Bowel preparation protocol for hospitalized patients ages 50 years or older: A randomized controlled trial.

BACKGROUND: The incidence and mortality rate of colorectal cancer progressively increase with age and become particularly prominent after the age of 50 years. Therefore, the population that is ≥ 50 years in age requires long-term and regular colonoscopies. Uncomfortable bowel preparation is the main reason preventing patients from undergoing regular colonoscopies. The standard bowel preparation regimen of 4-L polyethylene glycol (PEG) is effective but poorly tolerated. AIM: To investigate an effective and comfortable bowel preparation regimen for hospitalized patients ≥ 50 years in age. METHODS: Patients were randomly assigned to group 1 (2-L PEG + 30-mL lactulose + a low-residue diet) or group 2 (4-L PEG). Adequate bowel preparation was defined as a Boston bowel preparation scale (BBPS) score of ≥ 6, with a score of ≥ 2 for each segment. Non-inferiority was prespecified with a margin of 10%. Additionally, the degree of comfort was assessed based on the comfort questionnaire. RESULTS: The proportion of patients with a BBPS score of ≥ 6 in group 1 was not significantly different from that in group 2, as demonstrated by intention-to-treat (91.2% vs 91.0%, P = 0.953) and per-protocol (91.8% vs 91.0%, P = 0.802) analyses. Furthermore, in patients ≥ 75 years in age, the proportion of BBPS scores of ≥ 6 in group 1 was not significantly different from that in group 2 (90.9% vs 97.0%, P = 0.716). Group 1 had higher comfort scores (8.85 ± 1.162 vs 7.59 ± 1.735, P < 0.001), longer sleep duration (6.86 ± 1.204 h vs 5.80 ± 1.730 h, P < 0.001), and fewer awakenings (1.42 ± 1.183 vs 2.04 ± 1.835, P = 0.026) than group 2. CONCLUSION: For hospitalized patients ≥ 50 years in age, the bowel preparation regimen comprising 2-L PEG + 30-mL lactulose + a low-residue diet produced a cleanse that was as effective as the 4-L PEG regimen and even provided better comfort.

Physicians' knowledge and sepsis guide implementation in tertiary care hospitals in China.

BACKGROUND: This study was designed to investigate the extent to which physicians involved in sepsis management understand and adopt sepsis guidelines in clinical practice. The overarching aim of this study was to generate ideas for developing more effective training methods to help physicians apply the guidelines in patient management. METHODS: Physicians working in a tertiary care hospital, primarily in the emergency and critical care departments, were recruited into the survey. They were asked to fill questionnaires which were designed to collect sepsis score, diagnostic indicators, fluid resuscitation, antibiotics choice, access to knowledge and training, as well as implementation of sepsis guidelines in clinical diagnosis and treatment. RESULTS: Overall, the response rate was 625/661 (94.5%). The investigate shows the basic information of all physicians who participated in the answer sheet, including their work department, professional title and whether their hospital was a teaching hospital. Significant differences were identified among the physicians in terms of method of acquiring sepsis guidelines, the impact of study guidelines on clinical diagnosis and treatment, efficiency of training methods, cognition of fluid resuscitation in patients with sepsis, the cognition of sepsis rehydration principles, selection of antibiotics for patients with sepsis, the basis for antibiotic selection, among other variables. CONCLUSION: Although majority of physicians involved in tertiary care hospital understand the contents of sepsis-3 guidelines, the clinical implementation of the guidelines in the diagnosis and treatment of patients with sepsis is highly heterogeneous. Thus, there is need to develop standardized training for physicians involved in sepsis diagnosis and treatment.

Lysozyme Improves the Inhibitory Effects of Panax notoginseng Saponins on Phenotype Transformation of Vascular Smooth Muscle Cells by Binding to Ginsenoside Re.

Panax notoginseng saponins (PNS) have been used to treat cardiovascular diseases for hundreds of years in China. Lysozyme can bind to exogenous compounds and promote their activity. Nevertheless, knowledge of whether there is a synergistic role between lysozyme and PNS is far from sufficient. In this study, we show that the mixture of PNS and lysozyme synergistically inhibited platelet derived growth factor BB (PDGF-BB)-induced vascular smooth muscle cell (VSMC) viability, and in the five main components of PNS, GS-Re, but not GS-Rb1, NG-R1, GS-Rg1, or GS-Rd, reduced VSMC viability by combined application with lysozyme. Next, the supramolecular complexes formed by GS-Re and lysozyme were detected by mass spectrometry, and the binding ability increased with the concentration ratio of GS-Re to lysozyme from 4:1 to 12:1. In the supramolecular complexes, the relative contents of α-helix of lysozyme were increased, which was beneficial for stabilizing the structure of lysozyme. The 12:1 mixture of GS-Re and lysozyme (12.8 µmol/L GS-Re+1.067 µmol/L lysozyme) repressed PDGF-BB-induced VSMC viability, proliferation, and migration, which were associated with the upregulated differentiated markers and downregulated dedifferentiated markers. Finally, in CaCl2-induced rodent abdominal aortic aneurysm (AAA) models, we found that the 12:1 mixture of GS-Re and lysozyme slowed down AAA progression and reversed phenotype transformation of VSMCs. Thus, Gs-Re combined with a small amount of lysozyme may provide a novel therapeutic strategy for vascular remodeling-associated cardiovascular diseases.

Emodin Attenuates Autophagy Response to Protect the Pancreas From Acute Pancreatitis Failure.

OBJECTIVES: The aim of this study was to investigate the effects of emodin on attenuating autophagy response in acute pancreatitis (AP) models. METHODS: Acute pancreatitis was induced in Wistar rats by injecting 3% sodium taurocholate into the biliopancreatic duct. Emodin (40 mg/kg per day) was then given intragastrically, administrated 2 hours after AP induction. Rats were killed 24 hours after AP induction. The pancreatic injury was assessed using biochemical and histological approaches. Autophagosomes in pancreatic acinar cells were observed by electron microscopy. The expression levels of microtubule-associated protein 1 light chain 3 (LC3) B/A, beclin-1, and p62/SQSTM1 (p62) were detected by Western blotting, quantitative real-time polymerase chain reaction, and immunohistochemistry in pancreatic tissues. RESULTS: Compared with non-emodin-treated rats, the pathological injuries of the pancreas of emodin-treated rats were significantly alleviated, and autophagy vacuole formation was reduced within pancreatic acinar cells. Administration of emodin led to a reduction in the autophagy-associated protein level of LC3 (B/A) and p62 but not beclin-1. The transcript levels of LC3B, beclin-1, and p62 were decreased in the emodin-treated rats compared with non-emodin-treated rats. CONCLUSIONS: Our data demonstrate that emodin plays a critical role in ameliorating AP, possibly by down-regulating autophagic protein levels.

Herbal compound 861 prevents hepatic fibrosis by inhibiting the TGF-ß1/Smad/SnoN pathway in bile duct-ligated rats.

BACKGROUND: This study was to evaluate the effects of herbal compound 861 (Cpd861) on ski-related novel protein N (SnoN) and transforming growth factor-ß1 (TGF-ß1) /Smad signaling in rats with bile duct ligation (BDL)-induced hepatic fibrosis, and to explore the mechanisms of Cpd861 on hepatic fibrosis. METHODS: Thirty Wistar male rats were randomly divided into three groups: sham operation, BDL, and Cpd861. To induce hepatic fibrosis, BDL and Cpd861 group rats underwent bile duct ligation. Cpd861 at 9 g/kg/d or an equal volume of normal saline was administered intragastrically for 28 days. Liver injury was assessed biochemically and histologically. Protein and mRNA changes for SnoN and TGF-ß1/Smad signaling (TGF-ß1, Smad2, phosphorylated Smad2 [p-Smad2], phosphorylated Smad3 [p-Smad3], fibronectin, and collagen III) were determined by Western blotting and quantitative real-time PCR. RESULTS: BDL rats treated with Cpd861 had significantly alleviated hepatic fibrosis compared to BDL rats not receiving Cpd861 treatment. Moreover, Cpd861 decreased the expression of fibrosis-associated proteins fibronectin and collagen III in liver tissue. Cpd861 administration increased the expression of SnoN protein, did not change SnoN mRNA level, and decreased TGF-ß1, p-Smad2, and p-Smad3 protein expression compared to BDL without Cpd861 treatment. CONCLUSIONS: Cpd861 attenuates hepatic fibrosis by increasing SnoN protein expression and inhibiting the TGF-ß1/Smad signaling pathway.

Synergistic cytotoxicity and mechanism of caffeine and lysozyme on hepatoma cell line HepG2.

The influences of caffeine, lysozyme and the joint application of them on the hepatoma cell line HepG2 proliferation inhibition and cell apoptosis were observed by 3-(4, 5-dimethyl-2-thiazyl)-2, 5-diphenyl-2H-tetrazolium bromide assay and Hoechst 33342, which showed the proliferation inhibition rate of the joint application on HepG2 cells was 47.21%, significantly higher than caffeine or lysozyme, and the joint application promoted the apoptosis of HepG2 cells obviously. Van't Hoff classical thermodynamics formula, the Föster theory of non-radiation energy transfer and fluorescence phase diagram were used to manifest that the process of lysozyme binding to caffeine followed a two-state model, which was spontaneous at low temperature driven by enthalpy change, and the predominant intermolecular force was hydrogen bonding or Van der Waals force to stabilize caffeine-lysozyme complex with the distance 5.86nm. The attenuated total reflection-Fourier transform infrared spectra indicated that caffeine decreased the relative contents of α-helix and ß-turn, which inferred the structure of lysozyme tended to be "loose". Synchronous fluorescence spectra and ultraviolet spectra supported the above conclusion. The amino acid residues in the cleft of lysozyme were exposed and electropositivity was increased attributing to the loose structure, which were conducive to increasing caffeine concentration on the HepG2 cell surface by electrostatic interaction to show synergistic effect.

Arkadia protein expression is reduced in the liver during the progression of hepatic fibrosis.

Arkadia is able to degrade key signaling molecules in the transforming growth factor (TGF)­ß1 signaling pathway; however, the expression of Arkadia in the liver during development and progression of TGF­ß1/Smad signaling­regulated hepatic fibrosis remains to be elucidated. The present study aimed to examine Arkadia expression in the livers of two rat models of hepatic fibrosis induced by bile duct ligation and carbon tetrachloride intoxication, and in human liver samples from patients with hepatic fibrosis. Expression was analyzed by quantitative polymerase chain reaction, immunohistochemistry and western blot analysis. The results indicated that Arkadia was predominantly expressed in the cytoplasm of cholangiocytes and hepatocytes. The protein expression levels of Arkadia were significantly decreased in fibrotic livers, whereas the mRNA expression levels of Arkadia were significantly increased in fibrotic livers compared with in nonfibrotic livers. In conclusion, these data indicated that Arkadia may regulate the pathogenesis and progression of hepatic fibrosis.

Angiotensin 1-7 ameliorates caerulein-induced inflammation in pancreatic acinar cells by downregulating Toll-like receptor 4/nuclear factor-κB expression.

The present study aimed to investigate the effects of angiotensin (Ang) 1-7 on caerulein (CAE)-stimulated nuclear factor (NF)­κB, Toll­like receptor (TLR4) and cytokine expression using pancreatic acinar AR42J cells. AR42J cells were treated with 10 nmol/l CAE for various durations. In addition, cells were pretreated with various concentrations of Ang 1­7 or A779, a specific antagonist of Ang 1­7, and were stimulated with CAE for 12 h. Control cells were treated with vehicle (F­12K complete medium with 2% fetal bovine serum, 10 U/ml penicillin and 100 mg/ml streptomycin) alone. The mRNA and protein expression levels of TLR4, NF­κB, interleukin (IL)­6, IL­8, IL­10 and tumor necrosis factor­α (TNF­α) were determined by western blotting, immunofluorescence and reverse transcription­quantitative polymerase chain reaction. CAE treatment stimulated TLR4 and NF­κB expression within AR42J cells. Immunofluorescence indicated that TLR4 was expressed on the membranes and in the cytoplasm of AR42J cells, whereas NF­κB expression accumulated in the cytoplasm and nuclei. CAE­induced expression of TLR4 and NF­κB within AR42J cells was abrogated by 10­5 mmol/l Ang 1­7; however, TLR4 and NF­κB expression was enhanced with the addition of A779, particularly 10­5 mmol/l. In addition, treatment with 10­6 and 10­5 mmol/l Ang 1­7 significantly mitigated CAE­induced expression of IL­6, IL­8 and TNF­α, whereas it enhanced IL­10 expression. Conversely, A779 treatment enhanced the CAE­induced expression of IL­6, IL­8 and TNF­α, and reduced IL­10 expression in AR42J cells. In conclusion, these results suggested that Ang 1­7 may attenuate CAE­induced inflammation by downregulating TLR4, NF­κB and proinflammatory cytokine expression within AR42J cells. Therefore, Ang 1­7 may exert protective effects against the pathological progression of AP in a cell model of AP induced by CAE and may be considered in the development of treatments for this disease.

Angiotensin-converting enzyme 2-angiotensin (1-7)-Mas axis prevents pancreatic acinar cell inflammatory response via inhibition of the p38 mitogen-activated protein kinase/nuclear factor-κB pathway.

The aim of the present study was to investigate the role of the angiotensin-converting enzyme (ACE)2-angiotensin­(Ang)-(1-7)-Mas axis in the pathogenesis of pancreatitis and the association between this axis and the p38 mitogen-activated protein kinase (p38 MAPK)/nuclear factor (NF-κB) signaling pathway in pancreatic acinar cells. Mouse pancreatic acinar cancer (MPC-83) cells were stimulated with 10 nM caerulein (CAE) to create an in vitro model of acute pancreatitis, and collected for analysis at 2, 6, 12, 24 and 48 h post stimulation. In addition, cells were pretreated with different concentrations of Ang­(1­7), Ang­(1­7) antagonist A779, p38 MAPK inhibitor SB203580 or ACE2 inhibitor DX600 for 30 min, and then stimulated with CAE for 24 h. The ACE2, Mas receptor, p38 MAPK, phosphorylated (p)-p38 MAPK and NF-κB expression levels were evaluated using western blotting and immunofluorescence. p38 MAPK, NF-κB, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-8 and IL-10 mRNA expression levels were assessed using reverse transcription-quantitative polymerase chain reaction. The results of the immunofluorescence assay demonstrated that ACE2 and p38 MAPK were present mainly in the cytoplasm, while the Mas receptor was located mainly in the cell membrane. ACE2, p38 MAPK and p-p38 MAPK protein levels were significantly increased (P<0.05) following stimulation with CAE compared with those in the control group and peaked at 24 h. Mas receptor protein levels were significantly upregulated (P<0.05) between 6 and 24 h, peaking at 12 h. Ang­(1­7) and SB203580 downregulated p-p38 MAPK and NF-κB expression and the mRNA levels of inflammatory factors IL-6, TNF-α and IL-8, but upregulated the mRNA level of inflammatory factor IL-10 compared with those treated with CAE alone. These results were supported by the opposite outcomes observed for cells treated with A779 or DX600. Therefore, it was concluded that the ACE2-Ang­(1­7)-Mas axis significantly inhibits pancreatitis by inhibition of the p38 MAPK/NF-κB signaling pathway.

Angiotensin­(1­7) attenuates caerulein­induced pancreatic acinar cell apoptosis.

Extensive apoptosis of pancreatic acinar cells frequently occurs in acute pancreatitis (AP), and has been identified to be closely associated with the decrease of pancreatic parenchymal cells and pancreatic damage. The present study aimed to investigate the possible effect of angiotensin (Ang)­(1­7) on caerulein (CAE)­induced pancreatic acinar cell apoptosis. Mouse pancreatic acinar cancer cells (MPC­83) were divided into 4 groups: Control group; CAE group; CAE + Ang­(1­7) group; and CAE + Ang­(1­7) antagonist (A779) group. The control group consisted of normal MPC­83 cells without special treatment. The CAE group was stimulated with 10 nmol/l CAE and harvested at 2, 6, 12, 24 and 48 h. For the CAE + Ang­(1­7) group and CAE + A779 group, the CAE­induced pancreatic acinar cells were mock pretreated or pretreated with different concentrations of Ang­(1­7) or A779 (10­7, 10­6 or 10­5 mol/l) for 30 min. Caspase­3 is a critical executioner of apoptosis, as it is either partly or completely responsible for the proteolytic cleavage of numerous key proteins including the nuclear enzyme poly (ADP­ribose) polymerase. Activation of caspase­3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments. Thus, the present study investigated the apoptotic markers, including cleaved caspase­3, B­cell lymphoma 2 (Bcl­2), Bcl­2­like protein 4 (Bax) and renin­angiotensin system (RAS) pathway related proteins (ACE2 and Mas receptor). The results demonstrated that the cleaved caspase­3 levels were increased in the CAE group (P<0.05), peaking at 24 h, and declined when incubated with Ang­(1­7). Following treatment with Ang­(1­7), levels of the anti­apoptotic protein Bcl­2 rose dramatically in a dose­dependent manner. The ratio of the pro­apoptotic protein Bax to the anti­apoptotic protein Bcl­2 dropped notably, which demonstrated a tendency towards curbing apoptosis. In addition, the cleaved caspase­3 levels, and the ratio of Bax to Bcl­2 in the CAE + A779 group presented a significant rise compared with the CAE group. It was concluded that Ang­(1­7) may possess an inhibitory effect on CAE­induced pancreatic acinar cell apoptosis and that appropriate interventions in RAS may attenuate pancreatic injury during AP.

Fibronectin expression is critical for liver fibrogenesis in vivo and in vitro.

Increased fibronectin (FN) expression has an important role during liver fibrosis. The present study examined FN expression in rats subjected to carbon tetrachloride (CCl4)­induced liver fibrosis. In addition, the potential mechanisms underlying fibrogenesis were investigated by exposing hepatic stellate cells (HSCs) to transforming growth factor­ß (TGF­ß), which is a known inducer of myofibroblastic transformation of HSCs. Briefly, a rat model of liver fibrosis was created by administering intraperitoneal injections of CCl4. Furthermore, HSC­T6 cells were stimulated with increasing doses of recombinant TGF­ß over 24 h. Hepatic fibrosis gradually increased following CCl4 administration in vivo. Western blotting and immunohistochemistry demonstrated that fibronectin (FN), TGF­ß and α­smooth muscle actin (SMA) expression was increased following CCl4 injection, and the maximum expression levels were observed at 8 weeks. Once CCl4 treatment had been terminated, the expression levels of FN, TGF­ß and α­SMA progressively declined to near baseline levels. Western blotting and quantitative polymerase chain reaction demonstrated that FN expression was gradually increased in response to TGF­ß­stimulation of HSCs; maximum expression was achieved 12 h post­treatment (P<0.01 vs. the baseline). In conclusion, these findings indicated that FN expression is an early and progressive event that occurs during liver fibrogenesis in vivo and in vitro.

Attenuation of liver fibrosis by herbal compound 861 via upregulation of BMP-7/Smad signaling in the bile duct ligation model rat.

Herbal compound 861 (Cpd 861) exerts an anti-fibrotic effect in patients with hepatic fibrosis; however, the anti-fibrotic mechanism has yet to be fully elucidated. The present study aimed to explore the mechanistic basis for the anti-fibrotic effect, with a focus on bone morphogenetic protein 7 (BMP-7)/Smad signaling in a bile duct ligation (BDL)-induced liver fibrosis rat model. Following the induction of hepatic fibrosis, rats induced by BDL were treated with 9 g/kg Cpd 861 daily or an equal volume of saline for 28 days. Serum samples were prepared for monitoring the levels of alanine transaminase, aspartate transaminase and total bilirubin, and direct bilirubin analyses and liver samples were used to investigate gene expression, protein localization and protein expression analysis using real­time quantitative polymerase chain reaction, immunohistochemistry and western blotting. The results revealed the attenuation of liver fibrosis by Cpd 861 in the histological and biochemical experiments. BMP­7 and phospho (p)­Smad1/5/8 were localized predominantly in the cytoplasm of hepatocytes. In comparison with the saline­treated BDL rats, Cpd 861 markedly upregulated the gene expression of BMP­7 and Smad5, as well as the protein expression of BMP­7 and Smad1/5. In addition, p-Smad1/5/8 protein expression was markedly increased by Cpd 861 in the BDL model. These results indicated that Cpd 861 alleviates hepatic fibrosis possibly via the upregulation and activation of BMP-7/Smad signaling in hepatic fibrotic rats.

[Research on Molecular Spectra of Interactions Between Salvianolic Acid A and Salvianolic Acid B with Insulin, and Effect of Glucose on the Binding].

The interactions of Salvianolic acis A (SAA) and Salvianolic acid B (SAB) with insulin were studied by using fluorescence spectroscopy, UV-vis spectroscopy and ATR-FTIR spectroscopy in simulating physiological condition (pH 7.40). The fluorescence quenching of insulin by SAA and SAB were static quenching process. The results of synchronous fluorescence and three-dimensional fluorescence spectra suggested no obvious conformation changes of insulin after SAA or SAB binding. But ATR-FTIR spectra showed that SAA and SAB could change the secondary structures of insulin, of which ß-turns decreased and random coil increased accompanied with α-belices and ß-sheets no clear change. The glucose might influenced the bioactivity of insulin in the SAA-insulin and SAB-insulin systems by changing the binding constants of SAA (or SAB) with insulin and exacerbating the changes of insulin conformation and relative contents of α-belices.

The ACE2-angiotensin-(1-7)-Mas axis protects against pancreatic cell damage in cell culture.

OBJECTIVE: Angiotensin-converting enzyme 2 (ACE2), its product angiotensin-(1-7), and its receptor Mas have been shown to moderate the adverse effects of the ACE-angiotensin II-AT1 axis in many diseases. The aim of this study was to determine whether the ACE2-Ang-(1-7)-Mas axis could have similar effects in a cell culture model of pancreatic damage. METHODS: AR42J cells were stimulated with 10 nmol/L cerulein to simulate acute pancreatitis. ACE2, Ang-(1-7), Mas receptor, and PI3K/AKT pathway were measured by quantitative real-time polymerase chain reaction and Western blot analysis. RESULTS: ACE2 and Mas receptor protein levels in AR42J cells were significantly increased (P < 0.05) between 30 minutes and 6 hours postdisease induction compared with the control group. Mas receptor gene expression was significantly increased (P < 0.05) at 2 hours postdisease induction, and Ang-(1-7) was increased at 6 hours. Treatment with Ang-(1-7) in AR42J cells increased IL-10, decreased IL-6 and IL-8, and reduced the damage to pancreatic cells. Levels of IL-6 and IL-8 in AR42J cell culture were increased significantly after treatment with A779. Moreover, Ang-(1-7) increased the concentration of PI3K/AKT pathway and eNOSin AR42J cells. CONCLUSIONS: ACE2-angiotensin-(1-7)-Mas axis significantly inhibits pancreatitis in response to decreased inflammatory factors by the activation of endothelial nitric oxide synthase and NO signaling pathways.

Ulinastatin activates the renin-angiotensin system to ameliorate the pathophysiology of severe acute pancreatitis.

BACKGROUND AND AIM: Ulinastatin is a drug used effectively to alleviate symptoms and improve the pathophysiology of various types of pancreatitis. However, the molecular mechanism responsible for its action remains unknown. Therefore, we further explore the therapeutic effects of ulinastatin and investigate possible molecular pathways modulated by this drug in the development of severe acute pancreatitis (SAP). METHODS: SAP mouse model was created by administering intraperitoneal injections of cerulein and lipopolysaccharide. Pancreatic injury was assessed by performing biochemical and histological assays and by measuring the inflammatory response of the pancreas. Specifically, we examined changes in the expression of components of the rennin-angiotensin system (RAS), including angiotensin-converting enzyme (ACE)-angiotensin II (Ang II)-angiotensin type 1 receptor (AT-1R), and ACE2-Ang-(1-7)-Mas receptor. RESULTS: When SAP mouse models were treated with ulinastatin at a dosage of 50,000 U/kg body weight, we found, through biochemical and histopathological analyses, that the pancreatic injury was significantly ameliorated. Administration of ulinastatin to SAP mice led to increased expression of ACE2, Ang-(1-7), and Mas receptor, decreased expression of serum Ang II and pancreatic AT-1R, and no alterations in the expression of pancreatic ACE and Ang II when compared to cerulein-treated control group that did not receive ulinastatin. CONCLUSIONS: This study shows that ulinastatin has differential effects on the two axes of the RAS during SAP. Our results further suggest that upregulation of components of the ACE2-Ang-(1-7)-Mas pathway might be an important mechanism contributing to the therapeutic role of ulinastatin in alleviating pancreatitis-associated symptoms.

Angiotensin-converting enzyme (ACE and ACE2) imbalance correlates with the severity of cerulein-induced acute pancreatitis in mice.

Angiotensin-converting enzyme (ACE) and its effector peptide angiotensin II (Ang II) have been implicated in the pathogenesis of pancreatitis. Angiotensin-converting enzyme 2 (ACE2) degrades Ang II to angiotensin-(1-7) [Ang-(1-7)] and has recently been described to have an antagonistic effect on ACE signalling. However, the specific underlying role of ACE2 in the pathogenesis of severe acute pancreatitis (SAP) is unclear. In the present study, the local imbalance of ACE and ACE2, as well as Ang II and Ang-(1-7) expression, was compared in wild-type (WT) and ACE2 knock-out (KO) or ACE2 transgenic (TG) mice subjected to cerulein-induced SAP. Serum amylase, tumour necrosis factor-α, interleukin (IL)-1ß, IL-6 and IL-10 levels and histological morphometry were used to determine the severity of pancreatitis. In WT mice, pancreatic ACE and Ang II and serum Ang II expression increased (P < 0.05), while pancreatic ACE2 and Ang-(1-7) and serum Ang-(1-7) levels were also significantly elevated (P < 0.05) from 2 to 72 h after the onset of SAP. However, the ratio of pancreatic ACE2 to ACE expression was significantly reduced (from 1.46 ± 0.09 to 0.27 ± 0.05, P < 0.001) and paralleled the severity of pancreatitis. The Ace2 KO mice exhibited increased levels of tumour necrosis factor-α, IL-1ß, IL-6, multifocal coagulative necrosis and inflammatory infiltrate, and lower levels of serum IL-10 and pancreatic Ang-(1-7) (4.70 ± 2.13 versus 10.87 ± 2.51, P < 0.001) compared with cerulein-treated WT mice at the same time point. Conversely, Ace2 TG mice with normal ACE expression were more resistant to SAP challenge as evidenced by a decreased inflammatory response, attenuated pathological changes and increased survival rates. These data suggest that the ACE2-ACE imbalance plays an important role in the pathogenesis of SAP and that pancreatic ACE2 is an important factor in determining the severity of SAP.

Comparative studies on interactions of baicalein, baicalin and scutellarin with lysozyme.

The interactions of baicalein, baicalin and scutellarin with lysozyme (LYSO) were studied by fluorescence and UV spectroscopy. The results showed that all the three flavones can quench the fluorescence of LYSO via static quenching with the distance between the donor and acceptor less than 7 nm. The hydroxyl at B-ring gave flavones an advantage to binding with LYSO. Electrostatic forces played a major role in stabilizing baicalein-LYSO complex and baicalin-LYSO complex, whereas hydrophobic interactions in scutellarin-LYSO. Furthermore, the presence of pantothenic acid can increase the binding constant and the number of binding sites between flavones and LYSO.

[Quantitative analysis of steroidal saponins in Chinese material medica Rhizoma Paridis by HPLC-ELSD].

OBJECTIVE: The several species of the genus Paris called as Rhizoma Paridis were famous traditional Chinese medica. To develop the quantitative analysis method of the steroidal saponins in some species of the genus Paris and commercially available Rhizoma Paridis samples by HPLC-ELSD. METHOD: The contents of 11 steroidal saponins in Rhizoma Paridis samples were dectected with a Kromasil C18(4.6 mm x 150 mm, 5 microm) column which was deluted with acetonitrile-water (30:70-60:40) at a flow rate of 1 mL x min(-1) by HPLC-ELSD. RESULT: All the authentic samples could be separated and calibration curves of 11 saponins were prepared. 11 steroidal saponins in 16 Rhizoma Paridis samples were detected in 30 min. The recovery for the assay of saponins was between 95% and 97%. The precision and stability of samples (RSD) were below 3%. CONCLUSION: The method was shown to be accurate and convenient, and suitable for the quantitative analysis of these 11 steroidal saponins in the commercially available Rhizoma Paridis samples.

[Separation and identification of active constituents of Paris vietnamensis].

AIM: To study the anti-tumor bioactive steroid saponins of Paris vietnamensis (Takht.). METHODS: The constituents were isolated and purified by chromatography and their structures were identified by spectral analysis and physicochemical properties. The cytotoxic bioactivities of the constituents were determined by MTT. RESULTS: Eleven compounds were obtained and identified as 3beta, 5alpha,6alpha-trihydroxy-7(8)-en-isospirostanol-3-O-beta-D-glucopyranosyl(1-->3) [alpha-L-rhamnopyranosyl(1-->2)]-beta-D-glucopyranoside (1), which was named as parisvietnaside A, 25 (R) diosgenin-3-O-alpha-L-arabinofuranosyl(1-->4)-beta-D-glucopyranoside (2), 25(R) diosgenin-3-O-alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranoside (3), 25 (R) diosgenin-3-O-alpha-L-arabinofuranosyl (1-->4) [alpha-L-rhamnopyranosyl (1-->2)]-beta-D-glucopyranoside (4), 25 (R) diosgenin-3-O-beta-D-glucopyranosyl (1-->3) [alpha-L-rhamnopyranosyl (1-->2)]-beta-D-glucopyranoside (5), 25 (R) diosgenin-3-O-alpha-L-rhamnopyranosyl (1-->4)-alpha-L-rhamnopyranosyl(1-->4) [alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside (6), 25 (R) pennogenin-3-O-alpha-L-arabinofuranosyl(1-->4)-beta-D-glucopyranoside (7), 25 (R) pennogenin-3-O-alpha-L-rhamnopyranosyl (1-->2)-beta-D-glucopyranoside (8), 25 (R) pennogenin-3-O-alpha-L-arabinofuranosyl (1-->4) [alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside (9), 25 (R) pennogenin-3-O-beta-D-glycopyranosyl (1-->3) [alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranoside (10) and 25 (R) pennogenin-3-O-alpha-L-rhamnopyranosyl (1-->4)-alpha-L-rhamnopyranosyl(1-->4) [alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside (11). Some constituents had cytotoxic bioactivities. CONCLUSION: Compound 1 is a new spirostanol saponin, and compounds 2, 3, 6-11 were obtained from Paris vietnamensis (Takht.) for the first time. Compounds 3, 4, 6, 8 had cytotoxic bioactivities against tumor cells HepG2 and SGC-7901.