A large number of studies have shown that long non-coding RNAs (IncRNAs) display abnormallity in organisms exposed to toxic chemicals,carcinogens and heavy metals.In this paper,the relationships between IncRNAs and microRNAs (miRNAs),the expressions of IncRNAs in organisms exposed to different exogenous toxic chemicals and the related toxicological mechanism are reviewed in order to provide reference for biological monitoring with IncRNAs in environmental toxicology.

Drug-induced liver injury(DILI) is one of the major causes of termination of drug development.The establishment of a high-throughput test system to predict potential clinical hepatotoxicity is a valuable approach in the pharmaceutical industry.The high-content imaging-based in vitro assays allow simultaneous detection of cellular multiple parameters in the system.The real-time monitoring of multiple signaling pathways can shed light on many mechanisms of cell injury,with high sensitivity and specificity.Many types of liver cell models have been applied to high-content screening(HCS) so far.This paper introduceds the HCS technology and reviews the data of hepatotoxicity obtained from HCS technology in recent years.At the same time,we discuss the application of this technology in exploring the mechanism of hepatotoxicity and the potential of HCS technology in studying DILI and mechanisms.

Arsenic trioxide (As2O3) has been considered a poison,which is also known as an old drug and has recently been re-introduced as a new medicine.As2O3 shows potent effect on many types of cancers,especially on a specific types of leukemia-acute promyelocytic leukemia (APL).This poison drug As2O3 is effective against all stages of APL and has been approved by the Food and Drug Administration (FDA) of the United States for the treatment of APL.However,the clinical use of As2O3 has been limited by its toxicities,especially cardiotoxicity.This review focuses on the therapeutic effect on APL and the side effect during treatment.

OBJECTIVE To study the absorption,distribution and excretion of 2-fluorine-6-trifluoromethylpyridine (JJBD) in rats.METHODS [14C] Radioactivity isotope tracing method was used.Male SD rats were ig given a single dose of JJBD 10 and 100 mg·kg-1 (radioactivity:3.7 GBq·kg-1).Concentrations of rat plasma,tissue,feces,urine and bile were determined with a liquid scintillation counting (LSC) analyzer.Toxicokinetics (TK) parameters were fitted using WinNonlin.RESULTS TK parameters of JJBD 10 and 100 mg · kg-1 in male SD rats were as follows:area under the curve (AUC（0-t）) was 22 548±1579 and (203 395±27 586) h·iμg Eq.·L-1,half time (t1/2) was 15.8±1.0 and (14.1±0.9) h,peak time (Tmax) was 4.0±3.0 and (6.0±5.0) h,peak concentration (Cmax) was 1450±355 and (7776±1703) μg Eq.·L-1.JJBD was mainly distributed in fat,livers,kidneys,stomachs and intestinal walls.The concentration of JJBD in most of the tissues reached peak values after 4 h.However,JJBD couldn't be detected in the muscle,thymus gland,brain,gonad or spleen.Excretion rate of JJBD was 43.1％ in urine,29.7％ in feces and 9.97％ in cleaning solution within 0-168 h.JJBD could be excreted through bile at a rate of 28.1％ within 0-72 h.CONCLUSION JJBD can be absorbed immediately and excreted slowly in SD rat.There is no accumulation risk.The distribution of JJBD in vivo is very extensive,but cannot go through the blood-brain barrier.JJBD is mostly excreted through feces and urine.

OBJECTIVE To explore the toxic effects of zinc oxide nanoparticls (ZnO-NPs) on cardiac development of zebrafish embryos and rat myocardial cell lines (H9c2),as well as potential molecular mechanisms.METHODS ZnO-NPs were characterized.Zebrafish embryos were exposed to different doses of ZnO-NPs (0,0.5,2.5,5.0 and 10.0 mg· L-1) for 24 to 96 h at 4 h post fertilization (4 hpf).The embryo mortality was observed.The expressions of notochord homeobox (noto),T-box 6 (tbx16),T,brachyury homolog a(ta),and tbx6 which were related to cardiac mesoderm were investigated using real-time PCR at 17 hpf.The heart rate and number of cardiomyocytes of embryos [[Tg (cmlc2:nucdsRed)] exposed to 0,2.5 and 5 mg· L-1 ZnO-NPs were detected at 72 hpf.Rat myocardial cell lines (H9c2) were treated with ZnO-NPs (0.1,0.5,1.0,5.0 and 10.0 mg· L-1) for 24 h.Cell viability was measured with Alamar Blue method.Mitochondrial ultrastructure was observed by transmission electron microscopy.Cellular ATP was detected using chemiluminescence,and oxygen consumption rate (OCR) was examined with Seahorse instrument.RESULTS The particle size of ZnO-NPs was (331 ±3)nm.The ZnO-NPs LC50 of zebrafish embryos at 48 hpf was 21.81 mg· L-1.The mRNA expressions of noto,ta and tbx6 were reduced after ZnO-NPs 2.5 mg· L-1 treatment at 17 hpf.The heart rate of 72 hpf zebrafish was 153 min-1 in the ZnO-NPs 5 mg· L-1 group,12.6％ lower than that in the cell vehicle group (P＜0.01),and the number of cardiomyocytes decreased by 15.5％ (P＜0.01) compared with the cell vehicle group.Reduced cell viability and mitochondrial vacuolation were observed in H9c2 after ZnO-NPs 0.5 mg· L-1 exposure.Compared with the cell vehicle group,the cell ATP decreased by 25.7％ (P＜0.05),and OCR decreased by 27.2％ (P＜0.01).CONCLUSION Low-dose ZnO-NPs exposure has effect on the cardiac development of zebrafish,mainly due to reduced heart rate and decreased number of cardiomyocytes.These changes may be related to the decreased expressions of cardiac development-related genes and the impairment of mitochondrial structure and function.

OBJECTIVE To evaluate the embryo toxicity of Shuanghuanglian (SHL) by the combination of a human placental barrier model and embryonic stem (ES) cell test model.METHODS A human placental barrier model was set up by placenta slice culture and Ussing chamber.SHL 0.2,0.4,0.8,1.6,3.2,6.4 and 12.8 g· L-1 was added into the maternal side of the human placental model,respectively.All the media was collected respectively from the matemal side and fetal side 60 min later and taken as the SHL containing medium.ES cells (D3 line) and embryonic fibroblast cells (BALB/c 3T3) were cultured with the SHL containing medium respectively from the maternal side and the fetal side for 10 d.Cell viability was detected by MTT assay,and 50％ survival inhibitory ratio of ES and 3T3 cells by SHL was calculated.ES cells were incubated with the SHL containing medium from the matemal side or fetal side when they differentiated to cardiac myoblasts using hanging drop-suspension-attachment method.Messenger RNA of myosin heavy chain genes (β-MHC) was detected by Q-PCR for differentiation ratio,and 50％ differentiation inhibitory ratio of ES cells by SHL was calculated.A statistics formula was used for prediction of SHL embryotoxicity potential.RESULTS The IC50 of SHL in the matemal side of the human placental model for 3T3 cell survival,ES cell survival and ES differentiation was 1.97,0.84 and 0.48 g· L-1,respectively.According to the criteria for embryo toxicity evaluation,SHL had weak embryo toxicity.However,the IC50 of SHL in the fetal side of the human placental model for 3T3 cell survival,ES cell survival and ES differentiation was 3.19,2.57 and 0.95 g· L-1,respectively.According to the criteria for embryo toxicity evaluation,the supernatant containing SHL that went through the placental barrier had no embryo toxicity.CONCLUSION SHL is safe in the test concentration range during pregnancy.It is more scientific to evaluate embryo toxicity of drugs by ES cell test with the samples obtained through the placental barrier during pregnancy.

OBJECTIVE To study the toxicity of zinc oxide nanoparticles (ZnO-NPs) on murine macrophage Ana-1 cells and the mechanism.METHODS Ana-1 cells were incubated with ZnO-NP (2.5-160 mg· L-1).Cell viability was investigated by MTT assay.The integrity of cell membrane was investigated by acridine orange-ethidium bromide (AO-EB) staining.The intracellular uptake of ZnO-NP and the percentage of sub-G1 of Ana-1 cells were detected by flow cytometry.Zinc ions were determined by fluorescent probe.The change of cell viability was studied after chelating zinc ions with ethylene diamine tetraacetic acid (EDTA).RESULTS ZnO-NP 2.5,5,10 and 20 mg· L-1 decreased cell viability of Ana-1 cells (r=0.905,P＜0.05) in a concentration-dependent manner.The cell viability was decreased to 27.9％ after exposure to ZnO-NP 20 mg· L-1.Intracellular uptake of ZnO-NP was increased after Ana-1 cell incubated with ZnO-NP at concentrations ranging from 40 to 160 mg· L-1 (P＜0.05).There were obvious free zinc ions in the cells.EDTA 2.5 mmol· L-1 significantly increased the cell viability decreased by ZnO-NP 20 mg· L-1 (P＜0.05).Chelating free zinc ions significantly mitigated ZnO-NP induced cell toxicity (P＜0.05).CONCLUSION Cytotoxicity and apoptosis of Ana-1 cells induced by ZnO-NP might be related to intracellular uptake of ZnO-NP and release of zinc ions.

OBJECTIVE To investigate the effect of prenatal nicotine exposure on cardiac ejection function and myocardial fibrosis of the offspring of rats.METHODS Pregnant rats were sc given nicotine 6.0 mg· kg-1,once daily for 17 d.The body mass and heart mass of the offspring were detected at the 21th day of gestation,and 15 and 90 d after birth.Heart rate of 90 d offspring was recorded by ECG,and cardiac functions were detected by Doppler ultrasonography,including cardiac output (CO),stroke volume (SV),ejection fraction (EF),left ventricular long axis shortening fraction (FS),interventricular septum diastolic diameter (IVSd) and left ventricular posterior wall diastolic diameter (LVPWd).The myocardial ultrastructure was detected under an electron microscope.Masson staining was used to detect the myocardial collagen fiber deposition.The level of collagen protein type Ⅰ in heart tissue was detected by radioimmunoassay.RESULTS Compared with control group,prenatal nicotine exposure resulted in a decrease of heart mass and body mass in groups of 21 d fetal rats and 15 d offspring(P＜0.05,P＜0.01),but had no effect on the 90 d offspring.Compared with the normal control group,the heart rate of 90 d offspring increased [366+10 vs (418+10) min-1] (P＜0.05),CO,FS and EF decreased (P＜0.01),and IVSd and LVPWd increased (P＜0.05,P＜0.01).Electron microscopy revealed that in the heart of nicotine 90 d offspring,myocardial fiber arrangement was loosened and confused,while extracellular matrix increased.Masson staining showed collagen deposited in the myocardium.The level of collagen type Ⅰ in heart tissue increased [0.59±0.09 vs (0.40±0.05) tμg·g-1 tissue] (P＜0.01).CONCLUSION Prenatal nicotine exposure induces the increased level of cardiac collagen type Ⅰ,myocardial fibrosis and decrease of cardiac ejection function in adult offspring,which may lead to increased susceptibility to cardiovascular diseases.

OBJECTIVE To investigate the mechanism of nicotinamide mononucleotide (NMN) on inflammation and fibrosis between endogenous nicotinamide phosphoribosyltransferase (NAMPT) and bone morphogenetic protein 7 (BMP-7) in diabetic glomerular cells.METHODS ① In vivo,spontaneous diabetic C57/BL6 mice and wild C57/BL6 mice were divided into two groups.When blood glucose was above (34.2±1.9) mmol· L-1,renal histology of diabetic mice became obvious.The protein expressions of Nampt and nuclear transcription factors-kappa B p65 (NF-κB p65),silent mating type information regulation 2 homolog 1 (SIRT1) and BMP7 were analyzed by lengths of immunofluorescence.② In vitro,rats' glomerular cells HBZY-1 were incubated with glucose 200 mmol· L-1 for different lengths of time (0,24,48 and 72 h) and at different concentrations of NMN (0,50,100 and 200 iμmol· L-1).The protein levels of Nampt and BMP7 were detected by Western blotting and the protein expressions of NF-κB p65 and α-SMA were measured by immunofluorescence assay.The protein levels of Nampt,BMP7 and NF-κB p65 were detected by Western blotting after HBZY-1 cells were treated with NMN 100 μmol· L-1 and FK866 10 μmol· L-1 for 24 h.RESULTS ① In vivo,the glomeruli of diabetic C57/BL6 mice showed obvious atrophy.Fluorescence intensity of Nampt was increased (P＜0.05),but that of BMP7 and SIRT1 in renal glomeruli cells was decreased compared with the wild type (P＜0.01).② In vitro,HBZY-1 cells were cultured in glucose 200 mmol· L-1 for 48 and 72 h.The protein expression of NAMPT was increased,but that of BMP7 was decreased (P＜0.05,P＜0.01).Expressions of NF-κB p65 and α-SMA were increased (P＜0.01) by immunofluorescence.The expression of BMP7 was increased after treatment with glucose 200 mmol· L-1,followed by NMN 50,100 and 200 μmol · L-1 for 24 h (P＜0.01).The expressions of NAMPT and NF-κB p65 were decreased (P＜0.01).The expressions of Nampt and NF-κB p65 in glucose 5.6 mmol· L1 +FK866 and glucose 5.6 mmol· L-1+ NMN groups were increased (P＜0.01),but the expression of BMP7 did not change.CONCLUSION Upregulation of endogenous Nampt obviously intervenes in BMP7 expression in the process of glomerular inflammatory fibrosis in severe diabetes.NMN can affect the protein expression of BMP7 via a special Nampt signaling pathway.

OBJECTIVE To investigate the effect of Turkish galls extract (TGE) on the expression of IgA in serum,urine and renal tissue of IgA nephropathy (IgAN) model rats.METHODS Fifty healthy male Sprague Dawley rats were randomly divided into normal control group,IgAN model group,and TGE 75,150 and 300 mg· kg-1 groups,10 rats per group.The model of IgAN rats was established with bovine serum albumin (BSA)+lipopolysaccharide (LPS)+carbon tetrachlorid (CCl4)for 12 weeks.From the 13th week,TGE was ig administrated once a day for 4 weeks.At the end of the 12th and 16th weeks,24 h urine protein was measured by BCA method.At the end of the 16th week,serum and urinary IgA levels were measured by enzyme linked immunosorbent assay(ELISA),serum creatinine(SCR) and blood urea nitrogen (BUN) were detected by an automatic biochemical analyzer,and the renal pathological changes were evaluated with an Oxford classification scoring system.The deposition of IgA immune complex in the kidney was observed by immunofluorescence assay.RESULTS At the end of 12th week,24 h urine protein increased in all IgAN groups (P＜0.05),compared with normal control group.At the end of 16th week,24 h urine protein,IgA content in serum and urine,SCr and BUN content in serum,score in Oxford classification of renal tissue and deposition of IgA immune complex in the kidney in IgAN model group were all higher than in normal control group (P＜0.05).Compared with IgAN model group,24 h urine protein,IgA content in serum and urine and SCr content in serum were decreased in all TGE groups (P＜0.05),and BUN content in serum and deposition of IgA immune complex in the kidney decreased in TGE 150 and 300 mg·kg-1 groups (P＜0.05).The score in Oxford classification of renal tissue was decreased in TGE 300 mg· kg-1 group only.CONCLUSION TGE has curative effect on IgAN model rats by reducing serum and urinary IgA and decreasing IgA immune complex deposition.