Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnosti...Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnostic value for hepatitis B virus-associated hepatocellular carcinoma(HBV-HCC). Methods: We enrolled 197 patients, 111 with HBV-HCC, 51 with chronic hepatitis B(CHB), and 35 healthy controls(HCs). The methylation status of GLDC promoter in peripheral mononuclear cells(PBMCs) was identified by methylation specific polymerase chain reaction(MSP). The mRNA expression was examined using real-time quantitative polymerase chain reaction(q PCR). Results: The methylation frequency of the GLDC promoter was significantly lower in HBV-HCC patients(27.0%) compared to that in CHB patients(68.6%) and HCs(74.3%)( P < 0.001). The methylated group had lower alanine aminotransferase level( P = 0.035) and lower rates of tumor node metastasis(TNM) Ⅲ/Ⅳ( P = 0.043) and T3/T4( P = 0.026). TNM stage was identified to be an independent factor for GLDC promoter methylation. GLDC mRNA levels in CHB patients and HCs were significantly lower than those in HBV-HCC patients( P = 0.022 and P < 0.001, respectively). GLDC mRNA levels were significantly higher in HBV-HCC patients with unmethylated GLDC promoters than those with methylated GLDC promoters( P = 0.003). The diagnostic accuracy of alpha-fetoprotein(AFP) combined with GLDC promoter methylation for HBV-HCC was improved compared with that of AFP alone(AUC: 0.782 vs. 0.630, P < 0.001). In addition, GLDC promoter methylation was an independent predictor for overall survival of HBV-HCC patients( P = 0.038). Conclusions: The methylation frequency of GLDC promoter was lower in PBMCs from HBV-HCC patients than that from patients with CHB and HCs. The combination of AFP and GLDC promoter hypomethylation significantly improved the diagnostic accuracy of HBV-HCC.展开更多
Objective:To compare the effect of human chorionic gonadotropin(hCG)-producing peripheral blood mononuclear cells(PBMCs)and PBMCs activated by hCG in vitro and expressions of related immune genes in mouse implantation...Objective:To compare the effect of human chorionic gonadotropin(hCG)-producing peripheral blood mononuclear cells(PBMCs)and PBMCs activated by hCG in vitro and expressions of related immune genes in mouse implantation.Methods:hCG-producing PBMCs(transfected PBMC)and PBMCs activated by hCG in vitro were introduced into isolated mouse endometrial cells,while cell cultures were divided into four groups:the control,PBMC,transfected,and activated PBMC groups.The expression of studied genes(IL-1β,IL-6,Lif,and Vegf)was evaluated and blastocyst attachment on the cocultured cells(isolated endometrial cells and PBMC cells)was monitored in all four groups.Results:Data showed that expression decreased in the PBMC group compared to the treated PBMC(transfected and activated PBMCs)and increased in transfected PBMC compared to the activated PBMC.Attachment and migration of blastocysts were dramatically enhanced in the transfected PBMC group compared to the activated PBMC group(P<0.05).Conclusions:Use of hCG-producing PBMCs(transfected PBMC)has more influence on endometrial receptivity.展开更多
AIM To investigate disease-specific gene expression profiles of peripheral blood mononuclear cells(PBMCs) from Crohn's disease(CD) patients in clinical remission.METHODS Patients with CD in clinical remission or w...AIM To investigate disease-specific gene expression profiles of peripheral blood mononuclear cells(PBMCs) from Crohn's disease(CD) patients in clinical remission.METHODS Patients with CD in clinical remission or with very low disease activity according to the Crohn's disease activity index were genotyped regarding nucleotidebinding oligomerization domain 2(NOD2),and PBMCs from wild-type(WT)-NOD2 patients,patients with homozygous or heterozygous NOD2 mutations and healthy donors were isolated for further analysis.The cells were cultured with vitamin D,peptidoglycan(PGN) and lipopolysaccharide(LPS) for defined periods of time before RNA was isolated and subjected to microarray analysis using Clariom S assays and quantitative realtime PCR.NOD2-and disease-specific gene expression profiles were evaluated with repeated measure ANOVA by a general linear model.RESULTS Employing microarray assays,a total of 267 genes were identified that were significantly up-or downregulated in PBMCs of WT-NOD2 patients,compared to healthy donors after challenge with vitamin D and/or a combination of LPS and PGN(P < 0.05;threshold:≥ 2-fold change).For further analysis by real-time PCR,genes with known impact on inflammation and immunity were selected that fulfilled predefined expression criteria.In a larger cohort of patients and controls,a disease-associated expression pattern,with higher transcript levels in vitamin D-treated PBMCs from patients,was observed for three of these genes,CLEC5 A(P < 0.030),lysozyme(LYZ;P < 0.047) and TREM1(P < 0.023).Six genes were found to be expressed in a NOD2-dependent manner(CD101,P < 0.002;CLEC5 A,P < 0.020;CXCL5,P < 0.009;IL-24,P < 0.044;ITGB2,P < 0.041;LYZ,P < 0.042).Interestingly,the highest transcript levels were observed in patients with heterozygous NOD2 mutations.CONCLUSION Our data identify CLEC5 A and LYZ as CD-and NOD2-associated genes of PBMCs and encourage further studies on their pathomechanistic roles.展开更多
Antipsychotics may prolong or retain telomere length,affect mitochondrial function,and then affect the metabolism of nerve cells.To validate the hypothesis that antipsychotics can prolong telomere length after oxidati...Antipsychotics may prolong or retain telomere length,affect mitochondrial function,and then affect the metabolism of nerve cells.To validate the hypothesis that antipsychotics can prolong telomere length after oxidative stress injury,leukocytes from healthy volunteers were extracted using Ficoll-Histopaque density gradient.The mononuclear cells layer was resuspended in cell culture medium.Oxidative stress was induced with hydrogen peroxide in cultured leukocytes.Four days later,leukocytes were treated with aripiprazole,haloperidol or clozapine for 7 days.Real-time PCR revealed that treatments with aripiprazole and haloperidol increased the telomere length by 23%and 20%in peripheral blood mononuclear cells after acute oxidative stress injury.These results suggest that haloperidol and aripiprazole can reduce the damage to telomeres induced by oxidative stress.The experiment procedure was approved by the Ethics Committee of Faculty of Medicine of the University of Sao Paulo(FMUSP/CAAE approval No.52622616.8.0000.0065).展开更多
To explore changes of toll-like receptor (TLR) 2,4 in peripheral blood mononuclear cells (PBMC) in acute abdomen patients with systemic inflammatory response syndrome (SIRS) and their significance.Methods A clinical s...To explore changes of toll-like receptor (TLR) 2,4 in peripheral blood mononuclear cells (PBMC) in acute abdomen patients with systemic inflammatory response syndrome (SIRS) and their significance.Methods A clinical study was done on 103 patients of which 65 were with SIRS.The mRNA expression of TLR2,4 were detected by RT-PCR;the expression of TNF-α and IL-6 were observed by ELISA;the correlation between TLR2,4 mRNA,the level of TNF-α and IL-6,and the clinical course was evaluated.Results TLR2 mRNA ,TNF-α and IL-6 were upregulated markedly on the first day of hospitalization,then decreased gradually;TLR2 mRNA maintained on high level till the 5th day.The expression of TLR2,4 mRNA was positive correlated with the level of TNF-α and IL-6,and the length of stay.TLR2,4 mRNA expression increased in patients with multiple organ failure.Conclusion In actue abdomen patients with SIRS,the expression of TLR2,4 of PBMC increased markedly,indicating its improtant role in the pathogenesis of SIRS.4 refs,2 figs,2 tabs.展开更多
Objective:This study aimed to investigate the immune response of a pregnant woman who recovered from the coronavirus disease 2019(COVID_(R)S)by using single-cell transcriptomic profiling of peripheral blood mononuclea...Objective:This study aimed to investigate the immune response of a pregnant woman who recovered from the coronavirus disease 2019(COVID_(R)S)by using single-cell transcriptomic profiling of peripheral blood mononuclear cells(PBMCs)and to analyze the properties of different immune cell subsets.Methods:PBMCs were collected from the COVID_(R)S patient at 28 weeks of gestation,before a cesarean section.The PBMCs were then analyzed using single-cell RNA sequencing.The transcriptional profiles of myeloid,T,and natural killer(NK)cell subsets were systematically analyzed and compared with those of healthy pregnant controls from a published single-cell RNA sequencing data set.Results:We identified major cell types such as T cells,B cells,NK cells,and myeloid cells in the PBMCs of our COVID_(R)S patient.The increase of myeloid and B cells and decrease of T cells and NK cells in the PBMCs in this patient were quite distinct compared with that in the control subjects.After reclustering and Augur analysis,we found that CD16 monocytes and mucosal-associated invariant T(MAIT)cells were mostly affected within different myeloid,T,and NK cell subtypes in our COVID_(R)S patient.The proportion of CD16 monocytes in the total myeloid population was increased,and the frequency of MAIT cells in the total T and NK cells was significantly decreased in the COVID-RS patient.We also observed significant enrichment of gene sets related to antigen processing and presentation,T-cell activation,T-cell differentiation,and tumor necrosis factor superfamily cytokine production in CD16 monocytes,and enrichment of gene sets related to antigen processing and presentation,response to type II interferon,and response to virus in MAIT cells.Conclusion:Our study provides a single-cell resolution atlas of the immune gene expression patterns in PBMCs from a COVID_(R)S patient.Our findings suggest that CD16-positive monocytes and MAIT cells likely play crucial roles in the maternal immune response against severe acute respiratory syndrome coronavirus 2 infection.These results contribute to a better understanding of the maternal immune response to severe acute respiratory syndrome coronavirus 2 infection and may have implications for the development of effective treatments and preventive strategies for the coronavirus disease 2019 in pregnant women.展开更多
Vogt-Koyanagi-Harada disease(VKH)is a rare autoimmune disease characterized by diffuse and bilateral uveitis,alopecia,tinnitus,hearing loss,vitiligo and headache.The transcriptional expression pattern of peripheral bl...Vogt-Koyanagi-Harada disease(VKH)is a rare autoimmune disease characterized by diffuse and bilateral uveitis,alopecia,tinnitus,hearing loss,vitiligo and headache.The transcriptional expression pattern of peripheral blood mononuclear cells(PBMC)in VKH remains largely unknown.In this study,mRNA sequencing was conducted in PBMC from VKH patients with active uveitis before treatment(n=7),the same patients after prednisone combined with cyclosporine treatment(n=7)and healthy control subjects strictly matched with gender and age(n=7).We found 118 differentially expressed genes(DEGs)between VKH patients and healthy control subjects,and 21 DEGs between VKH patients before and after treatment.TRIB1 was selected as a potential biomarker to monitor the development of VKH according to the mRNA sequencing.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were performed to predict the possible biological functions and signaling pathways of DEGs.Neutrophil degranulation,peptidase regulator activity,secretory granule membrane,cellular response to peptide,growth factor binding and cell projection membrane were enriched as GO annotations of DEGs.Arachidonic acid metabolism and mitogen-activated protein kinase(MAPK)signaling pathway were potential signaling pathways involved in pathogenesis and drug response of VKH.A protein–protein interaction(PPI)network was constructed by STRING,and colony stimulating factor 1 receptor(CSF1R)was identified as the hubgene of all DEGs by Cytoscape.The cell type presumed to contribute to the aberrant expression of DEGs was analyzed with the use of publicly available single-cell sequencing data of PBMC from a healthy donor and single-cell sequencing dataset of monocytes from VKH patients.Our findings may help to decipher the underlying cellular and molecular pathogenesis of VKH and may lead novel therapeutic applications.展开更多
Intrahepatic cholangiocarcinoma(ICC)is the second most common liver cancer.Chemotherapy remains the main therapeutic strategy for advanced ICC patients,but chemosensitivity varies individually.Here,we applied cytometr...Intrahepatic cholangiocarcinoma(ICC)is the second most common liver cancer.Chemotherapy remains the main therapeutic strategy for advanced ICC patients,but chemosensitivity varies individually.Here,we applied cytometry by time-of-flight(CyTOF)to establish the immune profile of peripheral blood mononuclear cells(PBMCs)on the single-cell level at indicated time points before,during,and after chemotherapy.Multiplex immunofluorescence staining was applied to examine the spatial distribution of certain immune clusters.Tissue microarrays(TMAs)were used for prognostic evaluation.A total of 20 ICC patients treated with gemcitabine(GEM)were enrolled in our study,including eight cases with good response(R)and 12 cases with non-response(NR).Tremendous changes in PBMC composition,including an increased level of CD4/CD8 double-positive T cells(DPT),were observed after chemotherapy.Patients with higher level of CD4^(+)CD45RO^(+)CXCR3^(+)T cells before treatment had a favorable response to chemotherapy.Our study identified a positive correlation between the percentage of T cell subpopulations and clinical response after chemotherapy,which suggests that it is practical to predict the potential response before treatment by evaluating the proportions of the cell population in PBMCs.展开更多
In recent years,several studies have reported positive outcomes of cell-based therapies despite insufficient engraftment of transplanted cells.These findings have created a huge interest in the regenerative potential ...In recent years,several studies have reported positive outcomes of cell-based therapies despite insufficient engraftment of transplanted cells.These findings have created a huge interest in the regenerative potential of paracrine factors released from transplanted stem or progenitor cells.Interestingly,this notion has also led scientists to question the role of proteins in the secretome produced by cells,tissues or organisms under certain conditions or at a particular time of regenerative therapy.Further studies have revealed that the secretomes derived from different cell types contain paracrine factors that could help to prevent apoptosis and induce proliferation of cells residing within the tissues of affected organs.This could also facilitate the migration of immune,progenitor and stem cells within the body to the site of inflammation.Of these different paracrine factors present within the secretome,researchers have given proper consideration to stromal cell-derived factor-1(SDF1)that plays a vital role in tissue-specific migration of the cells needed for regeneration.Recently researchers recognized that SDF1 could facilitate site-specific migration of cells by regulating SDF1-CXCR4 and/or HMGB1-SDF1-CXCR4 pathways which is vital for tissue regeneration.Hence in this study,we have attempted to describe the role of different types of cells within the body in facilitating regeneration while emphasizing the HMGB1-SDF1-CXCR4 pathway that orchestrates the migration of cells to the site where regeneration is needed.展开更多
Objective To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem ce...Objective To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem cells (PBSCs), and furthermore, to understand the possible causes of this response. Methods We used MTT assay to detect the survival rate of PBMCs, CBMCs and PBSCs after treated by ALA-PDT under the optimum experiment conditions with U937 as control; Annexin V-FITC/PI was used to detect the pattern of cell death induced by ALA-PDT. By using flow cytometry, we detected intracellular PpIX fluorescence intensity. Results After ALA-PDT treatment the survival rate of PBMCs had no significant change; however in PBSCs and CBMCs, the survival rate reduced to 70%, and the survival rate of leukemia cell U937 was the lowest, about 30%. After incubation with ALA, except for PBMCs, intracellular PpIX fluorescence intensity of the other two kinds of normal haemocytes and U937 increased obviously. These results combined with the flow cytometry suggested that the main pattern of cell death here was apoptosis. Conclusion Under the optimum experiment conditions, ALA-PDT has a slight effect on normal haemocytes but excellent depletions of leukemia cells. Therefore, it can effectively purify autologous bone marrow or stem cell grafts.展开更多
AIM To explore the exact interaction between Notch and transforming growth factor(TGF)-β signaling in liver fibrosis. METHODS We established a rat model of liver fibrosis induced by concanavalin A. Peripheral blood m...AIM To explore the exact interaction between Notch and transforming growth factor(TGF)-β signaling in liver fibrosis. METHODS We established a rat model of liver fibrosis induced by concanavalin A. Peripheral blood mononuclear cells(PBMCs) were isolated from the modeled rats, and cultured with γ-secretase inhibitor DAPT and TGF-β inhibitor for 24 h. The m RNA levels of Notch and TGF-β signaling were detected by quantitative real-time polymerase chain reaction. Expression of Notch and TGF-β proteins was analyzed by western blotting.RESULTS Compared to control rats, Notch and TGF-β signaling was activated in PBMCs of model rats. Administration of DAPT and TGF-β inhibitor suppressed Notch and TGF-β signal transducer in PBMCs of model rats. DAPT reduced m RNA and protein expression of TGF-β signaling, such as TGF-β1 and Smad3. TGF-β inhibitor also downregulated Notch1, Hes1 and Hes5, and m RNA and protein expression of the Notch signaling pathway.CONCLUSION Notch and TGF-β signaling play a role in liver fibrosis. TGF-β signaling upregulates Notch signaling, which promotes TGF-β signaling.展开更多
AIM:To investigate the presence of mixed infection and discrepancy between hepatitis C virus(HCV) genotypes in plasma,peripheral blood mononuclear cells(PBMCs),and liver biopsy specimens.METHODS:From September 2008 up...AIM:To investigate the presence of mixed infection and discrepancy between hepatitis C virus(HCV) genotypes in plasma,peripheral blood mononuclear cells(PBMCs),and liver biopsy specimens.METHODS:From September 2008 up to April 2009,133 patients with chronic hepatitis C referred to Firouzgar Hospital for initiation of an antiviral therapy were recruited in the study.Five milliliters of peripheral blood was collected from each patient and liver biopsy was performed in those who gave consent or had indications.HCV genotyping was done using INNO-LiPATM HCV in serum,PBMCs,and liver biopsy specimens and then conf irmed by sequencing of 5'-UTR fragments.RESULTS:The mean age of patients was 30.3 ± 17.1 years.Multiple transfusion was seen in 124(93.2%) of patients.Multiple HCV genotypes were found in 3(2.3%) of 133 plasma samples,9(6.8%) of 133 PBMC samples,and 8(18.2%) of 44 liver biopsy specimens.It is notable that the different genotypes found in PBMCs were not the same as those found in plasma and liver biopsy specimens.CONCLUSION:Our study shows that a signif icant proportion of patients with chronic hepatitis C are affected by multiple HCV genotypes which may not be detectable only in serum of patients.展开更多
Glaucoma is a neurodegenerative disease in which optic nerve damage and visual field defects occur.It is a leading cause of irreversible blindness.Its pathogenesis is largely unknown although several risk factors have...Glaucoma is a neurodegenerative disease in which optic nerve damage and visual field defects occur.It is a leading cause of irreversible blindness.Its pathogenesis is largely unknown although several risk factors have been identified,with an increase in intraocular pressure being the main one.Lowering of intraocular pressure is the only treatment available.Open-angle glaucoma is the most common form of the condition,accounting for~90%of all cases of glaucoma,with primary open-angle glaucoma and exfoliation glaucoma being the most frequent types.There are strong indications that microRNAs play important roles in the pathogenesis of primary open-angle glaucoma.Most of the recent studies reviewed had performed microRNA profiling in aqueous humor from glaucoma patients compared to controls who were chiefly cataract patients.A very large number of microRNAs were dysregulated but with limited overlap between individual studies.MiRNAs in aqueous humor that could be possible targets for therapeutic intervention are miR-143-3p,miR-125b-5p,and miR-1260b.No ove rlap of findings occurred within the dysregulated miRNAs for blood plasma,blood serum,peripheral blood mononuclear cells,and tears of primary open-angle glaucoma patients.Seve ral impo rtant limitations were identified in these studies.Further studies are warranted of mic roRNA expression in aqueous humor and blood samples of primary open-angle glaucoma patients in the early stages of the disease so that validated biomarkers can be identified and treatment initiated.In addition,whether modifying the levels of specific microRNAs in aqueous humor or tears has a beneficial effect on intraocular pressure and ophthalmic examination of the eyes should be investigated using suitable animal models of glaucoma.展开更多
Objective To evaluate the methylation status at CpG site -55 in the interferon-gamma (IFN-γ) gene promoter and its effect on IFN-γ expression in chronic hepatitis B. Method The authors recruited 30 patients with HBe...Objective To evaluate the methylation status at CpG site -55 in the interferon-gamma (IFN-γ) gene promoter and its effect on IFN-γ expression in chronic hepatitis B. Method The authors recruited 30 patients with HBeAg-positive chronic hepatitis B (CHB), 30 HBeAg-negative CHB patients, and 30 healthy blood donors. Pyrosequencing was used to determine the methylation status at CpG site -55 in the IFN-γ gene promoter following bisulfite treatment of DNA in peripheral blood mononuclear cells (PBMCs). The expression of IFN-γ was analyzed by real-time RT-PCR and ELISA. HBV DNA in PBMCs was detected by nested PCR. Results The methylation level at CpG site -55 in the IFN-γ gene promoter was significantly increased, resulting in subsequent down-regulation of the expression of this cytokine in CHB. The methylation level at CpG site -55 was significantly higher in HBeAg-positive patients than in HBeAg-negative ones (P<0.01) and was also significantly higher in PBMCs from HBV DNA-positive patients than from HBV DNA-negative ones (P<0.01); the methylation level at CpG site -55 was positively correlated with the amount of HBV DNA in serum (P<0.01). Conclusion IFN-γ gene expression appears to be regulated by methylation of the IFN-γ gene promoter in CHB; the methylation level at CpG site -55 is associated with HBV infection.展开更多
In order to increase the positive detection rate of HCV RNA in the patients with chronic hepatitis C , RT PCR was used to synchronously detect HCV RNA in the plasma and peripheral blood mononuclear cells of 583 CHC pa...In order to increase the positive detection rate of HCV RNA in the patients with chronic hepatitis C , RT PCR was used to synchronously detect HCV RNA in the plasma and peripheral blood mononuclear cells of 583 CHC patients with a continuously elevated level of ALT for more than one year. The results showed that the positive detection rate of HCV RNA in the plasma of the CHC patients was 19.2 %, while 24.5 % in PBMC. It was demonstrated that the positive detection rate for HCV RNA in PBMC was obviously higher than that detected in plasma. To synchronously detect HCV RNA in PBMC by using RT PCR can increase the positive detection rate of HCV RNA in the CHC patients.展开更多
Although widely applied in treating hematopoietic malignancies,transplantation of hematopoietic stem/progenitor cells(HSPCs)is impeded by HSPC shortage.Whether circulating HSPCs(cHSPCs)in steady-state blood could be u...Although widely applied in treating hematopoietic malignancies,transplantation of hematopoietic stem/progenitor cells(HSPCs)is impeded by HSPC shortage.Whether circulating HSPCs(cHSPCs)in steady-state blood could be used as an alternative source remains largely elusive.Here we develop a three-dimensional culture system(3DCS)including arginine,glycine,aspartate,and a series of factors.Fourteen-day culture of peripheral blood mononuclear cells(PBMNCs)in 3DCS led to 125-and 70-fold increase of the frequency and number of CD34+cells.Further,3DCS-expanded cHSPCs exhibited the similar reconstitution rate com-pared to CD34+HSPCs in bone marrow.Mechanistically,3DCS fabricated an immunomodulatory niche,secreting cytokines as TNF to support cHSPC survival and proliferation.Finally,3DCS could also promote the expansion of cHSPCs in patients who failed in HSPC mobilization.Our 3DCS successfully expands rare cHSPCs,providing an alternative source for the HSPC therapy,particularly for the patients/donors who have failed in HSPC mobilization.展开更多
BACKGROUND Occult hepatitis C infection(OCI)is characterized by the presence of hepatitis C virus(HCV)RNA in the liver,peripheral blood mononuclear cells(PBMC)and/or ultracentrifuged serum in the absence of detectable...BACKGROUND Occult hepatitis C infection(OCI)is characterized by the presence of hepatitis C virus(HCV)RNA in the liver,peripheral blood mononuclear cells(PBMC)and/or ultracentrifuged serum in the absence of detectable HCV-RNA in serum.OCI has been described in several categories of populations including hemodialysis patients,patients with a sustained virological response,immunocompromised individuals,patients with abnormal hepatic function,and apparently healthy subjects.AIM To highlight the global prevalence of OCI.METHODS We performed a systematic and comprehensive literature search in the following 4 electronic databases PubMed,EMBASE,Global Index Medicus,and Web of Science up to 6th May 2021 to retrieve relevant studies published in the field.Included studies were unrestricted population categories with known RNA status in serum,PBMC,liver tissue and/or ultracentrifuged serum.Data were extracted independently by each author and the Hoy et al tool was used to assess the quality of the included studies.We used the random-effect meta-analysis model to estimate the proportions of OCI and their 95%confidence intervals(95%CI).The Cochran's Q-test and the I2 test statistics were used to assess heterogeneity between studies.Funnel plot and Egger test were used to examine publication bias.R software version 4.1.0 was used for all analyses.RESULTS The electronic search resulted in 3950 articles.We obtained 102 prevalence data from 85 included studies.The pooled prevalence of seronegative OCI was estimated to be 9.61%(95%CI:6.84-12.73)with substantial heterogeneity[I^(2)=94.7%(95%CI:93.8%-95.4%),P<0.0001].Seropositive OCI prevalence was estimated to be 13.39%(95%CI:7.85-19.99)with substantial heterogeneity[I^(2)=93.0%(90.8%-94.7%)].Higher seronegative OCI prevalence was found in Southern Europe and Northern Africa,and in patients with abnormal liver function,hematological disorders,and kidney diseases.Higher seropositive OCI prevalence was found in Southern Europe,Northern America,and Northern Africa.CONCLUSION In conclusion,in the present study,it appears that the burden of OCI is high and variable across the different regions and population categories.Further studies on OCI are needed to assess the transmissibility,clinical significance,long-term outcome,and need for treatment.展开更多
The severe acute respiratory syndrome coronavirus 2(SARS-CovV-2)infection is associated with a hyperinflammatory state and lymphocytopenia,a hallmark that appears as both signature and prognosis of disease severity ou...The severe acute respiratory syndrome coronavirus 2(SARS-CovV-2)infection is associated with a hyperinflammatory state and lymphocytopenia,a hallmark that appears as both signature and prognosis of disease severity outcome.Although cytokine storm and a sustained inflammatory state are commonly associated with immune cell depletion,it is still unclear whether direct SARS-Cov-2 infection of immune cells could also play a role in this scenario by harboring viral replication.We found that monocytes,as well as both B and T lymphocytes,were susceptible to SARS-Cov-2 infection in vitro,accumulating double-stranded RNA consistent with viral RNA replication and ultimately leading to expressive T cell apoptosis.In addition,flow cytometry and immunofluorescence analysis revealed that SARS-Cov-2 was frequently detected in monocytes and B lymphocytes from coronavirus disease 2019(CoVID-19)patients.The rates of SARS-Cov-2-infected monocytes in peripheral blood mononuclear cells from CoVID-19 patients increased over time from symptom onset,with SARS-CoV-2-positive monocytes,B cells,and CD4+T lymphocytes also detected in postmortem lung tissue.These results indicated that SARS-CoV-2 infection of blood-circulating leukocytes in covID-19 patients might have important implications for disease pathogenesis and progression,immune dysfunction,and virus spread within the host.展开更多
Aim:Used as a palliative therapy for unresectable liver cancer,radiofrequency ablation(RFA)is associated with the induction of immunological responses.Here,we show strong evidence of tumor-specifi c peripheral blood m...Aim:Used as a palliative therapy for unresectable liver cancer,radiofrequency ablation(RFA)is associated with the induction of immunological responses.Here,we show strong evidence of tumor-specifi c peripheral blood mononuclear cells(PBMCs)12 months after RFA.Methods:Three patients with colorectal cancer(CRC)metastases to the liver and two patients with primary hepatocellular carcinoma(HCC)were enrolled in this study.PBMC,isolated 12 months after RFA,were stimulated with normal and tumor tissue lysate.Interferon gamma secretion was evaluated by fl ow cytometry and indirectly,by luciferase assay for adenylate kinase activity in PBMC-stimulated lysates of target cells.Baseline data were detected before RFA and 4 weeks after treatment.Results:Two CRC patients and one HCC patient had recurrence-free survival.One patient with CRC developed secondary metastases;one patient with HCC developed a local recurrence.Recurrence-free patients showed a signifi cantly higher cytolytic activity of PBMC against matched tumor cells 12 months after RFA treatment.Interestingly,patients with malignant recurrence showed a decreased cytolytic activity.Conclusion:RFA seems to overcome immune-tolerance toward tumor antigens and/or presents new tumor antigens.Patients seem to benefi t from a prolonged increase in cytolytic activity.The immune-modulatory effects of RFA need further investigations in multimodality anticancer therapies.展开更多
Arylamine N-acetyltransferase(NAT; E.C. 2.3.1.5) enzymes are responsible for the biotransformation of several arylamine and hydrazine drugs by acetylation. In this process, the acetyl group transferred to the acceptor...Arylamine N-acetyltransferase(NAT; E.C. 2.3.1.5) enzymes are responsible for the biotransformation of several arylamine and hydrazine drugs by acetylation. In this process, the acetyl group transferred to the acceptor substrate produces NAT deacetylation and, in consequence, it is susceptible of degradation. Sirtuins are protein deacetylases, dependent on nicotine adenine dinucleotide,which perform post-translational modifications on cytosolic proteins. To explore possible sirtuin participation in the enzymatic activity of arylamine NATs, the expression levels of NAT1, NAT2,SIRT1 and SIRT6 in peripheral blood mononuclear cells(PBMC) from healthy subjects were examined by flow cytometry and Western blot. The in situ activity of the sirtuins on NAT enzymatic activity was analyzed by HPLC, in the presence or absence of an agonist(resveratrol) and inhibitor(nicotinamide) of sirtuins. We detected a higher percentage of positive cells for NAT2 in comparison with NAT1, and higher numbers of SIRT1t cells compared to SIRT6 in lymphocytes. In situ NAT2 activity in the presence of NAM inhibitors was higher than in the presence of its substrate, but not in the presence ofresveratrol. In contrast, the activity of NAT1 was not affected by sirtuins. These results showed that NAT2 activity might be modified by sirtuins.展开更多
基金This study was supported by grants from the Key Project of the Chinese Ministry of Science and Technology(2017ZX102022022)National Key Research and Development Program of China(2021YFC2301801).
文摘Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnostic value for hepatitis B virus-associated hepatocellular carcinoma(HBV-HCC). Methods: We enrolled 197 patients, 111 with HBV-HCC, 51 with chronic hepatitis B(CHB), and 35 healthy controls(HCs). The methylation status of GLDC promoter in peripheral mononuclear cells(PBMCs) was identified by methylation specific polymerase chain reaction(MSP). The mRNA expression was examined using real-time quantitative polymerase chain reaction(q PCR). Results: The methylation frequency of the GLDC promoter was significantly lower in HBV-HCC patients(27.0%) compared to that in CHB patients(68.6%) and HCs(74.3%)( P < 0.001). The methylated group had lower alanine aminotransferase level( P = 0.035) and lower rates of tumor node metastasis(TNM) Ⅲ/Ⅳ( P = 0.043) and T3/T4( P = 0.026). TNM stage was identified to be an independent factor for GLDC promoter methylation. GLDC mRNA levels in CHB patients and HCs were significantly lower than those in HBV-HCC patients( P = 0.022 and P < 0.001, respectively). GLDC mRNA levels were significantly higher in HBV-HCC patients with unmethylated GLDC promoters than those with methylated GLDC promoters( P = 0.003). The diagnostic accuracy of alpha-fetoprotein(AFP) combined with GLDC promoter methylation for HBV-HCC was improved compared with that of AFP alone(AUC: 0.782 vs. 0.630, P < 0.001). In addition, GLDC promoter methylation was an independent predictor for overall survival of HBV-HCC patients( P = 0.038). Conclusions: The methylation frequency of GLDC promoter was lower in PBMCs from HBV-HCC patients than that from patients with CHB and HCs. The combination of AFP and GLDC promoter hypomethylation significantly improved the diagnostic accuracy of HBV-HCC.
文摘Objective:To compare the effect of human chorionic gonadotropin(hCG)-producing peripheral blood mononuclear cells(PBMCs)and PBMCs activated by hCG in vitro and expressions of related immune genes in mouse implantation.Methods:hCG-producing PBMCs(transfected PBMC)and PBMCs activated by hCG in vitro were introduced into isolated mouse endometrial cells,while cell cultures were divided into four groups:the control,PBMC,transfected,and activated PBMC groups.The expression of studied genes(IL-1β,IL-6,Lif,and Vegf)was evaluated and blastocyst attachment on the cocultured cells(isolated endometrial cells and PBMC cells)was monitored in all four groups.Results:Data showed that expression decreased in the PBMC group compared to the treated PBMC(transfected and activated PBMCs)and increased in transfected PBMC compared to the activated PBMC.Attachment and migration of blastocysts were dramatically enhanced in the transfected PBMC group compared to the activated PBMC group(P<0.05).Conclusions:Use of hCG-producing PBMCs(transfected PBMC)has more influence on endometrial receptivity.
基金Supported by a grant from the Damp-Foundation(2016-04) to Schaffler H and Rohde S
文摘AIM To investigate disease-specific gene expression profiles of peripheral blood mononuclear cells(PBMCs) from Crohn's disease(CD) patients in clinical remission.METHODS Patients with CD in clinical remission or with very low disease activity according to the Crohn's disease activity index were genotyped regarding nucleotidebinding oligomerization domain 2(NOD2),and PBMCs from wild-type(WT)-NOD2 patients,patients with homozygous or heterozygous NOD2 mutations and healthy donors were isolated for further analysis.The cells were cultured with vitamin D,peptidoglycan(PGN) and lipopolysaccharide(LPS) for defined periods of time before RNA was isolated and subjected to microarray analysis using Clariom S assays and quantitative realtime PCR.NOD2-and disease-specific gene expression profiles were evaluated with repeated measure ANOVA by a general linear model.RESULTS Employing microarray assays,a total of 267 genes were identified that were significantly up-or downregulated in PBMCs of WT-NOD2 patients,compared to healthy donors after challenge with vitamin D and/or a combination of LPS and PGN(P < 0.05;threshold:≥ 2-fold change).For further analysis by real-time PCR,genes with known impact on inflammation and immunity were selected that fulfilled predefined expression criteria.In a larger cohort of patients and controls,a disease-associated expression pattern,with higher transcript levels in vitamin D-treated PBMCs from patients,was observed for three of these genes,CLEC5 A(P < 0.030),lysozyme(LYZ;P < 0.047) and TREM1(P < 0.023).Six genes were found to be expressed in a NOD2-dependent manner(CD101,P < 0.002;CLEC5 A,P < 0.020;CXCL5,P < 0.009;IL-24,P < 0.044;ITGB2,P < 0.041;LYZ,P < 0.042).Interestingly,the highest transcript levels were observed in patients with heterozygous NOD2 mutations.CONCLUSION Our data identify CLEC5 A and LYZ as CD-and NOD2-associated genes of PBMCs and encourage further studies on their pathomechanistic roles.
基金supported by grants from FAPESP(Fundacao de AmparoàPesquisa de Sao Paulo,Grant no 2016/01302-9 and 2014/27129-6)CAPES(Coordenacao de Aperfeicoamento de Pessoal de Nível Superior)88887.463672/2019-00。
文摘Antipsychotics may prolong or retain telomere length,affect mitochondrial function,and then affect the metabolism of nerve cells.To validate the hypothesis that antipsychotics can prolong telomere length after oxidative stress injury,leukocytes from healthy volunteers were extracted using Ficoll-Histopaque density gradient.The mononuclear cells layer was resuspended in cell culture medium.Oxidative stress was induced with hydrogen peroxide in cultured leukocytes.Four days later,leukocytes were treated with aripiprazole,haloperidol or clozapine for 7 days.Real-time PCR revealed that treatments with aripiprazole and haloperidol increased the telomere length by 23%and 20%in peripheral blood mononuclear cells after acute oxidative stress injury.These results suggest that haloperidol and aripiprazole can reduce the damage to telomeres induced by oxidative stress.The experiment procedure was approved by the Ethics Committee of Faculty of Medicine of the University of Sao Paulo(FMUSP/CAAE approval No.52622616.8.0000.0065).
文摘To explore changes of toll-like receptor (TLR) 2,4 in peripheral blood mononuclear cells (PBMC) in acute abdomen patients with systemic inflammatory response syndrome (SIRS) and their significance.Methods A clinical study was done on 103 patients of which 65 were with SIRS.The mRNA expression of TLR2,4 were detected by RT-PCR;the expression of TNF-α and IL-6 were observed by ELISA;the correlation between TLR2,4 mRNA,the level of TNF-α and IL-6,and the clinical course was evaluated.Results TLR2 mRNA ,TNF-α and IL-6 were upregulated markedly on the first day of hospitalization,then decreased gradually;TLR2 mRNA maintained on high level till the 5th day.The expression of TLR2,4 mRNA was positive correlated with the level of TNF-α and IL-6,and the length of stay.TLR2,4 mRNA expression increased in patients with multiple organ failure.Conclusion In actue abdomen patients with SIRS,the expression of TLR2,4 of PBMC increased markedly,indicating its improtant role in the pathogenesis of SIRS.4 refs,2 figs,2 tabs.
基金National Key R&D Program of China(no.2022YFC2704501,2022YFC2704503 and 2022YFC2704505)National Natural Science Foundation of China(no.81830045 and 82071652)+1 种基金Sino-German Center for Research Promotion’s Rapid Response Funding Call for Bilateral Collaborative Proposals Between China and Germany in COVID-19 Related Research(grant/award no.C-0032)General Program of Guangdong Province Natural Science Foundation(no.2021A1515011039 and 2020A1515010273)。
文摘Objective:This study aimed to investigate the immune response of a pregnant woman who recovered from the coronavirus disease 2019(COVID_(R)S)by using single-cell transcriptomic profiling of peripheral blood mononuclear cells(PBMCs)and to analyze the properties of different immune cell subsets.Methods:PBMCs were collected from the COVID_(R)S patient at 28 weeks of gestation,before a cesarean section.The PBMCs were then analyzed using single-cell RNA sequencing.The transcriptional profiles of myeloid,T,and natural killer(NK)cell subsets were systematically analyzed and compared with those of healthy pregnant controls from a published single-cell RNA sequencing data set.Results:We identified major cell types such as T cells,B cells,NK cells,and myeloid cells in the PBMCs of our COVID_(R)S patient.The increase of myeloid and B cells and decrease of T cells and NK cells in the PBMCs in this patient were quite distinct compared with that in the control subjects.After reclustering and Augur analysis,we found that CD16 monocytes and mucosal-associated invariant T(MAIT)cells were mostly affected within different myeloid,T,and NK cell subtypes in our COVID_(R)S patient.The proportion of CD16 monocytes in the total myeloid population was increased,and the frequency of MAIT cells in the total T and NK cells was significantly decreased in the COVID-RS patient.We also observed significant enrichment of gene sets related to antigen processing and presentation,T-cell activation,T-cell differentiation,and tumor necrosis factor superfamily cytokine production in CD16 monocytes,and enrichment of gene sets related to antigen processing and presentation,response to type II interferon,and response to virus in MAIT cells.Conclusion:Our study provides a single-cell resolution atlas of the immune gene expression patterns in PBMCs from a COVID_(R)S patient.Our findings suggest that CD16-positive monocytes and MAIT cells likely play crucial roles in the maternal immune response against severe acute respiratory syndrome coronavirus 2 infection.These results contribute to a better understanding of the maternal immune response to severe acute respiratory syndrome coronavirus 2 infection and may have implications for the development of effective treatments and preventive strategies for the coronavirus disease 2019 in pregnant women.
基金supported by National Natural Science Foundation Key Program,China(No.81930023)Natural Science Foundation Major International(Regional)Joint Research Project,China(No.81720108009)+2 种基金Chongqing Outstanding Scientists Project(2019),Chongqing Key Laboratory of Ophthalmology,China(No.CSTC,2008CA5003)Chongqing Science&Technology Platform and Base Construction Program,China(No.cstc2014ptsy10002)the Chongqing Chief Medical Scientist Project,China(2018).
文摘Vogt-Koyanagi-Harada disease(VKH)is a rare autoimmune disease characterized by diffuse and bilateral uveitis,alopecia,tinnitus,hearing loss,vitiligo and headache.The transcriptional expression pattern of peripheral blood mononuclear cells(PBMC)in VKH remains largely unknown.In this study,mRNA sequencing was conducted in PBMC from VKH patients with active uveitis before treatment(n=7),the same patients after prednisone combined with cyclosporine treatment(n=7)and healthy control subjects strictly matched with gender and age(n=7).We found 118 differentially expressed genes(DEGs)between VKH patients and healthy control subjects,and 21 DEGs between VKH patients before and after treatment.TRIB1 was selected as a potential biomarker to monitor the development of VKH according to the mRNA sequencing.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were performed to predict the possible biological functions and signaling pathways of DEGs.Neutrophil degranulation,peptidase regulator activity,secretory granule membrane,cellular response to peptide,growth factor binding and cell projection membrane were enriched as GO annotations of DEGs.Arachidonic acid metabolism and mitogen-activated protein kinase(MAPK)signaling pathway were potential signaling pathways involved in pathogenesis and drug response of VKH.A protein–protein interaction(PPI)network was constructed by STRING,and colony stimulating factor 1 receptor(CSF1R)was identified as the hubgene of all DEGs by Cytoscape.The cell type presumed to contribute to the aberrant expression of DEGs was analyzed with the use of publicly available single-cell sequencing data of PBMC from a healthy donor and single-cell sequencing dataset of monocytes from VKH patients.Our findings may help to decipher the underlying cellular and molecular pathogenesis of VKH and may lead novel therapeutic applications.
基金This work was supported by the National Research Program of China(2017YFA0505803 and 2017YFC0908100)the State Key Project for Infectious Diseases(2018ZX10732202-001)+2 种基金National Natural Science Foundation of China(81790633,81672860,61922047,81422032,and 81902412)National Natural Science Foundation of Shanghai,China(17ZR143800)National Science Foundation for Distinguished Young Scholars of China(81702298).
文摘Intrahepatic cholangiocarcinoma(ICC)is the second most common liver cancer.Chemotherapy remains the main therapeutic strategy for advanced ICC patients,but chemosensitivity varies individually.Here,we applied cytometry by time-of-flight(CyTOF)to establish the immune profile of peripheral blood mononuclear cells(PBMCs)on the single-cell level at indicated time points before,during,and after chemotherapy.Multiplex immunofluorescence staining was applied to examine the spatial distribution of certain immune clusters.Tissue microarrays(TMAs)were used for prognostic evaluation.A total of 20 ICC patients treated with gemcitabine(GEM)were enrolled in our study,including eight cases with good response(R)and 12 cases with non-response(NR).Tremendous changes in PBMC composition,including an increased level of CD4/CD8 double-positive T cells(DPT),were observed after chemotherapy.Patients with higher level of CD4^(+)CD45RO^(+)CXCR3^(+)T cells before treatment had a favorable response to chemotherapy.Our study identified a positive correlation between the percentage of T cell subpopulations and clinical response after chemotherapy,which suggests that it is practical to predict the potential response before treatment by evaluating the proportions of the cell population in PBMCs.
基金Supported by MAHSA University Grant,No.RP158-05/19.
文摘In recent years,several studies have reported positive outcomes of cell-based therapies despite insufficient engraftment of transplanted cells.These findings have created a huge interest in the regenerative potential of paracrine factors released from transplanted stem or progenitor cells.Interestingly,this notion has also led scientists to question the role of proteins in the secretome produced by cells,tissues or organisms under certain conditions or at a particular time of regenerative therapy.Further studies have revealed that the secretomes derived from different cell types contain paracrine factors that could help to prevent apoptosis and induce proliferation of cells residing within the tissues of affected organs.This could also facilitate the migration of immune,progenitor and stem cells within the body to the site of inflammation.Of these different paracrine factors present within the secretome,researchers have given proper consideration to stromal cell-derived factor-1(SDF1)that plays a vital role in tissue-specific migration of the cells needed for regeneration.Recently researchers recognized that SDF1 could facilitate site-specific migration of cells by regulating SDF1-CXCR4 and/or HMGB1-SDF1-CXCR4 pathways which is vital for tissue regeneration.Hence in this study,we have attempted to describe the role of different types of cells within the body in facilitating regeneration while emphasizing the HMGB1-SDF1-CXCR4 pathway that orchestrates the migration of cells to the site where regeneration is needed.
基金supported by the National Natural Science Foundation of China (No.60178034)Doctor Thesis Founda-tion of Xi an Jiaotong University (No. DFXJTU2002-2) .
文摘Objective To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem cells (PBSCs), and furthermore, to understand the possible causes of this response. Methods We used MTT assay to detect the survival rate of PBMCs, CBMCs and PBSCs after treated by ALA-PDT under the optimum experiment conditions with U937 as control; Annexin V-FITC/PI was used to detect the pattern of cell death induced by ALA-PDT. By using flow cytometry, we detected intracellular PpIX fluorescence intensity. Results After ALA-PDT treatment the survival rate of PBMCs had no significant change; however in PBSCs and CBMCs, the survival rate reduced to 70%, and the survival rate of leukemia cell U937 was the lowest, about 30%. After incubation with ALA, except for PBMCs, intracellular PpIX fluorescence intensity of the other two kinds of normal haemocytes and U937 increased obviously. These results combined with the flow cytometry suggested that the main pattern of cell death here was apoptosis. Conclusion Under the optimum experiment conditions, ALA-PDT has a slight effect on normal haemocytes but excellent depletions of leukemia cells. Therefore, it can effectively purify autologous bone marrow or stem cell grafts.
基金Supported by the Natural Science Foundation of Shandong Province,No.2014ZRB01466
文摘AIM To explore the exact interaction between Notch and transforming growth factor(TGF)-β signaling in liver fibrosis. METHODS We established a rat model of liver fibrosis induced by concanavalin A. Peripheral blood mononuclear cells(PBMCs) were isolated from the modeled rats, and cultured with γ-secretase inhibitor DAPT and TGF-β inhibitor for 24 h. The m RNA levels of Notch and TGF-β signaling were detected by quantitative real-time polymerase chain reaction. Expression of Notch and TGF-β proteins was analyzed by western blotting.RESULTS Compared to control rats, Notch and TGF-β signaling was activated in PBMCs of model rats. Administration of DAPT and TGF-β inhibitor suppressed Notch and TGF-β signal transducer in PBMCs of model rats. DAPT reduced m RNA and protein expression of TGF-β signaling, such as TGF-β1 and Smad3. TGF-β inhibitor also downregulated Notch1, Hes1 and Hes5, and m RNA and protein expression of the Notch signaling pathway.CONCLUSION Notch and TGF-β signaling play a role in liver fibrosis. TGF-β signaling upregulates Notch signaling, which promotes TGF-β signaling.
基金Supported by Local Fund from Iran University of Medical Sciences
文摘AIM:To investigate the presence of mixed infection and discrepancy between hepatitis C virus(HCV) genotypes in plasma,peripheral blood mononuclear cells(PBMCs),and liver biopsy specimens.METHODS:From September 2008 up to April 2009,133 patients with chronic hepatitis C referred to Firouzgar Hospital for initiation of an antiviral therapy were recruited in the study.Five milliliters of peripheral blood was collected from each patient and liver biopsy was performed in those who gave consent or had indications.HCV genotyping was done using INNO-LiPATM HCV in serum,PBMCs,and liver biopsy specimens and then conf irmed by sequencing of 5'-UTR fragments.RESULTS:The mean age of patients was 30.3 ± 17.1 years.Multiple transfusion was seen in 124(93.2%) of patients.Multiple HCV genotypes were found in 3(2.3%) of 133 plasma samples,9(6.8%) of 133 PBMC samples,and 8(18.2%) of 44 liver biopsy specimens.It is notable that the different genotypes found in PBMCs were not the same as those found in plasma and liver biopsy specimens.CONCLUSION:Our study shows that a signif icant proportion of patients with chronic hepatitis C are affected by multiple HCV genotypes which may not be detectable only in serum of patients.
文摘Glaucoma is a neurodegenerative disease in which optic nerve damage and visual field defects occur.It is a leading cause of irreversible blindness.Its pathogenesis is largely unknown although several risk factors have been identified,with an increase in intraocular pressure being the main one.Lowering of intraocular pressure is the only treatment available.Open-angle glaucoma is the most common form of the condition,accounting for~90%of all cases of glaucoma,with primary open-angle glaucoma and exfoliation glaucoma being the most frequent types.There are strong indications that microRNAs play important roles in the pathogenesis of primary open-angle glaucoma.Most of the recent studies reviewed had performed microRNA profiling in aqueous humor from glaucoma patients compared to controls who were chiefly cataract patients.A very large number of microRNAs were dysregulated but with limited overlap between individual studies.MiRNAs in aqueous humor that could be possible targets for therapeutic intervention are miR-143-3p,miR-125b-5p,and miR-1260b.No ove rlap of findings occurred within the dysregulated miRNAs for blood plasma,blood serum,peripheral blood mononuclear cells,and tears of primary open-angle glaucoma patients.Seve ral impo rtant limitations were identified in these studies.Further studies are warranted of mic roRNA expression in aqueous humor and blood samples of primary open-angle glaucoma patients in the early stages of the disease so that validated biomarkers can be identified and treatment initiated.In addition,whether modifying the levels of specific microRNAs in aqueous humor or tears has a beneficial effect on intraocular pressure and ophthalmic examination of the eyes should be investigated using suitable animal models of glaucoma.
文摘Objective To evaluate the methylation status at CpG site -55 in the interferon-gamma (IFN-γ) gene promoter and its effect on IFN-γ expression in chronic hepatitis B. Method The authors recruited 30 patients with HBeAg-positive chronic hepatitis B (CHB), 30 HBeAg-negative CHB patients, and 30 healthy blood donors. Pyrosequencing was used to determine the methylation status at CpG site -55 in the IFN-γ gene promoter following bisulfite treatment of DNA in peripheral blood mononuclear cells (PBMCs). The expression of IFN-γ was analyzed by real-time RT-PCR and ELISA. HBV DNA in PBMCs was detected by nested PCR. Results The methylation level at CpG site -55 in the IFN-γ gene promoter was significantly increased, resulting in subsequent down-regulation of the expression of this cytokine in CHB. The methylation level at CpG site -55 was significantly higher in HBeAg-positive patients than in HBeAg-negative ones (P<0.01) and was also significantly higher in PBMCs from HBV DNA-positive patients than from HBV DNA-negative ones (P<0.01); the methylation level at CpG site -55 was positively correlated with the amount of HBV DNA in serum (P<0.01). Conclusion IFN-γ gene expression appears to be regulated by methylation of the IFN-γ gene promoter in CHB; the methylation level at CpG site -55 is associated with HBV infection.
文摘In order to increase the positive detection rate of HCV RNA in the patients with chronic hepatitis C , RT PCR was used to synchronously detect HCV RNA in the plasma and peripheral blood mononuclear cells of 583 CHC patients with a continuously elevated level of ALT for more than one year. The results showed that the positive detection rate of HCV RNA in the plasma of the CHC patients was 19.2 %, while 24.5 % in PBMC. It was demonstrated that the positive detection rate for HCV RNA in PBMC was obviously higher than that detected in plasma. To synchronously detect HCV RNA in PBMC by using RT PCR can increase the positive detection rate of HCV RNA in the CHC patients.
基金Data and materials availability:Processed and raw data can be downloaded from NCBI GEO(#GSE122682,and#GSE153421).
文摘Although widely applied in treating hematopoietic malignancies,transplantation of hematopoietic stem/progenitor cells(HSPCs)is impeded by HSPC shortage.Whether circulating HSPCs(cHSPCs)in steady-state blood could be used as an alternative source remains largely elusive.Here we develop a three-dimensional culture system(3DCS)including arginine,glycine,aspartate,and a series of factors.Fourteen-day culture of peripheral blood mononuclear cells(PBMNCs)in 3DCS led to 125-and 70-fold increase of the frequency and number of CD34+cells.Further,3DCS-expanded cHSPCs exhibited the similar reconstitution rate com-pared to CD34+HSPCs in bone marrow.Mechanistically,3DCS fabricated an immunomodulatory niche,secreting cytokines as TNF to support cHSPC survival and proliferation.Finally,3DCS could also promote the expansion of cHSPCs in patients who failed in HSPC mobilization.Our 3DCS successfully expands rare cHSPCs,providing an alternative source for the HSPC therapy,particularly for the patients/donors who have failed in HSPC mobilization.
文摘BACKGROUND Occult hepatitis C infection(OCI)is characterized by the presence of hepatitis C virus(HCV)RNA in the liver,peripheral blood mononuclear cells(PBMC)and/or ultracentrifuged serum in the absence of detectable HCV-RNA in serum.OCI has been described in several categories of populations including hemodialysis patients,patients with a sustained virological response,immunocompromised individuals,patients with abnormal hepatic function,and apparently healthy subjects.AIM To highlight the global prevalence of OCI.METHODS We performed a systematic and comprehensive literature search in the following 4 electronic databases PubMed,EMBASE,Global Index Medicus,and Web of Science up to 6th May 2021 to retrieve relevant studies published in the field.Included studies were unrestricted population categories with known RNA status in serum,PBMC,liver tissue and/or ultracentrifuged serum.Data were extracted independently by each author and the Hoy et al tool was used to assess the quality of the included studies.We used the random-effect meta-analysis model to estimate the proportions of OCI and their 95%confidence intervals(95%CI).The Cochran's Q-test and the I2 test statistics were used to assess heterogeneity between studies.Funnel plot and Egger test were used to examine publication bias.R software version 4.1.0 was used for all analyses.RESULTS The electronic search resulted in 3950 articles.We obtained 102 prevalence data from 85 included studies.The pooled prevalence of seronegative OCI was estimated to be 9.61%(95%CI:6.84-12.73)with substantial heterogeneity[I^(2)=94.7%(95%CI:93.8%-95.4%),P<0.0001].Seropositive OCI prevalence was estimated to be 13.39%(95%CI:7.85-19.99)with substantial heterogeneity[I^(2)=93.0%(90.8%-94.7%)].Higher seronegative OCI prevalence was found in Southern Europe and Northern Africa,and in patients with abnormal liver function,hematological disorders,and kidney diseases.Higher seropositive OCI prevalence was found in Southern Europe,Northern America,and Northern Africa.CONCLUSION In conclusion,in the present study,it appears that the burden of OCI is high and variable across the different regions and population categories.Further studies on OCI are needed to assess the transmissibility,clinical significance,long-term outcome,and need for treatment.
基金supported by Conselho Nacional de Desenvolviment Coientificoe Tecnologico(CNPq310100/2017-8,403201/2020-9,and INCT 465539/2014-9)+3 种基金Fundagaode Amparoa Pesquisado Estado deSao Paulo(FAPESP2013/16349-2 and 2014/02438-6)the National Institutes of Health(NIHAl163019).M.C.P and R.G.were funded by CNPq(380849/2020-8).
文摘The severe acute respiratory syndrome coronavirus 2(SARS-CovV-2)infection is associated with a hyperinflammatory state and lymphocytopenia,a hallmark that appears as both signature and prognosis of disease severity outcome.Although cytokine storm and a sustained inflammatory state are commonly associated with immune cell depletion,it is still unclear whether direct SARS-Cov-2 infection of immune cells could also play a role in this scenario by harboring viral replication.We found that monocytes,as well as both B and T lymphocytes,were susceptible to SARS-Cov-2 infection in vitro,accumulating double-stranded RNA consistent with viral RNA replication and ultimately leading to expressive T cell apoptosis.In addition,flow cytometry and immunofluorescence analysis revealed that SARS-Cov-2 was frequently detected in monocytes and B lymphocytes from coronavirus disease 2019(CoVID-19)patients.The rates of SARS-Cov-2-infected monocytes in peripheral blood mononuclear cells from CoVID-19 patients increased over time from symptom onset,with SARS-CoV-2-positive monocytes,B cells,and CD4+T lymphocytes also detected in postmortem lung tissue.These results indicated that SARS-CoV-2 infection of blood-circulating leukocytes in covID-19 patients might have important implications for disease pathogenesis and progression,immune dysfunction,and virus spread within the host.
基金This project was supported by the Hans-Loewel foundation Bamberg,Germany.
文摘Aim:Used as a palliative therapy for unresectable liver cancer,radiofrequency ablation(RFA)is associated with the induction of immunological responses.Here,we show strong evidence of tumor-specifi c peripheral blood mononuclear cells(PBMCs)12 months after RFA.Methods:Three patients with colorectal cancer(CRC)metastases to the liver and two patients with primary hepatocellular carcinoma(HCC)were enrolled in this study.PBMC,isolated 12 months after RFA,were stimulated with normal and tumor tissue lysate.Interferon gamma secretion was evaluated by fl ow cytometry and indirectly,by luciferase assay for adenylate kinase activity in PBMC-stimulated lysates of target cells.Baseline data were detected before RFA and 4 weeks after treatment.Results:Two CRC patients and one HCC patient had recurrence-free survival.One patient with CRC developed secondary metastases;one patient with HCC developed a local recurrence.Recurrence-free patients showed a signifi cantly higher cytolytic activity of PBMC against matched tumor cells 12 months after RFA treatment.Interestingly,patients with malignant recurrence showed a decreased cytolytic activity.Conclusion:RFA seems to overcome immune-tolerance toward tumor antigens and/or presents new tumor antigens.Patients seem to benefi t from a prolonged increase in cytolytic activity.The immune-modulatory effects of RFA need further investigations in multimodality anticancer therapies.
基金supported by grants 255781 and 248950 (to Portales-Perez Diana Patricia) from CONACYT, MexicoTuriján-Espinoza Eneida was the recipient of a scholarship (592537) from CONACYT, Mexico
文摘Arylamine N-acetyltransferase(NAT; E.C. 2.3.1.5) enzymes are responsible for the biotransformation of several arylamine and hydrazine drugs by acetylation. In this process, the acetyl group transferred to the acceptor substrate produces NAT deacetylation and, in consequence, it is susceptible of degradation. Sirtuins are protein deacetylases, dependent on nicotine adenine dinucleotide,which perform post-translational modifications on cytosolic proteins. To explore possible sirtuin participation in the enzymatic activity of arylamine NATs, the expression levels of NAT1, NAT2,SIRT1 and SIRT6 in peripheral blood mononuclear cells(PBMC) from healthy subjects were examined by flow cytometry and Western blot. The in situ activity of the sirtuins on NAT enzymatic activity was analyzed by HPLC, in the presence or absence of an agonist(resveratrol) and inhibitor(nicotinamide) of sirtuins. We detected a higher percentage of positive cells for NAT2 in comparison with NAT1, and higher numbers of SIRT1t cells compared to SIRT6 in lymphocytes. In situ NAT2 activity in the presence of NAM inhibitors was higher than in the presence of its substrate, but not in the presence ofresveratrol. In contrast, the activity of NAT1 was not affected by sirtuins. These results showed that NAT2 activity might be modified by sirtuins.
