Correlation analysis between peripheral blood dendritic cell subsets and PD-1 in patients with peritoneal adenocarcinoma

Abstract The aim of this study was to explore the association between differential percentages of dendritic cell (DC) subsets in peripheral blood and malignancy (grade and lymph node metastasis) of peritoneal adenocarcinoma patients and the frequencies of dendritic cell subsets in the normal controls. The peripheral blood of 30 patients with peritoneal adenocarcinoma and 12 healthy controls were collected for multicolor flow cytometry analysis. Peritoneal adenocarcinoma patients were grouped according to the malignant degree (grade and lymph node metastasis). Percentages of myeloid DCs (mDCs) and its subsets MDC1 and MDC2 in DCs were lower in peripheral blood of patients with peritoneal adenocarcinoma than in normal controls. The percentages of plasmacytoid dendritic cells (pDCs) and CD16+mDCs in DCs were higher than in normal controls. Compared with poor differentiation grade, patients with well/moderate differentiation grade had an increased percentage of CD16+mDCs. Contrary to CD16+mDCs, the percentage of MDC1 was lower in the well/moderate differentiation grade group. In patients with no lymph node metastasis, pDCs and CD16+mDCs levels were higher compared with patients with lymph node metastasis. mDCs and MDC1 levels had opposite results. pDCs were positively correlated with CD16+mDCs in peripheral blood of peritoneal patients, as was mDCs and MDC1. CD16+mDCs were negatively correlated with MDC1. The percentages of pDCs and CD16+mDCs in DCs were positively correlated with CD3+CD8+T cells, and pDCs also positively correlated with CD8+PD-1+T cells. Our results revealed that DCs subsets correlated with peritoneal adenocarcinoma malignancy. Dendritic cells play an independent role in the immune function of peritoneal adenocarcinoma.

Alpha-synuclein dynamics bridge Type-I Interferon response and SARS-CoV-2 replication in peripheral cells

BACKGROUND: Increasing evidence suggests a double-faceted role of alpha-synuclein (α-syn) following infection by a variety of viruses, including SARS-CoV-2. Although α-syn accumulation is known to contribute to cell toxicity and the development and/or exacerbation of neuropathological manifestations, it is also a key to sustaining anti-viral innate immunity. Consistently with α-syn aggregation as a hallmark of Parkinson's disease, most studies investigating the biological function of α-syn focused on neural cells, while reports on the role of α-syn in periphery are limited, especially in SARS-CoV-2 infection. RESULTS: Results herein obtained by real time qPCR, immunofluorescence and western blot indicate that α-syn upregulation in peripheral cells occurs as a Type-I Interferon (IFN)-related response against SARS-CoV-2 infection. Noteworthy, this effect mostly involves α-syn multimers, and the dynamic α-syn multimer:monomer ratio. Administration of excess α-syn monomers promoted SARS-CoV-2 replication along with downregulation of IFN-Stimulated Genes (ISGs) in epithelial lung cells, which was associated with reduced α-syn multimers and α-syn multimer:monomer ratio. These effects were prevented by combined administration of IFN-ß, which hindered virus replication and upregulated ISGs, meanwhile increasing both α-syn multimers and α-syn multimer:monomer ratio in the absence of cell toxicity. Finally, in endothelial cells displaying abortive SARS-CoV-2 replication, α-syn multimers, and multimer:monomer ratio were not reduced following exposure to the virus and exogenous α-syn, suggesting that only productive viral infection impairs α-syn multimerization and multimer:monomer equilibrium. CONCLUSIONS: Our study provides novel insights into the biology of α-syn, showing that its dynamic conformations are implicated in the innate immune response against SARS-CoV-2 infection in peripheral cells. In particular, our results suggest that promotion of non-toxic α-syn multimers likely occurs as a Type-I IFN-related biological response which partakes in the suppression of viral replication. Further studies are needed to replicate our findings in neuronal cells as well as animal models, and to ascertain the nature of such α-syn conformations.

Establishment of a 21-color Panel for the Detection of Immune Cell Subsets in Human Non-small Cell Lung Cancer Tumor Tissues with Flow Cytometry / 中国肺癌杂志

BACKGROUND@#With the rise of multicolor flow cytometry, flow cytometry has become an important means to detect the immune microenvironment of lung cancer, but most of them are used to detect the proportion of cell subsets or the function of major cell subsets, and they cannot be detected at the same time. Therefore, a reliable 21-color flow cytometry protocol was established to detect the immune cell subsets in human non-small cell lung cancer (NSCLC) tumor tissues.@*METHODS@#Cell membrane surface antibodies cluster of differentiation (CD)45, CD3, CD19, CD4, CD8, programmed cell death 1 (PD-1), CD39, CD103, CD25, CD127, chemokine receptor 8 (CCR8), CD56, CD11c, human leukocyte antigen (HLA)-DR, CD38, CD27, CD69, CD62L, CD45RA, CCR7 and nucleic acid dye L/D were used to develop the protocol. Firstly, antibody titration experiments, voltage optimization, subtraction of one color staining and single color staining experiments were carried out for each antibody, and after the experimental conditions and detection schemes were determined, the feasibility of the scheme was verified by using peripheral blood mononuclear cells (PBMCs) specimens of six healthy adult volunteers. Tumor tissue samples from 6 NSCLC patients were tested and analyzed.@*RESULTS@#The established 21-color flow cytometry protocol was used to detect the tumor tissue samples of 6 NSCLC patients, and the proportion of each cell subset in lung cancer tissue, as well as the immunophenotype and differentiation of the main cell population, were analyzed.@*CONCLUSIONS@#The successfully established 21-color flow cytometry protocol is suitable for the detection of PBMCs and NSCLC tissue samples, which provides an effective new idea for monitoring the immune microenvironment status in lung cancer.

Natural killer cells 56bnght16- have higher counts in the umbilical cord blood than in the adult peripheral blood

ABSTRACT Introduction and hypothesis: Umbilical cord blood (UCB) is an alternative source of hematopoietic stem cells for allogeneic hematopoietic stem cell transplantation in the absence of a compatible donor. The UCB transplantation has a lower incidence of chronic graft versus host disease (GvHD), but is associated with slower engraftment and slower immune reconstitution, compared to other sources. Dendritic cells (DCs) and Natural Killer cells (NKs) play a central role in the development of GvHD and the graft versus leukemia (GvL) effect, as well as in the control of infectious complications. Method: We quantified by multiparametric flow cytometry monocytes, lymphocytes, NK cells, and DCs, including their subsets, in UCB samples from 54 healthy newborns and peripheral blood (PB) from 25 healthy adult volunteers. Results: In the UCB samples, there were higher counts of NK cells 56bright16- (median 0.024 × 109/L), compared to the PB samples (0.012 × 109/L, p < 0.0001), NK 56dim16bright (median 0.446 × 109/L vs. 0.259 × 109/L for PB samples, p = 0.001) and plasmacytoid dendritic cells (pDCs, median 0.008 × 109/L for UCB samples vs. 0.006 × 109/L for PB samples, p = 0.03). Moreover, non-classic monocyte counts were lower in UCB than in PB (median 0.024 × 109/L vs. 0.051 × 109/L, respectively, p < 0.0001). Conclusion: In conclusion, there were higher counts of NK cells and pDCs and lower counts of non-classic monocytes in UCB than in PB from healthy individuals. These findings might explain the lower incidence and severity of chronic GvHD, although maintaining the GvL effect, in UCB transplant recipients, compared to other stem cell sources.

COMPARATIVE STUDY OF AUTOMATED CELL COUNTER HISTOGRAM AND PERIPHERAL BLOOD SMEAR IN DIAGNOSIS OF ANEMIA

Introduction: Over the past few years, complete blood count, RBC histogram and peripheral blood smear have become the important diagnostic tools to diagnose various haematological conditions. Major public health burden worldwide is anemia with high prevalence in developing countries like India1. Red blood cell and histogram are indispensable for diagnosis and management of anemia2. The major diagnostic tool for work up of most commercial laboratories has been analysing the blood film routinely. The aim AIM: of the study is to compare between automated cell counter histogram and peripheral smear finding in diagnosis of anemia. Material and method: A prospective comparative study of RBC histogram and peripheral blood smear in diagnosis of anemia was done on 100 patients of HB<14gm%, over six month time span (June2022-Nov2022) in the central laboratory of Saraswathi Institute of Medical Sciences Hapur, (UP) India. This study included all the age groups. All cases of anemia that have undergone blood transfusion is excluded from this study. In Result: our study it was observed that on peripheral blood smear, the most common type of anemia was microcytic hypochromic anemia followed by normocytic normochromic anemia, when we compared with automated cell counter generated histogram most common type of anemia was normocytic normochromic followed by microcytic hypochromic anemia. In our study female population were more than males. The mean age was(32.4yr). Automated cell counters generated CBC histograms and peripheral blood smears Conclusion: plays a major role in diagnosis and management of red cell disorder. Our study observed that histogram patterns and their confirmation by peripheral blood smear along with clinical history gives accurate and confirmed diagnosis of various haematogical conditions. There is much improvement in accuracy and precision 3 reducing the subjective errors .

Alteraciones hematológicas en sangre periférica de pacientes con COVID-19 / Molecular study of leukemias: essential tool for the hematologist

Introducción: Los primeros casos con neumonía atípica de etiología desconocida fueron reportados en Wuhan, China en diciembre de 2019. En enero 2020 se describió como agente causal un nuevo tipo de virus de la familia Coronaviridae, denominado SARS-CoV-2. Objetivo: Evaluar la significación clínica de los cambios hematológicos y morfológicos en la sangre periférica de pacientes con COVID-19. Métodos: Se realizó un estudio descriptivo, observacional, transversal que incluyó a los pacientes con COVID-19 que ingresaron en el Hospital Clínico Quirúrgico Docente Freyre de Andrade desde el 1ro de junio hasta 31 de septiembre de 2021. Los pacientes fueron asignados a dos grupos según fueron admitidos, en las unidades de vigilancia intensiva o en la unidad de cuidados intensivos. Se les realizó hemograma completo y lámina periférica el día del ingreso para evaluar la significación clínica de estas variables en la evolución de estos pacientes. Resultados: El sexo femenino predominó en los pacientes ingresados en unidades de vigilancia intensiva (67,36 %) y el masculino en los ingresados en unidades de cuidados intensivos (63,26 %). La media de edad fue mayor en el grupo de pacientes en cuidados intensivos (65,83 años). La leucocitosis y el menor recuento de plaquetas predominaron en los pacientes ingresados en cuidados intensivos, seguido de linfopenia. Las macroplaquetas, las vacuolas citoplasmáticas y las granulaciones tóxicas fueron más frecuentes en el grupo de cuidados intensivos. Conclusiones: El hemograma y el frotis de sangre periférica son útiles para diagnosticar y predecir la evolución de los pacientes y permiten un mejor manejo de la infección.

Introduction: The first cases of atypical pneumonia of unknown etiology were reported in Wuhan, China in December 2019. In January 2020 a new virus from Coronaviridae family was described as causal agent and was named SARS-COV-2. Objectives: To evaluate the clinical significance of numerical values of complete blood count (CBC) and morphologic changes on peripheral blood on patients with COVID-19. Methods: A descriptive, observational, transversal study included patients with diagnosis of COVID-19 admitted in Freyre de Andrade Hospital in Havana, between June 1st and September 31st of 2022 was carried out. Patients were assigned to two groups according to their admission in intensive vigilance ward or intensive care unit. CBC test and peripheral blood smear were performed on admission day to evaluate the significance on clinical evolution. Results: Female sex predominated on intensive vigilance group (67,36 %) and male in intensive care group (63,26 %). Media of age was superior in intensive care group (67,83 years). Leukocytosis and low level of platelets count were significantly more common in more severe group followed by lymphopenia. The presence of big platelets, cytoplasmic vacuoles and toxic granules were more common in intensive care unit group. Conclusions: The CBC and peripheral blood smear are useful tools to diagnose and predict clinical evolution and allow a better management of infection.

Calcitonin gene-related peptide (CGRP) concentrations outside migraine attacks in peripheral blood as a potential biomarker for chronic migraine / Concentrações de peptídeo relacionado ao gene da calcitonina (CGRP) fora dos ataques de enxaqueca no sangue periférico como um potencial biomarcador para enxaqueca crônica

Introduction: Chronic migraine is a debilitating condition that affects a significant portion of the population. Accurate diagnosis and treatment of chronic migraine remain a challenge due to the lack of objective biomarkers. Calcitonin gene-related peptide (CGRP) is a neuropeptide involved in the pathophysiology of migraine and has been proposed as a potential biomarker for migraine. Methods: We measured CGRP levels in peripheral blood samples collected from 142 participants with chronic or episodic migraine and 24 healthy controls during ictal periods, i.e., outside migraine attacks. We compared CGRP levels between the three groups and assessed the correlation between CGRP levels and clinical features of chronic migraine. Conclusion: Our study provides evidence that CGRP levels in peripheral blood during ictal periods may serve as a potential biomarker for chronic migraine. Further studies are needed to validate these findings and to explore the clinical utility of CGRP as a biomarker for chronic migraine.

Introdução: A enxaqueca crônica é uma condição debilitante que afeta uma parcela significativa da população. O diagnóstico preciso e o tratamento da enxaqueca crónica continuam a ser um desafio devido à falta de biomarcadores objetivos. O peptídeo relacionado ao gene da calcitonina (CGRP) é um neuropeptídeo envolvido na fisiopatologia da enxaqueca e foi proposto como um potencial biomarcador para enxaqueca. Métodos: Medimos os níveis de CGRP em amostras de sangue periférico coletadas de 142 participantes com enxaqueca crônica ou episódica e 24 controles saudáveis ​​durante períodos ictais, ou seja, fora das crises de enxaqueca. Comparamos os níveis de CGRP entre os três grupos e avaliamos a correlação entre os níveis de CGRP e as características clínicas da enxaqueca crônica. Conclusão: Nosso estudo fornece evidências de que os níveis de CGRP no sangue periférico durante os períodos ictais podem servir como um potencial biomarcador para enxaqueca crônica. Mais estudos são necessários para validar estes resultados e explorar a utilidade clínica do CGRP como biomarcador para enxaqueca crónica.

Correlation Between Peripheral Blood Biomarkers and Immune Checkpoint Inhibitors in Treatment of Advanced Esophageal Cancer / 肿瘤防治研究

Objective To explore the effect of peripheral blood markers on the efficacy and prognosis of patients with advanced esophageal cancer treated with immune checkpoint inhibitors (ICIs). Methods The case data of 61 patients with advanced esophageal cancer who met the inclusion criteria were collected. Data on clinical indicators and peripheral blood markers as well as objective response rate (ORR) and progression-free-survival (PFS) were obtained. Results The median PFS of the included patients was 7.10 months (95%CI: 5.12-9.07). The ORR of patients with baseline lactate dehydrogenase (LDH) < 201 was better than that of patients with LDH≥201 (P < 0.05). Univariate analysis showed that baseline LDH0 < 201, neutrophil to lymphocyte ratio (NLR) < 3.9, platelet-to-lymphocyte ratio (PLR) < 240.3, and LDH1 < 249.0 two weeks after ICI treatment were significantly associated with significant improvement in PFS (P < 0.05). In multivariate analysis, patients with NLR0 < 3.9 had longer PFS (P < 0.05). Conclusion LDH0 < 201, NLR0 < 3.9, PLR0 < 240.3, and LDH1 < 249.0 are positively correlated with the prognosis of patients with advanced esophageal cancer treated with ICIs.

Role of Preoperative Peripheral Blood Inflammatory Parameters and Postoperative Lymph-node Ratio in Prognosis of Patients with Gastric Cancer Undergoing Chemotherapy / 肿瘤防治研究

Objective To analyze the role of preoperative peripheral blood inflammatory parameters and postoperative lymph-node ratio (LNR) in the prognosis of gastric cancer patients treated with chemotherapy. Methods The neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), systemic immune inflammatory response index (SII), lymphocyte-to-monocyte ratio (LMR), Onodera prognostic nutritional index (OPNI), and LNR of 108 patients with gastric cancer were classified into high and low groups to analyze their prognostic value on the overall survival (OS) of gastric cancer patients treated with chemotherapy. The independent prognostic indicators were plotted in columns to predict the survival rate of gastric cancer patients. Results NLR was statistically significant in the prognostic assessment of gastric cancer patients treated with chemotherapy (P < 0.001). Moreover, the high PLR group, high SII group, high LNR group, N3 stage, TNM (Ⅲ-Ⅳ) stage, nerve invasion, and carcinoembryonic antigen (CEA) were independent risk factors for the prognosis of gastric cancer (all P < 0.05). These findings indicated that in predicting the prognosis of gastric cancer, the combination of LNR, PLR, and SII (AUC=0.875) was better than that of LNR, PLR, and SII alone or in pairwise combination. Conclusion Elevated NLR, LNR, PLR, and SII are associated with significantly reduced survival of patients. LNR, PLR, and SII could be used as independent risk factors for the prognosis of gastric cancer patients treated with chemotherapy, and their combination can predict the survival of gastric cancer patients.

Peripheral Blood Laboratory Test Results Combined with TCF1+CD8+ T Lymphocytes Ratio to Predict the Response and Prognosis of Immunotherapy to Advanced Lung Cancer / 中国肺癌杂志

BACKGROUND@#Immune checkpoint inhibitors (ICIs) therapy lacks viable biomarkers for response and prognosis prediction. This study aimed to investigate the correlation of peripheral blood laboratory test results combined with lymphocyte subset ratios to the response and prognosis of immunotherapy in advanced lung cancer.@*METHODS@#Advanced lung cancer patients admitted to West China Hospital, Sichuan University from May 2021 to July 2023 were prospectively enrolled in this study. Clinical data and peripheral blood were collected before and after treatment and lymphocyte subset ratios were analyzed by flow cytometry. Logistic regression was used to identify factors correlated to ICIs treatment efficacy. Cox modeling was applied to explore the prognostic factors.@*RESULTS@#Logistic regression showed that the baseline level of transcription factor T cell factor 1 (TCF1)+CD8+ T cell ratio and peripheral white blood cell (WBC) count, lymphocyte percentage, cytokeratin 19 fragment (CYFRA21-1) after 1 cycle of ICIs treatment were the potential predictors for ICIs response (P<0.05). Cox regression analysis showed that the baseline level of TCF1+CD8+ T cell ratio (P=0.020) and peripheral WBC count after 1 cycle of ICIs treatment (P<0.001) were prognostic factors.@*CONCLUSIONS@#Patients with high baseline TCF1+CD8+ T cell ratio combined with low WBC counts and low CYFRA21-1 level after 1 cycle of ICIs treatment are more likely to benefit from ICIs therapy.

Clinical Analysis of Autologous Peripheral Blood Hematopoietic Stem Cells Mobilization and Collection in Lymphoma Patients / 中国实验血液学杂志

OBJECTIVE@#To analyze the factors influencing collection of autologous peripheral blood hematopoietic stem cells in lymphoma patients.@*METHODS@#Clinical data of 74 patients who received autologous peripheral blood hematopoietic stem cells mobilization and collection in the 940th Hospital of Joint Logistic Support Force of PLA from April 2009 to April 2021 were collected. The effects of gender, age, disease type, stage, course of disease, chemotherapy cycle number, relapse, radiotherapy, disease status and blood routine indexes on the day of collection on peripheral blood hematopoietic stem cell collection were analyzed.@*RESULTS@#The success rate of collection was 95.9%(71/74), and the excellent rate of collection was 71.6%(53/74). There was a significantly statistical differentce in the number of CD34+ cells in grafts collected from patients with chemotherapy cycle ≤6 and >6 ［(9.1±5.2)×106/kg vs (6.4±3.7)×106/kg, P=0.031］. The number of CD34+ cells in the first collection was positively correlated with WBC count, hemoglobin, platelet count, neutrophil count, lymphocyte count, monocyte count and hematocrit value on the day of collection ( r value was 0.424,0.486,0.306,0.289,0.353,0.428,0.528, respectively). WBC count, hemoglobin, monocyte count and hematocrit value have higher predictive value for the first collection of CD34+ cells. The area under the receiver operating characteristic was 0.7061,0.7845,0.7319,0.7848, respectively.@*CONCLUSION@#Low dose CTX and VP16 chemotherapy combined with G-CSF can effectively mobilize autologous peripheral blood stem cells. The cycle number of chemotherapy relates to the collection of autologous peripheral blood stem cells. After mobilization, the success of the first collection can be better predicted by the blood routine indexes.

Efficacy and Safety of Etoposide Combined with Cyclophosphamide for Autologous Peripheral Blood Stem Cell Mobilization in Patients with Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To evaluate the efficacy and safety of etoposide combined with cyclophosphamide (EC) regimen for mobilization of autologous peripheral blood stem cells (APBSCs) in patients with multiple myeloma (MM).@*METHODS@#The clinical data of 48 MM patients who received APBSC transplantation (APBSCT) in Department of Hematology of the First Affiliated Hospital of Nanchang University from January 2015 to October 2021 were retrospectively analyzed. The mobilization success rate and mobilization optimal rate of EC regimen were counted, and its effect on transplant efficacy, adverse reactions, hematopoietic reconstitution after transplantation, and survival time of MM patients were analyzed.@*RESULTS@#APBSCs were collected on day 14 (10-19) after EC administration. The median of collected CD34+ cells was 6.82 (1.27-22.57)×106/kg, and the median number of apheresis session was 2 (1-4). The mobilization success rate (collecting CD34+ cells≥2×106 cells/kg after completion of apheresis) was 98% (47/48), and mobilization optimal rate (collecting CD34+ cells≥5×106 cells/kg after completion of apheresis) was 71% (34/48). The depth of remission were improved after APBSCT, and the complete remission (CR) rate increased from 45.8% before transplantation to 87.5% after transplantation (P <0.01). There was no transplant-related death, no blood transfusion during mobilization, and no mucositis occurred in the patients. The most common complication was neutropenia, with an incidence of 75.0% (36/48). After transplantation, all the patients successfully achieved hematopoietic reconstitution. The median time to neutrophil engraftment was 10 (9-26) days, and median time to platelet engraftment was 10 (8-33) days. By the end of follow-up, both the median progression-free survival (PFS) and overall survival (OS) time were not reached. The 5-year estimated PFS rate and OS rate was 53.8% and 82.4%, respectively.@*CONCLUSION@#The EC regimen for mobilization of APBSC has a high acquisition success rate and controllable adverse reactions, which can be an effective and safe mobilization regimen in MM patients.

Comparison of Plerixafor or Cyclophosphamide Combined with G-CSF in Mobilization of Peripheral Blood Stem Cells in Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To compare the efficacy of plerixafor (PXF) combined with granulocyte colony-stimulating factor (G-CSF) (PXF+G-CSF) and cyclophosphamide (Cy) combined with G-CSF (Cy+G-CSF) in the mobilization of peripheral blood stem cells (PBSCs) in patients with multiple myeloma (MM).@*METHODS@#The clinical data of 41 MM patients who underwent PBSC mobilization using PXF+G-CSF (18 cases) or Cy+G-CSF (23 cases) in Shanxi Bethune Hospital from January 2019 to December 2021 were retrospectively analyzed, including the count of collected CD34+ cells, acquisition success rate, failure rate, and optimal rate. The correlation of sex, age, disease type, DS staging, ISS staging, number of chemotherapy cycle, disease status before mobilization, and mobilization regimen with the collection results was analyzed, and the adverse reactions, length of hospital stay, and hospitalization costs were compared between the two mobilization regimens.@*RESULTS@#The 41 patients underwent 97 mobilization collections, and the median number of CD34+ cells collected was 6.09 (0-34.07)×106/kg. The acquisition success rate, optimal rate, and failure rate was 90.2%, 56.1%, and 9.8%, respectively. Univariate analysis showed that sex, age, disease type, and disease stage had no significant correlation with the number of CD34+ cells collected and acquisition success rate (P >0.05), but the patients with better disease remission than partial remission before mobilization were more likely to obtain higher CD34+ cell count (P <0.05). The PXF+G-CSF group had a larger number of CD34+ cells and higher acquisition success rate in the first collection than Cy+G-CSF group (both P <0.05), and had lower infection risk and shorter length of hospital stay during mobilization (both P <0.05), but the economic burden increased (P <0.05).@*CONCLUSION@#PXF+G-CSF used for PBSC mobilization in MM patients has high first acquisition success rate, large number of CD34+ cells, less number of collection times, and short length of hospital stay, but the economic cost is heavy.

Efficacy and Safety of Plerixafor Combined with G-CSF for Autologous Peripheral Blood Hematopoietic Stem Cell Mobilization in Lymphoma Patients / 中国实验血液学杂志

OBJECTIVE@#To investigate the efficacy and safety of plerixafor combined with granulocyte colony-stimulating factor (G-CSF) in mobilizing peripheral blood hematopoietic stem cells in patients with lymphoma.@*METHODS@#The clinical data of lymphoma patients who received autologous hematopoietic stem cell mobilization using plerixafor combined with G-CSF from January 2019 to December 2021 were retrospectively analyzed. The patients received 3 kinds of mobilization regimens: front-line steady-state mobilization, preemptive intervention, and recuse mobilization. The acquisition success rate, excellent rate of collection, and incidence of treatment-related adverse reaction were counted. The influence of sex, age, disease remission status, bone marrow involvement at diagnosis, chemotherapy lines, number of chemotherapy, platelet count and number of CD34+ cells on the day before acquisition in peripheral blood on the collection results were analyzed to identify the risk factors associated with poor stem cell collection.@*RESULTS@#A total of 43 patients with lymphoma were enrolled, including 7 cases who received front-line steady-state mobilization, 19 cases who received preemptive intervention, and 17 cases who received recuse mobilization. The overall acquisition success rate was 58.1% (25/43) after use of plerixafor combined with G-CSF, and acquisition success rate of front-line steady-state mobilization, preemptive intervention, and recuse mobilization was 100%, 57.9%(11/19), and 41.2%(7/17), respectively. The excellent rate of collection was 18.6%(8/43). A total of 15 patients experienced mild to moderate treatment-related adverse reactions. The number of CD34+ cells < 5 cells/μl in peripheral blood on the day before collection was an independent risk factor affecting stem cell collection.@*CONCLUSIONS@#Plerixafor combined with G-CSF is a safe and effective mobilization regimen for patients with lymphoma. The number of CD34+ cells in peripheral blood on the day before collection is an predictable index for the evaluation of stem cell collection.

Research update on DNA methylation in peripheral blood cells as a risk factor for coronary heart disease / 中华检验医学杂志

Coronary heart disease (CHD) is a kind of cardiovascular diseases originated from atherosclerosis (AS), and chronic inflammation is one of the pathological characteristics. The peripheral blood leukocytes, especially mononuclear cells, play an important role in the AS processes. Recently, in a series of Epigenome-Wide Association Studies (EWAS), multiple DNA differential methylation sites in peripheral blood cells were found to be statistically associated with CHD, which suggested that these DNA differential methylation sites might serve as new risk factors for CHD. The recognition of the variant of DNA methylation as a common epigenetic nucleic acid modification in the occurrence and development of CHD, is ongoing. DNA methylation has the potential to become warning biomarkers, which might provide new ideas and evidences for mechanistic studies of CHD.

Expression of progranulin in mononuclear cells in the peripheral blood of patients with post-stroke aphasia / 中华物理医学与康复杂志

Objective:To document the expression of aphasia-related progranulin gene (GRN) in mononuclear cells in the peripheral blood (PBMC) of patients with post-stroke aphasia (PSA).Methods:PC12 cells at the logarithmic-growth stage were cultured and divided into a non-specific interference group (the gene control group) and a specific interference group (the gene silencing group) when the cell density reached 30 to 50%. After the expression of GRN was knocked down in the cells, the occurrence of variable splicing events was analyzed using high-throughput transcriptome sequencing (RNA-seq). Meanwhile, 10 PSA patients were selected into a patient group and 10 healthy counterparts were chosen as a control group. Blood was collected from both groups and real-time fluorescence quantitative polymerase chain reactions (RT-qPCR) were employed to determine any changes in GRN mRNA expression and the occurrence of variable splicing events in the nuclear factor related to kappa-B-binding protein (NFRKB) in their PBMCs. The patient group received conventional speech therapy, and immediately after their first and second blood collections their speech functioning was assessed using the Chinese Aphasia Battery (ABC). Pearson correlation coefficients were then computed relating the GRN expression and ABC scores.Results:After knocking down GRN in the PC12 cells, the expression of GRN in the gene knockdown group was significantly different from that in the control group. There were 237 genes with significant differences in variable splicing between the two samples. The number of genes with variable splicing events at the 5′ end was the largest. There were also significant differences between the groups in the average occurrence of NFRKB variable splicing events. And significant diffe-rences were observed in the mRNA expression of GRN between the two blood collections from the patient group, as well as between the first collection from the patient group and the controls. The average oral expression score of the PSA patients improved significantly, particularly the retelling score. The changes in the GRN expression level were positively correlated with the recovery of oral expression ability.Conclusion:GRN can promote the recovery of speech function in PSA patients by regulating the variable splicing of NFRKB.

Correlations of transglutaminase 2 expression in peripheral blood mononuclear cells with relevant inflammatory factors and disease severity in patients with atopic dermatitis / 中华皮肤科杂志

Objective:To determine the expression of transglutaminase 2 (TGM2) in peripheral blood mononuclear cells (PBMCs) from patients with atopic dermatitis (AD), and to analyze its correlation with AD-related inflammatory factors and disease severity.Methods:A total of 29 AD patients and 15 healthy controls were collected from the First Affiliated Hospital of Fujian Medical University from July 2020 to January 2021. Ten milliliters of peripheral blood samples were collected from each subject, so was the clinical information, including age, gender, course of disease, eosinophil counts, basophil counts, total IgE levels, Scoring AD index (SCORAD), etc. PBMCs were isolated by density gradient centrifugation. Fluorescence-based quantitative PCR was performed to determine the mRNA expression of TGM2 and AD-related inflammatory factors (interleukin [IL]-1β, IL-4, IL-6, IL-8, IL-10, IL-13, IL-17, thymic stromal lymphopoietin [TSLP], P2RX7 [purinergic receptor P2X, ligand-gated ion channel, 7], etc.) in PBMCs from 29 AD patients and 15 healthy controls, and flow cytometry to determine TGM2 protein expression on PBMCs. Mann-Whitney U test was used to analyze differences between groups, and Spearman correlation analysis to evaluate the correlation. Results:The relative mRNA expression of TGM2 in PBMCs did not differ between the AD group and control group ( M[ Q1, Q3]: 0.509 [0.325, 0.958] vs. 0.475 [0.328, 1.051], U = 210.50, P = 0.872). Compared with the control group, the AD group showed significantly decreased IL-4 mRNA expression (0.171[0.049, 0.449] vs. 0.824 [0.397, 1.378], P < 0.001), but significantly increased mRNA expression of IL-8 and IL-13 ( P = 0.011, 0.006, respectively). Spearman correlation analysis showed that the mRNA expression level of TGM2 in PBMCs was positively correlated with the mRNA expression levels of IL-4 and P2RX7 in the AD group ( rs = 0.42, 0.40, P = 0.024, 0.034, respectively), while there were no correlations between TGM2 mRNA expression and AD severity-related indicators (all P>0.05), such as age (21[16, 29] years), course of disease (4[1,10] years), eosinophil counts (0.33[0.18, 0.65] × 10 9/L), basophil counts (0.04[0.03, 0.06] × 10 9/L], SCORAD scores (60.5[46.98, 66.13] points), and serum total IgE levels (373 [40, 1 815] IU/ml). The relative protein expression levels of TGM2 on the surface of PBMCs did not differ between the AD group and control group (54.9 [47.6, 62.8] vs. 55.55 [51.5, 60.25], U = 112.00, P = 0.922) ], and no correlations were observed between the protein expression of TGM2 on PBMCs and AD severity-related indicators in the AD group (all P > 0.05) . Conclusion:No significant differences were observed in TGM2 mRNA expression in PBMCs or TGM2 protein expression on the surface of PBMCs between the AD patients and healthy controls, and there were no correlations between the TGM2 mRNA and protein expression and AD severity.

Biomarkers related to the treatment of melanoma with immune checkpoint inhibitors / 中华皮肤科杂志

Recently, great breakthroughs have been made in the treatment of melanoma with immune checkpoint inhibitors. However, only a small proportion of patients show a long-lasting response to immunotherapy, and risks of immune-related adverse events and drug resistance have been also increasing along with the emergence of combination treatment. This review summarizes biomarkers related to the efficacy of immune checkpoint inhibitors in the treatment of melanoma, aiming to predict and screen out patients who may benefit from immunotherapy, guide individualized clinical treatment, and reduce the occurrence of drug resistance and adverse reactions.

Difference and its clinical significance of peripheral blood lymphocyte subsets between elderly and non-elderly patients with rheumatoid arthritis / 中华老年医学杂志

Objective:To investigate the difference of lymphocyte subsets between elderly patients with rheumatoid arthritis and non-elderly patients and its clinical significance.Methods:A total of 124 patients with rheumatoid arthritis in Affiliated Hospital of Nantong University from January, 2017 to December, 2019 were enrolled.The patients were divided into elderly group(≥60 years old, 34 cases)and non-elderly group(<60 years old, 90 cases). Rheumatoid arthritis activity(DAS-28)scoring was performed for each patient.Peripheral blood mononuclear cells(PBMCs)were extracted by Ficoll density centrifugation.Lymphocytes were labeled and detected by 18-color flowcytometry with more than 30 fluorescent antibodies.Results:DAS-28 scoring showed that the disease activity score of the elderly group(4.56±1.89)was higher than that of the non-elderly group(3.37±1.49)( t=3.633, P<0.001). Flow cytometry showed that MAIL%T(mucus-associated lymphoid tissue T cell subset)( Z=-2.798, P=0.005), Tn%CD8 T cells(initial CD8 T cells)( Z=-2.179, P=0.029), VD2% T(Vδ2+ T, γδT cell subtype)( Z=-2.806, P=0.005), PD1-CD28-%Th( Z=-2.050, P=0.040)and IGM+ D-%B( Z=-2.376, P=0.017)were lower in the elderly group than in the non-elderly group.While, CD45+ CD27+ %CD8 T cells( Z=-3.069, P=0.002), abT%T cell(αβT cells)( Z=-2.103, P=0.035), CD27-CD28+ %T cells( Z=-2.341, P=0.019), ASC%PBMC( Z=-2.341, P=0.019)and ASC%CD19+ ( Z=-2.000, P=0.046)subgroup expression were higher in the elderly group than in the non-elderly group. Conclusions:The disease activity of elderly patients with rheumatoid arthritis is significantly higher than that of younger patients.The expressions of abT%T and CD4% abT in effector T cells of elderly patients with rheumatoid arthritis are higher than those of younger patients, while the expression of VD2% T is lower.The expression level of CD45RA+ CD27+ %CD8 T with cytotoxic effect is higher; However, the expression level of Tn%CD8 T in naive cells is lower.

Analysis of the efficiency and influencing factors of peripheral blood hematopoietic stem cell collection / 中国医师进修杂志

Objective:To investigate the collection efficiency of peripheral blood hematopoietic stem cells and explore its influencing factors.Method:The parameters of the collection process, blood routine indexes and the number of MNC and CD 34+ cells of the product were detected by Fresenius blood cell separator, Mindray blood cell analyzer and BD flow cytometer. A retrospective analysis was performed on 72 patients who underwent autologous peripheral blood hematopoietic stem cell transplantation in Southwest Hospital of Army Medical University from January 2013 to January 2021, including the correlation among gender, age, blood routine indexes, collection circulation volume and MNC and CD 34+ cell count in these cases, and influence of various factors on collection efficiency of peripheral blood stem cells. The correlation among gender, age, blood routine indexes, collection circulation volume and MNC and CD 34+ cell count in 72 cases of autologous transplantation patients, and influence of various factors on collection efficiency of peripheral blood stem cells were analyzed retrospectively. Results:There were no significant differences in collecting efficiency of peripheral blood stem cells among patients with different age, sex and disease type ( P>0.05). The collected MNC count of all patients was positively correlated with the collection cycle count ( r = 0.33, P<0.001) and WBC count after mobilization ( r = 0.41, P<0.001). The number of CD 34+ cells collected was positively correlated with MNC count after mobilization ( r = 0.38, P<0.001) and the amount of white membrane collected ( r = 0.48, P<0.001). Logistic multivariate regression analysis showed that MNC count after mobilization: P<0.001, 95% CI 0.07(0.05 - 0.09), collection cycle amount [ P<0.001, 95% CI 0.00(0.00 - 0.00)] and postharvest total amount [ P<0.001, 95% CI 0.07(0.05 - 0.10)] were the influencing factors of the collected MNC number. Meanwhile, these factorswere also the influencing factors of the collected CD 34+ number (MNC count after mobilization: P<0.001, 95% CI 0.09(0.04 - 0.14); collection cycle amount: P = 0.003, 95% CI 0.00(0.00 - 0.00); postharvest total amount: P = 0.005, 95% CI 0.08(0.03 - 0.14)). Conclusions:The collection efficiency of peripheral blood stem cells varies greatly among individuals. The more MNC counts after mobilization, the more peripheral blood stem cells could be collected. In order to obtain high collection efficiency, it is necessary to adjust the parameters of peripheral blood MNC before collection, and pay attention to the collection circulation quantity, postharvest total amount and white membrane volume.