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.

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.

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 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 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.

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 compare ELISA and colloidal gold immunoassay reagents in IgG and IgM antibody titer detection in plasma from the COVID-19 convalescent patients (COVID-19 CP). To determine the effect of methylene blue/photochemical virus inactivation on the titer of 2019-nCoV IgG. Methods Blood samples were collected from 15 COVID-19 convalescent patients discharged from hospital in different time, and tested according to the blood station technical operation rules (2019 Edition). Novel coronavirus pneumonia (2019-nCoV) RNAs were detected by nucleic acid testing. 3 IgG and 2 IgM ELISA kits,1 colloidal gold reagent were used to detect the antibody of IgM and IgG respectively, after diluted by origin, 20, 40, 80, 160 and 320 times. The titers of IgG antibody were detected in 7 plasma samples before and after the virus inactivation by methylene blue to evaluate the effect of virus inactivation on the titer of antibody. Results 15 samples of COVID-19 CP were qualified, and the 2019-nCoV nucleic acid test was negative. The ALT levels were high. There was a significant difference in the titer of IgG detected by 3 IgG and 2 IgM ELISA reagent kits from different manufacturers. Conclusion Domestic reagents can detect the IgG and IgM of 2019-nCoV, but the titer levels are different from the different reagents. It is necessary to further study the methodology and sensitivity of qualitative reagent and gradient dilution method for the detection of antibody titer. Methylene blue/photochemical virus inactivation has no significant effect on the titer of 2019-nCoV IgG

  Objective To investigate the quality and the influence factors of red blood cells in additive solution with reduced leukocytes to provide theoretical basis and reference methods of assessment indicators and reasonable dose for clinical red blood cell transfusion. Methods The blood volume, hemoglobin hematocrit and the hemolytic rate were analyzed at the end of storage in 100 bags of red blood cells in additive solution with leukocytes reduced. Result In the
group of 1.5 U red blood cells in additive solution with leukocytes reduced in 5 0 bags, the average blood volume was (198±22 ) ml/bag, and the qualified rate, compared with the theoretical value, was 78%. The hemoglobin concentration was (35.04±6.24 ) g, and the qualified rate was 92%. The hematocrit was (0.52±0.06 ) and, the qualified rate was 92%. The residual quantification of leukocyte was (2.15±0.80 )×1 0 ⁶/bag, and the qualified rate was 9 6%. The hemolytic rate at the end of storage was (0 .1 3±0 .1 2 )％, and the qualified rate was 1 0 0%. In the group of 2 .0 U red blood cells of 50 bags in additive solution with leukocytes reduced, the average blood volume was (272±26 ) ml/bag, and the qualified rate, compared with the theoretical value, was 75%. The hemoglobin concentration was (51.23±6.38 ) g, and the qualified rate was 96%. The hematocrit was (0.55±0.04 ), and the qualified rate was 100%. The residual quantification of leukocyte was (3.00±1.23 )×10 ⁶/bag，and the qualified rate was 94%. The hemolytic rate at the end of storage was (0.27±0.15 )％, and the qualified rate is 100%. Conclusion The quality indicators of blood products in both 1.5 U and 2.0 U of red blood cells in additive solution with leukocytes reduced met the standard of 2012 edition of "Technical Operating Procedures in Blood Station".

Objective To investigate the clinical effect of patients with Luminal A breast cancer treated with chemotherapy and endocrine therapy. Methods A total of 84 patients with Luminal A type breast cancer were observed in 2015 and 2017. The patients were randomly divided into control group （n=42） and observation group （n=42）. The control patients were treated with chemotherapy alone whereas those for observation were treated in combination with tamoxifen endocrine therapy. CD3 ⁺,CD4 ⁺,CD8 ⁺,and CD4 ⁺/CD8 ⁺cells were measured by flow cytometry pre- and post- treatment. Simultaneously,IFN-γ,TNF-α,IL-4,and IL-10 cytokines were detected. The survival time at different time points after treatments was compared for the clinical efficacy in the two groups. Results No significant difference was noted in the efficacy rate between the treatment group （64.29%） and the control （47.62%） （P>0.05）. The number of CD3 ⁺/CD4 ⁺CD8 ⁺ and CD4 ⁺/CD8 ⁺cells in the group of combination treatments was increased when compared with the control （P<0.05）. But the CD3 ⁺/CD8 ⁺cell counts as well as IL-10 in the combination group were lower than in the control （ P<0.05）. No significant difference,however,was found in survival rates of 1yr,3yr,and 5yr between the two groups （P>0.05）. Conclusion The combination of chemotherapy and tamoxifen endocrinotherapy in patients with Luminal A type breast cancer may help improve the immunity,inhibit inflammatory factors and prolong the survival of patients.

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.

The purpose of red blood cell transfusion is to increase hemoglobin and improve hypoxia.The reasons that affect transfusion efficacy can be divided into immune factors and non-immune factors.Immune factors include the changes of red blood cells surface molecules,content of metabolites,the influence of immune substances in blood recipients and so on.Non-immune factors include storage damage of red blood cells,disease status of blood recipients and drug effects.Physicians are more concerned with transfusion-related adverse reactions but less with poor red blood cells transfusion responses.Here,we review the potential reasons for the poor responses of red blood cell transfusion,in order to provide theoretical support and new ideas for research and clinical practice.

Objective To investigate the correlation between tumor markers CA72-4 and pepsin levels and colchicine treatment in gout patients. Methods From January 2016 to July 2017 in 64 cases of gout in Department of Endocrinology and Rheumatology in our hospital，the patients were divided into two group according to whether or not colchicine was taken in the first two weeks before admission. The patient's general data were collected including gender，age，duration of disease，and levels of CA72-4，and pepsin. In order to exclude gastrointestinal and other tumors，electronic gastroscopy，electronic colonoscopy，abdominal color Doppler ultrasound examination were carried out. Results ①64 patients had history of gout and no tumor diseases. ②There was no significant difference between the two groups in sex，age，course of disease，ESR，UA，CRP and PCT. ③CA72-4 group was significantly higher than non - taking group （P<0.05）. After two weeks' withdrawal，the level of CA72-4 decreased significantly，the difference was statistically significant （P<0.05）. ④The difference of PGI between the two groups during the period of medication and after withdrawal was statistically significant，but there was no statistical difference between PGII and PGI/PGII. Conclusion Colchicine treatment in patients with gout may be associated with elevated CA72-4. Check CA72-4 after stopping medication to exclude the impact of drugs on the test results; Colchicine treatment of gout and PGI have a certainly relevance.

Objective Analyze the distribution characteristics of weak positive results of irregular antibody screening tests to explore the significance in clinical transfusion practice. Methods 32 684 patients requiring transfusion in our hospital between July 2017 and June 2019 were selected and samples were screened for irregular antibodies using antiglobulin test. The positive samples were further tested for antibody identification using saline test and antiglobulin test tube method. The distribution characteristics and possible reasons of agglutination reaction≤1+ results were analyzed. Results Of the 71 cases with weak positive antibody screening results, 31 were male and 40 were female,but there was no statistically significant difference（P>0.05）. The weak positive rate of irregular antibodies was not related to age（P>0.05）. 43 cases had a blood transfusion, and 28 cases had no history of blood transfusion; 52 cases were specific alloantibodies; 10 cases could not determine antibody specificity; 9 cases were negative for panelcell-16 antibody identification. Conclusion More attention should be paid to weak positive irregular antibody screening results, and provide compatible blood products to avoid blood transfusion ineffectiveness and adverse events of hemolytic transfusion reactions.

Objective To compare the consistency of the results of hypersensitive c-reactive protein （hs-CRP） detected by three different instruments. Methods From march to July, 2018, 156 cases of patients with infection and fever who visited the outpatient department of Longhua district people's hospital in Shenzhen were selected. Using AU5800 as the reference instrument and VITROS5600 and 5390 as the comparison instruments, the consistency of hs-CRP test results of three different instruments was analyzed. Results In serum of 156 cases of hs-CRP reference instrument test results concentration was 93.64±79.74 mg/L, Mindray 5390 automatic blood corpuscle-hypersensitive c-reactive protein all-in-one test results was 87.98±76.57 mg/L, there was no statistically significant difference between the two （t=0.912 4, P>0.05）, while VITROS5600 dry piece of biochemical analyzer test results was 182.38±158.79 mg/L, significantly higher than the reference instrument, the difference was statistically significant （t=6.805 6, P<0.05）. The correlation coefficient （r） between hs-CRP concentration detected by the three instruments was 0.968 5. The correlation coefficient （r） between VITROS5600 and AU5800 was 0.953 0. Conclusion The results of hs-CRP detected by 5390 hemocyte-hypersensitive c-reactive protein integrator and AU5800 biochemical analyzer were consistent, while the results detected by VITROS5600 dry platebiochemical immune analyzer were obviously higher, but the hs-CRP detected by 3 different instruments showed good positive correlation.