Objective To construct a targeted drug delivery system, Ang-BEVs@Dox, based on Angiopep-2 peptide-modified bacterial extracellular vesicles （BEVs） loaded with doxorubicin （Dox）, overcome the challenges of blood-brain barrier （BBB） penetration and systemic toxicity in chemotherapy for glioblastoma （GBM）, enhance drug targeting to brain tumors and reduce its toxic side effects. Methods BEVs derived from Escherichia coli were isolated using ultracentrifugation. The targeting ligand Angiopep-2, specific for the LRP-1 receptor, was conjugated onto the surface of BEVs to construct the targeted carrier （Ang-BEVs）. Dox was loaded into Ang-BEVs using low-frequency sonication to form Ang-BEVs@Dox. The physicochemical properties （morphology and size） of the carriers were characterized by transmission electron microscopy （TEM） and dynamic light scattering （DLS）. The BBB-penetrating capability, in vitro/in vivo anti-tumor efficacy, and biosafety of the system were evaluated using cellular uptake assays, 3D tumor spheroid models, and orthotopic tumor-bearing mouse models. Results ① Carrier characterization and in vitro efficacy: Ang-BEVs@Dox exhibited a particle size of approximately 100 nm and maintained structural stability after Dox loading. It significantly enhanced cellular uptake efficiency in U87MG cells and achieved deep penetration within 3D tumor spheroids. Cytotoxicity assays demonstrated synergistic anti-tumor effects between the BEVs and Dox in the Ang-BEVs@Dox system. ② In vivo targeting and anti-tumor efficacy: In orthotopic tumor-bearing mouse models, Ang-BEVs@Dox effectively penetrated the BBB and significantly inhibited tumor growth, extending the median survival time of tumor-bearing mice to 33.5 days （compared to 23.5 days in the blank control group, P<0.001）. Immunohistochemical analysis revealed significant suppression of the tumor cell proliferation marker Ki-67 and enhancement of the apoptosis marker TUNEL staining signals. ③ Biosafety: Major organs from mice in the Ang-BEVs@Dox treatment group showed no observable pathological damage, indicating good biosafety. Conclusion This study successfully constructed an Angiopep-2 peptide-modified engineered BEVs delivery system （Ang-BEVs@Dox）. Through Angiopep-2-mediated BBB penetration and tumor targeting, it significantly enhanced the accumulation and therapeutic efficacy of BEVs at the GBM site. This method combined efficient delivery, low systemic toxicity, and clinical translation potential, which provided an innovative solution to overcome the therapeutic bottleneck in GBM treatment.

Objective To establish and verify a mature and stable glioblastoma(GBM)organoid model,so as to provide an accurate and personalized preclinical model for the research and treatment of GBM.Methods Fresh GBM tissues obtained through surgical procedures were initially processed,and then GBM stem cells(GSCs)were isolated using stem cell culture medium and were identified.Subsequently,GSCs were cultured in organoid culture medium for 3D cultivation,and GBM organoids were successfully obtained.The histological morphology of GBM organoids was observed by hematoxylin-eosin(H-E)staining;the stemness and similarity to the parental tumor were identified by immunofluorescence staining;and the in vivo tumorigenic ability of GBM organoids was identified by orthotopic tumorigenesis experiments in nude mice.Results A total of 7 GBM organoids were constructed from 9 human GBM samples,with a morphology resembling"neurosphere",and the average duration for organoid formation was 1 week.H-E staining results showed that the histological morphology of GBM organoids under high-power microscope was very similar to that of GBM tumor tissues;immunofluorescence staining results indicated that the GBM organoids possessed stemness characteristics and histological cellular similarity;and GBM organoids had a stronger tumorigenic ability compared to ordinary GBM cells in nude mice.Conclusion This study presents a stable and reliable method for constructing GBM organoids retaining the histological characteristics of the original GBM tissue,which providing new insights for future GBM research and clinical practice.

WD repeat domain 1(WDR1)is a highly conserved cytoskeleton-associated protein that plays a crucial role in physiological processes such as actin cytoskeleton remodeling,dynamic regulation of intercellular junctions,cell division,and migration.WDR1 exhibits abnormal high ex-pression in various malignant tumors,including breast cancer,ovarian cancer,and thyroid cancer,and has been demonstrated to significantly promote the invasive and migratory capabilities of tumor cells,suggesting its important role in the malignant progression of tumors.Moreover,the expression level of WDR1 is closely related to the clinical prognosis of patients with multiple malignant tumors.Especially in patients with esophageal cancer and osteosarcoma,its high expression often indicates a poor overall survival rate.WDR1 can promote tumor initiation and progression by regulating the Wnt/β-Catenin signaling pathway and the Hippo-YAP signaling pathway.Meanwhile,its expression is also subject to multi-level regulation by transcription activation factors and long non-coding RNAs(lncR-NAs),thereby influencing the proliferation,migration,and other biological behaviors of tumor cells.Additionally,WDR1 can further drive the invasive growth and metastatic potential of tumors by regu-lating the epithelial-mesenchymal transition(EMT)process.This article aimed to systematically re-view the research progress in recent years regarding the biological functions and molecular mechanisms of WDR1 in tumor initiation and development,with a view to providing new theoretical foundations and research directions for the early diagnosis,prognosis assessment,and individualized treatment of clinical tumors.

Objective:To investigate the value of histological evaluation in predicting endoscopic relapse among patients with ulcerative colitis (UC) who were in endoscopic remission, and to compare the usefulness of various histological scoring systems.Methods:Histological sections from 61 patients with UC who were in endoscopic remission were retrospectively analyzed, at Peking University Third Hospital, Beijing, China from January 2015 to June 2021. They were subdivided into endoscopic persistent remission group (remission group, n=31, Mayo endoscopic score 0) and endoscopic relapse group (relapse group, n=30, Mayo endoscopic score≥1) according to the results of the first endoscopic reexamination after the biopsy. Histological evaluation was performed using the Geboes score (GS) and its simplified version (SGS), the Nancy index (NI) and the Robarts histopathological index (RHI). The median and maximum histological scores for each case in all biopsies were recorded. Univariate comparisons were performed using chi-squares and multivariate analysis using binary logistic regression. The values of four histological evaluation systems for predicting endoscopic relapse among UC patients in endoscopic remission were analyzed using receiver operating characteristic (ROC) curves. Results:Significant differences were observed between the remission and relapse groups. The differences were more pronounced in the maximum histological scores; the mean and highest results of area under the ROC curve scores (AUC) for GS, SGS, NI, and RHI were 0.657, 0.668, 0.682, 0.691, and 0.866, 0.863, 0.864, 0.869, respectively. The differences were statistically significant ( P<0.05). The corresponding best cut-offs were GS≥2B.1, SGS≥2B.1, NI≥2, and RHI≥2.5, respectively, which meant mild active inflammation histologically, while there was no statistical difference of AUC among the four histological scoring indices ( P>0.05). Univariate and multivariate analyses revealed statistically significant differences in the number of neutrophils in the epithelium and lamina propria ( P<0.05). Conclusions:Biopsies from UC patients in endoscopic remission may still have histological active inflammation which appears to correlate with endoscopic relapse. Four commonly used histological scoring systems can be used to assess the risk of endoscopic relapse among UC patients in endoscopic remission. The patients who more likely have endoscopic relapse seem to have a histological score greater than the cut-off value (i.e., mild histological activity). The maximum histological scores can accurately predict the risk of endoscopic relapse, while the presence of epithelial and laminar propria neutrophil infiltrates can independently predict the endoscopic relapse in these patients. Considering the utility and convenience in routine practice, NI is recommended for evaluating histological inflammatory activity.

In order to study the effect of antimicrobial drug reduction on the resistance of E.coli and diversity of antibiotic resistance genes(ARGs)and mobile genetic elements(MGEs)in the intes-tinal microbiota of dairy cows in dairy farms in Hebei Province,and to evaluate the implementation effect of antimicrobial drug reduction actions,anal swab samples and fecal samples were collected from three qualified dairy farms and two non-standard dairy farms in different regions of Hebei Province,and E.coli was isolated from the anal swab samples and tested for their resistance to 16 antimicrobial drugs,and the differences in the drug resistance rate of E.coli in the two types of dairy farms were analyzed.SYBR fluorescent dye method was used to detect ARGs and MGEs in dairy feces,their relative abundance was calculated,the differences and correlation between the two types of dairy farms were analyzed.The results showed that a total of 192 ARGs and 7 MGEs were detected in 15 samples,and the relative abundance of ARGs was the highest in β-lactam resistance genes,followed by tetracycline resistance genes.The detection of ARGs and MGEs in some stand-ard dairy farms was lower than that in non-standard farms,suggesting that the large use of antimi-crobial drugs may be related to the production and transmission of ARGs,and the number of MG-Es detected was correlated with the number of ARGs detected,and ARGs may be transmitted through horizontal gene transfer.The results showed that there was little difference in the drug re-sistance rate of E.coli from the two types of dairy farms,and multi-drug resistant strains were more likely to occur in non-standard farms.The implementation of antimicrobial reduction had a certain effect on the diversity of antibiotic resistance genes,but had no significant effect on the prevalence of E.coli drug resistance and the diversity of ARGs in dairy farms in the short term,and there was little difference between different regions,which also revealed the prevalence of anti-biotic resistance in dairy farms.

Bromodomain-containing protein 4(BRD4),a pivotal member of the bromodomain and extra-ter-minal domain(BET)family,is integral to the regulation of vital biological processes,including the cell cycle and gene ex-pression.Studies have identified extensive expression of BRD4 in cardiomyocytes and its influence on various signaling pathways.The dysregulation of BRD4 leads to significant alterations in these pathways,culminating in pathological states such as myocardial inflammation,fibrosis,oxidative stress,and hypertrophy.These changes are instrumental in the onset and progression of cardiovascular diseases.This review summarizes the advances in research on BRD4 and its inhibitors in the context of cardiovascular diseases,with a focus on providing insights for precise therapeutic interventions.

Atrial fibrillation(AF),a prevalent clinical tachyarrhythmia,has seen an increase in incidence with advancing age over recent decades.AF leads to severe complications,including stroke and heart failure,and is a sig-nificant cause of mortality.The pathogenesis of AF involves critical factors such as atrial electrical and structural remodeling,oxidative stress,and inflammatory responses.Recent studies highlight the pivotal role of macrophages in the initiation and progression of AF.This study focuses on the contributions of macrophages to the mechanisms of AF and summarizes cur-rent research findings.

AIM:To develop and validate a predic-tive model for the risk of high plasma concentra-tion of voriconazole,and to guide clinical individual-ized medication of voriconazole.METHODS:Based on the real-world data from the hospital Informa-tion system(HIS),the clinical data of hospitalized patients who received voriconazole treatment and underwent voriconazole plasma concentration monitoring in our hospital from August 2017 to Au-gust 2021 were collected.Univariate and multivari-ate logistic regression analysis were performed on the included influencing factors.At the same time,in order to minimize the potential collinearity and overfitting between variables,the least absolute shrinkage and selection operator regression were used to screen the potential predictors.Logistic re-gression analysis was used to construct a predic-tion model for the risk of high plasma concentra-tion of voriconazole.C-index,calibration chart and clinical decision curve analysis were used to evalu-ate the discrimination,consistency and clinical ap-plicability of the model,and a nomogram was drawn.RESULTS:A total of 147 patients were en-rolled in this study.Plasma albumin and procalcito-nin were selected as predictive variables for Logis-tic regression analysis,and the prediction model was established.Draw predict voriconazole nomo-gram risk blood drug concentration on the high side.The receiver operating characteristic curve showed that the AUC of the prediction model for predicting the risk of high plasma concentration of voriconazole was 0.787(95%CI 0.663-0.911).Vori-conazole blood drug concentration was high inci-dence of cut-off value was 33.06%,sensitivity was 63.64%,87.65%and 58.33%positive predictive val-ue,negative predictive value of 89.87%.The cali-bration curve showed good consistency,and the clinical decision curve showed that the model had a positive net benefit when the threshold probabili-ty was between 6.67%and 99.99%.CONCLUSION:The predictive model for the risk of high plasma concentration of voriconazole has good predictive efficacy,which can provide guidance for clinical in-dividualized medication of voriconazole.

Objective:To observe the effect of high glucose downregulated microRNA(miR)-99a on hepatic sinus dysfunction and metformin intervention, and to explore the pathogenesis of diabetes-induced fatty liver and possible mechanism of metformin.Methods:The cultured human liver sinusoidal endothelial cells were randomly divided into normal control group, high glucose model group, miR-99a overexpression group, miR-99a overexpression negative control group, insulin-like growth factor 1 receptor(IGF-1R) inhibitor group, mammalian target of rapamycin(mTOR) inhibitor group, and metformin treatment group. The mRNA expressions of miR-99a were detected with realtime quantitative PCR(RT-qPCR), and the expression levels and distribution of IGF-1R, phosphorylated(p-)mTOR and vitronectin(VN) were detected by Western blotting and immunofluorescence. The ultrastructure of human liver sinusoidal endothelial cells was observed using scanning electron microscope.Results:Compared with normal control group, the mRNA expression of miR-99a was downregulated( P=0.008), while the protein expressions of IGF-1R, mTOR, and VN were significantly increased, and the diameter and number of fenestrae decreased significantly in high glucose model group. Compared with high glucose model group, after the treatment with metformin, the mRNA expression of miR-99a was upregulated, while the protein expressions of IGF-1R, mTOR, and VN were significantly decreased( P=0.001, P=0.016, P=0.005, respectively), the number of fenestras increased and the diameter became larger in miR-99a overexpression group, IGF-1R inhibitor group, mTOR inhibitor group, and metformin treatment group. After overexpression of miR-99a, the protein expressions of IGF-1R, p-mTOR, and VN were significantly reduced( P=0.007, P=0.013, P=0.003, respectively); After administration of IGF-1R inhibitors, the expressions of p-mTOR and VN significantly decreased( P=0.006, P=0.009, respectively), following treatment with the mTOR inhibitor, the expression of VN was significantly reduced( P=0.008), while the expression of IGF-1R remained unchanged( P=0.553). Conclusions:Downregulating of miR-99a with high glucose induced hepatic sinus dysfunction, which may be related to the regulation of IGF-1R/mTOR pathway. Metformin increased the expression of miR-99a, thereby inhibiting high glucose-induced hepatic sinusoidal dysfunction.

The condition of patients with severe traumatic brain injury (sTBI) complicated by corona virus 2019 disease (COVID-19) is complex. sTBI can significantly increase the probability of COVID-19 developing into severe or critical stage, while COVID-19 can also increase the surgical risk of sTBI and the severity of postoperative lung lesions. There are many contradictions in the treatment process, which brings difficulties to the clinical treatment of such patients. Up to now, there are few clinical studies and therapeutic norms relevant to sTBI complicated by COVID-19. In order to standardize the clinical treatment of such patients, Critical Care Medicine Branch of China International Exchange and Promotive Association for Medical and Healthcare and Editorial Board of Chinese Journal of Trauma organized relevant experts to formulate the Chinese expert consensus on clinical treatment of adult patients with severe traumatic brain injury complicated by corona virus infection 2019 ( version 2023) based on the joint prevention and control mechanism scheme of the State Council and domestic and foreign literatures on sTBI and COVID-19 in the past 3 years of the international epidemic. Fifteen recommendations focused on emergency treatment, emergency surgery and comprehensive management were put forward to provide a guidance for the diagnosis and treatment of sTBI complicated by COVID-19.