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.

Background: Murraya, a genus of shrubs and trees in the Rutaceae family, consists of approximately 9 species in China with significant medicinal and horticultural value. However, the phylogeny and taxonomy of Murraya species remain controversial, particularly with respect to Murraya exotica and M. paniculata. Objective: This study aimed to provide insights into the taxonomy, phylogeny, and identification of Murraya. Methods: In this study, the chloroplast (CP) genomes of 7 Murraya species were sequenced, assembled, and subjected to comparative and phylogenetic analyses. Results: The CP genomes of Murraya ranged from 158,573 to 160,817 bp in length and encoded 112 unique genes, including 78 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. Similar to other angiosperms, the inverted repeat regions of the CP genomes exhibited lower sequence divergence than the single-copy regions, and coding regions were more conserved than noncoding regions. Comparative analysis identified several highly variable regions (eg, matK, ycf1, ndhI-ndhA, trnH-GUG-psbA, rpl32-trnL) that could serve as molecular markers for species identification in Murraya. Among these, the ycf1 gene was validated as a useful marker for distinguishing M. exotica from M. paniculata. Positive selection was detected in 10 genes, including rbcL, psaJ, ndhD, ndhF, rpl2, rpl20, ycf1, accD, ccsA, and rpl32. Phylogenetic analysis based on CP genomes supported the recognition of M. exotica and M. paniculata as independent species. Moreover, the phylogenetic trees indicated that Murraya is not monophyletic, with sect. Bergera showing a closer relationship to Clausena. Molecular dating results suggested that the diversification of M. paniculata, M. alata, and M. exotica occurred approximately 9.11 Mya (95% highest posterior density: 4.90-13.87 Mya). Conclusion: These findings provide valuable CP genome data for clarifying the phylogenetic relationships between M. exotica and M. paniculata, and for advancing the study of DNA markers and the evolutionary history of Murraya.

Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

With the widespread adoption of low-dose CT screening and the extensive application of high-resolution CT, the detection rate of sub-centimeter lung nodules has significantly increased. How to scientifically manage these nodules while avoiding overtreatment and diagnostic delays has become an important clinical issue. Among them, lung nodules with a consolidation tumor ratio less than 0.25, dominated by ground-glass shadows, are particularly worthy of attention. The therapeutic challenge for this group is how to achieve precise and complete resection of nodules during surgery while maximizing the preservation of the patient's lung function. The "watershed topography map" is a new technology based on big data and artificial intelligence algorithms. This method uses Dicom data from conventional dose CT scans, combined with microscopic (22-24 levels) capillary network anatomical watershed features, to generate high-precision simulated natural segmentation planes of lung sub-segments through specific textures and forms. This technology forms fluorescent watershed boundaries on the lung surface, which highly fit the actual lung anatomical structure. By analyzing the adjacent relationship between the nodule and the watershed boundary, real-time, visually accurate positioning of the nodule can be achieved. This innovative technology provides a new solution for the intraoperative positioning and resection of lung nodules. This consensus was led by four major domestic societies, jointly with expert teams in related fields, oriented to clinical practical needs, referring to domestic and foreign guidelines and consensus, and finally formed after multiple rounds of consultation, discussion, and voting. The main content covers the theoretical basis of the "watershed topography map" technology, indications, operation procedures, surgical planning details, and postoperative evaluation standards, aiming to provide scientific guidance and exploration directions for clinical peers who are currently or plan to carry out lung nodule resection using the fluorescent microscope watershed analysis method.

ObjectiveTo investigate the effect and mechanism of polysaccharide in water extract of Cyclocarya paliurus （CPWP） in inhibiting benign prostatic hyperplasia （BPH）. MethodsCPWP was obtained by heating reflux， aqueous extraction， alcohol precipitation， and freeze drying. The chemical composition and structural properties of CPWP were analyzed by high performance liquid chromatography with 1-pheny-3-methyl-5-pyrazolone pre-column derivatization and infrared spectroscopy. Male SD rats were randomly assigned into control， model， finasteride （ig 5 mg·kg^-1）， and low-， medium-， and high-dose （ig 50， 75， 100 mg·kg^-1） CPWP groups， with 8 rats in each group. The BPH model was established by subcutaneously injecting propionate testosterone in castrated rats. The rats in the drug intervention groups were administrated with corresponding drugs， and those in the control group were administrated with an equal volume of normal saline each day. After 30 consecutive days， the rats were sacrificed， and the prostate tissue was separated and weighed. The effects of drug interventions on the body weight， prostate wet weight， and prostate index of rats were examined. The prostate tissue was stained with hematoxylin-eosin （HE） for observation of pathological changes. Enzyme-linked immunosorbent assay was employed to measure the level of dihydrotestosterone （DHT）， and immunohistochemical staining was used to detect the expression of steroid 5 alpha-reductase 2 （SRD5A2） and Ki67 in the prostate tissue. ResultsCPWP was identified as a saccharide， with characteristic absorption peaks of saccharides. CPWP showed the total sugar content of 44.15% and molecular weight within the range of 5.5-78.8 kDa， being composed of mannose， rhamnose， galacturonic acid， glucose， galactose， xylose， and arabinose. Compared with the control group， the model group had significantly increased prostate wet weight and prostate index （P<0.01）， thick and tall prostate epithelial cells， increased internal wrinkles， papillary expansion into the cavity， an elevation in DHT level in the serum， and up-regulated expression of SRD5A2 and Ki67 in the prostate tissue （P<0.05， P<0.01）. Compared with the model group， both the finasteride and CPWP groups showed decreases in prostate wet weight and prostate index （P<0.05， P<0.01）， thinned prostate epithelial cells， with only a small portion of internal wrinkles and papillary expansion into the cavity， shortened papillary protrusions， lowered DHT level in the serum， and down-regulated expression of SRD5A2 and Ki67 in the prostate tissue （P<0.01）. Moreover， CPWP exerted effects in a dose-dependent manner. ConclusionCPWP inhibits BPH by regulating the expression of SRD5A2.

The alternative pathway (AP) of the complement system is a key contributor to the pathogenesis of several diseases including paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), C3 glomerular disease (C3G) and age-related macular degeneration (AMD). Complement factor B (CFB) is a trypsin-like serine protein that circulates in the human bloodstream in a latent form. As a key node of the alternative pathway, it is an important target for the treatment of diseases mediated by the complement system. With the successful launch of iptacopan, the CFB small molecule inhibitors has become a current research hotspot, a number of domestic and foreign pharmaceutical companies are actively developing CFB small molecule inhibitors. In this paper, the research progress of CFB small molecule inhibitors in recent years is systematically summarized, the representative compounds and their activities are introduced according to structural types and design ideas, so as to provide reference and ideas for the subsequent research on CFB small molecule inhibitors.

BACKGROUND:β-Defensin has the ability against influenza A virus and inhibits a series of inflammatory responses induced by influenza A virus infection within cells.There have been no reports on whether hemagglutinin 1 from influenza A virus can induce the secretion of mouse β-defensin and interferon-γ when acting in mouse tracheal epithelial cells.OBJECTIVE:To investigate the effect of recombinant hemagglutinin 1 on the production levels of mouse β-defensin and interferon-γ in mouse tracheal epithelial cells.METHODS:Primary mouse tracheal epithelial cell were divided into six groups:blank control group(Control),recombinant hemagglutinin 1 group(200 ng/mL),recombinant hemagglutinin 1+influenza A virus group,influenza A virus group(2×TCID50),recombinant hemagglutinin 1+inactivated influenza A virus group,and inactivated influenza A virus(I)group.After the mouse tracheal epithelial cells in each group were treated for 4,8,or 24 hours,hematoxylin-eosin staining was used for pathological observation.The mRNA levels of mouse β-defensin 2,3,and 4,and interferon-γ were detected by qRT-PCR.The protein levels of mouse β-defensin 2,3,and 4 were measured by enzyme-linked immunosorbent assay,and the protein expression of interferon-γ was detected by western blot.RESULTS AND CONCLUSION:(1)The recombinant hemagglutinin 1 acting alone or in combination with influenza A virus could cause different pathological changes in tracheal epithelial cells.Phenomena such as vacuolation,nuclear pyknosis and cell fusion could be observed in the cells.(2)Compared with the control group,recombinant hemagglutinin 1 alone or in combination with influenza A virus or inactivated influenza A virus significantly induced the production of mouse β-defensin 2,3,and 4(P<0.05)and interferon-γ in mouse tracheal epithelial cells(P<0.05).These results indicate that recombinant hemagglutinin 1 alone or in combination with influenza A virus can induce the production of mouse β-defensin 2,3,and 4 and interferon-γ in mouse tracheal epithelial cells.

Objective·To explore the effect of intermittent theta burst stimulation(iTBS)targeting the left dorsolateral prefrontal cortex(DLPFC)on reducing craving in alcohol-dependent patients during the withdrawal period,as well as its impact on patients'emotions and cognitive functions.Methods·A total of 41 inpatients with alcohol dependence in the withdrawal period were recruited from the Addiction Department of Mental Health Center,Shanghai Jiao Tong University School of Medicine,and randomly assigned to the experimental group(20 patients)and the control group(21 patients).Both groups received routine inpatient treatment for alcohol-dependence.The experimental group received real iTBS stimulation targeting the left DLPFC on the basis of routine inpatient treatment,while the control group received sham stimulation with the same parameters.The intervention course lasted for 2 weeks,with a total of 10 sessions.The Visual Analogue Scale(VAS),Beck Depression Inventory(BDI),and Beck Anxiety Inventory(BAI)were used to measure the craving,anxiety,and depression of the patients before and after the intervention.The behavioral tasks of the detection task(DET),identification task(IDN),two back task(TWOB),the Groton maze learning task(GML),and international shopping list task(ISL)in the CogState software package were used to assess the cognitive processing speed,attention/vigilance,working memory,spatial problem-solving/error monitoring ability,and verbal learning and memory of the patients before and after the intervention.Results·Repeated measures ANOVA showed that the time effect[F=126.713,P＜0.001,partial η2(ηp2)=0.765]and interaction effect(F=7.080.P=0.011,ηp2=0.154)of the VAS scores in the two groups of patients were statistically significant.The time effect(F=9.114,P=0.004,ηp2=0.189),group effect(F=5.557,P=0.024,ηp2=0.125),and interaction effect(F=4.977,P=0.032,η2=0.113)of the TWOB score were all statistically significant.Only the time effects of BDI(F=45.273,P＜0.001,ηp2=0.578),BAI(F=31.432,P＜0.001,ηp2=0.473),GML(F=8.993,P=0.005,ηp2=0.209),and ISL(F=26.657,P＜0.001,ηp2=0.439)scores were statistically significant.There were no statistically significant effects of time,group,or interaction on the DET and IDN scores.Simple effect analysis showed that the VAS score of the real stimulation group was lower than that of the sham stimulation group after the intervention(F=8.805,P=0.005,ηp2=0.184),and the TWOB score of the real stimulation group was higher than that of the sham stimulation group(F=11.293,P=0.002,ηp2=0.225).Conclusion·Combining iTBS with routine inpatient treatment can enhance the efficacy of reducing alcohol craving in alcohol-dependent patients during the withdrawal period,and improve their working memory.

Objective:To summarize the best evidence for nutrition management of sarcopenia in patients undergoing maintenance hemodialysis (MHD), to guide the development of nutrition management programs.Methods:Using the 6S evidence model, literature on nutrition management of sarcopenia in MHD patients was electronically retrieved from databases and websites including UpToDate, Guidelines International Network, Joanna Briggs Institute Evidence-Based Health Care Center Database, European Society for Clinical Nutrition and Metabolism, UK Kidney Association, PubMed, Web of Science, China Biology Medicine disc, China National Knowledge Infrastructure, and Wanfang Data. The search period was from database establishment to July 30, 2024. After screening and quality assessment of the literature, evidence was extracted and summarized.Results:A total of 19 articles were included, comprising one clinical decision, six guidelines, five systematic reviews, five expert consensus, and two randomized controlled trials. Twenty-six pieces of evidence were summarized from six aspects of nutrition team establishment and counseling, nutritional screening and assessment, nutritional support, nutrient intake, nutritional monitoring, and health education.Conclusions:The evidence summary on nutrition management of sarcopenia in MHD patients provides a basis for implementing nutritional interventions. Evidence transformation and application should be conducted in accordance with patient preferences and the actual clinical context.

Objective To investigate the perioperative management of antithrombotic drugs in patients undergoing non-cardiac surgery.Methods Patients on long-term antithrombotic drugs who underwent non-cardiac surgery in our hospital were included.Through interviews with patients and physicians,perioperative antithrombotic medication regimens were reviewed and compared with the"Multidisciplinary Expert Consensus on Perioperative Management of Antithrombotic Therapy"to evaluate compliance with consensus and analyze influencing factors.Results A total of 372 patients were included in the analysis.Among them,355 patients were on long-term antiplatelet therapy alone,and 17 were on long-term oral anticoagulantion.364(97.8％)discontinued antithrombotic medications prior to surgery.109 patients(29.3％)received low molecular weight heparin(LMWH)bridging therapy.Among the 355 patients on antiplatelet therapy,108(30.4％)had discontinuation durations consistent with the consensus recommendations,while 186(52.4％)discontinued medications for longer periods.Postoperatively,the average hospital stay for antiplatelet therapy patients was 6.64 days,with only 37(10.4％)resuming therapy during hospitalization.The average hospital stay for patients on anticoagulants was 9.94 days,with only 2(11.8％)resuming therapy during hospitalization.Regarding perioperative risk assessment,only 40(10.8％)of patients underwent additional internal medical evaluation for thromboembolic risk after medication discontinuation,with the remainder assessed soly by surgeons.Coronary heart disease was an independent risk factor associated with internal medical evaluation(OR 2.851,95％CI 1.160-7.011,P=0.022).For bleeding risk assessment,surgeons evaluations aligned with the consensus in 68.0％of cases,but surgeons tended to underestimate risk compared to the consensus.Conclusions In this single-center study,perioperative antithrombotic management showed low compliance with expert consensus,characterized by prolonged preoperative medication discontinuation,high rates of LMWH bridging,and low postoperative in-hospital resumption of therapy.A robust multidisciplinary collaboration system should be established to enhance comprehensive patient assessment.