Transfusion and Anemia Expertise Initiative-Control/Avoidance of Bleeding developed a strategy for platelet and plasma infusion management in critically ill children based on systematic reviews and consensus meetings of international multidisciplinary experts. One good practice statement and six expert consensus statements were proposed for plasma and platelet transfusions in critically ill children following severe trauma, traumatic brain injury, and/or intracranial hemorrhage. This article introduces the specific methods and basis for the formation of recommendations in this part of the guide.

Objective: To investigate the current knowledge status of transfusion medicine among pediatricians/postgraduates and the reliability of large language models (LLMs) for assisted learning, and to assess changes in pediatricians' transfusion medicine knowledge before and after the implementation of the "Pediatric Transfusion Guideline" (hereafter referred to as the "Guideline"). Methods: In January 2022 (prior to the implementation of the "Guideline"), a questionnaire was developed based on the "Guideline" content and distributed to pediatricians. Subsequently, in July 2025 (after the implementation of the "Guideline"), the "Pediatric Transfusion Medicine Knowledge Questionnaire" was designed based on the content of the January 2022 questionnaire. This questionnaire survey was conducted on pediatricians/postgraduates and LLMs. We analyzed the level of transfusion medicine knowledge among pediatricians/postgraduates and the reliability of LLMs for assisted learning, and compared the accuracy of pediatricians' responses before and after "Guideline" implementation. Results: The survey results after the implementation of the "Guidelines" revealed that pediatricians/postgraduates achieved response accuracy rates exceeding 80% on the topic of "Patient Blood Management". However, response accuracy rates were below 30% for topics including "Types and Indications of Blood Components/Products" and "E-valuation of Transfusion Efficacy". The pediatricians' accuracy rates for related questions before and after the implementation of the "Guidelines" were 14.7%-68.9% and 3%-38%, respectively, and the comparison of accuracy rates for each question showed significant differences (P<0.001). The accuracy rates of the LLMs on the questionnaire were all below 90%. Among them, Doubao (81.1%) and Kimi (86.4%) achieved relatively higher accuracy rates, while Tencent Yuanbao (Hunyuan) had the lowest accuracy rate at only 59.5%. Conclusion: The implementation of the "Guideline" may have improved pediatricians' knowledge level of pediatric transfusion medicine. However, their knowledge level of pediatric transfusion remains low, and LLMs cannot yet provide absolutely reliable guidance. Systematic training in pediatric transfusion medicine is urgently needed.

[摘 要] 目的：构建负载STING激动剂DMXAA的锰卟啉金属有机框架纳米颗粒（DPM），探讨其对三阴性乳腺癌（TNBC）细胞4T1及其小鼠移植瘤的治疗效果。方法：通过物理吸附法制备 DPM 纳米颗粒，利用透射电镜、扫描电镜及纳米粒度电位仪表征其形貌与理化性质。常规培养4T1细胞，细胞实验分为对照组、超声辐照组（US组）、DPM治疗组（DPM组）和DPM治疗联合超声辐照组（DPM + US组），用CCK-8法检测细胞活性，免疫荧光法检测高迁移率族蛋白B1（HMGB1）和钙网蛋白（CRT）的表达，WB法检测STING通路相关蛋白的表达。构建4T1细胞移植瘤小鼠模型，分为四组，处理同细胞实验，测量肿瘤体积，免疫荧光法检测移植瘤组织中Ki-67、HMGB1、CRT和缺氧诱导因子-1ɑ（HIF-1ɑ）蛋白的表达，TUNEL法检测细胞凋亡，流式细胞术检测免疫细胞活化情况，对主要器官进行H-E染色，以评估纳米材料的体内安全性。结果：DPM呈梭形，平均粒径（268 ± 3.302）nm，电位（33.1 ± 0.87）mV。细胞实验中，DPM联合超声辐照可明显抑制4T1细胞的增殖（P < 0.001），提高4T1细胞中ROS水平（P < 0.001），诱导4T1细胞CRT表达上调（P < 0.001），HMGB1从细胞核中移至细胞质，激活STING信号通路[p-STING、p-TBK1、p-IRF3蛋白表达均显著增加（均P < 0.001）]。体内实验中，DPM联合超声辐照可显著抑制4T1细胞移植瘤生长（P < 0.001）并促进免疫细胞表型转化（P < 0.001），抑制移植瘤组织中Ki-67、HIF-1α蛋白表达（均P < 0.01），谷胱甘肽（GSH）产生（P < 0.01），促进CRT、HMGB1蛋白表达、ROS产生（P < 0.001），对主要器官结构无明显影响。结论： DPM联合超声辐照可通过激活STING通路显著抑制4T1细胞及其移植瘤的生长，诱导抗肿瘤免疫应答，且对主要器官无明显毒性。

ObjectiveTo develop a machine learning (ML)-based prediction model for neurological rehabilitation outcomes of stroke patients. MethodsA total of 420 stroke patients admitted to the Fuzhou First People's Hospital from October, 2022 to October, 2024 were enrolled as the training set. According to the modified Rankin Scale (mRS) scores three months after discharge, the patients were divided into prognosis group (n = 289) and poor prognosis group (n = 131). An additional 180 stroke patients hospitalized in the same hospital from November, 2024 to April, 2025 were selected as the validation set. Univariate analysis, least absolute shrinkage and selection operator regression, and multivariate logistic regression were performed to identify independent influencing factors for the prognosis of neurological function recovery. Using the screened independent influencing factors as feature variables, six ML models were established, including logistic regression, linear discriminant analysis, naive Bayes, support vector machine, random forest and extreme gradient boosting (XGBoost). The area under the receiver operating characteristic curve (AUC), confusion matrix indicators (accuracy, precision, recall and F1-score), calibration curve and decision curve analysis were adopted to evaluate the predictive efficacy, calibration degree and clinical net benefit of each model, with external validation conducted in the validation set. The SHapley Additive exPlanations framework was used to interpret the optimal model, and bar charts were applied to visualize the feature importance of the best model. ResultsAge, National Institutes of Health Stroke Scale (NIHSS) score, collateral circulation grading, fasting plasma glucose (FPG), lymphocyte percentage (LYMPH%), and homocysteine (Hcy) were independent risk factors for poor neurological rehabilitation prognosis (P < 0.05). For the XGBoost model, the AUC of the training and validation sets were 0.963 (95%CI 0.947 to 0.979) and 0.825 (95%CI 0.764 to 0.885), respectively, while the accuracy was 88.81% and 77.22%, the precision was 92.86% and 68.42%, the recall was 69.47% and 47.27%, and the F1-score was 79.48% and 55.91%, optimal in both calibration and clinical net benefit. The feature importance ranking for the XGBoost model from high to low was NIHSS score, age, collateral circulation grading, FPG, Hcy and LYMPH%. ConclusionThe interpretable XGBoost ML model exhibits excellent predictive efficacy and favorable clinical applicability in predicting neurological rehabilitation outcomes for stroke patients.

Objective:To investigate the expression and clinical significance of long noncoding RNA (lncRNA) AP006284.1 in bladder cancer tissues, and to analyze the regulatory effect and downstream mechanism of AP006284.1 knockdown on the proliferation and migration of bladder cancer cells. Methods:The expression of AP006284.1 in bladder cancer tissues, and the relationship between AP006284.1 expression and tumor stage and disease-free survival of bladder cancer patients were analyzed in the gene expression omnibus (GEO) database. AP006284.1 gene expression in bladder cancer cell lines, including MGH-U3, T24, UMUC-3, J82 and 5637, and bladder epithelial immortalized SV-HUC-1 cell were detected by real-time reverse transcription-PCR (RT-qPCR) assay. J82 cells were divided into the control group and the transfection group, and transfected with control plasmid and AP006284.1 knockdown plasmid, respectively. The effect of AP006284.1 knockdown on the proliferation of J82 cells was detected by RT-qPCR and cell counting kit-8 (CCK-8) assay. The effect of AP006284.1 knockdown on the migration of J82 cells was determined by the scratch healing assay. The target gene of AP006284.1 was predicted by LncRNA2Target and LncRNome databases. The target fragment of wild-type AP006284.1 ( AP006284.1-Wt) or mutant AP006284.1 ( AP006284.1-Mut) was constructed into pGL3 plasmid by dual luciferase gene reporter assay. J82 cells were co-transfected with miR-205-3p or miR-negtive control (miR-NC) to validate the targeting relationship between AP006284.1 and miR-205-3p. The correlation between miR-205-3p and AP006284.1 expression in bladder cancer tissues was further analyzed by the GEO database. The effect of AP006284.1 knockdown on the expression of miR-205-3p gene in J82 cells was detected by RT-qPCR assay. The effect of AP006284.1 knockdown on the expression of phosphorylated Ras protein (p-Ras), phosphorylated Raf protein (p-Raf), phosphorylated mitogen-activated protein kinase kinase (p-MEK), and phosphorylated extracellular signal-regulated kinase (p-ERK) of the ERK pathway was detected by Western blotting in J82 cells. Results:GEO database analysis showed that the relative expression of AP006284.1 in bladder cancer tissues ( n=304) was significantly higher than that in normal tissues ( n=28, P<0.01). The relative expression of AP006284.1 was positively correlated with the tumor stage of the bladder cancer patients ( P<0.01). Compared with bladder cancer patients with low expression of AP006284.1, patients with high expression had a lower disease-free survival ( P<0.01). Compared with the SV-HUC-1 cell (1.02±0.34), the expression level of AP006284.1 gene was upregulated in MGH-U3 cell (5.77±0.37), T24 cell (3.02±0.40), UMUC-3 cell (3.62±0.59), J82 cell (7.19±0.24) and 5637 cell (5.59±0.30) (all P<0.01). The expression level of the AP006284.1 gene was the highest in J82 cells, therefore, the J82 cells were selected for the study. The expression level of AP006284.1 gene in the control group (7.20±0.26) was 6.92 times higher than that in the transfection group (1.04±0.28, t=16.16, P<0.01). Compared with the control group (0.74±0.11, 1.35±0.09, 1.63±0.14, 1.74±0.11), the absorbance ( A) values of J82 cells in the transfection group (0.49±0.06, 0.95±0.14, 1.09±0.08, 1.13±0.11) were reduced than those in the control group at the 24, 36, 48 and 60 h after AP006284.1 knockdown (all P<0.05). The migration distance of J82 cells in the control group was significantly longer than that in the transfection group. The migration rate of the control group [(65.03±6.20)%], which was 2.58 times higher than that of the transfection group [(25.22±3.45)%, t=5.61, P<0.01]. The target site of miR-205-3p containing AP006284.1 was predicted by LncRNA2Target and LncRNome databases. Compared with miR-NC group (1.00±0.11), the relative activity of dual luciferase of AP006284.1-Wt gene was significantly downregulated in the miR-205-3p group (0.31±0.07, t=5.47, P<0.01). Compared with the miR-NC group (0.97±0.14), the relative activity of dual luciferase of AP006284.1-Mut vector (0.98±0.07) was not significantly change ( t=0.09, P>0.05). The GEO database analysis showed that the expression of AP006284.1 in bladder cancer tissues was negatively correlated with the expression of miR-205-3p ( P<0.01). The expression level of miR-205-3p gene in the transfection group (5.42±0.24) was 5.21 times higher than that in the control group (1.04±0.40, t=9.40, P<0.01). Compared with the control group (3.22±0.17, 5.56±0.19, 4.38±0.17, 5.74±0.36), the expressions of p-Ras (2.33±0.12), p-Raf (1.61±0.20), p-MEK (1.57±0.25), and p-ERK (2.40±0.28) of the ERK pathway were decreased in the transfected J82 cells (all P<0.01). Conclusions:AP006284.1 is highly expressed in bladder cancer tissues. Knockdown of AP006284.1 can inhibit the proliferation and migration of bladder cancer cells by regulating the miR-205-3p and ERK pathway proteins.

This study aims to explore whether Naoxintong Capsules improve multiple cerebral infarctions and myocardial injury via promoting angiogenesis, thereby exerting a simultaneous treatment effect on both the brain and heart. Male SD rats were randomly divided into six groups: sham-operated group, model group, high-dose, medium-dose, and low-dose groups of Naoxintong Capsules(440, 220, and 110 mg·kg~(-1)), and nimodipine group(10.8 mg·kg~(-1)). Rat models of multiple cerebral infarctions were established by injecting autologous thrombus, and samples were collected and tested seven days after modeling. Evaluations included multiple cerebral infarction model assessments, neurological function scores, grip strength tests, and rotarod tests, so as to evaluate neuromotor functions. Morphological structures of brain and heart tissue were observed using hematoxylin-eosin(HE) staining, Nissl staining, and Masson staining. Network pharmacology was employed to screen the mechanisms of Naoxintong Capsules in improving multiple cerebral infarctions and myocardial injury. Neuronal and myocardial cell ultrastructures were observed using transmission electron microscopy. Apoptosis rate in brain neuronal cells was detected by TdT-mediated dUTP nick end labeling(TUNEL) staining, and reactive oxygen species(ROS) levels in myocardial cells were measured. Immunofluorescence was used to detect the expression of platelet endothelial cell adhesion molecule-1(CD31), antigen identified by monoclonal antibody Ki67(Ki67), hematopoietic progenitor cell antigen CD34(CD34), and hypoxia inducible factor-1α(HIF-1α) in brain and myocardial tissue. Western blot, and real-time quantitative polymerase chain reaction(RT-qPCR) were used to detect the expression of HIF-1α, vascular endothelial growth factor(VEGF), vascular endothelial growth factor receptor 2(VEGFR2), sarcoma(Src), basic fibroblast growth factor(bFGF), angiopoietin-1(Ang-1), and TEK receptor tyrosine kinase(Tie-2). Compared with the model group, the medium-dose group of Naoxintong Capsules showed significantly lower neurological function scores, increased grip strength, and prolonged time on the rotarod. Pathological damage in brain and heart tissue was reduced, with increased and more orderly arranged mitochondria in neurons and cardiomyocytes. Apoptosis in brain neuronal cells was decreased, and ROS levels in cardiomyocytes were reduced. The microvascular density and endothelial cells of new blood vessels in brain and heart tissue increased, with increased overlapping regions of CD31 and Ki67 expression. The relative protein and mRNA expression levels of HIF-1α, VEGF, VEGFR2, Src, Ang-1, Tie-2, and bFGF were elevated in brain tissue and myocardial tissue. Naoxintong Capsules may improve multiple cerebral infarctions and myocardial injury by mediating HIF-1α/VEGF expression to promote angiogenesis.
Animals ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Rats, Sprague-Dawley ; Rats ; Cerebral Infarction/genetics* ; Hypoxia-Inducible Factor 1, alpha Subunit/genetics* ; Vascular Endothelial Growth Factor A/genetics* ; Capsules ; Signal Transduction/drug effects* ; Humans ; Brain/metabolism* ; Myocardium/metabolism* ; Apoptosis/drug effects*

Real-world study (RWS), based on multi-source data from real medical environments, is gradually becoming an important supplement to traditional randomized controlled trials, and its application in the field of transfusion medicine is becoming increasingly widespread. This article systematically reviews the definition and methodological system of RWS, examines its application cases in clinical blood transfusion research, and discusses the advantages, limitations, and future research directions of RWS, aiming to provide a reference for evidence-based research in blood transfusion medicine.

Growth arrest DNA damage-inducible protein 45 β (GADD45B) has been reported to be a regulatory factor for active DNA demethylation and is implicated in the modulation of synaptic plasticity and chronic stress-related psychopathological processes. However, its precise role and mechanism of action in stress susceptibility remain elusive. In this study, we found a significant reduction in GADD45B expression specifically in the ventral, but not the dorsal hippocampal CA1 (dCA1) of stress-susceptible mice. Furthermore, we demonstrated that GADD45B negatively regulates susceptibility to social stress and NMDA receptor-dependent long-term potentiation (LTP) in the ventral hippocampal CA1 (vCA1). Importantly, through pharmacological inhibition using the NMDA receptor antagonist MK801, we provided further evidence supporting the hypothesis that GADD45B potentially modulates susceptibility to social stress by influencing NMDA receptor-mediated LTP. Collectively, these results suggested that modulation of NMDA receptor-mediated synaptic plasticity is a pivotal mechanism underlying the regulation of susceptibility to social stress by GADD45B.
Animals ; Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors* ; CA1 Region, Hippocampal/drug effects* ; Male ; Stress, Psychological/physiopathology* ; Mice ; Neuronal Plasticity/drug effects* ; Long-Term Potentiation/drug effects* ; Mice, Inbred C57BL ; Antigens, Differentiation/metabolism* ; Dizocilpine Maleate/pharmacology* ; Excitatory Amino Acid Antagonists/pharmacology* ; GADD45 Proteins

OBJECTIVE: This study investigates the efficacy and mechanisms of Qingjie Fuzheng Granules (QFG) in inhibiting colitis-associated colorectal cancer (CAC) development via RNA sequencing (RNA-seq) and 16S ribosomal RNA (rRNA) correlation analysis. METHODS: CAC was induced in BALB/c mice using azoxymethane (AOM) and dextran sulfate sodium (DSS), and QFG was administered orally to the treatment group. The effects of QFG on CAC were evaluated using disease index, histology, and serum T-cell ratios. RNA-seq and 16S rRNA analysis assessed the transcriptome and microbiome change. Key pharmacodynamic pathways were identified by integrating these data and confirmed via Western blotting and immunofluorescence. The link between microbiota and CAC-related markers was explored using linear discriminant analysis effect size and Spearman correlation analysis. RESULTS: Long-term treatment with QFG prevented AOM/DSS-induced CAC formation, reduced levels of interleukin (IL)-1β, tumor necrosis factor-alpha (TNF-α), IL-6, and interferon γ (IFN-γ), and increased CD3⁺ and CD4⁺/CD8⁺ T cells ratio, without causing hepatic or renal toxicity. A 16S rRNA analysis revealed that QFG rebalanced the Firmicutes/Bacteroidetes ratio and mitigated AOM/DSS-induced microbiota disturbances. Transcriptomics and Western blotting analysis identified the nucleotide-binding oligomerization domain-containing protein 2 (NOD2)/nuclear factor kappa-B (NF-κB) pathway as key for QFG's treatment against CAC. Furthermore, QFG decreased the abundance of Bacilli, Bacillales, Staphylococcaceae, Staphylococcus, Lactobacillales, Aerococcus, Alloprevotella, and Akkermansia, while increasing Clostridiales, Lachnospiraceae, Lachnospiraceae_NK4A136_group, Ruminococcaceae, and Muribaculaceae, which were highly correlated with CAC-related markers or NOD2/NF-κB pathway. CONCLUSION By mapping the relationships between CAC, immune responses, microbiota, and key pathways, this study clarifies the mechanism of QFG in inhibiting CAC, highlighting its potential for clinical use as preventive therapy.

To guide transfusion practice in critically ill children who often need plasma and platelet transfusions, the Transfusion and Anemia Expertise Initiative-Control/Avoidance of Bleeding (TAXI-CAB) developed Recommendations and Expert Consensus for Plasma and Platelet Transfusion Practice in Critically Ill Children. This guideline addresses 53 recommendations related to plasma and platelet transfusion in critically ill children with 8 kinds of diseases, laboratory testing, selection/treatment of plasma and platelet components, and research priorities. This paper introduces the specific methods and results of the recommendation formation of the guideline.