Objective:To establish a practical patient-based internal quality control method for valproic acid drug concentration monitoring.Methods:Observational Study. A PBRTQC model based on the exponentially weighted moving average (EWMA) method was established using Python. All results of a total of 28, 757 valproic acid concentration data from February 1, 2023 to January 31, 2024 were collected and split into training set and validation set at a ratio of 80% and 20% respectively. The truncation limit (TL) was optimized by using the winsorized mean method and the trimmed mean method. Different weighting coefficients λ were set. Different TL and different λ were combined with the EWMA algorithm into different patient-based real-time quality control (PBRTQC) models. The optimized models were verified by introducing simulated constant errors (CE) and proportional errors (PE) respectively. The false positive alarm rate (FAR) was used to evaluate specificity, and the average number of patients before error detection (ANPed) was used to evaluate sensitivity. According to the daily test volume and quality target requirements, we comprehensively judged whether the performance evaluation indicators of FAR and ANPed meet the laboratory requirements. Bias detection curve was used for determination of the best model.Results:The parameters of the best PBRTQC model for valproic acid drug concentration monitoring are: trimmed mean method with 1.5 standard deviations (i.e., truncating data outside 1.5 standard deviations of the data mean), λ=0.01. The performance verification result shows that ANPed of CE and PE of this model are both less than 100. The comparison between the EQA results and the EWMA results show that the EWMA method results are comparable to the EQA results.Conclusion:A PBRTQC model for the valproic acid drug concentration monitoring project based on the EWMA method has been successfully established. It is comparable with both IQC and EQA results, which means PBRTQC may be used as a supplement to the quality control of daily quality control products.

Objective:To verify the performance of the immunoturbidimetric assay(ITMA)kit of detecting oxidized low-density lipoprotein(ox-LDL).Methods:In accordance with the requirements of China National Accreditation Service for Conformity Assessment[(CNAS)-GL037:2019]and the National Health Industry Standards,the precision,trueness,linear range and reportable range of ITMA test kit for ox-LDL were verified on AU5800 fully automatic biochemical analyzer,which were compared with enzyme linked immunosorbent assay(ELISA)of Mercodia comparison reagents for ox-LDL through experiment of methodological comparison.Results:The coefficients of variation(CV)values of within-run and between-run precisions of the two concentration levels of quality control materials in the ITMA reagent kit for ox-LDL were respectively(3.74%,3.36%)and(4.75%,5.49%).The trueness bias of the two concentration levels of standards were respectively 0.46%and 1.78%,and the linear range was(15-120 U/L),which verification regression coefficient was R2=0.999 5 with a slope of 0.989.The maximum dilution multiple was 4 times,and the upper limitation of the clinically reportable range was 240.00 U/L.The manufacturer-provided biological reference interval was 0-64.49 U/L,and the coincidence rate was 100%.The results of hemoglobin,bilirubin,triglycerides,high density lipoprotein(HDL),low density lipoprotein(LDL)and rheumatoid factor were respectively 10 mg/ml,342 μmol/L,10.0 mg/ml,100 mg/dl,200 mg/dl,<200 IU/ml.The stationary duration was there was not influence within 30 h after opened the kit.The correlation between ITMA and ELISA detection methods was excellent(R2=0.9766).The positive and negative coincidence rates of clinical identification were respectively 92.3%and 96.5%.Conclusion:The performances of ox-LDL ITMA test kit based on fully automatic biochemical analyzer can all meet the requirement of clinical application,which shows excellent consistency with the results of ELISA method.

Objective:To verify the performance of the immunoturbidimetric assay(ITMA)kit of detecting oxidized low-density lipoprotein(ox-LDL).Methods:In accordance with the requirements of China National Accreditation Service for Conformity Assessment[(CNAS)-GL037:2019]and the National Health Industry Standards,the precision,trueness,linear range and reportable range of ITMA test kit for ox-LDL were verified on AU5800 fully automatic biochemical analyzer,which were compared with enzyme linked immunosorbent assay(ELISA)of Mercodia comparison reagents for ox-LDL through experiment of methodological comparison.Results:The coefficients of variation(CV)values of within-run and between-run precisions of the two concentration levels of quality control materials in the ITMA reagent kit for ox-LDL were respectively(3.74%,3.36%)and(4.75%,5.49%).The trueness bias of the two concentration levels of standards were respectively 0.46%and 1.78%,and the linear range was(15-120 U/L),which verification regression coefficient was R2=0.999 5 with a slope of 0.989.The maximum dilution multiple was 4 times,and the upper limitation of the clinically reportable range was 240.00 U/L.The manufacturer-provided biological reference interval was 0-64.49 U/L,and the coincidence rate was 100%.The results of hemoglobin,bilirubin,triglycerides,high density lipoprotein(HDL),low density lipoprotein(LDL)and rheumatoid factor were respectively 10 mg/ml,342 μmol/L,10.0 mg/ml,100 mg/dl,200 mg/dl,<200 IU/ml.The stationary duration was there was not influence within 30 h after opened the kit.The correlation between ITMA and ELISA detection methods was excellent(R2=0.9766).The positive and negative coincidence rates of clinical identification were respectively 92.3%and 96.5%.Conclusion:The performances of ox-LDL ITMA test kit based on fully automatic biochemical analyzer can all meet the requirement of clinical application,which shows excellent consistency with the results of ELISA method.

Objective:To establish a practical patient-based internal quality control method for valproic acid drug concentration monitoring.Methods:Observational Study. A PBRTQC model based on the exponentially weighted moving average (EWMA) method was established using Python. All results of a total of 28, 757 valproic acid concentration data from February 1, 2023 to January 31, 2024 were collected and split into training set and validation set at a ratio of 80% and 20% respectively. The truncation limit (TL) was optimized by using the winsorized mean method and the trimmed mean method. Different weighting coefficients λ were set. Different TL and different λ were combined with the EWMA algorithm into different patient-based real-time quality control (PBRTQC) models. The optimized models were verified by introducing simulated constant errors (CE) and proportional errors (PE) respectively. The false positive alarm rate (FAR) was used to evaluate specificity, and the average number of patients before error detection (ANPed) was used to evaluate sensitivity. According to the daily test volume and quality target requirements, we comprehensively judged whether the performance evaluation indicators of FAR and ANPed meet the laboratory requirements. Bias detection curve was used for determination of the best model.Results:The parameters of the best PBRTQC model for valproic acid drug concentration monitoring are: trimmed mean method with 1.5 standard deviations (i.e., truncating data outside 1.5 standard deviations of the data mean), λ=0.01. The performance verification result shows that ANPed of CE and PE of this model are both less than 100. The comparison between the EQA results and the EWMA results show that the EWMA method results are comparable to the EQA results.Conclusion:A PBRTQC model for the valproic acid drug concentration monitoring project based on the EWMA method has been successfully established. It is comparable with both IQC and EQA results, which means PBRTQC may be used as a supplement to the quality control of daily quality control products.

Objective:To explore the effect of Annexin A1(ANXA1)peptidomimetic Ac2-26 on acute kidney injury(AKI)and neutrophil apoptosis in septic rats.Methods:Experimental groups included control group,Ac2-26 group,AKI group,AKI+Ac2-26 group,with 15 rats in each group.After cecal ligation and perforation were used to establish a sepsis-induced AKI model,Ac2-26 was intravenously infused for treatment,once a day for 14 days;after the end,ELISA was used to detect levels of serum creatinine(Scr),urea nitrogen(BUN),IL-1β,IL-6 and TNF-α;HE staining and periodic acid Schiff(PAS)staining were used to observe the pathological changes of rat kidney tissues in each group;immunohistochemical staining was used to detect expression of ANXA1 in kidney tissue of each group of rats;neutrophils were isolated from rat peripheral blood,Giemsa staining and trypan blue staining were used to detect cell purity and viability;Annexin V-FITC/PI double staining method and TUNEL staining were used to determine apop-tosis level of neutrophils in each group.Results:Compared with control group,levels of Scr and BUN in serum of rats in AKI group were increased(P<0.05),levels of IL-1β,IL-6 and TNF-α also increased(P<0.05),renal tubules and glomeruli in kidney tissue were both significantly damaged,accompanied by a large number of inflammatory cell infiltration,and pathological score increased(P<0.05),while proportion of ANXA1 positive staining area was decreased(P<0.05);neutrophils identified by Giemsa staining and trypan blue staining had complete morphology and high activity;compared with control group,apoptosis rate of neutrophils in AKI group was decreased(P<0.05),and the positive rate of TUNEL was decreased(P<0.05).Compared with AKI group,levels of Scr and BUN in serum of rats in AKI+Ac2-26 group were decreased(P<0.05),levels of IL-1β,IL-6 and TNF-α also decreased(P<0.05),pathological manifestations of renal tubules and glomeruli in renal tissue were significantly reduced,and pathological score was reduced(P<0.05),while the proportion of ANXA1 positive staining area was increased(P<0.05),at the same time,apoptosis rate of rat neu-trophils was increased(P<0.05),positive rate of TUNEL was also increased(P<0.05).Conclusion:ANXA1 peptidomimetic Ac2-26 can increase expression of ANXA1 in kidney tissue of AKI in septic rats,promote neutrophil apoptosis,and have a protective effect on kidney tissue damage in rats caused by sepsis.

Objective:To explore the antagonistic effect of Fengzhecao extract against human red blood cell (RBC) hemolysis induced by wasp venom.Methods:Water extract method was used to extract dried Fengzhecao and vacuum-dried to obtain Fengzhecao extract. It was diluted into 1 g/L for next use. Wasp venom was collected from the wasp workers. A, B, O, AB type healthy blood donors' suspended RBC solution was obtain to make washed RBC solutions and adjust the RBCs count (4.0-80.0)×10 9/L (the number of RBC counted on the hemocytometer is 1-20 cells/small checker). According to treatment factors, they were divided into the normal saline controlled group (NS group; 200 μL RBC solution+20 μL normal saline), Fengzhecao extract group (FZC group; 200 μL RBC solution+10 μL Fengzhecao extract+10 μL normal saline), wasp venom group (FD group; 200 μL RBC solution+10 μL wasp venom+10 μL normal saline), and Fengzhecao extract+wasp venom group (FCD group; 200 μL RBC solution+10 μL Fengzhecao extract+10 μL wasp venom), with 10 blood samples per group of every blood type. The solutions were put into the glass test tube respectively, and then into 37 ℃ water bath thermostat. After 10 minutes, the blood cell counting plate was directly observed under the microscope and the RBCs was counted. Differences in RBC count was compared between the same treatment factors of different blood types and between different treatment factor groups of the same blood type. Results:There was no statistically significant difference in RBC count between blood types under the same treatment factors. The RBC count (×10 9/L) of the type A, B, O, AB in the NS group were 5.567±1.368, 5.146±1.690, 4.577±0.774, 5.197±1.587 ( F = 0.852, P = 0.475), the FZC group were 5.751±1.489, 5.268±1.418, 4.727±1.174, 5.298±1.229 ( F = 0.987, P = 0.410), the FD group were 0.546±0.450, 0.804±0.428, 0.679±0.283, 0.846±0.453 ( F = 1.089, P = 0.366), and the FCD group were 5.532±1.330, 5.051±1.596, 4.589±0.879, 5.140±1.492 ( F = 0.820, P = 0.492), respectively. Comparison of RBC count between groups with different treatment factors of the same blood type was done. There was no significant difference between the FZC group and the NS group, indicating that the extract of Fengzhecao extract had no effect on hemolysis of RBC; in the FD group, it was significantly lower than the NS group (all P < 0.05), indicating that wasp venom had a significant hemolytic effect on RBC; but there was no statistically significant difference in RBC count between the FCD group and the NS group, indicating that the Fengzhecao extract antagonizes the hemolytic effect of wasp venom without affecting the RBC count; however, the RBC count in the FCD group was significantly higher than that in the FD group (all P < 0.05), further indicating that the Fengzhecao extract antagonizes the hemolytic effect of wasp venom. Conclusion:Wasp venom has a significant hemolytic effect which can be effectively antagonized by Fengzhecao extract and has nothing to do with the human ABO blood type.

Objective:To establish a candidate reference method for simultaneous determination of whole blood iron and copper based on ICP-MS technology.Methods:Cobalt (Co) was chosen as the internal standard, and was added gravimetrically into the standard solution and the whole blood sample. The whole blood sample was digested by electric heating plate and diluted. The isotopic ratios of 57Fe/ 59Co and 63Cu/ 59Co were measured by ICP-MS in the isotopic analysis mode. The precision, standard recovery and accuracy of the method were verified by measuring EQA materials and certified standard materials, and the performance of the method was further evaluated by method comparison. Results:The detection limit of iron is 0.136 mg/kg and the limit of quantification was 0.454 mg/kg in this method. The detection limit of copper in whole blood was 0.008 mg/kg and the limit of quantification was 0.027 mg/kg. The standard curve of copper ranges from 0.83-3.33 mg/kg, iron ranges from 167-667 mg/kg. The calibration curve of iron and copper was good linearity ( R 2>0.999 90). The precision of the method did well. The total CV range was 0.42%-0.68% for iron and 0.14%-0.94% for copper. The spike recoveries were all around 100%, the range of iron: 99.69%-100.07%; copper: 99.26%-100.51%. The correlation between this method and the existing clinical mass spectrometry methods was good. Conclusions:A candidate reference ICP-MS method for simultaneous determination of whole blood Fe and Cu was established. This is a simple and rapid method with good precision and spike recovery compared to the traditional one.

Objective To perform a methodological evaluation study of 4 HPLC based systems and a capillary electrophoresis based system , with the International Federation of Clinical Chemistry and Laboratory Medicine ( IFCC) glycated hemoglobin reference method as a comparative method .Methods 40 hemolysis samples of variety concentrations were prepared .The samples were measured by IFCC reference method and 5 glycated hemoglobin testing systems , respectively and trueness verification was performed according to the Clinical &Laboratory Standards Institute (CLSI) guideline EP9-A3.Whole blood samples were used to test the systems'precision, linearity and analytical interferences .Results The average CV of IFCC reference method results of 40 hemolysis samples was 1.4％( range from 0.2％to 2.5％).No outlier was found in the results of the 5 testing system.The slopes ranged from 0.9902 to 1.0267, and intercepts from -0.1526 mmol/mol to +0.1512 mmol/mol, squared correlation coefficient from 0.9962-0.9971, biases at two medical decision level were less than 0.3％HbA1c.Within-laboratory precisions were less than 2％ (NGSP unit).Bias between all test results and predicted results were less than 5％.High concentration of glucose showed certain interference to glycated Hemoglobin tests but had little influence in clinical practice.Conclusions The results of the 5 testing system are comparable to the IFCC reference method , the results of precision and linearity evaluation meet the requirements of related guidelines .