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.

Ultrasonic bone-cutting instruments have gained traction in open and minimally invasive spine surgery because of their selective bone-cutting capability while preserving adjacent soft-tissue structures. However, their use in uniportal endoscopic spine surgery has not been reported in the peer-reviewed literature. This technical note describes a uniportal endoscopic surgical technique that uses an ultrasonic osteotome for uniportal endoscopic lumbar interbody fusion (U-ELIF) and reports preliminary outcomes. Specifically, we describe the technical nuances of ultrasonic osteotome use in U-ELIF and evaluate its feasibility and safety, with an emphasis on minimizing complications.

Objective: To analyze the time-related factors influencing daily blood donation attendance at fixed donation sites in Nanjing and the heterogeneity among sites, and to provide evidence for optimizing site layout and operational management. Methods: Based on daily attendance data from 17 fixed donation sites in Nanjing for the full year of 2025, matched with contemporaneous meteorological and air quality data, a two-stage negative binomial generalized linear mixed model (NB-GLMM) was constructed. In the second stage, site-level static characteristics (number of scenic spots, daily operating hours, number of shopping malls, public transport stops, hospitals, universities, and residential buildings) were additionally incorporated. Model comparison was performed using Akaike information criterion (AIC). Significance of random effects was tested by likelihood ratio test, and site-specific random effects were extracted using empirical Bayes estimation. Robustness of the results was verified through residual diagnostics and multiple sensitivity analyses. Results: The first-stage model showed that rainfall was associated with lower attendance (IRR=0.943, P<0.001); holidays were associated with higher attendance (IRR=1.162, P<0.001); Saturdays and Sundays showed higher attendance than Mondays, with the strongest association on Sundays (IRR=1.195, P<0.001). Temperature exhibited a non-linear relationship (linear term P=0.051, quadratic term P=0.036). The joint results of the linear and quadratic terms for temperature suggested a nonlinear relationship between attendance and temperature. After introducing static characteristics in the second stage, the number of scenic spots (IRR=1.372, P=0.008) and daily operating hours (IRR=1.823, P<0.001) were found to be positively associated with attendance, while the numbers of shopping malls, public transport stops, hospitals, universities, and residential buildings showed no significant effects. Conclusion: Daily blood donation attendance at fixed sites in Nanjing is influenced by multiple factors including weather, day of the week, holidays, and site surroundings. Substantial heterogeneity exists among different sites; extended operating hours and more nearby scenic spots are associated with increased attendance.

Objective To evaluate the performance and clinical application value of EasyNAT nucleic acid test kits for influenza A virus(Flu A),influenza B virus(Flu B),and Mycoplasma pneumoniae(MP)based on cross-primed amplification(CPA)technology.Methods In compliance with relevant standards,the detection limits,cross-reactivity,and anti-interference abilities of the EasyNAT Flu and MP nucleic acid test kits were validated using influenza(Flu)and MP reference materials.Additionally,a total of 811 suspected Flu or MP infection patients who visited the hospital from January 1 to July 5,2024 were enrolled in this study.Their oropharyngeal swabs were collected and tested using reverse transcription quantitative polymerase chain reac-tion(RT-qPCR)as the gold standard to evaluate the concordance and compliance of the EasyNAT Flu and MP nucleic acid test kits,thus assessing their clinical application value.Results The EasyNAT Flu and MP nucleic acid test kits yielded positive results in 8 repeated tests at the detection limit concentrations of the ref-erence materials.No pathogen-specific peaks were detected in Flu and MP negative samples that had been spiked with common cross-reacting pathogens.Furthermore,the detection results of Flu or MP positive sam-ples were unaffected by the presence of interfering substances.Among the 811 clinical samples,the sensitivity of the EasyNAT Flu A test was 97.33%,and the sensitivity for Flu B was 98.47%,both with a specificity of 98.04%.The EasyNAT MP nucleic acid test showed a sensitivity of 97.95%and specificity of 99.03%.The results demonstrated a high degree of agreement with RT-qPCR(Kappa values were over 0.900).Further-more,detection rates of the EasyNAT Flu test kits showed no difference in positive samples with different Ct values(Flu A:χ2=4.20,P=0.08;Flu B:χ2=2.22,P=0.31)and MP test kits showed significant differences in positive samples with different cycle threshold values(χ2=11.84,P<0.01).Conclusion The EasyNAT Flu and MP nucleic acid test kits exhibit excellent detection performance with high sensitivity and specificity.Moreover,the products are easy to operate,offer rapid detection,and provide accurate results.These features make them suitable for primary healthcare settings,with significant potential for clinical application.

Objective To investigate the expressions of annexin A2 and glycogen synthesis kinase-3β(GSK-3β)in cutaneous squamous cell carcinoma(CSCC)tissues,and to analyze their correlation with CSCC as well as their clinical pathological diagnostic value.Methods The pathological tissues of 68 patients with CSCC and 40 patients with keratoacanthoma(KA)who underwent surgical treatment in the Department of Dermatology of the Second Hospital of Nanning from October 2020 to May 2024 were collected,and the surrounding normal skin tissues of 32 patients with benign skin diseases were used as controls.The expressions of annexin A2,GSK-3β and β-catenin were detected by immunohistochemistry and Western blotting.Spearman was used to evaluate the correlation between the expressions of annexin A2 and GSK-3β and the pathological characteristics in CSCC.The receiver operating characteristic(ROC)curve was drawn to analyze the clinical diagnostic value of annexin A2 and GSK-3β in CSCC.Results Compared with the normal skin tissues,the expressions of annexin A2 and β-catenin in CSCC increased,and GSK-3β decreased(P＜0.05);Compared with the KA tissues,the expression of annexin A2 in CSCC tissues increased(P＜0.05).The expression of annexin A2 was negatively correlated with that of GSK-3β in CSCC(r=-0.3901,P＜0.01).GSK-3β expression was related to tissue differentiation,with lower expression in poorly differentiated patients'cancer tissues(P＜0.05).The sensitivity of annexin A2 and GSK-3β for diagnosis of CSCC was 85.3%and 41.2%,respectively,with specificities of 46.9%and 84.4%respectively.The sensitivity of annexin A2 for distinguishing between CSCC and KA was 85.3%,with a specificity of 40.0%.Conclusion Annexin A2 and GSK-3β may be used as potential biomarkers for the early diagnosis or differential diagnosis of CSCC,and play important roles in the development of CSCC.Their mechanism may be related to the activation of Wnt/β-catenin signaling pathway.

Objective To explore the value of MRI radiomics based on the frontal and temporal lobes combined with multiple machine learning algorithms in the diagnosis of mild cognitive impairment(MCI).Methods Patients who underwent cranial MR examination were retrospectively selected.According to the inclusion and exclusion criteria,a total of 173 subjects were finally included and randomly divided into training set and test set in a ratio of 7∶3.After delineating the regions of interest(ROI)of the frontal and temporal lobes on T2-fluid attenuated inversion recovery(FLAIR)images,radiomics features were extracted based on the Pyradiomics data package.Features were screened through inter-and intraclass correlation coefficient(ICC),independent samples t-test,and the LightGBM algorithm.Diagnostic models were constructed using support vector machine(SVM),random forest(RF),decision tree(DT),K-nearest neighbor(KNN),gradient boosting decision tree(GBDT),and extreme gradient boosting(XGBoost)combined with 10-fold cross-validation respectively.The training set was further divided into 9 training data sets and 1 validation data set through 10-fold cross-validation,and the hyperparameters were optimized through iterative cycles.The diagnostic efficacy of the model was evaluated by receiver operating characteristic(ROC)curve and area under the curve(AUC),and the DeLong test was applied to compare the differences between different models.Results The AUC of the radiomics models constructed by SVM,DT,RF,KNN,GBDT,XGBoost in the training set were 0.951,0.992,0.998,0.957,1.000,and 1.000 respectively,in the validation set were 0.890,0.843,0.934,0.878,0.930,and 0.945 respectively,and in the test set were 0.902,0.711,0.899,0.849,0.889,and 0.882 respectively.Conclusion MRI radiomics based on the frontal and temporal lobes combined with multiple machine learning algorithms can diagnose MCI,and the model constructed based on SVM shows the highest diagnostic value.

Objective:To investigate the early clinical efficacy of arthroscopic suture half-tie combined with Versalok anchor fixation in the treatment of avulsion fracture of the tibial insertion of anterior cruciate ligament (ACL) in adults.Methods:A retrospective study was conducted to analyze the clinical data of the 26 adult patients with avulsion fracture of the ACL tibial insertion who had been treated by arthroscopic suture half-tie combined with Versalok anchor fixation between June 2020 and December 2022 at Sports Medicine Center, Honghui Hospital, Xi'an Jiaotong University. This cohort consisted of 12 males and 14 females with an age of (26.6±7.5) years. According to the Meyers-McKeever classification, there were 9 cases of type Ⅱ, 12 cases of type Ⅲ, and 3 cases of type Ⅳ. The time from injury to surgery was (9.3±6.8) days. Follow-up evaluations were conducted at postoperative 3 days, 3 months, and 12 months, including imaging assessments for fracture healing, clinical examinations of knee flexion and extension, anterior drawer and Lachman tests to assess the ACL tension. At the last follow-up, KT-2000 test was performed to quantify ACL laxity, and visual analogue scale (VAS) and Lysholm knee score were used to assess postoperative pain and functional recovery.Results:Primary wound healing was achieved in all the 26 patients. One patient required ACL reconstruction due to laxity and mild extension limitation at postoperative 6 months. The other 25 patients were followed for (15.4±3.2) months. At postoperative 12 months, X-rays showed good bone healing at the fracture site of the tibial intercondylar eminence, and both the Lachman and anterior drawer tests were negative. At postoperative 3 months and the last follow-up, the active knee extension and flexion were 128.8° (121.2°, 138.3°) and 52.7° (38.2°, 63.2°), respectively, significantly better than the preoperative value [52.7° (38.2°, 63.2°)] ( P<0.05), and the KT-2000 test showed that the disparity between the affected and healthy sides were 2.5 (1.2, 4.2) mm and 1.2 (0.8, 3.4) mm, respectively, significantly smaller than the preoperative value [7.1 (5.2, 11.2) mm] ( P<0.05). At postoperative 3 and 12 months, the VAS pain scores were significantly decreased and the knee Lysholm scores were significantly improved compared with the preoperative values ( P<0.05). Conclusion:In the treatment of avulsion fracture of the ACL tibial insertion in adults, arthroscopic suture half-tie combined with Versalok anchor fixation is a minimally invasive, reliable, and straightforward technique, leading to excellent early clinical outcomes.

Objective To explore the distribution patterns of traditional Chinese medicine(TCM)syndromes in patients with metabolic syndrome caused by antipsychotic drugs.Methods A standardized TCM syndrome survey was performed to collect diagnostic information from 160 patients diagnosed with metabolic syndrome due to antipsychotic drug use.Subsequent frequency analysis,cluster analysis,and Bayesian network analysis were carried out.The syndrome pattern distribution was ultimately determined through relevant literatures and expert opinions.Results Five TCM syndromes were identified through frequency,cluster,and Bayesian network analyses.The most common syndrome was qi deficiency with phlegm-dampness(30％),followed by spleen deficiency with phlegm-Heat(23.75％),qi and yin deficiency Pattern(21.88％),yin deficiency with damp-heat(17.50％),and stomach fire hyperactivity pattern(6.88％).Conclusion The pathogenesis of antipsychotics-induced metabolic syndrome involves a complex interplay of deficiency and excess factors.The primary disease is mainly located at the spleen and stomach,with involvement of the liver,kidney,and heart.Pathogenic factors include qi deficiency,yin deficiency,dampness,heat,pathogenic fire,and phlegm.

Objective·To study the effect of neodymium-doped yttrium aluminum perovskite(Nd:YAP)laser biostimulation on the proliferation and osteogenic differentiation of human periodontal ligament cells(hPDLCs)and its possible mechanism.Methods·Five premolars removed for orthodontic reasons were collected from Changzhou Stomatological Hospital,and the periodontal ligament tissues from the middle 1/3 of the roots were taken to culture hPDLCs in vitro.The cells were irradiated with the biostimulation function[G(-)mode]of the Nd:YAP laser.According to the irradiation time,the cells were divided into a control group(without laser irradiation),and groups irradiated for 5 s,10 s,15 s,20 s and 30 s.The cell counting kit-8(CCK-8)method was used to detect the proliferation of hPDLCs in each group.After osteogenic differentiation was induced,the alkaline phosphatase(ALP)content and activity level of the cells were detected using an ALP staining kit and an ALP activity detection kit.The calcium salt level of the cells was evaluated by alizarin red S staining and calcium quantitative analysis.The expression of genes and proteins related to the WNT/β-catenin signaling pathway,including dickkopf-related protein 1(DKK-1),β-catenin,and runt-related transcription factor 2(RUNX2),was analyzed by quantitative real-time PCR(qPCR)and Western blotting.Results·The results of CCK-8 showed that the proliferation level of cells in the 10 s,15 s,20 s,and 30 s groups was enhanced from 3 d after irradiation(all P<0.05).After induction of osteogenic differentiation,ALP content,activity,and calcium salt level in the laser irradiation groups increased with the extension of irradiation time(all P<0.05).The results of qPCR and Western blotting analysis showed that the expression levels of the DKK-1 gene and protein in the laser irradiation groups decreased with the extension of irradiation time.However,the expression levels of β-catenin and RUNX2 genes and proteins increased significantly with the extension of irradiation time;there were statistically significant differences between the 15 s,20 s,and 30 s groups and the control group(all P<0.05).Conclusion·Nd:YAP laser biostimulation may promote proliferation and osteogenic differentiation of hPDLCs through WNT/β-catenin signaling pathway.

Solid-phase extraction(SPE)combined with surface-enhanced Raman spectroscopy(SERS)has emerged as a promising analytical technique for detection and analysis of trace components in complex sample matrices.SPE enriches analytes through selective adsorption and solvent elution,effectively increasing the concentration and signal intensity.SERS enables ultra-sensitive and highly selective molecular analysis through the use of SERS-active substrates engineered to amplify Raman signals.The integration of these two techniques overcomes the limitations of conventional Raman spectroscopy in low-concentration detection field,while significantly improving sample preparation efficiency and analytical accuracy.This review provided a comprehensive overview of the characteristics of three SPE-SERS coupling modes,including two-step,one-step,and online integration.Special emphasis was placed on recent advancements in one-step SPE-SERS approaches based on novel functional adsorbent materials such as graphene,metal-organic frameworks,covalent organic frameworks,and molecularly imprinted polymers.Furthermore,future directions and development prospects of SPE-SERS technology were discussed.