Novel colloidal associations of soyasaponins and lipid components (DPPC,cholesterol) as potential adjuvants for vaccines

Soyasaponins from soybean (Glycine max) represent promising new potent adjuvants for vaccine research because of their immunostimulating properties and weak hemolytic activity. In the present study, saponin microstructures of soyasaponins (soyasaponin Bb, soyasaponin Ab) with lipid components (cholesterol, DPPC (dipalmitoylphosphatidylcholine)) were designed by the lipid film method. In interaction studies between soyasaponins (soyasaponin Ab/Bb) and Langmuir monolayers (model membranes), composed of cholesterol and DPPC, marked interactions between soyasaponins and a pure cholesterol monolayer were observed. No interaction was detected for soyasaponins with a pure DPPC monolayer. The intercalation of soyasaponins in a mixed DPPC/cholesterol (3:1, w/w) monolayer was only observed for the monodesmosidic soyasaponin Bb whereas the second sugar chain of the bidesmosidic soyasaponin Ab impaired the access to the monolayer. Transmission electron microscopy was used for visualizing particle formation of soyasaponins and lipid components. Pseudo-binary systems (soyasaponin Ab/Bb, cholesterol) formed colloidal associations built up from ring-like subunits in the nanometer size range. In pseudo-ternary systems (soyasaponin, cholesterol, DPPC) soyasaponin Bb attacked the liposomal membrane by forming colloidal associations. Colloidal associations in pseudo-ternary systems with soyasaponin Ab, cholesterol and a phospholipid were only observed in the presence of PE (phosphatidylethanolamine) instead of DPPC. In an MTT assay with a HaCaT cell line (keratinocyte cell line) the cell viability was neither affected by the soyasaponins nor by the corresponding formulations. Both the pure soyasaponin solution and the saponin formulations may be promising adjuvant systems for the intradermal vaccine application. Furthermore, interaction studies between the model antigen ovalbumin and colloidal associations of saponins and cholesterol using MST (Microscale Thermophoresis) gave first indications of an antigen binding to colloidal associations. Ex vivo T-cell proliferation in the presence of soyasaponin Ab was confirmed.     

Curcumin ameliorates hepatic chronic inflammation induced by bile duct obstruction in mice through the activation of heme oxygenase-1

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Cutis laxa,exocrine pancreatic insufficiency and altered cellular metabolomics as additional symptoms in a new patient with ATP6AP1-CDG

Congenital disorders of glycosylation (CDG) are genetic defects in the glycoconjugate biosynthesis. > 100 types of CDG are known, most of them cause multi-organ diseases. Here we describe a boy whose leading symptoms comprise cutis laxa, pancreatic insufficiency and hepatosplenomegaly. Whole exome sequencing identified the novel hemizygous mutation c.542 T > G (p.L181R) in the X-linked ATP6AP1, an accessory protein of the mammalian vacuolar H⁺-ATPase, which led to a general N-glycosylation deficiency. Studies of serum N-glycans revealed reduction of complex sialylated and appearance of truncated diantennary structures. Proliferation of the patient's fibroblasts was significantly reduced and doubling time prolonged. Additionally, there were alterations in the fibroblasts' amino acid levels and the acylcarnitine composition. Especially, short-chain species were reduced, whereas several medium- to long-chain acylcarnitines (C14-OH to C18) were elevated. Investigation of the main lipid classes revealed that total cholesterol was significantly enriched in the patient's fibroblasts at the expense of phophatidylcholine and phosphatidylethanolamine. Within the minor lipid species, hexosylceramide was reduced, while its immediate precursor ceramide was increased. Since catalase activity and ACOX3 expression in peroxisomes were reduced, we assume an ATP6AP1-dependent impact on the β-oxidation of fatty acids. These results help to understand the complex clinical characteristics of this new patient.     

Variants of self-assembling peptide,KLD-12 that show both rapid fracture healing and antimicrobial properties

KLD-12 (KLD) is a 12-residue self-assembling peptide that can adopt nano-structures and is known for its tissue-engineering properties. Our objective was to introduce antimicrobial attribute to KLD which would help in preventing secondary infection associated with external application of such tissue engineering materials. Considering the net charge of KLD-12, varying number of cationic arginine residues were added to its N-terminus. KLD variants showed appreciable bactericidal properties without any significant increase in cytotoxicity against tested mammalian cells. Further, these variants adopted β-sheet structures and self-assembled into nano-structures comparable to that of KLD. Interestingly, the KLD variants with two (KLD-2R) and three (KLD-3R) arginine residues added to its N-terminus showed significant osteogenic effect which was comparable or better than the original peptide as evident from the alkaline phosphatase activity assay, mineralized nodule formation and expression of different osteogenic genes. Particularly, application of KLD-2R in rats to the site of a drill-hole (0.8 mm diameter) that was created in the femur metaphysis displayed significantly higher bone regeneration compared to that of KLD. The results demonstrate a simple way to improve biological property of a self-assembling peptide with tissue engineering property.     

参泽舒肝胶囊联合多烯磷脂酰胆碱胶囊治疗非酒精性脂肪肝的临床研究

目的 研究参泽舒肝胶囊联合多烯磷脂酰胆碱胶囊治疗非酒精性脂肪肝的疗效。方法 将南方医科大学珠江医院2017年12月-2019年12月收治的非酒精性脂肪肝患者184例纳入本次研究,并以随机数表法将其分为试验组与对照组,每组92例。予以对照组患者多烯磷脂酰胆碱胶囊治疗,予以试验组患者多烯磷脂酰胆碱胶囊＋参泽舒肝胶囊治疗。比较两组患者治疗后临床疗效、治疗前后肝功能指标及血脂水平、血清学指标水平变化、肝脏B超评分与肝脾CT比值指标、用药不良反应情况等指标。结果 治疗后,试验组患者临床有效率（95.65%）高于对照组（86.96%）（P<0.05）;治疗前,两组患者谷丙转氨酶(aLanine aminotransferase,ALT)、谷草转氨酶(aspartate transaminase,AST)、高密度脂蛋白胆固醇（high density liptein cholesterol,HDL-C）、谷氨酰转肽酶(glutamyl transpeptidase,GGT)、总胆固醇(totalCholesterol,TC)、甘油三酯(triGlyceride,TG)、血清超氧化物歧化酶(superOxide dismutase,SOD)、丙二醛(malondialdehyde ,MDA)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、B超评分与肝脾CT比值等指标比较,差异无统计学意义（P>0.05）;治疗后,两组患者ALT、AST、GGT水平降低,且试验组降低较明显（P<0.05）;两组TC及TG水平降低,HDL-C水平升高（P<0.05）;且试验组改善较明显（P<0.05）;两组患者SOD升高,MDA和TNF-α水平降低（P<0.05）;且试验组改善较明显（P<0.05）;两组患者治疗后B超评分均下降,肝脾CT比值均提升,试验组治疗后B超评分低于对照组,肝脾CT比值高于对照组（P<0.05）。两组患者均无不良反应。结论 参泽舒肝胶囊联合多烯磷脂酰胆碱胶囊治疗非酒精性脂肪肝疗效显著,且安全可行。     

Effects of emulsifier concentration,composition, and order of addition in squalene-phosphatidylcholine oil-in-water emulsions

Development and characterization of stable and biocompatible oil-in-water emulsions is important for improved drug and vaccine delivery. In this work, two-component emulsions consisting of squalene and phosphatidylcholine have been developed. The reproducibility of the manufacturing process is established and production efficiency is improved by altering the order of component addition. The effects of emulsifier concentration and composition on emulsion stability and biocompatibility are assessed through dynamic light scattering, zeta potential measurement, viscosity, and hemolytic activity. High concentrations of egg phosphatidylcholine emulsifier decreased initial particle size and increased initial size polydispersity. However, high emulsifier concentrations also appeared to decrease long-term emulsion stability as well as absolute zeta potential values. Substitution of naturally derived egg phosphatidylcholine with synthetic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) produced an emulsion with similar physicochemical properties and stability.     

Nanomedicine for acute respiratory distress syndrome: The latest application,targeting strategy,and rational design

Acute respiratory distress syndrome (ARDS) is characterized by the severe inflammation and destruction of the lung air–blood barrier, leading to irreversible and substantial respiratory function damage. Patients with coronavirus disease 2019 (COVID-19) have been encountered with a high risk of ARDS, underscoring the urgency for exploiting effective therapy. However, proper medications for ARDS are still lacking due to poor pharmacokinetics, non-specific side effects, inability to surmount pulmonary barrier, and inadequate management of heterogeneity. The increased lung permeability in the pathological environment of ARDS may contribute to nanoparticle-mediated passive targeting delivery. Nanomedicine has demonstrated unique advantages in solving the dilemma of ARDS drug therapy, which can address the shortcomings and limitations of traditional anti-inflammatory or antioxidant drug treatment. Through passive, active, or physicochemical targeting, nanocarriers can interact with lung epithelium/endothelium and inflammatory cells to reverse abnormal changes and restore homeostasis of the pulmonary environment, thereby showing good therapeutic activity and reduced toxicity. This article reviews the latest applications of nanomedicine in pre-clinical ARDS therapy, highlights the strategies for targeted treatment of lung inflammation, presents the innovative drug delivery systems, and provides inspiration for strengthening the therapeutic effect of nanomedicine-based treatment.     

Delayed release phosphatidylcholine as new therapeutic drug for ulcerative colitis – a review of three clinical trials

Importance of the field: As the pathogenesis of ulcerative colitis (UC) is unknown, a causative therapy is lacking. Therefore, some UC patients suffer from disease activity despite symptomatic anti-inflammatory treatment strategies. We claim that reduction of phosphatidylcholine (PC) in colonic mucus impairs the mucosal barrier and, thus, causes attacks of the commensal bacterial flora to induce colitis. Thus, mucus PC substitution could provide a causal therapy for UC.

Areas covered in this review: A delayed released oral PC preparation (rPC) was found to substitute for the lack of PC in rectal mucus. In non-steroid-treated active UC, 53% of rPC-treated patients reached remission compared with 10% of placebo patients (p < 0.001). In a second trial with chronic-active, steroid-dependent UC patients, steroid withdrawal with a concomitant achievement of remission (CAI ≤ 3) or clinical response (≥ 50% CAI improvement) was reached in 15 rPC-treated patients (50%) but only in 3 (10%) placebo patients (p = 0.002).

What the reader will gain: The concept that missing PC in colonic mucus is the main pathogenetic factor for development of UC. PC can be substituted by rPC, which cures the disease in the majority of patients.

Take home message: rPC is, to our knowledge, the first causative therapeutic option for patients with UC.