BackgroundMonitoring the blood concentrations of antipsychotic drugs and their metabolites can guide the adjustment of clinical treatment plans， improving therapeutic efficacy while reducing adverse effects. However， there is currently a lack of a method that can accurately and efficiently quantitatively detect multiple antipsychotic drugs and their metabolites. ObjectiveTo establish a ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method for the simultaneous identification and quantitation of 12 antipsychotic drugs and their main metabolites in human serum. MethodsUsing UPLC-MS/MS technology， protein precipitation method was employed for sample pretreatment. An Agela Technologies Durashell C₈ chromatographic column （50 mm×3.0 mm， 5 μm） was selected for chromatographic separation with gradient elution. The flow rate was 0.4 mL/min， and the total analysis time was 5 minutes. The column temperature was 40℃. The mass spectrometry detection was carried out in the multiple reaction monitoring （MRM） mode， and the isotope internal standard method was used for quantification. ResultsThe relative standard deviation （RSD） of the internal standard normalization matrix effect factor for 12 antipsychotic drugs and their main metabolites at low and high quality concentrations was all less than 15%. The extraction recovery rate was 85% to 115%. They showed good linear relationships within their respective standard curve ranges （r>0.995）. At low， medium， and high quality concentrations， the accuracy was 85.24% to 114.71%， and the RSD of intra-batch and inter-batch precision was all ≤14.15%， with good stability. ConclusionAll the analytical performance indicators of this method meet the verification requirements， providing an analytical means for the quantitative detection of antipsychotic drugs and their main metabolites in human serum. ［Funded by The Third Batch of Science and Technology Projects in Chaozhou City in 2023 （number， 202303GY02）］

Objective:To evaluate the efficacy of digitally assisted design of surgical template for fibular composite tissue flap in reconstruction of segmental mandibular defect with soft tissue defect.Methods:A retrospective analysis was conducted on 30 patients who were treated at the Department of Otolaryngology Head and Neck Surgery, the 920th Hospital of the Joint Logistics Support Force of Chinese PLA from July 2020 to May 2024 for segmental mandibular defects combined with soft tissue defect. The cohort comprised 19 males and 11 females and aged 11-71 years (44.33 years±15.31 years). Pathological diagnoses of the patients were ameloblastoma (14 cases), squamous cell carcinoma (9 cases), osteomyelitis (4 cases), and odontogenic keratocyst (3 cases). Primary reconstructive surgery were performed on 21 patients after surgical resection of lesions, and 9 patients received secondary reconstructive surgery. The length of mandibular defect ranged from 75.83 mm to 111.45 mm (87.31 mm±12.00 mm), and soft tissue defects were measured at 5.0 cm×1.8 cm to 8.6 cm×2.1 cm (mean area 13.63 cm 2±2.42 cm 2). Preoperative CTA was performed to locate the perforator of peroneal artery and for design of digital surgical template. Intraoperatively, a modified fibula composite tissue flap harvesting technique was employed and that involved in: CTA-guided perforator planning, fibula osteotomy, anterior intermuscular septum exposure for identifying the peroneal artery, and a digital template-assisted fibula crafting for reconstruction of mandibule and soft tissue defect. Postoperative follow-ups were conducted at 1, 3, and 6 months, followed by quarterly reviews at outpatient clinic or via telephone interviews. Statistical analysis was performed using SPSS 27.0 software with descriptive statistical methods. Results:After surgery, all the fibula composite tissue flaps were viable and the incision wound healed well. Two patients had partial necroses at distal edge of the flaps, and they were healed after treatment. One patient had donor site infection and healed after anti-infective treatment. One month after the surgery, patients were assessed according to the recovery of face, mouth opening and occlusion, of which 25 patients (83.3%) were rated of Grade I, 4 (13.3%) of Grade Ⅱ, and 1 (3.4%) of Grade Ⅳ, with an excellent and good rates of 96.6%. The average distance of condylar movement on the affected side was 1.28 mm±0.35 mm. Postoperative follow-up lasted for 10 to 22 months, with 19.17 months±2.14 months in average. Assessment at the final follow-up was found that a total of 26 patients (86.8%) were of Grade I, 3 (10.0%) of Grade Ⅱ, and 1 (3.3%) of Grade Ⅳ and all the transferred fibula showed good alignment with the mandible over the postoperative follow-up period.Conclusion:Digital template-assisted design of fibular composite tissue flap enables a precise vascular protection, individualised osteotomy and functional restoration in reconstruction of segmental mandibular defect with soft tissue defect. It demonstrates a high clinical feasibility.

With the rapid development of generative artificial intelligence(GAI)technologies,their widespread application in academic research and writing is continuously expanding the boundaries of sci-entific inquiry.However,this trend has also raised a series of ethical and regulatory challenges,inclu-ding issues related to authorship,content authenticity,citation accuracy,and accountability.In light of the growing involvement of AI in generating academic content,establishing an open,controllable,and trustworthy ethical governance framework has become a key task for safeguarding research integrity and maintaining trust within the academic community.This expert consensus outlines ethical requirements across key stages of AI-assisted academic writing-including topic selection,data management,citation practices,and authorship attribution.It aims to clarify the boundaries and ethical obligations surrounding AI use in academic writing,ensuring that technological tools enhance efficiency without compromising in-tegrity.The goal is to provide guidance and institutional support for building a responsible and sustainable research ecosystem.

OBJECTIVE To explore the effectiveness and possible mechanism of Shangke Lengtong Patch in treating knee osteo-arthritis with cold-dampness arthralgia obstruction.METHODS A total of 70 patients who met the inclusion criteria of knee osteoar-thritis with cold-dampness arthralgia obstruction in the Orthopedics Department of Affiliated Hospital of Nanjing University of Chinese Medicine from November to December 2024 were randomly divided into an experimental group and a control group,with 35 cases in each group.During the treatment,1 case dropped out of the experimental group,3 cases dropped out of the control group,and 1 case was discontinued.The experimental group was treated with Shangke Lengtong Patch,and the control group was treated with Compound Nanxing Zhitong Ointment.The WOMAC scores and TCM syndrome scores of the two groups before and after treatment were compared to comprehensively evaluate the clinical efficacy.The changes in the expression levels of CGRP,substance P,HMGB1,IL-1β,CX-CL12,and CXCR4 in the serum of the two groups were detected by ELISA.RESULTS After 3,7,and 14 d of treatment,the WOMAC scores and TCM syndrome scores of the two groups were significantly reduced(P<0.05,P<0.01),and the score of aggrava-ted cold in the experimental group was better than that in the control group at 7 d of treatment(P<0.05);after 14 d of treatment,the expression levels of CGRP,substance P,HMGB1,IL-1β,CXCL12,and CXCR4 in the serum of the two groups were significantly re-duced(P<0.05),and there was no statistical difference between the two groups.CONCLUSION Shangke Lengtong Patch can sig-nificantly relieve the pain symptoms of knee osteoarthritis patients with cold-dampness arthralgia obstruction and improve the joint func-tion of patients.It may improve synovial inflammation by inhibiting the HMGB1/CXCL12/CXCR4 pathway,thereby exerting a thera-peutic effect.

Objective To evaluate the diagnostic performance of the minimum-cost-path-based CT angiography-derived fractional flow reserve(MCP-FFR)and the deep learning-driven CT angiography-derived fractional flow reserve(DeepCL-FFR),and to particularly explore the potential value of the DeepCL algorithm in improving diagnostic accuracy within the"gray zone."Methods A retrospective analysis was conducted on 151 coronary vessels from 109 patients with coronary artery disease,who were hospitalized at the General Hospital of the People's Liberation Army between January 2020 and June 2021.Pearson correlation and Bland-Altman plots were employed to assess the correlation and agreement of the two CT-FFR methods with invasive FFR.A CT-FFR range of 0.70-0.80 was defined as the diagnostic"gray zone."The accuracy,sensitivity,specificity,positive predictive value,and negative predictive value for detecting hemodynamic abnormalities were calculated and analyzed.The DeLong test was used to compare the areas under the receiver operating characteristic curves(AUC)between the two CT-FFR calculation methods.Results Both CT-FFR methods exhibited a positive correlation with invasive FFR(MCP-FFR:r=0.75,P＜0.001;DeepCL-FFR:r=0.86,P＜0.001)and showed good agreement(MCP-FFR:mean difference=0.010,P=0.351;DeepCL-FFR:mean difference=-0.003,P=0.772).Both DeepCL-FFR(AUC 0.97,95％CI 0.94-0.99)and MCP-FFR(AUC 0.92,95％CI 0.88-0.97)demonstrated favorable diagnostic performance for detecting hemodynamic abnormalities(P=0.122).In the"gray zone"for hemodynamic abnormality,the diagnostic accuracy of MCP-FFR was 68.8％,whereas DeepCL-FFR increased it to 89.7％.DeepCL-FFR also exhibited superior diagnostic performance(AUC 0.89,95％CI 0.73-0.99)within the"gray zone,"which was significantly higher than that of MCP-FFR(AUC 0.71,95％CI 0.54-0.87)(P＜0.001).Conclusions The deep learning-driven coronary centerline extraction algorithm,DeepCL,demonstrates superior diagnostic performance in CT-FFR for detecting hemodynamic abnormalities,particularly by significantly improving diagnostic accuracy in the"gray zone."

Objective To investigate the regulatory effects and molecular mechanisms of extracellular vesicles(EVs)secreted by adri-amycin(ADR)-resistant breast cancer cells on vascular endothelial cells related to tumor microenvironment.Methods The ADR-sen-sitive breast cancer cell line(MCF-7)and its ADR-resistant counterpart(MCF-7-ADR)were established in vitro.Then,the EVs were isolated from the supernatant of cell culture using a commercial kit.The fluorescence labeling and tracking technology using PKH67 was used to validate the internalization of EVs by human umbilical vein endothelial cell(HUVEC).The effects of EVs on the proliferation,migration,and angiogenesis of HUVEC were assessed by the colony formation,scratch wound healing,and tube formation assays,re-spectively.The expression level of CD31 protein was detected by Western blot,and the levels of vascular endothelial growth factor(VEGF)in cell supernatant were determined by ELISA.The microRNA(miRNA)sequencing technology was utilized to identify the differentially expressed miRNAs in EVs secreted by ADR-resistant cells.Results Compared with the ADR-sensitive strain,the EVs secreted by the ADR-resistant strain significantly enhanced the proliferation(colony number increased by 38.7%,P=0.011),migra-tion(scratch healing rate improved by 42.3%,P=0.004),and tube formation ability(tube formation rates increased by 1.8 times and 2.3 times at 4 h and 16 h,respectively,P=0.044 and 0.010)of HUVEC.Western blot analysis showed that the expression level of CD31 in the EVs secreted by the ADR-resistant strain increased by 1.85 times compared with the ADR-sensitive strain(P=0.039).The results of ELISA showed that the expression level of VEGF increased by 2.12 times(P=0.012).The miRNA sequencing identified a differentially expressed miRNA profile.Conclusion The ADR-resistant breast cancer cells significantly enhance the proliferation,migration,and angiogenesis of vascular endothelial cells by secreting EVs rich in specific miRNA,and its mechanism may be related to the up-regulation of CD31/VEGF protein levels mediated by EVs,suggesting that the drug-resistant cells reshaping the tumor vascular microenvironment through EVs may become a new therapeutic target of chemotherapy resistance in breast cancer.

Objective To investigate the expression and clinical significance of S100 cabinding protein A7(S100A7),insulin-like growth factor-1(IGF-1),and receptor for advanced glycation end products(RAGE)in type Ⅰ endometrial carcinoma(EC).Methods 60 patients with type 1 EC and 30 normal endometrial tissues were selected and analyzed by real-time fluorescent quantita-tive polymerase chain reaction(RT-qPCR).RT-qPCR was used to detect the expression of S100A7,IGF-1 and RAGE mRNA in dif-ferent endometrial tissues.The protein expression of S100A7,IGF-1 and RAGE in 107 type Ⅰ EC tissues,24 cases endometrial tissues with atypical hyperplasia and 30 normal endometrial tissues were detected by immunohistochemical method.The correlation between the three and the clinicopathological characteristics of the patients was analyzed.Kaplan-Meier method was used to draw the survival curve,and COX regression analysis was performed to analyze the factors affecting the survival and prognosis of patients.Results The expres-sions of S100A7,IGF-1 and RAGE mRNA in EC tissues were increased compared with normal endometrial tissues(P＜0.05).The positive rates of S100A7,IGF-1 and RAGE proteins increased with the progression of endometrial lesions,and there was a significant positive correlation among them(P＜0.05).For different International Federation of Gynecology and Obstetrics(FIGO)staging,pres-ence or absence of lymph node metastasis,and degree of tissue differentiation in type Ⅰ EC,The positive rates of S100A7,IGF-1 and RAGE were different(P＜0.05).Kaplan-Meier survival analysis showed that the average survival time of patients with positive expres-sion of S100A7,IGF-1 and RAGE was lower.COX regression analysis showed that lymph node metastasis,high expression of S100A7,IGF-1 and RAGE were independent risk factors for prognosis of patients with type Ⅰ EC.Conclusion The increased expression levels of S100A7,IGF-1 and RAGE in type Ⅰ EC are closely associated with the emergence,progress and bleak prognosis of EC.

BACKGROUND:Synovitis is involved in all stages of osteoarthritis and is a key factor contributing to the development of osteoarthritis.Studies have shown that circular RNA(circRNA)plays an important role in the proliferation,apoptosis and extracellular matrix degradation of synovial cells and chondrocytes.OBJECTIVE:To observe the effects of circRNA SEC24A on the interleukin-1β-induced proliferation,apoptosis,and expression of inflammatory factors in human synovial fibroblasts.METHODS:Human synovial fibroblasts were divided into four groups,including control group,interleukin-1β group,empty vector group,and sh-circSEC24A group.Except for the control group,the other three groups were induced with 10 ng/mL interleukin-1β for 24 hours to establish inflammatory cell models;the empty vector group and sh-circSEC24A group were infected with empty vector virus and lentiviral vector knocking down circSEC24A.CCK-8 assay was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.ELISA was used to detect the levels of matrix metalloproteinase-9,matrix metalloproteinase-13,interleukin-6,and tumor necrosis factor-α in cell supernatant.Western blot assay was used to detect the relative expression levels of Bax,Bcl-2,matrix metalloproteinase-9,matrix metalloproteinase-13,casepase3,cleaved-casepase3,casepase8,and cleaved-casepase8 proteins in cells.RESULTS AND CONCLUSION:(1)qRT-PCR results showed that compared with the normal group,the expression of circSEC24A in human synovial fibroblasts induced by interleukin 1β was significantly up-regulated.(2)The absorbance value of cells in the sh-circSEC24A group detected by CCK-8 assay was significantly higher than that of interleukin 1β group and empty vector group(P＜0.05).The apoptosis rate of sh-circSEC24A group detected by flow cytometry was significantly lower than that of interleukin 1β group and empty vector group(P＜0.05).(3)The levels of tumor necrosis factor α and interleukin 6 in the supernatant of human synovial fibroblasts in the sh-circSEC24A group detected by ELISA were significantly lower than those in the interleukin 1β group and the empty vector group(P＜0.01,P＜0.001).(4)Western blot assay results showed that compared with the interleukin 1β group and the empty vector group,the expression of the pro-apoptotic factor Bax protein in the sh-circSEC24A group significantly decreased,and the expression of the anti-apoptotic factor Bcl-2 protein increased significantly(P＜0.05);apoptosis and related activating factors cleaved-casepase3 and cleaved-casepase8 protein expressions were both reduced(P＜0.05).(5)ELISA and western blot assay results showed that compared with the interleukin 1β group and the empty vector group,the sh-circSEC24A group had lower levels of matrix metalloproteinase 9 and matrix metalloproteinase 13 protein(P＜0.05).These findings indicated that the expression of circSEC24A was abnormally increased in human synovial fibroblasts induced by interleukin 1β.Knocking down circSEC24A expression could promote the proliferation of human synovial fibroblasts and inhibit apoptosis,inflammatory factor release,and extracellular matrix degradation,suggesting that circSEC24A may be an important intervention target for early osteoarthritis.

Objective To explore the role of ferroptosis in DIC through bioinformatics analysis of hub genes involved in ferroptosis in doxorubicin-induced cardiomyopathy(DIC),combined with in vitro experimental validation.Methods Divalent iron fluorescence staining confirms the occurrence of ferroptosis in myocardial cells of DIC.The GSE207737 dataset was retrieved from the Gene Expression Comprehensive Database(GEO)and intersected with the FerrDb database to identify ferroptosis-related genes.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses of the intersected genes and intersecting the genes obtained from LASSO regression analysis and SVM-SFR machine learning methods were used to obtain ferroptosis hub genes for DIC.Real-time PCR was used to validate H9C2 cells in the control and DIC model groups,and Western blotting was used to further validate those whose bioinformatics and real-time PCR results that did not match.Results Thirty-eight ferroptosis-related genes in DIC were identified,and GO and KEGG analyses showed that these genes mainly participate in cell metabolism.Five hub genes for ferroptosis in DIC were obtained using machine learning methods:Mpc1,Prdx1,Kdm4a,Alox 12b,and Tfrc.Through in vitro experiments,the mRNA expression levels of Mpc1,Prdx1,and Kdm4a were downregulated in the DIC model group compared to those in the control group(P＜0.001),whereas the mRNA expression level of Alox12b was upregulated(P＜0.001).There were no significant differences in the mRNA or protein expression levels of Tfrc(P＞0.05).Conclusion Mpc1,Prdx1,Kdm4a,and Alox12b are key genes involved in ferroptosis in doxorubicin-induced cardiomyopathy and potential targets for the prevention and treatment of doxorubicin-induced cardiomyopathy in ferroptosis.

The incidence of age-related macular degeneration(AMD)is rising with the intensifying aging trend,be-coming a critical challenge that demands urgent solutions.Dry AMD(dAMD)accounts for approximately 80％of all AMD cases,and there is currently a lack of highly effective treatment options.In recent years,traditional Chinese medicine(TCM)has been proven to effectively treat dAMD through actions such as antioxidation,anti-apoptosis,anti-inflammation,and lipid-lowering.By reviewing domestic and international literature,this article discusses TCM monomers,extracts,and compound formulations for dAMD,analyzing their various mechanisms.Based on traditional TCM efficacy theories and in-tegrated with modern mechanisms of action,it targets the active components of TCM to elucidate the connection between effective medicinal targets of TCM and dAMD,thereby clarifying the efficacy and scientific basis of TCM monomers,com-pounds,and extracts in treating dAMD.The aim is to provide new perspectives for the prevention and clinical treatment of dAMD.