Comparison of the conventional tube and erythrocyte-magnetized technology in titration of red blood cell alloantibodies

BACKGROUND Erythrocyte alloantibodies are mainly produced after immune stimulation,such as blood transfusion,pregnancy,and transplantation,and are the leading causes of severe hemolytic transfusion reactions and difficulty in blood grouping and matching.Therefore,antibody screening is critical to prevent and improve red cell alloantibodies.Routine tube assay is the primary detection method of antibody screening.Recently,erythrocyte-magnetized technology(EMT)has been increasingly used in clinical practice.This study intends to probe the application and efficacy of the conventional tube and EMT in red blood cell alloantibody titration to provide a reference for clinical blood transfusion.AIM To investigate the application value of conventional tube and EMT in red blood cell alloantibody titration and enhance the safety of blood transfusion practice.METHODS A total of 1298 blood samples were harvested from blood donors at the Department of Blood Transfusion of our hospital from March 2021 to December 2022.A 5 mL blood sample was collected in tubing,which was then cut,and the whole blood was put into a test tube for centrifugation to separate the serum.Different red blood cell blood group antibody titers were simultaneously detected using the tube polybrene test,tube antiglobulin test(AGT),and EMT screening irregular antibody methods to determine the best test method.RESULTS Simultaneous detection was performed through the tube polybrene test,tube AGT and EMT screening irregular antibodies.It was discovered that the EMT screening irregular antibody method could detect all immunoglobulin G(IgG)and immunoglobulin M(IgM)irregular antibodies,and the results of manual tube AGT were satisfactory,but the operation time was lengthy,and the equipment had a large footprint.The EMT screening irregular antibody assay was also conducted to determine its activity against type O Rh(D)red blood cells,and the outcomes were satisfactory.Furthermore,compared to the conventional tube method,the EMT screening irregular antibody method was more cost-effective and had significantly higher detection efficiency.CONCLUSION With a higher detection rate,the EMT screening irregular antibody method can detect both IgG and IgM irregular antibodies faster and more effectively than the conventional tube method.

Significant increase in HBV, HCV, HIV and syphilis infections among blood donors in West Bengal, Eastern India 2004-2005: Exploratory screening reveals high frequency of occult HBV infection

AIM: To evaluate the prevalence of markers of hepatitis B virus (HBV) and hepatitis C virus (HCV) and human immunodeficiency virus (HIV) among blood donors in Kolkata, Eastern India for two consecutive years and to conduct a pilot study to explore the presence of HBV DNA among hepatitis B surface antigen (HBsAg) negative but anti-HBc positive blood donors. METHODS: Seroprevalence of HBsAg, anti-HCV and anti-HIV was studied among 113 051 and 106 695 voluntary blood donors screened in 2004 and 2005, respectively. Moreover, a pilot study on 1027 HBsAg negative donors was carried out for evaluating the presence of HBV DNA by PCR on HBsAg negative/anti- HBc positive donors. RESULTS: A statistically significant increase in the prevalence of HBV (1448 vs 1768, P < 0.001), HIV (262 vs 374, P < 0.001), HCV (314 vs 372, P = 0.003) and syphilis (772 vs 853, P = 0.001) infections was noted among blood donors of Kolkata West Bengal in 2005 as compared to 2004. Moreover, the exploratory study on 1027 HBsAg negative donors revealed that 188 (18.3%)of them were anti-HBc positive out of which 21% were positive for HBV DNA. CONCLUSION: The findings of this study underscore the significantly increasing endemicity of hepatitis viruses, syphilis and HIV among the voluntary blood donors of our community. The pilot study indicates a high rate of prevalence of HBV DNA among HBsAg negative/anti-HBc positive donors and thus emphasizes the need for a more sensitive and stringent screening algorithm for blood donations.

Role of noncoding RNAs in liver fibrosis

Liver fibrosis is a wound-healing response following chronic liver injury caused by hepatitis virus infection,obesity,or excessive alcohol.It is a dynamic and reversible process characterized by the activation of hepatic stellate cells and excess accumulation of extracellular matrix.Advanced fibrosis could lead to cirrhosis and even liver cancer,which has become a significant health burden worldwide.Many studies have revealed that noncoding RNAs(ncRNAs),including microRNAs,long noncoding RNAs and circular RNAs,are involved in the pathogenesis and development of liver fibrosis by regulating signaling pathways including transforming growth factor-βpathway,phosphatidylinositol 3-kinase/protein kinase B pathway,and Wnt/β-catenin pathway.NcRNAs in serum or exosomes have been reported to tentatively applied in the diagnosis and staging of liver fibrosis and combined with elastography to improve the accuracy of diagnosis.NcRNAs mimics,ncRNAs in mesenchymal stem cell-derived exosomes,and lipid nanoparticles-encapsulated ncRNAs have become promising therapeutic approaches for the treatment of liver fibrosis.In this review,we update the latest knowledge on ncRNAs in the pathogenesis and progression of liver fibrosis,and discuss the potentials and challenges to use these ncRNAs for diagnosis,staging and treatment of liver fibrosis.All these will help us to develop a comprehensive understanding of the role of ncRNAs in liver fibrosis.

Ferroptosis in liver diseases:Fundamental mechanism and clinical implications

This editorial discusses a recently published paper in the World Journal of Gastroenterology.Our research focuses on p53's regulatory mechanism for controlling ferroptosis,as well as the intricate connection between ferroptosis and liver diseases.Ferroptosis is a specific form of programmed cell death that is dependent on iron and displays unique features in terms of morphology,biology,and genetics,distinguishing it from other forms of cell death.Ferroptosis can affect the liver,which is a crucial organ responsible for iron storage and metabolism.Mounting evidence indicates a robust correlation between ferroptosis and the advancement of liver disorders.P53 has a dual effect on ferroptosis through various distinct signaling pathways.However,additional investigations are required to clarify the regulatory function of p53 metabolic targets in this complex association with ferroptosis.In the future,researchers should clarify the mechanisms by which ferroptosis and other forms of programmed cell death contribute to the progression of liver diseases.Identifying and controlling important regulatory factors associated with ferroptosis present a promising therapeutic strategy for liver disorders.

A CASE OF HEMOLYTIC DISEASE OF THE NEWBORN CAUSED BY ANTI-HRO AND ANTI-E

A Chinese woman of blood group B,D-and her husband of blood group AB,CCDeewere examined.The woman had not been transfused before.Their first two babiesdied.Anti-Hro and anti-e were found in the mother’s serum.During her third pregnancy,the titer of antibodies went up quickly,approximately one titer per month.After 36 weeksof pregnancy,the baby was delivered by Caesarean section.The cord blood Hb was 88g/L,his red blood cell count 2.7×1012/L,and total biIirubin 114.6 mol/L.The baby was ofblood group AB,and CDe-D-genotype.Exchangetransfusion was begun 2.5 hours afterbirth.O,ccDEE washed red cells together with group AB plasma were used.Two dayslater,7Oml washed O,ccDEE concentrated red cells were administered.The baby is aliveand in good health.

Photoinduced DNA Cleavage of Fullerols,Water-Soluble Polyhydroxylated [C_(60)] Fullerene Derivatives

The effects of polyhydroxylated [C60 ] fullerene derivatives fullerols on DNA was studied, using the piasmid pXJ41-neo DNA as the experimental model. The cleaved DNA products were detected by agarose gel electrophoresis. The results showed that fullerols could stimulate DNA cleavage in dose and irradiation dependent manners. 0.4 mmol/L fullerols together with 1.5 h exposure to a 500 W tungsten halogen lamp at a distance of 20 cm could convert most of plasmid DNA from the intact form into the nicked and linear forms. Scavengers of various reactive oxygen species （ROS） including sodium azide, mannitol and superoxide dismu- tase （SOD） could inhibit the photoinduced DNA cleavage of fuUerols. These data presented for the first time the photoinduced biological activities of fullerols, and implied a possible use of these fullerene derivatives as the candidates for novel photosensitizers in the biomedical therapy.

Serum folate status among blood donors in south-central China

Folate deficiency has been confirmed to be related to various diseases.Unfortunately,there are few reports on the folate status of Chinese adults.This study aims to evaluate the serum folate status of blood donors in south-central China.In this study,248 blood donors were included.The information on subjects was collected by a brief questionnaire concerning alcohol consumption habits,smoking habits,fruit and vegetable consumption and physical activity.The serum folate concentration was measured by electrochemiluminescence immunoassay.The geometric mean serum folate concentration was 13.4 nmoll-1(95%CI,12.7-14.1).The prevalence of serum folate concentrations below 6.8 nmoll-1 was 5.2%(95%CI,2.5-8.0).There were significant differences in serum folate concentrations with respect to sex(p-values<0.05),age(p-values<0.05),fruit and vegetable consumption(p-values<0.05),and alcohol consumption habits(p-values<0.05).The concentration of serum folate increased with age(p-values<0.05)and fruit and vegetable consumption(p-values<0.05).Individuals with an age of 30 years or younger were nearly 3.5 times as likely as those aged over 30 years to have an insufficient level of serum folate(OR=3.48;95%CI:1.01-11.99).An age of 30 years or younger was a risk factor for folate deficiency.Most blood donors had sufficient serum folate concentrations in south-central China.National surveys of folate status should be implemented in China.

Role of Exosomal Modulation of Macrophages in Liver Fibrosis

Exosomes are 60–120 nm diameter double-membrane lipid organelles discharged by cells.Various studies have shown that exosomes exert multiple functions in both physical and diseased processes,such as intercellular information exchange,immune response,and disease progression.Repeated chronic injury to the liver often leads to inflammation and liver fibrosis(LF),a disorder that,if unchecked,may progress to cirrhosis,liver failure,portal hypertension,and even hepatocellular carcinoma.As an essential component of host innate immunity against pathogen invasion,macrophages play an important role in modulating inflammation homeostasis by finely tuning the polarization process of macrophages into either M1 or M2 subtypes in response to different microenvironments.As a critical contributor to the inflammation process,macrophages also play a complex and instrumental function in the progression of LF.This review focuses on recent advancements in the role of macrophage-associated exosomes implicated in LF,including macrophage-released exosomes and macrophage-targeted exosomes.In addition,the progress made in exosomebased antifibrotic therapy by in vivo and in vitro studies is also highlighted.

REDH:A database of RNA editome in hematopoietic differentiation and malignancy

Background:The conversion of adenosine(A)to inosine(I)through deamination is the prevailing form of RNA editing,impacting numerous nuclear and cytoplasmic transcripts across various eukaryotic species.Millions of high-confidence RNA editing sites have been identified and integrated into various RNA databases,providing a convenient platform for the rapid identification of key drivers of cancer and potential therapeutic targets.However,the available database for integration of RNA editing in hematopoietic cells and hematopoietic malignancies is still lacking.Methods:We downloaded RNA sequencing(RNA-seq)data of 29 leukemia patients and 19 healthy donors from National Center for Biotechnology Information(NCBI)Gene Expression Omnibus(GEO)database,and RNA-seq data of 12 mouse hematopoietic cell populations obtained from our previous research were also used.We performed sequence alignment,identified RNA editing sites,and obtained characteristic editing sites related to normal hematopoietic development and abnormal editing sites associated with hematologic diseases.Results:We established a new database,"REDH",represents RNA editome in hematopoietic differentiation and malignancy.REDH is a curated database of associations between RNA editome and hematopoiesis.REDH integrates 30,796 editing sites from 12 murine adult hematopoietic cell populations and systematically characterizes more than 400,000 edited events in malignant hematopoietic samples from 48 cohorts(human).Through the Differentiation,Disease,Enrichment,and knowledge modules,each A-to-I editing site is systematically integrated,including its distribution throughout the genome,its clinical information(human sample),and functional editing sites under physiological and pathological conditions.Furthermore,REDH compares the similarities and differences of editing sites between different hematologic malignancies and healthy control.Conclusions:REDH is accessible at http://www.redhdatabase.com/.This user-friendly database would aid in understanding the mechanisms of RNA editing in hematopoietic differentiation and malignancies.It provides a set of data related to the maintenance of hematopoietic homeostasis and identifying potential therapeutic targets in malignancies.

Exploring the potential of micro-nano composite structures for COVID-19 vaccines and beyond

The coronavirus disease 2019(COVID-19)pandemic has emphasised the crucial role of vaccination in mitigating the spread of the disease.1 While several severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)vaccines have been authorised,most of them are administered through intramuscular injections.2 Although these vaccines effectively elicit systemic immune responses,they do not provide immediate protection at the respiratory tract,which is the primary site of viral infection.3,4 To overcome this limitation,researchers are actively investigating the potential of intranasal or nebulised vaccine candidates.

经冠状动脉自体骨髓间充质干细胞移植治疗小型猪急性心肌梗死

目的评价经冠状动脉移植体外分离培养自体骨髓间充质干细胞(MSCs)治疗急性心肌梗死(AMI)的可行性及疗效。方法通过结扎冠状动脉左前降支90min后恢复血流的方法建立AMI模型。采用密度梯度离心法分离骨髓单个核细胞、传代培养后,在AMI造模10～14d后将培养扩增的自体MSCs经冠状动脉植入梗死区,应用超声心动图观察移植前后心功能变化情况,冰冻切片荧光显微镜示踪4'G6-二脒-2-苯基吲哚(DAPI)标记的MSCs,Ⅷ因子免疫荧光染色测定新生血管数目。结果MSCs生长呈纺锤样,生长旺盛呈旋涡样;DAPI标记率为100%。AMI造模10～14d后MSCs移植数量平均为(4～6)×107个,移植术后3个月,MSCs治疗组与对照组比较,左心室射血分数(LVEF)显著升高[(54.65±3.39)%vs(43.98±4.21)%(P<0.01)],可见DAPI标记胞核为蓝色荧光的移植细胞,MSCs治疗组与对照组比较,新生血管数目显著升高[(13.28±4.39)vs(4.27±2.28)个G(P<0.01)]。结论通过体外培养扩增在短期内可以获得相当数量的生物性状稳定的MSCs,在AMI造模10～14d后进行经冠状动脉MSCs自体移植,促进左室功能的恢复。MSCs移植与目前广泛运用的介入治疗相结合,方法简便、经济、有效,可望改善AMI患者的预后,具有良好的临床应用前景。

Combining single-cell profiling and functional analysis explores the role of pseudogenes in human early embryonic development

More and more studies have demonstrated that pseudogenes possess coding ability,and the functions of their transcripts in the development of diseases have been partially revealed.However,the role of pseudogenes in maintenance of normal physiological states and life activities has long been neglected.Here,we identify pseudogenes that are dynamically expressed during human early embryogenesis,showing different expression patterns from that of adult tissues.We explore the expression correlation between pseudogenes and the parent genes,partly due to their shared gene regulatory elements or the potential regulation network between them.The essential role of three pseudogenes,PI4KAP1,TMED10P1,and FBXW4P1,in maintaining self-renewal of human embryonic stem cells is demonstrated.We further find that the three pseudogenes might perform their regulatory functions by binding to proteins or microRNAs.The pseudogene-related single-nucleotide polymorphisms are significantly associated with human congenital disease,further illustrating their importance during early embryonic development.Overall,this study is an excavation and exploration of functional pseudogenes during early human embryonic development,suggesting that pseudogenes are not only capable of being specifically activated in pathological states,but also play crucial roles in the maintenance of normal physiological states.

Dysregulated liver function in SARS-CoV-2 infection:Current understanding and perspectives

Since it was first reported in December 2019,severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection has spread rapidly around the world to cause the ongoing pandemic.Although the clinical manifestations of SARS-CoV-2 infection are predominantly in the respiratory system,liver enzyme abnormalities exist in around half of the cases,which indicate liver injury,and raise clinical concern.At present,there is no consensus whether the liver injury is directly caused by viral replication in the liver tissue or indirectly by the systemic inflammatory response.This review aims to summarize the clinical manifestations and to explore the underlying mechanisms of liver dysfunction in patients with SARSCoV-2 infection.

Rudimentary study on humanization of porcine red blood cells: Enzymatic removal of galactose-α1,3- galactose antigen from porcine red blood cell

The serological and biochemical characteriza-tion of porcine red blood cells (pRBCs) are similar to human red blood cells. Porcine erythrocytes are considered as an alternative source for human blood transfusion. But there exist galactose-?,3-galactose antigens (Gal?,3Gal?, 4GalNAcR, abbreviated 酖al antigen) on pRBCs, which can induce anti-aGal antibodies in human serum. The aGal epitopes are the major antigen responsible for hyperacute rejection in xenotransfusion. In this study, recombined soy-bean -galactosidase (rS?GalE) was used to remove the aGal antigens from pPRCs for humanization. The results showed that aGal antigen was cleared by rS?GalE and the structure and function of rS?GalE treated pRBC were normal.

Hybrid liposome-erythrocyte drug delivery system for tumor therapy with enhanced targeting and blood circulation

Liposome,a widely used drug delivery system(DDS),still shows several disadvantages such as dominant clearance by liver and poor target organ deposition.To overcome the drawbacks of liposomes,we developed a novel red blood cell(RBC)-liposome combined DDS to modulate the tumor accumulation and extend the blood circulation life of the existing liposomal DDS.Here,RBCs,an ideal natural carrier DDS,were utilized to carry liposomes and avoid them undergo the fast clearance in the blood.In this study,liposomes could either absorbed onto RBCs’surface or fuse with RBCs’membrane by merely altering the interaction time at 37℃,while the interaction between liposome and RBCs would not affect RBCs’characteristics.In the in vivo antitumor therapeutic efficacy study,1,2-dipalmitoyl-sn-glycero-3-phosphocholine(DPPC)liposomes attached onto RBCs’surfaces exhibited lung targeting effect(via RBC-hitchhiking approach)and reduced clearance in the liver,while DPPC liposomes fused with RBCs had prolong blood circulation up to 48 h and no enrichment in any organ.Furthermore,20 mol%of DPPC liposomes were replaced with pH-sensitive phospholipid 1,2-dioleoyl-Sn-glycero-3-phosphoethanolamine(DOPE)as it could respond to the low pH tumor microenvironment and then accumulate in the tumor.The DOPE attached/fusion RBCs showed partial enrichment in lung and about 5-8%tumor accumulation,which were significantly higher than(about 0.7%)the conventional liposomal DDS.Thus,RBC-liposome composite DDS is able to improve the liposomal tumor accumulation and blood circulation and shows the clinical application promises of using autologous RBCs for antitumor therapy.

Serosurvey for SARS-CoV-2 among blood donors in Wuhan,China from September to December 2019

The emerging of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)caused COVID-19 pandemic.The first case of COVID-19 was reported at early December in 2019 in Wuhan City,China.To examine specific antibodies against SARS-CoV-2 in biological samples before December 2019 would give clues when the epidemic of SARS-CoV-2 might start to circulate in populations.We obtained all 88,517 plasmas from 76,844 blood donors in Wuhan between 1 September and 31 December 2019.We first evaluated the pan-immunoglobin(pan-Ig)against SARS-CoV-2 in 43,850 samples from 32,484 blood donors with suitable sample quality and enough volume.Two hundred and sixty-four samples from 213 donors were pan-Ig reactive,then further tested IgG and IgM,and validated by neutralizing antibodies against SARS-CoV-2.Two hundred and thirteen samples(from 175 donors)were only pan-Ig reactive,8(from 4 donors)were pan-Ig and IgG reactive,and 43(from 34 donors)were pan-Ig and IgM reactive.Microneutralization assay showed all negative results.In addition,213 screened reactive donors were analyzed and did not show obviously temporal or regional tendency,but the distribution of age showed a difference compared with all tested donors.Then we reviewed SARS-CoV-2 antibody results from these donors who donated several times from September 2019 to June 2020,partly tested in a previous published study,no one was found a significant increase in S/CO of antibodies against SARS-CoV-2.Our findings showed no SARS-CoV-2-specific antibodies existing among blood donors in Wuhan,China before 2020,indicating no evidence of transmission of COVID-19 before December 2019 in Wuhan,China.

Oncolytic Virus Senecavirus A Inhibits Hepatocellular Carcinoma Proliferation and Growth by Inducing Cell Cycle Arrest and Apoptosis

Background and Aims:Hepatocellular carcinoma(HCC)isa highly aggressive tumor with limited treatment options andhigh mortality.Senecavirus A(SVA)has shown potential inselectively targeting tumors while sparing healthy tissues.This study aimed to investigate the effects of SVA on HCCcells in vitro and in vivo and to elucidate its mechanisms ofaction.Methods:The cell counting kit-8 assay and colonyformation assay were conducted to examine cell proliferation.Flow cytometry and nuclear staining were employed toanalyze cell cycle distribution and apoptosis occurrence.Asubcutaneous tumor xenograft HCC mouse model was createdin vivo using HepG2 cells,and Ki67 expression in thetumor tissues was assessed.The terminal deoxynucleotidyltransferase dUTP nick end labeling assay and hematoxylinand eosin staining were employed to evaluate HCC apoptosisand the toxicity of SVA on mouse organs.Results:In vitro,SVA effectively suppressed the growth of tumor cells by inducingapoptosis and cell cycle arrest.However,it did nothave a notable effect on normal hepatocytes(MIHA cells).In an in vivo setting,SVA effectively suppressed the growthof HCC in a mouse model.SVA treatment resulted in a significantdecrease in Ki67 expression and an increase in apoptosisof tumor cells.No notable histopathological alterationswere observed in the organs of mice during SVA administration.Conclusions:SVA inhibits the growth of HCC cells byinducing cell cycle arrest and apoptosis.It does not causeany noticeable toxicity to vital organs.

Internal m^(6)A and m^(7)G RNA modifications in hematopoietic system and acute myeloid leukemia

Epitranscriptomics focuses on the RNA-modification-mediated post-transcriptional regulation of gene expression.The past decade has witnessed tremendous progress in our understanding of the landscapes and biological functions of RNA modifications,as prompted by the emergence of potent analytical approaches.The hematopoietic system provides a lifelong supply of blood cells,and gene expression is tightly controlled during the differentiation of hematopoietic stem cells(HSCs).The dysregulation of gene expression during hematopoiesis may lead to severe disorders,including acute myeloid leukemia(AML).Emerging evidence supports the involvement of the mRNA modification system in normal hematopoiesis and AML pathogenesis,which has led to the development of small-molecule inhibitors that target N6-methyladenosine(m^(6)A)modification machinery as treatments.Here,we summarize the latest findings and our most up-to-date information on the roles of m^(6)A and N7-methylguanine in both physiological and pathological conditions in the hematopoietic system.Furthermore,we will discuss the therapeutic potential and limitations of cancer treatments targeting m^(6)A.

Human mesenchymal stromal cells enhance the immunomodulatory function of CD8＋CD28- regulatory T cells

One important aspect of mesenchymal stromal cells （MSCs）-mediated immunomodulation is the recruitment and induction of regulatory T （Treg） cells. However, we do not yet know whether MSCs have similar effects on the other subsets of Treg cells. Herein, we studied the effects of MSCs on CD8＋CD28- Treg cells and found that the MSCs could not only increase the proportion of CD8＋CD28- T cells, but also enhance CD8＋CD28-T cells＇ ability of hampering naive CD4＋ T-cell proliferation and activation, decreasing the production of IFN-γ by activated CD4＋ T cells and inducing the apoptosis of activated CD4＋ T cells. Mechanistically, the MSCs affected the functions of the CD8＋CD28- T cells partially through moderate upregulating the expression of IL-10 and FasL. The MSCs had no distinct effect on the shift from CD8＋CD28＋ T cells to CD8＋CD28- T cells, but did increase the proportion of CD8＋CD28- T cells by reducing their rate of apoptosis. In summary, this study shows that MSCs can enhance the regulatory function of CD8＋CD28- Treg cells, shedding new light on MSCs-mediated immune regulation.

Cloning of aminopeptidase N promoter and its activity in hematopoietic cell and different tumor cell lines

Aminopeptidase N (APN) promoter region was cloned and sequenced from peripheral blood mononuclear cells. The recombinant reporter construct containing the promoter and luciferase gene, designated pXPl-APNLuc, was introduced into myeloblastic cell line, T lymphocyte cell line and various tumor cell lines. Luciferase assay showed that APN upstream promoter is myeloid-specific for high expression in myeloblastic cell line and much lower expression in T lymphocyte cell line. The promoter activity was relatively high in lung adenoma cell line compared with other tumor cell lines including hepatoma cell line, tong cancer cell line and esophageal cancer cell line in which the promoter activity significantly diminished or was almost undetectable. The characteristics of APN promoter may provide a new strategy for specific myeloprotection while tumor patients are being treated with chemotherapy and/or radiotherapy.