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Hepatic sinusoidal obstruction syndrome (HSOS) via exposure to pyrrolizidine alkaloids (PAs) is with high mortality and there is no effective treatment in clinics. Bear bile powder (BBP) is a famous traditional animal drug for curing a variety of hepatobiliary diseases such as cholestasis, inflammation, and fibrosis. Here, we aim to evaluate the protective effect of BBP against HSOS induced by senecionine, a highly hepatotoxic PA compound. Our results showed that BBP treatment protected mice from senecionine-induced HSOS dose-dependently, which was evident by improved liver histology including reduced infiltration of inflammatory cells and collagen positive cells, alleviated intrahepatic hemorrhage and hepatic sinusoidal endothelial cells, as well as decreased conventional serum liver function indicators. In addition, BBP treatment lowered matrix metalloproteinase 9 and pyrrole-protein adducts, two well-known markers positively associated with the severity of PA-induced HSOS. Further investigation showed that BBP treatment prevents the development of liver fibrosis by decreasing transforming growth factor beta and downstream fibrotic molecules. BBP treatment also alleviated senecionine-induced liver inflammation and lowered the pro-inflammatory cytokines, in which tauroursodeoxycholic acid played an important role. What's more, BBP treatment also decreased the accumulation of hydrophobic bile acids, such as cholic acid, taurocholic acid, glycocholic acid, as well. We concluded that BBP attenuates senecionine-induced HSOS in mice by repairing the bile acids homeostasis, preventing liver fibrosis, and alleviating liver inflammation. Our present study helps to pave the way to therapeutic approaches of the treatment of PA-induced liver injury in clinics.

Gynura japonica (also named Tusanqi in Chinese) is used as a folk herbal medicine for treating blood stasis or traumatic injury. However, hundreds of hepatic sinusoidal obstruction syndrome (HSOS) cases have been reported after consumption of preparations made from G. japonica because it contains large amounts of hepatotoxic pyrrolizidine alkaloids (PAs). To date, blood pyrrole-protein adducts (PPAs) are suggested as biomarkers for the diagnosis of PA-induced HSOS in clinics. However, the concentration of PPAs in the blood is greatly affected by several factors including the amount of PA exposure, herb intake period, and blood sampling time after the last exposure. In present study, the kinetic characters of PPAs in serum and liver as well as other potential target organs were studied systematically and comprehensively following multiple exposures of PAs in G. japonica extract (GJE). As results, PPAs content reached to a plateau both in serum and liver after the mice were treated with GJE for 2 weeks on daily basis. PPAs cleared significantly slower in liver (T1/2ke∼184.6 h, ∼7.7 days) than in serum (T1/2ke∼95.8 h, ∼4.0 days). Although more than 90 % PPAs were removed 2 weeks after the last dosing, PPAs still persisted in the liver until the end of the experiment, i.e. 8 weeks after the last dosing. The results would be of great help for understanding the importance of PPAs for PA-induced toxicity and its detoxification.

There is no gold standard for evaluating the severity of community-acquired pneumonia (CAP), and it is still based on a score. This study aimed to use the metabolomics method to find promised biomarkers in assessing disease severity and potential therapeutic targets for CAP. The result found that the metabolites in the plasma samples of CAP patients had significantly different between the acute phase and the remission phase, especially lysophosphatidylcholine (LPCs) in glycerophospholipids, whose levels are negatively linked to the severity of the disease. Subsequently, the two key metabolites of myristoyl lysophosphatidylcholine (LPC 14:0) and LPC 16:1 were screened. We analyzed the predictive performance of the two metabolites using Spearman-related analysis and ROC curves, and LPC14:0 showed more satisfactory diagnostic performance than LPC16:1. Then we explored the protective role and mechanism of LPC 14:0 in animal and cell models. The results showed that LPC 14:0 could inhibit the LPS-induced secretion of IL-1β, IL-6, and TNF-α, lower the ROS and MDA levels, and decreased the depletion of SOD and GSH, thereby reducing lung tissue and cell damage, such as down-regulating the protein level in BALF, lung W/D ratio, MPO activity, and apoptosis. We found that LPC 14:0 inhibited LPS-induced inflammatory response and oxidative stress, and the above protection was achieved by inhibiting LPS-induced activation of the NLRP3 inflammasome. LPC 14:0 may serve as a novel biomarker for predicting the severity of CAP. In addition, our exploration of the role of LPC 14:0 in animal and cellular models has reinforced its promise as a therapeutic target to improve the clinical efficacy for CAP.

Type 1 diabetes (T1D) causes cognitive decline and has been associated with brain metabolic disorders, but its potential molecular mechanisms remain unclear.The purpose of this study was to explore the molecular mechanisms underlying T1D-induced cognitive impairment using metabolomics and lipidomics.We developed an optimized integration approach of metabolomics and lipidomics for brain tissue based on UPLC-Q-TOF-MS and analyzed a comprehensive characterization of metabolite and lipid profiles in the hippocampus and frontal cortex of T1D male mice with cognitive decline (T1DCD) and age-matched control (CONT) mice.The results show that T1DCD mice had brain metabolic disorders in a region-specific manner relative to CONT mice, and the frontal cortex exhibited a higher lipid peroxidation than the hippocampus in T1DCD mice. Based on metabolic changes, we found that microglia was activated under diabetic condition and thereby promoted oxidative stress and neuroinflammation, leading to neuronal injury, and this event was more pronounced in the frontal cortex than the hippocampus.Our results suggest that brain region-specific shifts in oxidative stress and neuroinflammation may contribute to diabetic cognitive decline, and the frontal cortex could be the more vulnerable brain region than the hippocampus.

Application of organic amendments is an effective approach for improving soil organic carbon and soil fertility. To investigate the effects of different organic amendments on soil organic carbon and its labile fraction content, a batch of incubation experiments was conducted on the fluvo-aquic soil in Dongting Lake region, Hunan Province. There were six treatments, including soil amended with rice straw, soil amended with Chinese milk vetch, soil amended with bio-organic fertilizer, soil amended with pig manure, and soil amended with rice straw-derived biochar, with unamended soil as control. Each treatment had the same amount of carbon input. After 180 days of incubation, application of organic amendments increased soil labile organic carbon content. Application of bio-organic fertilizer, pig manure and rice straw-derived biochar significantly increased soil organic carbon content by 26.1%, 9.7% and 30.7%, respectively. There was no significant change in soil organic carbon content in rice straw and Chinese milk vetch treatments which were more favourable to the accumulation of soil dissolved organic carbon and microbial biomass carbon. Pig manure was more favourable to the accumulation of soil dissolved organic carbon. Bio-organic fertili-zer could benefit the accumulation of soil microbial biomass carbon and readily oxidizable organic carbon. Rice straw-derived biochar could promote the accumulation of soil microbial biomass carbon and light fraction organic carbon. Compared with rice straw, soil carbon pool management index was increased by 31.8%, 111.6%, 62.2% and 50.7% in Chinese milk vetch, bio-organic fertilizer, pig manure and rice straw-derived biochar treatments, respectively. The performance of bio-organic fertilizer, pig manure, and rice straw biochar was better than rice straw and Chinese milk vetch from the perspective of soil carbon sequestration and soil carbon pool management index.有机物料还田是提升农田土壤有机碳、培肥土壤的重要措施。为探讨不同有机物料的还田效果,采用室外培养方法,研究了在等碳输入条件下,施用水稻秸秆、紫云英、生物有机肥、猪粪和水稻秸秆生物炭对洞庭湖双季稻区潮土有机碳和活性有机碳组分含量的影响。结果表明: 经过180 d的培养试验,与不施用有机物料相比,施用有机物料提高了土壤活性有机碳含量。生物有机肥、猪粪和水稻秸秆生物炭处理分别使土壤有机碳含量显著提升了26.1%、9.7%和30.7%,水稻秸秆和紫云英对土壤有机碳含量的提升效应在试验期间并不显著。水稻秸秆和紫云英还田更有利于土壤可溶性有机碳和微生物生物量碳的积累,猪粪更有利于土壤可溶性有机碳的积累,生物有机肥更有利于土壤微生物生物量碳和易氧化有机碳的积累,水稻秸秆生物炭则更有利于土壤微生物生物量碳和轻组有机碳的积累。与水稻秸秆还田相比,紫云英、生物有机肥、猪粪和水稻秸秆生物炭还田使土壤碳库管理指数分别提高了31.8%、111.6%、62.2%和50.7%。从土壤固碳和土壤碳库管理指数来看,生物有机肥、猪粪和水稻秸秆生物炭的还田效果优于水稻秸秆和紫云英还田。.

Pyrrolizidine alkaloids (PAs) are natural toxins found in about 3%-5% of flowering plants. Dehydropyrrolizidine alkaloids contain a double bond in 1, 2-position of the necine bases, including retronecine type PAs (RET-PAs) and their N-oxides (RET N-oxide-PAs), and otonecine type PAs (OTO-PAs), and are known for their significant hepatotoxicity. Most dehydropyrrolizidine alkaloids are metabolically activated by cytochrome P450 (CYP450) enzymes to generate active pyrroles, which further bind to proteins to form pyrrole-protein adducts (PPAs). Methods for predicting PA-induced liver injury are generally performed on in vitro models with extremely low activities of CYP450 enzymes, which is different from the situation in vivo. In this regard, primary cultured mouse hepatocytes, which showed comparable CYP450 activity with the in vivo models, were applied to illustrate the structure-toxicity relationship of 13 dehydropyrrolizidine alkaloids, namely, eight RET-PAs, three RET N-oxide-PAs, and two OTO-PAs. PA-induced cytotoxicity and PA-generated PPAs were analyzed in primary mouse hepatocytes treated with different PAs. Results showed that PA-induced toxicity was correlated with the amount of PA-generated PPAs. RET-PAs and OTO-PAs were generally more toxic than RET N-oxide-PAs and generated higher amount of PPAs. PPAs were utilized to evaluate the efficiency of metabolic activation and predict the toxic potencies of dehydropyrrolizidine alkaloids. The proposed model could be a new approach for toxicity evaluation and risk control of exposure to PAs.

Pyrrolizidine alkaloids (PAs) are naturally occurring plant toxins associated with severe liver damage if excessive ingestion. Herein, a novel analytical strategy on utilizing direct analysis in real-time mass spectrometry (DART-MS) was developed, and applied in analysis of six representative PAs. The calibration curves in the range of 10–1000 ng·mL−1 were established, and relative standard deviations (RSDs) were less than 10%. The limits of detection (LODs) and limits of quantitation (LOQs) were 0.55–0.85 ng·mL−1 and 1.83–2.82 ng·mL−1, respectively. The feasibility of method was indicated by analysing real samples including Gynura japonica, drug tablets, granules, and fresh cow’s milk. Moreover, the results of DART-MS were in good agreement with those observed by high performance liquid chromatography mass spectrometry (HPLC-MS), but consumed less time without chromatographic separation. This research provides a facile fashion for safety assessment of herbal and food products containing PAs and presents promising applications in food, pharmaceutical and clinical analysis.

Rat hippocampal neurons were isolated and divided into Normal, oxygen glucose deprivation/reoxygenation (OGD/R), OGD/R + DEX, OGD/R + NC mimic, OGD/R + miR-155 mimic and OGD/R + DEX + miR-155 mimic groups. In OGD/R group, LDH, ROS and MDA levels and apoptosis rate was increased, with up-regulations of miR-155, Cyt c and Bax/Bcl-2 ratio, but decreases of SOD, GSH-Px and MMP levels, as well as down-regulations of p-ERK1/2/ERK1/2. As compared to the OGD/R group, parameters above in the OGD/R + DEX group were ameliorated evidently, while OGD/R + miR-155 mimic group manifested the opposite changes. Besides, miR-155 mimic could abolish the protective effect of DEX on the hippocampal neurons under OGD/R. DEX, via down-regulating the expression of miR-155, could activate the ERK1/2 pathway, thereby mitigating the apoptosis and oxidative stress injury and increasing the MMP, thereby protecting hippocampal cells from OGD/R injury.

MicroRNAs (miRNAs) - as negative regulators of target genes - are associated with various human diseases, but their precise role(s) in diabetic retinopathy (DR) remains to be elucidated. The aim of this study was to elucidate the involvement of miRNAs in early DR using in silico analysis to explore their gene expression patterns.We used the streptozotocin (STZ)-induced diabetic rat to investigate the roles of miRNAs in early DR. Retinal miRNA expression profiles from diabetic versus healthy control rats were examined by miRNA array analysis. Based on several bioinformatic systems, specifically, gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, we identified signatures of the potential pathological processes, gene functions, and signaling pathways that are influenced by dysregulated miRNAs. We used quantitative real-time polymerase chain reaction (qRT-PCR) to validate six (i.e. those with significant changes in expression levels) of the 17 miRNAs that were detected in the miRNA array. We also describe the significant role of the miRNA-gene network, which is based on the interactions between miRNAs and target genes.GO analysis of the 17 miRNAs detected in the miRNA array analysis revealed the most prevalent miRNAs to be those related to biological processes, olfactory bulb development and axonogenesis. These miRNAs also exert significant influence on additional pathways, including the mitogen-activated protein and calcium signaling pathways. Six of the seventeen miRNAs were chosen for qRT-PCR validation. With the exception of a slight difference in miRNA-350, our results are in close agreement with the differential expressions detected by array analysis.This study, which describes miRNA expression during the early developmental phases of DR, revealed extensive miRNA interactions. Based on both their target genes and signaling pathways, we suggest that miRNAs perform critical regulatory functions during the early stages of DR evolution.

Conclusion: This study indicates that the lesion of hair cells in the apical turn of the cochlea can cause the change in the summating potential (SP)/Compound potential (CAP) ratio.Objectives: Electrocochleography is a valuable clinic test for diagnosis of cochlear pathologies and the ratio of SP to CAP has been used to identify Meniere’s disease. However, it remains controversial whether the increase of the SP/CAP ratio represents exclusively the endolymphatic hydrops.Method: This study measured the SP and CAP in mice that displayed outer hair cell (OHC) degeneration in the apical section of the cochlea as their age increased.Results: As compared with the mice aged 8–10 months, the 24-month old mice displayed a significant increase in the amplitude of SP at 12–16 kHz. This result suggests that the degeneration of OHCs in the apical turn leads to the increase of the + SP at the middle frequencies. In contrast, the aging mice did not have a significant change in the CAP amplitude at super-threshold...

The neural mechanisms that give rise to hyperacusis, a reduction in loudness tolerance, are largely unknown. Some reports suggest that hyperacusis is linked to childhood hearing loss. However, the evidence for this is largely circumstantial. In order to rigorously test this hypothesis, we studied loudness changes in rats caused by intense noise exposure (12 kHz narrow band noise, 115 dB SPL, 4 h) at postnatal 16 days. Rats without noise exposure were used as controls. The exposed noise group (n = 7) showed a mean 40-50 dB hearing loss compared to the control group (n = 8) at high frequencies (>= 8 kHz) and less hearing loss at lower frequencies. Loudness was evaluated using sound reaction time and loudness response functions in an operant conditioning-based behavioral task using narrow-band noise (40-110 dB SPL, centered at 2, 4 and 12 kHz). Interestingly, the sound reaction time of the noise group was significantly shorter than the control group at supra-threshold levels. The average reaction time was less than 100 ms in the noise group at 100 dB SPL, which was three times shorter than the control group. Our results indicate that early noise-induced hearing loss leads to a significant increase of loudness, a behavior indicative of hyperacusis. Our results are consistent with clinical reports suggesting that hearing loss at an early age is a significant risk factor for hyperacusis.

Vascular endothelial growth factor (VEGF) is one of the major regulatory molecules in diabetic retinopathy (DR). In our previous study, we demonstrated that succinate levels were elevated in the retinas of diabetic rats and that the knockdown of the succinate receptor, G-protein-coupled receptor 91 (GPR91), inhibited the release of VEGF and attenuated retinal vascular disorder in the early stages of DR. In the present study, we examined the signaling pathways involved in the GPR91-dependent release of VEGF in the retinal ganglion cell line, RGC-5. The cells were infected with a lentiviral small hairpin RNA (shRNA) expression vector targeting GPR91 (LV.shGPR91). Immunofluorescence staining revealed that GPR91 was predominantly localized in the cell bodies of the RGC-5 cells. RT-qPCR, western blot analysis and ELISA indicated that succinate exposure upregulated VEGF expression, activated the extracellular signal-regulated protein kinase (ERK)1/2, c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) signaling pathways and led to the release of cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2). The knockdown of GPR91 inhibited ERK1/2 and JNK activity, but did not inhibit the activation of the p38 MAPK pathway. The increase in COX-2 expression and the release of PGE2 were inhibited by transduction with LV.shGPR91 and ERK1/2, JNK and COX-2 inhibitors. The expression and release of VEGF showed similar results. Cell Counting Kit-8 (CCK-8) assays revealed that the shRNA-mediated knockdown of GPR91 decreased the proliferation of RF/6A cells cultured in succinate-conditioned medium. Our data suggest that GPR91 modulates the succinate-induced release of VEGF through the MAPK/COX-2/PGE2 signaling pathway.

To investigate the effects of urokinase-type plasminogen activator (uPA) on the reproductive function of the male rats with ornidazole-induced infertility.Fifty 10-12 weeks old adult male Sprague-Dawley rats were randomly divided into five groups: low-dosage uPA (330 IU/[kg x d]), mid-dosage uPA (1000 IU/[kg x d]), high-dosage uPA (3000 IU/[kg x d]), ornidazole (400 mg/[kg x d]) and control (0.5% Carboxymethylcellulose solution). The ornidazole group was treated by gastric gavage, and the rats in the uPA groups given both ornidazole by gastric gavage and uPA by intraperitoneal injection at the same time. All the rats were treated for 20 days consecutively, followed by copulation experiment. The rats were sacrificed and the reproductive system explored.The percentage of motile sperm and the number of embryos in the high-dosage uPA group increased significantly (P0.01) compared with the ornidazole group.uPA can antagonize ornidazole-induced infertility in male rats. The effect might be attributed to the improvement of sperm motile function by uPA.

To observe the effects of water decoction of the root of Crataegus cuneata on infertility induced by multi-glucoside of Tripterygium wilfordii (GTW) in rats.Male adult rats were randomly divided into five groups, which were treated via gastric gavage of distilled water (1 mL x kg(-1)) , solution of GTW (10 mg x kg(-1)) and three doses of water decoction of root of C. cuneata (1.8, 5.4, 18 g x kg(-1)) + GTW (10 mg x kg(-1)), respectively. 8 weeks later, GTW was stopped and the decoction and water continued for another 4 weeks. And then, all the male rats were copulated with adult female rats. The rates of pregnancy, average numbers of embryos and luteum of female rats, relative weights of reproductive organs, sperm counts, sperm motility and viability were compared among all the groups. The histology and ultrastructure of testis and epididymis were observed, while the concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH) and testostorone (T) in serum and T in testicular homogenate were detected by radioimmunoassay.Compared with those in GTW model group, the embryo numbers, the relative weight of testis and epididymis and sperm counts and motility in C. cuneata groups were increased obviously (P0.05). After treatment, the morphological damages of seminiferous tubules and sperms were recovered, while concentrations of T in testicular homogenate were also significantly increased (P0.01).C. cuneata could relieve the reproductive lesions induced by GTW, and hence improve the uberty of the male infertile model rats.

The orphan nuclear receptor steroidogenic factor-1 (SF-1) regulates the expression of several genes involved in the reproductive function and development of the adrenal, the gonads, and the pituitary gonadotropes. It also confers the gonadotrope-specific expression of the glycoprotein hormone a subunit gene by the binding to a gonadotrope-specific element (GSE). In this study, we have shown that SF-1 transactivates the salmon gonadotropin II beta subunit (sGTHII beta) gene expression. SF-1 alone offered a slight but significant enhancement on sGTHII beta promoter activity (7.2 +/- 0.6 fold). However, it stimulated sGTHII beta gene expression dramatically (127 +/- 37 fold) when combined with the estrogen receptor (ER). This synergistic interaction was specific for sGTHII beta promoter as well as for both SF-1 and ER and was estradiol-dose dependent. 5'-Deletion studies of the sGTHII beta promoter identified two putative SF-1 binding sites (GSE) and one previously identified proximal estrogen-responsive element (pERE) at -274 bp involved in this activation. The two GSE sequences located at -354 bp (sGSE(3) and -162 bp (sGSE(2) upstream of the transcription site, although imperfect as compared with the consensus GSE, bound specifically to the in vitro-translated mouse SF-1 protein. 5'-Deletion studies, competition experiments, and site-directed mutagenesis showed that binding to pERE and GSE(2) were necessary for the SF-1/ER synergistic effect. These studies suggest that the synergistic interaction of SF-1 and ER, possibly through cooperative binding or protein-protein interaction, is essential in conferring a cell type-specific expression of the GTHII beta subunit gene.