BackgroundDrug-induced liver injury (DILI) is a leading cause of drug development failures during clinical trials and post-market introduction. Current biomarkers, such as ALT and AST, lack the necessary specificity and sensitivity needed for accurate detection. Exosomes, which protect LncRNAs from RNase degradation, could provide reliable and easily accessible options for biomarkers.Materials and methodsRNA-sequencing was used to identify differentially expressed LncRNAs (DE-LncRNAs), followed by isolation of LncRNAs from plasma exosomes in this study. Exosome characterization was conducted by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blot (WB). Bioinformatics analysis included functional enrichment and co-expression network analysis. Five rat models were established, and quantitative real-time PCR was used to verify the specificity and sensitivity of two candidate exosomal LncRNAs.ResultsThe APAP-induced hepatocellular injury model was successfully established for RNA-sequencing, leading to the identification of several differentially expressed exosomal LncRNAs. Eight upregulated exosomal DE-LncRNAs were selected for validation. Among them, NONRATT018001.2 (p < 0.05) and MSTRG.73954.4 (p < 0.05) exhibited a more than 2-fold increase in expression levels. In hepatocellular injury and intrahepatic cholestasis models, both NONRATT018001.2 and MSTRG.73954.4 showed earlier increases compared to serum biomarkers ALT and AST. However, no histological changes were observed until the final time point. In the fatty liver model, NONRATT018001.2 and MSTRG.73954.4 increased earlier than ALT and AST at 21 days. By the 7th day, minor steatosis was evident in liver tissue, while the expression levels of the two candidate exosomal LncRNAs exceeded 2 and 4 times, respectively. In the hepatic fibrosis model, NONRATT018001.2 and MSTRG.73954.4 showed increases at every time point. By the 49th day, hepatocellular necrosis and fibrosis were observed in the liver tissue, with NONRATT018001.2 showing an increase of more than 8 times. The specificity of the identified exosomal DE-LncRNAs was verified using a myocardial injury model and they showed no significant differences between the case and control groups.ConclusionNONRATT018001.2 and MSTRG.73954.4 hold potential as biomarkers for distinguishing different types of organ injury induced by drugs, particularly enabling early prediction of liver injury. Further experiments, such as siRNA interference or gene knockout, are warranted to explore the underlying mechanisms of these LncRNAs.

Legubicin, a novel prodrug based on doxorubicin, has both albumin-binding and legumain-activating properties. The aim of this study was to develop and validate a UHPLC-MS/MS method for investigating the in vivo pharmacokinetics and tissue distribution profiles of legubicin in rats and tumor-bearing mice following intravenous administration, and to compare this prodrug with the positive control drug doxorubicin. The study employed a UHLC-MS/MS method to determine the levels of albumin-bound of legubicin and two metabolites (free Leu-DOX and DOX) in plasma, tumor, and tissue samples. This method was validated for good selectivity, high sensitivity, excellent extraction recovery, and short run time. The results showed that legubicin was present in the circulation in vivo mainly in a protein-bound form with larger AUC values and lower clearance and distribution, and essentially released small amounts of doxorubicin. Compared to administration of equimolar doses of doxorubicin, legubicin showed increased exposure of the active drug in the tumor and decreased the level of the active drug in the heart and kidney. This study provides valuable information on the pharmacokinetics and tissue distribution of legubicin, implicating its potential as a novel and effective drug candidate for anti-cancer therapies.

Exosomes are tiny vesicles secreted by cells, with a diameter of 40-160 nm, which contain proteins, DNA, mRNA, long noncoding RNA, etc. Because of the low sensitivity and specificity of the conventional biomarkers for liver diseases, it is of utmost importance to discover novel, sensitive, specific, and non-invasive biomarkers. Exosomal long noncoding RNAs have been considered as potential diagnostic, prognostic, or predictive biomarkers in a wide range of liver pathologies. In this review, we discuss the recent progress on exosomal long noncoding RNAs that serve as potential diagnostic, prognostic, or predictive markers and molecular targets in patients with hepatocellular carcinoma, cholestatic liver injury, viral hepatitis, and alcohol-related liver diseases.

目的:探讨如何提升GLP试验机构应对新冠疫情所带来挑战的管理能力.方法:梳理中国、美国、OECD、英国的GLP监管政策,结合本机构2022年上海疫情实际经历,归纳GLP试验机构降低GLP遵从性的风险管控方法.结果与结论:GLP试验机构应着重预案能力的建设(如业务可持续性计划、灾难恢复计划),并从加强自身质量体系建设(如及时优化或新增标准操作规范)、强化技术人员培训等方面提升GLP试验机构管理水平,才能应对各类社会重大突发事件(如新冠疫情)和满足国内外监管机构的要求.

随着纳米材料的广泛应用,其生物安全性日益受到重视.其中,纳米氧化铁是应用较为广泛的医用纳米材料之一.研究显示,纳米氧化铁粒子会在细胞、亚细胞和分子生物学水平(如基因和蛋白水平)造成遗传损伤,其遗传毒性的潜在机制为进入细胞核后直接与DNA相互作用,引起DNA损伤.此外,还可通过氧化还原反应诱导活性氧产生,通过活性氧诱发DNA氧化损伤.本文简要介绍纳米氧化铁的分类及制备方法,并以常用的纳米零价铁和超顺磁纳米氧化铁为例,介绍纳米氧化铁的遗传毒性研究进展及潜在的致毒机制.

Female germline stem cells (FGSCs) have been found in mouse, rat, pig, sheep and human ovaries. However, there is no information on the isolation or long-term culture of FGSCs from non-human primates. Here, we identified the presence of FGSCs in the ovaries of juvenile (3-4-year-old) cynomolgus monkeys using DDX4 and Ki67 double immunofluorescence. Then, a long-term serum- and cell feeder-free culture system for these FGSCs was used to establish a cell line, and its biological characteristics were analyzed. We found that testosterone promoted self-renewal of the cells. This study confirmed for the first time the presence of FGSCs in the ovary of non-human primates. This culture system and cell line will be of great significance for research in medicine and reproductive biology.