1.Desalination effect on FⅧ components: a compartive study among 5 desalination methods
Renjun PEI ; Xi DU ; Pan SUN ; Xiaoyue LI ; Peng JIANG ; Changqing LI ; Fangzhao LIN ; Haijun CAO
Chinese Journal of Blood Transfusion 2024;37(3):304-311
【Objective】 To compare the desalination effects of five desalination methods and their effects on the components for human coagulation factor Ⅷ(FⅧ), and provide reference for selection of protein desalination methods. 【Methods】 Sephadex G-25 Medium gel, Fractogel EMD BioSEC gel, ultrafiltration, room temperature dialysis and 4℃ dialysis were used to desalt human FⅧ. The desalination effect was evaluated by the removal rate of Na +, citrate ion and glycine. FⅧ protein recovery, FⅧ activity (FⅧ∶C), VWF antigen (VWF∶Ag), VWF activity(VWF∶Ac), VWF polymers and SDS-PAGE analysis before and after desalination were compared to evaluate the effect of desalination on FⅧ components. 【Results】 In terms of desalination effect, the removal rate of Na+ was the lowest in ultrafiltration desalination, while that of Fractogel EMD BioSEC gel was the highest [(97.90±0.06) % vs (99.82±0.07) %]. Except that there was no statistical significance between Sephadex G-25 Medium gel desalination and Fractogel EMD BioSEC gel desalination (P=0.90), the removal rates of the other four methods were statistically significant. The removal rate of glycine was the lowest in ultrafiltration desalination, wihle that of Fractogel EMD BioSEC gel desalination was the highest [(95.78±0.42) % vs (99.81±0.08) %]. Significant difference in glycine removal was noticed in ultrafiltration desalination, but not among the other four desalination methods. There was no significant difference in the removal rate of citrate ions among the five methods (P=0.85). For the effect of FⅧ components, FⅧ∶C, VWF∶Ag, VWF∶Ac and protein recovery rates of ultrafiltration desalination were the highest, with (18.34±1.99) IU/mL, (11.81±0.33) IU/mL, (12.26±0.58) IU/mL and (97.13±1.37) %, respectively. There was no significant change in VWF∶Ac/VWF∶Ag before and after desalination by the five methods. SDS-PAGE and VWF polymer analysis showed that different desalination methods had no significant impact on protein composition. 【Conclusion】 Although different desalination methods had no significant effect on the composition of FⅧ protein, the desalination effect was different. Moreover, different desalination methods had significant effects on protein recovery, FⅧ∶C, VWF∶Ag and VWF∶Ac. The selection of desalination methods should be more considered during protein processing,
2.Binding and carrying role of human serum albumin from various sources to sphingosine-1-phosphate
Qing LIU ; Yafei ZHAO ; Jun XU ; Lu CHENG ; Yuwei HUANG ; Xi DU ; Changqing LI ; Zongkui WANG ; Li MA
Chinese Journal of Blood Transfusion 2024;37(5):524-533
Objective To investigate the binding and carrying effects of human serum albumin(HSA)from various sources on sphingosine-1-phosphate(S1P).Methods Utilizing human plasma-derived HSA(pHSA)and recombinant HSA(rHSA)samples as the focal points of our investigation,LC-MS/MS technology was employed to meticulously compare and an-alyze the disparities in S1P content among the aforementioned samples.Subsequently,under physiological concentration condi-tions,S1P was directly introduced to HSA samples for loading processing,facilitating a comprehensive comparison of the bind-ing efficacy of HSA from different sources to S1P.Within a serum-free culture setting,HSA samples from various sources were co-cultured with HUVEC cells.The alterations in S1P content within the cell culture supernatant across different treatment groups were meticulously analyzed,allowing for a nuanced comparison of the S1P carry effects exerted by HSA from different sources on cells.The interaction between HSA and S1P molecules from different sources was analyzed and their affinity was cal-culated using surface plasmon resonance(SPR)technology.Furthermore,leveraging AutoDock Vina software and the Mol-prophet platform,the molecular docking analysis of HSA and S1P was conducted,aiming to predict the key binding pocket do-main of S1P within HSA.Results All pHSA samples exhibited detectable levels of S1P(ranging from 3.31±0.03 to 30.35±0.07 μg/L),with significant variations observed among pHSA samples from different manufacturers(P<0.001).Conversely,S1P was undetectable in all rHSA samples.Upon load treatment,the binding affinity of HSA from diverse sources to S1P dem-onstrated significant discrepancies(P<0.001),with rHSA exhibiting approximately double the average S1P loading compared to pHSA(ΔCrHSA=801.75±142.45 μg/L vs ΔCpHSA=461.94±85.73 μg/L;P<0.001,t=5.006).Co-culture treatment out-comes revealed a significant elevation in S1P concentration within the supernatant after 6 hours of co-culture across all HSA sample processing groups with HUVEC cells,while no changes were observed in the supernatant of the blank control group.Notably,significant differences in supernatant S1P concentration were observed among treatment groups at 6 h,12 h,and 24 h(P<0.001).SPR analysis unveiled a stronger affinity of pHSA for S1P compared to rHSA(KDpHSA-S1P:2.38E-06,KDrHSA-S1P:3.72E-06).Molecular docking analysis and binding pocket prediction suggested that the key binding pocket of HSA and S1P may reside in the IB subdomain of the HSA molecule.Conclusion HSA from various sources exhibits distinct binding and carrying effects on S1P,which appear to be closely associated with the IB subdomain of the HSA molecule.
3.Diversity of IgG autoantibodies of 4 Chinese intravenous immunoglobulins
Xiaochen YAN ; Li MA ; Zongkui WANG ; Changqing LI ; Tingting LEI ; Xi DU ; Shengliang YE
Chinese Journal of Blood Transfusion 2023;36(12):1102-1106
【Objective】 To develop methods to display the IgG autoantibody repertoire of intravenous immunoglobulin (IVIG) products, analyze the different types of antibodies and study the diversity of IgG autoantibody in 4 IVIG preparations from different Chinese manufacturers. 【Methods】 Two-dimensional gel electrophoresis and immunoblotting with human umbilical vein endothelial cell (HUVEC) proteins were used to demonstrate the IgG autoantibody repertoire and the human protein microarray with bioinformatics analysis was employed to profile the immune reactive autoantigens of the 4 IVIG preparations. 【Results】 The methods to showcase the autoantibody repertoire and study the antibody diversity of IVIG were successfully established. High-quality repertoires of IVIG autoantibodies and biological information about self-proteins that can be recognized were obtained. There was a significant difference in the recognition of the quantity and variety of the self-antigens by different IVIG products. The number of antibodies against HUVEC proteins in four products ranged from 241-386. The number of proteins recognized on the human protein chip ranged from 292-435, with 172 human self-proteins recognized by all four products. 【Conclusion】 Demonstration of antibody repertoire and protein chip technology can be used to analyze IVIG products′ IgG autoantibody repertoire. All four preparations tested in this study exhibited a broad spectrum of antibodies against HUVEC proteins and human proteome microarray, each product had its unique antibody repertoire characteristics.
4.The risk of Alzheimer′s disease transmitted by blood transfusion based on APP/PS1 double transgenic mice
Zhongsheng CHEN ; Zhangcheng FEI ; Renjun PEI ; Congchao QIAO ; Zongkui WANG ; Shengliang YE ; Li MA ; Rong ZHANG ; Changqing LI ; Xi DU ; Haijun CAO
Chinese Journal of Blood Transfusion 2023;36(5):396-403
【Objective】 To explore the risk of Alzheimer′s disease (AD) transmitted by blood transfusion. 【Methods】 There were 10 APP/PS1 mice of 3, 6 and 9 months old, half female and half male, and the cognitive and behavioral abilities of C57 mice of the same age were measured, and the blood of the oldest APP/PS1 mice with no behavioral changes were collected to detect the contents of Aβ
5.Effect of IgG dimer concentration on the binding ability of IgG Fc fragment in IVIG to THP-1 cell surface receptors
Tingting LEI ; Wei ZHANG ; Changqing LI ; Shengliang YE ; Xi DU ; Li MA
Chinese Journal of Blood Transfusion 2023;36(2):125-129
【Objective】 To investigate the effect of immunoglobulin G (IgG) dimer concentration of intravenous immunoglobulin (IVIG) on the binding ability of IgG Fc fragment to THP-1 cell surface receptors. 【Methods】 Firstly, protein purification and high performance liquid chromatography (HPLC) were used to prepare different concentrations of IgG dimers. After that, IgG dimer was added to IVIG to prepare IVIG containing different concentrations of IgG dimer. Finally, based on the method established in our laboratory, we analyzed the effect of IgG dimer concentration in IVIG on the binding ability of IgG Fc fragment to THP-1 cell surface receptors. 【Results】 When the concentration of IgG dimer in IVIG was 1.11%-10.30%, its binding ability to Fc receptors on the surface of THP-1 cell was 97.67%-135.33%, and this binding ability was positively correlated with the concentration of IgG dimer. When the IgG dimer concentration exceeded 13.22%, the binding ability had no correlation with the IgG dimer concentration. 【Conclusion】 A certain concentration of IgG dimer can promote the binding ability of the IgG Fc fragment in IVIG to receptors on the surface of THP-1 cells, which needs further verification from animal experiments and clinical data.
6.A child in radiation examination without protective equipment: A case analysis
Defeng DONG ; Jingjing GE ; Liwei DU
Chinese Journal of Radiological Health 2023;32(4):466-469
This paper reviews the investigation of examining a child without protective equipment exposed to an exceptionally large radiation field, in order to provide a reference for the investigation and handling of similar cases in the future. Based on the analysis of the way of obtaining evidence and the application of law, the authors put forward some suggestions, such as improving the standards, laws, and regulations related to radiation, enhancing the protective facilities, standardizing the law enforcement procedures, and strengthening the publicity and training of radiation hygiene. The health-related rights and interests of the examinees shall be effectively protected.
7.Preliminary study on the effect of IVIG with different Aβ antibody on cognitive function of Alzheimer′s disease model mice
Zhangcheng FEI ; Zhongsheng CHEN ; Renjun PEI ; Xi DU ; Shengliang YE ; Congchao QIAO ; Haijun CAO ; Changqing LI
Chinese Journal of Blood Transfusion 2022;35(7):683-689
【Objective】 To investigate the effect of intravenous immunoglobulin(IVIG) with different Aβ antibody content on the cognitive function of Alzheimer′s disease model mice. 【Methods】 IVIG from 8 domestic blood products companies were selected. Enzyme-linked immunosorbent assay(ELISA) was used to detect the content of Aβ
8.A case report of HLRCC syndrome combined Ⅳ inferior vena cava tumor thrombus
Kehao LI ; Yuancheng DU ; Xueping ZHANG ; Changqing LIU ; Li GE ; Yongxiang LI ; Liang QIAO
Chinese Journal of Urology 2021;42(10):782-783
Hereditary leiomyomatosis and renal cell cancer (HLRCC) syndrome is a rare hereditary disease and characterized by cutaneous leiomyoma, uterine leiomyoma and/or renal cell carcinoma, but rarely associated with vena cava embolism. We treated 1 case of HLRCC syndrome patients with inferior vena cava tumor emboli (Mayo grade Ⅳ), confirmed after genetic testing, the patient and her family refused further treatment. The patient died after two months of follow-up after discharge.
9.Efficacy analysis of von Willebrand factor in human coagulation factor FⅧ concentrates in China
Xi DU ; Li MA ; Zongkui WANG ; Shengliang YE ; Fengjuan LIU ; Rong ZHANG ; Na SU ; Fangzhao LIN ; Haijun CAO ; Changqing LI
Chinese Journal of Blood Transfusion 2021;34(6):574-578
【Objective】 To obtain the quality information of von Willebrand factor (vWF) in coagulation factor Ⅷ (FⅧ) concentrates in China. 【Methods】 FⅧ concentrates produced by 7 domestic blood product manufactures and 1 foreign manufacture were collected, then FⅧ and vWF contained in FⅧ concentrates were evaluated. 【Results】 The activity loss of vWF was more than 25% in 2 of the 7 domestic FⅧ concentrates. The ratio of vWF activity to FⅧ activity in FⅧ concentrates from different domestic manufactures was significantly different (P<0.05). The ratio in FⅧ concentrates prepared by C, D, F manufacturer was greater than 1, which was similar to that in willate@ approved abroad for the treatment of vWD. The ratio in FⅧ concentrates prepared by E manufacturer was greater than 0.7 and less than 1, and by A, B, G manufacturers was less than 0.5. In addition, the specific activities of FⅧ and vWF were significantly different among different FⅧ concentrates in China (P<0.05), and the specific activities of FⅧ and vWF were much lower than that of willate@. 【Conclusion】 The variation of vWF quality between domestic FⅧ concentrates and willate@ is mainly due to the different in vWF content. After the comprehensive consideration of various indicators, the FⅧ concentrates made by C and D manufacturers may be used in the treatment of vWD.
10.ASCVD indexes in Bama and Shimen area plasma donors, China
Rong ZHANG ; Zongkui WANG ; Wei ZHANG ; Fengjuan LIU ; Shengliang YE ; Xi DU ; Haijun CAO ; Li MA ; Changqing LI
Chinese Journal of Blood Transfusion 2021;34(6):634-638
【Objective】 To investigate the level of atherosclerotic cardiovascular disease (ASCVD) related indexes in plasma donors from longevity area, and explore its influencing factors. 【Methods】 1 027 plasma donors from longevity hotspot (Bama, Guangxi province) and 1 816 donors from non-longevity region (Shimen, Hunan province) who donated plasma during June to November 2018 were randomly selected. Triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDLC), low density lipoprotein cholesterol (LDLC), apolipoprotein A1 (Apo-A1), apolipoprotein B (Apo-B), and fructosamine (FUN) of the two groups were measured and statistically analyzed. 【Results】 Compared with the non-longevity region group, the TG, TC and FUN levels of longevity hotspot group were lower (1.41±0.96 vs 2.31±1.28, 3.89±0.92 vs 4.04±0.82, 176.65±26.60 vs 200.33±34.19; all P<0.05), but HDLC, LDLC, Apo-A1 and Apo-B levels were higher (1.11±0.32 vs 0.96±0.25, 2.53±0.70 vs 2.29±0.56, 1.56±0.28 vs 1.23±0.18, 0.80±0.27 vs 0.72±0.19; all P<0.05). The yield (%) of high TG(12.0 vs 40.01) and FUN(0.58 vs 2.48), low HDLC(24.63 vs 43.90) and Apo-A1(1.66 vs 22.56) were lower in longevity area than those in non-longevity region (all P<0.05), but high LDLC(2.73 vs 0.28) and Apo-B(4.09 vs 0.22) yield(%) were higher in longevity area group ( P<0.05). The levels of TC, HDLC, LDLC, Apo-A1 and Apo-B were significantly different by ages (all P < 0.01), presenting positively correlated with age, significantly by gender and nationality, and slightly by blood type. 【Conclusion】 The ASCVD indexes of plasma donors from Bama were different from those from Shimen. Age, nationality, gender and blood type of donors from Bama all had a certain influence on these indexes levels.

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