Quantitative Proteomics Reveals the Protective Effects of Huangqi Decoction Against Acute Cholestatic Liver Injury by Inhibiting the NF-κB/IL-6/STAT3 Signaling Pathway.

Intrahepatic cholestasis (IC) is a common syndrome that affects the liver, with treatment options being limited. Huangqi decoction (HQD), a classic herbal medicine, has shown protective effects against IC. In this study, isobaric tags for relative and absolute quantification-based quantitative proteomics was performed to investigate the potential mechanism of action of HQD on α-naphthylisothiocyanate (ANIT)-induced IC, resulting in 2796 quantified proteins across all samples, including 270 differentially expressed proteins under HQD treatment. Fuzzy c-means clustering analysis of these 270 proteins assigned the proinflammatory proteins, such as LCN2, SAA1, FGG, FGA, and FGB, to Cluster 1 (upregulated by ANIT, and downregulated by HQD). Functional bioinformatics and protein-protein interaction network analyses indicated that these proinflammatory proteins were involved in the STAT3 signaling pathway. Further real-time PCR and Western blot experiments confirmed that the expression of these proteins was consistent with the proteomic results. Moreover, HQD treatment decreased the phosphorylation of STAT3, induced by ANIT. Western blot experiments revealed that HQD treatment decreased phosphorylation of NF-κB and downregulated the expression of the inflammatory gene IL-6 and therefore inhibited the IL-6/STAT3 signaling pathway. In summary, the present study suggested that HQD may ameliorate acute cholestatic liver injury via inhibition of the NF-κB/IL-6/STAT3 signaling pathway.

Simultaneous quantification of multiple components in rat plasma by UPLC-MS/MS and pharmacokinetic study after oral administration of Huangqi decoction.

A rapid, sensitive and accurate UPLC-MS/MS method was developed for the simultaneous quantification of components of Huangqi decoction (HQD), such as calycosin-7-O-ß-d-glucoside, calycosin-glucuronide, liquiritin, formononetin-glucuronide, isoliquiritin, liquiritigenin, ononin, calycosin, isoliquiritigenin, formononetin, glycyrrhizic acid, astragaloside IV, cycloastragenol, and glycyrrhetinic acid, in rat plasma. After plasma samples were extracted by protein precipitation, chromatographic separation was performed with a C18 column, using a gradient of methanol and 0.05% acetic acid containing 4mm ammonium acetate as the mobile phase. Multiple reaction monitoring scanning was performed to quantify the analytes, and the electrospray ion source polarity was switched between positive and negative modes in a single run of 10 min. Method validation showed that specificity, linearity, accuracy, precision, extraction recovery, matrix effect and stability for 14 components met the requirements for their quantitation in biological samples. The established method was successfully applied to the pharmacokinetic study of multiple components in rats after intragastric administration of HQD. The results clarified the pharmacokinetic characteristics of multiple components found in HQD. This research provides useful information for understanding the relation between the chemical components of HQD and their therapeutic effects.

Increased systemic exposure to rhizoma coptidis alkaloids in lipopolysaccharide-pretreated rats attributable to enhanced intestinal absorption.

Rhizoma coptidis is a rhizome commonly used in traditional Chinese medicine. After oral administration of rhizoma coptidis extract, the plasma concentrations of its effective alkaloid constituents are so low that their systemic therapeutic actions cannot be explained. This study aimed to investigate the influence of lipopolysaccharide (LPS) on the pharmacokinetics of the rhizoma coptidis alkaloids. Pharmacokinetic experiments were performed with rats; both in vitro absorption and efflux experiments were carried out with everted rat gut sacs, whereas in vitro metabolism experiments were conducted with rat liver microsomes and intestinal S9 fractions. Mucosal changes were evaluated with light microscopy and transmission electron microscopy. The results showed that, in rat plasma, LPS pretreatment increased systemic alkaloid exposure. LPS pretreatment increased the in vitro absorption of the alkaloids and decreased their efflux. The efflux of vinblastine and rhodamine 123, P-glycoprotein substrates, also was decreased. The absorption of fluorescein isothiocyanate-labeled dextran (average molecular mass, 4 kDa), a gut paracellular permeability probe, was not influenced. Obvious damage was observed in the mucosa, but the tight junctions between epithelial cells remained intact. Intestinal, rather than hepatic, alkaloid metabolism was decreased. These findings indicated that LPS pretreatment increased systemic exposure to the alkaloids through enhancement of their absorption, which was related to decreased intestinal efflux and metabolism. The results add to the understanding of why rhizoma coptidis is active despite the low plasma concentrations of the rhizoma coptidis alkaloids measured in normal subjects and experimental animals.

Effect of a novel proteoglycan PTP1B inhibitor from Ganoderma lucidum on the amelioration of hyperglycaemia and dyslipidaemia in db/db mice.

Protein tyrosine phosphatase 1B (PTP1B) is implicated in the negative regulation of the insulin signalling pathway by dephosphorylating the insulin receptor (IR) and IR substrates. Ganoderma lucidum has traditionally been used for the treatment of diabetes in Chinese medicine; however, its anti-diabetic potency and mechanism in vivo is still unclear. Our previously published study reported a novel proteoglycan PTP1B inhibitor, named Fudan-Yueyang-Ganoderma lucidum (FYGL) from G. lucidum, with a half-maximal inhibitory concentration (IC50) value of 5·12 (sem 0·05) µg/ml, a protein:polyglycan ratio of 17:77 and 78 % glucose in polysaccharide, and dominant amino acid residues of aspartic acid, glycine, glutamic acid, alanine, serine and threonine in protein. FYGL is capable of decreasing plasma glucose in streptozotocin-induced diabetic mice with a high safety of median lethal dose (LD50) of 6 g/kg. In the present study, C57BL/6 db/db diabetic mice were trialed further using FYGL as well as metformin for comparison. Oral treatment with FYGL in db/db diabetic mice for 4 weeks significantly (P < 0·01 or 0·05) decreased the fasting plasma glucose level, serum insulin concentration and the homeostasis model assessment of insulin resistance. FYGL also controlled the biochemistry indices relative to type 2 diabetes-accompanied lipidaemic disorders. Pharmacology research suggests that FYGL decreases the plasma glucose level by the mechanism of inhibiting PTP1B expression and activity, consequently, regulating the tyrosine phosphorylation level of the IR ß-subunit and the level of hepatic glycogen, thus resulting in the improvement of insulin sensitivity. Therefore, FYGL is promising as an insulin sensitiser for the therapy of type 2 diabetes and accompanied dyslipidaemia.

Winemaking Characteristics of Red-Fleshed Dragon Fruit from Three Locations in Guizhou Province, China.

The aim of this study was to identify the locations and harvest months in Guizhou province, China, producing the most suitable red dragon fruit (Hylocereus polyrhizus) for winemaking. Fruit from Guanling, Luodian and Zhenfeng counties was harvested separately from successive fruit cycles in August, September and October, respectively. The key traits measured were fruit weight, pulp yield, soluble solids content, and titratable acid. Wine characteristics measured were alcohol content, total carbohydrates, titratable acidity, volatile acidity, and betacyanin content. The overall suitability of fruit from each location for winemaking was evaluated using a multi-factor, unweighted, scorecard. On that basis, fruit from Guanling county harvested in August was the most suitable. Fruit from Luodian, and Zhenfeng was most suitable when harvested in August and September, and September, respectively. These results provide a preliminary guide for the sourcing of red dragon fruit from Guizhou for wine production.

Yinchenzhufu decoction protects against alpha-naphthylisothiocyanate-induced acute cholestatic liver injury in mice by ameliorating disordered bile acid homeostasis and inhibiting inflammatory responses.

ETHNOPHARMACOLOGICAL RELEVANCE: Intrahepatic cholestasis is a common condition of many liver diseases with few therapies. Yinchenzhufu decoction (YCZFD) is a representative traditional Chinese herbal formula used for treating jaundice and liver disease. AIM OF THE STUDY: To investigate the hepatoprotective effect of YCZFD against cholestatic liver injury and reveal its potential mechanism. MATERIALS AND METHODS: Mice with alpha-naphthyl isothiocyanate (ANIT)-induced intrahepatic cholestasis were orally administered YCZFD at doses of 3, 6, and 12g crude drug/kg for 2 weeks followed by subsequent analyses. A serum metabolomics study was then performed to explore the different metabolites influenced by YCZFD using ultra-high-performance liquid chromatography coupled with linear ion trap-Orbitrap hybrid mass spectrometry (UPLC-LTQ-Orbitrap-MS/MS).The levels of individual bile acids in the serum, liver, and bile were determined by UPLC-MS/MS. The expression of metabolic enzymes, transporters, inflammatory factors, and cytokeratin-19 (CK-19) was determined by real-time PCR, western blotting, and immunohistochemistry. RESULTS: YCZFD administration decreased the serum biochemical indexes and ameliorated pathological damage, such as hepatic necrosis and inflammatory cell infiltration. Serum metabolomics revealed that the metabolites influenced by YCZFD were mainly associated with bile acid metabolism and inflammation. YCZFD administration effectively ameliorated the disordered bile acid homeostasis. The bile acid transporter, multidrug-resistance associated protein 2 (Mrp2), and the metabolic enzyme, cytochrome P450 2b10 (Cyp2b10), were upregulated in the YCZFD intervention group compared to those in the ANIT-induced group. YCZFD administration also significantly inhibited nuclear factor-κB (NF-κB) and its phosphorylation and decreased the expression of proinflammatory cytokines including tumor necrosis factor-α, interleukin-1ß, and intercellular adhesion molecule-1 in ANIT-induced cholestatic mice. Additionally, the level of CK-19 was lower in the YCZFD intervention group than in the ANIT-induced cholestatic mice. CONCLUSION: YCZFD administration ameliorated disordered bile acid homeostasis, inhibited NF-κB pathway-mediated inflammation, and protected the liver from bile duct injury. Therefore, YCZFD exerted a protective effect against cholestatic liver injury.

[The application of diminished criminal responsibility rating scale to mental retardation offenders].

OBJECTIVE: To explore the application of Diminished Criminal Responsibility Rating Scale (DCRRS) to mental retardation offenders. METHODS: The DCRRS was used to 121 cases of mental retardation offenders who were divided into three groups according to the degree of their diminished criminal responsibility. RESULTS: There were significant differences in rating score among the three groups (mild group 22.12+/-4.69, moderate group 25.50+/-5.48, major group 27.59+/-5.69), and 17 items had good correlation with the total score of the scale with the correlation coefficient from 0.289 to 0.665. Six factors were extracted by the factor analysis, and 69.392% variation could be explained. CONCLUSION: The DCRRS has rational items, its total score could show the difference among the three degree diminished criminal responsibility of mental retardation offenders.

[Advanced investigation of testamentary capacity of the mentally disordered].

Testamentary capacity is one of the civil competences, it means that a natural person enjoys the capacity or qualification to establish testament and deal with his property. Recently, the cases of testamentary capacity assessment of the mentally disordered are increasing. This article firstly introduces the concepts of the testament as well as the testamentary capacity, and then summarizes the assessment standard of the testamentary capacity, by using the Banks v. Goodfellow case as a basis to make the standard criteria including: the understanding of the nature of a will and codicil, the knowledge of the general extent of one's assets, the knowledge of the natural object of one's bounty, the understanding of the impact of the distribution of the assets of the estate, and the absence of a delusion specifically affecting the distribution of the estate. The impact factors of the testamentary capacity, including dementia, mood disorder, schizophrenia, alcohol, drug, and undue influence, etc., are summarized. Lastly, the related assessment tools such as the Mini-Mental State Examination, the Clock-Drawing Test, and the Testament Definition Scale are introduced briefly.

[Civil competence assessment of the mental disorders involved in compensation of personal injury].

OBJECTIVE: To seek and ascertain indicators that can be used in the civil competence assessment of the mental disorders involved in compensation of personal injury. METHODS: A retrospective study was made on the data related to the interviewee's mental status assessed by forensic experts during the period from 2003 to 2005 in Institute of Forensic Science, Ministry of Justice, P.R.China. The 6 indicators, including awareness of situation, factual understanding of issues, appreciation of likely consequences, rational manipulation of information, functioning in one's own environment, and communication of choice, were graded and statistically analyzed using SPSS 11.5 software. RESULTS: The 6 indicators correlated well with the assessment of forensic experts ,with the related coefficient between 0.632 and 0.876, and the inter-related coefficient among the 6 indicators between 0.575 and 0.911. CONCLUSION: The 6 indicators could be used for the civil competence assessment and may also be taken as the basis for further standardization and quantification of civil competence.

Protective effect of cultured bear bile powder against dimethylnitrosamine-induced hepatic fibrosis in rats.

Natural bear bile has been used for liver disease in East Asia for thousands of years. However, its use has restrictions. In the current study, the therapeutic effects and potential mechanisms of cultured bear bile powder (CBBP) against hepatic fibrosis were evaluated in a dimethylnitrosamine (DMN)-induced rat model. CBBP treatment significantly improved DMN-induced hepatic necrosis and inflammatory infiltration. Additionally, CBBP remarkably alleviated the increased hepatic collagen content and expression of alpha-smooth muscle actin. Serum metabolomics revealed that 14 serum metabolites, including docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were decreased in DMN-treated rats, which was reversed by CBBP. Pathway analyses revealed that the main metabolic pathways affected by CBBP were related to fatty acid biosynthesis and metabolism, and biosynthesis of unsaturated fatty acids. EPA and DHA are ligands of peroxisome proliferator activated receptors (PPARs). CBBP treatment significantly stimulated liver mRNA and protein expression of PPARα and PPARγ. CBBP also markedly increased liver expression of PPARα target genes, which are involved in fatty acid ß-oxidation, and down-regulated IL-6, a downstream inflammatory gene of PPARγ. In conclusion, CBBP has the potential to attenuate liver fibrosis and its mechanism involves the promotion of the liver expression of PPARα and PPARγ. Our results may help in the development of a novel substitute for bear bile and therapeutic strategies for fibrotic liver diseases.

Protective effect of herbal medicine Huangqi decoction against chronic cholestatic liver injury by inhibiting bile acid-stimulated inflammation in DDC-induced mice.

BACKGROUND: Huangqi decoction (HQD), a classic traditional herbal medicine, has been used for liver fibrosis, but its effect on intrahepatic chronic cholestatic liver injury remains unknown. PURPOSE: In the present study, we investigated the hepatoprotective effect of HQD and the underlying molecular mechanisms in 3, 5-diethoxycarbonyl-1, 4-dihydroxychollidine (DDC)-induced chronic cholestatic mice. METHODS: The DDC-induced cholestatic mice were administrated HQD for 4 or 8 weeks. Serum biochemistry and morphology were investigated. The serum and liver bile acid (BA) levels were detected by ultra performance liquid chromatography-tandem mass spectrometry. The liver expression of BA metabolizing enzymes and transporters, and inflammatory and fibrotic markers was measured by real-time polymerase chain reaction, western blotting, and immunohistochemistry. RESULTS: HQD treatment for 4 or 8 weeks ameliorated DDC-induced liver injury by improving impaired hepatic function and tissue damage. HQD treatment for 8 weeks further decreased the liver expression of cytokeratin 19, tumor growth factor (TGF)-ß, collagen I, and α-smooth muscle actin, and ameliorated ductular reaction and liver fibrosis. HQD markedly decreased the accumulation of serum and liver BA. The expression of BA-metabolizing enzymes, cytochrome P450 2b10 and UDP glucuronosyltransferase 1 A1, and multidrug resistance-associated protein 2, Mrp3, and Mrp4 involved in BA homeostasis was increased by 4 weeks of HQD treatment. The expression of BA uptake transporter Na+-taurocholate cotransporting polypeptide was decreased and that of Mrp4 was increased after 8 weeks of HQD treatment. Nuclear factor-E2-related factor-2 (Nrf2) was remarkably induced by HQD treatment. Additionally, HQD treatment for 8 weeks decreased the liver expression of inflammatory factors, interleukin (IL)-6, IL-1ß, tumor necrosis factor-α, monocyte chemoattractant protein-1, and intracellular adhesion molecule-1. HQD suppressed the nuclear factor (NF)-κB pathway. CONCLUSION: HQD protected mice against chronic cholestatic liver injury and biliary fibrosis, which may be associated with the induction of the Nrf2 pathway and inhibition of the NF-κB pathway, ameliorating BA-stimulated inflammation.

Rhodamine-based Hg2+-selective chemodosimeter in aqueous solution: fluorescent OFF-ON.

[reaction: see text] N-(Rhodamine-6G)lactam-N'-phenylthiourea-ethylenediamine (1) was developed as a fluorescent and colorimetric chemodosimeter in aqueous solution with a broad pH span (5 approximately 10) and high selectivity toward Hg2+ but no significant response toward other competitive cations, such as Fe2+, Co2+, Ni2+, Cu2+, Zn2+, Pb2+, Cd2+, Ca2+, Mg2+, K+, Na+, etc. The Hg2+-promoted ring opening of spirolactam of the rhodamine moiety induced cyclic guanylation of the thiourea moiety, which resulted in the dual chromo- and fluorogenic observation (OFF-ON).

Highly sensitive and selective chemosensor for Hg2+ based on the rhodamine fluorophore.

A novel tren-based tripodal chemosensor 1 bearing a rhodamine and two tosyl groups was synthesized and its sensing behavior toward metal ions was investigated by UV/vis and fluorescence spectroscopies. Addition of a Hg2+ ion to a CH3CN solution of 1 gave a visual color change as well as significantly enhanced fluorescence, while other ions including Pb2+, Zn2+, Cu2+, Ca2+, Ba2+, Cd2+, Co2+, Mg2+, Ag+, Cs+, Li+, and Na+ induced no or much smaller color/spectral changes, which constituted a Hg2+-selective fluorescent chemosensor (OFF-ON).

Fluorescence turn on of coumarin derivatives by metal cations: a new signaling mechanism based on C=N isomerization.

[reaction: see text] A new sensing mechanism based on C=N isomerization, which shows a very significant fluorescence enhancement to the metal cations in a simple and efficient way, is demonstrated. A coumarin derivative (L) containing a C=N group was designed as an example for illustration. The free ligand L is almost nonfluorescent due to the isomerization of C=N double bond in the excited state. However, the solution of ligand shows about a 200-fold increase of fluorescence quantum yield (about 30%) upon addition of Zn(ClO4)2.

An efficient chloride-selective fluorescent chemosensor based on 2,9-bis(4'-hydroxyphenyl)phenanthroline Cu(II) complex.

A new complex Cu(II)/L, composed of 2,9-bis(4'-hydroxyphenyl)phenanthroline (L) and Cu(II), was synthesized as an efficient chloride-detection fluorescent chemosensor with high selectivity and sensitivity over other halide anions, F(-), Br(-), I(-). The recognition mechanism was discussed primarily.

[Influencing factors of psychopath's civil litigation].

OBJECTIVE: To explore the influencing factors of schizophrenic patient's capability in civil litigation, and to establish the base of quantitative study about execution of civil litigation. METHODS: To study questionnaires completed from patients with and without civil litigation capabilities and to determine the influencing factors from medical and forensic aspects. RESULTS: One hundred patients were admitted to the study and were divided into two groups based on capability in civil litigation. There were significant differences in psychiatric and legal aspects between the groups. CONCLUSION: Capability of schizophrenic patients in perusing civil litigation had been impaired or even complete lost.

Huangqi Decoction Alleviates Alpha-Naphthylisothiocyanate Induced Intrahepatic Cholestasis by Reversing Disordered Bile Acid and Glutathione Homeostasis in Mice.

Intrahepatic cholestasis is a serious symptom of liver disorders with limited therapies. In this study, we investigated the efficacy of Huangqi decoction (HQD), a two-herb classic traditional Chinese medicine (TCM), in the treatment of alpha-naphthylisothiocyanate (ANIT)-induced intrahepatic cholestasis in mice. HQD treatment ameliorated impaired hepatic function and tissue damage. A metabolomics study revealed that the endogenous metabolites significantly affected by HQD were related to bile acid (BA) biosynthesis and glutathione metabolism pathways. HQD treatment decreased the intrahepatic accumulation of cytotoxic BAs, normalized serum BA levels, and increased biliary and urinary BA excretion. Additionally, HQD restored the hepatic glutathione content and suppressed reactive oxygen species (ROS) in cholestatic mice. Protein and gene analysis revealed that HQD increased the expression of the hepatic metabolizing enzymes cytochrome P450 (CYP) 2B10 and UDP glucuronosyltransferase family 1 member A1 (UGT1A1), as well as multidrug resistance-associated protein 2 (Mrp2), Mrp3, and Mrp4, which play crucial roles in BA homeostasis. Further, HQD increased the protein expression of glutamate-cysteine ligase, which is involved in the synthesis of glutathione. Importantly, HQD increased the nuclear expression of nuclear factor-E2-related factor-2 (Nrf2). In conclusion, HQD protects against intrahepatic cholestasis by reversing the disordered homeostasis of BAs and glutathione.

Chicken bile powder protects against α-naphthylisothiocyanate-induced cholestatic liver injury in mice.

This study explored the effects of chicken bile powder (CBP), a 2000-year-old Chinese medicine, on α-naphthyl isothiocyanate (ANIT)-induced intrahepatic cholestasis in mice. CBP treatment for 14 days significantly ameliorated ANIT-induced changes in serum alanine aminotransferase, aspartate aminotransferase, bile acids, bilirubin, γ-glutamyl transpeptidase, alkaline phosphatase, and liver tissue morphology. Serum metabolomics showed changes in 24 metabolites in ANIT-exposed mice; 16 of these metabolites were reversed by CBP treatment via two main pathways (bile acid biosynthesis and arachidonic acid metabolism). Additionally, CBP administration markedly increased fecal and biliary bile acid excretion, and reduced total and hydrophobic bile acid levels in the livers of cholestatic mice. Moreover, CBP increased liver expression of bile acid efflux transporters and metabolic enzymes. It also attenuated ANIT-induced increases in hepatic nuclear factor-κB-mediated inflammatory signaling, and increased liver expression of the nuclear farnesoid X receptor (FXR) in cholestatic mice. CBP also activated FXR in vitro in HEK293T cells expressing mouse Na+-taurocholate cotransporting polypeptide. It did not ameliorate the ANIT-induced liver injuries in FXR-knockout mice. These results suggested that CBP provided protection from cholestatic liver injury by restoring bile acid homeostasis and reducing inflammation in a FXR-dependent manner.

Novel fluorescent sensor for detection of Cu(II) in aqueous solution.

A novel fluorescent chemosensor based on aminonaphthol, which can selectively recognize copper(II) over other metal ions in aqueous solution within a broad pH span, was synthesized.

[The application of diminished criminal responsibility rating scale to schizophrenia offenders].

OBJECTIVE: To explore the feasibility of Diminished Criminal Responsibility Rating Scale(DCRRS) to schizophrenia offenders. METHODS: The DCRRS were used respectively to 325 cases of schizophrenia offender which were divided into three groups according to the degree of criminal responsibility. RESULTS: There were significant differences in rating score among three groups (mild group 21.16 +/- 4.17, moderate group 25.87 +/- 5.43, major group 29.35 +/- 4.60), and all items have good correlation with the total score of the scale. 6 factors were extracted by the factor analysis, and their cumulative squared loadings is 68.485%. CONCLUSION: The diminished criminal responsibility in schizophrenia offenders could be divided into three grades.