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Thromb Haemost ; 120(4): 647-657, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32289862


Heparanase (HPSE) is an endo-ß-D-glucuronidase that cleaves heparan sulfate and hence participates in remodeling of the extracellular matrix, leading to release of cytokines that are immobilized by binding to heparan sulfate proteoglycans (HSPGs), and consequently activating signaling pathways. This function of HPSE is correlated to its expression level that is normally very low in majority of the tissues. Exceptionally, human platelets express high level of HPSE, suggesting a unique physiological role in this cell. Using K562 cell line, we found a progressive increase of HPSE during the megakaryocytic differentiation. Analysis of a series of megakaryocytic differentiation-related heparin-binding proteins (HBPs) in the cell culture medium revealed an exclusive positive correlation between the level of interleukin 6 (IL-6) and HPSE expression. IL-6 modulated megakaryocytic differentiation through activation of STAT3. Further, we demonstrated that overexpression of HPSE potentiates megakaryocytic differentiation, whereas elimination of HPSE led to a delayed differentiation. This function of HPSE is associated with its activity, as overexpression of inactive HPSE had no effect on IL-6 production and megakaryocytic differentiation. The role of HPSE is further supported by the observation in an umbilical cord blood CD34+ cells megakaryocytic differentiation model. Our data propose a novel role for HPSE in platelets production by a HPSE/IL-6/STAT3 positive feedback loop that specifically regulates megakaryocytes maturation.

Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(1): 296-299, 2020 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-32027292


OBJECTIVE: To establise the bank of platelet donors with the human platelet antigen (HPA) 1-6, 15 genes so as to provide the HPA-matched platelets for the patients. METHODS: The HPA genotyping of platelets donors and patients with platelet antibody positive confirmed by sercening was performed by using the SSP-PCR; the efficacy of transfusing the HPA-matched platelets for 37 cases platelet antibody positive was analyzed. RESULTS: The most common genotype in platelet donors were HPA-1a/1a-2a/2a-3a/3b-4a/4a-5a/5a-6a/6a-15a/15b, followed by HPA-1a/1a-2a/2a-3a/3a-4a/4a-5a/5a-6a/6a-15a/15b; the most common genotype in 53 cases of platelet antibody positive confirened by screening were HPA-1a/1a-2a/2a-3a/3b-4a/4a-5a/5a-6a/6a-15a/15b. Among 37 patients with platelet antibody positive confirened by screeming, 28 showed that the transfusion of HPA-matched platelets was effective with statistically significant difference in comparison with random transfusion group. The HPA-3, HPA-15 were the main factors leading to polymorphisms. CONCLUSION: HPA-3 and HPA-15 are polymorphic, which should be focused on. HPA-matched platelets can improve the efficiency of platelet transfusion, and avoid the waste of blood resources. The genotypes of platelet donors can basically meet the requirements for common genotype transfusion.

Plaquetas , Antígenos de Plaquetas Humanas , Doadores de Sangue , Frequência do Gene , Genótipo , Humanos , Polimorfismo Genético
Blood Transfus ; 14(2): 168-74, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26509826


BACKGROUND: Enzymatic conversion of blood group A1B red blood cells (RBC) to group O RBC (ECO) was achieved by combined treatment with α-galactosidase and α-N-acetylgalactosaminidase. The aim of this study was to evaluate the function and safety of these A1B-ECO RBC in vitro. MATERIALS AND METHODS: A 20% packed volume of A1B RBC was treated with enzymes in 250 mM glycine buffer, pH 6.8. The efficiency of the conversion of A and B antigen was evaluated by traditional typing in test tubes, gel column agglutination technology and fluorescence-activated cell sorting (FACS) analysis. The physiological and metabolic parameters of native and ECO RBC were compared, including osmotic fragility, erythrocyte deformation index, levels of 2,3-diphosphoglycerate, ATP, methaemoglobin, free Na(+), and free K(+). The morphology of native and ECO RBC was observed by scanning electron microscopy. Residual α-galactosidase or α-N-acetylgalactosaminidase in A1B-ECO RBC was detected by double-antibody sandwich ELISA method. Manual cross-matching was applied to ensure blood compatibility. RESULTS: The RBC agglutination tests and FACS results showed that A1B RBC were efficiently converted to O RBC. Functional analysis suggested that the conversion process had little impact on the physiological and metabolic parameters of the RBC. The residual amounts of either α-galactosidase or α-N-acetylgalactosaminidase in the A1B-ECO RBC were less than 10 ng/mL of packed RBC. About 18% of group B and 55% of group O sera reacted with the A1B-ECO RBC in a sensitive gel column cross-matching test. DISCUSSION: The conversion process does not appear to affect the morphological, physiological or metabolic parameters of A1B-ECO RBC. However, the A1B-ECO RBC still reacted with some antigens. More research on group O and B sera, which may partly reflect the complexity of group A1 the safety of A1B-ECO RBC is necessary before the application of these RBC in clinical transfusion.

Sistema ABO de Grupos Sanguíneos/química , Tipagem e Reações Cruzadas Sanguíneas/métodos , Eritrócitos/química , Humanos , Masculino
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 23(4): 1152-5, 2015 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-26314463


OBJECTIVE: To evaluate the storage performance of the domestically made platelet storage bags (experimental group) and the United States Trima set platelet storage bags (control group). METHODS: The manually separated platelets were divided in two equal parts, which was added to control blood bags and experimental blood bags respectively, all samples were stored at a 22 °C ± 2 °C. The platelet count, mean volume, aggregation activity (ADP, THR), pH, glucose, lactate concentration, lactate dehydrogenase concentration, hypotonic shock reaction, CD62P and phosphatidic acid serine content were detected at day 0, 3, 5 and 7 of storage. RESULTS: There was no significant difference of platelet quality at day 5 after storage between the experimental group and the control group (T-test, P > 0.05). CONCLUSION: Two kinds of platelet storage bags have the similar storage performance.

Plaquetas , Preservação de Sangue , Separação Celular , Glucose , Humanos , Contagem de Plaquetas
Cell Rep ; 7(6): 1982-93, 2014 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-24882011


CUEDC2, a CUE-domain-containing protein, modulates inflammation, but its involvement in tumorigenesis is still poorly understood. Here, we report that CUEDC2 is a key regulator of macrophage function and critical for protection against colitis-associated tumorigenesis. CUEDC2 expression is dramatically upregulated during macrophage differentiation, and CUEDC2 deficiency results in excessive production of proinflammatory cytokines. The level of CUEDC2 in macrophages is modulated by miR- 324-5p. We find that Cuedc2 KO mice are more susceptible to dextran-sodium-sulfate-induced colitis, and macrophage transplantation results suggest that the increased susceptibility results from the dysfunction of macrophages lacking CUEDC2. Furthermore, we find that Cuedc2 KO mice are more prone to colitis-associated cancer. Importantly, CUEDC2 expression is almost undetectable in macrophages in human colon cancer, and this decreased CUEDC2 expression is associated with high levels of interleukin-4 and miR-324-5p. Thus, CUEDC2 plays a crucial role in modulating macrophage function and is associated with both colitis and colon tumorigenesis.

Proteínas de Transporte/metabolismo , Neoplasias do Colo/metabolismo , Macrófagos/metabolismo , Proteínas de Membrana/metabolismo , MicroRNAs/metabolismo , Proteínas Repressoras/metabolismo , Animais , Proteínas de Transporte/biossíntese , Proteínas de Transporte/genética , Proteínas de Transporte/imunologia , Colite/genética , Colite/imunologia , Colite/metabolismo , Colite/patologia , Neoplasias do Colo/genética , Neoplasias do Colo/imunologia , Feminino , Regulação da Expressão Gênica , Células HeLa , Humanos , Ativação de Macrófagos , Macrófagos/imunologia , Macrófagos/patologia , Proteínas de Membrana/biossíntese , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Camundongos , Camundongos Transgênicos , MicroRNAs/genética , Proteínas Repressoras/biossíntese , Proteínas Repressoras/genética , Proteínas Repressoras/imunologia , Transdução de Sinais
Transfusion ; 53(4): 868-71, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22882577


BACKGROUND: Compared to ISBT128 code labels, radiofrequency identification (RFID) tags have incomparable advantages and gradually applied in blood management system. However, there is no global standard for the uses of RFID frequency. Even though ISBT recommended high-frequency RFID with 13.56MHz, 820- to 960-MHz ultrahigh frequency (UHF) RFID technology in many ways has even more advantages. For this reason, we studied the effect of UHF RFID tags with 820- to 960-MHz exposure on storage quality of red blood cells (RBCs) and platelets (PLTs). STUDY DESIGN AND METHODS: Thirty units of collected and prepared suspended RBCs (sRBCs) and PLTs were divided into two bags, one each for the test and control groups. The sRBCs were stored in 4±2°C refrigerator and the PLTs in a 22±2°C rocking box. The test groups were exposed to RF reader continuously during storage. Sampling at different time points and biologic changes were tested. RESULTS: As the extension of storage and the pH and chlorine levels in the supernatant of sRBCs were reduced, free hemoglobin, potassium, and sodium increased, but were not significant between test and control groups (p>0.05). During the storage period, the pH levels, PLT count, and PLT aggregation rate were decreased in both test and control groups, but were not significant (p>0.05). CONCLUSION: When exposed to 820- to 960-MHz RF, the biologic and biochemical indexes are not found to be exacerbated during 35 days of storage for sRBCs and 5 days for PLTs, respectively.

Plaquetas/efeitos da radiação , Preservação de Sangue/métodos , Eritrócitos/efeitos da radiação , Rotulagem de Produtos/métodos , Ondas de Rádio , Plaquetas/fisiologia , Preservação de Sangue/instrumentação , Índices de Eritrócitos , Eritrócitos/fisiologia , Humanos , Agregação Plaquetária , Contagem de Plaquetas , Rotulagem de Produtos/instrumentação