Establishment of two-dimensional gel electrophoresis for soybean protein isolate and its application

To optimize the conditions for the establishment of two-dimensional gel electrophoresis(2-DE)of soybean protein isolate(SPI),we investigated Ampholine mixture,anodic and cathodic electrolytes,loading amount of sample,acrylamide concentration,p H gradient and gel staining method in twodimensional gel electrophoresis to optimize the protein imaging conditions in two-dimensional gel.The results of mixed-level design experiments showed that Ampholine,loading amount and gel staining method had significant effect(P<0.05)on 2-DE of SPI.The optimal conditions were Ampholine mixture(pH 3–10+pH 5–7 or pH 4–6+pH 5–7),loading amount of 2 mg sample and silver staining.Although the acrylamide concentration of the gel,the p H gradient,the anodic and cathodic electrolyte solutions had significant statistical effects on the protein separation degree,the complexity of the protein composition and the visibility of the gel images were more inclined to the 12%gel,the 3–10 pH gradient and the H3PO4/NaOH electrolyte.According to the established conditions,the hydrolyzed products of SPI emulsion were determined by 2-DE,and the dynamic changes of protein in the process of enzymatic hydrolysis were described.

Multidimensional autophagy nano-regulator boosts Alzheimer's disease treatment by improving both extra/intraneuronal homeostasis

Intraneuronal dysproteostasis and extraneuronal microenvironmental abnormalities in Alzheimer’s disease(AD)collectively culminate in neuronal deterioration.In the context of AD,autophagy dysfunction,a multi-link obstacle involving autophagy downregulation and lysosome defects in neurons/microglia is highly implicated in intra/extraneuronal pathological processes.Therefore,multidimensional autophagy regulation strategies co-manipulating“autophagy induction”and“lysosome degradation”in dual targets(neuron and microglia)are more reliable for AD treatment.Accordingly,we designed an RP-1 peptide-modified reactive oxygen species(ROS)-responsive micelles(RT-NM)loading rapamycin or gypenoside XVII.Guided by RP-1 peptide,the ligand of receptor for advanced glycation end products(RAGE),RT-NM efficiently targeted neurons and microglia in AD-affected region.This nanocombination therapy activated the whole autophagy-lysosome pathway by autophagy induction(rapamycin)and lysosome improvement(gypenoside XVII),thus enhancing autophagic degradation of neurotoxic aggregates and inflammasomes,and promoting Aβ phagocytosis.Resultantly,it decreased aberrant protein burden,alleviated neuroinflammation,and eventually ameliorated memory defects in 3×Tg-AD transgenic mice.Our research developed a multidimensional autophagy nano-regulator to boost the efficacy of autophagy-centered AD therapy.

Modulating autophagic flux via ROS-responsive targeted micelles to restore neuronal proteostasis in Alzheimer’s disease

Compromised autophagy and defective lysosomal clearance significantly contribute to impaired neuronal proteostasis,which represents a hallmark of Alzheimer’s disease(AD)and other age-related neurodegenerative disorders.Growing evidence has implicated that modulating autophagic flux,instead of inducing autophagosome formation alone,would be more reliable to rescue neuronal proteostasis.Concurrently,selectively enhancing drug concentrations in the leision areas,instead of the whole brain,will maximize therapeutic efficacy while reduing non-selective autophagy induction.Herein,we design a ROS-responsive targeted micelle system(TT-NM/Rapa)to enhance the delivery efficiency of rapamycin to neurons in AD lesions guided by the fusion peptide TPL,and facilitate its intracellular release via ROS-mediated disassembly of micelles,thereby maximizing autophagic flux modulating efficacy of rapamycin in neurons.Consequently,it promotes the efficient clearance of intracellular neurotoxic proteins,β-amyloid and hyperphosphorylated tau proteins,and ameliorates memory defects and neuronal damage in 3×Tg-AD transgenic mice.Our studies demonstrate a promising strategy to restore autophagic flux and improve neuronal proteostasis by rationally-engineered nano-systems for delaying the progression of AD.

Combination of Se-methylselenocysteine,D-α-tocopheryl succinate,β-carotene,and L-lysine can prevent cancer metastases using as an adjuvant therapy

Objective:Primary tumor treatment through surgical resection and adjuvant therapy has been extensively studied,but there is a lack of effective strategies and drugs for the treatment of tumor metastases.Here,we describe a functional product based on a combination of compounds,which can be used as an adjuvant therapy and has well-known mechanisms for inhibiting cancer metastases,improving anti-cancer treatment,and enhancing immunity and antioxidant capacity.Our designed combination,named MVBL,consists of four inexpensive compounds:L-selenium-methylselenocysteine(MSC),D-α-tocopheryl succinic acid(VES),β-carotene(β-Ca),and L-lysine(Lys).Methods:The effects of MVBL on cell viability,cell cycle,cell apoptosis,cell migration,cell invasion,reactive oxygen species(ROS),and paclitaxel(PTX)-combined treatment were studied in vitro.The inhibition of tumor metastasis,antioxidation,and immune enhancement capacity of MVBL were determined in vivo.Results:MVBL exhibited higher toxicity to tumor cells than to normal cells.It did not significantly affect the cell cycle of cancer cells,but increased their apoptosis.Wound healing,adhesion,and transwell assays showed that MVBL significantly inhibited tumor cell migration,adhesion,and invasion.MVBL sensitized MDA-MB-231 breast cancer cells to PTX,indicating that it can be used as an adjuvant to enhance the therapeutic effect of chemotherapy drugs.In mice,experimental data showed that MVBL inhibited tumor metastasis,prolonged their survival time,and enhanced their antioxidant capacity and immune function.Conclusions:This study revealed the roles of MVBL in improving immunity and antioxidation,preventing tumor growth,and inhibiting metastasis in vitro and in vivo.MVBL may be used as an adjuvant drug in cancer therapy for improving the survival and quality of life of cancer patients.