Objectives To explore the possibility to utilize unrelated donors' cells as sources for adoptive T cell anti-tumor therapy, thus we need to identify HLA restriction for neoantigens and evaluate neoantigen induced reactive T cells' cytolysis ability. Methods 16 neoantigen peptides were used to induce 18 unrelated donors' (group 1) PBMC (peripheral blood mononuclear cells) into reactive T cells, and HLA typing was performed, the affinity between peptides' and HLA molecule was predicted by NetMHC database, then neoantigen with HLA-A2 restriction was selected to induce another group (group 2) of 17 unrelated donors' PBMC into reactive T cells, these cells would act as effectors, tumor cell line T2 cells or the tumor cells with the same source of this neoantigen would act as targets either, LDH release tests and RTCA (real time celluar analysis) were performed to evaluate the effectors' anti-tumor ability. Then these results were compared between HLA-A2+ and HLA-A2- samples. Results NetMHC database predicted neoantigen LM7 showed high affinity with HLA-A2+ antigens, 5/11 of HLA-A2+ samples can successfully be induced into reactive T cells, including 3/3 homozygous HLA-A2+ samples, while 2/7 of HLA-A2- samples with more reactive T cells. Induced T cells from A2+ samples showed higher percentages of tumor cells killed than those A2- samples(60.72±11.28 vs 47.2± 4.46, P=0.03). Conclusions Our study suggests prediction by NetMHC is more helpful in homozygous samples, neoantigens LM7 is confirmed as HLA-A2 restrictive, which can induce some HLA-A2+ PBMC into reactive T cells which can kill tumor cells more efficiently, unrelated donors should be screened not only by HLA-typing but also cell function tested, thus induced reactive T cells can take the roles as the source for adoptive anti-tumor T cells therapy.

Objective To retrospectively analyze the influencing factors of arachidonic acid （AA） and adenosine diphosphate （ADP） inhibition rates and the efficacy of different antiplatelet drugs. Methods A total of 114 patients with platelet inhibition rates detected in the hospital were selected from March 2015 to December 2018 . The inhibition rates of AA and ADP were divided into several groups according to the antiplatelet treatment strategies. Results The ADP inhibition rate,rather than the AA inhibition rate（P>0.05）,was negatively correlated with age （P<0.01） and positively with triglyceride （TG） test results （P<0.01）.Significant differences of ADP inhibition rates between groups of aspirin and aspirin+Greg los（P< 0.01） and between groups of clopidogrel and aspirin+Greg los（P<0.05） were noted.Similarly,remarkable differences of AA inhibition rates between groups of aspirin and clopidogrel（P<0.01）,clopidogrel and aspirin+Greg los（P<0.01）,and clopidogrel and aspirin+clopidogrel（P<0.05） were seen. Conclusion A comprehensive analysis of age and TG level should be taken into account in terms of the ADP inhibitory rate detection.Greg los treatment is effective based on the ADP inhibition rate measurement while aspirin therapy is promising based on AA inhibition.

Objectives To assess the recover cells from Leukocyte reduction filters (LRFs) and their related factors, this study analyzed the number of natural killer (NK) cells and T cells from LRFs and whole blood (WB). The correlation between the number of these cells and age, sex and other factors was further analyzed. Methods WB and LRFs (400 mL) were collected from 51 repeat blood donors in Shenzhen Blood Center from July 2023 to June 2024. White blood cells (WBCs) were obtained by washing LRFs with PBS buffer. The cells from WB and LRFs were counted by blood cell analyzer. Mononuclear cells were isolated by density gradient centrifugation. The proportions of CD3 ^-CD56 ⁺NK, CD3 ⁺CD4 ⁺T and CD3 ⁺CD8 ⁺T cells were detected by flow cytometry. Statistical analysis was performed using GraphPad Prism v.9.0 software. Results In a certain flush volume, the number of total lymphocytes from LRFs increased with the increase of flush volume, but the cell density decreased with the increase of flush solution. There was no significant difference in the total number of lymphocytes from LRFs among donors of different ages and genders ( P>0.05). According to the results of cell number and cell ratio, the number of CD3 ^-CD56 ⁺NK cells from LRFs was different in age groups ( P<0.05), but no significant difference was found in gender groups ( P>0.05). The number of CD3 ^-CD56 ⁺NK cells from WB in male donor were significantly higher than that in female donor ( P>0.05). Whether from LRFs or WB, there was no significant difference in the number of CD3 ⁺CD4 ⁺T cells and CD3+CD8+T cells in different age groups and gender groups ( P>0.05). Conclusion In a certain flush volume, the total number of lymphocytes from LRFs was correlated with the volume of the flush solution. The number of CD3 ⁺CD4 ⁺T cells and CD3 ⁺CD8 ⁺T cells were not significantly correlated with the age and gender of blood donor, while the number of CD3 ^-CD56 ⁺NK cells was correlated with the age of blood donor.

Objectives To evaluate the effect of virus reduction in plasma using LED white light. Methods Methylene blue was added to plasma at a final concentration of 1 µM. The activity of coagulation factors in plasma and the inactivation effect on Sindbis virus were compared by different treatment methods (10 cm/5 cm spacing, unilateral/bilateral irradiation), different light intensities (30 000 lx, 35 000 lx, 40 000 lx, 45 000 lx, and 50 000 lx) and different time points (10, 20, and 30 min) using LED white light as the light source. Results Increase time of irradiation led to increase of plasma temperature under 50 000 lx light intensity. The extent of increase was found to be greater at 5 cm than at 10 cm spacing, and greater with bilateral than unilateral irradiation. The 5 cm/10 cm spacing and unilateral/bilateral irradiation had no significant differences in their effect on coagulation factor Ⅷ and fibrinogen (FIB) activity. The 10 cm bilateral spacing was selected as the device parameter for photochemical treatment. Plasma was irradiated at intensities of 30 000, 35 000, 40 000, 45 000, and 50 000 lx, respectively. The results showed Sindbis virus in plasma was inactivated after 5 minutes of irradiation at different light intensities, with titer reduction >4 LogTCID ₅₀/0.1 mL. Factor Ⅷ and FIB activities were retained at the level of 70% and 60%, respectively after 20 minutes of irradiation. Correlation analysis revealed that there was no correlation between different light intensities and loss of factor Ⅷ activity. However, the effect of light intensities on FIB activity was more pronounced within 20 minutes of irradiation. A significant decline in the FIB activity was observed with increasing light intensities, although no significant difference was found after 30 minutes of irradiation. The irradiation time was found to be significantly correlated with the loss activity of coagulation factors, regardless of the light intensities. Conclusion LED white light can act as a new light source for methylene blue virus reduction. Under the right conditions it reduced viruses effectively and preserves the activity of the coagulation factors in plasma.

Objectives To study the clinical efficacy and safety of ABO incompatible platelet transfusion in emergency situations. Methods 271 cases of ABO incompatible platelet transfusion in our hospital from January 2020 to January 2024 were collected, and 128 cases were enrolled which were divided into bidirectional mismatch group (10 cases), major mismatch group (56 cases), and minor mismatch group (62 cases). All 128 patients had ABO homologous platelet transfusion history before and within 10 days after ABO incompatible platelet transfusion. The incompatible transfusion was set as the ABO blood group incompatible experimental group (subgroup B), the previous ABO homozygous platelet transfusion was set as the ABO homozygous control group (subgroup A), and the subsequent ABO homozygous platelet transfusion was set as the ABO homozygous experimental group (subgroup C). Platelet count change after platelet transfusion (△PLT) and 24 h platelet corrected count Increment (CCI) were used to evaluate the clinical effect of platelet transfusion, and the safety of platelet transfusion was assessed by the presence or absence of hemolytic transfusion reactions. Results In the bidirectional mismatch group, there was no significant difference in △PLT and CCI among subgroups A, B and C ( P>0.05). In the major mismatch group, the △PLT and CCI of subgroup B were significantly lower than those of subgroups A and C ( P<0.05), but there was no significant difference between subgroup A and C ( P>0.05). In the minor mismatch group, there was no significant difference in △PLT and CCI among subgroups A, B and C ( P>0.05). There was no acute hemolytic transfusion reactions in any of the ABO incompatibility infusion. Conclusion The clinical effect of ABO major mismatch platelet transfusion is obviously weaker than that of ABO compatible platelet transfusion, but did not affect the outcome of subsequent ABO homozygous platelet transfusion. The clinical effect of ABO minor mismatch platelet transfusion is similar to that of ABO compatible platelet transfusion, but could potentially affect the outcome of subsequent ABO homozygous platelet transfusions. No significant hemolytic transfusion reaction was found by ABO incompatible platelet transfusion.

Objectives To clarify whether NLRP3 inflammasome can modulate macrophage phagocytic capacity towards red blood cells (RBCs) and regulate macrophage subtype polarization. Methods THP-1 cells with low expression of NLRP3 (ID3 THP-1), cells transfected with an empty vector (shNC THP-1), and wild-type THP-1 cells were used as macrophage models. These cells were incubated with untreated RBCs, water bath-aged RBCs, and IgG-opsonized RBCs, respectively. Flow cytometry was used to detect the phagocytic rate of THP-1 cells to different RBC types and to measure the expression of M1 subtype markers (CD16 and CD86), and M2 subtype markers (CD163 and CD206) on THP-1 cells. Results ID3 THP-1 with reduced NLRP3 expression exhibited significantly downregulated phagocytic capacity towards RBCs. Aged RBCs induced the differentiation of THP-1 cells into the M1 subclass while inhibiting their differentiation into the M2 subclass. Decreased expression of the NLRP3 inflammasome in THP-1 cells led to a downregulation of their ability to differentiate into the M1 subclass following RBC phagocytosis, accompanied by their enhanced capacity to differentiate into the M2 subclass. Conclusion NLRP3 inflammasome can serve as a pivotal regulatory target, governing macrophage phagocytosis of RBCs and their subsequent subclass differentiation.

Objectives To investigate the effects of 4 ℃ cold-stored platelets (cold storage platelets, CSP) transfusion on acute kidney injury caused by hemorrhagic shock/resuscitation (HS/R) in rats. Methods 20 healthy male SD rats were randomly divided into 4 groups: sham operation group (blank group), HS/R model group (model group), CSP transfusion group, room temperature-stored platelets (RTP) kept at 22 ℃ transfusion group (RTP transfusion group). The rats were executed 24 hours after volume resuscitation, and blood and kidney tissue specimens were collected. Serum concentrations of creatinine (Scr), urea nitrogen (BUN) , cystatin C (CysC) and interleukin-1β (IL-1β) and interleukin-18 (IL-18) in renal tissue were detected in each group. The expression levels of neutrophil gelatinase-associated lipid carrier protein (NGAL) and renal injury cause-1 (KIM-1) in renal tissue were detected by Western Blot method, and the pathological changes of renal tissue were observed by HE staining. Results ①There were no significant differences in baseline body mass, MAP, SCr, BUN and other indexs among four groups ( P>0.05). ②24 h after resuscitation, the levels of SCr ( P<0.05), BUN ( P<0.01), CysC ( P<0.01) were higher in the model group than in the other three groups; the levels of BUN ( P<0.01), SCr ( P=0.037), CysC ( P=0.020) in the CSP infusion group were lower than those in the RTP infusion group. ③Compared with the model group, serum IL-1β and IL-18 levels were significantly lower in both CSP infusion group and RTP infusion group. Compared with the RTP infusion group, serum IL-1β levels were reduced in the CSP infusion group ( P=0.041). Compared with model group, NGAL protein expression was significantly decreased in CSP infusion group ( P<0.01), and KIM-1 protein expression was significantly decreased in both CSP infusion group and RTP infusion group ( P<0.01). Renal histopathological results showed that CSP transfusion could reduce the degree of renal cell degeneration, necrosis and inflammatory cell infiltration. Conclusion Compared with RTP, CSP transfusion could reduce HS/R acute kidney injury in SD rats.

Objective To explore the solution of interference in cross matching of patients with multiple myeloma treated with CD38 monoclonal antibody and to formulate its transfusion strategy. Methods Multiple myeloma patients treated with the CD38 monoclonal antibody Daratumab used dithiothreitol (DTT) before and after treatment with antibody screening cells,spectral cells,and donor red blood cells.Unexpected antibody screening,identification and cross-matching were performed by microcolumn Gel method and polybrene method respectively. Results After chemotherapy with CD38 monoclonal antibody,ABO blood groups were consistent with forward and reverse typing in 5 patients.The microcolumn Gel method was positive for antibody screening,identification and forward cross-matching,while the polybrene method was negative.Unexpected antibody screening cells,spectrum cells and donor red cells were treated with 0.2mol/L DTT.The microcolumn Gel method was negative in antibody screening,identification and forward typing.After blood transfusion,the patients' hemoglobin increased significantly,but the serum bilirubin did not change obviously. Conclusion In patients with multiple myeloma treated with CD38 monoclonal antibody,red blood cells treated with 0.2mol/L DTT can be used for antibody screening and cross matching by microcolumn Gel,or direct polybrene,which can be equally effective.

Whole blood was the earliest blood product ever used, however, with the advent of component transfusion treatments, whole blood all but disappeared from blood bank lists in the 1970 s. In recent years, based upon the successful military experience, the use of whole blood to resuscitate patients with hemorrhagic trauma has again attracted the attention of the blood transfusion circle,and it has gradually been introduced from the military field to the civilian treatment settings.Based on the guidelines for whole blood transfusion at home and abroad and related published studies, this paper reviews the classification of whole blood, history of whole blood use, whole blood storage and platelet protection, removal of whole blood white blood cells, inactivation of whole blood pathogens, and urgent problems to be solved in scientific and accurate transfusion of whole blood, and looks forward to its future development direction, providing reference for the introduction of whole blood more widely, and the development of blood transfusion medicine.

Objective To investigate the changes of serum calcium concentration（iCa） before and after massive transfusion，and discuss the influencing factors of the occurrence of hypocalcemia. Methods 75 cases of patients receiving massive blood transfusion in Xiangya Hospital from May 1，2015 to Apr 30，2016 were selected，collected serum calcium concentration of patients before and after massive transfusion，and analyzed changes of serum AG during massive transfusion，researched on the factors affecting the incidence of hypocalcemia. Results Student t test between groups showed that the serum calcium concentration（iCa） was significantly lower than that before the blood transfusion （P<0.05），iCa after the blood transfusion 24 hours rise again，but it was still lower than that before blood transfusion （P<0.05）. Logistic regression analysis showed that the amount of blood product input and AG after blood transfusion （blood acidity after transfusion） are important factors leading to the occurrence of hypocalcemia after massive transfusion，AG after blood transfusion is main influencing factor of the severity of hypocalcemia（P< 0.05）. Conclusion Hypocalcemia has direct relationships with the amount of blood products input （P＜0.05），while patients receiving massive transfusion have higher probability to suffer from hypocalcemia. Monitoring the change of iCa and blood pH closely during treatment can reduce the occurrence of hypocalcemia and other complications after massive transfusion.

Objective To investigate whether free heme released in hemolysis could directly damage hepatocytes and disrupt liver function. Methods The mice model of hemolytic transfusion reaction was established, and ALT, AST and other biochemical indexes were detected to analyze the liver function. In vitro, the LO2 cells were stimulated with different concentrations of lysed supernatant of red blood cells and heme respectively. Then the levels of ALT and AST were detected to analyze the liver function. Cell viability was observed by flow cytometry, and cell morphology and skeleton structure were observed by immunofluorescence. Results Abnormal liver function was observed on the mice model of hemolytic transfusion reaction. The results of in vitro experiments showed that LO2 cell viability decreased and the proportion of dead cells increased along with the incremental concentration of free heme. The biochemical indexes such as ALT and AST were aggravated significantly. And free heme could directly destroy the cytoskeleton of LO2 cells. Conclusion Free heme can directly destroy the cell structure of hepatocytes, inhibit cell vitality, induce cell death and abnormal liver function. The degree of damage is positively correlated with the concentration of free heme.

Objective To investigate the level and clinical value of autoantibodies in patients with malignant pulmonary nodules. Methods Seventy-one patients with asymptomatic pulmonary nodules admitted to our hospital from November 2018 to January 2019 were enrolled, including 58 malignant pulmonary nodules and 13 benign pulmonary nodules. And ten healthy people as control group.The levels of p53, PGP9.5, SOX2, GAGE7, GBU4-5, MAGEA1 and CAGE in all specimens were detected by ELISA kit. The ROC curve was used to analyze the diagnostic value of autoantibodies for malignant pulmonary nodules. Results Compared with the benign pulmonary nodules group and the normal control group, the level of autoantibodies in the malignant nodules of the lungs was significantly increased (P<0.05);ROC curve analysis showed that seven autoantibodies were important for the differential diagnosis of benign and malignant pulmonary nodules (AUC>0.5, P<0.05), and seven autoantibodies combined detection could improve the differential diagnosis efficiency (AUC=0.822);The positive detection rate of lung malignant nodules was 81.7% by LDCT and was increased to 94.0% combining with autoantibodies detection (P<0.05). Conclusion Serum autoantibody detection has potentialclinical value in the differential diagnosis of benign and malignant pulmonary nodules.

The treatment of chronic non-healing wounds remain challenging in terms of complexity. Although platelet rich plasma (PRP) therapy has been widely proven effective, the traditional method of in vitro activation of PRP leads to the rapid release of all growth factors. Due to the separation of colloid and supernatant after activation, the factor-rich supernatant is easy to flow and lose, and it is difficult to form a stable structure, which affects its therapeutic effect. To overcome this problem, researchers in recent years have explored hydrogels as carriers for PRP to improve the shortcomings of traditional PRP treatment. This article reviews the latest research on hydrogel-loaded PRP for the treatment of chronic non-healing wounds and provides a reference for optimal treatment.

Objective The invalid results and listsmay influence thetimely sending ofthe nucleic acid test reports. This work aimed to explore the reason for the invalid results and to optimize the corresponding operations for safety of blood transfusion. Methods Blood samples were collected from the voluntarydonors in Anyang from Jun 2016 to may 2017and DNAs were extracted withHaoyuan ChiTas BSS1200 automatic nucleic acid separator and subsequently amplified with ABI7500 Fluorescent Amplifier. The invalid results and lists were analyzed. Results A total of 88739 samples were tested for DNAs and 0.06% results and 1.04% lists were found to be invalid. Further analyses of the invalid results showed that it is not the batche's number of reagent but the insufficient amounts of samples as well as personal operation errorsor laboratory contaminationsthat causedthe invalid amplification of internal standard. Conclusion The invalid results and lists can be minimized by enhancing the training of the staff who are involved in nucleic acid tests,improving operating procedures,and avoiding laboratory contaminations by daily cleaning and disinfecting.

Objective To investigate the effects of platelets with different storage days and quantities on the proliferation of liver cancer cells. Methods Twelve type O platelet braids were collected from Nanning Central Blood Station and sent to the First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine. According to the storage days, they were divided into fresh PLT group and old PLT group, of which the fresh PLT group was platelets stored for 1 day. In the old PLT group, platelets stored for 4 days were used. The two groups of platelets were co-cultured with human liver cancer cell Huh-7. The confluence degree of Huh-7 cells in each group within 48 hours was observed by scratch test, and the invasion ability of Huh-7 cells in each group within 24 hours was observed by transwell test. Results The results of scratch test showed that the proliferation ability of Huh-7 cells was significantly enhanced after co-culture with platelets. When the same amount of PLT was added, old PLT could promote the proliferation of Huh-7 cells more strongly. When PLT with the same storage days was added, PLT with a higher number generally showed a relatively stronger ability to stimulate cell proliferation. The results of transwell experiment were similar to the results of scratch. When the number of PLT was the same, the number of Huh-7 cells invaded by old PLT group was more than that of fresh PLT group, and the difference was statistically significant (P<0.05). The more PLT was added into Huh-7 cells, the more invasive cells occurred in the fresh PLT group and the old PLT group, and the difference was statistically significant (P<0.05). Conclusion The ability of platelets to promote cell proliferation and invasion is positively correlated with the storage time of platelets and the number of platelets. The application rules of platelet products need to be analyzed according to the specific clinical conditions. In the treatment of patients with neoplastic diseases, the application of old platelets should be avoided as far as possible, and the transfusion of platelets should be minimized when platelets have to be used, so as to achieve the purpose of slowing down the proliferation and metastasis of tumor cells.

Objective To know the frequency of HLA antibodies in donors in Shanghai area, to provide basic data for the research on TRALI. Methods Samples of blood donors from October 2020 to April 2021 were randomly selected to detect HLA-specific antibodies by flow cytometry and microbead method, and the incidence of HLA antibodies was statistically analyzed. Results In 9 797 serum samples of blood donors, 1 715 (17.51%) were positive for HLA antibodies, among which the frequency of HLA antibodies in males and females was 4.81% and 28.08%. Excluding the history of blood transfusion, the frequency of HLA antibodies in women with no pregnancy history and women with pregnancy history was 12.10% and 36.62%, and comparative analysis the group of once, twice and three or more times pregnancies donors population, which frequency of HLA antibodies was 29.97%, 40.70% and 44.80%. The frequency of HLA antibodies in female donors with multiple pregnancy history was significantly higher than that in single pregnancy ( P<0.05). Conclusion Consultation of pregnancy history and HLA antibodies detection of female blood donors in blood stations can prevent from the occurrence of TRALI.

Objective To evaluate the differential biological effects of human serum albumin (HSA) derived from different sources on vascular endothelial cells. Methods Human umbilical vein endothelial cells (HUVEC) were serum-starved and then exposed to HSA from varied sources. Cellular responses were assessed using the CCK-8 assay for proliferation, Annexin V-FITC kit for apoptosis, and flow cytometry for cell cycle analysis. The proliferative, apoptotic and cell cycle effects of HSA from different sources were statistically compared. Additionally, the impact of HSA on HUVEC cell migration and tube formation was assessed via scratch assays and tube formation experiments. Results Yeast-derived rHSA1 did not affect HUVEC proliferation ( P=0.49, q=1.601), while other HSA sources significantly promoted it ( P<0.001, F=10.84). All HSA treatment groups exhibited an inhibitory effect on HUVEC apoptosis ( P<0.001), with no statistically significant differences in the proportions of live cells ( P=0.07, F=2.415), apoptotic cells ( P=0.2, F=1.624), and dead cells ( P=0.28, F=1.376) across treatment groups. Compared to the serum-free control group, except for the pHSA2 and pHSA6 treatment groups, which showed a significant decrease in G0/G1 phase cell proportion ( P<0.001) and a significant increase in G2/M phase cell proportion ( P<0.01), the proportions of cells in other cell cycle phases did not show significant changes in the HSA treatment groups. In the scratch assay, in contrast to the serum-free control group, the pHSA(1) and pHSA(2) groups exhibited a higher degree of wound healing at 12 h ( P<0.001), 24 h ( P<0.001), and 36 h ( P<0.001, P=0.002); however, no significant differences were observed in the rHSA(1) and rHSA(2) groups compared to the control group. Likewise, in the tube formation assay, the pHSA(1) and pHSA(2) groups significantly enhanced node formation ( P=0.001, P=0.005), tubular branching ( P<0.001), and mesh structure development ( P<0.001), whereas the rHSA(1) and rHSA(2)groups showed no such enhancements. Conclusion HSA from different sources significantly inhibits endothelial cell apoptosis under serum-free conditions, but exerts varying effects on proliferation, cell cycle regulation, cell migration, and tube formation of vascular endothelial cells.

Non-transfusional hemotherapy should be mainly carried out by the Transfusion Department, which basis is the apheresis technology by centrifugation. The plasmapheresis can be realized in two ways: centrifugal and membrane filtration, each of which has its own technical characteristics. Apheresis/blood purification based on centrifugation shows the advantages of higher plasma separation efficiency, shorter treatment time, less platelet loss, less destruction of red blood cells, and the use of citrate anticoagulation for non-continuous clinical treatment of critically ill patients. Secondary columns suitable for centrifugal technology can realize immunoadsorption, artificial liver support system, centrifugation-filtration plasmapheresis and inflammatory factor adsorption. Using increasingly sophisticated secondary column technology should be a useful supplement to traditional TPE.