Optimization of preparation process of cationic liposome nanoparticles containing Survivin-siRNA and osthol

Objective:To prepare cationic liposome nanoparticles loaded with survivin-siRNA and Cnidium monnieri based on the ability of liposomes to contain both water-soluble and lipid soluble components.Methods:The preparation technology of Osthol cationic liposomes was optimized by orthogonal test with membrane material ratio,drug lipid ratio,ultrasonic time and steaming temperature as factors.The volume ratio of HA-siRNA to protamine and the ratio of HA-siRNA protamine complex to liposome were investigated by control variable method with potential and particle size as indexes.The particle size and zeta potential were measured by potentiometric particle size analyzer,and the shape was observed by transmission electron microscope;The absorbance of different concentrations of FAM-Survivin-siRNA standard solution was measured by microplate analyzer,and the entrapment efficiency of cationic liposomes loaded with FAM-Survivin-siRNA and osthole was calculated.Results:The optimum preparation conditions of osthole loaded cationic liposomes were as follows:the ratio of membrane to material was 3:1,the ratio of drug to lipid was 1:5,the steaming temperature was 30℃,the ultrasonic time was 70 min,and the encapsulation efficiency was 78.34%.The optimum preparation conditions of osthole loaded cationic liposomes loaded FAM-Survivin-siRNA were as follows:the volume ratio of Survivin-siRNA to protamine was 1:1,Protamine complex 25μg.Add 50μL cationic liposomes.The particle size is 132.3±0.2nm,zeta potential is 43.15±0.05mv,and its shape is irregular round;According to the standard curve,the entrapment efficiency of cationic liposome nanoparticles co loaded with Survivin-siRNA and osthole was 81.34±0.041%.Conclusion:The prepared cationic liposome nanoparticles loaded with Survivin-siRNA and osthole have good encapsulation efficiency,particle size and zeta potential.

Increased Expression of Serum Monocyte Chemoattractant Protein-1,Chemokine(C-C motif)Ligand 3,and Interleukins in Patients With Vitiligo

Objective:Vitiligo is a chronic,acquired depigmentation of the skin,which can be divided into active and stable vitiligo.The stable and active stages are not immutable and often alternate.Identification of internal markers would help to diagnose the active stage of vitiligo,and the purpose of this study was to find potential active markers in the progression of vitiligo.Methods:We compared 30 healthy controls to 60 patients with vitiligo(30 with active disease and 30 with stable disease)regarding the expression of serum monocyte chemoattractant protein(MCP)-1,chemokine(C-C motif)ligand(CCL)3,CCL5,CCL21,interleukin(IL)-6,and IL-17.Specimens were collected from May 2017 to December 2018.Serological detection was carried out by enzyme-linked immunosorbent assay.The difference between groups was compared by Kruskal-Wallis H test and Mann-Whitney U test.Results:We found that the serum levels of MCP-1,CCL3,IL-6,and IL-17 were significantly higher in patients with vitiligo than in healthy controls(P<0.01)and that IL-17 was higher in patients with active vitiligo than in those with stable vitiligo(P<0.01).After 3months of treatment,the serum IL-17 level of patients with active vitiligo was not significantly different from that before treatment(P=0.526);after 6months of treatment,however,the serum IL-17 level had significantly decreased(P<0.01).Conclusion:These findings suggest that the serum IL-17 level can be used as a marker of vitiligo disease activity giving it certain guiding significance for clinical treatment.

Optimization of extraction technology of safflower polysaccharide based on central composite design-response surface methodology

Objective:To explore the best extraction technology of safflower polysaccharide by central composite design response surface methodology and evaluate its quality.Methods:Taking the extraction rate of Carthamus tinctorius polysaccharide as the index,taking the ratio of solid to liquid,extraction time,extraction temperature and extraction times as the investigation factors,based on the single factor experiment,the central composite design-response surface methodology was used to optimize the optimum extraction process of Carthamus tinctorius polysaccharide and verify it.Results:The response surface model was established with the extraction rate of Carthamus tinctorius polysaccharide as dependent variable Y,the ratio of solid to liquid,extraction time and extraction temperature as independent variables X,P<0.0001.The optimum extraction process was as follows:the ratio of solid to liquid was 1:16.69,the extraction temperature was 91.39℃,and the extraction working time was 89.78min.Under these conditions,the extraction rate of safflower polysaccharide can reach 7.45%,The experimental results show that RSD is 1.05%,and the model can well predict the experimental results.Conclusion:Central Composite Design-Response Surface Methodology has the advantages of high extraction rate,simple,effective and reasonable process operation,high stability and high precision,which can be fully applied to the resource management and utilization of safflower polysaccharide.

Allium tuberosum alleviates pulmonary inflammation by inhibiting activation of innate lymphoid cells and modulating intestinal microbiota in asthmatic mice

Objective:This study tests whether long-term intake of Allium tuberosum(AT)can alleviate pulmonary inflammation in ovalbumin(OVA)-induced asthmatic mice and evaluates its effect on the intestinal microbiota and innate lymphoid cells(ILCs).Methods:BALB/c mice were divided into three groups:phosphate buffer saline,OVA and OVA+AT.The asthmatic murine model was established by sensitization and challenge of OVA in the OVA and OVA+AT groups.AT was given to the OVA+AT group by oral gavage from day 0 to day 27.On day 28,mice were sacrificed.Histopathological evaluation of lung tissue was performed using hematoxylin and eosin,and periodic acid-Schiff staining.The levels of IgE in serum,interleukin-5(IL-5)and IL-13 from bronchoalveolar lavage fluid(BALF)were measured by enzyme-linked immunosorbent assay.The ILCs from the lung and gut were detected by flow cytometry.16 S ribosomal DNA sequencing was used to analyze the differences in colon microbiota among treatment groups.Results:We found that long-term intake of AT decreased the number of inflammatory cells from BALF,reduced the levels of IL-5 and IL-13 in BALF,and IgE level in serum,and rescued pulmonary histopathology with less mucus secretion in asthmatic mice.16 S ribosomal DNA sequencing results showed that AT strongly affected the colonic bacteria community structure in asthmatic mice,although it had no significant effect on the abundance and diversity of the microbiota.Ruminococcaceae and Desulfovibrionaceae were identified as two biomarkers of the treatment effect of AT.Moreover,AT decreased the numbers of ILCs in both the lung and gut of asthmatic mice.Conclusion:The results indicate that AT inhibits pulmonary inflammation,possibly by impeding the activation of ILCs and adjusting the homeostasis of gut microbiota in asthmatic mice.