Dose-related liver injury of Geniposide associated with the alteration in bile acid synthesis and transportation.

Fructus Gardenia (FG), containing the major active constituent Geniposide, is widely used in China for medicinal purposes. Currently, clinical reports of FG toxicity have not been published, however, animal studies have shown FG or Geniposide can cause hepatotoxicity in rats. We investigated Geniposide-induced hepatic injury in male Sprague-Dawley rats after 3-day intragastric administration of 100 mg/kg or 300 mg/kg Geniposide. Changes in hepatic histomorphology, serum liver enzyme, serum and hepatic bile acid profiles, and hepatic bile acid synthesis and transportation gene expression were measured. The 300 mg/kg Geniposide caused liver injury evidenced by pathological changes and increases in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and γ-glutamytransferase (γ-GT). While liver, but not sera, total bile acids (TBAs) were increased 75% by this dose, dominated by increases in taurine-conjugated bile acids (t-CBAs). The 300 mg/kg Geniposide also down-regulated expression of Farnesoid X receptor (FXR), small heterodimer partner (SHP) and bile salt export pump (BSEP). In conclusion, 300 mg/kg Geniposide can induce liver injury with associated changes in bile acid regulating genes, leading to an accumulation of taurine conjugates in the rat liver. Taurocholic acid (TCA), taurochenodeoxycholic acid (TCDCA) as well as tauro-α-muricholic acid (T-α-MCA) are potential markers for Geniposide-induced hepatic damage

Simulation of magnetic field distribution of excitation coil for EM flow meter and its validation using magnetic camera

Distribution of magnetic flux density in electromagnetic (EM) flow meter is significantly important for obtaining accurate flow rate. Analysis of Helmholtz coils used to generate homogeneous magnetic flux density in the cross section of flow pipe of electromagnetic (EM) flow meter is presented in this paper. In this work a simulation model with the EM module of finite element analysis software, COMSOL, is built and experimental testing of the distribution of magnetic flux density is applied to verify its uniformity. A 2D magnetic camera system with 64 Hall sensors is prototyped. The magnetic field data are picked up by the National Instruments PCI 6255 data acquisition card.The results from the simulation and measurements are presented and compared in the paper. The study helps to design the electrode sensor arrays of EM flow meter. In addition to this, the distribution data of magnetic flux density are useful for the improvement of accurate flow rate measurement of EM flow meter

KSR-Based Medium Improves the Generation of High-Quality Mouse iPS Cells

<div><p>Induced pluripotent stem (iPS) cells from somatic cells have great potential for regenerative medicine. The efficiency in generation of iPS cells has been significantly improved in recent years. However, the generation of high-quality iPS cells remains of high interest. Consistently, we demonstrate that knockout serum replacement (KSR)-based medium accelerates iPS cell induction and improves the quality of iPS cells, as confirmed by generation of chimeras and all iPS cell-derived offspring with germline transmission competency. Both alkaline phosphatase (AP) activity assay and expression of Nanog have been used to evaluate the efficiency of iPS cell induction and formation of ES/iPS cell colonies; however, appropriate expression of Nanog frequently indicates the quality of ES/iPS cells. Interestingly, whereas foetal bovine serum (FBS)-based media increase iPS cell colony formation, as revealed by AP activity, KSR-based media increase the frequency of iPS cell colony formation with Nanog expression. Furthermore, inhibition of MAPK/ERK by a specific inhibitor, PD0325901, in KSR- but not in FBS-based media significantly increases Nanog-GFP⁺ iPS cells. In contrast, addition of bFGF in KSR-based media decreases proportion of Nanog-GFP⁺ iPS cells. Remarkably, PD can rescue Nanog-GFP⁺ deficiency caused by bFGF. These data suggest that MAPK/ERK pathway influences high quality mouse iPS cells and that KSR- and PD-based media could enrich homogeneous authentic pluripotent stem cells.</p></div

Effects of FBS and small molecules on reprogramming in KSR medium.

<p>(A–C) IPS cells derived from MEFs. Representative FACS images (upper) and the percentage of Nanog-GFP⁺ cells by FACS analysis (lower) in different FBS/KSR ratio media on day 12 post-infection (A). AP⁺ colony number (B) and the percentage of Nanog-GFP⁺ cells (C) in different induction media on day 12. Induction media were KSR, FBS, or KSR based media with different small molecules (PD, CH, SB, and NAB, respectively). (D–F) IPS cells derived from adult fibroblasts. Schematic of the induction process for 12 days in different media series. There were 10 groups with different series of culture media. Each box represented 2 day period using indicated medium (MEF medium: green, KSR medium: blue, FBS medium: yellow). On day 3, the same number of infected fibroblasts were replated on new dishes using the appointed media. Cells were monitored and counted on day 12 (right column was Nanog-GFP⁺ colony number/total colony number). The mean values were derived from three parallel experiments (D). AP⁺ colony number (E) and the percentage of Nanog-GFP⁺ cells (F) in different induction media on day 12 post-infection. ** represented significant differences between KSR and other conditions, ∧ represented significant differences between FBS and other conditions (P<0.01).</p

KSR improves the quality of iPS cells derived from MEFs.

<p>(A) Morphology and Nanog-GFP fluorescence in primary iPS cells on day 12 in the FBS or KSP induction media. (B–E) Comparison of primary iPS cells between FBS group and KSR group on day 12 using different assays, including the following: AP staining (B), Nanog-GFP⁺ FACS (C), statistical analysis of the AP⁺ colony number and of the percentage of Nanog-GFP⁺ cells (D), and endogene expression by qPCR (E). (F–G) qPCR analysis of expression of pluripotent endogenes and exogenes in iPS cell lines induced in KSR medium or in FBS medium at passage 5. Normal mouse ES cells (NF2) were used as a positive control, whereas MEFs used as a negative control.</p

PD rescues repressive effect of bFGF in iPS cell induction.

<p>(A–B) Comparison of primary iPS cells derived from adult fibroblasts among different induction media on day 12 using different assays, including AP staining (A upper), the percentage of Nanog-GFP⁺ cells by FACS (A lower), and the number of Nanog-GFP⁺ colonies by fluorescence microscope (B). The induction media were FBS based and FBS with PD medium (FBS+PD); KSR medium and KSR medium with small molecules, including bFGF (4 ng/ml), bFGF (4 ng/ml) + PD, bFGF (0.4 ng/ml), and bFGF (0.4 ng/ml) + PD, respectively. (C) Functional model of MAPK/ERK pathway in the induction process. FGF may be one of the barriers in FBS that blocks formation of high-quality iPS cells. PD0325901, which selectively inhibits the MAPK/ERK pathway, benefits activation of Nanog and pluripotent endogenes during reprogramming.</p

Ameliorative effect of cheqianzi decoction on hyperuricemia and kidney injury and underlying mechanism in rats

Cheqianzi Decoction (CQD) is a Traditional Chinese Medicine (TCM) formula comprising four herbs and is recorded in the Ancient Materia Medica “Shengji Zonglu”. Individually, these four herbs have been shown to reduce uric acid (UA) levels, to treat hyperuricemia (HUA), and alleviate kidney damage. However, the therapeutic efficacy of the CQD and related mechanism are not yet clear.In this study, high performance liquid chromatography (HPLC) analysis confirmed that the contents of the chemical components of the four herbal medicines were in accordance with the provisions of the Chinese Pharmacopoeia. A total of 99 potential targets were identified in the network pharmacology analysis of CQD, indicating its involvement in the regulation of inflammatory and apoptotic signaling pathways, and potential value for treating HUA and alleviating kidney injury. In vivo pharmacodynamic studies showed that compared with the Model group, significantly decreased levels of serum uric acid (SUA), serum creatinine (SCr), blood urea nitrogen (BUN) (all P < 0.05), and inflammatory factors (P < 0.01) were detected in the CQD group. Quantitative real-time PCR and Western blot analyses showed that compared with the Model group, adenosine triphosphate (ATP)-binding cassette efflux transporter G2 (ABCG2) expression in the CQD group was significantly upregulated (P < 0.01) at both the mRNA and protein levels, while mRNA expression of Caspase3 and NOD-like receptor family member 3 (NLRP3) (P < 0.05) and protein expression of NLRP3 (P < 0.01) were significantly downregulated.In conclusion, CQD promotes UA excretion by activating ABCG2, and induces inflammasome NLRP3-mediated reduction in inflammatory and apoptotic factors to achieve renal protection. Thus, our findings indicate the therapeutic potential of CQD in HUA with kidney injury

Effects of Rhein on Bile Acid Homeostasis in Rats

Rhein, the active ingredient of rhubarb, a medicinal and edible plant, is widely used in clinical practice. However, the effects of repeated intake of rhein on liver function and bile acid metabolism are rarely reported. In this work, we investigated the alterations of 14 bile acids and hepatic transporters after rats were administered with rhein for 5 weeks. There was no obvious injury to the liver and kidney, and there were no significant changes in biochemical indicators. However, 1,000 mg/kg rhein increased the liver total bile acid (TBA) levels, especially taurine-conjugated bile acids (t-CBAs), inhibited the expression of farnesoid X receptor (FXR), small heterodimer partner (SHP), and bile salt export pump (BSEP) mRNA, and upregulated the expression of (cholesterol 7α-hydroxylase) CYP7A1 mRNA. Rhein close to the clinical dose (10 mg/kg and 30 mg/kg) reduced the amounts of TBAs, especially unconjugated bile acids (UCBAs), and elevated the expression of FXR and multidrug resistance-associated protein 3 (Mrp3) mRNA. These results denote that rhein is relatively safe to use at a reasonable dose and timing. 30 mg/kg rhein may promote bile acid transport and reduce bile acid accumulation by upregulating the expression of FXR mRNA and Mrp3 mRNA, potentially resulting in the decrease in serum UBCAs