) is a disease which remains incurable. One of the main reasons is a weakened immune system that allows MM cells to survive. Therefore, the current research is focused on the study of immune system imbalance in MM to find the most effective immunotherapy strategies. Aiming to identify the key points of immune failure in MM patients, we analysed peripheral lymphocytes subsets from MM patients (n = 57) at various stages of the disease course and healthy individuals (HI, n = 15) focusing on T, NK, iNKT, B cells and NK-cell cytokines. Our analysis revealed that MM patients exhibited immune alterations in all studied immune subsets. Compared to HI, MM patients had a significantly lower proportion of CD4 + T cells (19.55% vs. 40.85%; p < 0.001) and CD4 + iNKT cells (18.8% vs. 40%; p < 0.001), within B cells an increased proportion of CD21LCD38L subset (4.5% vs. 0.4%; p < 0.01) and decreased level of memory cells (unswitched 6.1% vs. 14.7%; p < 0.001 and switched 7.8% vs. 11.2%; NS), NK cells displaying signs of activation and exhaustion characterised by a more than 2-fold increase in SLAMF7 MFI (p < 0.001), decreased expression of NKG2D (MFI) and NKp46 (%) on CD16 + 56 + and CD16 + 56- subset respectively (p < 0.05), Effective immunotherapy needs to consider these immune defects and monitoring of the immune status of MM patients is essential to define better interventions in the future. [ABSTRACT FROM AUTHOR]

Postmortem drug analysis is crucial in identifying the potential cause and manner of death. However, it is threatened by a significant phenomenon called postmortem redistribution (PMR), which refers to the alterations in drug levels occurring after death. This review aims to describe the PMR phenomenon, the mechanisms involved in the PMR of drugs, the various methods used to predict it, and various artifacts of postmortem drug concentrations. Several mechanisms, including passive diffusion from solid organs that act as drug reservoirs to the surrounding tissues, cadaveric changes after death (e.g., cell death, blood coagulation, hypostasis, and movements), and the putrefactive process, can result in artifacts of postmortem drug concentrations. The drug's chemical and pharmacokinetic properties (such as acidic/basic properties, lipophilicity, protein binding, high volume of distribution, and residual metabolic activity) are additional factors. Several markers, including cardiac blood-to-peripheral blood ratio (C/P), liver-to-peripheral blood ratio (L/P), amino acid markers such as methionine, quantitative structure–activity relationship (QSAR) approach, and F factor, have been proposed for interpreting the liability of drugs to PMR. Several artifacts may affect the reliability of postmortem drug analysis. Peripheral blood is preferred for postmortem drug sample collection. Numerous laboratories evaluate the redistribution potential of drugs after death using the C/P concentration ratio. Nevertheless, the L/P concentration ratio is proposed to be a more reliable marker for PMR determination. [ABSTRACT FROM AUTHOR]

Objective: To investigate the clinical value of EGFR mutation testing in peripheral blood of patients with non-small cell lung cancer. Method: This was a retrospective study. A total of 89 patients with non-small cell lung cancer (NSCLC) admitted to Affiliated Hospital of Hebei University between June 2019 and May 2023 were selected, and divided into the wild-type group and the mutant group according to the results of EGFR mutation testing in peripheral blood. Clinic pathological data of patients were collected and compared between the two groups, the correlation between EGFR mutation and the prognosis was analyzed. Result: No statistically significant difference in the mutation rate of EGFR gene was observed between peripheral blood ctDNA and tumor tissue (p=0.879). EGFR mutation in peripheral blood ctDNA was not significantly correlated with sex, age, ethnicity and ECOG score (p>0.05). The EGFR mutation rate was increased in patients with adenocarcinoma compared with that in patient with squamous carcinoma, and in non-smoking patients compared with that in smoking patients, and the differences were statistically significant (p<0.05). The 1-, 2-, and 3-year disease-free survival (DFS) rates were significantly increased in the mutant group compared with those in the wild-type group, respectively (p<0.05). Conclusion: EGFR mutation in peripheral blood and tumor tissues is highly consistent in NSCLC patients. The EGFR mutation rate is higher in adenocarcinoma and non-smoking patients, and the prognosis is better in patients with EGFR mutations. [ABSTRACT FROM AUTHOR]

Acute anterior uveitis (AAU) can cause great pain to patients. Th cytokines in peripheral blood are significantly changed in these patients, including serum interleukin-23 (IL-23), interleukin-17 (IL-17), interleukin-4 (IL-4), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) increase. However, the relationship between Th cytokines and recurrence of AAU is still unclear. Ninety-two cases of AAU were enrolled from January 2020 to April 2022 in our hospital (observation group). Levels of peripheral blood Th cytokines were detected, which were compared between acute and remission stages. After 6 months of follow-up, the relationships between Th cytokines in peripheral blood and recurrence in the observation group were analyzed. The prediction of Th cytokines on recurrence was analyzed. (1) The recurrence rate was 25.00%, and there were no statistical differences in the serum IL-23, IL-17, IL-4, IFN-γ, TNF-α and TGF-β 1 levels between patients with bilateral disease and unilateral disease (P < 0.05). (2) The serum IL-23, IL-17, IL-4, IFN-γ, TNF-α and TGF-β 1 levels of recurrence patients were higher than those of non-recurrence (t = 2.971, 5.357, 2.197, 2.766, 4.395, 2.983, P < 0.05). (3) The serum IL-23, IL-17, TNF-α levels were risk factors for recurrence (OR = 1.035, 1.210, 1.155, P < 0.05). (4) There were positive relationships between serum IL-23, IL-17, IL-4, IFN-γ, TNF-α and TGF-β 1 levels and recurrence (r = 0.317, 0.526, 0.248, 0.304, 0.480, 0.325, P < 0.05). [ABSTRACT FROM AUTHOR]

Peripheral blood involvement by MF/SS has significant implications for prognosis and treatment. Flow cytometry is commonly used to assess MF/SS by analyzing the ratio of CD26- and/or CD7-CD4 + T cells and assessment of immunophenotypic abnormalities. However, distinguishing normal from abnormal cells is not always easy. In this study, we aimed to establish quantitative thresholds to better distinguish normal CD4 + T cells from neoplastic CD4 + T cells. A retrospective analysis of flow cytometry data was performed on 30 MF/SS patients with a detectable abnormal T cell population (positive), 63 patients with suspected or confirmed cutaneous involvement without a detectable abnormal T cell population (negative), and 60 healthy controls (control). CD3 and CD4 median fluorescence intensity (MFI) was normalized to internal control subsets. Among the positive cases, 50% had CD3 expression outside ± 2 SD from the mean of the negative and control group in the CD4 + CD26- subset. The corresponding specificity of this threshold was 94%. The ± 2 SD threshold showed a sensitivity of 57% and a specificity of 94% for the CD3 intensity among the CD7-negative subset. For CD4 intensity, the ± 2 SD threshold had a sensitivity of 33.3% and specificity of 95% for the CD26-negative subset and a sensitivity of 37% and specificity of 95% for the CD7-negative subset. In our study, although changes in CD3 and CD4 intensity greater than ± 2 SD were specific for MF/SS, more subtle differences in the intensity of CD3 and CD4 should not be used as the sole abnormality to make a diagnosis of circulating MF/SS. [ABSTRACT FROM AUTHOR]

ABSTRACT Purpose Methods Results Conclusion Our study aimed to comprehensively describe the features of peripheral blood multiple immune cell phenotypes in solid tumor patients during pretreatment and after immunotherapy, providing a more convenient approach for studying the prognosis of immunotherapy in different solid tumor patients.We prospectively recruited patients with advanced solid tumors from Peking Union Medical College Hospital (PUMCH) between February 2023 and April 2024. Using multicolor flow cytometry, our study comprehensively observed and described the signatures of peripheral blood lymphocyte subsets including activation, proliferation, function, naïve memory, and T cell exhaustion immune cell subsets in this population of pretreatment and after immunotherapy.Our study enrolled 59 advanced solid tumor patients with immunotherapy and 59 healthy controls were matched by age and gender. The results demonstrated a marked upregulation in the expression of lymphocyte activation markers CD38 and HLA‐DR, as well as exhaustion and proliferation markers PD‐1 and Ki67, in solid tumor patients compared to healthy controls. After immune checkpoint blockade (ICB) treatment, mainly the expression of Ki67CD4+T and HLA‐DRCD38CD4+T, was significantly upregulated compared to pretreatment levels (p = 0.017, p = 0.019, respectively). We further found that gynecological tumors with better prognoses had higher baseline activation levels of CD4+ T cells compared to other solid tumors with poorer prognoses.Our study elucidated the characteristics of different lymphocyte subsets in the peripheral blood of solid tumor patients. Further research revealed changes in the phenotypes of different lymphocyte subsets after ICIs treatment, with the activated phenotype of CD4+ T cells playing a crucial role in the antitumor effect. This lays the groundwork for further exploration of prognostic biomarkers and predictive models for cancer patients with immunotherapy. [ABSTRACT FROM AUTHOR]

BACKGROUND: In recent years, there have been many studies on the mechanism of exosomal non-coding RNA in gestational diabetes mellitus, but there is a lack of the latest systematic review of exosomes from different sources, especially placental sources. OBJECTIVE: To summarize the changes and potential roles of microRNA (miRNA), long non-coding RNA (lncRNA), circular RNA (circRNA), and exosomes in gestational diabetes mellitus to provide potential targets for early screening and treatment of clinical gestational diabetes mellitus. METHODS: A literature search was conducted on PubMed, Web of Science, China National Knowledge Infrastructure, WanFang Data, and VIP databases to retrieve relevant articles on non-coding RNA or exosomal non-coding RNA in relation to gestational diabetes mellitus. A total of 74 articles were included for review. RESULTS AND CONCLUSION: (1) Non-coding RNAs play important pathological and physiological roles in the lifecycle activities, and increasing evidences suggest that non-coding RNAs are involved in the occurrence and development of gestational diabetes mellitus by regulating various physiological functions. This provides a new direction for the research of gestational diabetes mellitus. (2) Exosomes are widely present in the human body. Various cells can secrete exosomes, such as red blood cells, epithelial cells, and placental cells. Non-coding RNAs found in exosomes from different sources have been demonstrated to play a role in the pathogenesis, diagnosis, and treatment of gestational diabetes mellitus. (3) MiRNA and gestational diabetes mellitus: The role of peripheral blood miRNA in gestational diabetes mellitus is mainly to affect the functions of trophoblast cells, pancreatic beta cells and blood glucose levels in gestational diabetes mellitus; placental miRNA can reflect the severity of gestational diabetes and impair the function of trophoblast cells. (4) LncRNA and gestational diabetes mellitus: Peripheral blood lncRNA can induce insulin resistance through the phosphatidylinositol 3-kinase/protein kinase B pathway and may provide new insights for the diagnosis and treatment of gestational diabetes mellitus; placental lncRNA can regulate proliferation and migration of placental trophoblast cells, promoting the occurrence and development of gestational diabetes mellitus. (5) CircRNA and gestational diabetes mellitus: Peripheral blood and placental circRNA can induce the occurrence and development of gestational diabetes mellitus by impairing the proliferation, migration and metabolism of placental trophoblast cells. (6) Non-coding RNA in exosomes and gestational diabetes mellitus: Peripheral blood non-coding RNA in exosomes can affect gestational diabetes mellitus blood glucose levels and glucose homeostasis, and participate in the occurrence and development of gestational diabetes mellitus by influencing placental function. (7) Non-coding RNA has the potential to serve as biomarkers for early diagnosis of gestational diabetes mellitus. Additionally, engineered exosomes can better achieve targeted therapy for gestational diabetes mellitus. These latest findings provide a reference for both basic research and clinical translation of gestational diabetes mellitus. (8) In the future, improvements in the extraction and purification methods of peripheral blood exosomes should be improved, and factors such as race, diet and physical activity should be excluded to improve the reproducibility of results. Further prospective clinical studies are required to explore the clinical application of circulating non-coding RNA and exosomes in the prediction and diagnosis of gestational diabetes mellitus. [ABSTRACT FROM AUTHOR]

Treatments targeting the immune system only benefit a subset of patients with bladder cancer (BC). Biomarkers predictive of BC progression and response to specific therapeutic interventions are required. We evaluated whether peripheral blood immune subsets and expression of clinically relevant immune checkpoint markers are associated with clinicopathologic features of BC. Peripheral blood mononuclear cells isolated from blood collected from 23 patients with BC and 9 age‐matched unaffected‐by‐cancer control donors were assessed using a 21‐parameter flow cytometry panel composed of markers of T, B, natural killer and myeloid populations and immune checkpoint markers. Patients with BC had significantly lower numbers of circulating CD19+ B cells and elevated circulating CD4+CD8+ T cells compared with the control cohort. Immune checkpoint markers programmed cell death protein 1 (PD‐1) and T‐cell immunoglobulin and mucin‐domain containing‐3 (TIM‐3) were elevated in the total peripheral immune cell population in patients with BC. Within the BC cohort, PD‐1 expression in T and myeloid cells was elevated in muscle‐invasive compared with non–muscle‐invasive disease. In addition, elevated T, B and myeloid PD‐1 cell surface expression was significantly associated with tumor stage, suggesting that measures of peripheral immune cell exhaustion may be a predictor of tumor progression in BC. Finally, positive correlations between expression levels of the various immune checkpoints both overall and within key peripheral blood immune subsets collected from patients with BC were observed, highlighting likely coregulation of peripheral immune checkpoint expression. The peripheral blood immunophenotype in patients with BC is altered compared with cancer‐free individuals. Understanding this dysregulated immune profile will contribute to the identification of diagnostic and prognostic indicators to guide effective immune‐targeted, personalized treatments. [ABSTRACT FROM AUTHOR]

BACKGROUND: Allergic Rhinitis (AR), a prevalent condition in otorhinolaryngology, is mediated by Type 1 hypersensitivity through IgE, characterized by Type 2 inflammatory response and eosinophil infiltration in the nasal mucosa. Since AR disease exhibits significant heterogeneity in symptom severity, an objective assessment of AR severity may facilitate better individualized treatment. OBJECTIVE: To explore the changes in peripheral blood IL-9, Th9, and BAFF levels of allergic rhinitis (AR) in patients and the clinical significance associated with it. METHODS: A retrospective study selected 80 AR patients admitted from January 2022 to October 2022 as the case group, dividing them into mild and moderate-to-severe groups based on symptom scores. Concurrently, 50 patients without AR, who were treated for nasal bone fractures or underwent septoplasty, were selected as the group for comparison. Alterations in the expression levels of peripheral blood IL-9, Th9, and BAFF were analyzed and compared among the different groups. The diagnostic value of serum BAFF for the severity of AR was analyzed using the receiver operating characteristic (ROC) curve. RESULTS: Noticeable variations were observed in clinical variables among the three groups such as, total IgE levels, peripheral blood eosinophil count and proportion, TNSS, and VAS (P < 0.05), while no statistically significant differences were observed in other variables (P > 0.05). The comparison of IL-9, Th9, and BAFF among the three groups revealed statistically significant differences (P < 0.05). Analysis using multivariate logistic regression revealed that IL-9 (OR = 2.365), Th9 (OR = 2.186), BAFF (OR = 2.307) were influencing factors of moderate-to-severe AR (P < 0.05). The ROC curve indicated that the AUC for the diagnosis of moderate-to-severe AR by IL-9, Th9, BAFF were 0.770, 0.734, 0.761, respectively, and the combined detection AUC was 0.888, an area under the curve higher than individual testing. CONCLUSION: Changes in peripheral blood IL-9, Th9, and BAFF levels in AR patients may function as indicators to assess the level of severity in diagnostic procedures. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of neutrophil/lymphocyte ratio, platelet/lymphocyte ratio, and fibrin/albumin ratio in systemic inflammatory markers on bone loss and normal bone function after fractures.Methods A retrospective study was conducted from January 2023 to September 2023 on the medical records of 2 652postoperative fracture patients publicly available in the electronic case system of the 1st Affiliated Hospital of Kunming Medical University. According to the results of bone density testing, patients were divided into two groups:141 patients with the reduced bone mass and 79 patients with the normal bone mass. According to the collected data, the relevant information of each group, including NLR, PLR, FAR, gender, age, triglycerides, blood glucose, blood pressure, etc, was analyzed using SPSS 25.0 software. Results There was no significant statistical difference in NLR, PLR, and FAR between the two groups (P>0.05). There were statistical differences in BMI, age and gender (P<0.05). Conclusion The inflammatory response of bone loss after the fracture may be different from the inflammatory factors of female osteoporosis. [ABSTRACT FROM AUTHOR]

Background and Objectives: Pregnancy loss can occur in different stages of pregnancy for various reasons; however, the increase in embryonic mortality in the early stages of pregnancy is currently a concern in the dairy cow industry. Granulocyte-Colony Stimulating Factor (G-CSF) is a hematopoietic cytokine that is mainly produced by monocytes and macrophages and causes the proliferation and differentiation of myeloid hematopoietic cells. Interferon-Stimulated Gene-15 (ISG-15) functions as an intracellular ubiquitin homologue and extracellular cytokine that may serve as a surrogate marker for early pregnancy in ruminants. It is assumed that these cytokines increase in the peripheral blood simultaneously with the production of interferons in the uterus. The present study was an attempt to shorten the number of open days in dairy cows by detecting pregnancy in the early stages of pregnancy using cytokines such as G-CSF and ISG-15. Therefore, this study aimed to compare the gene expression changes between pregnant and non-pregnant dairy cows with early or late embryonic death, so that open days can be reduced in dairy cows. Material and Methods: Blood samples from 30 animals with embryonic death (10 cows for each time group) and 30 cows with normal pregnancy (30 cows for each time group) were collected in vacuum tubes containing EDTA anticoagulant on days 15–24, 25– 34, and 35–44 after inoculation. Cows were divided into pregnant and non-pregnant groups based on progesterone concentration on day 21, and non-pregnant cows were classified into those with early embryonic death (day 1-26) and late embryonic death (day 26-45). Neutrophils were then extracted from the blood samples, and G-CSF and ISG-15 gene expression was measured using Real-Time PCR. Results: The results showed that the concentration of progesterone in the blood plasma of pregnant cows was significantly higher than that of non-pregnant cows in the time interval of 15 to 24, 25 to 34 and 35 to 44 days after artificial insemination. The concentration of glucose in the blood plasma of pregnant cows between 15 and 24 h (P<0.001) was significantly higher in the time intervals of 25 to 34 days and 35 to 44 days, compared to the concentration of glucose in the blood plasma of non-pregnant cows. Cholesterol concentration in the plasma of pregnant cows between 25 and 34 days after artificial insemination (AI) showed a statistically significant difference (P<0.05) from the cholesterol concentration in the blood plasma of non-pregnant cows. Examination of the concentration of triglycerides in the blood plasma of pregnant cows at 15–24, 25–34, and 35–44 days after inoculation showed no statistically significant difference from the concentration of triglycerides in the blood plasma of non-pregnant cows on the same days. The highest level of G-CSF gene expression was observed between days 15 and 24 in the blood neutrophils of pregnant cows. In addition, the highest ISG-15 expression was observed between 35 and 44 days after inoculation. Conclusion: The results of the present study showed that these genes can be used clinically to diagnose early and late embryonic death. [ABSTRACT FROM AUTHOR]

The pathology of respiratory syncytial virus (RSV) infection remains unclear. An unbalanced immune response to RSV infection can lead to immunopathology, causing airway damage and impaired exchange of oxygen and carbon dioxide between the air and the bloodstream. We aimed to evaluate the association of the expression of inflammatory and antiviral genes in peripheral blood with severe hypoxemia in children with RSV infection seen in the hospital emergency room. We conducted a cross-sectional study on 121 RSV-infected children seen in hospital emergency rooms between 2015 and 2023. Total RNA was extracted from whole blood samples, and gene expression (IL-6, TNFa, CXCL8, ISG15, IFIT1, RIGI, IFNb, CCL5, and CXCL10) was quantified using quantitative RT-PCR. The outcome variable was having severe hypoxemia (SpO2 ≤ 90%). The association analysis was performed using a volcano plot, adjusted logistic regression, and orthogonal partial least squares discriminant analysis (OPLSDA). We found that 26 of 121 children had severe hypoxemia (SpO2 ≤ 90%). CXCL8 was overexpressed [fold changes (FC) > 2; q-value < 0.05], and ISG15, IFIT1, RIGI, IFNb, CCL5, and CXCL10 were underexpressed (FC <0.5; q-value <0.05) in children with severe hypoxemia. These associations were ratified using adjusted logistic regression. The OPLS-DA showed that the gene expressions of CXCL8, ISG15, IFIT1, RIGI, and CXCL10 had values of variable importance in projection (VIP) ≥1, being the most relevant features. In conclusion, an imbalance favoring inflammation over antiviral defense may contribute to the pathogenesis of severe hypoxemia in RSV-infected children. These findings provide valuable insights into the pathology of RSV infection. [ABSTRACT FROM AUTHOR]

Whole genome sequencing (WGS) has become a gold standard for diagnosing genomic variation. Peripheral blood is a common sample source for the extraction of nucleic acids for Next-Generation Sequencing (NGS) applications. Here, we present an integrated and fully automated device design that uses new concepts of fluid mechanics, heat-mass transfer, and thermodynamics of enzymatic reactions to extract nucleic acids from the blood and perform DNA library preparation from a pre-filled plate. We demonstrate that the presented device effectively extracts dsDNA with an average of 25.03 µg/mL and 25.91 µg/mL yield from citrate-stabilized human peripheral blood stored in Fresh (4 °C) and Frozen (-20 °C) conditions, respectively. Furthermore, our method automatically extracts nucleic acids and creates a high-quality sequence-ready DNA library from blood stabilized with citrate and EDTA for 8 samples simultaneously in a single run with a total operation time of ~ 7 h. Our results show the required coverage and depth of the genome, highlighting an essential application of this device in processing blood samples for genome sequencing. [ABSTRACT FROM AUTHOR]

Parkinson's disease (PD) is one of the most common progressive neurodegenerative diseases. An important feature of the disease is its long latent period, which necessitates search for prognostic biomarkers. One method of identifying biomarkers of PD is to study changes in gene expression in peripheral blood of the patients in early stages of the disease and have not been treated. In this study, we analyzed relative mRNA levels of the genes GRIPAP1, DLG4, KIF1B, NGFRAP1, and NRF1, which are associated with neurotransmitter transport, apoptosis, and mitochondrial dysfunction, in the peripheral blood of PD patients using reverse transcription and real-time PCR with TaqMan probes. The results of this study suggest that the GRIPAP1 and DLG4 genes could be considered as potential biomarkers for the early clinical stages of Parkinson's disease. The data obtained may indicate that NGFRAP1 is involved in pathogenesis of both PD and other neurodegenerative diseases. Furthermore, in the early clinical stages of the disease we studied, the KIF1B and NRF1 genes were found not to be involved in PD pathogenesis at the expression level. [ABSTRACT FROM AUTHOR]

Thrombosis, a prevalent condition, can provoke severe health issues like acute coronary syndrome (ACS), deep vein thrombosis (DVT), and pulmonary embolism (PE). The rising incidence of these diseases annually significantly impacts patient wellbeing and poses a substantial burden on healthcare systems. Recombinant neorudin is a developing anticoagulant drug for thrombotic diseases whose phase I clinical trials has been completed. The distribution pattern of it and its active metabolite, hirudin, in thrombi, blood surrounding the thrombus and peripheral blood remains uncertain. This study explored their distribution using a rat arteriovenous bypass thrombosis model, revealing higher neorudin levels in blood surrounding the thrombus and elevated hirudin concentrations in thrombus. Recombinant neorudin significantly increased Thrombin Time (TT) in both plasma surrounding the thrombus and peripheral blood, and reduced the wet weight of the thrombus. The results above demonstrated the anticoagulant and antithrombotic efficacy of recombinant neorudin in vivo. Give the distribution pattern of neorudin and hirudin, we hypothesized that neorudin was cleaved at the site of thrombus formation to produce hirudin, leading to the rapid accumulation of hirudin within local thrombi and resulting in a higher concentration inside the thrombus. This insight was crucial for understanding the action mechanisms of anticoagulants in thrombosis management and provided a valuable guidance for therapeutic strategies in treating thrombotic diseases. [ABSTRACT FROM AUTHOR]

Blood lymphocyte reference ranges in non-pregnant females are established, but changes in pregnancy are less well understood. The early identification of immunological markers that could suggest an increased risk of early pregnancy loss may allow for timely intervention to improve outcomes. A lymphocytic immunophenotype provides a broad assessment of important immune parameters and potential indicators, which may be of relevance to pregnancy outcome. Comparison of immunophenotype results on the day of a positive hCG after embryo transfer between successful and failed pregnancies allows for this assessment. Baseline non-pregnant lymphocyte percentage and cell/µL profiles were established with a comprehensive panel on 93 age-matched male factor controls. Sixty-five in-vitro fertilisation (IVF) patients had an immunophenotype assessment on the day of a positive hCG, followed by further hCG tests and ultrasound monitoring as required to ultimately evaluate success (live birth) or failure (miscarriage). Thirty-one pregnancies were viable, leading to a live birth, while 34 ended in miscarriage. Total CD56, pNK, NKT, CD4 and CD8 levels were equivalent between all groups. Regardless of the outcome, B lymphocytes increased in pregnancy compared to controls. Of interest, in the later miscarriage cohort, pNK-specific CD69 was reduced (1.6 vs 5.4%, P = 0.02), while CD57+ cells were increased (45.4 vs 38.9%, P = 0.025). Corresponding changes were observed in cell/µL concentrations. Low level CD69 activation and elevated CD56dim and CD57+ NK cells were identified as markers that could potentially identify a pregnancy at risk of miscarriage, with further study needed to explore whether these changes represent cause or effect.

Pelvic organ prolapse (POP) is a common gynecological disease caused by pathological defects, lesions, or mechanical weakening of the support structures of the pelvic floor. However, its pathogenesis is unclear. Circular RNAs (circRNAs) are covalently closed, endogenous biomolecules, which are thought to play an important role on skeletal muscle development by regulating gene expression. In this study, five pairs of peripheral blood samples from control and POP groups were used for circRNA sequencing analysis to obtain differential expression profiles. A total of 75 differentially expressed circRNAs (DEcircRNAs) were identified (fold change >2.0, P < 0.05). Furthermore, RT-qPCR confirmed that the expression levels of two circRNAs (hsa_circ_0067962 and hsa_circ_0057051) were significantly lower in the POP group. The two validated DEcircRNAs were abundantly involved in the collagen catabolic process. The circRNA-miRNA-mRNA network of two DEcircRNAs comprised nine mRNAs, which indicated that hsa_circ_0067962 and hsa_circ_0057051 may be involved in the pathogenesis of POP by regulating these nine mRNAs.

BackgroundPancreatic cancer remains one of the deadliest malignancies, largely due to its late diagnosis and lack of effective therapeutic targets.Materials and methodsUsing traditional machine learning methods, including random-effects meta-analysis and forward-search optimization, we developed a robust signature validated across 14 publicly available datasets, achieving a summary AUC of 0.99 in training datasets and 0.89 in external validation datasets. To further validate its clinical relevance, we analyzed 55 peripheral blood samples from pancreatic cancer patients and healthy controls using qPCR.ResultsThis study identifies and validates a novel five-gene transcriptomic signature (LAMC2, TSPAN1, MYO1E, MYOF, and SULF1) as both diagnostic biomarkers and potential drug targets for pancreatic cancer. The differential expression of these genes was confirmed, demonstrating their utility in distinguishing cancer from normal conditions with an AUC of 0.83. These findings establish the five-gene signature as a promising tool for both early, non-invasive diagnostics and the identification of actionable drug targets.ConclusionA five-gene signature is established robustly and has utility in diagnostics and therapeutic targeting. These findings lay a foundation for developing diagnostic tests and targeted therapies, potentially offering a pathway toward improved outcomes in pancreatic cancer management.

Wei Hong,1 Lei Zhang,1 Youkun Qi,2 Yanjun Wang,1 Wentao Wang3 1Oncology, The Second People’s Hospital of Liaocheng, Liaocheng, Shandong, People’s Republic of China; 2Pharmacy, The Second People’s Hospital of Liaocheng, Liaocheng, Shandong, People’s Republic of China; 3Critical Care Medicine, The Second People’s Hospital of Liaocheng, Liaocheng, Shandong, People’s Republic of ChinaCorrespondence: Wentao Wang, Critical Care Medicine, The Second People’s Hospital of Liaocheng, Liaocheng, Shandong, People’s Republic of China, Email 287474031@qq.comPurpose: Lung cancer remains a leading cause of cancer-related death and chemotherapy stands as a fundamental component in therapy. Chemotherapy-induced myelosuppression encompasses a spectrum of hematological declines, including not only neutrophils but also lymphocytes, hemoglobin levels and platelets. This retrospective cohort study investigates alterations in peripheral blood lymphocyte subsets. By uncovering these changes, our goal is to refine patient management strategies, ensuring that the benefits of chemotherapy are maximized while minimizing its detrimental effects.Patients and Methods: We retrospectively analyzed 159 lung cancer patients. Patients were categorized as “NT” (n=108, no previous anti-tumor therapy), and “PT” (n=51, prior therapy followed by at least a two-month treatment-free interval). Post-chemotherapy, patients were reassessed and grouped into “EarlyCycle” for those who underwent four or fewer cycles, and “LateCycle” for those who underwent more than four cycles.Results: The study focused on analyzing the percentages of lymphocyte subsets, including T cells (CD4+, CD8+), B cells, and natural killer (NK) cells, across these groups. For T cells, the EarlyCycle group exhibited a significant increase compared to NT (0.7783 vs 0.7271; p=0.0017) and PT (0.7783 vs 0.6804; p=1.6e-05). B cells showed a significant decrease from NT to LateCycle (0.1014 vs 0.0817; p=2.2e-05) and from PT to LateCycle (0.1317 vs 0.0817; p=6.2e-10). NK cells significantly decreased in the EarlyCycle group compared to NT (0.1109 vs 0.1462; p=0.00816) and PT (0.1109 vs 0.1513; p=0.00992), with no significant change in the LateCycle group compared to either NT or PT (p> 0.05).Conclusion: Chemotherapy significantly affects lymphocyte subsets in a treatment-specific manner. The EarlyCycle group experienced a reduction in NK cell and an increase in T cell, suggesting a damage of innate immunity and an early shift towards adaptive immunity. The LateCycle group showed a substantial decrease in B cell, indicating a delayed effect on humoral immunity components.Keywords: chemotherapy, lung cancer, myelosuppression, peripheral blood, lymphocyte subsets

Abstract Purpose This study aimed to investigate the clinical significance of combining peripheral blood miR-21 and miR-486 with CT for the early cancer diagnosis in pulmonary nodules. Methods A total of 215 patients diagnosed with isolated pulmonary nodules with a history of smoking were selected as researchsubjects. 30 healthy volunteers with a history of smoking were recruitedas the control group.The selection of subjectswas based on the presence of isolated pulmonary nodules detected on chest CT scans. The training set consisted of 65 patients with lung nodules and 30 healthy smokers, while the verification setincluded 150 patients with lung nodules. Results Compared with the control group, the plasma expression level of miR-210 was significantly higher in the group of patients with benign pulmonary nodules (P

Endah Wulandari,1 Rr Ayu Fitri Hapsari2 1Department of Biochemistry, Islamic State University Syarif Hidayatullah Jakarta of Medicine Faculty, South Tangerang, Banten, Indonesia; 2Department of Histology, Islamic State University Syarif Hidayatullah Jakarta of Medicine Faculty, South Tangerang, Banten, IndonesiaCorrespondence: Endah Wulandari, Department of Biochemistry, Islamic State University Syarif Hidayatullah Jakarta of Medicine Faculty, South Tangerang, Banten, Indonesia, Email endah.wulandari@uinjkt.ac.idPurpose: The main aim of this study was to compare and analyze hematological profiles using menstrual blood, as an alternative to peripheral blood.Patients and Methods: This study used menstrual and peripheral blood samples from women who were menstruating. The design of this research is analytical observational.Results: Menstrual blood can show an overall hematological profile similar to peripheral blood. Data shows the detection of blood component parameters, white blood cells and reticulocytes in MB with a range within and outside normal blood. Data on MB that show higher values (WBC, MCH, MCHC, PLT, RDW-CV, PDW, MPV, P-LCR, PCT, neutrophils, lymphocytes, monocytes, basophils, reticulocytes, LFR, Ret-He) and lower values lower (RBC, HGB, HCT, MVC, RDW-SD, Eosinophils, IRF, MFR, HFR) when compared with peripheral blood controls. The hematological profiles of Menstrual and peripheral blood showed significant differences (p < 0.01) for several parameters, while several other parameters did not show significant differences (p > 0.05) according to the Wilcoxon test.Conclusion: All hematological profile parameters were detected in menstrual blood. The new concept that menstrual blood can be used as a supporting medium for hematological examinations opens up opportunities for developing independent hematological detection tools in productive women.Keywords: hematological, menstrual blood, peripheral blood, WBC, RBC, reticulocyte

Abstract Background Bone marrow (BM) evaluation is the de facto standard for diagnosis, molecular analysis, risk stratification, and therapy response assessment in acute myeloid leukemia (AML), but in patients with a high number of circulating blast cells, the peripheral blood (PB) sample could provide similar information as BM. However, there is no large‐scale molecular study comparing the two specimens in terms of their gene expression profiles, cellular heterogeneities, and ex‐vivo drug sensitivity. Methodology We used (i) the BEAT‐AML cohort each with detailed molecular data; (ii) cell‐type deconvolution to estimate leukemic and immune cell proportions between specimen types; (iii) differential expression (DE) and drug‐cell type association analysis; and (iv) logistic regression models to assess the association between induction therapy response, cell‐type composition and first‐line drug treatment. Results Results: We identified 207 patients having BM and 116 patients having PB samples. There was a total of 1271 DE genes (false discovery rate

Introduction: Endometriosis (EMs) and adenomyosis (AD) are common gynecological diseases that impact women's health, and they share symptoms such as dysmenorrhea, chronic pain, and infertility, which adversely affect women's quality of life. Current diagnostic approaches for EMs and AD involve invasive surgical procedures, and thus, methods of noninvasive differentiation between EMs and AD are needed. This retrospective cohort study introduces a novel, noninvasive classification methodology employing a stacked ensemble machine learning (ML) model that utilizes peripheral blood and coagulation markers to distinguish between EMs and AD. Methods: The study included a total of 558 patients (329 with EMs and 229 with AD), in whom key hematological and coagulation markers were analyzed to identify distinctive profiles. Feature selection was conducted through ML (logistic regression, support vector machine, and K-nearest neighbors) to determine significant hematological markers. Results: Red cell distribution width, mean corpuscular hemoglobin concentration, activated partial thromboplastin time, international normalized ratio, and antithrombin III were proved to be the key distinguishing indexes for disease differentiation. Among all the ML classification models developed, the stacked ensemble model demonstrated superior performance (area under the curve = 0.803, 95% credibility interval = 0.701-0.904). Our findings demonstrate the effectiveness of the stacked ensemble ML model for classifying EMs and AD. Discussion: Integrating biomarkers into this multi-algorithm framework offers a novel approach to noninvasive diagnosis. These results advocate for the application of stacked ensemble ML utilizing cost-effective and readily available peripheral blood and coagulation indicators for the early, rapid, and noninvasive differential diagnosis of EMs and AD, offering a potentially transformative approach for clinical decision-making and personalized treatment strategies. [ABSTRACT FROM AUTHOR]

Endometriosis, a debilitating condition, affects one in ten women of reproductive age. Its pathophysiology remains unclear, though deficiencies in immune surveillance are thought to create an environment conducive to the evasion of ectopic endometrial cells from the immune system. Our research explores the immunological impact of endometriosis both locally and systemically, emphasizing natural killer (NK) and T cell subpopulations. We incorporated 62 female patients who underwent laparoscopic surgery; of those, 47 had endometriosis, and 15 were controls. We collected peritoneal fluid (PF) and peripheral blood (PB) samples which were tagged with monoclonal antibodies and subsequently scrutinized using flow cytometry. Our findings revealed significant differences in immunological profiles based on demographic factors and symptomatology. In the endometriosis cohort, there was an increase in PB CD56HiCD16dim and PF CD8+ CD56dimCD16Hi NK cells. CD16+ CD4 T cell levels were significantly lower in the PB of endometriosis patients who smoke. Individuals with more severe disease displayed significantly higher levels of PB CD16+ CD8 T cells, which also increased in those with non-menstrual pelvic pain. Dysmenorrhea severity correlated with a progressive increase in PF CD8+ CD56dimCD16Hi NK cells. These variations in specific lymphocyte subsets, namely, within NK and T cells, suggest potential immunological mechanisms in the evolution and clinical presentation of endometriosis. [ABSTRACT FROM AUTHOR]

Simple Summary: Peripheral blood analyses can offer a minimally invasive view into systemic immunity during cancer and can lead to the identification of biomarkers for cancer screening and therapeutic management. While a limited number of studies have reported blood transcriptome in breast cancer (BC) using RNA-seq analysis, our study is the first that aimed to identify potential BC biomarkers by analyzing transcriptome at an isoform level in peripheral blood mononuclear cells (PBMCs) from BC patients and healthy women. Our approach has led to the identification of an isoform of the RASGEF1A gene, the ENST00000374459 transcriptional variant, as a promising blood mRNA biomarker for distinguishing BC and healthy subjects. Additionally, our association analysis with clinicopathological characteristics revealed that lower ENST00000374459 expression in PBMCs of breast cancer patients was associated with higher proliferation and circulating tumor DNA (ctDNA) shedding, thereby linking expression of this isoform in blood immune cells to cancer progression and spreading. Background: Breast cancer (BC) comprises multiple subtypes with distinct molecular features, which differ in their interplay with host immunity, prognosis, and treatment. Non-invasive blood analyses can provide valuable insights into systemic immunity during cancer. The aim of this study was to analyze the expression of transcriptional isoforms in peripheral blood mononuclear cells (PBMCs) from BC patients and healthy women to identify potential BC immune biomarkers. Methods: RNA sequencing and isoform-level bioinformatics were performed on PBMCs from 12 triple-negative and 13 luminal A patients. Isoform expression validation by qRT-PCR and clinicopathological correlations were performed in a larger cohort (156 BC patients and 32 healthy women). Results: Transcriptional analyses showed a significant (p < 0.001) decrease in the ENST00000374459 RASGEF1A isoform in PBMCs of BC compared to healthy subjects, indicating disease-related expression changes. The decrease was associated with higher ctDNA and Ki-67 values. Conclusions: The levels of the RASGEF1A transcriptional isoform ENST00000374459 may have the potential to distinguish between BC and healthy subjects. The downregulation of ENST00000374459 in breast cancer is associated with higher proliferation and ctDNA shedding. Specialized bioinformatics analyses such as isoform analyses hold significant promise in the detection of biomarkers, since standard RNA sequencing analyses may overlook specific transcriptional changes that may be disease-associated and biologically important. [ABSTRACT FROM AUTHOR]

Objective: This study assessed the diagnostic value of microRNA-200 (miR-200) expression in peripheral blood-derived extracellular vesicles (EVs) in early-stage non-small cell lung cancer (NSCLC). Methods: This study retrospectively analyzed 100 healthy volunteers (the control group) receiving physical examinations, 168 early-stage NSCLC patients (the NSCLC group), and 128 patients with benign lung nodules (the benign group). The basic and clinical data of participants were obtained, including age, sex, smoking history, carbohydrate antigen 242 (CA242), carcinoembryonic antigen (CEA), carbohydrate antigen 199 (CA199), forced expiratory volume in 1 s, maximal voluntary ventilation, forced vital capacity, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and miR-200 expression. The correlation of miR-200 expression in peripheral blood-derived EVs with CA242, CEA, and CA199 was analyzed, and the diagnostic value of peripheral blood-derived EV miR-200 for early-stage NSCLC was assessed. The risk factors of early-stage NSCLC development were also determined. Results: Age, the percentage of patients with smoking history, CA242, CEA, CA199, IL-6, and TNF-α levels, and miR-200 expression in peripheral blood-derived EVs were significantly higher in the NSCLC group than in the benign and control groups. Lung disease patients with high miR-200 expression in peripheral blood-derived EVs comprised a higher percentage of patients with smoking history and mixed lesions and had higher CA242, CEA, CA199, and TNF-α levels than those with low miR-200 expression in peripheral blood-derived EVs. In lung diseases, miR-200 expression in peripheral blood-derived EVs was significantly and positively correlated with CA242, CEA, and CA199. Peripheral blood-derived EV miR-200 combined with CA242, CEA and CA199 had higher diagnostic value (area under the curve = 0.942) than single detection, along with higher specificity, and high expression of peripheral blood-derived EV miR-200 was an independent risk factor for early-stage NSCLC. Conclusion: Peripheral blood-derived EV miR-200 expression in patients with lung diseases is closely correlated with CA242, CEA, and CA199, and high expression of peripheral blood-derived EV miR-200 is an independent risk factor for early-stage NSCLC and is of high clinical diagnostic value for early-stage NSCLC. [ABSTRACT FROM AUTHOR]

This paper presents the evolution of Hematopoietic Stem Cell Transplants (HSCT) in Panama, as well as the main types of transplants and hematopoietic cell donors in the country. There are few research works on the subject. The study is of a mixed, concurrent nested type. Data from the Social Security Fund (CSS), the National Oncology Institute (ION) and the Panamanian Transplant Organization (OPT) were used. Among the main results, it was found that up to December 2022, more than 600 HSCT had been performed, with just over 250 of these being from the CSS and 300 from the ION, the majority of which were autologous. [ABSTRACT FROM AUTHOR]

Background: RUNX3 is hypermethylated in multiple cancers. TIMP2 also functions as a regulator of tumors. However, there are only very few reports on the association of methylation of RUNX3 and TIMP2 with lung cancer (LC) in peripheral blood. Methods: 426 LC patients and 428 age- and sex-matched healthy controls were recruited. DNA methylation in blood was semi-quantitively assessed by mass spectrometry. For the association analysis, binary logistic regression analysis adjusted covariant was applied, and ORs were presented as per +10% methylation. Results: Hypermethylation of CpG_1, CpG_5 and CpG_8 in RUNX3 was significantly associated with LC (ORs = 1.45, 1.35 and 1.35, respectively, adjusted p < 0.05), and even stage I LC. The association between the three RUNX3 CpG sites and LC was enhanced by increased age (> 55 years, ORs ranged from 1.43 to 1.75, adjusted p < 0.05), male gender (ORs ranged from 1.47 to 1.59, adjusted p < 0.05) and tumor stage (stage II&III&IV, ORs ranged from 1.86 to 3.03, adjusted p < 0.05). Conclusions: This study suggests a significant association between blood-based RUNX3 hypermethylation and LC, especially in elder people, in males and in LC patients with advanced stage. Clinical significance: The blood-based RUNX3 hypermethylation is associated with LC, especially stage I LC. LC-associated blood RUNX3 hypermethylation is increased with age. More LC-associated RUNX3 hypermethylation are found in males. [ABSTRACT FROM AUTHOR]

Recurrent pregnancy loss (RPL) is thought to be related to maternal-fetal immune tolerance disorders. Immune monitoring of RPL patients mainly involves two aspects: inflammatory factors and immune cells. However, most observational studies have reported controversial findings. This study aimed to confirm whether abnormal inflammatory factors and immune cells in peripheral blood may lead to RPL, and guide clinical immune monitoring. We demonstrated causality using two-sample Mendelian randomization. Sensitivity analysis, reverse Mendelian randomization and meta-analysis were used to enhance the effectiveness of the results. There was a causal relationship between the level of IL-12 (OR = 1.78, 95% CI = 1.25–2.55; P = 0.00149) and RPL for 41 inflammatory factors. We screened 5 groups of immune cell subtypes that were causally associated with RPL: switched memory B-cell absolute count (OR = 0.66, 95% CI = 0.49–0.87, P = 0.00406), IgD + CD24 + B-cell absolute count (OR = 0.69, 95% CI = 0.53–0.88, P = 0.00319), CD39 + resting CD4 regulatory T-cell %CD4 regulatory T-cell (OR = 0.86, 95% CI = 0.78–0.95, P = 0.00252), activated & resting CD4 regulatory T-cell %CD4 regulatory T-cell (OR = 0.89, 95% CI = 0.82–0.97, P = 0.00938) and CD45 RA + CD28-CD8 + T-cell %CD8 + T-cell (OR = 0.99, 95% CI = 0.98-1.00, P = 0.01231). In terms of inflammatory factors, a causal relationship between IL-12 and RPL in peripheral blood was confirmed. We also identified five immune cell phenotypes that play a protective role. This suggests that there may be protective B cells and CD8 + T-cell subsets in peripheral blood, and the protective effect of Tregs was proved again. Immune monitoring of peripheral blood in patients with RPL seems to be necessary and the foundation for precision medicine. [ABSTRACT FROM AUTHOR]

Background: The majority of patients with lung cancer exhibit drug resistance after anti-PD-1 immunotherapy, leading to shortened patient survival time. Previous studies have suggested an association between epigenetic abnormalities such as methylation and clinical response to anti-PD-1 immunotherapy, while the role of SUMOylation in resistance to anti-PD-1 antibody immunotherapy is still unclear. Methods: Here, the mRNA expression of 15 SUMO machinery components in PBMC from lung cancer patients receiving anti-PD-1 immunotherapy were analyzed using real-time PCR. Base on the percentage change in mRNA levels, the relationship between the expression of SUMO machinery components and outcomes of anti-PD-1 immunotherapy, and the influencing factors of SUMOylation were evaluated. PBMC was treated with different concentrations of 2-D08 (a specific inhibitor of SUMOylation) in vitro, and analyzed the activation and the death rates of lymphocyte subsets by flow cytometry analysis. Results: A predictive method, base on the gene expression of three SUMO machinery components (SUMO1, SUMO3 and UBE2I), were developed to distinguish non-responders to PD-1 inhibitors. Furthermore, the number of lymphocytes in peripheral blood significantly reduced in the dysregulated SUMOylation groups (the percentage change >100 or -50 ~ -100 groups). In vitro studies confirmed that lightly low SUMOylation level improved the activation status of T and NK lymphocytes, but extremely low SUMOylation level lead to the increased death rates of lymphocytes. Conclusion: Our findings implied that dysregulated gene expression of SUMO machinery components could induce the resistance of anti-PD-1 immunotherapy in lung cancer by upregulating the death of peripheral blood lymphocytes. These data might provide effective circulating biomarkers for predicting the efficacy of anti-PD-1 immunotherapy, and uncovered a novel regulatory mechanism of resistance to anti-PD-1 immunotherapy. [ABSTRACT FROM AUTHOR]

The use of flow cytometry in mice is constrained by several factors, including the limited availability of mouse-specific antibodies and the need to work with small volumes of peripheral blood. This is particularly challenging for longitudinal studies, as serial blood samples should not exceed 10% of the total blood volume in mice. To address this, we have developed two novel flow cytometry panels designed to extensively analyze immune cell populations in mice during longitudinal studies, using only 50 µL of peripheral blood per panel. Additionally, a third panel has been designed to conduct a more detailed analysis of cytotoxic and inhibitory markers at the end point. These panels have been validated on a lipopolysaccharide (LPS)-induced lung inflammation model. Two experiments were conducted to 1) validate the panels' sensitivity to immune challenges (n=12) and 2) to assess intrinsic variability of measurements (n=5). In both experiments, we collected 50 µL of peripheral blood for each cytometry panel from the maxillary venous sinus. All antibodies were titrated to identify the optimal concentration that maximized the signal from the positive population while minimizing the signal from the negative population. Samples were processed within 1 hour of collection using a MACSQuant Analyzer 16 cytometer. Our results demonstrate that these immunological panels are sensitive enough to detect changes in peripheral blood after LPS induction. Moreover, our findings help determine the sample size needed based on the immune population variability. In conclusion, the panels we have designed enable a comprehensive analysis of the murine immune system with a low blood volume requirement, enabling the measure of both absolute values and relative percentages effectively. This approach provides a robust platform for longitudinal studies in mice and can be used to uncover significant insights into immune responses. [ABSTRACT FROM AUTHOR]