Evaluation of stored red blood cell quality after washing using immune indices.

Washed red blood cells (RBCs) can be used to treat immune-related diseases. However, whether the washing process changes the quality of RBCs and affects the curative effect of transfusion therapy remains unclear. We retrospectively analysed the clinical data of patients who received blood transfusion. The physiological and biochemical parameters of RBCs were tested on an automated haematology-biochemical analyser. CD47 and phosphatidylserine (PS) plasma membrane expression were analysed using flow cytometry. Morphological changes in RBCs were observed using scanning electron microscopy. The results showed that the curative effect on patients who received washed RBCs was weaker than that on those who received non-washed RBCs. Physiological and biochemical parameters of RBCs were not significantly different. RBC immune indices changed significantly after washing. The expression of "don't eat me" signals was weakened, whereas the intensity of "eat me" signals was enhanced. This study suggests that the current use of physiological and biochemical parameters as indicators to evaluate the quality of RBCs may not be comprehensive and that evaluation of the real status of RBCs requires other effective parameters. Immune molecules in RBCs are expected to become supplementary markers for evaluating RBC quality.

Allogeneic platelet gel therapy for refractory abdominal wound healing: A preliminary study.

BACKGROUND: Refractory abdominal wounds are commonly complicated by surgical site infections, which prolong hospital stays and increase medical costs. There is little clinical data on the use of allogeneic platelet gel (PG) therapy for refractory infected wounds. OBJECTIVES: This study aimed to evaluate the efficacy and safety of allogeneic PGs in the treatment of refractory abdominal wounds. MATERIAL AND METHODS: A prospective single-center study was performed in a national abdominal trauma referral center between June 2019 and June 2021. A total of 11 patients with refractory abdominal wounds were treated with allogeneic PGs after the failure of standard medical treatments. The PGs were derived from platelets collected from healthy donors using apheresis, and each PG was tested for platelet count, transfusion-related diseases, aerobic and anaerobic bacteria, and growth factor concentration. Clinical efficacy was evaluated by assessing the wound surface and observing the condition of the wound, including wound area and percentage of granulation. RESULTS: The median age of the patients was 37 years (1st quartile, 3rd quartile (Q1, Q3): 31-55 years), median (Q1, Q3) hemoglobin level was 95 g/L (78-120 g/L) and median (Q1, Q3) serum albumin level was 39.9 g/L (34.9-42.7 g/L). The PG platelet count was 976.5 ±174.9×109/L. Results of transfusion-associated contagion tests for aerobic and anaerobic bacteria were negative. Growth factor contents (pg/mL) were: for transforming growth factor beta 1 (TGF-ß1); 2542.39 ±430.60, for platelet-derived growth factor BB (PDGF-BB); 23230.03 ±4236.14 and FOR vascular endothelial growth factor (VEGF); 91.41 ±23.31. The rate of wound healing was 100%, and the median (Q1, Q3) healing time was 30 days (18-40 days). The follow-up period was 5-27 months, during which no recurrence of the wounds was found. CONCLUSIONS: The present study demonstrated that allogeneic PGs are a safe and effective treatment for refractory abdominal wounds.

[Effects of Different Treatment Methods on the Contents of Related Growth Factors Released by Platelet Rich Plasma].

OBJECTIVE: To evaluate the effect of sonication, repeated freeze-thaw cycles, calcium salt solution and their combination on the content of related growth factors (GFs) released by platelet rich plasma (PRP). METHODS: Twenty PRPs from healthy blood donors were divided into 9 groups, including sonication group, freeze-thaw group, calcium gluconate group, calcium chloride group, sonication + calcium gluconate group, sonication + calcium chloride group, freeze-thaw + calcium gluconate group, freeze-thaw + calcium chloride group, and sonication + freeze-thaw group. After PRP activated by above 9 methods, the content of transforming growth factor-ß1 (TGF-ß1), vascular endothelial growth factor (VEGF), and platelet-derived growth factor-BB (PDGF-BB) were detected by ELISA. RESULTS: The platelet concentration of the samples was (966.7±202.6)×109/L. The content of TGF-ß1 in sonication + freeze-thaw group was the highest, while the lowest was in freeze-thaw group. The content of VEGF in freeze-thaw + calcium chloride group was the highest, while the lowest was in calcium gluconate group. The content of PDGF-BB in sonication + freeze-thaw group was the highest, while the lowest was in calcium gluconate group. There was no significant differences in the three GFs between calcium gluconate group and calcium chloride group. CONCLUSION: Among the 9 activated methods of PRP, there is no difference between two calcium salt solutions. And the combination of repeated freeze-thaw cycles and sonication may be the best treatment method to promote PRP to release GFs, while calcium gluconate is the weakest way.

GOx-Functionalized Platelet Membranes-Camouflaging Nanoreactors for Enhanced Multimodal Tumor Treatment.

Background: Glucose oxidase (GOx)-based starvation therapy is a new cancer treatment strategy. However, the characteristics such as limited curative effect and hypoxic tumor environment hinder its further application seriously. Methods: Herein, doxorubicin (DOX) loaded in hollow mesoporous copper sulfide (HMCuS) nanoparticles assembled with manganese dioxide (HMMD) as nanoshell was prepared. We developed a targeted enhanced cancer treatment method to camouflage HMMD by GOx-functionalized platelet (PLT) membranes (HMMD@PG). Results: GOx can be specially transported to the tumor site with PLT membrane for effective starvation treatment. Glucose and oxygen (O2) in the tumor were converted to H2O2 under the catalysis of GOx. HMMD can catalyze H2O2 to produce O2 and consume glutathione (GSH) in time, which regulates the tumor microenvironment (TME) and improves the adverse conditions of anti-tumor. In addition, DOX encapsulated in HMCuS-MnO2 release was accelerated from the nanoparticles after the "gatekeeper" MnO2 is consumed. The study of anti-tumor mechanism shows that the remarkable tumor suppressive ability of HMMD@PG comes from the three peaks synergy of starvation treatment, photothermal treatment (PTT), and chemotherapy. This nanoplatform disguised by PLT membrane has significant tumor inhibition ability, good biocompatibility and almost has no side effects in main organs. Conclusion: This work broadens the application mode of GOx and shows the new development of a multi-mode collaborative processing system of nanoplatforms based on cell membrane camouflage.

Seroprevalence of Transfusion Transmissible Infections and Associated Risk Factors in Hospitalized Patients before Transfusion in Jinling Hospital Nanjing University: A Three-Year Retrospective Study.

Transfusion-transmitted infections (TTIs), such as hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), and treponema pallidum (TP), must be detected before blood transfusion. However, few studies have been conducted on the prevalence and accuracy of positive results in hospitalized patients. The purpose of this study was to investigate the real seroprevalence of TTIs among patients before blood transfusion and analyze the characteristics of false-positive results in Jinling Hospital, Nanjing University, China. TTI results were collected from medical records and analyzed retrospectively. Additionally, we also used confirmatory assays to verify the accuracy of positive results. The overall prevalence of TTI was 8.96%, which was related to gender and age. The real positive rates were 86.67% (HBV), 35.09% (HCV), 20.75% (HIV), and 100% (TP). Our results also showed that high-speed centrifugation can reduce the false-positive rate of HBsAg. In summary, the results demonstrated that the positive rates of TTIs in hospitalized patients are higher than those in the general population. We also confirmed the existence of false-positive results in serological screening for TTIs. The method of processing specimens through high-speed centrifugation could reduce the false-positive results of detecting antigens effectively.

Current applications of platelet gels in wound healing-A review.

Human platelets play important roles in several physiologic and pathologic processes. Platelet concentrates are activated with thrombin or calcium, resulting in a viscous coagulum (platelet gel [PG]), composed of 95% platelets at least. PG is increasingly used for the treatment of a variety of soft and hard tissue defects, most notably in the management of chronic non-healing wounds. During wound healing, platelets not only play a critical role in primary hemostasis and thrombosis, but also release growth factors and cytokines to promote tissue regeneration, enhance collagen synthesis, and trigger an immune response. This review addresses a variety of aspects relevant to the functions of well-known platelet growth factors, animal and clinical studies of PG in the last decade, and different sources of platelets for PG. PG is used for non-healing chronic wounds, such as oral ulcerations related to epidermolysis bullosa and chronic graft-versus-host disease, for those, the traditional treatment effect is poor. PG maybe provide a new therapeutic direction for these diseases. Nevertheless, some uncertainty is present, the number of clinical studies is not enough. Hence, randomized controlled trials are still required to study the potential of the use of PG in the near future.

Serologic false-positive reactions for syphilis in children of adenoidal hypertrophy:2 case reports and review of the literature.

Adenoid hypertrophy (AH) is a common disease in children. We describe two cases of AH in children with syphilis enzyme-linked immunosorbent assay (EIA) for false-positive. To our knowledge, there are few reports of false-positive in syphilis EIA in children with AH. Two cases of AH children with syphilis EIA positive samples were confirmed by TPPA and LIA, both showed negative reaction. Therefore, the occurrence of syphilis EIA positive reactions in such diseases should dramatically arouse the attention of docimasters and doctors so as to avoid misdiagnosis caused by false-positive in children with AH.

Developments in the production of platelets from stem cells (Review).

Platelets are small pieces of cytoplasm that have become detached from the cytoplasm of mature megakaryocytes (MKs) in the bone marrow. Platelets modulate vascular system integrity and serve important role, particularly in hemostasis. With the rapid development of clinical medicine, the demand for platelet transfusion as a life­saving intervention increases continuously. Stem cell technology appears to be highly promising for transfusion medicine, and the generation of platelets from stem cells would be of great value in the clinical setting. Furthermore, several studies have been undertaken to investigate the potential of producing platelets from stem cells. Initial success has been achieved in terms of the yields and function of platelets generated from stem cells. However, the requirements of clinical practice remain unmet. The aim of the present review was to focus on several sources of stem cells and factors that induce MK differentiation. Updated information on current research into the genetic regulation of megakaryocytopoiesis and platelet generation was summarized. Additionally, advanced strategies of platelet generation were reviewed and the progress made in this field was discussed.

Progress in evaluating the status of hepatitis C infection based on the functional changes of hepatic stellate cells (Review).

Hepatitis C virus (HCV) infection is a global public health problem. Cirrhosis and hepatocellular carcinoma are the main causes of death in patients with chronic hepatitis C (CHC) infection. Liver fibrosis is an important cause of cirrhosis and end­stage liver disease after CHC infection. Along with the course of infection, liver fibrosis exhibits a progressive exacerbation. Hepatic stellate cells (HSCs) are involved in both physiological and pathological processes of the liver. During the chronic liver injury process, the activated HSCs transform into myofibroblasts, which are important cells in the development of liver fibrosis. At present, HCV infection still lacks specific markers for the accurate detection of the disease condition and progression. Therefore, the present review focused on HSCs, which are closely related to HCV­infected liver fibrosis, and analyzed the changes in the HSCs, including their surface­specific markers, cytokine production, activation, cell function and morphological structure. The present review aimed to propose novel diagnostic markers, at both the cellular and molecular level, which would be of great significance for the timely diagnosis of the disease. According to this aim, the characteristic changes of HSCs during HCV infection were reviewed in the present article.

An in vitro study of coagulation properties in refrigerated whole blood and reconstituted whole blood.

OBJECTIVE: Fresh whole blood (WB) has been used in military applications and cardiac surgery. We undertook a study of the coagulation properties of refrigerated WB stored for 21 days and compared them with the properties of reconstituted WB. STUDY DESIGN AND METHODS: Ten WB units were obtained from healthy volunteer donors and stored at 4 ± 2°C. Samples were obtained on Days 1, 2, 4, 6, 8, 10, 14 and 21 from the WB units. Ten units of reconstituted WB were prepared with a ratio of red cells, platelets and plasma of 1:1:1. Tests included complete blood count, electrolyte, routine coagulation, blood coagulation factor and thromboelastography. RESULTS: There was a progressive decline in Hb, WBC, PLT, sodium and coagulation factors but a progressive increase in APTT, PT and potassium in WB. The concentrations of factor (F)V and FVIII as well as FII and FX of WB were higher before Days 4, 2, 8 and 14, respectively, compared with the concentrations of reconstituted WB. The concentrations of FVII, FIX, FXI and FXII in WB were found to be equal to or higher than those in reconstituted WB throughout the course of 21 days. TEG variables in all WB units were normal throughout the course of 10 days. The mean PT and APTT of WB were lower than those of reconstituted WB before Days 14 and 10, respectively. CONCLUSION: This study suggests that the coagulation properties of refrigerated WB were equal to or superior to those of reconstituted WB for a minimum of 10 days.

[Coagulation Properties for Whole Blood Stored at 4℃].

OBJECTIVE: To study the coagulation properties the refrigerated whole blood stored at 4℃. METHODS: Ten units of whole blood were obtained from healthy volunteer donors and stored at 4±2℃ for 21 days. Samples were collected on the day after donation and on days 2, 4, 6, 8, 10, 14 and 21 for delection including complete blood count, electrolyte, APTT, PT, Fg, blood coagulation factors, and thromboelastography(TEG). RESULTS: The levels of Hb, WBC, Plt, sodium and potassium in each sample accorded with standard of storing whole blood. The level of Hb, WBC, Plt and Na+ decreased along with prolonging of storage time, while the K+ level increased along with prolonging of stored time, APTT and PT prolonged along with prolonging of thored time, PT>17 min at d 21, the Fg level change was no-obvious, The level of factor Ⅴ and Ⅷ decreased more than 50 % of baseline on d 6 and 4 respectively; the levels of factor Ⅱ, Ⅶ, Ⅸ, Ⅹ, Ⅺ, Ⅻ showed decreasing trend, but their levels were less than 40 % of baseline values at d 21. TEG test showed that no abnormalily of R value was found, the abnormal valnes of K and Angle were observed at d 21, the abnormal value of MA was observed at d 14. CONCLUSION: The whole blood stored for 10 days possesses normal coagulation function showing important significance for treatment of hemorrhage from war injury and surgical openation of heart and chest.

Advancement of platelet-inspired nanomedicine.

Human platelets play important roles in several physiologic and pathologic processes. Any defects in platelet number and function can lead to a variety of bleeding complications; platelet transfusions are commonly used to improve platelet count and hemostatic function in these cases. However, transfusions are complicated by the short storage life of donated platelets and contamination risks. Synthetic platelet substitutes can render efficient hemostasis while allowing advantages of large-scale preparation, minimum contamination risk via effective sterilization, longer shelf-life, no need for blood type matching, and reduced risks of biologic or pathologic side effects. Recent efforts in synthetic platelet design have additionally focused on applications including hemostasis, drug delivery, and targeted delivery of cancer therapeutics. In the present review, we summarize the advancement of platelet-inspired nanomedicine for artificial blood component, tumor research, and treatment of thrombi.

Platelet-inspired medicine for tumor therapy.

As the number of patients with tumor increases dramatically recent years, traditional therapies expose more and more problems which can even lead to death. Many researchers and clinicians quest for an efficient drug delivery system to deal with tumor as a result. With the researches further develop, we find that platelet can interact with tumor cells through a variety of ways. So it can be used as a carrier broadly to deliver different anti-tumor drugs for tumor treatment. In the present review, we summarize the interaction of tumor cells and platelet. At the same time, we focus on recent progress on the application of platelet drug-loaded system in the anti-tumor prospects.

A case of dispermic chimerism with normal phenotype identified during ABO blood grouping.

BACKGROUND: Human chimerism with normal phenotype derived from the fusion of two different zygotes is a rare phenomenon. We describe a case of a phenotypically normal 17-year-old diagnosed with dispermic chimerism during routine ABO blood grouping. METHODS: ABO grouping, ABO genotyping, karyotyping, human leukocyte antigen (HLA) typing, and short tandem repeat (STR) analysis were performed. RESULTS: Forward typing with anti-A and anti-B sera resulted in mixed-field agglutination of red blood cells. The mother and father were blood group O and AB, respectively. The proposita had O1, A201 and B alleles in the ABO locus; O1 was a maternal allele, while A201 and B were the paternal alleles. The proposita karyotype was 46,XX/46,XY. HLA typing revealed that the proposita had three alleles (46, 51, 54) at the HLA-B locus, with the additional allele of paternal origin. STR analysis identified three alleles for five of the 15 markers (D2S1338, TPOX, D8S1179, D19S433, and D21S11) analyzed in the proposita's blood- and skin fibroblast-derived DNA. The additional alleles of TPOX, D8S1179, and D21S11 were of paternal origin. CONCLUSIONS: The genetic findings suggest that this proposita was produced by dispermic fertilization of two identical haploid ova formed by parthenogenetic activation.

Biocompatibility, biodegradation, and neovascularization of human single-unit platelet-rich fibrin glue: an in vivo analysis.

BACKGROUND: The clinical applications of fibrin glue span over several surgical modalities. The aim of this study was to evaluate the biocompatibility and biodegradation of different formulations of platelet-rich fibrin glue in vivo and examine its effects on the neovascularization of wound sites. METHODS: Human-derived single-unit fibrin glue was prepared. Incisions were made on the backs of rats, and these were coated with homemade glues containing different concentrations of aminomethylbenzoic acid (Groups A-F) or commercial adhesives (Group G). A sham control group was included (Group H). The wounds were examined by histological analysis and immunohistochemistry at several time points. RESULTS: Successful wound closure was achieved in all groups by day 12. Acute inflammation occurred during the first six days, but gradually disappeared. The longest sealant duration was achieved using the lowest concentration of anti-fibrinolytic agent in a 1:10 volume ratio with cryoprecipitate. Expression levels of the platelet endothelial cell adhesion molecule-1 were significantly higher in Groups A and C compared to the control groups (Groups G and H) on day 3 (P < 0.05). CONCLUSIONS: Single-unit platelet-rich fibrin glue has excellent biocompatibility and is associated with the upregulation of neovascularization. The addition of aminomethylbenzoic acid could prevent the degradation of fibrin glue.

Antimicrobial properties of single-donor-derived, platelet-leukocyte fibrin for fistula occlusion: An in vitro study.

Fibrin glue is a promising alternative for low-output enterocutaneous fistula closure. Bacterial flora colonizing inside the fistula tract, however, may limit the glue application. Single-donor-derived, platelet-rich materials were hypothesized in this study to have antimicrobial activity against Gram-negative microorganisms. Platelet-leukocyte fibrin (PLF), platelet-rich plasma (PRP), and platelet-poor plasma (PPP) were obtained from healthy volunteers. The amounts of platelet, leukocyte, and complement/antibody were determined. In vitro laboratory susceptibility to PLF and plasmas was determined by the Kirby-Bauer disc-diffusion method. Antimicrobial activity of PLF, PRP, and PPP against three Gram-negative ATCC strains was determined in a bacterial kill assay. Levels of complement and antibody did not significantly differ among PLF, PRP, and PPP (p > 0.05), while platelet and leukocyte counts in platelet-rich biomaterials were significantly higher than those in PPP (p < 0.001). The bactericidal effects produced by PLF and PRP were more pronounced than those by PPP in both qualitative and quantitative aspects. No inhibitory zones against three Gram-negative organisms were observed in the Bioseal®. In conclusion, our homemade PLF, as compared with commercial products, had remarkable antimicrobial activity against Gram-negative bacteria relevant to fistula colonization. The presence of platelets and leukocytes may play an important role in bacterial defense. This is the first study to demonstrate the antibacterial properties of single-unit PLF for fistula closure, presenting a new opportunity for glue sealing.

Preparation of PNIPAM-g-P (NIPAM-co-St) microspheres and their blood compatibility.

The poly(N-isopropylacrylamide)-g-poly(N-isopropylacrylamide-co-styrene) microspheres (PNNS-MSs) were prepared by an emulsifier-free emulsion polymerization method. The blood compatibility of PNNS-MSs was characterized by in vitro for coagulation tests, hemolysis assay, plasma recalcification time, complement activation, platelet activation, and cytotoxicity experiments. The results showed that the PNNS-MSs have good blood compatibility and lack cytotoxicity, which may be attributed to the formation of a strong interfacial hydration layer that result from amphiphilic molecular structure of the PNIPAM shell and minimal interaction between PNNS-MSs interfaces and blood components. The PNNS-MSs provide a promising platform of blood circulation system for early illness diagnosis and therapy.

Hemocompatible and antibiofouling PU-F127 nanospheres platform for application to glucose detection in whole blood.

Analysts are always enthusiastic about finding new materials with good biocompatibility to improve the behavior of biosensors. In this case, a novel electrochemical biosensor, which can be conveniently applied to veraciously evaluate the level of blood glucose with the help of antibiofouling technology, was prepared and investigated. More details of the preparation of polyurethane-Pluronic F127 (PU-F127) nanospheres and immobilizing glucose oxidase (GOx) on (PU-F127)-glass carbon electrode (GCE) were presented. The electrochemical behavior of the biosensor in whole blood was studied. The cyclic voltammetric results indicated that GOx immobilized on the PU-F127 nanospheres exhibited direct electron transfer reaction, which led to stable amperometric biosensing for glucose with a detection limit of 1.14 × 10-5 M (S/N = 3) in whole blood. The PU-F127 nanospheres modified GCE also offered good anti-interference ability to ascorbic acid (AA) and uric acid (UA), especially when a detection potential of -0.49 V was employed. The good stability and repeatability of this biosensor were also proved. The integration of the technologies, which include anticoagulants, sensors and nanoscience, will have significant input to high-performance biosensors relevant to diagnostics and therapies of interest for human health.

Preparation, blood compatibility and anticoagulant effect of heparin-loaded polyurethane microspheres.

Recent advances in micro- and nanotechnology have provided a variety of particles with highly controlled shapes, sizes, chemical composition, and interesting properties. In this work, a novel kind of heparin-functionalized polyurethane microsphere (Hep-PU MS) was synthesized by a single-step phase separation method. The blood compatibility and anticoagulant effect of the Hep-PU MSs were investigated using coagulation tests, hemolysis assay, complement and platelet activation detection, cytotoxicity analysis, and drug loading and release study. The results confirmed that the heparin can substantially enhance the anticoagulant properties of PU MSs, and the Hep-PU MSs have the potential to be used as a mild anticoagulant compared to heparin. With the simplicity of the functionalized method, the excellent hemocompatibility and the slow-release of heparin, the Hep-PU MSs with desirable bioproperties can be readily tailored to cater to various biomedical applications.

Biochemical, mechanical, and morphological properties of a completely autologous platelet-rich wound sealant.

The optimal composition of fibrin sealant preparations is not known. We, therefore, sought to construct a series of sealants from autologous components and compare their functioning to Bioseal a commonly used sealant. Characteristics of the platelet-rich plasma, cryoprecipitate, and thrombin were determined and compared to commercial glue from composition, function, and microscopy aspects. The concentrations of platelets as well as fibronectin in autologous fibrin glues were significantly higher than those in commercial ones (P < 0.001). Mechanical values (maximum amplitude and clot strength) obtained from thrombelastograph assays in two groups were not significantly different (P > 0.05). A dense platelet surface and fibrin net structures could be observed in the autologous samples, whereas there were only sparse fibrin nets without cellular components in Bioseal. Characterization of autologous and Bioseal fibrin sealants and their performance do not have significant difference in biochemical and mechanical properties. The entirely autologous platelet-rich gel in the present study may get wide application in future practice after confirmation of its safety, efficiency, and economic benefits.