[摘 要] 目的：探讨核仁小RNA（snoRNA）71A通过TLR3/PD-L1通路对食管鳞状细胞癌（ESCC）细胞增殖、迁移及侵袭能力的影响。方法：qPCR检测52例ESCC患者的配对肿瘤组织、癌旁组织，以及人ESCC细胞中SNORA71A的表达情况。将反义寡核苷酸（SNORA71A-ASO-29、SNORA71A-ASO-102和NC-ASO）或小干扰RNA（siTLR3、siTLR3-NC）分别转染至人ESCC TE-1细胞，分别记为SNORA71A-ASO1组、SNORA71A-ASO2组、NC-ASO组及siTLR3组、siNC组。另外将过表达质粒pcDNA3.1-SNORA71A和pcDNA3.1空载体质粒分别转染至TE-1细胞，分别记为SNORA71A组和Vector组。细胞功能学实验（MTS实验、划痕愈合实验，以及Transwell侵袭实验）评估各组细胞在敲低或过表达SNORA71A后增殖、迁移和侵袭能力的变化。高通量转录组测序筛选SNORA71A下游作用靶基因，基因本体论（GO）和京都基因与基因组百科全书（KEGG）功能富集分析预测SNORA71A下游作用靶基因可能参与的生物学过程和信号通路。qPCR检测测序筛选出的下游作用靶基因TLR3在ESCC组织以及细胞中的表达。同时，qPCR和WB检测TE-1细胞在敲低或过表达TLR3后PD-L1 mRNA和蛋白质表达变化。细胞功能学实验检测TLR3敲低对SNORA71A所促进的TE-1细胞恶性生物行为（增殖、迁移、侵袭）的影响。结果：在52例ESCC患者的肿瘤组织以及ESCC细胞中SNORA71A表达均呈高水平（P < 0.01或P < 0.05）。敲低SNORA71A抑制TE-1细胞增殖、迁移和侵袭（P < 0.01或P < 0.05），过表达SNORA71A则促进TE-1细胞增殖、迁移和侵袭（P < 0.01或P < 0.05）。高通量转录组测序筛选到TLR3为SNORA71A下游作用靶基因。TLR3在ESCC组织以及TE-1细胞中呈低表达（P < 0.01或P < 0.05）。TLR3正向调控PD-L1 mRNA和蛋白质表达（P < 0.01或P < 0.05）。细胞功能学实验中，TLR3可以部分削弱SNORA71A对PD-L1表达的调控（P < 0.01）。结论：SNORA71A通过调控TLR3/PD-L1通路调节TE-1细胞增殖、迁移和侵袭。

Background/Aims: Hepatocellular carcinoma (HCC) is characterized by profound transcriptomic dysregulation, yet the mechanism(s) by which DNA methylation is coordinated with chromatin modifications to regulate alternative splicing during tumorigenesis remains poorly understood. Methods: Using prospectively paired multi-omics data obtained from metabolic dysfunction-associated steatotic liver disease (MASLD)-HCC patients and coupled with a premalignant MASLD cohort, we have uncovered a previously unrecognized gene-regulatory axis centered on TACC3 isoform-switching. Results: In the non-tumoral context, the TACC3-201 isoform directly engages the histone acetyltransferase KAT2A to coordinate the regulation of NOTCH4 signaling. In HCC, this regulatory axis is disrupted whereby FOXM1 overrides DNMT1-mediated methylation, upregulating TACC3, and decoupling TACC3 from the KAT2A-associated NOTCH4 co-expression module. This rewiring is licensing tumor-specific cell-cycle progression and epigenetic plasticity. Thus, FOXM1 reshapes the TACC3-KAT2A interaction, while DNMT1 drives context-dependent DNA methylation, activating the CDK1-inhibitory kinase PKMYT1. Conclusions We uncovered TACC3-KAT2A as an emerging regulatory axis caused by alternative splicing in HCC and propose FOXM1-driven TACC3 inhibition to synergistically disrupt mitotic fidelity and transcriptional regulation, potentially offering new therapeutic avenues for HCC with reduced toxicity to the normal liver.

[摘 要] 目的：探讨麻疹-流行性腮腺炎-风疹联合减毒活疫苗（MMR）通过诱导免疫原性细胞死亡（ICD）对三阴性乳腺癌（TNBC）的免疫调节机制。方法：体外实验采用CCK-8法检测MMR对TNBC细胞4T1和MDA-MB-231存活率的影响，免疫荧光法检测MDA-MB-231细胞中钙网蛋白（CALR）表达，磷钼酸比色法检测MDA-MB-231细胞内ATP的含量，ELISA检测细胞上清液中高迁移率族蛋白B1（HMGB1）的表达水平，通过GEPIA网站分析CALR和HMGB1对乳腺癌患者生存期的影响。体内实验中经皮下注射4T1-Luc细胞于雌性BALB/c小鼠乳腺脂肪垫构建TNBC小鼠模型。设Control组、盐酸阿霉素（Dox）组、MMR组，按照分组每两天静脉给药1次，连续给药3次。动物活体成像系统观察肿瘤体积变化；记录小鼠体质量及各器官指数初步评估疫苗安全性；H-E染色观察肿瘤病理变化；免疫组化和Western blotting检测肿瘤中CALR和HMGB1蛋白的表达。流式细胞术分析小鼠脾T淋巴细胞亚群及肿瘤树突状细胞和巨噬细胞比例；免疫组化检测肿瘤CD8+ T淋巴细胞浸润；ELISA检测血清中促炎细胞因子IL-6和TNF-α含量。结果：MMR对4T1细胞的活性抑制作用较弱；MMR对MDA-MB-231细胞抑制率达90.39%（P < 0.001）。与Control组相比，MMR组MDA-MB-231细胞中ICD相关蛋白（CALR、ATP、HMGB1）表达上调（P < 0.001）。GEPIA网站分析表明，CALR和HMGB1高表达的患者总生存期有所延长。MMR延缓了荷瘤小鼠肿瘤体积增长（P < 0.001），延长小鼠的生存期，且28 d内MMR组荷瘤小鼠体重与Control组无显著性差异（P > 0.05）；H-E染色显示，与Control组相比，MMR组小鼠肿瘤组织坏死区域扩大且有免疫细胞浸润；MMR上调肿瘤ICD相关蛋白CALR、HMGB1的表达水平；MMR组脾CD4+、CD8+ T淋巴细胞及肿瘤树突状细胞和M1型巨噬细胞的比例增加（P < 0.05），CD8+ T淋巴细胞在肿瘤组织中的浸润增加（P < 0.001），且促炎细胞因子IL-6和TNF-α水平升高（P < 0.01）。结论：MMR减毒活疫苗对TNBC的抗肿瘤作用可能与诱导肿瘤发生ICD进而激活抗肿瘤免疫相关。

Poly(ADP-ribose) polymerase 1 (PARP1) is a multifunctional protein involved in diverse cellular functions, notably DNA damage repair. Pharmacological inhibition of PARP1 has therapeutic benefits for various pathologies. Despite the increased use of PARP inhibitors, challenges persist in achieving PARP1 selectivity and effective blood-brain barrier (BBB) penetration. The development of a PARP1-specific positron emission tomography (PET) radioligand is crucial for understanding disease biology and performing target occupancy studies, which may aid in the development of PARP1-specific inhibitors. In this study, we leverage the recently identified PARP1 inhibitor, AZD9574, to introduce the design and development of its ¹⁸F-isotopologue ([¹⁸F]AZD9574). Our comprehensive approach, encompassing pharmacological, cellular, autoradiographic, and in vivo PET imaging evaluations in non-human primates, demonstrates the capacity of [¹⁸F]AZD9574 to specifically bind to PARP1 and to successfully penetrate the BBB. These findings position [¹⁸F]AZD9574 as a viable molecular imaging tool, poised to facilitate the exploration of pathophysiological changes in PARP1 tissue abundance across various diseases.

Autoimmune diseases, cancers, and viral infections pose significant global health threats, characterized by chronic pathology, unregulated cellular proliferation, and rapid transmission, respectively, requiring urgent early warning and treatment strategies. Antibodies, primarily classified into autoantibodies and therapeutic antibodies based on their clinical roles, provide essential information and show considerable value in the precise diagnosis and treatment of these serious diseases. Among the technologies utilized in bioanalysis, electrochemical biosensors, with their unique advantages of rapid response, high sensitivity, miniaturization, cost-effectiveness and user-friendly operation, have been developed as a trending technology for precise diagnostic and therapeutic drug monitoring. This review systematically summarizes the relationships and roles of clinically relevant antibodies in autoimmune diseases, cancers, and viral infections, while detailing the composition, strategies, development, and application trends of relevant electrochemical biosensors. Furthermore, it highlights the remaining challenges and opportunities for the advancement and prospects of electrochemical sensors in the context of clinically relevant antibodies.

Background: As of 2019, according to the Health Indicators of the Mongolian population, diseases of the digestive system ranked second in terms of morbidity, with gallbladder, biliary tract, and pancreatic diseases accounting for 14.2%, and the proportion of surgeries related to these diseases is high. Aim: Explore the ingredients and recipes of traditional medicine used for gallbladder disease. Materials and Methods: “The four foundations of medicine-Анагаах ухааны дөрвөн үндэс” and their interpretations, and source works were selected, and hermeneutic methodology, comparative method of source studies, historical classification method, and analysis-synthesis method were used. Results: A total of 43 recipes for treating gallstone disease were selected from the source texts. In terms of the method of treatment, all of them were written as hot and cold. The medicine for gallstone heat disease was directly written in “Manag Renchinjunai-Mанаг рэнчинжунай”. The two recipes were explained as new and old in the “Four springs-Рашааны дусал”. Although the contents written in other source books are the same as “The four foundations of medicine-Анагаах ухааны дөрвөн үндэс”, the “The four foundations of medicine-Анагаах ухааны дөрвөн үндэс” are very clearly explained as medicines to be taken, decoctions, and medicines to be given when the effects of heat are delayed. There is no difference in the following books and sutras regarding treatment, food and drink, and events. The method of adding mantras to the national method of Uvdis is not written in other sutras. The medicine for gallstone cold disease was directly written in “Manag Renchinjunai-Mанаг рэнчинжунай”. The amount of medicine in the “Four springs-Рашааны дусал” was carefully calculated. “The four foundations of medicine-Анагаах ухааны дөрвөн үндэс”are astringent, emetic, laxative, stopping the tail of the tail, and cutting off the tail of the disease and replacing it. The method of treating the disease is a method of adding charms to the root, and the “Four springs-Рашааны дусал” are also explained. The food and drink and the event are not written in the Mанаг рэнчинжунай, and there is no difference in other books and scriptures. Conclusion While many recipes for treating gallbladder diseases have been discovered in traditional treatises, the number of recipes and medicines for treating gallbladder heat diseases is greater than the number of medicines for treating gallbladder cold diseases.

[摘 要] 目的：基于生物信息学分析和体内、体外实验研究长链非编码RNA00511（LINC00511）敲减对非小细胞肺癌细胞增殖、凋亡、侵袭等恶性生物学行为的影响，并初步探究其作用机制。方法：通过基因表达谱交互分析（GEPIA）数据库分析LINC00511在非小细胞肺癌的表达水平，及其与患者肿瘤分期、生存期等临床特征、肿瘤细胞恶性生物学行为有关基因的相关性；利用shRNA慢病毒载体构建LINC00511敲减的H1299肺癌细胞株，克隆形成实验、划痕愈合实验和流式细胞术分别检测对H1299细胞增殖、迁移、细胞周期和凋亡能力的影响，qRT-PCR检测相关调控基因表达，WB法检测肿瘤相关蛋白的表达；构建裸鼠皮下移植瘤模型，取瘤组织进行免疫组织化学实验检测Ki67表达情况。结果：GEPIA数据库分析表明LINC00511在非小细胞肺癌组织中表达水平升高，且与该病的临床分期情况相关（P < 0.05），LINC00511与肺癌中CASP3、CCNB1、CDK4等多种基因表达均有相关性（P < 0.01）；LINC00511敲减可抑制细胞的克隆形成和迁移能力、促进肺癌细胞凋亡并影响细胞周期进展（P < 0.05，P < 0.01）；LINC00511敲减可下调肺癌细胞CCNB、CDK4、TGF-β1基因的表达（P < 0.01），对CCND1、VEGFA基因表达无明显影响，LINC00511敲减可抑制细胞内MMP9、CTNNB1表达，上调CASP3的表达（P < 0.05，P < 0.01）；裸鼠体内实验证实，LINC00511敲减可抑制移植瘤体组织内Ki67的表达（P < 0.01）。结论：LINC00511在非小细胞肺癌组织中呈高表达，与肺癌临床分期和多种基因表达具有相关性，LINC00511敲减可能通过影响相关基因、蛋白的表达，抑制肺癌H1299细胞的恶性生物学行为。

[摘 要] 目的：探讨黏蛋白13（MUC13）在肺腺癌组织中的表达及其对A549细胞增殖、凋亡、迁移、侵袭及EMT的影响与可能的机制。方法：通过癌症基因组图谱（TCGA）和高通量基因表达（GEO）数据库分析MUC13在肺腺癌组织与正常肺组织、癌旁组织中的差异表达。qPCR法和WB法检测人肺腺癌细胞NCI-H1395、NCI-H1975、H1299、A549和人正常肺上皮细胞BEAS-2B中MUC13 mRNA和蛋白的表达水平。利用siRNA技术敲低A549细胞中MUC13表达，实验分为si-MUC13组、NC组和si-MUC13+IGF-1组。通过克隆形成实验、流式细胞术和Transwell实验分别检测敲低MUC13对A549细胞增殖、细胞周期、凋亡、迁移和侵袭的影响，WB法检测敲低MUC13对A549细胞上皮钙黏素（E-cadherin）、神经钙黏素（N-cadherin）、波形蛋白（vimentin）、EGFR、p-EGFR、PI3K、p-PI3K、AKT、p-AKT等蛋白表达的影响。结果：MUC13 mRNA和蛋白在肺腺癌组织和细胞中均呈高表达（均P<0.01），选取表达水平较高的A549细胞进行后续实验。敲低MUC13后，A549细胞的增殖能力显著降低，G0/G1期的细胞数量显著增多、G2/M期及S期的细胞数量显著减少，细胞凋亡率显著升高，细胞迁移及侵袭能力均显著降低（均P<0.01）；A549细胞中E-cadherin表达显著上调，N-cadherin、vimentin表达显著下调，p-EGFR/EGFR、p-PI3K/PI3K、p-AKT/AKT比值均显著降低（均P<0.01）；再加入IGF-1处理后，A549细胞中p-EGFR/EGFR、p-PI3K/PI3K、p-AKT/AKT比值均显著升高（均P<0.01）。结论：MUC13在肺腺癌组织和细胞中均呈高表达，其可能通过激活EGFR/PI3K/AKT信号通路促进A549细胞增殖、迁移、侵袭和EMT。

[摘 要] 目的：探究茯苓酸（PA）是否通过AKT/MDM2/p53通路影响结直肠癌HCT116细胞的恶性生物学行为。方法：常规培养HCT116细胞，并将其分为对照组、MK-2206（AKT抑制剂）组、PA低浓度（PA-L）组、PA高浓度（PA-H）组、PA-H+ SC79（AKT激活剂）组。CCK-8法、细胞克隆形成实验、流式细胞术、Transwell、qPCR法和WB法实验分别检测各组HCT116细胞的增殖活力，克隆形成能力，细胞凋亡，迁移、侵袭能力，E-cadherin、N-cadherin和vimentin mRNA表达以及AKT/MDM2/p53通路相关蛋白的表达。结果：PA可明显抑制HCT116细胞的增殖活力（P<0.05）、克隆形成能力（P<0.05）、迁移和侵袭能力（P<0.05），诱导其凋亡（P<0.05），抑制N-cadherin、vimentin mRNA的表达（P<0.05），促进E-cadherin mRNA的表达（P<0.05），抑制AKT、MDM2的磷酸化水平（P<0.05），促进p53蛋白的表达（P<0.05）；AKT抑制剂MK-2206可模拟PA的作用（均P<0.05），而其激活剂SC79则可逆转PA的作用（均P<0.05）。结论：PA通过调控AKT/MDM2/p53信号通路来抑制HCT116细胞的增殖、迁移和侵袭并诱导其凋亡。

[摘 要] 目的：开发针对结直肠癌（CRC）个性化治疗的新抗原肽疫苗，探讨新抗原肽及其诱导的新抗原反应性T（NRT）细胞治疗CRC的可行性和有效性。方法：提取小鼠结肠癌CT26细胞的DNA和RNA，采用全外显子和转录组测序分析肿瘤基因的突变及表达。通过基于机器学习的新抗原预测体系，筛选、合成具有高免疫原性多肽。用合成的多肽经皮下注射免疫小鼠，通过流式细胞术检测免疫鼠脾细胞的IFN-γ分泌水平，筛选具有强免疫原性多肽。用免疫原性多肽负载小鼠骨髓来源的树突状细胞（BMDC）免疫结肠癌建模小鼠，通过ELISPOT检测效应细胞分泌IFN-γ的能力，时间分辨荧光免疫分析法检测免疫鼠脾细胞对相应靶细胞的杀伤力，观察荷瘤小鼠肿瘤生长情况和小鼠存活期。结果：新抗原肽Glud1-V546I具有更强的诱导NRT细胞分泌IFN-γ的能力（P < 0.000 1）。与野生肽（Glud1-WT）相比，Glud1-V546I在荷瘤鼠体内诱导的NRT细胞有更高的IFN-γ分泌能力（P <0.000 1）和细胞毒作用（P < 0.000 1）。同时，Glud1-V546I能明显抑制小鼠肿瘤生长（P < 0.001）并延长荷瘤鼠的生存期（P < 0.01）。结论：小鼠CT26细胞的新抗原肽Glud1-V546I能够显著促进小鼠NRT细胞的IFN-γ的分泌，用其制备的DC疫苗在结肠癌荷瘤鼠体内显示出有效的抗肿瘤反应，提示开发基于新抗原的CRC个性化免疫治疗是可能的。