Study on the technology of efficient extraction of eleutheroside E from Acanthopanax senticosus by green solvent DES.

OBJECTIVES: Acanthopanax senticosus (Rupr. & Maxim.) Harms is a medicinal and edible plant which is clinically used for the recovery and treatment of cardiovascular and central diseases. As a characteristic active pharmaceutical ingredient of Acanthopanax senticosus, eleutheroside E is the core of the therapeutic effect. Organic solvent extraction has low selectivity, low extraction rate, difficulty in separation and purification and safety risks. The purpose of this study was to extract the effective component of Acanthopanax senticosus with a new green solvent. METHODS: In this article, two kinds of deep eutectic solvents (DESs) (DES-1 and DES-2) were synthesised by heating and stirring methods. Eleutheroside E was extracted by ultrasonic extraction with two kinds of DES as extractants and quantitatively analysed by Orbitrap-tandem mass spectrometry (MS/MS). RESULTS: The main results showed that the initial polarity of the DES was similar to that of 60 to 80% ethanol and hydrogen bond donors were the main factors affecting the polarity of DES. In the test, the viscosity of DES was higher than that of ethanol, and even the addition of a small amount of water (10%) caused intermolecular hydrogen bond disruption and redistribution of the solvent, resulting in a significant decrease in solvent viscosity. The solvents in the test group were stable after standing at 5°C in the dark for 100 days. The extraction rate of eleutheroside E by DES solvent was 5-6 times higher than that by ethanol. DES-1 and DES-2 can efficiently extract eleutheroside E. CONCLUSION: This study developed a new method for the application of the green extraction of eleutheroside E with certain practical significance.

Diagnostic Performance of FibroTouch Ultrasound Attenuation Parameter and Liver Stiffness Measurement in Assessing Hepatic Steatosis and Fibrosis in Patients With Nonalcoholic Fatty Liver Disease.

INTRODUCTION: To evaluate the diagnostic performance of ultrasound attenuation parameter (UAP) and liver stiffness measurement (LSM) by FibroTouch for diagnosis of hepatic steatosis and fibrosis in patients with nonalcoholic fatty liver disease (NAFLD). METHODS: We recruited 237 patients undergoing FibroTouch and liver biopsy within 2 weeks. The pathological findings of liver biopsy were scored by Nonalcoholic Steatohepatitis Clinical Research Network, and the diagnostic accuracy of UAP for steatosis and LSM for fibrosis was evaluated by area under the receiver operating characteristic curve (AUROC). The impacts of histological parameters on UAP and LSM were analyzed, and diagnostic performance of FibroTouch UAP and LSM was compared with other noninvasive biomarkers. RESULTS: The success rate of FibroTouch examination was 96.51%. The AUROC of UAP for diagnosis of steatosis ≥S1, ≥S2, and S3 was 0.88, 0.93, and 0.88, and the cutoff values were 244, 269, and 296 dB/m, respectively. The AUROC of LSM for the diagnosis of fibrosis stages ≥F2, ≥F3, and F4 was 0.71, 0.71, and 0.77, and the cutoff values were 9.4, 9.4, and 11 kPa, respectively. Multiple regression analysis showed that LSM was positively correlated with degree of fibrosis and NAFLD activity score. UAP was positively correlated with liver steatosis. The diagnostic performance of UAP for steatosis was significantly superior to that of the hepatic steatosis index. DISCUSSION: FibroTouch has a low failure rate with moderate to high diagnostic performance for discriminating the steatosis degree and fibrosis stage and is suitable for clinical evaluation and monitoring of patients with NAFLD.

High Incidence of Escherichia coli Strains Coharboring mcr-1 and blaNDM from Chickens.

This study investigated the characteristics of Escherichia coli isolates carrying mcr-1-blaNDM from a chicken farm in China. Of the 78 E. coli isolates, 21 clonally unrelated isolates carried mcr-1-blaNDM Diverse IncI2 plasmids disseminated mcr-1, while the dissemination of blaNDM was mediated by diverse IncB/O plasmids. More striking was the colocalization of resistance genes mcr-1 and blaNDM-4 in an IncHI2/ST3 plasmid, which might pose a great challenge for public health.

Efficacy and safety of gemcitabine-based chemotherapies in biliary tract cancer: a meta-analysis.

AIM: To investigate the efficacy and safety of gemcitabine (Gem)-based combination chemotherapies for the treatment of advanced biliary tract cancer. METHODS: Clinical trials were identified by searching scientific literature databases (PubMed, EMBASE and the Cochrane Library) for studies published between 1975 and 2013. Two reviewers independently evaluated the relevant studies and manually searched references from these reports to locate additional eligible studies. The disease response and control rates, progression-free and overall survivals, and the grade 3-4 toxicities were evaluated by a meta-analysis. Odds-ratios (ORs) of the disease response and control rates and grade 3-4 toxicities, and the mean difference (MD) of both progression-free and overall survivals were calculated and used for statistical analysis. RESULTS: Seven randomized trials with a total of 858 patients were selected and included in the final analysis. The studies were divided into subgroups based on the chemotherapy regimens, including Gem-based and non-Gem-based chemotherapies. The overall analyses revealed that the patients treated with Gem-based combination chemotherapy had significantly higher disease response rates [OR = 1.69, 95% confidence interval (CI): 1.17-2.43; P = 0.01], a longer progression-free survival (MD = 1.95, 95%CI: 0.90-3.00; P = 0.00) and a longer overall survival (MD = 1.85, 95%CI: 0.26-3.44; P = 0.02). A higher incidence of grade 3-4 hematological toxicities, including leukopenia (OR = 2.98, 95%CI: 1.44-6.20; P = 0.00), anemia (OR = 2.96, 95%CI: 1.79-4.92; P = 0.00) and neutropenia (OR = 2.80, 95%CI: 1.39-5.64; P = 0.00) was found in the Gem-based combination chemotherapy group compared with the Gem monotherapy and non-Gem-based chemotherapy groups. CONCLUSION: Gem-based combination chemotherapy is a potential first-line treatment for advanced biliary tract cancer as a result of improved survival, though with additional toxicity.

ROCK is involved in vasculogenic mimicry formation in hepatocellular carcinoma cell line.

Ras homolog family member A (RhoA) and Rho-associated coiled coil-containing protein kinases 1 and 2 (ROCK1 and 2) are key regulators of focal adhesion, actomyosin contraction and cell motility. RhoA/ROCK signaling has emerged as an attractive target for the development of new cancer therapeutics. Whether RhoA/ROCK is involved in regulating the formation of tumor cell vasculogenic mimicry (VM) is largely unknown. To confirm this hypothesis, we performed in vitro experiments using hepatocellular carcinoma (HCC) cell lines. Firstly, we demonstrated that HCC cells with higher active RhoA/ROCK expression were prone to form VM channels, as compared with RhoA/ROCK low-expressing cells. Furthermore, Y27632 (a specific inhibitor of ROCK) rather than exoenzyme C3 (a specific inhibitor of RhoA) effectively inhibited the formation of tubular network structures in a dose-dependent manner. To elucidate the possible mechanism of ROCK on VM formation, real-time qPCR, western blot and immunofluorescence were used to detect changes of the key VM-related factors, including VE-cadherin, erythropoietin-producing hepatocellular carcinoma-A2 (EphA2), phosphoinositide 3-kinase (PI3K), matrix metalloproteinase (MMP)14, MMP2, MMP9 and laminin 5γ2-chain (LAMC2), and epithelial-mesenchymal-transition (EMT) markers: E-cadherin and Vimentin. The results showed that all the expression profiles were attenuated by blockage of ROCK. In addition, in vitro cell migration and invasion assays showed that Y27632 inhibited the migration and invasion capacity of HCC cell lines in a dose-dependent manner markedly. These data indicate that ROCK is an important mediator in the formation of tumor cell VM, and suggest that ROCK inhibition may prove useful in the treatment of VM in HCC.

TET3 mediates the activation of human hepatic stellate cells via modulating the expression of long non-coding RNA HIF1A-AS1.

Activated Hepatic stellate cells (HSCs) play a critical role in liver fibrosis and a lot of efforts have been made to dissect the underlying mechanism involved in activation of HSCs. However, the underlying mechanism remains douteux up to now. In the present study, we found that TET3, one member of ten-eleven translocation (TET) protein family, reduced significantly in HSCs LX-2 activated by TGF-ß1. To study the function of TET3 in activation of HSCs, knockdown was performed by RNA interference. Results showed that cell proliferation rise significantly and cell apoptosis reduce obviously after knockdown of TET3. Meanwhile, IHC showed that the expression of α-SMA rise significantly compared to control. These results indicated that TET3 is closely associated with the activation of HSCs. Further studies found that long non-coding RNA HIF1A-AS1 was reduced significantly in LX-2 cell after treatment with siRNA for TET3. The result hinted that TET3 activate HSCs through modulating the expression of HIF1A-AS1. To confirm this hypothesis, RNA interference was performed to silence the HIF1A-AS1. Results showed that HIF1A-AS1 silencing lead to enhancing in cell proliferation and declining apoptosis. Taken together, TET3 can mediate the activation of HSCs via modulating the expression of the long non-coding RNA HIF1A-AS1.

Residue depletion of nifuroxazide in broiler chicken.

BACKGROUND: Several nitrofuran drugs have been prohibited for use in food producing animals due to their carcinogenic and mutagenic effects. However, one of the nitrofurans, nifuroxazide, is still used as a veterinary drug in some countries. This study was conducted to investigate the residue depletion of nifuroxazide in broiler chicken. Chickens were fed with dietary feeds containing 50 mg kg⁻¹ of nifuroxazide for seven consecutive days. Liver, kidney, muscle and plasma samples were collected at different withdrawal periods, and the residues of parent nifuroxazide and its acid-hydrolysable side chain, 4-hydroxybenzhydrazide (HBH), in these samples were determined. RESULTS: Nifuroxazide was metabolised in vivo and its metabolite HBH was formed. Parent nifuroxazide was not detectable in these samples after 14 days of cessation. HBH was detectable in these samples even after 28 days of cessation and the total HBH residues were higher than 1.0 ng g⁻¹. Furthermore, the residue level of tissue bound HBH was much higher than that of free HBH. CONCLUSION: The tissue-bound HBH could be used as a marker to monitor the residue of nifuroxazide in chicken and the best target tissue should be liver. This is the first paper reporting the residue depletion of nifuroxazide in chicken.