Objective: To establish evidence-based, safe and efficient management of HIV-reactive blood donors by investigating safe and feasible assessment strategies for HIV-reactive blood donors based on routine blood screening data. Methods: The data of blood screening, supplementary testing, follow-up and CDC confirmation for HIV-reactive blood donors in our center from 2014 to 2024 were analyzed systematically to confirm HIV infection and identify infection status. Results: There were 1 235 samples (0.13%, 1 235/928 000) reactive in HIV blood screening over the 11-year period. A-mong them, 199 donors (16.11%) in asymptomatic HIV infection (HIV Ag/Ab++&HIV RNA+), 2(0.16%) as acute early HIV infection (HIV Ag/Ab+-&HIV RNA+) and 7(0.57%) as window-period infection (HIV RNA positive only) were confirmed. Donors with the result of HIV Ag/Ab+-&HIV RNA-(single-positive) were all excluded for HIV infection, while 1 in 6 HIV Ag/Ab++&HIV RNA-donors (double-positive) was confirmed to have HIV infection. When HIV Ag/Ab reagents were used continuously before and after the follow-up, it's observed in one reagent that the proportion of negative results in subsequent follow-up in single-positive donors who had negative results in the first sampling was significantly higher than the proportion of negative results in subsequent follow-up in those initially single-positive (P<0.05). But no significant difference was observed in another reagent (P>0.05). When reagents were changed in follow-up, the rate of singlepositive donors with negative results in the first sampling reached 96.7%, which was significantly higher than the negative rate of those without reagent changing in follow-up (P<0.05). Conclusion: Based on the serological and nucleic acid testing results of HIV blood screening, the confirmation of HIV infection and identification of infection status can be achieved accurately and efficiently. All HIV Ag/Ab+-&HIV RNA-donors were confirmed as false positive, and should be maintained their eligibilities for blood donation, but recommended to pass the retest before next donation. Using a different reagent for retesting helps improve the eligible rate. HIV Ag/Ab++&HIV RNA-donors should be deferred permanently due to the risk of true positivity.

Objective: To explore the management of blood donors tested reactive to HCV in blood screening based on confirmation of HCV infection. Methods: Multiple HCV antibody assays, repeating HCV RNA testing, follow-up of blood donors and retesting of archive samples were performed to confirm HCV infection, identify infection status, and exclude false positives in blood donors reactive to HCV in blood screening. Results: From 2011 to 2024, the unqualified rate of HCV detection in blood screening was 2.45‰(2 751/1 122 026). Among these, anti-HCV+-&NAT-accounted for 1.85‰, followed by anti-HCV++ at 0.60‰. The proportion of anti-HCV+-&NAT-and HCV RNA yields was extremely low (0.007‰). The positive rate of anti-HCV+-&NAT-samples tested by electrochemiluminescence method (ELCIA) was approximately 7.5%, differing among reagents (P<0.05). The follow-up of anti-HCV+-&NAT-donors showed that 96.2% (202/210) were false positives, but 51.4% of donors remained anti-HCV+-&NAT-during follow-up. Among them, 8 donors (3.8%) could not be ruled out from HCV infection due to positive retesting by ELCIA. Of the anti-HCV+-&NAT-donors who were reactive at the first follow-up, 86.8% remained anti-HCV+-&NAT-at the second follow-up. The sampling confirmation data showed that all of 260 anti-HCV++ donors were confirmed as anti-HCV positive, and the proportion of false positives or missed detections by NAT was very low. Two occult HBV infections (OBIs) and one HBsAg carrier were identified among the 3 anti-HCV +-&NAT+ donors, and no HCV infection was confirmed in 5 anti-HCV--&HCV RNA + donors. Conclusion: The prevalence of HCV among blood donors in Dalian was about 0.06%, with extremely low proportion of window-period infection and slightly higher proportion of resolved infections than that of current infections. The majority of anti-HCV+-&NAT-were false positive. Blood donors confirmed as false positive should be qualified in blood screening 3 months later before next donation. In order to reduce the false positive results, it was advisable to avoid the same type of supplementary reagents as the initial reagents when performing confirmation.

Objective: To evaluate repeated testing on blood screening platforms in confirmation of non-discriminatory reactive (NDR) samples in nucleic acid testing (NAT). Methods: A total of 102 HBsAg-negative/NAT NDR samples were collected from voluntary blood donors at Dalian Blood Center between January 2021 and December 2023. Repeated testing was performed using two NAT platforms (Cobas s201 and Panther). For the first round of repeated testing, all samples were tested 12 times on each system; for the second round, the samples which were non-reactive or only reactive once in the first round were tested an additional 8 times. Anti-HBc and anti-HBs was detected using electrochemiluminescence assay (ECA). Meanwhile, blood donors were followed up. Results: The proportion of anti-HBc+ in 102 NDR samples was 88.2%. Forty-one samples (40.2%, 41/102) and 7 samples were confirmed HBV DNA+ in first-round and second-round repeated testing, respectively. The cumulative confirmation rate of HBV DNA+ was 47.1% (48/102) after repeated testing. Extra five blood donors detected HBV DNA+ in follow-up were identified as anti-HBc+ occult hepatitis B virus infection (OBI), while no window period infection was observed. Ultimately, there were 53 HBV infected donors confirmed, 46 HBV infection-unconfirmed, and 3 HBV uninfected. No significant difference was observed between the confirmation rate of the first-round testing and the cumulative confirmation rate after the second-round testing (P>0.05). The proportion of anti-HBc+ donors was quite high in both HBV infection-confirmed (98.1%) and unconfirmed group (82.6%), and donors with seronegative and anti-HBs-only occupied a high proportion in the latter (P<0.05). Conclusion: Numerous repeated testing of NDR samples using NAT platforms cannot achieve complete confirmation of HBV infection. Supplementary anti-HBc testing can minimize potential OBI risk among NDR donors, and is low-cost and efficient.

Objective: To investigate the impact of current laboratory testing models and screening strategies on blood safety by analyzing repeat nucleic acid testing (NAT) results obtained from blood donors who had dual-reactive enzyme-linked immunosorbent assay (ELISA) results but were initially nonreactive by NAT. Methods: Samples from blood donors with dual-reactive ELISA results and initially nonreactive NAT results collected at Dalian Blood Center from 2017 to 2025 were included. Repeated testing was performed using two NAT systems: transcription-mediated amplification (TMA) and polymerase chain reaction (PCR). The results were analyzed in terms of the frequency of reactive NAT results and the testing strategies, including minipool and individual donation testing. Results: A total of 199 samples with dual-reactive ELISA results and initially nonreactive NAT results were included，comprising 66 HBsAg-reactive samples，130 anti-HCV-reactive samples，and 3 HIV Ag/Ab-reactive samples. Among the HBsAg-reactive samples，57（57/66，86.4%）showed at least one reactive NAT result upon repeated testing，and 49 showed two or more repeat reactive results. Anti-HBc was reactive in 64 samples（64/66，97.0%）and nonreactive in 2 samples（2/66，3.0%）；both anti-HBc-nonreactive samples remained nonreactive in repeated NAT. No stable repeat nucleic acid reactivity was observed in anti-HCV- or HIV Ag/Abreactive samples. The TMA system detected 50 reactive samples（50/66，75.8%）through three combined assays plus one discriminatory assay, whereas the PCR system detected 51 reactive samples (51/66, 77.3%) through four rounds of individual-donation testing. Fortyfour samples were reactive on both systems, 6 were reactive only on the TMA system, and 7 were reactive only on the PCR system. Conclusion: Among blood donors with dual-reactive ELISA results and initially non-reactive NAT results, HBsAg-reactive samples contain a high proportion of low viral load HBV-related cases, with intermittent and probabilistic nucleic acid detectability. Both testing frequency and detection strategy significantly affect the detection of low viral load samples. These findings highlight the importance of recognizing the detection characteristics of low viral load HBV-related samples in order to optimize blood screening strategies and improve blood safety.

Objective: To establish quality standards for pooled cryoprecipitate based on measured data, addressing the issues of wide variability in coagulation factor content attributable to individual donor differences and imprecise clinical dosing with single-bag cryoprecipitate. Methods: A total of 120 bags of fresh frozen plasma of different ABO blood groups were enrolled. Single-bag cryoprecipitate was prepared using either the centrifugation method or the siphonage method. For each blood group and each preparation method, every five single units of the same blood group were pooled into one bag of pooled cryoprecipitate. Factor Ⅷ content, fibrinogen content, and volume were measured in source plasma, single-bag cryoprecipitate, and pooled cryoprecipitate. Recovery rates and coefficients of variation were calculated, and the effect of the pooling process on product homogeneity was analyzed. Results: The factor Ⅷ content, fibrinogen content, and volume of pooled cryoprecipitate were significantly higher than those of single-unit cryoprecipitate and were linearly correlated with the number of units pooled. After pooling, the coefficient of variation for factor Ⅷ content decreased from 34.9%-35.2% (single units) to 18.2%-20.7% (pools), and the coefficient of variation for fibrinogen content decreased from 27.0%-28.6% to 13.1%-13.4%. The pooling procedure had no significant effect on the recovery rates of factor Ⅷ or fibrinogen (P>0.05). Conclusion: For pooled cryoprecipitate prepared by pooling five single units, the proposed quality standards are: factor Ⅷ content≥400 IU/bag, fibrinogen content≥750 mg/bag, volume 200-250 mL/bag, and sterility test negative. These standards can provide a basis for large-scale preparation and clinical application of pooled cryoprecipitate.

BACKGROUND: Senescent human skin primary fibroblast (FB) models have been established for studying aging-related, proliferative, and inflammatory skin diseases. The aim of this study was to compare the transcriptome characteristics of human primary dermal FBs from children and the elderly with four senescence models. METHODS: Human skin primary FBs were obtained from healthy children (FB-C) and elderly donors (FB-E). Senescence models were generated by ultraviolet B irradiation (FB-UVB), D-galactose stimulation (FB-D-gal), atazanavir treatment (FB-ATV), and replication exhaustion induction (FB-P30). Flow cytometry, immunofluorescence staining, real-time quantitative polymerase chain reaction, co-culturing with immune cells, and bulk RNA sequencing were used for systematic comparisons of the models. RESULTS: In comparison with FB-C, FB-E showed elevated expression of senescence-related genes related to the skin barrier and extracellular matrix, proinflammatory factors, chemokines, oxidative stress, and complement factors. In comparison with FB-E, FB-UVB and FB-ATV showed higher levels of senescence and expression of the genes related to the senescence-associated secretory phenotype (SASP), and their shaped immune microenvironment highly facilitated the activation of downstream immune cells, including T cells, macrophages, and natural killer cells. FB-P30 was most similar to FB-E in terms of general transcriptome features, such as FB migration and proliferation, and aging-related characteristics. FB-D-gal showed the lowest expression levels of senescence-related genes. In comparisons with the single-cell RNA sequencing results, FB-E showed almost complete simulation of the transcriptional spectrum of FBs in elderly patients with atopic dermatitis, followed by FB-P30 and FB-UVB. FB-E and FB-P30 showed higher similarity with the FBs in keloids. CONCLUSIONS Each senescent FB model exhibited different characteristics. In addition to showing upregulated expression of natural senescence features, FB-UVB and FB-ATV showed high expression levels of senescence-related genes, including those involved in the SASP, and FB-P30 showed the greatest similarity with FB-E. However, D-galactose-stimulated FBs did not clearly present aging characteristics.
Humans ; Fibroblasts/drug effects* ; Cellular Senescence/physiology* ; Skin/metabolism* ; Child ; Transcriptome/genetics* ; Aged ; Ultraviolet Rays ; Cells, Cultured ; Galactose/pharmacology*

Alzheimer's disease (AD) is the most common cause of dementia and is a growing public health challenge. Neuroinflammation has been proposed as a prominent pathological feature of AD and has traditionally been attributed to the innate immune system. However, emerging evidence highlights the involvement of adaptive immunity, particularly T and B lymphocytes, in the neuroinflammatory processes of AD. It remains unclear how adaptive immune responses, originally intended to protect the body, contribute to chronic inflammation and neuronal dysfunction in AD. Here, we review the roles of adaptive immunity, cellular composition, and niches and their contribution to AD development and progression. Notably, we synthesize the crosstalk between adaptive immunity and the innate immune system of the central nervous system (CNS), which is mainly mediated by glial cells and myeloid cells, and their interrelationships with amyloid-β (Aβ)/Tau pathology. We hypothesized that the alterations observed in innate immunity in AD mirror age-related immune alterations, whereas the dysregulation of adaptive immunity contributes more accurately to disease-specific immune responses. Targeting adaptive immunity in the context of neuroinflammation may provide new insights into potential therapeutic strategies designed to modulate immune responses, thereby facilitating the diagnosis, intervention, and treatment of AD.
Alzheimer Disease/metabolism* ; Humans ; Adaptive Immunity/physiology* ; Immunity, Innate/immunology* ; Animals ; Neuroinflammatory Diseases/immunology* ; Inflammation/immunology* ; Amyloid beta-Peptides/metabolism*

OBJECTIVE To provide empirical basis for promoting the dynamic adjustment of the pharmaceutical care catalogue and the formulation of policies such as hierarchical payment of medical insurance. METHODS A multicenter cross- sectional survey method was adopted to conduct a questionnaire survey among 424 inpatients in 22 tertiary medical institutions in 12 prefecture-level cities of Hubei Province to evaluate their demand for pharmaceutical care， willingness to pay and preference for service forms. Combined with univariate and multivariate Logistic regression analysis， the influencing factors and key factors that affect patients’ willingness to pay for pharmaceutical care were identified. RESULTS Only 39.86% of the patients were aware of pharmaceutical care or pharmacists， and 89.62% of the patients hope to receive pharmaceutical care. Among the 16 types of pharmaceutical care， the patients surveyed had a relatively high recognition rate for guidance on drug usage and dosage， notification of medication precautions， and the identification， prevention and handling of adverse drug reactions. 96.70%， 95.30%， and 94.12% respectively expressed strong approval and approval. The demand for services such as insurance-related policy consultation， popular science on the mechanism of drug action， and assessment of the combined use of traditional Chinese and Western medicines was relatively low， with 61.65%， 68.47%， and 68.47% expressing strong approval and approval respectively. The positive influencing factors of willingness to pay were household monthly income ＞ 5 000 yuan （OR＝1.742）， awareness of pharmaceutical care or pharmacists （OR＝3.620）， and the desire to receive pharmaceutical care （OR＝4.686） （P＜0.05）， while self-rating health as “good” （OR＝0.390） was a negative influencing factor （P＜0.05）. Cardiovascular and cerebrovascular diseases （54.48%） and antihypertensive drugs （45.05%） were the service scenarios that the surveyed patients most hope to be covered. 85.14% of the patients preferred “service when xiaochnye@126.com needed”， with a single service duration of less than 10 minutes being appropriate （84.43%）， and the willingness to pay within 20 yuan being the main type （85.38%）. CONCLUSIONS Based on the characteristics of patients’ needs and payment behaviors， it is suggested that our country could consider establishing a hierarchical payment mechanism for pharmaceutical care， and focus on differentiated design in combination with diseases and medication situations. At the same time， the rights， responsibilities and service standards of resident pharmacists in the links such as medication reorganization and medical order review should be further clarified to comprehensively enhance the clinical value and policy operability of pharmaceutical care.