Objective: To investigate the damaging effects of red blood cell supernatant (RBC-S) stored for different durations (7 d, 14 d, and 28 d) on vascular endothelial cells, and to explore the underlying mechanisms using bioinformatics analysis, so as to provide references for optimizing red blood cell transfusion strategies. Methods: Human umbilical vein endothelial cells (HUVECs) were co-cultured with RBC-S stored for 7, 14 and 28 days, designated as the 7 d group, 14 d group and 28 d group respectively, which were collectively defined as the experimental groups. Cell damage was evaluated by cell proliferation assay (Cell Counting Kit8, CCK8), lactate dehydrogenase (LDH) release assay, 4′, 6diamidino2phenylindole (DAPI) staining, and flow cytometry for apoptosis and reactive oxygen species (ROS) levels. The damage degree of RBC-S on vascular endothelial cells was assessed by statistical analysis of damage data among different groups. Since the damage effect reached a plateau at all time points, the 28 d storage group was selected as the representative for further mechanistic studies. Transcriptomic analysis was performed to explore the role of frizzled class receptor 1 (FZD1) and Wnt signaling pathway in red blood cell storagerelated endothelial dysfunction. Results: Compared with the control group, the storage groups treated with 7 d, 14 d, and 28 d RBC-S showed significantly decreased cell proliferation rates [control group 100%, 7 d group (69.51±2.30)%, 14 d group (74.54±2.89)%, 28 d group (73.59±2.36)%, P<0.05], significantly reduced numbers of DAPI-stained cell nuclei [control group (213±12.5) per field, 7 d group (140.33±17.04) per field, 14 d group (152.00±23.72) per field, 28 d group (144.33±19.09) per field, P<0.05] and significantly increased LDH release [control group (1), 7 d group (8.33±1.41), 14 d group (9.23±0.83), 28 d group (9.16±0.60), P<0.05]. There was no significant difference in the degree of damage caused by RBC-S among different storage groups (P>0.05). With the prolongation of storage time, free hemoglobin (FHb) gradually increased [control group (not detected), 7 d (16.57±6.38) mg/L, 14 d (76.80±22.83) mg/L, 28 d (286.97±29.02) mg/L, P<0.05]. The apoptotic rate (20.53±2.94)% and ROS relative intensity (5.13±0.91) in the 28 d storage group were significantly higher than those in the control group (P<0.05). Transcriptomic analysis showed that FZD1 played a key role in vascular endothelial dysfunction induced by red blood cell storage and was closely related to the Wnt signaling regulatory network. Conclusion: RBC-S stored for 7 d, 14 d, or 28 d can all significantly damage vascular endothelial cells, and the damaging effect reaches a plateau at 7 d of storage. Mechanistic investigation of the 28 d group indicated that the downregulation of the FZD1/Wnt signaling pathway may play a critical role in vascular endothelial dysfunction induced by red blood cell storage, providing a theoretical basis for further optimizing red blood cell storage and transfusion strategies.

Immobilized enzyme-based enzyme electrode biosensors, characterized by high sensitivity and efficiency, strong specificity, and compact size, demonstrate broad application prospects in life science research, disease diagnosis and monitoring, etc. Immobilization of enzyme is a critical step in determining the performance (stability, sensitivity, and reproducibility) of the biosensors. Random immobilization (physical adsorption, covalent cross-linking, etc.) can easily bring about problems, such as decreased enzyme activity and relatively unstable immobilization. Whereas, directional immobilization utilizing amino acid residue mutation, affinity peptide fusion, or nucleotide-specific binding to restrict the orientation of the enzymes provides new possibilities to solve the problems caused by random immobilization. In this paper, the principles, advantages and disadvantages and the application progress of enzyme electrode biosensors of different directional immobilization strategies for enzyme molecular sensing elements by specific amino acids (lysine, histidine, cysteine, unnatural amino acid) with functional groups introduced based on site-specific mutation, affinity peptides (gold binding peptides, carbon binding peptides, carbohydrate binding domains) fused through genetic engineering, and specific binding between nucleotides and target enzymes (proteins) were reviewed, and the application fields, advantages and limitations of various immobilized enzyme interface characterization techniques were discussed, hoping to provide theoretical and technical guidance for the creation of high-performance enzyme sensing elements and the manufacture of enzyme electrode sensors.

The Chinese Pharmacopoeia is the legal technical standard which should be followed during the research, production, use, and administration of drugs. At present, the new edition of the Chinese Pharmacopoeia is planned to be promulgated and implemented. This article summarizes and analyzes the main characteristics and the content of updates and amendments of the Chinese Pharmacopoeia 2025 Edition(Volume Ⅰ), to provide a reference for the correct understanding and accurate implementation the new edition of the pharmacopoeia.

Objective To develop a deep learning model based on fundus retinal images to improve the detection rate of mild cognitive impairment(MCI)in patients with coronary heart disease,achieve early intervention and improve prognosis.Methods The study was a single-center cross-sectional study that retrospectively included patients diagnosed with coronary heart disease(CHD)by coronary angiography(≥50％ stenosis of at least one coronary vessel)from Beijing Anzhen Hospital between November 2021 and December 2022.The whole data set was randomly divided into the training set and the testing set according to the ratio of 8∶2 for model development.After that,the patient data of the same center from January 2023 to April 2023 were included in the time verification method to verify the model.The diagnostic criteria for MCI were MMSE＜27 or MoCA＜26.Four kinds of convolutional neural network(CNN)architectures were used to train fundus images,and a comprehensive vision model of MCI detection was established through model integration.The area under the curve(AUC),sensitivity and specificity of the receiver operating curve(ROC)were used to evaluate the performance of the AI model.Results We collected 5 880 eligible fundus images from 3 368 CHD patients.Based on the results of the MMSE scale,the algorithm was labeled,including 2 898 males and 527 MCI patients.The AUC of the deep learning model in the test group is 0.733(95％CI 0.688-0.778),and the sensitivity of the algorithm in the test group is 0.577(95％CI 0.528-0.625)by using the operating point with the maximum sum of sensitivity and specificity.With a specificity of 0.758(95％CI 0.714-0.802),corresponding to a validated AUC of 0.710(95％CI 0.601-0.818).Based on the results of the MoCA scale,the algorithm labels 2 437 males and 1 626 MCI patients.The AUC of the deep learning model in the test group was 0.702(95％CI 0.671-0.733).The operating point with the maximum sum of sensitivity and specificity was selected,and the sensitivity of the algorithm was 0.749(95％CI 0.719-0.778)and the specificity was 0.561(95％CI 0.527-0.595),corresponding to the AUC value of the verification group was 0.674(95％CI 0.622-0.726).Conclusions The deep learning algorithm model based on fundus images has good diagnostic performance,and may be used as a new non-invasive,convenient and rapid screening method for MCI in CHD population.

Aim Mouse model of myocardial hypertro-phy was established via intraperitoneal injection of iso-proterenol(ISO)in mice.This approach allows for an in-depth investigation into the pharmacological effects and mechanisms of action of emodin,offering novel in-sights and directions for the improvement of myocardial hypertrophy.Methods The mice were randomly di-vided into the following groups:control group(CON),emodin group(EMO),MAPK activator control group(EMO+Ani),model group(ISO),treatment group(ISO+EMO),and activator intervention group(ISO+EMO+Ani).After treatment with emodin and inter-vention with MAPK activator,the heart weight ratio and cardiac size of each group were observed.Hematoxy-lin-eosin(HE)staining was used to observe the patho-logical changes in cardiac tissue,and kits were utilized to measure the levels of GSH,LDH,and MDA in the serum.Western blot was employed to detect the protein expression levels of inflammatory and oxidative factors,as well as p-p38,p-ERK,p38,and ERK in cardiac tis-sue.Results Emodin can significantly inhibit the production of myocardial inflammatory and oxidative factors induced by ISO,thereby effectively alleviating the degree of myocardial hypertrophy and fibrosis.Af-ter the p38/ERK signaling pathway was specifically ac-tivated by farnesol,the improvement effect of emodin on myocardial hypertrophy was weakened.Further comparison revealed that,compared with the myocardi-al hypertrophy pathological model group,the pathologi-cal protein expression levels in the farnesol-treated group showed no significant difference,and were even higher in some indicators.Conclusion Emodin can effectively inhibit the release of inflammatory factors and improve the state of oxidative stress by modulating the p38/ERK signaling pathway,thereby exerting an ameliorative effect on myocardial hypertrophy.

Magnetism possesses both"force"and"field"properties,enabling non-contact interactions.Hepatopancreatobiliary(HPB)surgery is a field characterized by continuous innovation.In 2015,our team first proposed the concept of"magnetic surgery,"and subsequently advanced it from technical exploration to a disciplinary framework through organizing international conferences,founding a dedicated journal,and compiling textbooks.Many innovations originated in HPB practice:magnetic biliary and pancreatic anastomosis and magnetic recanalization for biliary strictures have significantly improved traditional approaches;magnetic vascular anastomosis has enabled rapid implantation of donor livers,supporting laparoscopic liver transplantation;and magnetic anchoring has provided multidimensional traction and exposure while reducing Trocar use and associated damage,thereby optimizing laparoscopic procedures.Looking ahead,multidisciplinary integration will drive the continued evolution of magnetic surgery,such as combining magnetic navigation with micro-nano surgical robots to achieve real-time tracking and guidance inside the body.These advances may overcome the limitations of conventional image guidance in applications like tumor ablation and catheter puncture.Ultimately,magnetic surgery is expected to evolve toward greater precision,minimal invasiveness,and intelligence,offering transformative solutions for HPB surgery and beyond.

Objective To evaluate the safety and efficacy of valve-in-valve transcatheter mitral valve replacement(ViV-TMVR)in patients with bioprosthetic valve degeneration who are at high surgical risk.Methods This study is a multi-center,retrospective cohort analysis of 20 consecutive patients who underwent transseptal ViV-TMVR using the Edwards SAPIEN 3 transcatheter heart valve(THV).The primary endpoints include technical success and procedural success,both defined according to the Mitral Valve Academic Research Consortium(MVARC)criteria,as well as mortality and functional change assessed based on New York Heart Association(NYHA)classification at 30-days and six months post-procedure.Clinical follow-up assessments are conducted at 30-days and six months.Results From February 2021 to October 2022,a total of 20 patients with symptoms of bioprosthetic valve degeneration were enrolled across nine sites in China.The patients had a mean age of(73.5±5.5)years,with 85.0％being females and 70.0％classified as NYHA class Ⅲ/Ⅳ.The study achieved a 100.0％technical success rate and a 90.0％procedural success rate finally.All patients remained alive during the 30-day follow-up period.However,six months post-intervention,two patients(10.0％)were re-hospitalized due to heart failure,and sadly,one of them(5.0％)died.None of the patients reported any adverse events related to ViV-TMVR during the follow-up period.Notably,there was a significant improvement in NYHA class compared to baseline(P=0.0004)at six-month follow-ups.Conclusions The transseptal ViV-TMVR technique proved to be highly successful and was associated with significant improvement in NYHA class function.These findings strongly suggest that it serves as a safe and efficient treatment alternative for high-risk patients suffering from bioprosthetic valve degeneration.

Magnetism possesses both"force"and"field"properties,enabling non-contact interactions.Hepatopancreatobiliary(HPB)surgery is a field characterized by continuous innovation.In 2015,our team first proposed the concept of"magnetic surgery,"and subsequently advanced it from technical exploration to a disciplinary framework through organizing international conferences,founding a dedicated journal,and compiling textbooks.Many innovations originated in HPB practice:magnetic biliary and pancreatic anastomosis and magnetic recanalization for biliary strictures have significantly improved traditional approaches;magnetic vascular anastomosis has enabled rapid implantation of donor livers,supporting laparoscopic liver transplantation;and magnetic anchoring has provided multidimensional traction and exposure while reducing Trocar use and associated damage,thereby optimizing laparoscopic procedures.Looking ahead,multidisciplinary integration will drive the continued evolution of magnetic surgery,such as combining magnetic navigation with micro-nano surgical robots to achieve real-time tracking and guidance inside the body.These advances may overcome the limitations of conventional image guidance in applications like tumor ablation and catheter puncture.Ultimately,magnetic surgery is expected to evolve toward greater precision,minimal invasiveness,and intelligence,offering transformative solutions for HPB surgery and beyond.

Aim To investigate the effect of circRNF13 on oxaliplatin resistance in colorectal cancer cells and its related mechanism.Methods Cell transfection was used to overexpress circRNF13 in oxaliplatin-sensi-tive colorectal cancer cells SW620 or to inhibit cir-cRNF13 expression in oxaliplatin-resistant colorectal cancer cells SW620/OXA.Real-time quantitative fluo-rescent PCR(qRT-PCR)was used to detect the ex-pression levels of circRNF13,miR-16-5p,and mir-324-5p.Cell Count Kit 8(CCK-8)was used to meas-ure cell viability.Clonal formation experiments were used to measure clonal formation number.Western blot was used to detect CyclinD1,PCNA,Bax,Bcl-2 and cleaved caspase-3 protein expression level.Flow cy-tometry and in situ end labeling(TUNEL)were used to detect apoptosis.Dual luciferase assay was used to verify the targeting relationship between circRNF13 and miR-16-5p and mir-324-5p.Results The expression level of circRNF13 in SW620/OXA cells was signifi-cantly higher than that in SW620 cells,while the ex-pression levels of miR-16-5p and miR-324-5p were sig-nificantly lower than those in SW620 cells(P＜0.05).Overexpression of circRNF13 significantly in-creased the IC50 of oxaliplatin against SW620 cells,and inhibition of circRNF13 expression significantly decreased the IC50 of oxaliplatin against SW620/OXA cells.Overexpression of circRNF13 significantly in-creased cell viability,clonal formation number,Cy-clinD1,PCNA and Bcl-2 levels of oxaliplatin treated SW620 cells,while significantly decreased apoptosis rate,apoptosis index,Bax and cleaved caspase-3 lev-els(P＜0.05).Inhibition of circRNF13 expression significantly decreased the cell viability,clonal forma-tion number,CyclinD1,PCNA and Bcl-2 levels of ox-aliplatin treated SW620/OXA cells,while significantly increased the apoptosis rate,apoptosis index,Bax and cleaved caspase-3 levels(P＜0.05).circRNF13 tar-geted inhibition of the expression of miR-16-5p and mir-324-5p.Conclusions circRNF13 can increase oxaliplatin resistance in colorectal cancer cells,and the mechanism may be related to the targeted inhibition of circRNF13 on the expression of miR-16-5p,mir-324-5p and other miRNAs.

Aim To investigate the effect of circRNF13 on oxaliplatin resistance in colorectal cancer cells and its related mechanism.Methods Cell transfection was used to overexpress circRNF13 in oxaliplatin-sensi-tive colorectal cancer cells SW620 or to inhibit cir-cRNF13 expression in oxaliplatin-resistant colorectal cancer cells SW620/OXA.Real-time quantitative fluo-rescent PCR(qRT-PCR)was used to detect the ex-pression levels of circRNF13,miR-16-5p,and mir-324-5p.Cell Count Kit 8(CCK-8)was used to meas-ure cell viability.Clonal formation experiments were used to measure clonal formation number.Western blot was used to detect CyclinD1,PCNA,Bax,Bcl-2 and cleaved caspase-3 protein expression level.Flow cy-tometry and in situ end labeling(TUNEL)were used to detect apoptosis.Dual luciferase assay was used to verify the targeting relationship between circRNF13 and miR-16-5p and mir-324-5p.Results The expression level of circRNF13 in SW620/OXA cells was signifi-cantly higher than that in SW620 cells,while the ex-pression levels of miR-16-5p and miR-324-5p were sig-nificantly lower than those in SW620 cells(P＜0.05).Overexpression of circRNF13 significantly in-creased the IC50 of oxaliplatin against SW620 cells,and inhibition of circRNF13 expression significantly decreased the IC50 of oxaliplatin against SW620/OXA cells.Overexpression of circRNF13 significantly in-creased cell viability,clonal formation number,Cy-clinD1,PCNA and Bcl-2 levels of oxaliplatin treated SW620 cells,while significantly decreased apoptosis rate,apoptosis index,Bax and cleaved caspase-3 lev-els(P＜0.05).Inhibition of circRNF13 expression significantly decreased the cell viability,clonal forma-tion number,CyclinD1,PCNA and Bcl-2 levels of ox-aliplatin treated SW620/OXA cells,while significantly increased the apoptosis rate,apoptosis index,Bax and cleaved caspase-3 levels(P＜0.05).circRNF13 tar-geted inhibition of the expression of miR-16-5p and mir-324-5p.Conclusions circRNF13 can increase oxaliplatin resistance in colorectal cancer cells,and the mechanism may be related to the targeted inhibition of circRNF13 on the expression of miR-16-5p,mir-324-5p and other miRNAs.