Objective To improve HLA genotyping performance in clinical laboratory,the results of National Proficiency Testing（PT）for HLA-A/B/DRB1 low-resolution genotyping were collected and summarized from 2017 to 2020. Methods HLA genotyping results from 2017~2020 PT were analyzed and different kinds of errors were described and classified,and the possible reasons were also analyzed. Results The number of participant laboratories was increasing from 78 to 96 during 2017 and 2020. 12 096 HLA alleles were returned from the participants during the 4 years and total of 127 errors（1.05%）were found. These 127 errors were classified into two major types including 13 technical errors and 114 artificial errors. The proportion of laboratory which made mistakes was 2.56％（2/78）、4.88％（4/82）、8.75％（7/80） and 3.13％（3/96）in 2017,2018,2019 and 2020,respectively. Conclusion There were above 2.5% of laboratories with errors in HLA genotyping PT surveys past four years. More attentions should be greatly paid to clinical HLA genotyping test and the procedure of reporting.

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.

Objective To explore the predictive effect of early pregnancy placental growth factor （PlGF） combined with pregnancy associated protein A（PAPP-A）, mean arterial pressure（MAP）and uterine artery pulsatility index（UtPI）on preeclampsia,and to analyze the positive rate,incidence rate and efficiency of screening, so as to provide some data basis for the clinical application of preeclampsia screening in Anhui Province. Methods From 2019 to 2021, six hospitals in Anhui Province were organized to carry out a multi center research project to collect the clinical information of pregnant women who met the enrollment criteria within 11~13 ⁺⁶ gestational weeks. PlGF and PAPP-A were detected by time-resolved fluoroimmunoassay. The eclampsia risk assessment software was used to calculate the combined risk of multiple indicators in combination with maternal factors,PlGF,PAPP-A,MAP and UtPI,and to track the pregnancy development and pregnancy outcome of pregnant women. The clinical application effect of this screening model was analyzed by data statistics. Results A total of 9 400 pregnant cases were collected. There were 243 pregnant women with pregnancy related hypertension,including 128 cases of preeclampsia,6 cases of chronic hypertension with preeclampsia,101 cases of gestational hypertension,and 8 cases of pregnancy with chronic hypertension. The incidence rate of preeclampsia was about 1.43%. The cases of chronic hypertension with preeclampsia,gestational hypertension and pregnancy with chronic hypertension were excluded,and the remaining 9 285 pregnant women were included in this study. 128 cases of preeclampsia were taken as the study group,and the other 9 157 cases were taken as the control group. The results showed that PAPP-A and PlGF in the study group were lower than those in the control group （P<0.05）,and MAP and UtPI were higher than those in the control group（P<0.05）. A total of 6 139 samples with complete quadruple indicators （PlGF+PAPP-A+MAP+UtPI） were collected for separate statistical analysis. When the cut-off value was 1/137,the false positive rate of PlGF+PAPP-A+MAP+UtPI quadruple screening scheme was 15%,and the positive rates of PE<32 weeks, PE<34 weeks and PE<37 weeks were 1.16%,3.31% and 15.08% respectively,and the detection rates were 90%,80% and 63.64% respectively. Conclusion PlGF combined with PAPP-A,MAP and UtPI in early pregnancy is of high value in predicting early-onset preeclampsia.

Objective To investigate the effectiveness of reagents and assays suitable for screening tests for accidental antibodies to the red blood cell blood group system in daratumumab-treated patients. Methods Red blood group antibody screening cells were treated with 0.2 mol/L dithiothreitol（DTT） and 0.01 mol/L DTT,and the plasma of 13 patients treated with darettuzumab was accidentally screened for antibodies using the treated reagent red blood cells（microcolumn gel method）. The hemolysis degree,antigenicity of blood group antigen and red blood cell count were measured after treatment. Results Both concentrations of reagent red blood cells were uniformly detected. Negative results from the unexpected antibody screening test were observed in 11 patients,and unexpected antibodies（anti-E）in 2. Hemolysis occurred in 0.2 mol/L DTT treated reagent red blood cells on the 5th day of storage at 4℃,while there was no hemolysis within 31 days after treatment with 0.01 mol/L DTT,and red blood cell count was not statistically different from that of untreated reagent cells（P=0.59,0.40,0.78,all greater than 0.05）. The agglutination intensity of erythrocytes D,Fy ^a,Fy ^b,M,N,C,c,E,e,JK ^a,JK ^b,Le ^a,Le ^b,P1 and S antigens remained unchanged, but the agglutination intensity of K and k antigens was reduced. Conclusion After 0.01 mol/L DTT treatment,RBC blood group antibody screening cells could be used to detect unexpected antibodies in the RBC blood group system of patients receiving darettuzumab treatment,effectively eliminate the interference of darettuzumab and identify alloantibodies. Moreover,long-term storage can be achieved at 4℃,which does not necessitate temporary treatment of reagent red blood cells, thus saving the test time.

Objective To investigate the clinical efficacy of different doses of anthracycline idarubicin in the induction treatment of acute myeloid leukemia（AML）for adults aged less than 60. Methods A total of 265 young AML patients（14~59 years old）were diagnosed at our center from June 2015 to December 2019. We retrospectively reviewed 239 patients treated with the regimen of idarubicin combined with cytarabine（IA 3+7）,focusing on complete remission（CR）rate,recurrence and long-term survival in patients with different prognostic risk groups. Results After the first induction therapy,61（65.6%）and 103（70.6%） patients in the IDA 8 mg/m ² group and the IDA 10~12 mg/m ² group obtained morphological CR（P=0.475）,of which 26 patients（42.6%）in the IDA 8 mg/m ² group and 64 patients（62.1%）in the IDA 10~12 mg/m ² group had minimal residual disease（MRD）negativity. The 3-year cumulative incidence of relapse（CIR） was 45.1%（95% CI：34.1%~57.8%）in the IDA 8 mg/m ² group and 49.6% （95% CI：40.7%~59.3%） in the IDA 10~12 mg/m ² group（P=0.469）. The 3-year overall survival（OS）rate was lower in the IDA 8 mg/m ² group [34.6%（95% CI：24.9%~44.4%）] than in the IDA 10~12 mg/m ² group [46.6%（95% CI：38.2%~54.6%）] （P=0.038）. For intermediate-risk patients,the 3-year OS rate for patients in the IDA 8 mg/m ² group was 31.5%（95% CI：18.9%~45.0%）,which was significantly lower than that in the IDA 10~12 mg/m ² group [43.7%（95% CI：32.3%~54.6%）] （P=0.043）. The 3-year event-free survival（EFS） rate in the IDA 8 mg/m ² group and the IDA 10~12 mg/m ² group were 32.0% （95% CI：22.7%~41.6%）and 37.0%（95% CI：29.2%~44.8%）respectively（P=0.319）. For intermediate-risk patients, there was a lower trend of the 3-year EFS rate in the IDA 8 mg/m ² group [25.4%（95% CI：14.0%~38.4%）] than in the IDA 10~12 mg/m ² group [33.3%（95% CI：23.0%~44.0%）] （P=0.107）. There were no significant differences in the 3-year OS rate and 3-year EFS rate between the favourable risk group and the adverse risk group. Conclusion For young adult patients with AML,the induction therapy with IA (3+7) regimen based on idarubicin 10~12 mg•m ^-2•d ^-1×3 d could achieve higher immunological remission and better long-term survival,especially for patients with intermediate risk according to 2017 ELN risk stratification.

Objective We aimed to establish a relatively complete operation procedure for platelet storage using classical SSP+ platelet additive solution, and to compare the quality of preserved platelet concentrates in SSP+ versus in plasma. Methods Eighty-four blood （400 mL/person） were randomly collected from the general population in Chongqing. A buffy coats product derived from the whole blood donations of 6 same blood type donors were pooled to make a buffy coats platelet concentrate（n=14）. Each pool was divided into two groups, suspended in SSP+ and plasma, respectively. Platelet count, pH, MPV, platelet metabolism, hypotonic shock response（HSR）, CD62p expression and apoptosis indicator （AnnexinⅤ） were measured on Days 1, 5 and 7. Results There were no significant differences in platelet concentration, content, volume, MPV and pO ₂ during 7 days of storage between SSP+ group and plasma group （P>0.05）. However, the pH value in SSP+ group was significantly higher than that in plasma group （P<0.05）. The glucose consumption in SSP+ PAS group and the corresponding increase of lactic acid was significantly lower than that in plasma group （P<0.05）. With the decrease of pCO ₂, CO ₂ emission increased. The consumption of bicarbonate in additive SSP+ group was significantly lower than that in plasma group （P<0.05）. The HSR in SSP+ group was higher than that in plasma group. Both the CD62p expression and AnnexinⅤ were lower than that in plasma group. All products were performed the test for sterility on the 7th day of storage, and the results were negative. On the 7th day of storage, the pH and MPV in SSP+ group were still remained 6.94 ±0.11 and 9.89±0.52, respectively. Conclusion Platelets concentrates suspended in the classical SSP+ could metabolize normally within 7 days of storage. It has better in vitro parameters in platelet metabolism, platelet activation and apoptosis.

Objective To establish the surgical blood order schedule （SBOS） and explore the influencing factors of blood transfusion for orthopedic surgery patients. Methods We retrospectively enrolled 2 930 patients undergoing elective orthopaedic surgery in our hospital from January 2018 to December 2019,collected the application and actual usage of red blood cells（RBCs）,and analyzed the patients' blood transfusion rate,per capita RBCs transfusion,and whether there is a risk of major bleeding according to the type of surgery,to formulate SBOS for elective orthopaedic surgery. We used chi-square and Logistic regression methods to analyze the transfusion information and screened out the factors affecting perioperative transfusion. Results （1）The amount of RBCs application was 5 256 U by 2 930 patients of 15 surgical categories. The amount of RBCs transfusion was 538 U by 142 cases,with transfusion rate 4.85% and the per capita transfused RBCs amount was（3.93±2.63）U. The SBOS was divide into 3 categories：free Typing/Antibody-Screening （T/A-S）,only T/A-S and T/A-S with Crossmatch. （2）Multivariate Logistic regression analysis showed that the patient's age, preoperative hemoglobin level, and multiple injuries had an effect on perioperative RBCs transfusion（OR=0.145,3.058, 5.683,all P<0.05）. Conclusion The establishment of SBOS in orthopedic surgery provides a basis for better guidance of clinical blood transfusion. At the same time,the preoperative hemoglobin level,multiple injuries and age should be fully considered.

Objective To compare the anticoagulant effect of different concentrations of sodium citrate staged anticoagulation in hemodialysis patients. Methods 150 patients who received hemodialysis in the hospital from January 2019 to January 2021 were selected as the research subjects, and a random number table method was used to divide them into 5 groups with 30 cases in each group. In the 2.5% sodium citrate one-stage anticoagulation group and 4% sodium citrate one-stage anticoagulation group, 2.5% and 4% sodium citrate was pumped at the dialyzer; in the 2.5% sodium citrate staged anticoagulation group and 4% sodium citrate staged anticoagulation group, 2.5% and 4% sodium citrate was pumped at the dialyzer and venous kettle. At the end of treatment, the anticoagulant effect among the five groups was compared. The coagulation function indexes [thrombin time (TT), prothrombin time (PT), fibrinogen (FIB), activated partial thromboplastin time (APTT)], electrolyte indexes [bicarbonate (HCO ₃ ^-), calcium ion (Ca ²⁺), potassium ion (K ⁺), pH] and free calcium ion [arterial end (A), after filter (V)] were measured and compared before treatment (T ₀), on the 1 ^st h (T ₁), 2 ^nd h (T ₂), 3 ^rd h (T ₃) and 4 ^th h (T ₄) after treatment (iCa ²⁺). The dialysis adequacy (urea nitrogen clearance rate, urea clearance/volume) of patients among the five groups were compared. During treatment, the adverse reactions among the five groups were counted. Results The anticoagulation effect of 4% sodium citrate staged anticoagulation group was better than that of non heparin group, 2.5% sodium citrate one-stage anticoagulation group and 4% sodium citrate one-stage anticoagulation group, and the total effective rate of anticoagulation was higher than that of non heparin group, 2.5% sodium citrate one-stage anticoagulation group and 4% sodium citrate one-stage anticoagulation group, there were statistical significant differences (P<0.05); there was no statistical significant difference in the total effective rate of anticoagulation between 2.5% sodium citrate segmented anticoagulation group and 4% sodium citrate segmented anticoagulation group (P>0.05); there was no statistical difference in PT, TT, APTT and FIB at T ₀, T ₁, T ₂, T ₃ and T ₄ among the five groups (P>0.05); compared PT, TT, APTT and FIB among the five groups at T ₀ with those at T ₁, T ₂, T ₃ and T ₄, there were statistical significant differences (P<0.05); there was no statistical difference in K ⁺ and HCO ₃ ^- at T ₀ among the five groups (P>0.05); compared K ⁺, HCO ₃ ^-, Ca ²⁺, pH among the five groups at T ₀ with those at T ₁, T ₂, T ₃ and T ₄, there were statistical significant differences (P<0.05), 2.5% sodium citrate staged anticoagulation group was lower than non heparin group, 2.5% sodium citrate one-stage anticoagulation group, 4% sodium citrate one-stage anticoagulation group and 4% sodium citrate staged anticoagulation group (P<0.05); compared K ⁺, HCO ₃ ^- among the five groups at T ₀ with those at T ₁, T ₂, T ₃ and T ₄ (P<0.05); there was no significant difference in Ca ²⁺ and pH among the five groups at T ₀, T ₁, T ₂, T ₃ and T ₄ (P>0.05); there was no significant difference in A iCa ²⁺ and ViCa ²⁺ at T ₀, T ₁, T ₂, T ₃ and T ₄ among the five groups (P>0.05); compared A iCa ²⁺ and ViCa ²⁺ among 5 groups at T ₀ with those at T ₁, T ₂, T ₃ and T ₄, there were statistical significant differences (P<0.05); there was no statistical significant difference in dialysis adequacy (urea nitrogen clearance rate, urea clearance/volume) of patients among the five groups (P>0.05); there was no statistical significant difference in the incidence of adverse reactions among the five groups (P>0.05). Conclusion Pumping 2.5% and 4% sodium citrate in front of dialyzer and venous kettle has high anticoagulant effect in hemodialysis patients, of which pumping 2.5% sodium citrate at the dialyzer and venous kettle can effectively adjust the electrolyte level of patients, and has certain safety.

Objective To investigate the correction performance of platelet optical detection technology of Mindray BC6800 blood analyzer for EDTA-dependent pseudothrombocytopenia（EDTA-PTCP）. Methods Totally 53 patients with EDTA-PTCP were assigned into the experimental group and 25 without platelet aggregation into the control group. Within 20 minutes after the specimens were collected,the blood of EDTA-K ₂ anticoagulant was detected by CDR mode with Mindray BC6800,and the count values of PLT-I and PLT-O were obtained. Meanwhile,non anticoagulated venous blood was counted by artificial microscope and recorded as PLT-M. The three groups of data were compared and analyzed. In addition, according to the number of platelet aggregation particles under microscope,the experimental group was divided into 5 groups：PLT<6,PLT 6~10,PLT 11~15,PLT 16~20 and PLT>20. Taking PLT-M as the target value and CV≤12.5% as the judgment standard, we counted the qualification rate of PLT-O correction counting in each group and analyzed the unqualified specimens. Additionally,10 specimens with successful correction were randomly selected and placed for 40 minutes and 60 minutes respectively,and then the number of qualified correction of the instrument was counted, with the smear simultaneously examined by microscope. Results There was significant difference between PLT-I and PLT-M in the experimental group（P<0.05）,while PLT-O and PLT-M did not differ significantly（P=0.317）. No significant difference was observed between PLT-M and PLT-O or PLT-I in the control group. The qualification rates of PLT-O correction counting in 5 groups of specimens with different aggregate particle numbers were 88.7%,88.7%,81.8%,80% and 22.2%, respectively. Compared with the blood smears of corrected and qualified specimens,the aggregation of platelets in corrected but failed specimens was more dense and clumpy,and the boundary of platelets was blurred. After being placed for 40 and 60 minutes, the qualification rate of correction was 70% and 50%. The number of platelet aggregation particles in the newly added specimens with unqualified correction was >20. Conclusion The optical detection technology of BC6800 blood analyzer can be used to correct the specimen counting in patients with EDTA-PTCP, but the accuracy of corrected counting results should be determined by combining the approximate particle number of platelet aggregation under blood smear and the density of adhesion between platelets.

Objective To establish a prediction model of fetomaternal hemorrhage syndrome on machine learning algorithm, so as to assist clinicians in early detection,diagnosis and intervention of FMH. Methods This study was based on the data of pregnant women at 6~42 weeks of gestation who have received antenatal examinations in the Second Xiangya Hospital between June 2019 and December 2020. The recursive feature elimination algorithm was used to select key variables and developed eight kinds of machine learning algorithms and traditional regression model,and the model with the best performance was selected. Besides,the ten-fold cross-validation method was used to verify the model. Results The AUC of XGBoost model is 0.827,the AUC of test set is 0.808,and the accuracy is 0.76. The performance of XGBoost model is significantly better than that of traditional logistic regression model with AUC of 0.681 and other seven machine learning models. Conclusion In this study,a prediction model of FMH based on XGBoost algorithm was successfully constructed and its prediction performance is good.

Objective Develop a suitable in vitro model to observe the adhesion of platelets on the vessel wall under pathological vascular conditions（including stenosis,blood flow disturbance,venous valve damage,etc.）. Methods In this study,an improved parallel flow chamber device was designed to detect the adhesion of platelets under physiological and pathological conditions with different shear rates and different vessel diameters. At the same time,the aggregation of platelets that flow through the chamber but not adhered was measured. Results Peripheral blood with fluorescently labeled platelets flows through the collagen-coated flow chamber at a normal flow rate. Platelets adhered to the tube wall can be detected in about 5~8 minutes. The maximum platelet coverage area is about 60%; the inhibitor SZ-2 can significantly inhibit platelet adhesion. The same platelet adhesion area increases in the local stenosis lumen and the lumen with blood vessel branches; the platelet aggregation rate flowing through the local stenosis lumen is significantly higher than that of the platelets flowing through the normal lumen. Conclusion The improved parallel flow chamber can simulate the blood flow state under physiological and vascular pathological conditions,and has application value for researching platelet function and monitoring the efficacy of antithrombotic reagents.