Synergism of vesicle trafficking and cytoskeleton during regulation of plant growth and development: A mechanistic outlook.

The cytoskeleton is a fundamental component found in all eukaryotic organisms, serving as a critical factor in various essential cyto-biological mechanisms, particularly in the locomotion and morphological transformations of plant cells. The cytoskeleton is comprised of three main components: microtubules (MT), microfilaments (MF), and intermediate filaments (IF). The cytoskeleton plays a crucial role in the process of cell wall formation and remodeling throughout the growth and development of cells. It is a highly organized and regulated network composed of filamentous components. In the basic processes of intracellular transport, such as mitosis, cytokinesis, and cell polarity, the plant cytoskeleton plays a crucial role according to recent studies. The major flaws in the organization of the cytoskeletal framework are at the root of the aberrant organogenesis currently observed in plant mutants. The regulation of protein compartmentalization and abundance within cells is predominantly governed by the process of vesicle/membrane transport, which plays a crucial role in several signaling cascades.The regulation of membrane transport in eukaryotic cells is governed by a diverse array of proteins. Recent developments in genomics have provided new tools to study the evolutionary relationships between membrane proteins in different plant species. It is known that members of the GTPases, COP, SNAREs, Rabs, tethering factors, and PIN families play essential roles in vesicle transport between plant, animal, and microbial species. This Review presents the latest research on the plant cytoskeleton, focusing on recent developments related to the cytoskeleton and summarizing the role of various proteins in vesicle transport. In addition, the report predicts future research direction of plant cytoskeleton and vesicle trafficking, potential research priorities, and provides researchers with specific pointers to further investigate the significant link between cytoskeleton and vesicle trafficking.

Purification and extraction of fibroblast growth factor 21 (FGF-21) protein by sumo fusion in Escherichia coli.

Fibroblast growth factor 21 has recently discovered its pivotal role in glucose, lipid metabolism and regulation of energy homeostasis. Further, it has helped in forming great strides for treatment of chronic diseases like diabetes and inflammation. FGF-21 was sub-cloned into the SUMO vector and was induced for expression in Escherichia coli Rosetta. The recombinant plasmid was transformed into Escherichia coli strain. FGF-21 was induced by IPTG and purified by Ni-NTA agarose (Nickel-nitrilotriacetic acid) column. The purified fusion protein was cleavaged by SUMO protease I to obtain recombinant FGF-21 with high purity. The purified protein was tested for its biological activity of FGF-21. HepG2 cell model was used to detect the regulation of glucose uptake activity of FGF-21 and were further treated with different concentrations of FGF-21.The residual glucose content in medium was measured using the glucose oxidase-peroxidase method. The results indicated that FGF-21 protein had a role in regulating the glucose uptake on HepG2 cells and the effect was significantly dose-dependent manner. In order to further verify whether purified FGF-21 protein obtained has biological activity in diabetic model. Studies have demonstrated that FGF-21 had a greater efficacy in dropping blood glucose in streptozotocin induced diabetic mice.

Therapeutic effect and mechanism of combined use of FGF21 and insulin on diabetic nephropathy.

Although FGF21 ameliorates diabetic nephropathy (DN), the efficacy is not satisfactory. Studies demonstrate that FGF21 combined with Insulin exhibits reciprocal sensitization on glucose and lipid metabolism in mice with type 2 diabetes. However, therapeutic effect of combined use of FGF21 and Insulin on DN has not been reported. Therefore, this study explored therapeutic effect and mechanism of combined use of FGF21 and Insulin on DN. Our results showed that compared with Insulin or FGF21 alone, FGF21 combined with Insulin further ameliorated blood glucose, HbAlc, OGTT, renal function, liver function, blood lipid, histopathological changes, oxidative stress and AGEs in the mice of DN (BKS-Leprem2Cd479/Gpt). Moreover, FGF21 combined with Insulin further reduced expressions of IL-1ß, IL-6, TNF-α via promoting M1 type macrophage into M2 type macrophage. Results of real-time PCR and Western blot showed that FGF21 combined with Insulin upregulated the expressions of autophagy related genes LC3-Ⅱ and BCL-1. Mesangial cells play an important role in the pathological changes of DN mice. However, the effect of FGF21 on mesangial cells has not been reported. In this study, d-glucose was used in high glucose (HG) model in mesangial cells. The results showed that FGF21 significantly reduced the levels of OS, AGEs and cell overproliferation. Meanwhile, FGF21 significantly ameliorated autophagy level via upregulating the phosphorylation of AMPK and downregulating phosphorylation of mTOR. These effects were reversed in siRNA-ß-klotho transfected mesangial cells. In conclusion, our results demonstrate that combination FGF21 with Insulin exhibits a better therapeutic effect on DN compared with FGF21 or Insulin alone. This study provides a theoretical basis for combined used of FGF21 and Insulin as a new treatment for DN and further provides theoretical support for application of FGF21 in treatment of DN.

FGF21 ameliorates hepatic fibrosis by multiple mechanisms.

BACKGROUND: Previous study reports that fibroblast growth factor 21 (FGF21) could ameliorate hepatic fibrosis, but its mechanisms have not been fully investigated. METHODS AND RESULTS: In this study, three models were used to investigate the mechanism by which FGF21 alleviates liver fibrosis. Hepatic fibrosis animal models were respectively induced by CCL4 and dimethylnitrosamine. Our results demonstrated that liver index and liver function were deteriorated in both models. Hematoxylin and eosin and Masson's staining showed that the damaged tissue architectonics were observed in the mice of both models. Treatment with FGF21 significantly ameliorated these changes. ELISA analysis showed that the serum levels of IL-1ß, IL-6 and TNF-α were significantly elevated in both models. However, administration of FGF21 significantly reduced these inflammatory cytokines. Real-time PCR and Western blot analysis showed that treatment with FGF21 significantly decreased mRNA and protein expressions of collagenI, α-SMA and TGF-ß. Platelet-derived growth factor-BB (PDGF-BB) stimulant was used to establish the experimental cell model in hepatic stellate cells (HSCs). Real-time PCR and Western blot analysis demonstrated that the expression of collagenI and α-SMA were significantly upregulated by this stimulant in model group. Interestingly, our results showed that mRNA and protein expressions of leptin were also significantly induced in PDGF-BB treated HSCs. Administration of FGF21 significantly reduced leptin expression in a dose dependent manner and these effects were reversed in siRNA (against ß-klotho) transfected HSCs. Furthermore, the leptin signaling pathways related protein p-ERK/t-ERK, p-STAT3/STAT3 and TGF-ß were significantly downregulated by FGF21 treatment in a dose dependent manner. The expressions of SOCS3 and Nrf-2 were enhanced by treatment with FGF21. The underlying mechanism may be that FGF21 regulates leptin-STAT3 axis via Nrf-2 and SOCS3 pathway in activated HSCs. CONCLUSIONS: FGF21 ameliorates hepatic fibrosis by multiple mechanisms.

Screening Potential Probiotic Characteristics of Lactobacillus brevis Strains In Vitro and Intervention Effect on Type I Diabetes In Vivo.

Diabetes has become the third most serious threat to human health, after cancer and cardiovascular disease. Notably, Lactobacillus brevis is the most common species of LAB that produces γ-aminobutyric acid (GABA). The aim of this study is to clarify the effect of time, strain types, antibiotic concentrations, different levels of pH, and intestinal juices in aerobic or anaerobic conditions and the effect of interactions between these factors on the potential properties of KLDS 1.0727 and KLDS 1.0373, furthermore, antagonistic activity against foodborne pathogens. Moreover, another aim is to study the capability of KLDS 1.0727 and KLDS 1.0373 strains as gad gene carriers to express GABA that reduce the risk of type 1 diabetes in C57BL/6 mice as diabetic models. The obtained results exhibited the surprising tolerance of Lactobacillus brevis strains in vitro digestion models mimicking the conditions of the gastrointestinal tract, further, large antagonistic activity against foodborne pathogeneses. In vivo results displayed the significant effect on glucose level reduction, blood plasma, and histological assays of mice organs. As recommended, the use of Lactobacillus brevis strains should be widely shared in the market as a natural source of GABA in pharmaceutical and food applications.

A novel fusion protein attenuates collagen-induced arthritis by targeting interleukin 17A and tumor necrosis factor α.

This study was undertaken to generate a novel dual targeting fusion protein (DTF), targeting tumor necrosis factor α (TNF-α) and interleukin 17A (IL-17A), and determine its anti-arthritis properties in vitro and in vivo. DTF consisted of an anti-IL-17A single chain variable fragment, a soluble TNF receptor 1, and an Fc fragment. Both clinical and histopathological evaluations suggest that DTF and etanercept can ameliorate collagen induced arthritis. However, the arthritis severity score of DTF-treated mice was lower than that of etanercept-treated mice. In addition, DTF was more potent than etanercept in decreasing the ratio of RANKL/OPG in the serum and rebalancing the population ratio of Treg/Th17 cells in the spleens. In vitro, IL-17A and TNF-α had synergistic effects in inducing the expression of inflammatory cytokines in fibroblast-like synoviocyte from RA patients (RA-FLS), human leukemia (THP-1), and rheumatoid synovial fibroblast (MH7A). IL-17A and TNF-α also had synergistic effects in inducing proliferation and migration of MH7A cells. However, we observed that DTF was more efficient than etanercept in suppressing these synergistic effects. Our results demonstrate that DTF is highly efficient in the treatment of arthritis and has the potential to overcome the limited therapeutic responses obtained with single cytokine neutralization.

FGF-21 Elevated IL-10 Production to Correct LPS-Induced Inflammation.

Fibroblast growth factor 21 (FGF-21) has been previously judged as a major metabolic regulator. In this paper, we show that FGF-21 has a potential role in anti-inflammation and immunoregulation. In vivo, treatment with exogenous FGF-21 can alleviate LPS-induced inflammation. In vitro, FGF-21 inhibited LPS-induced IL-1ß expression in THP-1 cells. Furthermore, besides the NF-κB pathway, the mechanism of action of FGF-21 was observed to involve the elevation of IL-10 in the ERK1/2 pathway. This study clearly indicates that FGF21 can be used as an attractive target for the management of inflammatory disorders. This piece of research indicates that FGF-21 could have much value in the management of inflammatory disorders.

Efficacy of a combination of high and low dosage of PEGylated FGF-21 in treatment of diabetes in db/db mice.

The aim of this study is to explore a new method-high dose starting and low dose maintaining for PEGylated Fibroblast growth factor 21 (pFGF-21) treatment. Db/db mice were initially treated with pFGF-21 of high dose, then treated with pFGF-21 of low doses. The mice were treated with pFGF-21 at initial dose of 1.0mg/kg for 14days, then treated with pFGF-21 at maintenance doses of 0.125/0.250/0.375/0.500mg/kg for 30days. The hypoglycemic and hypolipidemic effects of pFGF-21 of different maintenance doses were compared. The pharmacological efficacy of the maintenance doses of pFGF-21 was evaluated by blood glucose levels, oral glucose tolerance test, glycosylated hemoglobin levels, insulin levels, body weight, lipid profile parameters, the mRNA expressions of glycolysis-related genes, the mRNA expressions of gluconeogenesis-related genes and the mRNA expressions of lipid metabolism-related genes. Results showed that in comparison to the mice treated only with initial dose, the treatment with pFGF-21 at maintenance doses of 0.125/0.250/0.375/0.500mg/kg exhibited favorable efficacy in lowering blood glucose levels and glycosylated hemoglobin levels, thus alleviating insulin resistance and improving dyslipidemia. However, among all of the maintenance doses, the dose of 0.125mg/kg was less effective than the other maintenance doses. These results suggest that using the treatment method of high dose of PEGylated FGF-21 in the start and low dose maintaining results in favorable control of the glycolipid metabolic balance. This study provides a new method for PEGylated FGF-21 treatment which is beneficial to promote the clinical application of PEGylated FGF-21.

Selenium Deficiency Activates Heat Shock Protein Expression in Chicken Spleen and Thymus.

Heat shock proteins (Hsps) are protective proteins present in nearly all species; they are used as biomarkers of various stress conditions in humans, animals, and birds. Selenium (Se) deficiency, which can depress the production of Hsps, can cause chicken tissue injuries. To investigate Hsp production, mRNA, and protein levels in Se-deficient chicken spleens and thymuses, a total of 180 1-day-old sea blue white laying hens (90 chickens/group) were harvested in two groups (the control group and the Se-deficient group) in 15, 25, 35, 45, and 55 days, respectively. The results showed that mRNA levels of Hsp27, Hsp40, Hsp60, Hsp70, and Hsp90 were significantly increased in the spleens and thymuses of the Se-deficient group compared to the control group. Further protein levels of Hsp60, Hsp70, and Hsp90 were also significantly increased in the spleen and thymus of the Se-deficient group compared to the control group. Meanwhile, the spleen expression ratio of Hsp40 mRNA level and Hsp70 protein level were higher in the Se-deficient group than other proteins. In the thymus, the Hsp90 mRNA level and Hsp60 protein expression level were the highest level in the Se-deficient group among other proteins. Based on these results, we concluded that Se deficiency could induce a protective stress response in chicken by means of promoting the mRNA and protein expression of Hsps, thus easing the effects of Se deficiency to some extent.