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.

With the rising incidence of colon cancer, several studies have indicated that aquaporin 1 (AQP1) expression is associated with the development of colon cancer. This study aims to elucidate the potential molecular mechanisms between them. Methods: We screened data from The Cancer Genome Atlas (TCGA) database and retrospectively examined AQP1 protein expression in 127 colon cancer patients to analyze the relationship between AQP1 expression and pathological stages, prognosis. We created stable colon cancer cell lines with differential AQP1 expression, the effect of AQP1 expression on the proliferation and migration of colon cancer cells was assessed by in vitro and in vivo studies, and explored potential molecular mechanisms through Western blotting. Results: High AQP1 expression was associated with poorer survival (overall survival [OS], p = .028) in colon cancer patients from the TCGA database. Similarly, retrospective clinical data indicated that high AQP1 expression was associated with reduced disease-free survival and OS (p = .036 and p = .017, respectively). The low-expressing AQP1 colon cancer cells exhibited a decrease in proliferation and migration ability of colon cancer cells compared to the overexpressing AQP1 group (p < .05) in vitro and in vivo. Immunohistochemistry and western blotting experiments validated heightened expression of N-cadherin, vimentin, and claudin- 1 in the tumor tissues of the overexpressing AQP1 group. Conversely, reduced AQP1 expression resulted in decreased expression of claudin- 1. Conclusions: AQP1 correlates with unfavorable prognosis in colon cancer and potentially enhances the proliferation and migration of colon cancer by up-regulating claudin-1 expression.

Objective To explore the relationship between PANoptosis and hepatic ischemia-reperfusion injury (HIRI), and to screen the key genes of PANoptosis in HIRI. Methods PANoptosis-related differentially expressed genes (PDG) were obtained through the Gene Expression Omnibus database and GeneCards database. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA) were used to explore the biological pathways related to PDG. A protein-protein interaction network was constructed. Key genes were selected, and their diagnostic value was assessed and validated in the HIRI mice. Immune cell infiltration analysis was performed based on the cell-type identification by estimating relative subsets of RNA transcripts. Results A total of 16 PDG were identified. GO analysis showed that PDG were closely related to cellular metabolism. KEGG analysis indicated that PDG were mainly enriched in cellular death pathways such as apoptosis and immune-related signaling pathways such as the tumor necrosis factor signaling pathway. GSEA results showed that key genes were mainly enriched in immune-related signaling pathways such as the mitogen-activated protein kinase (MAPK) signaling pathway. Two key genes, DFFB and TNFSF10, were identified with high accuracy in diagnosing HIRI, with areas under the curve of 0.964 and 1.000, respectively. Immune infiltration analysis showed that the control group had more infiltration of resting natural killer cells, M2 macrophages, etc., while the HIRI group had more infiltration of M0 macrophages, neutrophils, and naive B cells. Real-time quantitative polymerase chain reaction results showed that compared with the Sham group, the relative expression of DFFB messenger RNA in liver tissue of HIRI group mice increased, and the relative expression of TNFSF10 messenger RNA decreased. Cibersort analysis showed that the infiltration abundance of naive B cells was positively correlated with DFFB expression (r=0.70, P=0.035), and the infiltration abundance of M2 macrophages was positively correlated with TNFSF10 expression (r=0.68, P=0.045). Conclusions PANoptosis-related genes DFFB and TNFSF10 may be potential biomarkers and therapeutic targets for HIRI.

Objective To explore the regulatory mechanism of NFE2L2/KEAP1 in alveolar bone repair induced by periodontitis.Methods A rat periodontitis model was established and divided into four groups:Control group(n=6);Periodontitis model group(n=6);Periodontitis+lentivirus empty vector group(n=6);Periodontitis+NFE2L2 overexpression plasmid group(n=6).Histopathological changes in each group were observed using HE staining.TRAP staining was used to detect osteoclast positivity,while ELISA was employed to measure inflammatory cytokine levels in tissues.Immunofluorescence and qPCR were used to detect NFE2L2 expression,and western blot was used to assess the expression of osteogenic proteins ALPL2,RUNX2,and COL1.Primary periodontal ligament cells(hPDLCs)were cultured,and cells were transfected to overexpress NFE2L2 and KEAP1.The cells were divided into six groups:Normal group;Model group;pcDNA-NC group;pcDNA-NFE2L2 group;pc-NFE2L2+pcDNA-NC group;pc-NFE2L2+pcDNA-KEAP1 group.A cellular model was established,and the morphology of primary hPDLCs was observed under a microscope.Cell proliferation was assessed using CCK-8.Osteogenic mineralization was observed using alizarin red staining,and western blot was used to detect osteogenic proteins and autophagy markers.Cell migration was observed using a scratch assay.Results(1)After model induction,redness,swelling of the gums,extensive inflammatory infiltration,and alveolar bone resorption were observed,confirming successful model establishment.Partial tissue recovery occurred after NFE2L2 overexpression via lentivirus.(2)After model induction,osteoclast positivity increased,confirming successful model establishment.Overexpression of NFE2L2 reduced osteoclast positivity(P<0.001).(3)After model induction,levels of IL-1β,IL-10,and TNF-α were significantly higher than in the normal group(P<0.05),confirming successful model establishment.Transfection with NFE2L2 lentivirus reduced inflammatory cytokine levels(P<0.0001).After model induction,osteogenic protein expression decreased compared to the normal group,but overexpression of NFE2L2 increased osteogenic protein expression(P<0.05).(5)LPS treatment significantly reduced cell viability,while NFE2L2 overexpression enhanced it(P<0.0001).(6)LPS treatment reduced calcified nodules,while NFE2L2 overexpression increased them.Addition of pcDNA-KEAP1 reduced mineralized nodules.(7)LPS treatment decreased osteogenic protein expression,while NFE2L2 overexpression increased it.However,addition of pcDNA-KEAP1 reduced osteogenic protein expression(P<0.05).(8)LPS treatment reduced cell migration,whereas NFE2L2 overexpression enhanced it(P<0.0001).(9)Expression of autophagy markers decreased after LPS treatment,but increased after transfection with NFE2L2 plasmid.However,addition of pcDNA-KEAP1 reduced the expression of autophagy markers(P<0.05).Conclusion This study identified the regulatory role of NFE2L2/KEAP1 in periodontitis,providing a scientific basis for the treatment of periodontitis.

Objective To conduct preoperative simulations of three different osteotomy line design schemes under centric occlusion based on two distinct material assignment methods,evaluate biomechanical properties of the models,and explore which osteotomy line design schemes are more suitable for different types of mandibles.Methods Three types of mandibles were selected,and CT images were obtained for three-dimensional(3D)reconstruction.Material assignment was completed using the cortical/cancellous bone assignment method and the gray value assignment method.Osteotomy was simulated according to the three osteotomy line design schemes,followed by finite element analysis.Results In all simulation results of the mandibles,the maximum stress was 81.10 MPa,the maximum strain was 0.035 52,and the maximum displacement was 432.4 μm.The stress distributions obtained by the cortical/cancellous bone assignment method showed a larger stress distribution range than that that by the gray value assignment methods,but the maximum stress,strain,and displacement were generally lower.For the outflare type and common type mandibles,Scheme 1 showed lower maximum stress,strain,and displacement under both material assignment methods,but no clearly suitable scheme was found for the retracted type.Conclusions The outflare type and common type mandibles are more suitable for adopting the osteotomy line design scheme of Scheme 1.For the retracted type,other mandibular angle osteotomy plastic surgery methods may be considered to ensure better biomechanical characteristics.Whether choosing the osteotomy line design scheme or the modeling material assignment method,it is necessary to make the final decision based on the specific analysis objective and resource conditions.

Subarachnoid hemorrhage (SAH) is a serious intracranial hemorrhage characterized by acute bleeding into the subarachnoid space. The effects of shikonin, a natural compound from the roots of Lithospermum erythrorhizon, on oxidative stress and blood–brain barrier (BBB) injury in SAH was evaluated in this study. A rat model of SAH was established by endovascular perforation to mimic the rupture of intracranial aneurysms. Rats were then administered 25 mg/kg of shikonin or dimethylsulfoxide after surgery. Brain edema, SAH grade, and neurobehavioral scores were measured after 24 h of SAH to evaluate neurological impairment. Concentrations of the oxidative stress markers superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA) in the brain cortex were determined using the corresponding commercially available assay kits. Evans blue staining was used to determine BBB permeability. Western blotting was used to quantify protein levels of tight junction proteins zonula occludens-1, Occludin, and Claudin-5. After modeling, the brain water content increased significantly whereas the neurobehavioral scores of rats with SAH decreased prominently. MDA levels increased and the levels of the antioxidant enzymes GSH and SOD decreased after SAH. These changes were reversed after shikonin administration. Shikonin treatment also inhibited Evans blue extravasation after SAH. Furthermore, reduction in the levels of tight junction proteins after SAH modeling was rescued after shikonin treatment. In conclusion, shikonin exerts a neuroprotective effect after SAH by mitigating BBB injury and inhibiting oxidative stress in the cerebral cortex.

BACKGROUND:Patch clamp technique has been developed for more than 40 years as the"gold standard"for the study of ion channels.However,the research content of scientific research institutions is relatively independent,and the existing research results are not systematically summarized,which leads to the phenomenon of high repeatability and weak innovation in the existing research.Therefore,it is urgent to make a comprehensive review of patch clamp technology to clarify the current research status,hot spots,and future development direction.OBJECTIVE:To summarize the research status and development trend of patch clamp technique in recent 10 years.METHODS:Publications on patch clamp technology from 2013 to 2023 were collected using the Web of Science core collection database.CiteSpace and VOSviewer software were used to quantify the number of publications and analyze the network of literature entries,including countries,institutions,journals,authors,keywords,highly cited literature,and co-cited references.RESULTS AND CONCLUSION:(1)In recent 10 years,the research in the field of patch clamp technology has gradually entered a stage of stable development.(2)China and the United States are the leading countries in this regard.The Chinese Academy of Sciences is an institution with core influence.Journal of Neuroscience is the main publication.Park,Won Sun team(Jeonbuk National University)and Chu,Li team(Hebei Key Laboratory of Chinese Medicine Research on Cardio-Cerebrovascular Disease)have made outstanding contributions in this field,but there is less collaboration and communication between the teams and no network cooperation model has been formed.(3)Patch clamp technology is mainly used in the electrophysiological characteristics of the nervous system and the pathological mechanism of the disease,which is the focus of researchers'continuous attention.(4)In the study of electrophysiological characteristics of cardiovascular system and its pathological mechanism,the electrophysiological characteristics of primary cardiomyocytes and induced pluripotent stem cell-derived cardiomyocytes and the pathological mechanism of atrial fibrillation,cardiotoxicity,sudden cardiac death,hypertension and other cardiovascular diseases have been the focus of research in recent years.(5)In the application of patch clamp technology combined with other biotechnology,it will be an important research direction to focus on the cross fusion with optogenetics,two-photon calcium imaging and other technologies.(6)In the research of drug screening and identification of therapeutic targets,especially the research of patch clamp technology and traditional Chinese medicine compound,it will become a great help in the future research of component traditional Chinese medicine.

Objective:To investigate the efficacy of using a single Kocher-Langenbeck approach to improve anterior column screw fixation for the treatment of transverse + posterior wall acetabular fractures.Method:A retrospective analysis was conducted of the data of 21 patients who had been treated with a single Kocher-Langenbeck approach to improve anterior column screw fixation for transverse + posterior wall acetabular fractures at Department of Orthopaedics, Weifang People's Hospital from May 2021 to May 2023. There were 11 males and 10 females with an age of (51.3±15.5) years. The time from injury to surgery was (10.4±3.6) days. The anterior column screw fixation was improved as follows: screws were implanted 1 to 2 cm in front of the apex of the great notch of the external iliac plate of the pelvis to penetrate out at the upper branch root of the pubic ramus, with the whole screw fully located within the bony channel. Such indexes were recorded as surgical time, intraoperative blood loss, intraoperative fluoroscopy frequency for improved placement of anterior column screws, fracture healing time, incidence of complications, and fracture reduction and hip joint function at the last follow-up.Results:For this cohort, surgical time was (120.9±40.7) minutes, intraoperative blood loss (330.2±65.9) mL, and intraoperative fluoroscopy frequency for improved placement of anterior column screws (6.2±2.3) times. The 21 patients were followed up for (21.5±5.9) months. All fractures achieved bony union after (3.2±1.1) months. Incision fat liquefaction occurred in 1 patient after surgery and symptoms of sciatic nerve injury were observed in 2 patients. Follow-ups observed no such complications as loosening or breakage of internal fixators. According to the Matta scoring criteria, the quality of fracture reduction at the last follow-up was evaluated as anatomical reduction in 15 cases, as satisfactory reduction in 4 cases, as acceptable reduction in 2 cases, giving a satisfaction rate of 90.5% (19/21). According to the improved Merle d'Aubigné & Poster scoring criteria, hip function was evaluated as excellent in 16 cases, as good in 2 cases, and as fair in 3 cases, giving an excellent and good rate of 85.7% (18/21).Conclusion:It is effective to use a single Kocher-Langenbeck approach to improve anterior column screw fixation for the treatment of transverse + posterior wall acetabular fractures, showing advantages of reduced surgical time and intraoperative bleeding.

Aim To explore the relationship between serum levels of proprotein convertase subtilisin/kexin type 9(PCSK9),macrophage migration inhibitory factor(MIF)and the severity of coronary artery lesions in patients with coro-nary heart disease(CHD).Methods A cross-sectional study was conducted involving 139 patients with CHD and 69 control subjects who underwent coronary angiography during the same period,all of whom were admitted to the People's Hospital of Xinjiang Uygur Autonomous Region from November 2023 to May 2024.Clinical data and coronary angiography results were collected,and the severity of coronary artery stenosis was quantitatively assessed using the Gensini score.Pa-tients with the Gensini scores＞0 were classified into three groups based on tertiles:the mild stenosis group(1～18 points,54 cases),the moderate stenosis group(19～36 points,54 cases),and the severe stenosis group(＞36 points,54 ca-ses).Serum levels of PCSK9 and MIF were measured by ELISA kit.Results Serum levels of PCSK9 and MIF were significantly higher in the CHD group than those in the control group(P＜0.05).Multivariable Logistic regression analy-sis revealed that high levels of serum PCSK9 and MIF were independent risk factors for CHD.Spearman correlation analy-sis showed that serum PCSK9 and MIF levels were positively correlated with Gensini score(rs=0.619 6 and r,=0.411 4,both P＜0.001).Further subgroup analysis showed that serum total cholesterol and low density lipoprotein cholesterol lev-els were significantly increased in patients with high-level PCSK9,while patients with high-level MIF had higher inflamma-tory coefficients such as systemic inflammatory response index(SIRI)and systemic immune-inflammation index(SII)(all P＜0.05).Conclusion Serum levels of PCSK9 and MIF are positively correlated with the severity of coronary artery stenosis.High levels of serum PCSK9 and MIF are independent risk factors for CHD.

Objective:To investigate the predictive value of machine learning-based CT radiomics model for radioresistance in patients with esophageal squamous cell carcinoma (ESCC).Methods:Clinical data of 185 patients with ESCC treated with radical radiotherapy in the First Affiliated Hospital of Anhui Medical University from December 2015 to July 2022 were retrospectively analyzed, and all patients were randomly divided into a training set ( n=129) and a validation set ( n=56) at a ratio of 7 : 3. The radiomics parameters of the primary lesion of esophageal cancer and the surrounding 5 cm region in the patients' CT arterial phase images were extracted, and 6 machine learning methods were used to screen the optimal radiomics model to obtain the optimal radiomics score (Radscore). Independent prognostic predictors of radioresistance in ESCC were obtained by univariate and multivariate logistic regression analyses, which was used as the basis for constructing the nomogram. The predictive performance of different models was compared by the area under the receiver operating characteristic (ROC) curve (AUC). The predictive efficacy and clinical value of the combined model were evaluated using calibration curve, decision curve analysis and clinical impact curve, net reclassification improvement (NRI) and integrated discrimination improvement (IDI). Results:The combined intratumoral and peritumoral radiomics model based on naive Bayesian classifier yielded the optimal prediction performance, with AUC of 0.859 and 0.936 in the training set and validation set, respectively. Multivariate logistic regression analysis showed that Radscore and T stage were the independent prognostic predictors of radioresistance in ESCC patients, and the AUC of the combined model constructed based on these predictors in the training and validation sets were 0.942 and 0.959, respectively. Calibration curve, decision curve analysis and clinical impact curve, net reclassification improvement (NRI) and integrated discrimination improvement (IDI) all indicated higher clinical benefit and more consistent predictive efficacy of the combined model.Conclusions:Machine learning-based CT radiomics model is useful for the prediction of radioresistance in ESCC. The nomogram of radiomics and clinical parameters can further improve the prediction accuracy and provide novel reference for individualized treatment of patients with ESCC.