Inactivation of Heparin by Cationically Modified Chitosan

This study was performed to evaluate the ability of N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride (HTCC), the cationically modified chitosan, to form biologically inactive complexes with unfractionated heparin and thereby blocking its anticoagulant activity. Experiments were carried out in rats in vivo and in vitro using the activated partial thromboplastin time (APTT) and prothrombin time (PT) tests for evaluation of heparin anticoagulant activity. For the first time we have found that HTCC effectively neutralizes anticoagulant action of heparin in rat blood in vitro as well as in rats in vivo. The effect of HTCC on suppression of heparin activity is dose-dependent and its efficacy can be comparable to that of protamine-the only agent used in clinic for heparin neutralization. HTCC administered i.v. alone had no direct effect on any of the coagulation tests used. The potential adverse effects of HTCC were further explored using rat experimental model of acute toxicity. When administered i.p. at high doses (250 and 500 mg/kg body weight), HTCC induced some significant dose-dependent structural abnormalities in the liver. However, when HTCC was administered at low doses, comparable to those used for neutralization of anticoagulant effect of heparin, no histopathological abnormalities in liver were observed.     

Trema micrantha toxicity in horses in Brazil

After ingesting green leaves of T. micrantha, 2 horses showed apathy, locomotor deficit, blindness, recumbency, paddling, coma and death. The main gross findings were scattered haemorrhages, enhanced lobular pattern of the liver, and cerebral oedema. Histological changes included disseminated haemorrhages, massive hepatocellular necrosis, neuronal degeneration, Alzheimer type II astrocytes and cerebral perivascular oedema. Clinicopathological findings which were comparable with those observed in Trema micrantha poisoned ruminants, associated with epidemiological evidence suggested the diagnosis.Trema micrantha poisoning should be evaluated as a possible cause in the diagnosis of equine hepatopathy and occasional secondary encephalopathy.     

基于肝毒性探讨何首乌临床合理应用

目的 促进临床合理使用何首乌,降低肝毒性不良反应发生率。方法 通过查阅文献及相关实验数据分析,探索何首乌肝毒性发生的潜在因素,并针对因素提出临床合理使用建议和用药监护。结果 何首乌肝毒性的发生与炮制方法、炮制程度、化学成分、特异体质等因素有关,临床使用时应注重药品质量,辨证用药,对易感人群加强监护。结论 合理使用何首乌,可降低肝毒性不良反应发生率。     

Comparative Proteomic Analysis Shows an Elevation of Mdhl Associated with Hepatotoxicity Induced by Copper Nanoparticle in Rats

Copper nanoparticle is a new material widely used in biological medicine, animal husbandry and industrial areas, but its potential toxicity to human health and environment remains unclear. In order to study the hepatotoxic mechanisms of nanoparticles copper, two-dimensional gel electrophoresis （2-DE） and matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI/TOF MS） of proteomics technology were used to isolate and identify the differentially expressed proteins from liver, which associated with hepatotoxicity induced by copper nanoparticle in rats. In this study, we have screened 15 kinds of proteins related with hepatotoxicity, of which spot8212 was identified as Malate dehydrogenase （Mdhl）. The mRNA expression trend of Mdhl was consistent with the result of 2-DE by RT-PCR validation. Bioinformatics analysis showed that Mdhl was stable and no signal peptides, subcellular location was in endoplasmic reticulum; it contained many functional sites such as malate dehydrogenase activity signal sites 155LTRLDHNRAKSQI167; a helixes and random coils were the two main elements. Homologous analysis demonstrated high homologous of Mdhl in rats with mouse and human, and the phylogenetic tree of Mdhl was constructed. The result indicated that copper nanoparticle could regulate up the Mdhl protein expression so as to compensate the energy deficit. Energy metabolic disturbance may be a pathway for copper nanoparticle particles to exert the hepatotoxic effects in rats.     

Prevalence of elevated alanine transaminase activity in dogs treated with CCNU (Lomustine)*

The purpose of this study was to evaluate prevalence of serum alanine transaminase (ALT) elevation in dogs receiving lomustine (CCNU) and to analyse the pattern of occurrence and potential risk factors. Serum ALT activity in 109 dogs during single‐agent CCNU chemotherapy was retrospectively analysed. The median initial dose, dose‐intensity and cumulative dose of CCNU were 64 mg m^?2, 21 mg m^?2 week^?1 and 171 mg m^?2, respectively. The overall prevalence of major ALT elevation [> 5‐fold upper reference limit (URL)] was 29% (32/109) and developed most commonly after one to three doses of CCNU. These ALT elevations occurred without preceding mild ALT elevation in 53% (17/32) of the cases. Three dogs (2.8%) developed clinical hepatopathy. For severe ALT elevation (>10‐fold URL), age ≤5‐year‐old was associated with higher risk. The findings of this study showed that elevation of ALT is common during CCNU chemotherapy in dogs and severe elevation can develop on a sudden onset.     

姜黄素对草鱼急性肝损伤的保护作用研究

以CCl_4诱导的草鱼(Ctenopharyngodon idellus)急性肝损伤模型为基础,考察姜黄素对肝脏组织的保护作用。对草鱼腹腔注射不同剂量CCl_4-橄榄油后,分别于不同时间点测定血清中谷丙转氨酶(GPT)、谷草转氨酶(GOT)活性,确定CCl_4-橄榄油最佳注射剂量和作用时间;以建立的草鱼急性肝损伤模型为基础,采用不同浓度姜黄素(5、25、75、150、300 mg/kg)进行灌胃给药,观察草鱼血清和肝脏组织中相关生化指标变化情况。结果表明,用15%的CCl_4-橄榄油腹腔注射(剂量为0.1 m L/10 g鱼体重)24 h后,使草鱼血清中GPT、GOT水平升高(P0.05);同时,150 mg/kg姜黄素能显著(P0.05)抑制血清中GOT和GPT水平的升高。姜黄素对CCl_4诱导的草鱼肝组织损伤具有保护作用。     

The potential of organoids in toxicologic pathology: role of toxicologic pathologists in in vitro chemical hepatotoxicity assessment

The development of in vitro toxicity assessment methods using cultured cells has gained popularity for promoting animal welfare in animal experiments. Herein, we briefly discuss the current status of hepatoxicity assessment using human- and rat-derived hepatocytes; we focus on the liver organoid method, which has been extensively studied in recent years, and discuss how toxicologic pathologists can use their knowledge and experience to contribute to the development of in vitro chemical hepatotoxicity assessment methods for drugs, pesticides, and chemicals. We also propose how toxicological pathologists should assess toxicity regarding the putative distribution of undifferentiated and differentiated cells in the organoid when liver organoids are observed in hematoxylin and eosin–stained specimens. This was done while considering the usefulness and limitations of in vitro studies for toxicologic pathology assessment.     

Deep learning-based image-analysis algorithm for classification and quantification of multiple histopathological lesions in rat liver

Artificial intelligence (AI)-based image analysis is increasingly being used for preclinical safety-assessment studies in the pharmaceutical industry. In this paper, we present an AI-based solution for preclinical toxicology studies. We trained a set of algorithms to learn and quantify multiple typical histopathological findings in whole slide images (WSIs) of the livers of young Sprague Dawley rats by using a U-Net-based deep learning network. The trained algorithms were validated using 255 liver WSIs to detect, classify, and quantify seven types of histopathological findings (including vacuolation, bile duct hyperplasia, and single-cell necrosis) in the liver. The algorithms showed consistently good performance in detecting abnormal areas. Approximately 75% of all specimens could be classified as true positive or true negative. In general, findings with clear boundaries with the surrounding normal structures, such as vacuolation and single-cell necrosis, were accurately detected with high statistical scores. The results of quantitative analyses and classification of the diagnosis based on the threshold values between “no findings” and “abnormal findings” correlated well with diagnoses made by professional pathologists. However, the scores for findings ambiguous boundaries, such as hepatocellular hypertrophy, were poor. These results suggest that deep learning-based algorithms can detect, classify, and quantify multiple findings simultaneously on rat liver WSIs. Thus, it can be a useful supportive tool for a histopathological evaluation, especially for primary screening in rat toxicity studies.     

miR‐122‐5p as a plasma biomarker of liver injury in fish exposed to microcystin‐LR

Recent studies have shown the presence of large amounts of microRNAs (miRNAs; miRs) from damaged cells in the peripheral blood. In this study, we investigated the levels of miRNAs circulating in the blood plasma of whitefish (Coregonus lavaretus) after exposure to microcystin‐LR. We used real‐time PCR to examine the relative expression of plasma levels of 4 miRNAs (miR‐122‐5p and let‐7c‐5p, the liver‐enriched microRNAs, miR‐148a‐3p which promotes the hapatospecific phenotype in mammals, and miR‐92a‐3p, a cell proliferation and angiogenesis promoter, potentially hepatocarcinogenic) during the first 48 h after exposure to MC‐LR. We observed a rapid increase of miR‐122‐5p levels 8 h after exposure (P < 0.05), which continued to the end of the experiment. Our results demonstrated that the plasma miR‐122‐5p was indicative of MC‐LR‐induced liver injury, exhibiting areas under the curve close to 1 in ROC analysis (AUC = 0.976, P < 0.001). Although plasma levels of miR‐148a‐3p and miR‐92a‐3p were significantly elevated by the end of the experiment, their discriminative power was lower than reported for the miR‐122‐5p. Based on these results and reports on miRNA‐based diagnosis of liver injuries in mammals, plasma miR‐122‐5p could be considered as a robust, new generation diagnostic biomarker in fish, helpful for the non‐invasive diagnosis of liver damage.     

芽孢杆菌发酵五味子提取液对肉鸡肝损伤的影响

为探索中药与益生菌联用治疗肉鸡肝损伤的新方法,从而改善畜禽＂亚健康＂,本研究以CCl4注射肉鸡,建立肝损伤模型,并利用芽孢杆菌发酵五味子提取液,将发酵提取液混饮肝损伤肉鸡,检测外周血谷草转氨酶（Aspar-tate transaminase,AST）、谷丙转氨酶（Alanine aminotransferase,ALT）、过氧化物岐化酶（Superoxide dismutase,SOD）、丙二醛（Malondialdehyde,MDA）、谷胱甘肽还原酶（Glutathione reductase,GR）含量,并进行组织病理学观察。结果显示,芽孢杆菌在五味子提取液与培养基1∶1（V/V）混合液中生长良好,五味子有效成分在发酵前后含量无明显变化,发酵提取液按0.5%、1%混饮7d后,与肝损伤阳性对照组相比,2个用药组AST（P〈0.05）、ALT（P〈0.05）、SOD（P〈0.05）、GR（P〈0.01）含量显著升高,MDA（P〈0.05）含量显著降低,肝脏病变较轻。结果表明,芽孢杆菌发酵五味子提取液对提高机体抗氧化能力具有一定的作用,能够治疗肉鸡肝损伤。