Oncolytic bacteria represent a distinct group of microorganisms capable of selectively infecting and colonizing tumor tissues while sparing normal cells. They exert anti-tumor effects primarily by activating the host immune system and eliciting a robust anti-tumor immune response, which ultimately mediates tumor cell killing. According to their distinct mechanisms of action and target specificities, oncolytic bacteria are currently categorized into three main types: direct oncolytic bacteria, indirect oncolytic bacteria, and prophylactic oncolytic bacteria. However, the clinical translation of these therapies faces several challenges, including suboptimal immunogenicity and uneven spatiotemporal distribution in vivo, which hinder their broader application. This review systematically examines the primary bacterial species utilized in cancer therapy and addresses the key obstacles impeding their clinical translation. Our aim is to lay a theoretical foundation and offer a technical perspective for developing safer and more efficacious oncolytic bacterial strategies.

Therapeutic antibodies are valued for their high specificity and selectivity in immunotherapy. However, the potential toxicity they may elicit underscores the necessity of assessing their preclinical efficacy and safety using suitable animal models. In this context, we review the various categories and applications of humanized mice, which have been engrafted with human cells or tissues to mimic the human immune system. These models are extensively utilized in the nonclinical assessment and development of various antibody drugs, acting as a conduit to clinical research. However, several challenges remain, including the limited lifespan of humanized mice, inadequate engraftment of human cells, and the rudimentary nature of the immune environment in these models. The development of humanized immune system models in mice presents both opportunities and challenges, potentially leading to new insights into the evolution and application of antibody therapeutics.

Contezolid (MRX-I) is a novel ortho-fluorophenyl dihydropyridone developed by MicuRx Pharmaceuticals, Inc. It has been approved for the treatment of drug-resistant Gram-positive bacterial infections with relatively lower toxicity than other oxazolidinones such as linezolid. However, the toxicity profile has not yet been completely revealed. The aim of this study was to disclose the toxicity of contezolid in Sprague-Dawley (SD) rats and compare its toxicity profile with linezolid in a standard 4-week toxicity study. In this study, SD rats were orally administered with contezolid at doses of 20, 100, or 200/300 mg/kg/day for 28 consecutive days followed by a 28-day recovery period. Linezolid at doses of 100 or 200 mg/kg/day served as a comparator. Clinical observations, body weight, food consumption, hematology, clinical chemistry, urinalysis, and histopathological examinations were conducted. All females in the 200 mg/kg/day linezolid group were subjected to unscheduled death due to myelosuppression within the first 2 weeks. No abnormalities were noted in the 200 mg/kg/day contezolid group, and the dose level was escalated to 300 mg/kg/day from day 15. Myelosuppression or myelosuppression-associated effects were comparable between the 300-mg/kg/day contezolid group and the 100-mg/kg/day linezolid group. The ‘no observed adverse effect level’ (NOAEL) of contezolid was determined to be 100 mg/kg/day (with an average AUC0–24 h of 268.4 μg*h/mL). At the same dose levels, the toxicity of contezolid was significantly lower than that of linezolid. These findings demonstrate that contezolid exhibits a favorable safety profile compared with linezolid in this 4-week repeated-dose toxicity study in rats.

Introduction: Platelet glycoprotein (GP) Ibα is a key receptor for thrombosis. Under high shear conditions, GPIbα-VWF interactions are required for initiating platelet adhesion and vessel occlusion. GPIbα is also an important checkpoint for thrombo-inflammation in acute ischemic stroke. It has been considered as a desirable target against ischemic stroke for decades, but no anti-GPIbα drug has been successfully developed. Methods: CA1001 was humanized from our unique mAb crossing different species, and manufactured under GMP-like conditions with 99.9% purity. The efficacy of CA1001 was assessed using various in vitro platelet functional assays with blood samples and in vivo models, including state-of-the-art intravital microscopy thrombosis and transient middle cerebral artery occlusion (tMCAO) models. Pharmacokinetics (PK), pharmacodynamics (PD), and a 14-day regulatory toxicology study were conducted in rats and rhesus monkeys. Results: CA1001 specifically recognized platelet GPIbα from human, monkeys, rats, mice, rabbits and dogs. Using platelets from rhesus monkeys, healthy volunteers, and patients with peripheral artery disease, CA1001 dose-dependently inhibited ristocetin-induced platelet aggregation in vitro . Using laser injury and FeCl 3 injury intravital microscopy models, CA1001 inhibited thrombosis, prevented vessel occlusion, and importantly, promoted thrombus dissolution (thrombolysis) in vivo . In a 60-min tMCAO models, intravenous injection of CA1001 1 hour after tMCAO significantly reduced the cerebral infarct volume at 24 hours without increasing the risk of intracerebral hemorrhage. The PD studies showed that single bolus injection of CA1001 reached maximal anti-platelet effects within 5 minutes (0.25mg/kg in rats, 4mg/kg in monkeys) which was maintained following intravenous infusion. The extent and duration of the effect were dose-dependent. Plasma concentrations increased linearly with the dose received. In toxicology studies, CA1001 was well tolerated and safe without bleeding nor platelet count reduction. The No Obvious Adverse Event Level in rats and monkeys were 25mg/kg, and 100mg/kg respectively, which are 10 and 25 times the therapeutic targeted doses. Conclusion: The first-in-class humanized anti-GPIbα Fab CA1001 has potent anti-thrombotic effects, consistent PK/PD properties and favorable safety and tolerability profiles warranting further clinical development in healthy volunteers and patients with acute ischemic stroke.

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.

Cardiovascular diseases (CVD) are widely recognized as a leading cause of death worldwide; however, early diagnosis and disease progression monitoring play a crucial role in their clinical management. The common diagnostic and prognostic biomarkers have represented a revolutionary tool for studying CVD; however, their applications are limited to invasive irreversible heart diseases, such as drug-induced myocardial injury. In light of this information, a growing number of studies are currently investigating the diagnostic and prognostic potential of novel CVD biomarkers. Examples of this are long non-coding RNA (lncRNA) and other RNAs that are specifically expressed at the early stages of heart disease. These RNAs have been reported to be involved in the development of CVD via activating or inhibiting inflammatory mediators and angiogenesis-related factors, as well as endothelial cell proliferation, migration and phenotypic transformation. This review collectively summarizes the recent studies' results concerning exosomal lncRNA biogenesis, characterization, and function, as well as its role as a novel biomarker in a variety of CVD.

In recent years, significant breakthroughs have been made in the field of gene therapy. Adeno-associated virus (AAV) is one of the most promising gene therapy vectors and a powerful tool for delivering the gene of interest. Among the AAV vectors, AAV serotype 8 (AAV8) has attracted much attention for its efficient and stable gene transfection into specific tissues. Currently, recombinant AAV8 has been widely used in gene therapy research on a variety of diseases, including genetic diseases, cancers, autoimmune diseases, and viral diseases. This paper reviewed the applications and challenges of using AAV8 as a vector for gene therapy, with the aim of providing a valuable resource for those pursuing the application of viral vectors in gene therapy.

Exosomes, a type of extracellular vesicle, are commonly found in different body fluids and are rich in nucleic acids (circRNA, lncRNAs, miRNAs, mRNAs, tRNAs, etc.), proteins, and lipids. They are involved in intercellular communication. lncRNAs are responsible for the modulation of gene expression, thus affecting the pathological process of kidney injury. This review summarizes the latest knowledge on the roles of exosome lncRNAs and circulating lncRNAs in the pathogenesis, biomarker discovery, and treatment of chronic kidney disease, renal fibrosis, and acute kidney injury, providing an overview of novel regulatory approaches and lncRNA delivery systems.

Isoprenaline hydrochloride (IH) is a β-adrenergic receptor agonist commonly used in the treatment of hypotension, shock, asthma, and other diseases. However, IH-induced cardiotoxicity limits its application. A large number of studies have shown that long noncoding RNA (lncRNA) regulates the occurrence and development of cardiovascular diseases. This study aimed to investigate whether abnormal lncRNA expression is involved in IH-mediated cardiotoxicity. First, the Sprague-Dawley (SD) rat myocardial injury model was established. Circulating exosomes were extracted from the plasma of rats and identified. In total, 108 differentially expressed (DE) lncRNAs and 150 DE mRNAs were identified by sequencing. These results indicate that these lncRNAs and mRNAs are substantially involved in chemical cardiotoxicity. Further signaling pathway and functional studies indicated that lncRNAs and mRNAs regulate several biological processes, such as selective mRNA splicing through spliceosomes, participate in sphingolipid metabolic pathways, and play a certain role in the circulatory system. Finally, we obtained 3 upregulated lncRNAs through reverse transcription-quantitative PCR (RT-qPCR) verification and selected target lncRNA-mRNA pairs according to the regulatory relationship of lncRNA/mRNA, some of which were associated with myocardial injury. This study provides valuable insights into the role of lncRNAs as novel biomarkers of chemical-induced cardiotoxicity.

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.

细胞外囊泡(extracellular vesicles,EV)这一概念在2011年由国际细胞外囊泡协会(International Society for Extracellular Vesicles,ISEV)提出,其可作为治疗药物以及药物的递送载体广泛应用于疾病治疗的各个领域.基于EV的疗法可能成为一种除传统药物治疗和细胞治疗之外的疾病治疗新模式——EV无细胞疗法.作为载体,相比于病毒载体和合成非病毒载体,EV具有独特的优势,潜力巨大.但EV由于其特有的生物学性质,在临床转化中具有一定困难.且该领域相对较新,尚无专门针对基于EV疗法的相关政策和法规出台.本文通过ClinicalTrials.gov平台采集信息,总结基于EV疗法的研究进展,针对现有的监管体系提出建议,并对EV非临床研究的一般原则、药学研究、药效学和药代动力学研究及安全性评价等非临床评价策略进行探讨,为制定基于EV疗法的非临床评价研究方案提供参考.

通过与靶向信使RNA结合,小核酸药物能特异性地沉默疾病基因来治疗疾病.目前,已有12种小核酸药物在欧美国家上市,还有更多药物处于各个研发阶段.小核酸药物虽然来自人工合成,但因其自身特点不能单纯按照化学药物进行非临床评价.本文结合已上市药物的研究资料和相关指导原则,分别从成药性、药效学、药动学及安全性评价四方面对小核酸药物非临床特点和评价策略进行阐述.

Mulberry extract from Fructus Mori contains an anthocyanin pigment and has been widely used as a food additive in China and other Eastern Asian countries. Only few research has been done on toxicological profiling of mulberry extract for its safety evaluation; however, the data is inconclusive. In the current study, mulberry extract of 4200, 1400, or 466 mg/kg were orally administrated to Sprague Dawley rats for 90 consecutive days followed by a recovery period of 28 days. No abnormalities were detected in body weights, food intake, ophthalmological, hematological, coagulation, clinical chemistry, and organ weights parameters. Discoloration of urine (red, purple, and brown) and feces (black), along with bedding material (purple) were observed in the 4200 mg/kg group. Further, microscopic examination revealed brown granules in the renal tubular cells for rats in 4200 and 1400 mg/kg groups. Since these changes were associated with excretory effect of the extract, the No Observed Adverse Effect Level was determined to be 4200 mg/kg, which was equivalent to the 1058.5 mg/kg of anthocyanin.

目的 幼年SD大鼠连续182 d经口灌胃给予玉屏风颗粒浸膏(简称“玉屏风浸膏”),停药28 d,观察玉屏风浸膏对幼年SD大鼠发育毒性的影响.方法 160只SD大鼠随机分为4组,分别为阴性对照组(去离子水,0.00 g/kg)和玉屏风浸膏低、中及高剂量组,分别经口灌胃给予去离子水、3.95、7.90及15.80 g/kg玉屏风浸膏,等浓度(0.79 g/ml)不同体积;评价药物对动物生存情况、临床征状、体质量、摄食量、眼科、自发活动能力、学习和记忆功能、血液学指标、血清生化学指标和免疫功能、生长激素和胰岛素样生长因子Ⅰ、精子、骨密度和骨量指标以及组织病理学指标的影响.结果 与对照组相比,各给药组动物的生存情况、临床征状、体质量、摄食量、眼科、自发活动能力、学习和记忆功能、血液学指标、血清生化学指标和免疫功能、生长激素和胰岛素样生长因子Ⅰ、精子、骨密度和骨量指标以及组织病理学指标均无与给予玉屏风浸膏相关的明显毒性改变.结论 玉屏风浸膏在幼年SD大鼠中未观察到发育毒性的剂量水平为15.80 g/kg.

Cinobufotalin injection, a traditional Chinese medicine preparation, successfully used for several years, might induce cardiotoxicity. The aim of the study was to evaluate the cardiotoxicity of cinobufotalin injection and the cardiotoxicity-preventive effect of sodium phenytoin in vivo. According to the 4 × 4 Latin square design, four Beagle dogs were allocated into four dose levels of 0, 0.3, 1, and 3 g/kg in treatment phases I-IV (cinobufotalin injection) and 3 g/kg in treatment phase V (cardiotoxicity antidote). The following parameters and endpoints were assessed: clinical observations, body weight, indicators of myocardial injury, and electrocardiogram (ECG) parameters. The cinobufotalin injection-related changes were observed in clinical observations (rapid breathing pattern), indicators of myocardial injury (increased cardiac troponin I, creatine kinase isoenzymes, and aspartate aminotransferase), and ECG graphics (arrhythmia) at 3 g/kg concentration in treatment phases I-IV. The cardiotoxicity of cinobufotalin injection was attenuated by sodium phenytoin in treatment phase V. The results confirmed the cardiotoxicity of cinobufotalin injection, and they might bring information about the appropriate monitoring time points and cardiotoxicity parameters in clinical practices and shed light on the treatment of cardiovascular adverse reactions.

本试验研究了长链非编码RNA (lncRNA)在药物性肝损伤SD大鼠肝脏中表达谱的变化,采用lncRNA芯片对肝脏lncRNA表达谱进行分析,对原始数据进行归一化处理后,筛选出差异表达的lncRNA并通过基因功能GO富集、KEGG信号通路等生物信息学间接分析主要差异lncRNA的潜在功能.结果 表明,与正常SD大鼠肝脏相比,lncRNA表达谱发生了显著变化,2倍以上变化的lncRNAs共有902条(P＜0.05),其中上调的有566条,下调的lncRNA有336条,这说明差异性lncRNA可能参与了药物性肝损伤发生及发展过程.

间充质干细胞(mesenchymal stem cells,MSCs)因其具有特殊的低免疫原性和免疫调节能力,展现出良好的应用前景.但是近年来,MSCs产品的组织来源及体内移植途径的多样性,导致MSCs不良反应的报道越来越多.尤其是MSCs经血管输注过程中的栓塞问题越来越普遍.因此,本文就目前临床前和临床研究经血管输注MSCs后引起栓塞的因素进行总结,从而为临床前动物实验出现的栓塞风险如何正确有效预测临床上潜在风险提供思路.

安全药理学是新药非临床研究的重要组成部分,主要是研究药物剂量≥治疗剂量时,潜在的不期望出现的对生理功能的不良影响.近年来,随着分子生物学和生物技术等的进步,安全药理学的研究内容和研究范围有了较大的拓展.本文从国际国内安全药理学指导原则要求和修订,到中枢神经系统、心血管系统和呼吸系统安全药理及补充安全药理(胃肠道系统、泌尿系统、血液系统和肝胆系统)等方面,全面综述了新药非临床安全药理学研究进展.

Recent studies have shown that microRNA (miRNAs) can play important roles in the regulation of endothelial cell (EC) function. However, the expression profile of miRNAs and their effects on the apoptosis of ECs under microgravity conditions remains unclear. In this study, the apoptosis of human pulmonary microvascular endothelial cells (HPMECs) under simulated microgravity was identified by Annexin V and propidium iodide double staining and transmission electron microscopy. miRNA microarray assay was used to screen the differentially expressed miRNAs in HPMECs under simulated microgravity, and eight differentially expressed miRNAs were identified. Specifically, miR-503-5p, which was found to be most significantly upregulated in both microarray and quantitative reverse-transcription polymerase chain reaction assays, was selected for further functional investigation. Overexpression of miR-503-5p induced apoptosis of HPMECs under normal gravity and aggravated the negative effects of simulated microgravity on HPMECs. Furthermore, silencing of miR-503-5p expression effectively attenuated the negative effects of simulated microgravity on HPMECs. Further experiments showed that the mRNA and protein expression of anti-apoptotic factor B-cell lymphoma-2 (Bcl-2), which has been confirmed as a direct target of miR-503-5p, was inhibited by the upregulation of miR-503-5p and increased by the downregulation of miR-503-5p. Taken together, our findings demonstrate, for the first time, that miR-503-5p can induce apoptosis of HPMECs under simulated microgravity through, at least in part, inhibiting the expression of Bcl-2.