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.

Protective effects of cinnamaldehyde on benzene-induced immune and oxidative stress injury in mice / 军事医学

Objective To investigate the effects of cinnamaldehyde,the main active component of cinnamon,on benzene-induced immune injury in mice and the related mechanism.Methods Forty male BALB/c mice were randomly divided into the control group,model group(benzene 500 mg/kg),cinnamaldehyde low,medium and high dose groups(5,25,50 mg/kg),with 8 mice in each group.Except the control group,mice in each group were treated with benzene by intragastric administration daily to induce immune and oxidative stress damage,but the intervention group was treated with cinnamaldehyde 5 times/week for 3 weeks.After medication,peripheral blood was collected 24 h after the last gavage for blood cell count,and the changes in body weight of mice in each group were observed.The pathological structure of the spleen and thymus was observed via hematoxylin-eosin(HE)staining.Peripheral blood mononuclear cells(PBMCs)of mice were extracted and the amounts of reactive oxygen species(ROS)and ATP in mitochondria were measured.Plasma levels of malondialdehyde(MDA)were measured using the barbituric acid method,the activity of glutathione peroxidase(GSH-PX)in plasmawith the dithiodinitrobenzoic acid methodand the activity of total superoxide dismutase(SOD)in plasma using the hydroxylamine method.Results After exposure to benzene,the body weight of the model group became lower(P<0.05).The spleen and thymus were damaged,and the indexes of the spleen and thymus were decreased(P<0.05).Counts of peripheral white blood cells and lymphocyteswere decreased(P<0.05).The activities of GSH and SOD in plasma were decreased(P<0.05),but the content of MDA was increased(P<0.05).The amount of mitochondrial ROS in PBMC was increased,while the ATP content was decreased(P<0.05).The weight of mice increased after treatment with cinnamaldehyde.The spleen and thymus tissues recovered well,and the indexes of the spleen and thymus were increased(P<0.05).Counts of peripheral white blood cells and lymphocytesin the high dose cinnamaldehyde group were increased(P<0.05).The activities of GSH and SOD in plasma were increased,while the content of MDA was decreased(P<0.05).The amount of mitochondrial ROS in PBMC was decreased,but the ATP content was increased(P<0.05).Treatment with cinnamaldehyde could alleviate the damage to the mitochondrial function of PBMC induced by benzene in mice,and 50 mg/kg was the best dose(P<0.05).The therapeutic effect of cinnamaldehyde had a dose-response relationship.Conclusion Cinnamaldehyde can inhibit benzene-induced immune injury and oxidative stress injury in mice by delivering an antioxidant effect and improving mitochondrial enhancement of PBMC.

Generation of an induced pluripotent stem cell line from a patient with surfactant metabolism dysfunction carrying ABCA3 mutations / 中华实用儿科临床杂志

Objective:Induced pluripotent stem cells (iPSCs) cell lines were established using peripheral blood mononuclear cells (PBMCs) from a patient suffering from neonatal respiratory distress syndrome (NRDS) who carried Adenosine triphosphate-binding cassette transporter A3 ( ABCA3) compound heterozygous mutations. Methods:Cell experimental research.Peripheral venous blood was collected and PBMCs were isolated and cultured in vitro. PBMCs were transfected with non-integrated Sendai vector carrying reprogramming factors.The chromosome karyotypes of the established iPSCs were analyzed.Immunofluorescence and flow cytometry were used to detect pluripotency markers of stem cells and verify their differentiation potential.Sanger sequencing was performed to analyze gene mutations.In addition, short tandem repeat (STR) analysis was performed, polymerase chain reaction(PCR) and agarose gel electrophoresis were used to detect virus residual. Results:Karyotype analysis of established iPSCs cell lines showed normal diploid 46, XY karyotype.Immunofluorescence showed positive staining of stem cell pluripotency markers OCT4, SSEA4, Nanog and Sox2.Flow cytometry was used to detected stem cell pluripotency markers and showed expression of TRA-1-60, SSEA-4 and OCT4.After differentiation into all three germ layers, immunofluorescence was performed to detect ectoderm (Pax-6), mesoderm (Brachyury) and endoderm alpha-fetoprotein markers, and the results showed positive staining, which confirmed that the iPSCs had the potential to differentiate.Sanger sequencing showed c. 3997_3998del and c. 3137C>T compound heterozygous mutations.STR analysis showed they originate from PBMCs, and no Sendai virus residual was detected by PCR and agarose gel electrophoresis.Conclusions:In this study, PBMCs from patient carrying ABCA3 compound heterozygous mutations was used to establish iPSCs cell lines.The research lays a foundation for the study of pathogenesis, therapeutic drug screening and cell therapy of NRDS caused by ABCA3 gene mutations.

Targeted induction of human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum into neural stem cells / 中国组织工程研究

BACKGROUND:There are many kinds of cell media with different components,which have a great influence on cell growth.Several studies in and outside China have used serum-free media containing fetal bovine serum for in vitro amplification culture,but the use of media containing human peripheral blood serum to directionally induce human umbilical cord mesenchymal stem cells to neural stem cells and human peripheral blood serum to promote neural stem cells to differentiate into other nerve cells,so far there are few relevant studies. OBJECTIVE:To observe the feasibility of inducing human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum into neural stem cells. METHODS:(1)Human umbilical cord mesenchymal stem cells were cultured in DMEM/F-12 culture medium containing 10%human peripheral blood serum by volume fraction.Flow cytometry analysis was performed at the third passage to identify surface markers and alizarin red staining was used to detect osteogenic differentiation function.(2)The third-generation human umbilical cord mesenchymal stem cells were induced into neural stem cells using DMEM/F-12 medium containing 0.5%N2,1.5%B27,20 ng/mL basic fibroblast growth factor,and 20 ng/mL epidermal growth factor,and their surface markers were identified.(3)Well-growing human umbilical cord mesenchymal stem cell-derived neural stem cells were taken to prepare a single cell suspension.They were evenly inoculated into 96-well plates and incubated with DMEM/F-12 culture medium containing 10%human peripheral blood serum for 8 days.Then,hematoxylin-eosin staining,microtubule-associated protein 2,and glial fibrillary acidic protein immunofluorescence staining were performed to detect the differentiation of neural stem cells into other neural cells. RESULTS AND CONCLUSION:(1)Human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum grew in a spiral-like pattern and were distributed in multiple layers,with directional arrangement.The surface of human umbilical cord mesenchymal stem cells highly expressed CD44,CD105,CD29,and CD73.Cells stained with alizarin red showed a color reaction.(2)Human umbilical cord mesenchymal stem cells cultured with human peripheral blood serum could be induced to differentiate into neural stem cells,and the surface of neural stem cells was highly expressed with CD44,CD105,CD29,CD73,Nestin,NF-L,and GALC.(3)On day 8 of induced differentiation of neural stem cells,after staining with hematoxylin and eosin,it was found that the protruding protrusions were longer,with more branches and adjacent cells connected,presenting typical neural cell morphology.Immunofluorescence staining for microtubule-associated protein 2 and glial fibrillary acidic protein was positive.It is concluded that human umbilical cord mesenchymal stem cells cultured by human peripheral blood serum can be directly induced to differentiate into neural stem cells.Under the action of human peripheral blood serum,neural stem cells can differentiate into other neural cells as the culture time prolongs.

Role of non-coding RNA and exosomes in pathogenesis of gestational diabetes mellitus and their early diagnostic value / 中国组织工程研究

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.

EGFR Methylation in Peripheral Blood of Driver Gene-Positive Lung Cancer and its Clinical Correlation / 医学研究杂志

Objective To explore the methylation status of EGFR promoter region in peripheral blood of EGFR driver gene-positive lung adenocarcinoma patients and its clinical significance.Methods The methylation of EGFR promoter region in peripheral blood DNA of 32 EGFR driver gene-positive lung adenocarcinoma patients and 24healthy controls were detected by(matrix-assisted laser desorp-tion/ionisation,time-of-flight mass spectrometry,MALDI-TOF MS)technique to screen the differential methylation of CpG sites with lung cancer and analyze its correlation with clinicopathological characteristics and EGFR mutant subtype.Results The methylation levels of EGFR were significantly lower in the lung cancer group than in the normal control,EGFR#29 CpG_2,EGFR#30 CpG_8.9.10.11,CpG_25,CpG_40.41.42methylation of lung cancer group(25.7％±2.1％,4.6％±0.4％,2.8％±0.7％,4.40±0.4％)were low-er than the normal control(36.5％±4.6％,6.0％±0.6％,4.8％±0.9％,7.1％±0.4％),the difference was statistically signifi-cant(P＜0.05).No significant differences in the methylation levels within EGFR CpGs were associated with age,cancer differentiation,disease location,TNM stage with lung cancer(P＞0.05).The methylation level of EGFR#30 CpG_25 in the female group was signifi-cantly lower than that in the male group(P＜0.05),and the methylation level of EGFR#30 CpG_25 in the smoking group was higher than that in the non-smoking group(P＜0.05),the methylation level of EGFR#30 CpG_8.9.10.11 in the lymph node metastasis group was significantly lower than that in the non-lymph node metastasis(P＜0.01).There was no significant difference in methylation of EGFR between EGFR 19del and 21 L858R mutant subtypes.Conclusion Our findings suggest that EGFR methylation may be associated with the occurrence,development,gender,and smoking history of patients with EGFR driving gene-positive lung cancer,which may serve as a biomarker for the diagnosis and prognosis prediction of lung cancer.

The expression of System Xc-/ GSH / GPX4 ferroptosis pathway in peripheral blood mononuclear cells of rheumatoid arthritis patients and its effect on the secretion of inflammatory factors / 安徽医科大学学报

Objective @#To investigate the expression of genes and proteins in the peripheral blood mononuclear cell ( PBMC) cystine / glutamate antiporter system (System Xc-) / glutathione ( GSH) / glutathione peroxidase 4 ( GPX4) ferroptosis pathway and its influence on inflammatory factors in patients with rheumatoid arthritis ( RA) ．@*Methods @#30 patients with RA and 30 healthy participants were enrolled．PBMCs were isolated using Ficoll-hypaque density gradient centrifugation．The cells were categorized into the healthy control，RA，ferroptosis inhibitor，ferroptosis in- ducer group．The cell viability was checked using the cell counting kit-8 ( CCK-8) method．Intracellular Fe2 + rela- tive fluorescence intensity and reactive oxygen species (ROS) levels were detected using the FerroOrange and Di- hydroethidium (DHE) fluorescent probes，respectively．Western blot and real-time quantitative PCR (qPCR) de- tected the expression of nuclear factor erythroid 2-related factor 2 ( Nrf2 ) ，solute carrier family 7 member 11 (SLC7A11) ，GPX4 proteins and mRNA．And the flow cytometry quantified the levels of tumor necrosis factor α (TNF-α) ，Interleukin (IL) -1，and IL-6 in the supernatant of each cell group. @*Results @# Compared to the healthy control group，the RA group showed a significantly increased Fe2 + concentration and elevated ROS levels，reduced expression of Nrf2，SLC7A11 and GPX4 proteins and mRNA，and increased contents of TNF-α , IL-1 and IL-6 in PBMC supernatant，and the differences were statistically significant．The concentration of Fe2 + and ROS levels in the inhibitor group were lower than those in the RA group，the proteins expressions of Nrf2，SLC7A11 and GPX4 increased，the mRNA expressions of SLC7A11 and GPX4 increased，the content of IL-6 in the PBMC supernatant decreased but the content of TNF-α increased，and the differences were statistically significant．In contrast，the in- ducer group，when compared to the RA group，displayed increased ROS levels，reduced expression of SLC7A11 protein and mRNA and decreased expression of Nrf2 protein，and the contents of TNF-α and IL-1 in the PBMC su- pernatant increased，but the expression of GPX4 protein increased ，and the differences were statistically signifi- cant．The inducer group，compared to the RA group，showed increased cell viability，and the difference was statis- tically significant (P＜0. 000 1) ．@*Conclusion @# The presence of ferroptosis in PBMC in RA patients，inhibiting or inducing PBMC ferroptosis in RA patients，will inhibit or promote the secretion of inflammatory factors．Inhibition of PBMC ferroptosis in RA patients may be helpful in the treatment of RA.

Correlation Between Peripheral Blood Inflammatory Markers and 18F-FDG PET/CT Metabolic Parameters in Tumors / 肿瘤防治研究

Inflammatory markers in peripheral blood, such as neutrophil-lymphocyte ratio and platelet-lymphocyte ratio, can reflect the reactive hyperplasia of inflammatory cells in tumors. The metabolic parameters of 18F-FDG PET/CT are also correlated with the reactive hyperplasia of inflammatory cells in tumors. However, only a few reports exist on the relationship between tumor metabolic parameters and peripheral blood inflammatory markers. Therefore, this review starts from three aspects: tumor peripheral blood inflammatory markers, inflammatory cell reactive hyperplasia in tumors, and 18F-FDG PET/CT metabolic parameters. The correlation between 18F-FDG PET/CT metabolic parameters and peripheral blood inflammatory markers is reviewed.

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.

Role and mechanism of BRCA2 gene in immune killing of lung cancer cells / 中国免疫学杂志

Objective:To study the effect and molecular mechanism of BRCA2 gene on the killing of lung cancer by immune cells.Methods:siRNA was designed to reduce BRCA2 gene in lung cancer cells;BRCA2 expression was detected by qPCR and Western blot;cell growth was detected by MTT and CCK-8 methods;peripheral blood mononuclear cells were co-cultured with lung cancer cells,and GFP fluorescence was detected by enzyme labeling method.Killing efficiency of lung cancer cells was evaluated.Results:BRCA2 gene was expressed in moderate abundance in lung cancer cells A549 and H1299,and there was no significant differ-ence compared with lung epithelial cells BEAS-2B(P>0.05).When the BRCA2 gene in A549 and H1299 cells was successfully knocked down,the cell proliferation rate was significantly increased compared with control group;the killing efficiency of peripheral blood mononuclear cells to lung cancer cells A549 and H1299 were significantly higher than that of the control group;the expression of ATM,RAD51 and RAD50 were significantly reduced,while the expression of P53 protein was 7.2 times of the control group.Con-clusion:After knockdown of BRCA2 gene,peripheral blood monocytes are more effective in killing lung cancer cells.Intervention of BRCA2 and monocytes can synergistically inhibit lung cancer and regulate the expression of ATM signaling pathway molecules.

Analysis of peripheral immune cell typing in breast cancer lung metastasis model of miR-155 knockout mice / 中国免疫学杂志

Objective:To establish a mouse model of breast cancer lung metastasis with miR-155 knockout(miR155-/-)mice,and to compare the difference of peripheral blood immune cell typing between miR155-/-mice and C57BL/6J wide-type(WT)mice.Methods:Bioinformatics analysis was used to explore the expression level of miR-155 in breast cancer tissues and peripheral serum,and its relationship with prognosis.Mouse model of lung metastasis of breast cancer was established by tail vein injection;peripheral blood was collected for flow cytometry,and the immune cell typing was analyzed;the lung tissues were collected for immunohisto-chemical detection to observe the tumor metastasis.Results:Percentage of T lymphocytes and monocytes in peripheral blood of miR155-/-mice was significantly decreased compared with WT mice(P<0.05),percentage of myeloid inhibitory cells(MDSCs)was increased significantly(P<0.05),in which the proportion of monocyte subsets(M-MDSC)was significantly decreased(P<0.05),while the proportion of granulocyte subsets(G-MDSC)was significantly increased(P<0.05).In lung metastasis model of breast can-cer,percentage of T lymphocytes in peripheral blood of miR155-/-mice was significantly higher compared with WT mice,while per-centage of NK cells was decreased significantly(P<0.05),percentage of neutrophil was significantly decreased(P<0.001),propor-tion of Th cells in T lymphocytes was significantly decreased(P<0.05),proportion of M-MDSCs was significantly decreased(P<0.01),while proportion of G-MDSCs was significantly increased(P<0.01).Conclusion:Deletion of miR-155 gene leads to significant differences in peripheral immune cell typing,making mice more susceptible to lung metastasis of breast cancer.

Screening and analysis of ferroptosis related genes in peripheral blood mononuclear cells of patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective:To analyze the differentially expressed genes in PBMCs of patients with systemic lupus erythematosus (SLE) by bioinformatics methods screening and analyzing the key genes related to ferroptosis, and explore the possible mechanism of ferroptosis involved in the pathogenesis of SLE at the transcription level.Methods:The data sets and samples of healthy people (HC) and SLE patients who met the screening criteria were retrieved from the Gene Expression Omnibus (GEO), a sub-database of the National Center for Biotechnology Information (NCBI). The differentially expressed genes, GO enrichment analysis and KEGG pathway enrichment analysis were analyzed by GEO2R, R language and related software packages. The protein interaction network (PPI) of differential genes was analyzed by STRING, Cytoscape and other tools to explore the key genes and pathways. In addition, real-time quantitative reverse transcription PCR (RT-qPCR) was used to verify the expression of key genes. Mann-Whitney U test was used to compare the expression of key genes in PBMCs between the two groups. Spearman rank correlation analysis was used to explore the relationship between SLE disease activity and the level of key genes. Results:Six data sets were included in this study. A total of 166 genes related to ferroptosis were differentially expressed between SLE and HC groups. The differential genes were specifically expressed in alveolar macrophages, neutrophils, CD49 + cells and CD31 + cells. GO enrichment analysis and KEGG pathway enrichment analysis showed that the differentially expressed genes were mainly involved in multiple signaling pathways closely related to SLE, such as oxidative stress response, infection and TNF signaling pathway. Hub genes screened by different algorithms all suggested RELA as a key gene, and RT-qPCR confirmed that compared with the RELA gene expression level in the HC group [0.75(0.37,1.13)], the expression level in SLE group [2.02 (1.19,4.06)] was increased, the difference was statistically significant ( Z=-3.08, P=0.002), and was positively correlated with the corresponding SLEDAI score of SLE samples ( r=0.52, P=0.019). Conclusion:The ferroptosis of many immune cells, including alveolar macrophages and CD49 + NK cells, is involved in the pathogenesis of SLE. RELA may be involved in the ferroptosis of PBMCs in SLE through the NF-κB pathway.

Establishing a dose-response curve for 60Co γ-ray irradiation-induced micronuclei in human peripheral blood cells using artificial intelligence-based color recognition / 中华放射医学与防护杂志

Objective:To identify micronuclei through the cytochalasin B blocking micronucleus method-based assay using scanning microscope, combined with the slide scanning software Metafer 4 and, accordingly, to establish a dose-response relationship between the dose of 60Co γ-rays and the frequency of micronuclei in human peripheral blood lymphocytes using artificial intelligence-based color recognition. Methods:Blood samples were collected from four healthy individuals (two men and two women) and were then exposed to varying doses of 60Co γ-ray radiation (0, 0.25, 0.5, 1, 2, 3, 4, 5 Gy) at a dosage rate of 0.74 Gy/min. Micronucleus slides were prepared as per the GBZ 128-2023 standard. The numbers of binuclear cells and micronuclei were recorded using an artificial intelligence-based color recognition analysis system. The dose-response curve was determined through fitting using the CABAS software. Then, the doses to both independent samples were estimated based on the curve. Results:Within a dose range of 0 to 5 Gy, the fitted micronucleus dose-response curve aligned with a quadratic polynomial model, with a regression equation of y = 0.032 1 D2+ 0.023 7 D+ 0.012 7 ( D denoting the dose, correlation coefficient R2=0.998). The dose estimations from the validation samples closely corresponded to the actual irradiation doses. Conclusions:Establishing the micronucleus dose-response curve provides a feasible method and basis for the rapid and accurate estimation of radiation biological doses in laboratory automation.

Platelet Rich Fibrin Matrix (PRFM) and Peripheral Blood Mesenchymal Stem Cells (PBMSCs) in the management of intraosseous defects - A randomized clinical trial

Abstract A combination of peripheral blood mesenchymal stem cells (PBMSCs) and platelet rich fibrin matrix (PRFM) could be a probable periodontal regenerative material with the synergy of the added benefits of each material. Objective This randomized controlled clinical trial aimed to evaluate the regenerative capacity of supercell (PRFM and PBMSCs) compared with that of PRFM alone in human periodontal mandibular intraosseous defects (IOD). Methodology This study included 17 patients of both sexes (12 men, 5 women) aged 30-55 years (mean age = 37.7±4.4 years) who fulfilled the inclusion criteria (radiographic and clinical evaluation for bilateral IOD with probing pocket depth (PPD ≥ 6 mm). A split-mouth design was used in each patient. A total of 34 sites in the mandibular arch randomly received PRFM alone + open flap debridement (OFD) [Control sites] or supercell (PRFM+PBMSCs) + OFD [Test sites]. The clinical parameters plaque index (PI), gingival index (GI), PPD, clinical attachment level (CAL), and in the radiographic parameters; defect depth (DD) and defect fill percentage (DFP) were recorded at baseline, 3 and 6 months postoperatively. Early wound healing index (EHI) was used at 1 week to assess wound healing ability. Results At 6 months, radiographic parameters revealed significant reduction in DD (P<0.001) and significant DFP values in the test group compared with the control group. The supercell showed significant improvement in PPD and CAL at the end of 6 months (P<0.001). EHI scores at 1 week showed no statistically significant difference between the test and control groups. Conclusion Supercell can be considered a regenerative material in the treatment of periodontal IODs.

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.

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 .

Improvement of inflammatory cytokines level in peripheral blood, anxiety and depressive symptoms in patients with pulpitis by root canal therapy / 口腔疾病防治

Objective@# To investigate the effect of root canal therapy (RCT) on inflammatory cytokines level in peripheral blood, anxiety, and depression in patients with pulpitis.@*Methods @#A total of 155 patients with pulpitis admitted to the Stomatology Hospital of the Fourth Military Medical University from June 2021 to June 2022 were treated with root canal therapy. Another 155 persons who received health examinations during the same period were selected as the control group. The Generalized Anxiety Disorder-7 (GAD-7) scores and the Patient Health Questionnaire-9 (PHQ-9) scores of the two groups were compared. The GAD-7, PHQ-9 and pain scores of the test group before treatment and 3 and 6 weeks after treatment were compared. Pain was assessed with a visual analog scale (VAS). Inflammatory cytokine [interleukin-8 (IL-8), interleukin-1 β (IL-1β), tumor necrosis factor-α (TNF-α), c-reactive protein (CRP)] levels in the test group before treatment and 3 and 6 weeks after treatment were compared. @*Results @#The GAD-7 and PHQ-9 scores in the test group were higher than those in the control group before treatment (P<0.05). The GAD-7 and PHQ-9 scores in the test group at 3 and 6 weeks after treatment were lower than those before treatment (P<0.05); there was no significant difference between the GAD-7 and PHQ-9 scores at 3 and 6 weeks after treatment(P>0.05). The pain scores of the experimental group at 3 and 6 weeks after treatment were lower than those before treatment (P<0.05), and the pain scores 6 weeks after treatment were lower than those at 3 weeks after treatment (P<0.05). The levels of IL-8, IL-1β, TNF-α and CRP in the peripheral blood of the experimental group were lower 3 and 6 weeks after treatment than before (P<0.05), and the levels of IL-8 and IL-1β in the peripheral blood at 6 weeks after treatment were significantly lower than at 3 weeks after treatment (P<0.05). The levels of TNF-α and CRP in the peripheral blood at 6 weeks after treatment were not significantly different from those at 3 weeks after treatment (P>0.05).@*Conclusion @#The peripheral blood of patients with pulpitis has a high level of inflammatory cytokines, and the patients suffer from obvious anxiety and depression. Root canal therapy can relieve their anxiety and depression by reducing their level of inflammatory cytokines in peripheral blood.

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.

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.