Intravenous immunoglobulin (IVIg) is widely used to treat autoimmune diseases, yet its precise mechanisms of action in systemic lupus erythematosus remain incompletely understood. Here, we employed an integrated multi-modal approach combining single-cell RNA sequencing, quantitative proteomics, flow cytometry, immunofluorescence, enzyme-linked immunosorbent assay, and histopathological analysis to investigate IVIg-induced immune reprogramming in MRL/lpr lupus-prone mice. IVIg treatment markedly alleviated disease manifestations, including splenomegaly, proteinuria, renal injury, and systemic autoantibody production. Histological examination of the spleen and kidney showed decreased lymphoid hyperplasia, glomerular inflammation, and tissue damage in IVIg-treated mice. At the immune-cell level, IVIg was associated with alterations in T and B cell activation states, partial restoration of immune repertoire diversity, and shifts in dendritic cell populations. Collectively, our integrated analyses suggest that IVIg treatment is correlated with broad immunomodulatory changes in splenic immune composition in lupus-prone mice.

Background:Clinical therapeutic human serum albumin (HSA) preparations are typically derived from human plasma and contain various accompanying proteins (APs). Previous studies have documented extensively the disparities in post-translation modifications, redox states and antioxidant capacities among HSA preparations from different manufacturers. Most of these studies have focused primarily on albumin, and analyzing APs in HSA preparations and recombinant HSA (rHSA) was often neglected. Methods:In this study, the APs in human plasma-derived HSA (pHSA) from six Chinese manufacturers and recombinant HSA (rHSA) from yeast and rice were identified and analyzed using a four-dimensional (4D) label-free quantitative proteomic technology. Results:A total of 456 different APs from the six pHSA preparations were identified, with 96 APs consistently detected in all pHSA samples. 52 APs from yeast-produced rHSA were identified, whereas 152 APs were detected in rice-expressed rHSA. Among the detected APs, haptoglobin, hemopexin and transthyretin were among the top eight APs with the highest relative abundance consistently observed in all pHSA preparations. Moreover, the results revealed that the identified APs in pHSA are primarily involved in endopeptidase inhibitor activity, complement and coagulation cascades, biosynthesis of amino acids and cholesterol metabolism by Gene Ontology (GO), Clusters of Orthologous Groups (COG)/euKaryotic Orthologous Groups (KOG), Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein-protein interactions (PPI) annotation. The ELISA validation results confirmed the presence of haptoglobin, hemopexin, transthyretin and serotransferrin in pHSA but not in rHSA, aligning with the findings from the 4D label-free quantitative proteomic analysis.

Colorectal cancer (CRC) is the third most prevalent malignancy worldwide. Inflammatory bowel diseases (IBD) encompass a group of chronic intestinal inflammatory disorders, including ulcerative colitis (UC) and Crohn's disease (CD). As a chronic inflammatory bowel disease, UC may persist and elevate the risk of malignancy, thereby contributing to the development of colorectal cancer, known as colitis-associated colorectal cancer (CAC). Chronic intestinal inflammation is a significant risk factor for colorectal cancer, and the incidence of colitis-associated colorectal cancer continues to rise. Current studies indicate that therapeutic agents targeting inflammation and key molecules or signaling pathways involved in the inflammatory process may effectively prevent and treat CAC. Mechanistically, drugs with anti-inflammatory or modulatory effects on inflammation-related pathways may exert preventive or therapeutic roles in CAC through multiple molecules or signaling pathways implicated in tumor development. Moreover, the development or discovery of novel drugs with anti-inflammatory properties to prevent or delay CAC progression is becoming an emerging field in fighting against CRC. Therefore, this review aims to summarize drugs that prevent or delay CAC through modulating anti-inflammatory pathways. First, we categorize the published studies exploring the role of anti-inflammatory in CAC prevention. Second, we highlight the specific molecular mechanisms underlying the anti-inflammatory effect of the above-mentioned drugs. Finally, we discuss the potential and challenges associated with clinical application of these drugs. It is hoped that this review offers new insights for further drug development and mechanism exploration.

Treating bacteremia caused by antibiotic-resistant bacteria is a global concern. Antibacterial photodynamic inactivation is a promising strategy to combat it. However, it’s challenging to achieve the inactivation of antibiotic-resistant bacteria in whole blood because of its opacity and complexity. We investigated a riboflavin photodynamic method to effectively inactivate antibiotic-resistant bacteria in whole blood. Four strains of antibiotic-resistant bacteria were isolated, identified, and cultured in this research: methicillin-resistant Staphylococcus aureus (MRSA), pan-drug-resistant Acinetobacter baumannii (PDRAB), ESBLs-producing Escherichia coli (EPEC) and pan-drug-resistant Klebsiella pneumoniae (PDRKP). To simulate bacteremia, antibiotic-resistant bacteria was added into whole blood. Whole blood was treated using riboflavin photodynamic method with ultraviolet irradiation (308 nm and 365 nm). The ultraviolet irradiation dose was divided into 18 J/cm2, 36 J/cm2, and 54 J/cm2. Microbial count of antibiotic-resistant bacteria in whole blood was used for evaluating inactivation effectiveness. The roles of red blood cells, lymphocytes, coagulation factors, and platelets in whole blood were assessed. In results, inactivation effectiveness increased as the ultraviolet dose increased from 18 J/cm2 to 54 J/cm2. At the dose of 18 J/cm2, inactivation effectiveness of four antibiotic-resistant bacteria were more than 80%, while only 67% of MRSA. The antibacterial effect was enhanced by the combination of riboflavin photodynamic treatment and antibiotic. The red blood cell function was susceptible to ultraviolet dose. At the dose of 18 J/cm2, hemolysis rate was less than 0.8% and there was no change in levels of ATP and 2,3-DPG. At the same dose, the proliferation, cell killing, and cytokine secretion activities of lymphocytes decreased 20–70%; Factor V and Factor VIII activities decreased 50%; Fibrinogen and platelet function loss significantly but reparable. Consequently, we speculated that riboflavin photodynamic method with a ultraviolet dose of 18 J/cm2 was effective in inactivating four antibiotic-resistant bacteria in whole blood while whole blood function was preserved. We also provided a novel extracorporeal circulation phototherapy mode for treating bacteremia caused by antibiotic-resistant bacteria.

Radiation exposure often leads to serious health problems in humans. The intestinal epithelium is sensitive to radiation damage, and radiation causes destruction of the intestinal epithelial barrier, which leads to radiation enteritis (RE), the loss of fluids, and the translocation of intestinal bacteria and toxins; radiation can even threaten survival. In this study, we aimed to explore the influence of IVIg on the integrity of the intestinal epithelial barrier after RE. Using a RE mouse model, we investigated the protective effects of intravenous immunoglobulin (IVIg) on the epithelial junctions of RE mice and validated these findings with intestinal organoids cultured in vitro. In addition, transmission electron microscopy (TEM), western blotting (WB) and immunostaining were used to further investigate changes in intestinal epithelial ferroptosis and related signaling pathways. When RE occurs, the intestinal epithelial barrier is severely damaged. IVIg treatment significantly ameliorated this damage to epithelial tight junctions both in vivo and in vitro. Notably, IVIg alleviated RE by inhibiting intestinal epithelial ferroptosis in RE mice. Mechanistically, IVIg promoted activation of the mTOR pathway and inhibited ferroptosis in the intestinal epithelium of mice. Rapamycin, which is a potent inhibitor of the mTOR protein, significantly abolished the protective effect of IVIg against radiation-induced damage to intestinal epithelial tight junctions. Overall, IVIg can prevent RE-induced damage to the intestinal epithelial barrier and inhibit ferroptosis by activating the mTOR pathway; this study provides a new treatment strategy for patients with RE caused by radiotherapy or accidental nuclear exposure.

Background As a group of more than 3.67 million people, the bone health of Chinese plasmapheresis donors, which the main population is also a risk group of osteoporosis (OP), has raised concerns. Therefore, this article investigates the relationship between bone mineral density (BMD), bone metabolism indicators, and plasmapheresis donation behavior among some high-risk plasmapheresis donors for OP in China, and further explores the mediating factors through reasonable statistical methods. Methods Recruiting long-term and highly active plasmapheresis donors and new donors to measure the total calcium, albumin (ALB), 25-hydroxy vitamin D (25OHD), parathyroid hormone (PTH), type I procollagen amino-terminal peptide (P1NP), and type I collagen carboxy-terminal peptide (β-CTX) and serum ferritin (SF). Then, multiple linear regression was used to adjust confounding factors. Using restrictive cubic splines to explore the nonlinear relationship. Using the Bootstrap method, investigate whether SF has a mediating effect between plasmapheresis donation behavior and bone metabolism biomakers. Finally, analyze the differences in BMD between the two. Results Compared to new donors, repeat donors have a lower 25OHD, β-CTX and SF levels, while P1NP and PTH levels are high, with statistical differences. The Bootstrap analysis results show that SF level is a partial mediating factor between plasmapheresis donation behavior and bone metabolism biomakers, with a mediating effect ratio of 21.8%. There was no significant difference in the BMD between the two. Conclusion Long-term and frequently plasmapheresis donation does not affect the bone mass of even elderly donors at high risk for osteoporosis under the existing collection standards and anticoagulant use in China. However, as a self-regulation way, it does increase the osteogenic activity of the body.

Abnormal plasma uric acid (UA) levels, the lipid profile, and plasma proteins in blood are associated with a range of adverse health outcomes. This multicenter, prospective cohort study aimed to determine the possible effects of multiple apheresis plasma donations on plasma UA levels, the lipid profile, and major proteins in plasma donors. Participants were enrolled from 1 April 2021 to 31 August 2022. When their plasma UA (men: >420 µmol/L, women: >360 µmol/L) and/or lipid levels (total cholesterol [TC]: ≥6.2mmol/L, triglycerides [TGs]: ≥2.3mmol/L, low-density lipoprotein cholesterol: ≥4.1mmol/L, or high-density lipoprotein cholesterol [HDL-C]: <1.0mmol/L) were abnormal at their first plasma donation, the enrolled participants were followed up until they had completed 10 plasma donations. A total of 11485 participants were enrolled, of whom 1861 met the inclusion criteria. During the study period, 320 donors completed 10 plasma donations. None of the participants took any corrective medicine for their abnormal index. The measured parameters were significantly different from the first to the tenth plasma donations (donors with asymptomatic hyperuricemia: UA, P<0.001; donors with asymptomatic hyperlipidemia: HDL-C, P<0.001; TC, P=0.025; TGs, P<0.001; apolipoprotein B, P=0.025; all of the plasma donors, immunoglobulin G, P<0.001). The levels of HDL-C, TC, and apolipoprotein B were increased, and the levels of UA, TGs, and immunoglobulin G were decreased over this time. However, immunoglobulin G levels were still in the normal range. Moreover, the changes in these parameters were closely associated with the frequency of plasma donation during the study period. Repeated apheresis plasma donations can reduce plasma UA and TG levels and increase HDL-C levels; and further evaluation of the clinical significance with a larger sample size is required.

Background Endovascular thrombectomy has been confirmed to be an effective therapy for acute ischemic stroke (AIS). However, how functional brain networks reorganize after restoration of blood supply in AIS patients, and whether the degree of reperfusion associates with functional network changes remains unclear. Methods Resting-state fMRI data were collected from 43 AIS patients with anterior circulation occlusion after thrombectomy and 37 healthy controls (HCs). Both static and dynamic functional connectivity (FC) within four advanced functional networks including dorsal attention network (DAN), ventral attention network (VAN), executive control network (ECN) and default mode network (DMN), were calculated and compared between post-thrombectomy patients and HCs, and between two subgroups of post-thrombectomy patients with different reperfusion conditions. Results As compared to HCs, patients showed significant differences in static FC of four functional networks, and in dynamic FC of DAN, ECN and DMN. Furthermore, patients with better reperfusion conditions exhibited increased static FC with precuneus, and altered dynamic FC within precuneus. Moreover, these alterations were associated with clinical assessments of stroke severity and functional recovery in post-thrombectomy patients. Conclusions Collectively, these findings may provide the potential imaging markers for assessment of thrombectomy efficacy and help establish the specific rehabilitation treatments for post-thrombectomy patients.

Background: Mechanism of action of intravenous immunoglobulin (IVIG) is complex, and involves the immunoglobulin G (IgG) Fc fragment binding to Fc receptors on immune cells as activators and inhibitors. Antibody -dependent cell -mediated cytotoxicity (ADCC) is a mechanism that mainly depends on Fc gamma RIIIA. However, the IVIG Fc fragment -binding activity to Fc gamma RIIIA is unknown, and the IVIG-mediated ADCC remains to be clarified. This study attempts to explore the IVIG-mediated ADCC and provide correlative evidence. Methods: This study evaluated the IVIG-mediated ADCC using a time -resolved fluorescence (TRF) cell cytotoxicity assay in vitro , and Jurkat-NFAT-Luc-CD16 cells were used to detect the IVIG Fc fragmentbinding activity to Fc gamma RIIIA. Quantitative real-time polymerase chain reaction (PCR) analyses were conducted to detect mRNA synthesis when IVIG incubated with effector cells, and the final expression of Fc gamma RIIIA on the cell membranes were detected by flow cytometry. Results: The results showed that IVIG had optimal binding activity to Fc gamma RIIIA fitted with a fourparameter curve equation regression model with R 2 =0.99 and caused specific ADCC of about 40%. Meanwhile, IVIG upregulated the expression of FCGR3A by two -fold, and the Fc gamma RIIIA level was markedly increased from 33.7% up to 66.7%. Conclusions: This study investigated the IVIG-mediated ADCC, including measurement of the IVIG Fc fragment -binding activity to Fc gamma RIIIA and the regulation of Fc gamma RIIIA expression by IVIG. The results indicate that ADCC that can kill tumor cells and IVIG can also promote FCGR3A synthesis, increasing Fc gamma RIIIA expression on immune cells membranes. The findings provide further evidence for the IVIGmediated ADCC and understanding of the biological activity of IVIG.

Chemotherapy-induced mucositis (CIM) significantly impacts quality of life and reduces survival in patients treated with specific chemotherapeutic agents. However, effective clinical treatments for CIM remain limited. Intravenous immunoglobulin (IVIg), a therapeutic derived from pooled human plasma, is widely used to treat inflammatory diseases. This study aimed to evaluate the therapeutic efficacy and underlying mechanisms of IVIg in CIM. A murine model of doxorubicin (Dox)-induced intestinal mucositis and an organoid model of small intestinal injury were used to explore the protective effects of IVIg on CIM. Immunostaining, transmission electron microscopy (TEM), western blotting (WB), and proteomic analysis were used to further investigate ferroptosis in intestinal epithelial cells and the underlying mechanisms. In the murine model of Dox-induced intestinal mucositis, intestinal epithelial barrier was destroyed and ferroptosis increased, characterized by weight loss, hematological injury, inflammation, mitochondrial atrophy in intestinal epithelial cells, lipid peroxidation, impairment of tight junctions, and damage to intestinal microvilli. IVIg treatment significantly ameliorated intestinal epithelial barrier damage and reduced ferroptosis both in vitro and in vivo. Proteomic analysis revealed that the FcγR-mediated phagocytosis signaling pathway was involved in the therapeutic effects of IVIg on CIM mice. WB results demonstrated that key proteins downstream of this pathway, Syk, PI3K, and Akt, showed increased phosphorylation in CIM mice, whereas IVIg treatment significantly reduced the phosphorylation levels. Furthermore, the inhibitory effects of IVIg on Dox-induced activation of the Syk/PI3K/Akt axis and ferroptosis, as well as its protective effects on intestinal inflammation and intestinal barrier damage, were reversed by 740Y-P (an PI3K activator) or SC79 (an Akt activator). Our findings highlight that IVIg ameliorates CIM by inhibiting ferroptosis via the Syk/PI3K/Akt axis. These results suggest that IVIg may represent a potential therapeutic approach for CIM.

Background China’s plasmapheresis donation policy differs from that of Western countries. The association between regular plasmapheresis donation and donor health in China is still unknown.Objectives To investigate the association of regular plasmapheresis donation with serum protein and electrolyte levels and provide scientific evidence for policy improvement.Design Multicentre cross-sectional study.Setting and participants A total of 767 regular and 726 new donors from the provinces of Sichuan, Hunan, Henan and Yunnan were recruited from September 2021 to October 2022.Primary and secondary outcome measures Our primary outcome focused on measuring the levels of serum protein and electrolyte levels, including total serum protein (TSP), IgG, albumin (Alb), haemoglobin (Hb), calcium, potassium (K+) and magnesium (Mg2+). The secondary outcome assessed their abnormal rates.Results Male and female donors in the high donation frequency group (>16 donations per year) exhibited lower IgG levels compared with new donors (p=0.008 for male donors and p=0.007 for female donors). Additionally, female donors with high donation frequency and a high total number of lifetime donations (>100 donations) had significantly lower Hb concentrations than new donors. However, no significant changes were observed in TSP, Alb, calcium, K+ and Mg2+ levels. There were also no statistically significant differences in the rates of abnormal protein and electrolyte values below the respective threshold levels between new and regular donors.Conclusions Plasmapheresis donation is not associated with an increased risk of abnormalities in the analysed parameters. However, the results provide preliminary evidence supporting the routine inclusion of IgG screening for donors, as plasmapheresis donation is associated with a decrease in IgG levels. Particular attention should be paid to the Hb levels of female donors, especially those who donate frequently. Testing of TSP at each donation may not be necessary.

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.

Objective To investigate the level of serum bone metabolism and biochemical markers and bone densi-ty of plasmapheresis donors,and to provide scientific basis for ensuring the health and safety of plasmapheresis donors in China.Methods A total of 437 plasmapheresis donors from Linwu plasmapheresis station in Hunan Province from July 1 to September 30,2022 were recruited to determine the levels of total serum calcium,albumin,serum 25-hy-droxyvitamin D(25OHD),serum type Ⅰ procollagen N-terminal propeptide(P1NP),and collagen type 1 crosslinked carboxy l-terminal peptide(β-CTX).Dual-energy X-ray method was used to measure the bone density of the anteroposte-rior lumbar spine(L1-L4)and bilateral femoral neck bone density of plasmapheresis donors.Plasmapheresis donors were grouped according to the type of plasma donation(first-time and repeat plasmapheresis donors)and the total num-ber of plasma donations to assess the differences in bone density and serum bone metabolism biochemical markers be-tween groups.The dose-response relationship between the total number of plasmapheresis donations and biochemical in-dexes was analyzed by limiting cubic spline,and the influencing factors of different indexes were explored by multiple linear regression.Results A total of 437 plasmapheresis donors were included in this study,including 187 first-time plasmapheresis donors and 250 repeat plasmapheresis donors.There were no significant differences in bone density and prevalence of osteoporosis between first-time donors and repeat donors(P＞0.05).There was also no significant differ-ence in bone density levels between groups of total number of plasmapheresis donations.The levels of albumin and 25OHD decreased with the increase of the total number of plasma donations,while the serum P1NP level was positively correlated with the total number of plasma donations.The results of the restriction cubic spline showed that the total num-ber of plasmapheresis donations had a nonlinear dose-response relationship with 25OH and P1NP(P＜0.05).The results of multiple linear regression showed that the frequency of plasmapheresis donation was the influencing factor of 25OHD,and the total number of plasmapheresis donation was the influencing factor of P1NP.Conclusion Plasmapher-esis donation does not affect the bone health of donors and increase the risk of osteoporosis due to the use of long-term anticoagulants,but it will increase the osteogenic activity of plasmapheresis donors.It is recommended that middle-aged and elderly plasmapheresis donors supplement vitamin D appropriately.

Objective The purpose of the present study was to evaluate whether intravenous immunoglobulin (IVIG) increases live birth rates and improves neonatal results in patients with antiphospholipid antibodies (aPL) at high-risk for miscarriage. Background Positivity of aPL in pregnant women is a high-risk factor for miscarriage, and IVIG treatment has emerged as a potential intervention. Methods The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline was employed to search multiple electronic databases for articles published until August 20, 2023, including PubMed, Web of Science, Embase, Scopus and Medline. The inclusion criteria encompassed studies assessing the efficacy of IVIG in aPL-positive patients with a high risk of miscarriage. Relevant articles were assessed for the quality and data were extracted for analysis. Two independent reviewers performed study selection, data extraction, and quality assessments. The risk of bias was evaluated according to the Cochrane risk of bias tool. All analyses were conducted using Review Manager 5.3. Results This systematic review included nine randomized controlled trials, with 366 aPL-positive women at high risk of miscarriage. These studies included in this review were randomized controlled trials. The primary outcome measures were successful pregnancy outcomes and live birth rates. The secondary outcomes included obstetric complications, and neonatal outcomes (such as birth weight and live-birth rate). The comparison between the intervention and control groups revealed no significant differences in terms of obstetric complications and neonatal outcomes. The group receiving IVIG treatment had a higher prevalence of preterm deliveries than controls (OR = 2.05, I2 = 46%, 95% CI [0.58–5.24]), but also exhibited a partial improvement in live birth rates (OR = 2.86, I2 = 52%, 95% CI [1.04–7.90]), because it reduced the number of miscarriages (OR = 0.35, I2 = 52%, 95% CI [0.13–0.96]). Conclusion Based on the available evidence, IVIG intervention appears to be a potentially effective approach for managing of aPL-positive pregnant women with high risk of miscarriage. While IVIG shows significant potential in tripling the chances of having a live-born child, further large-scale randomized controlled trials are necessary, preferably comparing IVIG with hydroxychloroquine or lifestyle and dietary interventions, to refine treatment protocols and ensure the most effective application.

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,

BACKGROUND:The variations in sequence, three-dimensional structure, and post-translational modifications (PTMs) of human serum albumin (HSA) are crucial for its physiological functions. This study aims to analyze and compare the disparities in PTMs between HSA derived from human plasma and genetically recombinant sources for clinical treatments in China. METHODS:Six distinct PTMs, namely acetylation, succinylation, crotonylation, phosphorylation, beta-hydroxybutyrylation, and lactylation, were identified using pan-specific antibodies via Western blot analysis. The samples, comprising human plasma-derived HSA (pHSA) from six different manufacturers and recombinant HSA (rHSA) expressed in yeast and Oryza sativa, underwent detection for various types of PTMs. Additionally, a 4D label-free quantitative proteomic analysis was performed to identify N-glycosylation and the aforementioned PTMs in both pHSA and rHSA samples. This analysis aimed to discern disparities in modification sites and levels. RESULTS:Through Western blot analysis, all six pHSA and two rHSA samples displayed positive bands for albumin (66.5 kDa) across the six PTMs. Subsequent analysis using 4D label-free quantitative proteomics revealed 25 (29) acetylated, 30 (32) succinylated, 41 (50) malonylated, 15 (23) phosphorylated, 36 (30) beta-hydroxybutyrylated, and 27 (34) lactylated modification sites in pHSA and rHSA samples, with no N-glycosylation modification sites detected. The analysis identified 1 acetylation (ALB_K160), 2 beta-hydroxybutyrylation (ALB_K569, ALB_K426), and 3 crotonylation (ALB_K264, ALB_K581, ALB_K560) specific modification sites in pHSA, as well as 3 crotonylation (ALB_K560, ALB_K562, ALB_K75), 1 succinylation (ALB_K490), and 23 phosphorylation specific modification sites in rHSA. In pHSA (rHSA), 2 (6) acetylation, 10 (12) succinylation, 0 (9) crotonylation, 1 (9) phosphorylation, 6 (0) beta-hydroxybutyrylation, and 0 (7) lactylation specific modification sites were found. Moreover, in the shared modification sites between pHSA and rHSA, pHSA exhibited up-regulation of amberylation (16:1) and beta-hydroxybutyrylation (12:2) in more sites, and up-regulation of acetylation (7:11), crotonylation (2:11), phosphorylation (1:8), and lactylation (1:14) in fewer sites compared to rHSA. CONCLUSION:In clinical practice, both pHSA and rHSA utilized in China commonly display acetylation, succinylation, crotonylation, phosphorylation, beta-hydroxybutyrylation, and lactylation. Notably, there exist distinctions in the site characteristics and modification levels of these alterations between pHSA and rHSA. Further experimental inquiries are imperative to delve into the implications of these disparities in PTMs on the biological functionality, effectiveness, and safety of pHSA and rHSA.

BACKGROUND AND OBJECTIVES:The present study aims to evaluate the iron stores in plasmapheresis donors and develop and validate an iron deficiency (ID) risk prediction model for plasmapheresis donors with potential or existing ID. MATERIALS AND METHODS:We assessed plasmapheresis donors' serum ferritin (SF) and haemoglobin (Hb) levels. The candidate factors showing significant differences in the multivariate logistic regression analysis were used to establish a risk prediction scoring system. The participants were divided into a training cohort and an internal validation cohort in a 7:3 ratio. Additional plasmapheresis donors from a different station were recruited for external validation. RESULTS:The SF levels in both male and female donors in the high-frequency group were significantly lower than those of new donors (male: p < 0.001; female: p = 0.008). The prevalence of ID in female regular donors with a high frequency was significantly higher than that in new donors (33.1% vs. 24.6%; odds ratio = 1.209 [95% CI: 1.035-1.412]). Donation frequency, age, Hb, body mass index and being pre-menopausal were identified as independent risk factors for ID (p < 0.05). The developed model exhibited good discrimination ability (area under the receiver operating characteristic curve >0.7) and calibration (p > 0.05) in development, internal validation cohorts and external validation cohorts. CONCLUSION:A higher donation frequency has been associated with reduced SF levels and an increased risk of ID in women. The developed ID risk prediction model demonstrates moderate discriminative power and good model fitting, suggesting its potential clinical utility.

目的 了解单采血浆对献浆者部分血浆蛋白和电解质水平的影响,为保障献浆者健康安全提供有力的科学依据.方法 2022年2月—5月在四川省两家单采血浆站招募223名新献浆者,经纳排后共纳入118名献血浆者.收集献浆者人口统计学信息,并持续随访观察一年,检测每次捐献血浆前部分血浆蛋白及电解质水平.分析部分血浆蛋白及电解质水平的变化趋势,利用协方差分析排除献浆前各相关指标基线水平的干扰,分析献浆次数对献浆后相关指标变化的影响,并采用多重线性回归探索相关指标的影响因素.结果 男性献浆者铁蛋白(SF,serum ferritin)水平随献浆次数增加呈现一定下降趋势,其余成分水平则随献浆次数增加无明显波动.协方差分析控制混杂因素后,献浆次数对SF的影响无统计学意义.多重线性回归结果显示,在控制了基线水平之后,献浆次数对SF水平的影响无统计学意义.结论 持续一年低频次捐献单采血浆不会对献浆者血浆蛋白及电解质水平产生统计学上的显著影响.

Neuroinflammation is closely related to prognosis in ischemic stroke. Microglia are the main immune cells in the nervous system. Under physiological conditions, microglia participate in clearance of dead cells, synapse pruning and regulation of neuronal circuits to maintain the overall health of the nervous system. Once ischemic stroke occurs, microglia function in the occurrence and progression of neuroinflammation. Therefore, the regulation of microglia-mediated neuroinflammation is a potential therapeutic strategy for ischemic stroke. The anti-inflammatory activity of gypenosides (GPs) has been confirmed to be related to the activity of microglia in other neurological diseases. However, the role of GPs in neuroinflammation after ischemic stroke has not been studied. In this study, we investigated whether GPs could reduce neuroinflammation by regulating microglia and the underlying mechanism through qRT-PCR and western blot. Results showed that GPs pretreatment mitigated blood–brain barrier (BBB) damage in the mice subjected to middle cerebral artery occlusion (MCAO) and improved motor function. According to the results of immunofluorescence staining, GPs pretreatment alleviated neuroinflammation in MCAO mice by reducing the number of microglia and promoting their phenotypic transformation from M1 to M2. Furthermore, GPs pretreatment reduced the number of astrocytes in the penumbra and inhibited their polarization into the A1 type. We applied oxygen and glucose deprivation (OGD) on BV2 cells to mimic ischemic conditions in vitro and found similar effect as that in vivo . At the molecular level, the STAT-3/HIF1-α and TLR-4/NF-κB/HIF1-α pathways were involved in the anti-inflammatory effects of GPs in vitro and in vivo . Overall, this research indicates that GPs are potential therapeutic agents for ischemic stroke and has important reference significance to further explore the possibility of GPs application in ischemic stroke.

Objective To analyze the dynamic relationship between the setting up of plasmapheresis station and the volume of voluntary blood donation collected using panel vector autoregressive model, so as to provide scientific reference for the management policies of blood stations and plasmapheresis stations in China. Methods The data collected from blood stations in seven administrative regions of Guangyuan, Sichuan Province from 2011 to 2021, as well as plasma collection data from two plasmapheresis stations in the region within two years since their operation, were collected. A panel vector autoregressive model was constructed. Impulse response analysis and variance decomposition analysis were used to analyze the impact and time lag effects of simulated plasmapheresis station settings on the collection volume of voluntary blood donation. Covariance analysis was used to explore whether the establishment of plasmapheresis station had an impact on the volume of voluntary blood donation collected after excluding the impact of initial value differences. Results The pulse response results showed that after the plasmapheresis station was set up, there was a negative impact effect on the voluntary blood donation collection volume at the first stage, and its impact began to rise after the second stage, reached the highest value in the third stage, and then began to decline. After the seventh stage, it tended to be stable. However, within the 10 stage range, the confidence interval for the response strength of voluntary blood donation collection volume always included 0, indicating that the response of blood collection volume to the plasmapheresis station setting in the region was not statistically significant. The results of variance decomposition showed that the contribution of collection volume of voluntary blood donation to their own impact reached 94.3%. In terms of the contribution of plasmapheresis station factors, the number of plasma donors has a relatively greater impact on the volume of voluntary blood donation collected(2.2%). Covariance analysis showed that after removing the initial confounding factors, whether to establish a plasmapheresis station had no significant impact on blood donation volume in the two groups of regions (P>0.05). Conclusion The establishment of a new plasmapheresis station will have a certain impact on blood collection volume of blood stations in the region in a short term, but in the long term, it may not directly affect the voluntary blood donation collection in the region.