[摘 要] 目的：探讨lncRNA核内富集丰富转录本1（NEAT1）调控miR-1287-5p/DNA损伤诱导转录本4（DDIT4）轴对卵巢癌SKOV3细胞增殖、凋亡和侵袭的影响及其机制。方法：收集2023年6月至2024年6月期间湖北医药学院附属随州医院手术切除的27例卵巢癌患者的癌及癌旁组织标本，以及正常人卵巢上皮细胞IOSE80和卵巢癌细胞系SKOV3、CAOV3和A2780，RT-qPCR检测卵巢癌组织与细胞中lncRNA NEAT1、miR-1287-5p及DDIT4 mRNA表达。将SKOV3细胞分为Ctrl组、si-NC组、si-NEAT1组、si-NEAT1 + anti-miR-NC组、si-NEAT1 + anti-miR-1287-5p组、si-NEAT1 + vector组、si-NEAT1 + OE-DDIT4组。CCK-8法、克隆形成实验、流式细胞术、Transwell实验分别检测各组细胞增殖、凋亡及侵袭能力，WB法检测细胞中DDIT4、细胞周期蛋白D1（cyclin D1）、p53、迁移侵袭增强子1（MIEN1）蛋白水平。双萤光素酶报告基因验证lncRNA NEAT1与miR-1287-5p/DDIT4靶向结合关系，RNA pull-down实验、RNA免疫沉淀实验分别验证lncRNA NEAT1与miR-1287-5p、miR-1287-5p与DDIT4的靶向结合关系。另设pcDNA组（转染空载体pcDNA3.1）和pc-NEAT1 组（转染pcDNA-NEAT1）以验证NEAT1过表达效应。结果：卵巢癌组织中lncRNA NEAT1、DDIT4 mRNA表达水平显著高于癌旁组织（P < 0.05），而miR-1287-5p表达显著低于癌旁组织（P < 0.05）。敲低lncRNA NEAT1后，与Ctrl组和si-NC组相比，si-NEAT1组lncRNA NEAT1表达、DDIT4 mRNA表达均显著降低（均P < 0.05），miR-1287-5p 表达显著升高（P < 0.05）；而过表达lncRNA NEAT1则下调miR-1287-5p表达并上调DDIT4表达，呈现与敲低实验相反的调控效应；敲低lncRNA NEAT1后，克隆形成数、细胞增殖活性、细胞侵袭数均显著降低（均P < 0.05），细胞凋亡率显著升高（P < 0.05）；DDIT4、cyclin D1、MIEN1蛋白表达均显著降低（均P < 0.05），p53蛋白表达显著升高（P < 0.05）。进一步实验证实，anti-miR-1287-5p或OE-DDIT4均可减弱si-NEAT1对SKOV3细胞增殖和侵袭的抑制作用，同时减弱其对细胞凋亡的促进作用。lncRNA NEAT1靶向调控miR-1287-5p/DDIT4。结论：lncRNA NEAT1通过靶向调控miR-1287-5p/DDIT4轴促进SKOV3细胞增殖和侵袭，抑制细胞凋亡。

Obesity usually exacerbates the immunosuppressive tumor microenvironment (ITME), hindering CD8⁺ T cell infiltration and function, which further represents a significant barrier to the efficacy of immunotherapy. Herein, a multifunctional liposomal system (CR-Lip) for encapsulating celastrol (CEL) was utilized to remodel obesity-related ITME and improve cancer immunotherapy, wherein Ginsenoside Rg3 (Rg3) was detected interspersed in the phospholipid bilayer and its glycosyl exposed on the surface of the liposome. CR-Lip had a relatively uniform size (116.5 nm), facilitating favorable tumor tissue accumulation through the interaction between Rg3 and glucose transporter 1 overexpressed in obese tumor cells. Upon reaching the tumor region, CR-Lip was found to induce the immunogenic cell death (ICD) of HFD tumor cells. Notably, the level of PHD3 in HFD tumor cells was effectively boosted by CR-Lip to effectively block metabolic reprogramming and increase the availability of major free fatty acids fuel sources. In vivo, experiments studies revealed that the easy-obtained nano platform stimulated enhanced the production of various cytokines in tumor tissues, DC maturation, CD8⁺ T-cell infiltration, and synergistic anticancer therapeutic potency with aPD-1 (tumor inhibition rate = 82.1%) towards obesity-related melanoma. Consequently, this study presented an efficacious approach to tumor immunotherapy in obese mice by encompassing tumor eradication, inducing ICD, and reprogramming metabolism. Furthermore, it offered a unique insight into a valuable attempt at the immunotherapy of obesity-associated related tumors.

The interest in covalent drugs has resurged in recent decades, spurring the development of numerous specialized computational docking tools to facilitate covalent ligand design and screening. Herein, we present CarsiDock-Cov, a new paradigm distinguishing itself as the first deep learning (DL)-guided approach for covalent docking. CarsiDock-Cov retains the core components of its non-covalent predecessor, leveraging a DL model pretrained on millions of docking complexes to predict protein-ligand distance matrices, along with a dedicated-designed geometric optimization procedure to convert these distances into refined binding poses. Additionally, it incorporates several key enhancements specifically tailored to optimize the protocol for covalent docking applications. Our approach has been extensively validated on multiple public datasets regarding the docking and screening of covalent ligands, and the results indicate that our approach not only achieves comparably improved applicability compared to its non-covalent predecessor, but also exhibits competitive performance against various state-of-the-art covalent docking tools. Collectively, our approach represents a significant advance in covalent docking methodology, offering an automated and efficient solution that shows considerable promise for accelerating covalent drug discovery and design.

Objective To explore the predictive value of ultra-sound(US)combined with multi-sequence MRI in a clinical-deep learning radiomics nomogram(DLRN)for the short-term efficacy of neoadjuvant chemotherapy(NAC)in patients with triple-negative breast cancer(TNBC).Methods A total of 122 TNBC patients from five hospitals were retrospectively analyzed,and divided into training group(72 cases)and validation group(50 cases).The clinical and pathological data,NAC regimens,and imaging data were collected.The lesions and its surrounding 10-unit voxels from US and MRI images were retained as the region of interest(ROI).Pyradiomics software and a ResNet152 convolutional neural network(CNN)framework were used to extract radiomics and deep learning features to construct a clinical-DLRN with outcome dimension fusion.The receiver operating characteristic(ROC)curve and calibration curve were plotted,and five-fold cross-validation decision curve were used to verify the model's clinical effec-tiveness.Results The clinical-DLRN constructed by US combined with multi-sequence MRI showed that area under the curve(AUC)was 0.967[95％confidence interval(CI)0.782-0.967]and accuracy(ACC)was 0.900,respectively.The five-fold cross-validation decision curve showed good generalization,with the highest clinical net benefit between risk thresholds of 0.72 and 0.96.Conclusion The clinical-DLRN integrating US and multi-sequence MRI has the best efficacy in predicting the short-term efficacy of NAC in TNBC patients,which offering potential guidance for personalized TNBC treatment.

Objective To explore the predictive value of ultra-sound(US)combined with multi-sequence MRI in a clinical-deep learning radiomics nomogram(DLRN)for the short-term efficacy of neoadjuvant chemotherapy(NAC)in patients with triple-negative breast cancer(TNBC).Methods A total of 122 TNBC patients from five hospitals were retrospectively analyzed,and divided into training group(72 cases)and validation group(50 cases).The clinical and pathological data,NAC regimens,and imaging data were collected.The lesions and its surrounding 10-unit voxels from US and MRI images were retained as the region of interest(ROI).Pyradiomics software and a ResNet152 convolutional neural network(CNN)framework were used to extract radiomics and deep learning features to construct a clinical-DLRN with outcome dimension fusion.The receiver operating characteristic(ROC)curve and calibration curve were plotted,and five-fold cross-validation decision curve were used to verify the model's clinical effec-tiveness.Results The clinical-DLRN constructed by US combined with multi-sequence MRI showed that area under the curve(AUC)was 0.967[95％confidence interval(CI)0.782-0.967]and accuracy(ACC)was 0.900,respectively.The five-fold cross-validation decision curve showed good generalization,with the highest clinical net benefit between risk thresholds of 0.72 and 0.96.Conclusion The clinical-DLRN integrating US and multi-sequence MRI has the best efficacy in predicting the short-term efficacy of NAC in TNBC patients,which offering potential guidance for personalized TNBC treatment.

OBJECTIVE To know about the working mode and human resource status of pharmacy intravenous admixture services （PIVAS） in national medical institutions. METHODS Through questionnaire survey， the national PIVAS was invited to fill out questionnaire and statistical analysis was performed on the effective sample data related to PIVAS working mode and human resources in the questionnaire. RESULTS In this study， 761 PIVAS from 722 medical institutions of 29 provinces were involved in the questionnaire survey， with 471 valid questionnaires for working mode and 441 valid questionnaires for human resources survey. In terms of working mode， among 471 PIVAS， 292 PIVAS （62.0%） were in pharmacist-alone mode， and 176 PIVAS （37.4%） were in pharmacist-nurse cooperative mode； there was no significant difference in the types of medical orders received by PIVAS between these two working modes except for the other medical orders （P＞0.05）. In terms of human resource setting， among 441 PIVAS， the average number of total staff of single PIVAS was 24（16，33）， including 11（6，19） pharmacists， 7（2，13） nursing staff， and 3（1，5） workers； there was a statistically significant difference in the number of personnel among three groups （P＜ 0.01）. The per capita income of PIVAS respondents in 2019 was [7.9（4.8，10.7）]×104 yuan， and in 2021 it was [8.8（5.8，11.7）]× 104 yuan， with an increase of 9.0% compared to 2019. The difference between the two groups was statistically significant （P＜ 0.01）. CONCLUSIONS Medical institutions’ PIVAS in China had not fully implemented the pharmacist-alone work model， and some medical institutions had chosen a pharmacist-nurse cooperative mode. It is suggested that relevant departments formulate corresponding qualification requirements and training standards for nursing personnel as soon as possible based on sufficient research on PIVAS’s demand for nursing professionals.

OBJECTIVE To know about the pharmacy intravenous admixture charge and operation balance of pharmacy intravenous admixture services （PIVAS） in national medical institutions. METHODS Using questionnaire survey method， the national PIVAS leaders were invited to fill in the questionnaire， investigation and statistical analysis of the drug dispensing charge standard and the income and expenditure situations of PIVAS nationwide were conducted. RESULTS A total of 761 PIVAS completed the questionnaire， among which 466 PIVAS （61.2%） had already started implementing pharmacy intravenous admixture charge， mainly in tertiary hospitals. The charge standards for chemotherapy drugs and parenteral nutrition solutions were relatively high， while the standards for packaged drugs were the lowest， with differences in charge standards among provinces（P＜0.05）. Among the 25 provinces that reported annual drug preparation fee revenue， Hubei had the highest revenue in both 2019 and 2021. In 2019， the number of PIVAS with a balance of payments was more than that of PIVAS with an imbalance of payments， but the number of PIVAS with an imbalance of payments in 2021 exceeded the number of PIVAS with a balance of payments （P＜0.05）； among them， eight provinces were unbalanced in 2019 and 2021， such as Tianjin， Chongqing， Guizhou， etc. CONCLUSIONS PIVAS charge standards of the surveyed medical institutions in all provinces are not unified. It is suggested to improve the charge standard further， formulate the charge adjustment cycle， and promote a sustainable development of PIVAS.

Objective To explore the therapeutic mechanism of compound Yuye Decoction against diabetic cardiomyopathy(DCM).Methods Drugbank,Gene Cards,OMIM and PharmGKb databases were used to obtain DCM-related targets,and the core targets were identified and functionally annotated by protein-protein interaction network analysis followed by GO and KEGG enrichment analysis.The"Traditional Chinese Medicine-Key Component-Key Target-Key Pathway"network was constructed using Cytoscape 3.9.1,and molecular docking was carried out for the key components and the core targets.In the animal experiment,Wistar rat models of DCM were treated with normal saline or Yuye Decoction by gavage at low(0.29 g/kg)and high(1.15 g/kg)doses for 8 weeks,and the changes in cardiac electrophysiology and histopathology were evaluated.The changes in serum levels of LDH,CK,and CK-MB were examined,and myocardial expressions of PI3K,P-PI3K,Akt,P-AKT,BAX,IL-6,and TNF-α were detected using Western blotting.Results We identified 61 active compounds in Yuye Decoction with 1057 targets,3682 DCM-related disease targets,and 551 common targets between them.Enrichment of the core targets suggested that apoptosis,inflammation and the PI3K/Akt pathways were the key signaling pathways for DCM treatment.Molecular docking studies showed that the active components in Yuye Decoction including gold amidohydroxyethyl ester and kaempferol had strong binding activities with AKT1 and PIK3R1.In DCM rat models,treatment with Yuye Decoction significantly alleviated myocardial pathologies,reduced serum levels of LDH,CK and CK-MB,lowered myocardial expressions of BAX,IL-6 and TNF-α,and increased the expressions of P-PI3K and P-AKT.Conclusion The therapeutic effect of compound Yuye Decoction against DCM is mediated by its multiple active components that act on multiple targets and pathways to inhibit cardiomyocyte apoptosis and inflammatory response by regulating the PI3K/Akt signaling pathway.

Objective To explore the therapeutic mechanism of compound Yuye Decoction against diabetic cardiomyopathy(DCM).Methods Drugbank,Gene Cards,OMIM and PharmGKb databases were used to obtain DCM-related targets,and the core targets were identified and functionally annotated by protein-protein interaction network analysis followed by GO and KEGG enrichment analysis.The"Traditional Chinese Medicine-Key Component-Key Target-Key Pathway"network was constructed using Cytoscape 3.9.1,and molecular docking was carried out for the key components and the core targets.In the animal experiment,Wistar rat models of DCM were treated with normal saline or Yuye Decoction by gavage at low(0.29 g/kg)and high(1.15 g/kg)doses for 8 weeks,and the changes in cardiac electrophysiology and histopathology were evaluated.The changes in serum levels of LDH,CK,and CK-MB were examined,and myocardial expressions of PI3K,P-PI3K,Akt,P-AKT,BAX,IL-6,and TNF-α were detected using Western blotting.Results We identified 61 active compounds in Yuye Decoction with 1057 targets,3682 DCM-related disease targets,and 551 common targets between them.Enrichment of the core targets suggested that apoptosis,inflammation and the PI3K/Akt pathways were the key signaling pathways for DCM treatment.Molecular docking studies showed that the active components in Yuye Decoction including gold amidohydroxyethyl ester and kaempferol had strong binding activities with AKT1 and PIK3R1.In DCM rat models,treatment with Yuye Decoction significantly alleviated myocardial pathologies,reduced serum levels of LDH,CK and CK-MB,lowered myocardial expressions of BAX,IL-6 and TNF-α,and increased the expressions of P-PI3K and P-AKT.Conclusion The therapeutic effect of compound Yuye Decoction against DCM is mediated by its multiple active components that act on multiple targets and pathways to inhibit cardiomyocyte apoptosis and inflammatory response by regulating the PI3K/Akt signaling pathway.

Objective:To quantify the associations between periconceptional maternal homocysteine (HCY) and offspring′s birth weight and risk of small for gestational age (SGA) infant.Methods:The 19 984 mother-child pairs in this prospective cohort study were recruited from the Shanghai preconception cohort; the infants were delivered from 1 st September 2016 to 11 th November 2022. A standardized questionnaire was used to collect the mothers′ demographic information, medical history, dietary supplement use, and maternal complications during pregnancy, and their serum samples were collected. Serum HCY, folate, and vitamin B 12 were measured using chemiluminescent microparticle immunoassay based on serum sample drawn at enrollment. Birth weight data were obtained from medical records. Multiple imputation methods were applied to handle missing data in key variables. Multivariable linear regression and Poisson regression models were used to analyze the relationship between maternal HCY concentration during the periconceptional period and the birth weight and SGA risk of the offspring. Results:A total of 9 452 pairs were enrolled preconceptionally and the remaining 10 532 pairs were enrolled in early pregnancy. The proportion of mothers whose pregnancy age was greater than 35 years was 9.2% (1 832/19 984), the proportion of primiparous women was 76.5% (15 283/19 984), the proportion of pre-pregnancy overweight and obesity was 14.0% (2 804/19 984), the proportion of using folic acid supplements before pregnancy was 21.4% (4 272/19 984), and the proportion of those who supplemented with folic acid during early pregnancy was 85.2% (8 976/10 532); gestational diabetes mellitus was in 6.2% (1 245/19 984), gestational hypertensive syndrome in 3.6% (711/19 984). The birth weight of the offspring was (3 297±468) g, and there were 1 962 SGA children (9.8%). The HCY concentration in the overall population in appropriate for gestational age during the periconceptional period was (7.9±3.2) μmol/L, with (8.3±3.7) μmol/L in the preconception subgroup and (7.3±2.4) μmol/L in the early pregnancy subgroup. After adjustment for the covariates of perinatal demographic information, adverse pregnancy outcomes, serum folate and vitamin B 12, increased maternal periconceptional HCY was significantly associated with lower offspring birth weight ( β=-2.30, 95% CI -4.43--0.16, P=0.035). Only the early pregnancy subgroup was significantly associated with lower offspring birth weight ( β=-7.39, 95% CI-11.50--3.21, P<0.001). No association was found between peripregnancy HCY and offspring SGA risk. However, elevated HCY in early pregnancy was associated with an increased risk of SGA in the offspring ( RR=1.05, 95% CI 1.01-1.08, P=0.002). Periconceptional vitamin B 12 was a mediator of the association between HCY and offspring birth weight, accounting for 16.5%, 41.2% and 5.4% of its total effect in the overall periconceptional population, the pre-pregnancy subgroup and the early pregnancy subgroup, respectively. Conclusions:Maternal periconceptional HCY level is associated with lower birth weight in offspring, but not with the risk of SGA. Elevated maternal HCY in early pregnancy subgroup may be associated with increased risk of SGA in offspring.