Objective: To investigate the damaging effects of red blood cell supernatant (RBC-S) stored for different durations (7 d, 14 d, and 28 d) on vascular endothelial cells, and to explore the underlying mechanisms using bioinformatics analysis, so as to provide references for optimizing red blood cell transfusion strategies. Methods: Human umbilical vein endothelial cells (HUVECs) were co-cultured with RBC-S stored for 7, 14 and 28 days, designated as the 7 d group, 14 d group and 28 d group respectively, which were collectively defined as the experimental groups. Cell damage was evaluated by cell proliferation assay (Cell Counting Kit8, CCK8), lactate dehydrogenase (LDH) release assay, 4′, 6diamidino2phenylindole (DAPI) staining, and flow cytometry for apoptosis and reactive oxygen species (ROS) levels. The damage degree of RBC-S on vascular endothelial cells was assessed by statistical analysis of damage data among different groups. Since the damage effect reached a plateau at all time points, the 28 d storage group was selected as the representative for further mechanistic studies. Transcriptomic analysis was performed to explore the role of frizzled class receptor 1 (FZD1) and Wnt signaling pathway in red blood cell storagerelated endothelial dysfunction. Results: Compared with the control group, the storage groups treated with 7 d, 14 d, and 28 d RBC-S showed significantly decreased cell proliferation rates [control group 100%, 7 d group (69.51±2.30)%, 14 d group (74.54±2.89)%, 28 d group (73.59±2.36)%, P<0.05], significantly reduced numbers of DAPI-stained cell nuclei [control group (213±12.5) per field, 7 d group (140.33±17.04) per field, 14 d group (152.00±23.72) per field, 28 d group (144.33±19.09) per field, P<0.05] and significantly increased LDH release [control group (1), 7 d group (8.33±1.41), 14 d group (9.23±0.83), 28 d group (9.16±0.60), P<0.05]. There was no significant difference in the degree of damage caused by RBC-S among different storage groups (P>0.05). With the prolongation of storage time, free hemoglobin (FHb) gradually increased [control group (not detected), 7 d (16.57±6.38) mg/L, 14 d (76.80±22.83) mg/L, 28 d (286.97±29.02) mg/L, P<0.05]. The apoptotic rate (20.53±2.94)% and ROS relative intensity (5.13±0.91) in the 28 d storage group were significantly higher than those in the control group (P<0.05). Transcriptomic analysis showed that FZD1 played a key role in vascular endothelial dysfunction induced by red blood cell storage and was closely related to the Wnt signaling regulatory network. Conclusion: RBC-S stored for 7 d, 14 d, or 28 d can all significantly damage vascular endothelial cells, and the damaging effect reaches a plateau at 7 d of storage. Mechanistic investigation of the 28 d group indicated that the downregulation of the FZD1/Wnt signaling pathway may play a critical role in vascular endothelial dysfunction induced by red blood cell storage, providing a theoretical basis for further optimizing red blood cell storage and transfusion strategies.

Persistent and maladaptive drug-related memories represent a key component in drug addiction. Converging evidence from both preclinical and clinical studies has demonstrated the potential efficacy of the memory reconsolidation updating procedure (MRUP), a non-pharmacological strategy intertwining two distinct memory processes: reconsolidation and extinction-alternatively termed "the memory retrieval-extinction procedure". This procedure presents a promising approach to attenuate, if not erase, entrenched drug memories and prevent relapse. The present review delineates the applications, molecular underpinnings, and operational boundaries of MRUP in the context of various forms of substance dependence. Furthermore, we critically examine the methodological limitations of MRUP, postulating potential refinement to optimize its therapeutic efficacy. In addition, we also look at the potential integration of MRUP and neurostimulation treatments in the domain of substance addiction. Overall, existing studies underscore the significant potential of MRUP, suggesting that interventions predicated on it could herald a promising avenue to enhance clinical outcomes in substance addiction therapy.
Humans ; Substance-Related Disorders/psychology* ; Memory Consolidation/physiology* ; Animals ; Extinction, Psychological/physiology*

OBJECTIVES: To investigate the effect of pachymic acid on brown/beige adipocyte differentiation and lipid metabolism in preadipocytes. METHODS: 3T3-L1 MBX cells were induced to differentiate into beige adipocytes using a brown cocktail method. The impact of pachymic acid on the viability of 3T3-L1 MBX cells was evaluated using the CCK-8 assay. The formation of lipid droplets following treatment with pachymic acid was observed by oil red O staining. The mRNA expression levels of key browning genes, including uncoupling protein (Ucp) 1, the peroxisome proliferator activated receptor-γ coactivator (Pgc)-1α, and the PR domain-containing protein 16 (Prdm16), as well as the mRNA expression of sterol regulatory element-binding protein (Srebp) 1c, acetyl-coA carboxylase (Acc), fatty acid synthase (Fas), and hormone-sensitive triglyceride lipase (Hsl), adipose triglyceride lipase (Atgl), and carnitine palmitoyltransferase (Cpt) 1 were detected by quantitative reverse transcription polymerase chain reaction. The protein expression of Ucp1, Pgc-1a, and Prdm16 was detected by Western blotting. RESULTS: The 3T3-L1 MBX cells were induced in vitro to form beige adipocytes with high expression of key browning genes(Ucp1, Pgc-1α, and Prdm16), and beige adipose-marker genes (Cd137, Tbx1, and Tmem26). Concentrations range of 0-80 μmol/L pachymic acid were non-cytotoxic to 3T3-L1 MBX cells. Pachymic acid treatment significantly inhibited the differentiation of 3T3-L1 MBX cells, resulting in a notable decrease in lipid accumulation. There was a marked increase in the expression of key browning genes and their proteins products, such as Ucp1, Pgc-1α, and Prdm16, while the expressions of fat synthesis-related genes Srebp1c, Acc and Fas were significantly decreased (all P<0.05). The expressions of lipolysis-related genes (Hsl, Atgl, and Cpt1) were significantly increased (all P<0.05). Treatment with 20 μmol/L pachymic acid showed the most pronounced effect. CONCLUSIONS Pachymic acid can inhibit fat synthesis and promote lipid decomposition by regulating the brown formation and lipid differentiation of preadipocytes.
Animals ; Lipid Metabolism/drug effects* ; Mice ; Cell Differentiation/drug effects* ; Adipocytes, Beige/drug effects* ; 3T3-L1 Cells ; Adipocytes, Brown/drug effects* ; Triterpenes/pharmacology* ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Uncoupling Protein 1 ; Sterol Regulatory Element Binding Protein 1/metabolism*

[This corrects the article DOI: 10.1016/j.apsb.2022.03.002.].

Objective:To investigate the pattern of Mesothelin(MSLN)-specific T-cell(CTLs)immune reconstitution after allogeneic hematopoietic stem cell transplantation.Methods:Two cases of Mesothelin-positive patients with acute myeloid leukemia(AML)were screened,and the number of MSLN-CTLs and its subpopulations,the expression of surface PD-1/CTLA-4/TIM-3 im-mune exhaustion molecules,and the functions of secreted cytokines TNF-α and IFN-γ were monitored by flow cytometry after trans-plantation.They were compared with WT1-CTLs and cytomegalovirus(CMV)CTLs.Meanwhile,the relationship between CTL recon-stitution and Minimal residual disease(MRD)and leukemia recurrence was analyzed.Results:After transplantation,MSLN-CTLs,WT1-CTLs and CMV-CTLs can be detected,and intracellular factors were secreted.The phenotypes of WT1 specific CD8+T cells,MSLN specific CD8+T cells and CMV specific CD8+T cells were mainly TEM subsets and TEMRA subsets,and the TEM subsets of CMV specific CD8+T cells were more obvious.Compared with CMV-CTLs,the proportions of T Naive,TCM and TEMRA subsets were relatively higher in MSLN-specific CD8+T cells and WT1-specific CD8+T cells,and the expression levels of PD-1,CTLA-4 and TIM-3 were higher in MSLN-CTLs and WT1-CTLs.The dynamic changes of MSLN-CTLs and WT1-CTLs after transplantation were related to leukemia load and the chimerism rate of donor and recipient.Conclusion:After allogeneic hematopoietic stem cell transplantation,im-mune recovery to MSLN is found,which is different from WT1-CTLs and CMV-CTLs.

Objective:To analyze the application of situational teaching method based on the OBE concept in the teaching of Emergency and Critical Care Nursing course for undergraduate nursing students,and evaluate its effect on their clinical practice and internship adaptability.Methods:The third-year nursing students from 2018 to 2021 in a university in Anhui Province were selected as the research objects.Thirty students were selected as the experimental group each academic year,and taught using situational teaching method based on the OBE concept.Another 30 students from the same academic year were selected as the control group and taught using the conventional teaching method.The course teaching evaluation and course assessment scores of the two groups were compared,and the clinical practice ability and internship adaptability were compared before and after 3 months of hospital internship.Results:There was no significant difference in the course teaching evaluation and course assessment scores between the two groups(P＞0.05).The case analysis score of the experimental group was higher than that of the control group(P＜0.01).The scores of each dimension and total score of the clinical practice ability scale after internship in the observation group were higher than those in the control group(P＜0.05),and the scores of each dimension and total score of the internship adaptability scale were also higher than those in the control group(P＜0.05).Conclusion:Based on the concept of OBE,the situational teaching method is beneficial to improving the ability of case analysis,and enhancing the clinical practice ability and internship adaptability of nursing students.

OBJECTIVE To analyze the investigation and handling process of a suspected outbreak of hospital-ac-quired Serratia marcescens infection in the Neonatal Intensive Care Unit(NICU)of a certain hospital,and to pro-vide valuable reference for the prevention and control of hospital-acquired infections.METHODS An epidemiologi-cal investigation was conducted on three S.marcescens-positive neonates admitted to the NICU of the Affiliated Hospital of Guizhou Medical University from Oct.12 to 18,2024.Environmental hygiene sampling was carried out on the surfaces of the ward.The homology of the strains was analyzed,and effective intervention measures were taken.RESULTS The incidence of S.marcescens sepsis in the NICU from Oct.12 to 18,2024 was 2.20％(2/91)compared to 2.35％in the same period of 2023(2/85),no significant difference was found between the two time periods(P=0.946).Among the three S.marcescens-positive NICU neonates,one was considered com-munity-acquired,while the other two were diagnosed with S.marcescens sepsis.A total of 59 environmental hy-giene samples were collected,with one sample detecting S.marcescens in the bedside infusion pump slot of case 1.The homology analysis results showed high homology among the four S.marcescens strains but not completely homologous.After effective preventive and control measures were implemented,suspected S.marcescens cluste-ring events didn't further occurred.CONCLUSION The occurrence of this incident may be attributed to inadequate cleaning and disinfection of environmental surfaces and improper hand hygiene practices by some medical staff.

Objective:To construct and validate the risk prediction model of osteoporosis (OP) in the middle-aged and elderly healthy physical examination population.Methods:In this cross-sectional study, 18 030 middle-aged and elderly people with bone mineral density tested in Health Management Center of Guangxi Zhuang Autonomous Region Hospital from January 2020 to December 2022 were selected. The general data, physical examination index and biochemical blood index were collected. The subjects were divided into training set (12 621 cases) and validation set (5 409 cases) in a ratio of 7∶3 with the simple random sampling method. The variables were screened with minimum LASSO regression and logistic regression and the corresponding nomogram prediction model for the risk of osteoporosis in the middle-aged and elderly health examination population was established. The performance of the nomogram model was evaluated with the area under the receiver operating characteristic curve (ROC AUC), specificity, sensitivity, calibration curve (CAL), and decision curve (DCA).Results:The results of LASSO regression and multivariate logistic regression in training set showed that gender, age, body mass index, hip circumference, waist circumference, systolic blood pressure, total cholesterol, glutamyl transpeptidase and albumin/globulin ratio were the independent best predictors of OP risk in the middle-aged and elderly health examination population (all P<0.05). The ROC AUC-value of the training set was 0.895 (95% CI: 0.886-0.904), with a sensitivity of 87.25% and a specificity of 85.01%. The ROC AUC value of the validation set was 0.892 (95% CI: 0.886-0.898), with a sensitivity of 83.74% and a specificity of 82.46%. The CAL showed a C-index value of 0.790 in the training set and a C-index value of 0.784 in validation set. The CALs all showed deviation correction and obvious curves similar to the ideal line. DCA showed that when the OP risk threshold probability of the training set was 45%-93%, and the OP risk threshold probability of the validation set was 45%-92%, the nomogram model had better efficacy in predicting OP risk in the middle-aged and elderly physical examination population, and the two results were still relatively consistent. Both CAL and DCA showed good performance. Conclusion:This study establishes a practical prediction model for osteoporosis risk in the middle-aged and elderly population, it can provide an early warning for the timely detection of OP risk for the middle-aged and elderly people.

Objective:To establish a modified one-stage clotting assay (mOSA) based on the STA-R Evolution coagulation analyzer for quantifying emicizumab (EMI) concentration and to preliminarily evaluate its analytical performance; meanwhile to explore the clinical utility of the standard one-stage clotting assay (sOSA) in indirectly predicting EMI levels through surrogate factor Ⅷ (FⅧ) activity.Methods:A total of 30 pediatric patients with hemophilia A (HA) treated with EMI in the Hemophilia Treatment Center of Shenzhen Children′s Hospital from January 2023 to March 2025 were enrolled, and 48 post-treatment plasma samples were collected. EMI standards (2.5~100 μg/ml) were prepared using FⅧ-deficient plasma to establish the mOSA detection system. The linearity, accuracy, and precision of the method were evaluated. Surrogate FⅧ activity was measured by sOSA to estimate EMI concentrations, and its correlation with mOSA-derived EMI concentrations was analyzed using Spearman correlation analysis. The equivalent FⅧ activity in patient plasma samples was measured using a human chromogenic substrate assay-based FⅧ activity detection reagent, and Spearman correlation analysis was employed to evaluate its correlations with both the EMI concentrations measured by the mOSA method and estimated by the sOSA method respectively.Results:The established mOSA method for EMI detection showed excellent linearity in the range of 2.5?100 μg/ml ( Y=1.047 X?1.033, R 2=0.995, P<0.001). Average spike recovery rates at 25, 50, and 75 μg/ml were 101.55%(25.39/25.00), 105.31%(52.66/50.00), and 98.20%(73.65/75.00), respectively. Coefficients of variations of within-and inter-batch were 3.47%?4.80% and 6.30%?8.96%, respectively. A prediction model for EMI concentration was established as follows: estimated EMI concentration (μg/ml)=0.095×[alternative FⅧ activity (%) measured by sOSA]+2.652 ( R2=0.999, P<0.001). Validation demonstrated a strong correlation between the EMI concentration measured by the mOSA method and the EMI concentration estimated by the sOSA method ( r=0.989, P<0.001), with good consistency ( Y=1.014 X+0.684, R2=0.972, P<0.001). Both the EMI concentration measured by the mOSA method and the EMI concentration estimated by the sOSA method showed extremely strong correlations with the equivalent FⅧ activity ( r=0.986 and 0.987, respectively; P<0.001 for both). Conclusions:The mOSA system established on the STA-R Evolution analyzer demonstrates robust linearity, accuracy, and reproducibility, fulfilling clinical requirements for therapeutic drug monitoring of EMI. The sOSA method provides reliable indirect estimation of EMI concentrations through surrogate FⅧ activity, offering critical support for emergency decision-making.

Objective: To investigate the epidemiological characteristics of epidemic encephalitis B in Huzhou City, Zhejiang Province from 2007 to 2023, so as to provide the evidence for formulating prevention and control measures for epidemic encephalitis B. Methods: Epidemic encephalitis B case data in Huzhou City from 2007 to 2023 were collected through the National Notifiable Disease Reporting System of the Chinese Disease Prevention and Control Information System. The temporal, regional and population distribution characteristics of laboratory-confirmed epidemic encephalitis B cases were analyzed using the descriptively epidemiological method. Results: A total of 49 laboratory-confirmed epidemic encephalitis B cases were reported in Huzhou City from 2007 to 2023, and the average annual incidence was 0.10/105, showing a significant downward trend (P<0.05). The epidemic encephalitis B cases were concentrated from July to August, and July was the peak month, with 40 cases accounting for 81.63%. There was a statistically significant difference in the average annual incidences of epidemic encephalitis B among counties (districts) (P<0.05). Nanxun District had the highest reported incidence, with an average annual incidence of 0.23/105. There were 30 male cases and 19 female cases, with a male-to-female ratio of 1.58∶1. The youngest case was 5 months old, and the oldest was 49 years old. The children under 15 years were in the majority, with 42 cases accounting for 85.71%. Most of the cases were scattered children, with 25 cases accounting for 51.02%. There were 22 cases with no vaccination history and 21 cases with an unknown vaccination history, accounting for 44.90% and 42.86% respectively. All cases presented with fever. Other main clinical symptoms included listlessness, drowsiness, vomiting and headache, with 47, 40, 33 and 29 cases respectively, accounting for 95.92%, 81.63%, 67.35% and 59.18%, respectively. Conclusions The incidence of epidemic encephalitis B in Huzhou City remained at a relatively low level from 2007 to 2023, with Nanxun District being the high-risk area and July being the peak month for disease incidence. Fever and listlessness were the predominant clinical manifestations. Strengthening vaccination for children under 15 years should be prioritized.