Systemic transplantation of human umbilical cord derived mesenchymal stem cells-educated T regulatory cells improved the impaired cognition in AβPPswe/PS1dE9 transgenic mice.

Alzheimer's disease (AD) is one of most prevalent dementias, which is characterized by the deposition of extracellular amyloid-beta protein (Aβ) and the formation of neurofibrillary tangles within neurons. Although stereotaxic transplantation of mesenchymal stem cells (MSCs) into the hippocampus of AD animal model as immunomodulatory cells has been suggested as a potential therapeutic approach to prevent the progress of AD, it is invasive and difficult for clinical perform. Systemic and central nervous system inflammation play an important role in pathogenesis of AD. T regulatory cells (Tregs) play a crucial role in maintaining systemic immune homeostasis, indicating that transplantation of Tregs could prevent the progress of the inflammation. In this study, we aimed to evaluate whether systemic transplantation of purified autologous Tregs from spleens of AβPPswe/PS1dE9 double-transgenic mice after MSCs from human umbilical cords (UC-MSCs) education in vitro for 3 days could improve the neuropathology and cognition deficits in AβPPswe/PS1dE9 double-transgenic mice. We observed that systemic transplantation of autologous Tregs significantly ameliorate the impaired cognition and reduced the Aβ plaque deposition and the levels of soluble Aβ, accompanied with significantly decreased levels of activated microglia and systemic inflammatory factors. In conclusion, systemic transplantation of autologous Tregs may be an effective and safe intervention to prevent the progress of AD

Assessment of Binding Interaction between Bovine Lactoferrin and Tetracycline Hydrochloride: Multi-Spectroscopic Analyses and Molecular Modeling

In this paper, the interaction between bovine lactoferrin (bLf) and tetracycline hydrochloride (TCH) was researched by microscale thermophoresis (MST), multi-spectroscopic methods, and molecular docking techniques. Normal fluorescence results showed that TCH effectively quenched the intrinsic fluorescence of bLf via static quenching. Moreover, MST confirmed that the combination force between bLf and TCH was very strong. Thermodynamic parameters and molecular docking further revealed that electrostatic forces, van der Waals, and hydrogen bonding forces played vital roles in the interaction between bLf and TCH. The binding distance and energy transfer efficiency between TCH and bLf were 2.81 nm and 0.053, respectively. Moreover, the results of circular dichroism spectra (CD), ultraviolet visible (UV-vis) absorption spectra, fluorescence Excitation-Emission Matrix (EEM) spectra, and molecular docking verified bLf indeed combined with TCH, and caused the changes of conformation of bLf. The influence of TCH on the functional changes of the protein was studied through the analysis of the change of the bLf surface hydrophobicity and research of the binding forces between bLf and iron ion. These results indicated that change in the structure and function of bLf were due to the interaction between bLf and TCH

Transplantation of UC-MSCs educated CD4⁺CD25⁺ T regulatory cells regulated the levels of cytokines in the plasma of Tg mice.

<div><p><b>A & B</b> Transplantation of UC-MSCs educated CD4⁺CD25⁺ T regulatory cells significantly improved the plasma level of TGFβ1 (A) and IL-10 (B).</p> <p><b>C</b> Transplantation of UC-MSCs educated CD4⁺CD25⁺ T regulatory cells significantly reduced the plasma level of interferon-γ. Data represented mean (±SD) of four experiments. **p<0.01.</p></div

Transplantation of UC-MSCs educated CD4⁺CD25⁺ T regulatory cells inhibited microglia activation and reduced the the level of Aβ in Tg mice.

<div><p>Immunofluorescence using a primary antibody (Iba-1) in conjunction with TRITC-conjugated secondary antibody was used to label microglia. Thiofalvin S staining was used to label the Aβ plaque.</p> <p><b>A & B</b> Representative result of activated microglia in the brain of AβPPswe/PS1dE9 transgenic mice with systemic transplantation of UC-MSCs educated CD4⁺CD25⁺ T regulatory cells (A) and PBS (B).</p> <p><b>C</b> The bar showed that systemic transplantation of UC-MSCs educated CD4⁺CD25⁺ T regulatory cells significantly reduced the number of the Iba-1 positive cells in the brain of Tg mice. Data from 10 serial sections at an interval of every 5^th section through the bilateral cortex and hippocampus were summed to derive representative values for each animal for positive cells and 6 mice per group. Data are reported as mean ±S.E.M. *p<0.05.</p> <p><b>D & E</b> Representative results of Aβ plaque in the cortex of βPPswe/PS1dE9 transgenic mice with systemic transplantation of UC-MSCs educated CD4⁺CD25⁺ T regulatory cells (D) and PBS (E).</p> <p><b>G&H</b> Representative results of Aβ plaque in the hippocampus of AβPPswe/PS1dE9 transgenic mice with systemic transplantation of UC-MSCs educated CD4⁺CD25⁺ T regulatory cells (G) and PBS (H).</p> <p><b>F&I</b> The bar showed that Systemic transplantation of UC-MSCs educated CD4⁺CD25⁺ T regulatory cells significantly reduced the area of Aβ plaque in the cortex (F) and hippocampus (I). Data from 10 serial sections at an interval of every 5^th section through the bilateral cortex and hippocampus were summed to derive representative values for each animal for total plaque area and 6 mice per group. Data are reported as mean ±S.E.M. **p<0.01.</p> <p><b>J&K</b> The bar showed that systemic transplantation of UC-MSCs educated CD4⁺CD25⁺ T regulatory cells significantly reduced the level of the whole brain soluble Aβ1-42 (J) and Aβ1-40 (K) by ELISA test. Data from 6 mice are reported as mean ±S.E.M. *p<0.05.</p></div

Protective Effects of Polyphenol Ex-tracts from Sea Buckthorn (Hippophaė rhamnoides L.) on Rat Hearts

Abstract The objective of present study was to investigate the antioxidative and cardioprotective effects of polyphenol extracts from sea buckthorn (PESB) against myocardial ischemia reperfusion injury (MIRI). PESB was analyzed using HPLC-DAD and their antioxidant activities were evaluated by measuring the intrinsic free radicals cavenging capacity and inhibiting the formation of hydroxyl radical capacity. Experiments on the isolated rat hearts were carried out according to the improved Langendorff retrograde perfusion technology. Coronary effluent of lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CK-MB) activities were analyzed to determine the extent of myocardial injury. Protein expression levels of Beclin-1, microtubule-associated protein 1 light chain 3 (LC3) I and II were measured to evaluate the degree of autophagy. In conclusion, coupled results from both in vivo and in vitro experiments have confirmed that PESB has potential health implications in both prevention and amelioration of MIRI, which may be attributed to the inhibition of autophagy

Methyllycaconitine alleviates amyloid-β peptides-induced cytotoxicity in SH-SY5Y cells.

Alzheimer's disease (AD) is a chronic progressive neurodegenerative disorder. As the most common form of dementia, it affects more than 35 million people worldwide and is increasing. Excessive extracellular deposition of amyloid-β peptide (Aβ) is a pathologic feature of AD. Accumulating evidence indicates that macroautophagy is involved in the pathogenesis of AD, but its exact role is still unclear. Although major findings on the molecular mechanisms have been reported, there are still no effective treatments to prevent, halt, or reverse Alzheimer's disease. In this study, we investigated whether Aβ25-35 could trigger an autophagy process and inhibit the growth of SH-SY5Y cells. Furthermore, we examined the effect of methyllycaconitine (MLA) on the cytotoxity of Aβ25-35. MLA had a protective effect against cytotoxity of Aβ, which may be related to its inhibition of Aβ-induced autophagy and the involvement of the mammalian target of rapamycin pathway. Moreover, MLA had a good safety profile. MLA treatment may be a promising therapeutic tool for AD

Aβ25–35-induced growth inhibition of SH-SY5Y cells is mediated by autophagy.

<p>(A) Annexin V/PI staining of apoptosis of SH-SY5Y cells treated with different doses of Aβ_25–35 for 24 h. (B) Hoechst staining of apoptosis of SH-SY5Y cells treated with different doses of Aβ_25–35 for 24 h. (C) Immunoblotting of Beclin 1 and LC3 expression with SH-SY5Y cell lysates. Cells were treated with 10 µM Aβ_25–35 for additional 24 h after transfection with random siRNA or beclin 1 siRNA for 48 h. (D) MTT assay of cell viability of SH-SY5Y cells treated with 10 µM Aβ_25–35 for 24 h after transfection with random siRNA or beclin 1 siRNA for 48 h.</p

Aβ25–35 induces autophagy in SH-SY5Y cells.

<p>(A) Western blot analysis of LC3 protein expression in SH-SY5Y cells treated with different doses of Aβ_25–35 and quantification, β-actin was a loading control. (B) Immunofluorescence microscopy of punctate pattern of LC3 localization in SH-SY5Y cells treated with Aβ_25–35. (C) Electron micrographs of SH-SY5Y cells treated with Aβ_25–35 for 24 h. (D) Fluorescence microscopy of the formation of acidic vesicles after MDC staining in SH-SY5Y cells treated with Aβ_25–35 for 4 h. (E)Western blot analyses of the protein expression of LC3 and p62 in cells treated with Aβ_25–35 with and without chloroquine.</p