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【Objective】 To analyze the data of clinical blood transfusion quality control supervision in Shanghai, so as to provide reference for the improvement of clinical blood transfusion quality management in hospitals at all levels. 【Methods】 The data of clinical blood transfusion quality control supervision in hospitals at all levels from 2016 to 2021 were retrospectively analyzed to obtain the characteristics and indicators in the quality management. 【Results】 The overall level of clinical blood transfusion quality management in Shanghai steadily improved from 2016 to 2021 (F=3.82, P<0.01), and the management level of different hospitals varied significantly (F=9.00, P<0.01). In 2021, the full compliance rates of housing facilities, instruments and equipment, diagnostic reports and medical record writing among the third-level indicators of clinical blood transfusion quality management in hospitals at all levels were as follows: 86.49%(32/37), 100% (37/37)and 43.24%(16/37) for tertiary comprehensive hospitals; 61.11%(11/18), 88.89%(16/18) and 50.00% (9/18)for tertiary specialized hospitals; 60.87%(14/23), 78.26%(18/23)and 47.83%(11/23) for secondary comprehensive hospitals, ; 60.00%(9/15), 66.67%(10/15), 40.00%(6/15) for secondary specialized hospitals; 52.38%(11/21), 38.10%(8/21), 42.86%(9/21) for private hospitals. 【Conclusion】 The characteristics of clinical blood transfusion quality management in hospitals at all levels in Shanghai differed significantly, with different strengths and weaknesses. Hospitals should improve blood transfusion management in terms of housing facilities, personnel management, system process as well as diagnostic reports and medical record writing, in order to enhance the clinical blood transfusion quality management.
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In order to solve the difficulties and challenges in the implementation of the original blood distribution and collection regulations caused by the expansion of hospital area, the extension of blood transfer time, the changeability of blood transfer environment, and the strain of personnel due to the increase of workload, as well as to ensure the accuracy of the information throughout blood remote verification and distribution and the safety of clinical blood transfusion, , Shanghai experts related to clinical transfusion and blood management had made a systematic study on the applicable scope and management rules of remote verification of blood distribution and collection, and formulated this Expert Consensus combined with the development status of digital, intelligent and remote communication technologies, so as to provide corresponding guidance for clinical medical institutions in line with the changes in reality.
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【Objective】 To optimize the existing spin-EB method and promote human induced pluripotent stem cells (hiPSCs) differentiate into megakaryocytes (MKs). 【Methods】 In this study, the initial inoculation amount of hiPSCs was increased from 3 500 cells/well to 8 000 cells/well, and the size of EB was increased. By observing the generation time of EB- hematopoietic cells during differentiation, and detecting the proliferation of CD34+ hematopoietic progenitor cells and CD41+ MKs in different stages, it was studied whether the optimized scheme could promote the differentiation of hiPSCs into hematopoietic progenitor cells(HPCs) and MKs. 【Results】 By increasing the initial inoculation amount of hiPSCs and the size of EB, the differentiation of hiPSCs into HPCs and MKs and the cell production efficiency can be promoted. 【Conclusion】 Our research describes an optimized and repeatable differentiation method, which can produce hematopoietic progenitor cells and mature MKs from hiPSCs in a relatively short time with higher yield. It is of great clinical significance and broad scientific research prospect to continuously optimize the culture scheme of hiPSCs differentiation to produce MKs and platelets in vitro, and to promote large-scale platelet generation in vitro in transfusion medicine.
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With continuous discovery of tumor immune targets and continuous changes in antibody research and development technology, antibody drugs are becoming more and more widely used in clinical practice. However, some targets are not only expressed on tumor cells, but also on red blood cells. Therefore, the clinical application of antibodies against the corresponding targets may interfere with the detection of blood transfusion compatibility, resulting in difficulty in blood matching or delay of blood transfusion. This consensus summarizes the current solutions for the interference of CD38 monoclonal antibody (CD38 mAb) in transfusion compatibility testing. After analyzing the advantages and disadvantages of different methods, polybrene and sulfhydryl reducing agents [dithiothreitol (DTT) or 2-mercaptoethanol (2-Me)], as a solution for CD38 mAb interference in blood compatibility testing, are recommended for Chinese patients, so as to eliminate blood transfusion interference produce by CD38 mAb and further provide a pre-transfusion workflow for clinicians and technicians in Department of Blood Transfusion.
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Platelet transfusion is the main clinical treatment of thrombocytopenia. However, due to the difficulty of platelet collection, high cost of use and limited number of blood donors, the development of platelet treatment is greatly limited. Therefore, the research on thrombogenesis in vitro has attracted more attention at home and abroad. Platelet production in vitro has the advantages of donor-independence, platelet antigen free and low risk of alloimmunity. At present, the efficiency of producing functional platelets in vitro is low, and there is still a big gap to achieve the ultimate goal of producing a large number of functional platelets in vitro. This paper reviews the research progress of megakaryocyte / platelet production in vitro, focuses on the in vitro production potential of megakaryocyte / platelet, and summarizes the current platelet culture systems in vitro based on human pluripotent stem cells, embryonic stem cells and adipose stem cells. The contradictions and difficulties of platelet production in vitro were also discussed to provide theoretical support for further research.
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【Objective】 To investigate the effect of cold agglutination on blood group typing. 【Methods】 37℃ water bath, absorption elution test and 2-mercaptoethanol method were used to eliminate the influence of cold agglutination. Forward and reverse blood group typing, cross matching, DAT and IAT experiments were then performed on red blood cells and serum after treatment. 【Results】 Before treatment, obvious discrepancy in forward /reverse typing and nontypable cross matching in 16 blood samples were noticed due to cold agglutination. After corresponding treatments, all samples were consistent or negative in forward/reverse typing, cross matching and antibody screening. No adverse reactions to cross matching blood transfusion occurred in patients, and the increase of hemoglobin was in line with the effective standard of transfusion. 【Conclusion】 37℃ water bath, absorption elution test and 2-mercaptoethanol method can be used to eliminate the interference caused by cold agglutination to obtain correct typing results. The strong reactivity caused by cold agglutination in AIHA patients were different from other cases, which deserved our attention.
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This study aimed at developing restraint standards for quality and safety of blood transfusion services for blood transfusion administration in clinical blood use, as well as such issues before, during and after transfusion. Such means as literature analysis, blood transfusion adverse event analysis, and system standard interpretation were used to study these issues found in clinical blood use at home and abroad, with key points identified according to the universality, high-prevalence rate, criticality and impact degree. A standard framework is established centering on transfusion service and key points as nodes. Hence a standard text comes into being, comprising four sections and 20 key points.
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Objective To analyse the effect of specimen storage time on platelet count in apheresis Donors,in order to choose the appropriate testing time.Methods We choose fifty healthy Unpaid Blood Donors of Platelets to test platelet count in 0,0.5,1,3,6h respectively by the blood counting instrument.Results At room temperature,the count of platelets from blood samples is relatively lower at 0 hour and the difference is significant (P ＜ 0.05).After 0.5-6 hours,the count of platelets become stabilized and not has significantly different(P＞0.05).Conclusion The count of platelets of blood samples is lower at 0 hour than 0.5-6 hours,this work suggest that the count of platelets of blood samples should be done at 0.5-6 h in order to protect platelets quality.
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Objective To investigate the status of blood donation in Shanghai streets,understanding the motivation of the donors for blood donation and how many times they had donate their blood.Here,we intend to improve work efficiency in order to better serve the blood donators and to provide a theoretical basis.Methods To gain insight into the understanding and attitude of citizens for blood donation,we analyzed the volunteer blood donors in 387 vehicles and 436 outdoor pre mobilization object by six months,questionnaire in blood collection vehicle or at scene etc.Results The survey specifically reflected the understanding,attitude and motivation of citizens and foreign workers for blood donation in Shanghai.Conclusion There existed many problems such as wrong recognition,diversified motivation among many citizens for blood donation.More publicity and education for the knowledge of blood donation are needed and it plays significant and longlasting role for blood donation.
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<p><b>OBJECTIVE</b>To improve the MigR1-CD19-CAR (chimeric antigen receptor) that contains a single chain variable region (scFv) which targeted to CD19 through a retroviral vector transduction efficiency of T-lymphocytes.</p><p><b>METHODS</b>Insert the CD19-CAR fragment into the retroviral vector (MigR1) through recombinant DNA technology, after transfecting plat-A packaging cell lines, viral supernatant was collected to transduce K562 cell line and activated human T-lymphocytes. We used flow cytometry to determine the transduction efficiency and RT-PCR to confirm the transcription of CD19-CAR gene. The ability of the transduced T cells to produce IFN-γ and TNF-α in a CD19-specific manner was measured in an enzyme-linked immunosorbent (ELISA) assay.</p><p><b>RESULTS</b>(1)Using MigR1-CD19-CAR retroviral vector to produce the high titer retrovirus. (2)MigR1-CD19-CAR transduction efficiency of K562 cell line was significantly higher than human T-lymphocytes (P<0.01). (3)120 min centrifugation could significantly improve transduction efficiency of T-lymphocytes to (54.5±14.6)%. (4)Transduction efficiency could be improved by deciding transduce time according to T-lymphocytes proliferation fold in vitro individually, and the highest transduction efficiency in the study was 69.3%. The CD19-CAR gene sequence was transcripted specificly with high efficiency. (5) IFN-γ and TNF-α released by CD19-CAR transduced T-lymphocytes significantly increased to (13 230±1 543) pg/ml and (4 217±211) pg/ml when coculture with CD19-K562 cells.</p><p><b>CONCLUSION</b>We have successfully constructed a second generation CAR which targeted to CD19 through a retroviral vector called MigR1 (MigR1-CD19-CAR). Deciding transduce time according to T-lymphocytes proliferation fold in vitro individually and 120 min centrifugation could improve the CAR transduction efficiency of T-lymphocytes. RT-PCR confirmed that the CD19-CAR gene was specificly transcripted with high efficiency. IFN-γ and TNF-α released by CD19-CAR transduced T-lymphocytes significantly increased when activated by target cells.</p>
Subject(s)
Humans , Antigens, CD19 , Cell Proliferation , Flow Cytometry , Genetic Vectors , K562 Cells , Recoverin , Retroviridae , T-Lymphocytes , TransfectionABSTRACT
Objective To explore TRAIL expression in patients with primary immune thrombocytopenia (ITP)and its role in the pathogenesis.Methods Peripheral Blood Mononuclear Cells (PBMCs)from twenty-eight patients with ITP and thirty healthy controls were obtained.TRAIL expression was determined using Real-time Quantitative Polymerase Chain Reaction (RT-PCR).Serum TNF-α,IFN-γ,IL-4 and IL-10 were detected using ELISA.Associations between TRAIL expression and clinical parameters were assessed.Results Compared to healthy controls,TRAIL expression was significantly increased in PBMC from patients with ITP (2.80±0.43 vs 1.00±0.24,t=19.72,P0.05).Furthermore,TRAIL expression was positively correlated with serum IFN-γ,TNF-αlevel (r=0.432,0.541,P<0.05),and negatively correlated with IL-10 and PLT (r=-0.424,-0.553,P<0.05).Conclusion Rur results suggest that TRAIL may be involved in the pathogenesis of ITP.
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Objective To establish a off-pump leukocyte depletion method in canines.Methods Twenty-one healthy adult mongrel dogs of both sexes, weighing 10-12 kg, were randomized into 2 groups: control group (group C, n = 9) and leukocyte depletion group (group LD, n = 12) . An extracorporeal leukocyte filtration end was constructed by aseptic puncture of the bilateral external jugular veins via a blood transfusion line and the leukocyte filter for extracorporeal leukocyte depletion. In group LD, blood was filtered with a MP-300 blood line and a SQ40S leucocyte depletion filter one end placed in the right external jugular vein (artery end) and the other end placed in the left external jugular vein (venous end). After heparin anticoagulation, a MP-300 blood line pump was used as the power. Blood was filtered at a rate of 75 ml/min and it was maintained for 60 min. The artery end was then closed, normal saline injected into the closed circuit, the remaining blood pumped into the body, and then the venous end closed. In group C, aseptic puncture of the bilateral external jugular veins was performed. Arterial blood samples were taken immediately before leukodepletion (baseline, T_0 ) , at 10, 20, 30, 40, 50 and 60 min of leukodepletion (T_(1-6) , depletion period), and at 10, 20, 30, 40, 50, 60, 120, 180 and 270 min after leukodepletion (T_(7-15) , recovery period) for determination of blood routine. MAP, HR, RBC.Plt and body temperature were recorded at T_0 , T_6 , T_(12) and T_(15) . Results There were no significant difference in MAP and RBC between the two groups ( P > 0.05 ) . HR, body temperature, and Plt were significantly lower in group LD than in group C (P < 0.05) .The leukocyte concentration was lower during depletion period in group LD than in group C ( P < 0.05) , while there was no significant difference in leukocyte concentration during recovery period between the two groups (P > 0.05 ). Conclusion The off-pump leukocyte depletion method is successfully established and has exact efficacy with less adverse effects in this study.
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Objective To study complement receptor typeⅠ(CR1)activity of erythrocytes in pa-tients with SLE.Methods Using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP),CR1rosette(RBC-CR1R)and immuno-complex rosette(RBC-ICR)on red cell,the ery-throcyte complement receptor I type(ECR1)genomic density polymorphism(HH type,HL type,LL type)and erythrocyte CR1immune activity were determined in32patients with SLE and in48normal individuals.Results It was found that HH type rate of ECR1density polymorphism in patients with active SLE was significantly lower(10/16,62.5%)than that(13/16,81.3%)in patients with stable SLE.The level of CR1immune activity in HH type was significantly higher than that in HL,LL type of SLE,and significantly dif-ferent from that in48normal individuals(P
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Objective To explore the method and feasibility of preparing MTX+VCR-loaded erythrocytes using healthy adult erythrocytes. Methods We used the modified hypotonic pre-swelling and isotonic resealing technique to prepare MTX+VCR-loaded erythrocytes. The loading parameters and releasing rate were evaluated with high penformance liquid chromatograph (HPLC) and auto-cytoanalysis. We observed the osmotic fragility of MTX+VCR-loaded erythrocytes conventionally. The change in innate immune activity of co-encapsulated erythrocytes was measured by counting the rosette rate of MTX+VCR-loaded erythrocytes adhering to S180. Results The concentration of MTX showed significant influence on the co-encapsulation efficiency (P0.05). Conclusions The preparation and biological characteristics of MTX+VCR-loaded erythrocytes were not different from those of mono-loaded erythrocytes, and erythrocytes were practically capable to be used as carriers for co-encapsulation of MTX and VCR.
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Objective To study the effect of Vincristine-loaded intact human erythrocytes on tumor cell in vitro. Methods VCR was loaded in intact human erythrocytes using a modified hypotonic preswelling technique. The proliferation of K562 cells after incubation with VCR-loaded erythrocytes was detected by MTT. Cellular cycle and apoptosis of K562 cells were analyzed through FACS staining with PI/Rhodamin123, and morphology of apoptotic K562 cells and their nucleus were observed through staining with Hoechest33258. Results VCR-loaded erythrocytes could efficiently inhibit the proliferation of co-cultured K562 cells and induce the cells′ apoptosis in a dosage-dependent manner. Cellular cycle analysis demonstrated that proportion of K562 cells in G 0/G 1 decreased whereas in S+G 2/M increased after incubation with VCR-loaded erythrocytes. Such effects could also be displayed from alone VCR group, and significant change was observed in K562 cells co-cultured with unloaded erythrocytes (P
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Objective To explore the optimal method and condition for controlling release of vincristine (VCR) from VCR-loaded erythrocytes and enhancing the stability of carrier erythrocytes. Methods Erythrocytes were loaded into human erythrocytes using the modified hypotonic pre-swelling and isotonic resealing technique, and then with VCR, and they were treated with glutaraldehyde in concentration of 0.16% and 0.25% respectively. After storing at 4℃ for indicated time, the amount of VCR was determined with HPLC to compare their drug-release rate, and the degree of hemolysis in the supernatants was observed to compare the fragility of VCR-loaded erythrocytes treated with 0.16% glutaraldehyde with that with 0.25%, VCR-loaded erythrocytes treated with PBS, and VCR-loaded erythrocytes exposed to different hypotonic sodium chloride in different concentrations. Results The VCR-accumulated release rate increased in glutaraldehyde-treated VCR-loaded erythrocytes as well as untreated group during preservation. The VCR-accumulated release rate of 0.25% glutaraldehyde-treated group decreased by 71.67?4.20%, which was significantly slower than untreated VCR-loaded erythrocytes and 0.16% glutaraldehyde-treated group. After storing at 4℃ for 21 days, no significant changes at erythrocyte morphology was found in VCR-loaded erythrocytes treated with 0.25% glutaraldehyde, whereas untreated VCR-loaded erythrocytes and those treated with 0.16% glutaraldehyde can be stored only for 5d and 7.5d respectively. As compared to un-treated VCR-loaded erythrocytes, no significant change of osmotic fragility was seen in 0.25% glutaraldehyde-treated group, but significant increase in osmotic fragility was seen in 0.16% glutaraldehyde-treated group. Conclusion 0.25% glutaraldehyde could optimally block the membrane of VCR-loaded human erythrocytes and enhance their stability.
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Objective To study the relationship between innate immunity of lymphocytes and hu-moral and cell-mediated immunity in patients with psoriasis.Methods The rapid natural immune reaction on cancer cells was measured by lymphocytes isolated from fresh blood.Serum IgG,IgA,IgM were measured with rate-nephelometry in patients with psoriasis,and sandwich ELISA technique was applied to detect serum sIL-2R level in those patients.Results Rosette formation rate of lymphocytes adhering to cancer cells and serum immunoglobulin IgA level were significantly higher in patients with psoriasis than those in normal controls(P
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Objective To study CD35on erythrocytes and erythrocyte chemokine recepter(ECKR)in patients with psoriasis in order to explore red blood cell(RBC)innate immune function and its possible role in the pathogenesis of psoriasis.Methods The rapid natural immune reaction on cancer cells was measured with RBCs isolated from fresh blood.Expression of CD35on erythrocytes was detected by flow cy-tometry.The level of IL-8,which was bound to isolated erythrocytes,was measured by enzyme linked im-munosorbent assay(ELISA)in the supernatant after centrifugation,which represented ECKR binding activity.Results Rosette formation rates of RBC adhering to cancer cells and expression of CD35on RBCs were significantly higher in psoriatic patients than those in control group(P
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This paper discussed anti blood group antibody and red blood cell innate immunological function changes in gravidas with various blood groups. The 1∶64 positive rate of anti blood group antibody and innate immunological adhesive ratio of CR1 on red blood cell of gravida were assayed. Results showed that gravida with blood group AB didnot yield anti blood group antibody for antagonizing her husband blood group.However,the anti blood group antibody positive rate of gravida with blood group O was topmost(18.5%), which was also comparatable to group A (8.7%) and group B (3.0%)respectively with significant difference ( P
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Objective: To study the treatment of complicated and giant carotid body tumor. Methods: A giant carotid body tumor was dissected 2 years ago. The external carotid artery was embolized by means of catheterization before surgical intervention, and techniques of internal shunting and autogenous blood transfusion was used during the operation. Results: The tumor was resected completely without cerebral vessel disorders and major cephalic nerve injuries. Conclusion: The application of radial intervention of external carotid artery, shunting of internal carotid artery and autogenous blood transfusion are helpful to the treatment of giant and re operative carotid body tumor.
