ObjectiveThe malaria parasites remodel the host erythrocyte structure by exporting parasite proteins that interact with the membrane skeleton proteins of red blood cells (RBCs), facilitating their intracellular survival and pathogenicity. Skeleton-binding protein 1 (SBP1) is a conserved exported protein across Plasmodium species. In Plasmodium falciparum, SBP1 has been reported to interact with erythrocyte membrane skeleton proteins 4.1R and spectrin, while its contribution to erythrocyte remodeling and parasite virulence in Plasmodium berghei (Pb) remains unclear. This study aims to determine whether PbSBP1 associates with the host cytoskeletal protein 4.1R and to investigate its role in the remodeling of host RBCs and the pathogenicity of Plasmodium berghei. MethodsIn Plasmodium berghei, the relationship between PbSBP1 and the erythrocyte cytoskeletal protein 4.1R was examined using co-immunoprecipitation. A Pbsbp1 gene knockout mutant of Plasmodium berghei (Pbsbp1∆) was generated based on the principle of double crossover homologous recombination. The deformability of erythrocytes infected with Pbsbp1∆ parasites was assessed using microfluidic methods. Microchannels with an array of cylindrical pillars were used to detect modifications in infected RBC deformability. The infected RBCs were squashed between the rows and recovered between the columns and the transit velocity (μm/s) of infected RBCs travelling through the microchannel was recorded. The component of the erythrocyte membrane skeleton junctional complex, tropomodulin (TMOD), was fluorescently labeled, and the cytoskeletal network of infected erythrocytes was imaged using super-resolution stochastic optical reconstruction microscopy (STORM) to analyze ultrastructural changes in the cytoskeleton of wild-type (WT) and Pbsbp1∆-infected erythrocytes. Actin-based junctional complexes were displayed as individual clusters by the labeled TMOD in the STORM images, and the cluster densities and distances between adjacent clusters of infected RBCs were calculated. Additionally, rodent malaria models (BALB/c mice) and experimental cerebral malaria models (C57BL/6 mice) were employed to monitor the growth of Pbsbp1∆ and WT parasites during the intraerythrocytic stage and their capacity to induce cerebral malaria in mice. ResultsPbSBP1 may participate in the remodeling of infected erythrocytes through direct or indirect interaction with the erythrocyte cytoskeletal protein 4.1R. Microfluidic assays revealed that the deformability of erythrocytes infected with Pbsbp1∆ parasites was significantly enhanced compared to those infected with WT parasites. STORM imaging further demonstrated that the ultrastructure of the erythrocyte cytoskeleton in Pbsbp1∆-infected cells was altered relative to that in WT-infected erythrocytes. The distances between nearest neighbors of clusters had a tendency to increase while the cluster densities were decreased in Pbsbp1∆-infected RBCs compared to WT-infected RBCs. Subsequent phenotypic analysis indicated that the growth rate of Pbsbp1∆ parasites during the intraerythrocytic stage was significantly slower than that of WT parasites, and their ability to induce cerebral malaria in mice was also attenuated. These findings suggest that PbSBP1 is involved in the remodeling of the erythrocyte membrane skeleton, likely through its direct or indirect interaction with protein 4.1R, thereby regulating the deformability of infected erythrocytes and influencing the pathogenicity of the blood-stage parasites. ConclusionThis study establishes a role for PbSBP1 in host erythrocyte remodeling and parasite virulence, providing new research strategies for the prevention and treatment of malaria.

Aim To explore the effects of D-limonene on the steatosis of primary mouse hepatocytes and its potential mechanism of action.Methods Oleic acid-induced steatosis in primary mouse hepatocytes was used as a model to observe the effects of D-limonene on cell viability,cellular lipid content,and intracellular expression of proteins such as AMP-activated protein kinase(AMPK),acetyl-coenzyme A carboxylase 1(ACC1),and carnitine palmitoyl transferase 1A(CPT1A).Results It was found that a low dose of D-limonene could effectively enhance the viability of primary mouse hepatocytes.When oleic acid at a con-centration of 300 μmol·L-1 successfully induced steatosis in primary mouse hepatocytes,D-limonene re-duced the lipid content of the cells,and D-limonene up-regulated the cellular AMPK expression level,down-regulated the cellular ACC1 and fatty acid synthetase(FAS)expression levels,which in turn promoted the overexpression of CPT1A.Conclusions D-limonene has the effect of reducing lipid deposition in primary mouse hepatocytes,and the mechanisms may be related to the activation of AMPK,the inhibitions of ACC1 and FAS,and the up-regulation of CPT1A protein expres-sion level.

The receptor-binding region(RBD)of the spike protein(S)on the surface of porcine epi-demic diarrhea virus(PEDV)is a critical structural domain mediating viral invasion of host cells and serves as a key target for inducing neutralizing antibodies.In order to prepare antibodies that can be used to study the biological function of PEDV RBD and develop novel diagnostic and thera-peutic methods,recombinant RBD protein expressed in Sf9 insect cells was utilized as an immuno-gen to immunize BALB/c mice.Monoclonal antibodies(mAbs)were generated via hybridoma tech-nology,and positive hybridoma clones were screened using indirect ELISA.The reactivity of the mAbs was subsequently characterized.The results of ELISA,Western blot,and indirect immuno-fluorescence assay(IFA)showed that three monoclonal antibodies screened(3E5,4F9 and 5A5)had good reactivity with the virus and RBD protein.Antibody subtype results showed that 3E5 and 4F9 were of IgG1 subtypes and 5A5 was of IgM subtype.Neutralization assay further revealed that 3E5 monoclonal antibody had viral neutralizing activity.In this study,three monoclonal antibodies against PEDV RBD proteins were successfully prepared,providing the basis for the study of the bi-ological function of RBD proteins,PEDV serologic detection and vaccine development.

Pain modulation encompasses a complex neurobiological process, in which the lateral habenula (LHb) plays a crucial role in integrating, regulating and modulating pain signals. It is also involved in pain-related memory functions associated with perception, transmission and regulation of pain. Furthermore, the LHb collaborates with structures such as the spinal dorsal horn, forebrain, and amygdala to form an essential neural circuit that contributes to sensitization, development of tolerance, and adaptation processes related to pain. However, there remains limited understanding regarding the specific roles and interactions among different neuron subtypes within the LHb concerning pain regulation. Additionally, further investigation is warranted to explore functional changes and plasticity within both the LHb and its associated neural circuits in chronic pain models. Future research endeavors should utilize advanced neuroimaging techniques alongside optogenetics and gene editing technologies to elucidate intricate neural circuits, cellular architecture, and molecular mechanisms governing LHb function in pain regulation. In conclusion, this paper aims to comprehensively review existing literature on the involvement of the LHb and its neural circuits in modulating pain, thereby enhancing our understanding of their neurobiological mechanisms while providing novel targets for precise therapeutic strategies aimed at alleviating pain.

Objective:To analyze the predictive role of WT1 expression levels pre-and early post-transplantation on relapse and overall survival(OS)in patients with acute myeloid leukemia(AML)undergoing allogeneic hematopoietic stem cell transplantation(allo-HSCT)during their first complete remission(CR1).Methods:A retrospective analysis was conducted on the clinical data of 107 adult AML patients who underwent allo-HSCT during their CR1 at our center between May 2012 and December 2021.The predictive role of bone marrow WT1 expression levels before transplantation and at 3 and 6 months post-transplantation on relapse and OS was explored in combination with relevant clinical factors.Results:The median follow-up time for the 107 patients was 70(range:11-117)months.Among the patients,15 cases died.Kaplan-Meier survial analysis showed that the 3-year overall survival(OS)rate was 85.0％.20 patients experienced relapse,with a median time to relapse of 8(range:0.5-44)months and a l-year cumulative relapse rate of 13.1％.The overall median value of WT1 before transplantation,3 months after transplantation,and 6 months after transplantation was 0.26％(range:0％-23.64％),with an upper quartile value of 0.74％.No statistically significant differences in WT1 expression levels were observed among the pre-transplantation,3-month post-transplantation,and 6-month post-transplantation time points(P=0.227).Univariate analysis showed that patients with WT1 levels＞0.74％at 3 months post-transplantation had a higher 1-year relapse rate(P=0.029)and lower 3-year OS rate(P＜0.001)compared to patients with WT1 levels ≤0.74％.Other significant factors affecting 1-year relapse included stem cell source(P=0.041)and chronic graft-versus-host disease(cGVHD)(P=0.013).For 3-year OS,additional influencing factors were genetic high risk(P=0.048)and stem cell source(P=0.016).Multivariate analysis revealed that WT1 level＞0.74％at 3 months post-transplantation had a trend to affect 1-year relapse rate(HR=3.309,95％CI:0.958-11.431,P=0.058),while the absence of cGVHD was an independent risk factor for 1-year relapse(HR=3.473,95％CI:0.749-16.100,P=0.037).Only WT1 level＞0.74％at 3 months post-transplantation was an independent risk factor for 3-year OS(HR=6.886,95％CI:2.402-19.738,P＜0.001).Conclusion:High WT1 expression level at 3 months post-transplantation in AML patients undergoing allo-HSCT during CR1 affects the 1-year relapse rate and 3-year OS,and is an independent risk factor affecting 3-year OS.These findings suggest that dynamic monitoring of WT1 expression levels has certain value in prognostic assessment of AML patients who received allo-HSCT during CR1.

Objective To investigate the correlation between seminal plasma oxidation-reduction potential(ORP)and normalized ORP(nORP)with sperm DNA fragmentation index(DFI),high DNA staining(HDS),and motility parameters,and to explore whether these parameters can be used as biomarkers for evaluating male fertility potential.Methods A total of 309 men who attended the andrology clinic at the Department of Reproductive Medicine,General Hospital of Northern Theater Command,between October 2023 and December 2023,were selected in this study.We analyzed the correlation between seminal plasma ORP and nORP and parameters such as DFI using the electrode method,HDS,age,days of abstinence,semen volume,total sperm count,sperm concentration,sperm motility(forward motile sperm count,forward motile sperm percentage,non-forward sperm count,non-forward sperm percentage,immotile sperm count,immotile sperm percentage,and sperm motility rate),and motility indices(curvilinear velocity[VCL],straight-line velocity[VSL],average velocity[VAP],linear index[LIN],straightness index[STR],and wobble coefficient[WOB]).The nORP results were divided into three groups and analyzed in relation to each of the parameters mentioned above.A generalized linear regression model was used to assess the relationship between nORP and HDS,with smooth curve fitting performed to visualize this relationship.Results Pearson and Spearman correlation analyses revealed that both ORP and nORP were positively correlated with semen volume,percentage of immotile sperm,and DFI,but negatively correlated with forward motile sperm count,percentage of forward motile sperm,sperm motility rate,VSL,and VAP(P＜0.05).Additionally,ORP was positively correlated with abstinence days and immotile sperm count(P＜0.05)and negatively correlated with LIN,STR,and WOB(P＜0.05).However,nORP was positively correlated with HDS(P＜0.05),and negatively correlated with total sperm count,sperm concentration,non-forward sperm count,non-forward sperm percentage,immotile sperm count,and VCL(P＜0.05).The nORP group showed significant differences in semen volume,total sperm count,sperm concentration,forward motile sperm count,forward motile sperm percentage,non-forward sperm count,non-forward sperm percentage,immotile sperm count,immotile sperm percentage,sperm motility rate,VCL,VSL,VAP,DFI,and HDS(P＜0.05).A smooth curve fitting of nORP and HDS,adjusted for age and abstinence days,demonstrated a linear upward trend in nORP as HDS increased.Conclusion Both ORP and nORP are associated with sperm DFI and key motility parameters.Moreover,nORP is closely related to HDS,which reflects sperm maturity,offering a more comprehensive indicator of semen quality.Thus,nORP can be utilized as a biomarker for assessing male fertility potential.

Objective:To investigate the serum level of soluble vascular cell adhesion molecule-1 (sVCAM-1) in patients with multiple myeloma (MM) and its effect on the therapeutic effect of daletumab.Methods:A total of 126 MM patients admitted to the Affiliated Hospital of Jining Medical College from June 2019 to June 2021 were retrospectively selected as the observation group, and 120 healthy subjects in the same period were selected as the control group. The observation group was treated with daletumab. The level of sVCAM-1 in the observation group and the control group was compared, and the relationship between serum sVCAM-1 level and clinicopathological features, different Durie-Salmon (DS) stages, different International Staging System(ISS) stages and treatment outcome of MM patients were analyzed.Results:The serum level of sVCAM-1 in the observation group was higher than that in the control group: (797.69 ± 119.73) μg/L vs. (210.55 ± 73.77) μg/L, there was statistical difference ( P<0.01). There were no statistical differences in serum sVCAM-1 level among MM patients with different sex, body mass index, blood calcium, serum albumin, hemoglobin, lactate dehydrogenase and diagnostic type ( P>0.05). The serum level of sVCAM-1 in MM patients DS stage Ⅰ, Ⅱ and Ⅲ were (649.29 ± 101.02), (694.36 ± 109.88) and (729.66 ± 120.44) μg/L, there was statistical difference ( F = 5.12, P<0.01). The serum level of sVCAM-1 in MM patients with ISS stage Ⅰ, Ⅱ and Ⅲ were (648.73 ± 99.77), (701.05 ± 107.83) and (765.82 ± 111.07) μg/L, there was statistical difference ( F = 11.46, P<0.01). After treatment, the serum level of sVCAM-1 in MM patients with complete remission, partial remission and relapse were (234.05 ± 90.73), (445.36 ± 97.11) and (793.05 ± 121.03) μg/L, there was statistical difference ( F = 245.15, P<0.01). The results of receiver operating characteristic (ROC) curve analysis showed that when the cut-off value of serum sVCAM-1 level was 58.50 μg/L, the area under the curve (AUC) was 0.762 (95% CI 0.699 - 0.825, P<0.01), the sensitivity was 80.95%, the specificity was 67.50%, and the accuracy was 74.39%. Conclusions:The level of serum sVCAM-1 in MM patients is significantly increased, and the higher the level of SVCAM-1, the worse the prognosis of patients, which can be used as one of the indicator to predict the therapeutic effect of MM patients.

Totally 45 in vitro diagnostic(IVD)reagent manufacturing enterprises were investigated in terms of the use and quality control of nitrocellulose(NC)membranes.The influences of the NC membrane performance indexes on IVD reagent quality were analyzed,including the within-run stability,crawling speed,protein absorption and pore size.The defficiencies of the enterprises were pointed out in the acceptance standard of NC membrane,and some suggestions were put forward including enhancing the performance of domestic NC membrane,improving the acceptance standard,optimizing NC membrane supply chain management by the enterprises and strengthening the internal management of the enterprises.References were provided for the development of IVD industries.[Chinese Medical Equipment Journal,2025,46(7):75-80]

Objective:To construct an indicator system for a nursing science and technology innovation curriculum guided by innovation competence, in order to provide a reference for cultivating innovation ability in nursing students.Methods:The overall research period was from March to December 2024. A nursing innovation curriculum indicator framework was initially developed through literature analysis and brainstorming. From October to December 2024, 19 experts from nine hospitals or universities across five provinces and cities were selected via purposive sampling to participate in two rounds of Delphi consultation. Revisions were made based on expert feedback.Results:Both rounds of expert consultation achieved a 100% response rate. The authority coefficient of the experts was 0.92. The final indicator system included four curriculum elements: course content, course objectives, teaching methods, and assessment, encompassing 14 first-level indicators and 40 second-level indicators.Conclusions:The innovation-oriented indicator system for nursing science and technology education demonstrates good scientific validity and reliability. It offers a foundational framework for advancing innovation-focused nursing education and curriculum design.

Objective To propose a preventive maintenance strategy of the heating,ventilation and air-conditioning(HVAC)system in pharmacy intravenous admixture services(PIVAS)based on the reliability centered maintenance(RCM)method so as to provide references for PIVAS equipment maintenance.Methods Firstly,a HVAC system RCM review team was formed,and the failure modes and impacts of important functional components of the equipment were analyzed to clarify the consequences of the failure of each functional component under the premise of ensuring the safety and integrity of the equipment and with the goal of minimizing the loss of maintenance downtime and the consumption of maintenance resources.Secondly,with a standardized logical decision-making procedure the preventive maintenance strategy was determined and implemented based on the consequences of functional failure.Finally,statistical analyses were carried out on such equipment indicators as performance parameter qualification rate,failure rate and maintenance cost before and after the RCM method-based strategy was executed,in order to evaluate the efficacy of the strategy.Results The RCM method-based preventive maintenance strategy had the performance qualification rate increased from 97.47%to 99.06%(χ2=24.139,P<0.01),the failure rate decreased from 0.24%to 0.03%(χ2=13.519,P<0.01)and the maintenance cost reduced by 11.5%,from RMB 134,200 to 118,700.Conclusion The RCM method-based preventive maintenance strategy provides reliable equipment for PIVAS and lowers the maintenance cost effectively,and references are given for the development of automated and intelligent equipment maintenance strategies for PIVAS.[Chinese Medical Equipment Journal,2025,46(10):78-83]