BACKGROUND:Although breast cancer is a significant heterogeneous disease with an increasing global prevalence, precise prognostic evaluation is a vital aspect of designing personalized therapy strategies and upholding patients' survival rates. With the incorporation of artificial intelligence technology, in particular, machine learning, cancer prognosis and prediction have significantly been redefined. METHODS:In this study, we adopted a ten-fold cross-validation method to construct a Machine Learning-Derived Transcription Factor Signature (MDTS) across 108 algorithmic combinations. The optimal model was selected based on the highest average C-index across ten cohorts. We integrated single-cell data with multi-omics analysis to comprehensively assess the robustness of the MDTS model at both molecular and genomic levels. The MDTS demonstrated superior predictive power, outperforming 103 existing signatures and accurately predicting breast cancer outcomes across 10 independent cohorts. RESULTS:Our findings revealed that patients with low MDTS scores are more likely to benefit from immunotherapy, while the PAC-1 drug was identified as the most targeted agents to the chemotherapy with high MDTS score. CONCLUSIONS:These insights will open the door to delivering cutting-edge MDTS strategies to customizing breast cancer therapies.

Clear cell renal cell carcinoma (ccRCC) represents a major subtype of kidney cancer with variable prognosis. A comprehensive understanding of sex hormone-related pathways could potentially refine the prediction of patient outcomes in ccRCC. Patients from TCGA-KIRC (n = 528) and GSE22541 (n = 40) cohorts were analyzed. Sex-hormone-associated pathways were manually collected and calculated with the activated score, then subtypes were identified. Differential gene expression, pathway enrichment, and tumor-infiltrating immunocytes were assessed. A prognostic signature was developed using Cox analysis and LASSO regression. Immunohistochemistry (IHC) was performed to validate the protein level of key model gene in ccRCC tissues. Three distinct subtypes (C1, C2, C3) based on sex hormone pathway activation were discovered. C1 showed the most favorable prognosis (P = 0.00029). 1,094 genes were upregulated in C1 and 197 in C3. 20 risk-associated and 172 protective genes for ccRCC prognosis were identified. LASSO regression narrowed down to 33 genes for the sex-hormone-related-gene (SHAG) prognostic model. In the TCGA-KIRC cohort, the high-SHAG score group had a worse prognosis with an HR of 3.26 (95% CI: 2.334-4.555, P < 0.001). Validation in the GSE22541 cohort corroborated these findings. The nomogram incorporating the SHAG model demonstrated robust predictive accuracy higher than 0.75. IHC validation confirmed that ARHGEF17 protein levels were higher in early-stage ccRCC (stage I-II) compared to advanced-stage (stage III) tumors, supporting its prognostic relevance. The SHAG signature serves as a promising prognostic tool for ccRCC, providing insights into the role of sex hormone-related pathways in tumor progression. Further experimental and clinical validation is warranted to explore its potential in personalized therapy.

BackgroundBreast cancer is a heterogeneous malignancy with complex molecular characteristics, making accurate prognostication and treatment stratification particularly challenging. Emerging evidence suggests that lactylation, a novel post-translational modification, plays a crucial role in tumor progression and immune modulation.MethodsTo address breast cancer heterogeneity, we developed a machine learning-derived lactylation signature (MLLS) using lactylation-related genes selected through random survival forest (RSF) and univariate Cox regression analyses. A total of 108 algorithmic combinations were applied across multiple datasets to construct and validate the model. Immune microenvironment characteristics were analyzed using multiple immune infiltration algorithms. Computational drug-repurposing analyses were conducted to identify potential therapeutic agents for high-risk patients.ResultsThe MLLS effectively stratified patients into low- and high-risk groups with significantly different prognoses. The model demonstrated robust predictive power across multiple cohorts. Immune infiltration analysis revealed that the low-risk group exhibited higher levels of immune checkpoints (e.g., PD-1, PD-L1) and greater infiltration of B cells, CD4+ T cells, and CD8+ T cells, suggesting better responsiveness to immunotherapy. In contrast, the high-risk group showed immune suppression features associated with poor prognosis. Methotrexate was computationally predicted as a potential therapeutic candidate for high-risk patients, although experimental validation remains necessary.ConclusionThe MLLS represents a promising prognostic biomarker and may support personalized treatment strategies in breast cancer, particularly for identifying candidates who may benefit from immunotherapy.

Metabolic plasticity and ferroptosis are essential for colorectal cancer (CRC) progression. The effects and prognostic value of metabolic plasticity- and ferroptosis-related genes (MPFRGs) in CRC remain unclear. We established a prognostic model for CRC patients by identifying important genes in metabolic plasticity and ferroptosis. Data of CRC patients were retrieved from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus; MPFRG data were obtained from GeneCards and FerrDb. We performed functional (to explore differences between the two metabolic subtypes) and single-sample gene set (to assess the immune environment) enrichment analyses. Immunophenotype, tumor immunological dysfunction, and exclusion scores were assessed to determine patient immune responses. A least absolute shrinkage and selection operator-Cox regression model comprising 10 significant differentially expressed genes of metabolic plasticity and ferroptosis (MPFDEGs) was constructed using TCGA training cohort and validated using the GSE17536 and GSE39582 datasets. We established a nomogram comprising metabolic plasticity- and ferroptosis-based signatures, revealing the clinical application and potential molecular mechanisms underlying the role of MPFRGs in CRC. Our model (developed based on 10 MPFDEGs) is efficient for calculating the overall survival of CRC patients. Our findings provide new strategies for the clinical management and individualized treatment of these patients.

Acute lung injury (ALI) is a severe clinical respiratory condition characterized by high rates of mortality and morbidity, for which effective treatments are currently lacking. In this study, lipopolysaccharide (LPS) was used to induce ALI mice, demonstrating the efficacy of tetramethylpyrazine (TMP) in ameliorating ALI. Subsequent we perfored high-throughput sequencing analysis and used Targetscan 8.0 and miRWalk 3.0 databases to predict the interaction between microRNAs and destrin (DSTN), ultimately identifying miR-369-3p as the focus of the investigation. The adenovirus carrying miR-369-3p was administered one week prior to LPS-induced in order to assess its potential efficacy in ameliorating ALI in mice. The findings indicated that the overexpression of miR-369-3p resulted in enhanced lung function, reduced pulmonary edema, inflammation, and permeability in LPS-induced ALI mice, while the suppression of miR-369-3p exacerbated the damage in these mice. Furthermore, the beneficial effects of TMP on LPS-induced ALI were negated by the downregulation of miR-369-3p. The results of our study demonstrate that TMP mitigates LPS-induced ALI through upregulation of miR-369-3p. Consequently, the findings of this study advocate for the clinical utilization of TMP in ALI treatment, with miR-369-3p emerging as a promising target for future ALI interventions.

Background:The respiratory system is intensely damaged by acute lung injury (ALI). The anti-inflammatory effects of tetramethylpyrazine (TMP) against ALI have been confirmed, but it exhibits a short half-life. miR-194-5p could directly target Rac1, but the internalization rate of miRNA cells was low.Purpose:To explore the potential of the soft mesoporous organic silica nanoplatform (NPs) as carriers for delivery of TMP and miR-194-5p through the tail vein.Methods:NPs@TMP and NPs@PEI@miR-194-5p were added to the HUVEC cell-lines, in vitro, to observe the cell uptake and cytotoxic effects. In vivo experiments were conducted by injecting fluorescently labeled NPs through the tail vein and tracking distribution. Therapeutic and toxic side-effects were analyzed systemically.Results:In vitro study exhibited that NPs have no toxic effect on HUVECs within the experimental parameters and have excellent cellular uptake. The IVIS Spectrum Imaging System shows that NPs accumulate mainly in the lungs. NPs@TMP treatment can improved oxidative stress and inflammation levels in ALI mice and inhibited the TLR4/NLRP3/caspase 1 pathway. NPs@PEI@miR-194-5p can inhibit the Rac1/ZO-1/occludin pathway and improved endothelial cell permeability in ALI mice. The co-treatment of NPs@TMP and NPs@PEI@miR-194-5p can significantly improved the survival rates of the mice, reduced pulmonary capillary permeability and improved pathological injury in ALI mice.Innovation:This study combined traditional Chinese medicine, bioinformatics, cellular molecular biology and nanobiomedicine to study the pathogenesis and treatment of ALI. The rate of cellular internalization was improved by changing the shape and hardness of nanoparticles. NPs@TMP and NPs@PEI@miR-194-5p combined application can significantly improve the survival condition and pathological injury of mice.Conclusion:NPs loaded with TMP and miR-194-5p showed a greater therapeutic effect in ALI mice.

Acute lung injury (ALI) is a respiratory disorder characterized by severe inflammation of the alveoli and lung parenchyma. Tetramethylpyrazine (TMP), the main active compound in Ligusticum chuanxiong Hort (LC), can protect against lipopolysaccharide (LPS)-induced ALI. Our study aimed to investigate how TMP protects the endothelial cell barrier in pulmonary capillaries. We administered TMP intraperitoneally at different doses and found that acute lung injury in mice was improved, but not in a dose-dependent manner. TMP toxicity was tested in vitro. We observed that LPS-induced cytoskeletal remodeling was inhibited by TMP. Murine ALI was induced as follows: For the 1st hit, LPS (2 mg/kg) was injected intraperitoneally; after 16 h, for the 2nd hit, LPS (4 mg/kg) was instilled intratracheally. The mice in treatment groups had TMP or dexamethasone administered intraperitoneally 30 min prior to the 1st hit and 30 min past the 2nd hit. Mice were euthanized 24 h after the last injecting. We measured protein and mRNA levels using enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase real-time PCR (RT-qPCR), respectively. The ultrastructural analysis was performed with transmission electron microscopy (TEM) and the cytoskeleton was observed by immunofluorescence. Immunohistochemistry and Western blotting were used to detect protein expression in the Rac1/LIMK1/ZO-1/occludin signal pathway. The results showed that TMP treatment decreased inflammatory cell infiltration and alleviated LPS-induced damage in lung tissue. Also, TMP significantly inhibited the Rac1/LIMK1/ZO-1/occludin signaling pathway. Our findings show that using TMP during sepsis can protect the pulmonary microvascular endothelial cell barrier and suppress inflammation. Therefore, TMP may have a promising therapeutic role in preventing acute lung injury from sepsis.

Breast cancer is characterized by some types of heterogeneity, high aggressive behaviour, and low immunotherapeutic efficiency. Detailed immune stratification is a prerequisite for interpreting resistance to treatment and escape from immune control. Hence, the immune landscape of breast cancer needs further understanding. We systematically clustered breast cancer into six immune subtypes based on the mRNA expression patterns of immune signatures and comprehensively depicted their characteristics. The immunotherapeutic benefit score (ITBscore) was validated to be a superior predictor of the response to immunotherapy in cohorts from various datasets. Six distinct immune subtypes related to divergences in biological functions, signatures of immune or stromal cells, extent of the adaptive immune response, genomic events, and clinical prognostication were identified. These six subtypes were characterized as immunologically quiet, chemokine dominant, lymphocyte depleted, wounding dominant, innate immune dominant, and IFN-γ dominant and exhibited features of the tumor microenvironment (TME). The high ITBscore subgroup, characterized by a high proportion of M1 macrophages:M2 macrophages, an activated inflammatory response, and increased mutational burden (such as mutations in TP53, CDH1 and CENPE), indicated better immunotherapeutic benefits. A low proportion of tumor-infiltrating lymphocytes (TILs) and an inadequate response to immune treatment were associated with the low ITBscore subgroup, which was also associated with poor survival. Analyses of four cohorts treated with immune checkpoint inhibitors (ICIs) suggested that patients with a high ITBscore received significant therapeutic advantages and clinical benefits. Our work may facilitate the understanding of immune phenotypes in shaping different TME landscapes and guide precision immuno-oncology and immunotherapy strategies.

目的 观察川芎嗪对脂多糖诱导的小鼠ALI血管通透性的保护作用并探讨其可能分子机制.方法 C57BL/6 小鼠(n =50)随机分为5 组:CON组(n =10)、ALI组(n =10)、TMP低(50 mg/kg,n =10),中(100 mg/kg,n =10),高(150 mg/kg,n =10)浓度干预组.ELISA检测IL-1β及TNF-α水平;HE染色观察肺部病理变化;湿干比检测肺水肿程度;BALF白细胞计数、总蛋白量测定及EB实验检测肺血管通透性;WB检测P-mTOR、RhoA蛋白水平;电镜下观察内皮细胞间紧密连接.结果 ALI组IL-1β、TNF-α水平较于CON组升高(P均<0.05);中浓度TMP组IL-1β、TNF-α水平下降明显(P均<0.05),低浓度和高浓度TMP组无显著变化(P均>0.05).HE结果提示TMP中浓度可以显著减缓肺损伤的病理变化.ALI组湿干比高于CON组(P<0.01),TMP治疗可降低湿干比(P<0.05).TMP可降低BALF白细胞计数(P<0.01)、蛋白浓度(P<0.01)及Evans Blue渗漏(P<0.05).ALI组P-mTOR(P<0.01)及RhoA(P<0.01)蛋白表达量高于CON组.TMP组P-mTOR(P<0.01)及RhoA蛋白表达量下降(P<0.05).ALI组紧密连接受损,TMP治疗后紧密连接断裂减轻.结论 TMP可通过活化mTOR/RhoA通路改善肺血管通透性从而对LPS引起的小鼠ALI起到保护作用.

目的:探讨川芎嗪(TMP)对脂多糖(LPS)诱导的小鼠急性肺损伤(ALI)的预防作用及对Ras相关C3肉毒素底物1(Rac1)/LIM激酶1(LIMK1)信号通路的影响.方法:将C57BL/6小鼠随机分为6组:对照组、模型组(LPS组)、地塞米松(Dex)组及TMP(40、80和120 mg/kg)组.模型组小鼠第1次采用LPS腹腔注射,16 h后第2次采用气管滴注LPS诱导小鼠ALI模型;TMP组小鼠在LPS第1次腹腔注射前30 min及第2次气管滴注LPS后30 min时腹腔注射TMP;Dex组小鼠与TMP组同一时间腹腔注射3 mg/kg Dex.LPS气管滴注24 h后检测小鼠外周血氧饱和度(SpO2);取小鼠肺组织,计算肺组织湿干重比值(W/D);ELISA法测定肺组织中白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α);HE染色观察肺组织病理学变化;Western blot检测肺组织中Rac1和LIMK1蛋白水平;支气管肺泡灌洗液中检测白细胞含量及蛋白浓度.结果:与对照组比较,模型组小鼠SpO2降低(P<0.05),W/D升高(P<0.01),肺组织中IL-1β和TNF-α水平均升高(P<0.01);肺泡壁增厚,大量红细胞渗出,可见片状出血;支气管肺泡灌洗液中白细胞总数及蛋白浓度增高(P<0.01);Rac1和p-LIMK1蛋白表达水平显著升高(P<0.01).与模型组相比,Dex组和TMP组W/D降低(P<0.01),肺组织IL-1β和TNF-α水平降低(P<0.01),肺泡间质水肿及出血减轻,红细胞渗出减少;支气管肺泡灌洗液中白细胞总数及蛋白浓度降低(P<0.01).与模型组相比,Dex组及低、中剂量TMP组Rac1蛋白水平降低(P<0.01),p-LIMK1在Dex组及TMP组也显著降低(P<0.05,P<0.01),LIMK1在各组的表达无显著差异.结论:TMP可通过抑制Rac1/LIMK1信号通路降低毛细血管通透性,从而减轻LPS诱导的小鼠ALI.

该文旨在从Ras相关的C3肉毒素底物1（Ras-related C3 botulinum toxin substrate 1,Rac1）/LIM激酶1（LIM kinase 1,LIMK1）通路探讨川芎嗪（tetramethylpyrazine,TMP）对脂多糖（lipopolysaccharide,LPS）诱导的人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVECs）骨架重构的保护作用。采用Ⅷ因子免疫荧光鉴定原代内皮细胞;分别用0、50、100、200、300 ng/mL的LPS处理对数生长期的原代HUVECs;采用低、中、高3个剂量的TMP对HUVECs预处理12 h;利用F-actin免疫荧光染色观察细胞骨架变化;通过跨膜电阻值（transepithelial electrical resistance,TEER）和FITC-葡聚糖Transwell实验检测人脐静脉内皮细胞通透性;Western blot检测Rac1、LIMK1的表达以及相应磷酸化水平;免疫荧光观察磷酸化LIMK1蛋白的细胞定位和表达水平变化;实时荧光定量PCR（quantitative real-time PCR,qPCR）检测Rac1和LIMK1的mRNA水平。结果显示,Ⅷ因子免疫荧光鉴定原代提取的内皮细胞符合要求;HUVECs经LPS诱导后,镜下形态由卵圆形向长梭形改变,纤维状肌动蛋白（filament actin,F-actin）增多增粗,TEER降低及单层内皮细胞通透性增加;Rac1、LIMK1的表达及磷酸化水平呈LPS梯度依赖性升高（P＜0.05）;低浓度的TMP降低LPS诱导的F-actin增多增粗作用,并降低Rac1、LIMK1的表达及其磷酸化水平（P＜0.05）;高浓度的TMP可以抑制Rac1激活（P＜0.05）,但对LPS诱导的HUVECs细胞骨架重构和通透性增加没有保护作用（P＞0.05）。该研究结果表明,TMP通过Rac1/LIMK1通路保护LPS诱导的HUVECs骨架重构,抑制由LPS诱导的HUVECs通透性增大,对临床运用TMP改善由内皮细胞高通透性引起的急性肺损伤提供理论依据。

目的 观察川芎嗪(Tetramethylpyrazine,TMP)对细菌脂多糖(LPS)诱导的脐静脉内皮细胞屏障损伤(HUVEC)的保护作用,并探索可能的作用机制.方法 将鉴定好的人脐带静脉内皮细胞根据加药处理因素不同,将实验分为空白对照组、LPS组,LPS+TMP组.对三组进行免疫荧光细胞骨架染色及细胞通透性实验、跨内皮电阻实验.应用Westernblot及qPCR方法进行mTOR表达量的定量分析.结果 免疫荧光细胞骨架染色表明与空白组相比,LPS组应力纤维明显增多、增粗.而应用TMP治疗后应力纤维变少、变细.跨内皮电阻实验表明,加药刺激前三组电阻值差无统计学意义(P＞0.05).加入药物刺激后,LPS组较空白组,电阻值明显降低,而TMP组较LPS组明显增加(P＜0.05).细胞通透性实验表明加药刺激后,LPS组较空白组,通透系数明显增加,而TMP组较LPS组明显降低(P＜0.05).Westernblot及qPCR方法表明LPS组较空白组mTOR表达量增加,而TMP治疗之后mTOR表达量较LPS组下降.结论 TMP可通过调控mTOR的表达量对LPS诱导的HUVEC内皮细胞屏障破坏起到保护作用.

TP53 is the most frequently mutated gene in lung adenocarcinoma (LUAD). The tumor immune microenvironment (TIM) is considered a vital factor that influences tumor progression and survival rate. The influence of TP53 mutation on TIM in LUAD has not been fully studied. Here we systematically investigated the relationship and potential mechanisms between TP53 mutation status and immune response in LUAD. We constructed an immune prognostic model (IPM) using immune associated genes, which were expressed differentially between the TP53 mutant and wild type LUAD patients. We discovered that TP53 mutations were significantly associated with 5 immune related biological processes. Thirty-six immune genes were expressed differentially between TP53 mutant and wild type LUAD patients. An IPM was constructed using 3 immune genes to differentiate the prognostic survival in LUAD. The high-risk LUAD group displayed significantly higher proportions of dendritic cell resting, T cell CD4 memory resting and mast cell resting, and significantly low proportions of dendritic cell activated, T cell CD4 memory activated, and mast cell activated. Moreover, IPM was found to be an independent clinical feature and can be used to predict immunotherapy responses. In summary, we constructed and validated an IPM using 3 immune related genes, which provides a better understanding of the mechanism from an immunological perspectives.

急性呼吸窘迫综合征(acute respiratory distress syndrome,ARDS)是一种ICU 中常见的病死率较高的临床综合征,ARDS发生时,富含蛋白质的液体在肺泡中积聚,阻止肺部充满足够的空气,从而导致到达血液中的氧气减少,发生低氧血症[1 -2].急性肺损伤(acute lung injury,ALI )由严重的感染、外伤、休克、吸入有害气体及中毒等直接或间接因素引起的全身炎症反应综合征在肺部的表现,以肺泡及肺实质发生急性炎症为主要病理特征,其特点是发生低氧血症、非心源性肺水肿、肺顺应性降低和广泛的毛细血管渗漏[3].

目的:探究过表达miR-122 a是否可提高肝癌细胞HepG2对化疗药物的敏感性,并阐明其提高化疗药物敏感性的部分机制.方法:构建过表达miR-122 a的HepG2细胞系(miR-122a肿瘤细胞组),并与HepG2细胞(肿瘤细胞组)置于含不同浓度顺铂的培养液中,CKK-8法观察miR-122a上调对HepG2细胞存活率的影响,Annexin V-PE/7-AAD双染法观察miR-122a上调对HepG2细胞凋亡率的影响;Western blot检测miR-122a肿瘤细胞组和肿瘤细胞组中bcl-2和p53蛋白的表达水平.结果:低铂浓度顺铂条件下,miR-122 a肿瘤细胞组细胞的存活率低于肿瘤细胞组细胞的存活率,miR-122 a肿瘤细胞组24 h细胞的凋亡率高于肿瘤细胞组(P<0.05);Western blot法检测bcl-2和p53蛋白结果显示,在低浓度顺铂溶液中,miR-122a肿瘤细胞组bcl-2表达量低于肿瘤细胞组(P<0.05),miR-122a肿瘤细胞组p53表达量高于肿瘤细胞组(P<0.05).结论:过表达miR-122a能上调p53蛋白表达、下调bcl-2蛋白表达,增加HepG2细胞对顺铂敏感性,为肝癌患者提供基因治疗和降低肿瘤细胞耐药提供了理论和实验基础.