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 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.

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 To observe whether neoantigen peptides sequenced from tumor patients can induce allo-donor sourced PBMC to be specific reactive T cell. Methods PBMC were separated from healthy donors' buffy coat, then monocytes were collected using adhesion method, remaining PBL was kept frozen in -80℃. GM-CSF and IL-4 were added to induce monocytes into dendritic cells every three days. At Day 7 mature dendritic cells were harvested and plated in 24 well-plates, then these plates were loaded with 16 neoantigen peptides as one peptide in each well, 13/16 peptides were designed by our team. Five hours later PBLs recovered from -80℃ were added into these plates to be co-cultured with these treated DC in ratio of 4~10∶1. No peptide wells (DC only) were negative controls, and PBL only wells were prepared for adding PHA later as positive control. IL-2 was replenished in these plates every three days. Peptides were pulsed twice more at Day 14 and Day 21 respectively. Supernatant of each well was collected at Day 22 to measure IFN-γ concentration using ELISA methods, remaining cells were analyzed using Flowcytometry to measure the percentages of CD3 ⁺IFN-γ ⁺ or CD8 ⁺IFN-γ ⁺Tcells. Results 10 of 13 peptides can induce more than 3 kinds of PBL to proliferate into reactive CD3 ⁺ or CD8 ⁺ T cells, similar increases were also found in IFN-γ concentration（r=0.66, 0.58 for CD3 ⁺, CD8 ⁺ respectively, P<0.05) in supernatant by ELISA method. Conclusions Our research data demonstrate these neoantigen peptides sequenced from tumor patients can induce allo-blood donors'PBMC proliferate to neoantigen specific reactive T cells.

Anemia leads to increased perioperative blood transfusion rate and blood transfusion volume, prolonged hospital stay in coronary artery bypass grafting patients, and is also an independent risk factor for postoperative complications and mortality.Patient blood management is a patient-centered, systematic, evidence-based approach to improve patient outcomes through the diagnosis and treatment of perioperative anemia and the protection of autologous blood.This paper reviews the research progress in the prevention and treatment of perioperative anemia in patients with coronary artery bypass grafting, which hopely improves the treatment plan and clinical outcome of patients with anemia after coronary artery bypass grafting.

Objective We evaluated the quality of leukocyte-reduced suspended red blood cells (SRBCs) with storage time by detecting hemolysis rate, bacterial growth, biochemical indicators and indexes of energy metabolism, membrane damage and oxidative stress at 4 ℃. Methods On the day of collection, 50 mL of whole blood was collected from 10 volunteers as test specimens. Leukoreduction filters were used for the removal of leukocytes and the plasma was removed as much as possible on the day of collection. We added erythrocyte preservation solution according to a ratio of preservative solution per 200 mL of whole blood and stored at 4℃ until the end of study. The hemolysis rate, ATP/2, 3-DPG, Na ⁺/K ⁺/GLU/LDH/pH, PS/CD47 and MDA/SOD were tested on day 0, 7, 14, 21, 28, 35 and 42 of storage. Bacterial growth was measured on day 0, 14, 28 and 42 of storage. Results The hemolysis rate progressively increased with storage, which increased from 0.03% to 0.4% at from day 0 to 42. There were statistically significant differences compared to day 0 (P<0.001), but none of them exceeded the national standard requirements. There was no bacterial growth. Na ⁺, GLU and pH decreased with increasing K ⁺ and LDH levels in supernatant. ATP began to decrease on Day 14, dropping to 36.4% on Day 21 and to 13.2% at the end. There were significant differences compared to day 0 (P<0.001). 2, 3-DPG began to decline on day 7, decreased to 57.5% on day 28 and 44.0% on day 42. At the end of storage, external exposure of membrane phosphatidylserine (PS) increased to 0.83%, and the mean fluorescence intensity of CD47 decreased to 60.8%. There was no change in MDA value during storage. Activities of SOD decreased on day 28, with statistical difference compared to Day 0 (P<0.001). Conclusion By evaluating the quality of leukocyte-reduced SRBCs stored at 4 ℃, we suggested that the optimal storage period of leukocyte-reduced SRBCs was within two weeks, because indexes of biochemical, energy metabolic and oxygen-carrying capacity were basically maintained in two weeks.

Objective Marvelous advancements have been made in cancer therapies to improve clinical outcomes over last few years. However, therapeutic resistance has always been a major difficulty in cancer therapy, with extremely complicated mechanisms remain elusive. N6-methyladenosine (m6A) RNA modification, a hotspot in epigenetics, has gained growing attention as a potential determinant of therapeutic resistance. As the most prevalent RNA modification, m6A is involved in every links of RNA metabolism, including RNA splicing, nuclear export, translation and stability. Three kinds of regulators, methyltransferase (writer), demethylase (eraser) and RNA binding proteins (reader), together orchestrate the dynamic and reversible process of m6A modification. Herein, we primarily reviewed the regulatory mechanisms of m6A in therapeutic resistance, including chemotherapy and targeted therapy. Then we discussed the clinical potential of m6A modification to overcome resistance and optimize cancer therapy. Additionally, we proposed prospects of m6A modification for future research.

Objective To investigate the mechanism by which evodiamine alleviates blood transfusion related acute lung injury (TRALI) in rats by regulating high mobility group protein Bl (HMGB1)/ toll like receptor 4 (TLR4)/nuclear transcription factor-κB (NF-κB) signaling pathway. Methods A total of 72 SD rats were randomly divided into the model group, low-dose evodiamine group, high-dose evodiamine group, no load group, high-dose evodiamine+HMGB1 overexpression group, and control group with 12 rats in each. Except for the control group, the TRALI model was prepared by intraperitoneal injection of lipopolysaccharide (LPS) combined with infusion of human plasma. After that, we analyzed the lung function indexes such as inspiratory resistance (Ri), minute ventilation (MV) and peak expiratory flow (PEF). Then, in each group we examined the lung wet-dry ratio of rats and the pathological changes of lung tissue by hematoxylin-eosin (HE) staining; the platelet and neutrophil aggregation in lung tissue by immunofluorescence staining; the total number of cells and the percentage of neutrophils in bronchoalveolar lavage fluid (BALF); the levels of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in BALF and serum by enzyme-linked immunosorbent assay (ELISA); as well as the expression of HMGB1/TLR4/NF-κB signaling pathway-related proteins in lung tissue by western blotting. Results Compared with the model group, the pulmonary function indexes MV and PEF in low-dose evodiamine group and high-dose evodiamine group were increased (P<0.05), while Ri, lung wet-dry ratio, lung histopathological score, platelet-neutrophil optical density in lung tissue, total number of cells in BALF, percentage of neutrophils, levels of pro-inflammatory factors TNF-α and IL-1β in BALF and serum, HMGB1 and TLR4 protein expression in lung tissue and p-NF-κB p65/NF-κB p65 were all decreased (P<0.05). Overexpression of HMGB1 attenuated the improvement of lung function by evodiamine in TRALI rats. Conclusion Evodiamine may inhibit the inflammation in TRALI rats by inhibiting the activation of HMGB1/TLR4/NF-κB signaling, thereby reducing its acute lung injury, and repairing its lung function.

Objective To investigate the value of the ratio of neutrophil to lymphocyte (NLR), monocyte to lymphocyte (MLR), platelet to lymphocyte (PLR), and systemic immune inflammatory index (SⅡ) in the diagnosis of lymph node metastasis of gastric cancer. Methods A total of 98 patients, which were screened for gastric cancer in our hospital from January 2017 to December 2019, were divided into gastric cancer group(61 cases) and gastric benign disease group(37 cases) by pathological diagnosis. According to the lymph node metastasis, gastric cancer group was further divided into lymph node metastasis group (35 cases) and non lymph node metastasis group (26 cases). NLR, MLR, PLR and SⅡ were calculated according to the results of preoperative blood routine test, and the efficiency of each index in the diagnosis of lymph node metastasis was analyzed. Results Among 61 cases of gastric cancer, 35 cases had lymph node metastasis, the metastasis rate was 57.38%. The levels of NLR, MLR, PLR and SⅡ in lymph node metastasis group were significantly higher than those in non lymph node metastasis group and gastric benign disease group(P<0.01). The area under ROC curve of NLR, MLR, PLR and SⅡ was 0.761, 0.708, 0.686 and 0.781, respectively. The critical values of NLR, MLR, PLR and SⅡ were 3.23, 0.29, 197.7 and 574.47, respectively. NLR, MLR, PLR and SⅡ were correlated with tumor differentiation, invasion, diameter and stage. Patients with lower differentiation, deeper invasion, larger diameter and higher stage had higher values of NLR, MLR, PLR and SⅡ (P<0.05). There was no significant difference in NLR, MLR, PLR and SⅡ (P>0.05) with tumor location. Conclusion NLR, MLR, PLR and SⅡ are related to the differentiation, invasion, diameter and stage of gastric cancer, which could be helpful for the diagnosis of lymph node metastasis of gastric cancer, and provide a reference for the choice of surgical methods and the scope of lymph node dissection.

Objective To analyze the efficacy of transfusion in patients with mismatched red blood cells caused by auto-antibodies, thus providing evidence for clinical safety and rational transfusion. Methods The clinical records and blood transfusion data of 35 patients with incompatible transfusion due to positive autoantibodies from May 2021 to June 2022 in our hospital were collected and statistically analyzed. Results A total of 60 transfusion applications had been made by 35 patients, 4 of which were rejected. Among 56 incompatible transfusions, 52 (92.9%) were found to be effective. There was no significant increase in TBIL after transfusion. No significant difference was observed in efficacy between transfusion of washed erythrocytes and that of non-washed erythrocytes (P>0.05). All patients had no transfusion reactions. Conclusion In case positive autoantibodies lead to blood mismatch, transfusion of ABO and Rh homologous red blood cells is safe and effective, whose primary agglutination strength is not higher than that of self-agglutination. It is advisable for blood transfusion departments to enhance doctor-patient communication to avert medical accidents; simultaneously, efforts should be made for more studies to prevent autoantibodies from affecting pre-transfusion tests.

Objective To explore the effect of platelet rich plasma (PRP) on traumatic synovitis of ankle joint. Methods From January 2021 to June 2022, 45 patients with traumatic synovitis of the ankle joint who were treated in the department of orthopedics and traumatology of a tertiary hospital were included. The patients were randomly divided into the observation group (20 cases, PRP treatment) and the control group (25 cases, traditional corticosteroid treatment) according to the random number table method. The ankle joint scores and clinical effects were compared between the two groups at before, 1, 3 and 6 months after treatment. Results The Baird-Jackson score and AOFAS score were higher (P<0.05) and VAS score was lower than before treatment in both groups after 1 month of treatment (P<0.05), and the control group was better than observation group (P<0.05). After 3 months of treatment, Baird-Jackson score and AOFAS score decreased (P<0.05) and VAS score increased (P<0.05) in the control group compared with one month after treatment. Baird-Jackson score and AOFAS score increased (P<0.05) and VAS score decreased (P<0.05) in the observation group compared with one month after treatment ( P<0.05), which was better than the control group (P<0.05). Time to first improvement in the control group was more common in 24 hours after treatment, and which was more common in 2 weeks and 1 month after treatment in the observation group (P<0.05). The total effective rate of the observation group was 95%, higher than that in the control group of 68% (P<0.05). Conclusion Traditional hormone therapy is fast-acting but short-sustained and prone to recurrence. PRP has better long-term effects and can improve patients' clinical symptoms for up to 6 months, which is worthy of clinical application.

Objective To investigate the effect of multiple apheresis platelet donation on some blood routine indicators and platelet correlation factors in plasma. Methods Sixty cases of multiple apheresis platelets donors who have donated platelets twenty times or more per year were listed as study group. 28 cases of first-time apheresis platelet donors were listed as control group. Before donating apheresis platelet, hemoglobin (HGB), hematocrit (HCT), platelet counts (PLT), cumulative number of blood donations were collected and analyzed. The level of thrombopoietin (TPO), platelet activation factor (PAF), von Willebrand factor (vWF), β-thromboglobulin (β-TG), platlet factor 4 (PF4) in plasma of two groups of blood donors before donation were measured by ELISA. Then those data were analyzed statistically. Results The mean age of the blood donors in study group was higher than that of the control group (P<0.05). By covariance analysis, we found that donors of different ages donated apheresis platelet 20 times or more per year, their peripheral blood HGB, HCT, PLT had no significant difference. Donors with different cumulative donation times donate 20 times or more per year, their peripheral blood PLT was no significant difference (P=0.142), but the donors with cumulative number of 51~100 times blood donation had higher HCT3, PLT3 than the other two groups (P=0.033, P=0.023). Different age and the cumulative number of blood donation had no significant influence in TPO, PAF, vWF, β-TG and PF4 (P>0.05). Conclusions Multiple apheresis platelets donations had no significant impact on peripheral HGB, HCT, and PLT. Also there was no obvious influence in platelet related factors.

Objective Correlation study between Common parameters of thromboelastography, Coagulatant factor, Vascular endothelial factor, Antithrombin, Plasminogen and traditional coagulation test. Methods Collecting any abnormal sample of the results of thromboelastography in our hospital for nearly one year as the research object, meanwhile, frozen the plasma of same sample. When a certain amount is collected, the test is performed. Results The results of multiple linear regression analysis showed that only PLG and Fig had a regression relationship with the parameters. Conclusions When the blood coagulation state of the patient is evaluated from the whole, the thromboelastogram is superior to other laboratory coagulation tests, but the possible results of the coagulation factor and coagulation cannot be inferred based on the R value, and there is no one-to-one correspondence between them.