Mechanism of lncRNA SNHG16 on kidney clear cell carcinoma cells by targeting miR-506-3p/ETS1/RAS/ERK molecular axis.

Objective: This study aimed to investigate the mechanism of long noncoding ribonucleic acid (lncRNA) SNHG16 on kidney clear cell carcinoma (KIRC) cells by targeting miR-506-3p/ETS proto-oncogene 1, transcription factor (ETS1)/RAS/Extracellular regulated protein kinases (ERK) molecular axis, thus to provide reference for clinical diagnosis and treatment of KIRC in the future. Methods: Thirty-six patients with KIRC were enrolled in this study, and their carcinoma tissues and adjacent tissues were obtained for the detection of SNHG16/miR-506-3p/ETS1/RAS/ERK expression. Then, over-expressed SNHG16 plasmid and silenced plasmid were transfected into KIRC cells to observe the changes of their biological behavior. Results: SNHG16 and ETS1 were highly expressed while miR-506- 3p was low expressed in KIRC tissues; the RAS/ERK signaling pathway was significantly activated in KIRC tissues (P < 0.05). After SNHG16 silence, KIRC cells showed decreased proliferation, invasion and migration capabilities and increased apoptosis rate; correspondingly, increase in SNHG16 expression achieved opposite results (P < 0.05). Finally, in the rescue experiment, the effects of elevated SNHG16 on KIRC cells were reversed by simultaneous increase in miR-506-3p, and the effects of miR-506-3p were reversed by ETS1. Activation of the RAS/ERK pathway had the same effect as increase in ETS1, which further worsened the malignancy of KIRC. After miR-506-3p increase and ETS1 silence, the RAS/ERK signaling pathway was inhibited (P < 0.05). At last, the rescue experiment (co-transfection) confirmed that the effect of SNHG16 on KIRC cells is achieved via the miR-506-3p/ETS1/RAS/ERK molecular axis. Conclusion: SNHG16 regulates the biological behavior of KIRC cells by targeting the miR-506-3p/ETS1/RAS/ERK molecular axis.

Pharmacokinetics of ceftiofur sodium in cats following a single intravenous and subcutaneous injection.

Ceftiofur, a third-generation cephalosporin antibiotic, is being extensively used by pet doctors in China. In the current study, the detection method was developed for ceftiofur and its metabolites, desfuroylceftiofur (DCE) and desfuroylceftiofur conjugates (DCEC), in feline plasma. Then, the pharmacokinetics studies were performed following one single intravenous and subcutaneous injection of ceftiofur sodium in cats both at 5 mg/kg body weight (BW) (calculated as pure ceftiofur). Ceftiofur, DCE, and DCEC were extracted from plasma samples, then derivatized and further quantified by high-performance liquid chromatography. The concentrations versus time data were subjected to noncompartmental analysis to obtain the pharmacokinetics parameters. The terminal half-life (t1/2λz ) was calculated as 11.29 ± 1.09 and 10.69 ± 1.31 hr following intravenous and subcutaneous injections, respectively. After intravenous treatment, the total body clearance (Cl) and volume of distribution at steady-state (VSS ) were determined as 14.14 ± 1.09 ml hr-1  kg-1 and 241.71 ± 22.40 ml/kg, respectively. After subcutaneous injection, the peak concentration (Cmax ; 14.99 ± 2.29 µg/ml) was observed at 4.17 ± 0.41 hr, and the absorption half-life (t1/2ka ) and absolute bioavailability (F) were calculated as 2.83 ± 0.46 hr and 82.95%±9.59%, respectively. The pharmacokinetic profiles of ceftiofur sodium and its related metabolites demonstrated their relatively slow, however, good absorption after subcutaneous administration, poor distribution, and slow elimination in cats. Based on the time of drug concentration above the minimum inhibitory concentration (MIC) (T>MIC) calculated in the current study, an intravenous or subcutaneous dose at 5 mg/kg BW of ceftiofur sodium once daily is predicted to be effective for treating feline bacteria with a MIC value of ≤4.0 µg/ml.

Alisertib, an Aurora kinase A inhibitor, induces apoptosis and autophagy but inhibits epithelial to mesenchymal transition in human epithelial ovarian cancer cells.

Ovarian cancer is a leading killer of women, and no cure for advanced ovarian cancer is available. Alisertib (ALS), a selective Aurora kinase A (AURKA) inhibitor, has shown potent anticancer effects, and is under clinical investigation for the treatment of advanced solid tumor and hematologic malignancies. However, the role of ALS in the treatment of ovarian cancer remains unclear. This study investigated the effects of ALS on cell growth, apoptosis, autophagy, and epithelial to mesenchymal transition (EMT), and the underlying mechanisms in human epithelial ovarian cancer SKOV3 and OVCAR4 cells. Our docking study showed that ALS, MLN8054, and VX-680 preferentially bound to AURKA over AURKB via hydrogen bond formation, charge interaction, and π-π stacking. ALS had potent growth-inhibitory, proapoptotic, proautophagic, and EMT-inhibitory effects on SKOV3 and OVCAR4 cells. ALS arrested SKOV3 and OVCAR4 cells in G2/M phase and induced mitochondria-mediated apoptosis and autophagy in both SKOV3 and OVCAR4 cell lines in a concentration-dependent manner. ALS suppressed phosphatidylinositol 3-kinase/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) and p38 mitogen-activated protein kinase pathways but activated 5'-AMP-dependent kinase, as indicated by their altered phosphorylation, contributing to the proautophagic activity of ALS. Modulation of autophagy altered basal and ALS-induced apoptosis in SKOV3 and OVCAR4 cells. Further, ALS suppressed the EMT-like phenotype in both cell lines by restoring the balance between E-cadherin and N-cadherin. ALS downregulated sirtuin 1 and pre-B cell colony enhancing factor (PBEF/visfatin) expression levels and inhibited phosphorylation of AURKA in both cell lines. These findings indicate that ALS blocks the cell cycle by G2/M phase arrest and promotes cellular apoptosis and autophagy, but inhibits EMT via phosphatidylinositol 3-kinase/Akt/mTOR-mediated and sirtuin 1-mediated pathways in human epithelial ovarian cancer cells. Further studies are warranted to validate the efficacy and safety of ALS in the treatment of ovarian cancer.

Mussel-inspired adhesive and transferable free-standing films by self-assembling dexamethasone encapsulated BSA nanoparticles and vancomycin immobilized oxidized alginate.

This study developed an adhesive and transferable free-standing (FS) film with dual function of osteoinductivity and antibacterial activity, which was obtained by sequentially assembling vancomycin immobilized oxidized sodium alginate and dexamethasone encapsulated chitosan coated BSA nanoparticles on a poly-dopamine layer. The FS films enabled the dual release of vancomycin and dexamethasone. The FS films had excellent osteoinductivity and antibacterial activity by cell culture and antibacterial assay. The FS film was detached from substrates and transferred to non-fouling surfaces by a wet transfer method, which demonstrated that the adhesive FS film is potential to modify biopolymers with non-fouling surfaces in mild and biocompatible conditions for biomedical applications.

[Research on relevance between macroscopic feature individual differences and quality of Gansu cultivated Angelica sinensis].

OBJECTIVE: To analyze the appearance differences of Gansu cultivated Angelica sinensis, and explore the relevance between the appearance differences and quality. METHODS: The macroscopic feature of 22 batches of Angelica sinensis from different habitats was measured as index. The content of ferulic acid, volatile oil and extract were determined by the method recorded in the Chinese Pharmacopoeia. The data were analyzed by SPSS 15.0 software. RESULTS: The habitat was positively correlated with the index. Indexes of nine groups had direct correlation with each other. The habitat was significantly correlated with other indexes except the length of the head. The extract and ferulic acid were positively correlated with habitat and index. Extract had significant correlation with macroscopic feature. Ferulic acid only had significant correlation with head length. The volatile oil only had significant correlation with habitat and no significant correlation with index. Root weight and number of lateral roots had obvious difference in different habitat which coefficient of variation was 44.1% and 28.6%, respectively. CONCLUSION: There are significant individual differences in Angelica sinensis. The chemical composition has a certain correlation with macroscopic feature. Angelica sinensis cultivation needs to consider the choice of habitat.

Effect of Fructus Psoraleae on motility of gallbladder isolated smooth muscle strips from guinea pigs.

AIM: To observe the effect of Fructus Psoraleae on motility of isolated gallbladder muscle strips of guinea pigs and its mechanism. METHODS: Guinea pigs were hit to lose consciousness and the whole gallbladder was removed quickly. Two or three smooth muscle strips (8 mm x 3 mm) were cut along a longitudinal direction. The mucosa was gently removed. Every longitudinal muscle strip was suspended in a tissue chamber which was continuously perfused with 5 mL Krebs solution (37 degrees of C), pH 7.4, and aerated with 950 mL/L O2 and 50 mL/L CO2. The isometric response was recorded with an ink-writing recorder. After 2 h equilibration under 1 g-load, 50 microL Fructus Psoraleae (10, 20, 70, 200, 700, 1000 g/L) was added cumulatively into the tissue chamber in turn every 2 min to observe their effects on gallbladder muscle strips (cumulating final concentration of Fructus Psoraleae was 0.1, 0.3, 1.0, 3.0, 10.0, 20.0 g/L). The antagonists, including 4-DAMP, benzhydramine, hexamethonium, phentolamine, verapamil and idomethine were given 2 min before Fructus Psoraleae respectively to investigate the mechanisms involved. RESULTS: Fructus Psoraleae dose-dependently increased the resting tension (r = 0.992, P < 0.001), decreased the mean contractile amplitude (r = 0.970, P < 0.001) and meanwhile increased the contractile frequency of the gallbladder muscle strip in vitro (r = 0.965, P < 0.001). The exciting action of Fructus Psoraleae on the resting tension could be partially blocked by 4-DAMP (the resting tension decreased from 1.37 +/- 0.41 to 0.70 +/- 0.35, P < 0.001), benzhydramine (from 1.37 +/- 0.41 to 0.45 +/- 0.38, P < 0.001), hexamethonium (from 1.37 +/- 0.41 to 0.94 +/- 0.23, P < 0.05), phentolamine ( from 1.37 +/- 0.41 to 0.89 +/- 0.22, P < 0.01) and verapamil (from 1.37 +/- 0.41 to 0.94 +/- 0.26, P < 0.05). But the above antagonists had no significant effect on the action of Fructus Psoraleae-induced mean contractile amplitude (P > 0.05). Moreover, the increase of the contractile frequency due to Fructus Psoraleae was inhibited by 4-DAMP (decreased from 8.3 +/- 1.2 to 6.8 +/- 0.5, P < 0.01) and hexamethonium (from 8.3 +/- 1.2 to 7.0 +/- 0.9, P < 0.05). Idomethine had no significant effect on the Fructus Psoraleae-induced responses (P > 0.05). CONCLUSION: Fructus Psoraleae enhances the motility of isolated gallbladder muscle strips from guinea pigs, in a dose-dependent manner. The effect of Fructus Psoraleae is partly related to M3, N receptor, alpha receptor, H1 receptor, Ca2+ channel, but not related to prostaglandin.

Effects of rhubarb on isolated gastric muscle strips of guinea pigs.

AIM: To study the effects of rhubarb (dried root of Rheum officinale Baill.) on contractile activity of isolated gastric muscle strips of guinea pigs and its possible mechanism. METHODS: A total of 48 guinea pigs were killed to remove the whole stomach. Then, the stomach was opened and the mucosal layer was removed. Parallel to the circular fibers, muscle strips were cut from the body. Each isolated gastric muscle strip was suspended in a tissue chamber containing 5 mL Krebs solution, constantly warmed by water jacket at 37 degrees and bubbled continuously with a mixed gas of 950 mL/L O2 and 50 mL/L CO2. After being incubated for 1 h with 1 g tension, rhubarb of varied concentrations (1%, 2%, 7%, 20% and 70%) was added cumulatively into the tissue chamber at intervals of 2 min. Atropine (10(-6) mol/L) or isoptin (5 x 10(-8) mol/L) or hexamethonium (10(-5) mol/L) was given 2 min before the administration of rhubarb. The isometrical response was measured with an ink-writing recorder. RESULTS: Rhubarb dose dependently increased the resting tension of gastric body circular muscle (CM) (r = 0.726, P<0.05). Atropine (r = 0.829, P<0.05), isoptin (r = 0.764, P<0.05) and hexamethonium (r = 0.797, P<0.05) did not affect its action in a dose-related manner. Atropine apparently reduced the increasing action of 1%, 3%, 10%, 30% and 100% rhubarb on the resting tension of gastric body CM. Isoptin inhibited the effect of 10%, 30% and 100% rhubarb on the resting tension of gastric body CM. Hexamethonium reduced the increasing action of 1%, 10%, 30% and 100% rhubarb on the resting tension of gastric body CM. Rhubarb increased the contractile frequency of CM of body. While atropine, isoptin and hexamethonium did not inhibit the contractile frequency of gastric body CM in comparison with rhubarb at the same concentration, rhubarb at the highest concentration (100%) decreased the mean contractile amplitude of gastric body CM. Atropine, isoptin and hexamethonium did not affect the mean contractile amplitude of gastric body CM compared to rhubarb at the same concentration. CONCLUSION: Rhubarb has exciting actions on isolated gastric smooth muscle strips of guinea pig. The exciting action of rhubarb is partly mediated via cholinergic M receptor, cholinergic N receptor and L-type calcium channel.

Effect of areca on contraction of colonic muscle strips in rats.

AIM: To investigate the effects of areca on the contractile activity of isolated colonic muscle strips in rats and mechanism involved. METHODS: Each strip (LMPC, longitudinal muscle of proximal colon; CMPC, circular muscle of proximal colon; LMDC, longitudinal muscle of distal colon; CMDC, circular muscle of distal colon.) was suspended in a tissue chamber containing 5 mL Krebs solution (37 degrees C), bubbled continuously with 950 mL.L(-1) O(2) and 50 mL.L(-1) CO(2). The mean contractile amplitude (A), the resting tension (T), and the contractile frequency (F) were simultaneously recorded on recorders. RESULTS: Areca dose dependently increased the mean contractile amplitude, the resting tension of proximal and distal colonic smooth muscle strips in rats (P<0.05). It also partly increased the contractile frequency of colonic smooth muscle strips in rats (P<0.05). The effects were partly inhibited by atropine (the resting tension of LMPC decreased from 0.44 +/- 0.12 to 0.17 +/- 0.03; the resting tension of LMDC decreased from 0.71 +/- 0.14 to 0.03 +/- 0.01; the mean contractile amplitude of LMPC increased from -45.8 +/- 7.2 to -30.5 +/- 2.9; the motility index of CMDC decreased from 86.6 +/- 17.3 to 32.8 +/- 9.3; P<0.05 vs areca), but the effects were not inhibited by hexamethonium (P>0.05). CONCLUSION: Areca stimulated the motility of isolated colonic smooth muscle strips in rats. The stimulation of areca might be relevant with M receptor partly.

Effects of Dangshen on isolated gastric muscle strips in rats.

AIM:To study the effects of Dangshen dried root of Codonopsis Pilosula (Franch) Nannf on contractile activity of isolated gastric muscle strips in rats and its possible mechanism involved.METHODS:Each isolated gastric muscle strip was put in a tissue chamber containing 5ml Krebs solution, constantly warmed by water jacket at 37?mgr; and supplied with a mixed gas of 95% O(2) and 5% CO(2). After incubating for 1h with 1g tension, Dangshen of varied concentration was added cumulatively in the tissue chamber at intervals of 2 minutes. The isometrical response was measured on ink-writing recorders.RESULTS:Dangshen dose dependence increased the resting tension of longitudinal muscle (LM) of fundus (r =0.96, P < 0.01), the mean contractile amplitude of circular muscle (CM) of the stomach body (r =0.87, P < 0.05) and CM of antrum (r =0.98, P < 0.01), and the motility index CM of pylorus(r =0.87, P < 0.05). Atropine (5 10( 8)mol/L) or Hexamethonium (10( 5)mol/L) or Indomethacin (5 10( 7)mol/L) was given 2 minutes before the administration of Dangshen, it did not abolish its dose related manner. Atropine apparently reduced the increasing action of 10% and 30% Dangshen on the resting tesion of LM of fundus (P < 0.05), 30%, 100% and 200% Dangshen on bodied strips (P < 0.05), 100% and 200% Dangshen on antral strips (P < 0.05).Hexamethonium reduced the increasing action of 10% and 30% Dangshen on the resting tesion of LM of fundus (P < 0.05 and P < 0.05), 30%, 100% and 200% Dangshen on bodied strips (P < 0.05), and 100% and 200% Dangshen on pyloric strips (P < 0.05). Indomethacin inhibited the effect of 10% Dangshen on the resting tesion of LM of fundus (P < 0.05), but did not affect the exciting action of Dangshen on strips of body, antrum and pylorus.CONCLUSION:The results showed that Dangshen possessed exciting action on the isolated gastric smooth muscle strips of the rat.The exciting action of Dangshen was partially mediated via cholinergic M and N receptors.