Preparation, quality evaluation and cell uptake of 1,8-cineole self-microemulsion drug delivery system / 中草药

Objective: To optimize the formulation of 1,8-cineole self-microemulsifying drug delivery system (1,8-Cin-SMEDDS), characterize it and investigate its cell uptake. Methods: By drawing pseudo-ternary phase diagram, the effective self-emulsifying region of 1,8-Cin-SMEDDS was determined, and the preliminary prescription was screened. Taking the particle size and drug loading as the index, the central composite design-response surface method was used to optimize and verify the prescription. Fluorescence microscope was used to observe the uptake of human umbilical vein endothelial cells (HUVEC) injured by high glucose. Results: The results showed that the best prescription of 1,8-Cin-SMEDDS was a mixture of soybean oil (7.5%) and 1,8-Cin (22.5%), HS15 (56%) as emulsifier, ethanol (14%) as co-emulsifier, and dripping pure water to 8 mL to obtain a translucent slightly bluish emulsion. The appearance of spherical droplets was observed by transmission electron microscope, and the average particle size and Zeta potential measured by laser particle size Zeta tester was (131.68 ± 1.44) nm and (-10.03 ± 1.63) mV, respectively; The entrapment efficiency estimated by HPLC was (99.890 ± 0.012)%, and the drug loading was (224.750 ± 0.028) mg/g. The results of HUVEC cell uptake assay showed that the uptake of 1,8-Cin-SMEDDS by cells was higher than that of free 1,8-Cin. Conclusion: The preparation method of 1,8-Cin-SMEDDS is simple and reproducible. The obtained method has good appearance, high entrapment efficiency, stable physical, and chemical properties, which can also promote cell uptake.

Indication of bloodletting therapy based on multi-dimensional evidence assessment / 中国针灸

The indication of bloodletting therapy was determined based on the multi-dimensional evidence assessment, which could provide guidance for the clinical application of bloodletting therapy. The literature of bloodletting therapy was comprehensively collected by retrieval in CNKI, Wanfang and VIP databases (until February 23, 2019), modern books in Library of Tianjing University of TCM and the (Fifth Edition). The disease spectrum of bloodletting therapy was determined by self-designed questionnaire survey e-mailed to relevant experts. The indication of bloodletting therapy was determined by Delphi expert meeting. As a result, 746 pieces of ancient literature and 32 775 modern literature were included. The indications of bloodletting therapy based on the multi-dimensional evidence assessment include herpes zoster, acne, acute tonsillitis, vascular headache, varicose veins of lower extremities, acute lumbar sprain, early erysipelas, wheat swelling, exogenous fever of children, stroke, which are mainly the syndromes of blood stasis, toxin, excess and heat.

Preparation of Oxymatrine Phospholipid Complex Solid Lipid Nanoparticles Lyophilized Powder and Evaluation of Its Quality / 中国实验方剂学杂志

Objective: To prepare oxymatrine phospholipid complex solid lipid nanoparticles(OMT-PC-SLN) lyophilized powder and evaluate its pharmaceutical properties. Method: Pseudo-ternary phase diagram was employed to optimize the formula of microemulsion;single factor experiments were adopted to optimize the formulation process of OMT-PC-SLN lyophilized powder with encapsulation efficiency as index;the morphology of this preparation was observed by transmission electron microscope(TEM).The particle size was measured by particle size analyzer and the in vitro release performance of OMT-PC-SLN lyophilized powder was examined. Result: Optimal formulation process was as following:taking soybean phospholipid and polyethylene glycol 15-hydroxystearate(Kolliphor HS 15) as the emulsifier,ethanol as co-emulsifier,ratio of emulsifier to co-emulsifier(Km)=3:2,oil phase:(emulsifier+co-emulsifier)=1:9,oxymatrine phospholipid complex-stearic acid-soybean phospholipid-Kolliphor HS 15-ethanol(30:100:180:360:360);taking 50 mL of 4%mannitol solution as the external aqueous phase,ice bath stirring at 1 000 r·min-1 and solidifying for 1 h,precooled at -20℃ for 24 h,took out and dried for 24 h.OMT-PC-SLN lyophilized powder was spherical in appearance with encapsulation efficiency of (38.09±1.24)%,average particle size of 785.5 nm,polydispersity coefficient(PDI) of 0.456 and the Zeta potential of -24.82 mV.The cumulative release rates of OMT-PC-SLN lyophilized powder were 72.63%at 2 h and 98.42%at 12 h;the cumulative release rate of oxymatrine(crude drug) was 98.60%at 2 h. Conclusion: This optimized formulation process of OMT-PC-SLN lyophilized powder is stable with good repeatability;compared with oxymatrine,OMT-PC-SLN lyophilized powder has a certain sustained-release effect.

Role and mechanism of the expression of matrix metalloproteinase 9 and tumor necrosis factor α in upper and lower respiratory tract inflammation in rats / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the role and mechanism of matrix metalloproteinase 9 (MMP-9) and tumor necrosis factor α (TNF-α) in the pathogenesis of allergic rhinitis (AR) and asthma(AS).</p><p><b>METHODS</b>The rat models of AR and AS were made by injecting ovalbumin. The infiltration of eosinophils and mast cells were detected by hematoxylin-eosin (HE) staining and toluidine blue staining respectively, and the expression of MMP-9 and TNF-α in nasal mucosa and lung tissue were examined by immunohistochemical staining (SP method). The relationship of their expression with upper and lower respiratory tract inflammation was analyzed. SPSS 13.0 software was used to analyze the data.</p><p><b>RESULTS</b>The numbers of MMP-9 positive inflammatory cells in nasal mucosa and lung tissue of AR were (154.8 ± 12.0) and (124.0 ± 8.2), (43.2 ± 7.6) and (34.5 ± 5.0) in the control groups, the difference was significant (t value were 24.260, 29.525 respectively, all P < 0.05). The numbers of MMP-9 positive inflammatory cells in nasal mucosa and lung tissue of AS were (149.9 ± 11.7) and (120.1 ± 7.3), (48.6 ± 7.6) and (39.1 ± 5.2) in control groups, the difference was significant (t value were 22.929 and 28.530 respectively, all P < 0.05). The numbers of TNF-α positive inflammatory cells in nasal mucosa and lung tissue of AR were (188.8 ± 17.0), and (134.8 ± 7.9), (57.6 ± 23.3) and (40.3 ± 8.2) in control groups, the difference was significant (t value were 13.836 and 26.220, all P < 0.05). The numbers of TNF-α positive inflammatory cells in nasal mucosa and lung tissue of AS were (179.2 ± 15.4) and (153.5 ± 10.1), (70.5 ± 33.1) and (33.8 ± 14.0) in control groups, the difference was significant (t value were 9.412 and 21.858, all P < 0.05). There was a correlation between the expression of MMP-9 and TNF-α in nasal mucosa and lung tissue of AR (r values were 0.893 and 0.700 respectively, P values were 0.001 and 0.024, respectively) and AS (r values were 0.692 and 0.644 respectively, P values were 0.027 and 0.044 respectively) groups.</p><p><b>CONCLUSIONS</b>The inflammation is similar between AR and AS. The MMP-9 and TNF-α may play an important role in the pathogenesis of upper and lower respiratory tract inflammation.</p>

A preliminary study of radionuclide imaging of the nasal lymphatic system in nasal polyps / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the role of nasal mucosal lymphatic drainage in the pathogenesis of nasal polyps.</p><p><b>METHODS</b>There were 25 cases in the experimental group who had nasal polyps (which was further divided into Malm-1, Malm-2, Malm-3 level 3 subgroups) and 6 cases in the control group, including thyroid cancer and laryngeal cancer patients who had normal nasal structure. The nasal polyps in the experimental group and the middle turbinate in the control group were injected with a radionuclide and a radionuclide imaging technique was used to image the nasal mucosal lymphatics. The lymphatic drainage status of the nasal mucosa through the imaging results was analysed.</p><p><b>RESULTS</b>The T/NT ratio (radioactivity counting) of the region of interest (ROI) was 20. 66 +/- 1.89 in the control group and 29. 33 +/- 6.34 in the experimental group. The difference was significant (t = 3.275, P < 0.05). The T/NT ratio of the ROI was 24.40 +/- 3.19 in the Malm-1 level group, 29.31 +/- 3.39 in the Malm-2 level group, 39.21 +/- 3.15 in the Malm-3 level group. The differences of qualitative analysis were significant (F = 38. 980, P < 0.05). The quantitative analysis showed that at the injection site, signs of lymphatic development and drainage were not found in the control group or experimental group, but the phenomenon of contrast media retention existed at the injection site in the experimental group.</p><p><b>CONCLUSION</b>Lymphatic drainage dysfunction exists in patients with nasal polyps, and it may play a role in the pathogenesis of nasal polyps.</p>

Protective effect of nitric oxide against hydrogen peroxide-induced hearing loss / 生理学报

Previous research showed that reactive oxygen species (ROS) play an important role in ototoxity. The present research was to investigate whether nitric oxide, an important neurotransmitter in the inner ear, could prevent hydrogen peroxide-induced hearing loss through the nitric oxide/cyclic GMP pathway in guinea pig cochlea. Fifty adult pigmented guinea pigs (250~350 g) of either sex with positive prier reflex were randomly divided into five groups. All of the animals underwent whole cochlear perfusion for two hours. The solution that was perfused into the cochlear of different group was artificial perilymph (AP) for group 1200 micromol/L H2O2 for group 2100 micromol/L L-Arg for group 3, H2O2+L-Arg for group 4 and H2O2+L-Arg+L-NNA for group 5 respectively. Compound action potential (CAP, evoked by click) and cochlear microphonic (CM, evoked by tone burst) were recorded every thirty minutes to show the effects of different reagents on cochlear function. In order to assess cell viability after perfusion, the fluorescent dyes Hoechst that stains all cell nuclei and propidium iodide (PI) that specifically stains nuclei of dead cells, were used. The CAP threshold shifts and CM amplitude decreased after perfusion with H2O2+L-Arg. They were significantly lower than those of H2O2 group. No obvious cell death was noticed after H2O2+L-Arg perfusion, while only 54% of hair cells were alive after H2O2 perfusion. There were no significant differences between the group of H2O2 and that of H2O2+L-Arg+L-NNA group. Our results suggest that nitric oxide may partly be able to protect guinea pigs from hydrogen peroxide-induced hearing loss.