Effect of nano-selenium loaded with lycium barbarum polysaccharide on the proliferation of lens epithelial cells after UVB damage in vitro

AIM: To investigate the effect of nano-selenium loaded with different concentrations of lycium barbarum polysaccharide (LBP-SeNPs) on the proliferation of human lens epithelial cells (HLECs) from UV irradiation. METHODS: LBP-SeNPs were prepared and their particle size was detected. HLECs (SRA01/04) were irradiated with UVB for different time (0, 10, 20, 30, 40, 50, 60min) to construct a damaged model, the survival rate of cells was determined by methylthiazol tetrazolium (MTT) assay. The 4',6-Diamidine-2'-phenylindole dihydrochloride (DAPI) staining was used to observe the status of cell nucleus and drug entering cytoplasm through cell membrane in SRA01/04 cells after adding LBP-SENPS loaded with coumarin fluorescence agent 24h under fluorescence microscope. SRA01/04 normal and UVB-damaged cells were treated with different amounts of LBP-SeNPs at different concentrations, cells proliferation were observed. RESULTS: The particle size of LBP-SeNPs was stable in the range of 150-200 nm. The survival rate changes with time after UVB irradiation were statistically significant. The 10min of UVB exposure as the time was chosen to construct the cell damage model. With DAPI staining, LBP-SeNPs were observed to enter the cytoplasm through the cell membrane under fluorescence inverted microscope. Cytotoxicity of SRA01/04 at different concentrations of LBP-SeNPs were measured. Cell survival rate was statistically different compared with the control group. The higher the loading concentration of LBP in nano-Se drugs was, the higher the cell proliferation rate was (P<0.05). The lower the concentration of LBP-SeNPs, the higher the cell proliferation rate, showing a negative growth trend (P<0.05). The group with the highest average cell proliferation rate was 0.5 µmol/L 2.0 mg/mL LBP-SeNPs (128.80%). When the 2.0 mg/mL LBP-SeNPs group was selected for cell photography, the cell density was higher at 0.5 μmol/L. With the increase of concentration, SRA01/04 cells appeared more cytoplasm dehydration, cell shrinkage and apoptotic bodies, and cell density decreased. CONCLUSION: LBP-SeNPs has moderate particle size and good stability. LBP-SeNPs can protect HLECs (SRA01/04) from UVB-induced damage, and the cell proliferation rate is further increased with increasing the amount of loaded LBP and decreasing nano-selenium concentration.

Terminalia chebula reduces melanoma cell proliferation and increases apoptosis by inhibiting tyrosinase

Recently, research on using plant-derived drugs as anticancer agents has been greatly developed. Terminalia chebula (T. chebula) Retz., the mature fruit of T. chebula, contains active ingredients that have antioxidant, antimicrobial, and antitumor effects. Because traditional Chinese medicine components have unstable physical and chemical properties, such as poor solubility, fast decomposition, and high irritation, drug nanocarriers that effectively increase their solubility and reduce irritation are desirable. Additionally, T. chebula fruit can inhibit tyrosinase activity. This study aimed to determine whether T. chebula nanostructured lipid carriers can regulate the proliferation and apoptosis of melanoma cells by inhibiting tyrosinase activity. Tyrosinase activity in drug-treated melanoma cells was detected by MTT and DOPA oxidation assays, and cell proliferation rate was observed. Western blot (MTT) was used to detect the activity of apoptotic factors, and flow cytometry was used to determine the apoptotic rate. The results showed that tyrosinase activity and cell proliferation rate decreased with increasing drug concentration in cultured cells. Furthermore, the inhibitory effect was concentration-dependent. Additionally, cells cultured with a high concentration of T. chebula nanostructured lipid carriers could activate apoptotic factors and increase cell apoptosis. Thus, this study showed that T. chebula fruit nanostructured lipid carriers could inhibit the proliferation of melanoma cells and increase apoptosis by inhibiting tyrosinase activity, demonstrating a new plant-derived drug for melanoma treatment.

Over-Expression of the Cell-Cycle Gene LaCDKB1;2 Promotes Cell Proliferation and the Formation of Normal Cotyledonary Embryos during Larix kaempferi Somatic Embryogenesis

Somatic embryogenesis is an effective tool for the production of forest tree seedlings with desirable characteristics; however, the low initiation frequency and productivity of high-quality mature somatic embryos are still limiting factors for Larix kaempferi (Japanese larch). Here, we analyzed the expression pattern of L. kaempferi cyclin-dependent kinase B 1;2 (LaCDKB1;2) during somatic embryogenesis in L. kaempferi and its relationship with the cell proliferation rate. We also analyzed the effect of LaCDKB1;2 over-expression on somatic embryo quality. The results revealed a positive correlation between LaCDKB1;2 expression and the cell proliferation rate during the proliferation stage. After LaCDKB1;2 over-expression, the proliferation rate of cultures increased, and the number of somatic embryos in transgenic cultures was 2.69 times that in non-transformed cultures. Notably, the number of normal cotyledonary embryos in transgenic cultures was 3 times that in non-transformed cultures, indicating that LaCDKB1;2 not only increases the proliferation of cultures and the number of somatic embryos but also improves the quality of somatic embryos. These results provide insight into the regulatory mechanisms of somatic embryogenesis as well as new Larix breeding material.

miRNA-489-3p Improves Sepsis-Induced Lung Injury by Regulating Toll-Like Receptor 4 (TLR4) Signaling Pathway

Aim: To discuss miRNA-489-3p in sepsis-induced lung injury. Materials and methods: Using NR-8383 as research cell in our study, and using LPS stimulation to sepsis induced lung injury vitro model. Measuring cell proliferation by MTS assay; using Elisa assay to measure IL-1β, IL-6 and TNF-α concentration; apoptosis cell number and apoptosis rate were evaluated using TUNEL and flow cytometry; gene and protein were measured by RT-PCR and WB assay; measuring p-NF-κB(p65) nuclear volume by Immunofluorescence (IF); using Luciferase reporter assay to analysis miRNA-489-3p and TLR4 correlation. Results: Cell proliferation rate significantly down-regulated (P < 0.001), apoptosis cell number and apoptosis rate significantly increased (P < 0.001); IL-1β, IL-6 and TNF-α concentrations significantly up-regulated (P < 0.001); TLR4 and MyD88 gene and protein significantly increased (P < 0.001), NF-κB(p65) mRNA and p-NF-κB(p65) protein and nuclear volume significantly increased (P < 0.001). However, with miRNA-489-3p supplement, the cell proliferation rate, apoptosis cell number and apoptosis rate were significantly improved (P < 0.001, respectively) via TLR4, MyD88 and NF-κB(p65) mRNA and protein significantly depressing (P < 0.001, respectively). By LUC assay, miRNA-489-3p could target to TLR4 in NR-8383 cell. Conclusion: miRNA-489-3p overexpression had effect to improve sepsis induced lung injury via regulation TLR4/MyD88/NF-κB(p65).

Evidence for a Negative Correlation between Human Reactive Enamine-Imine Intermediate Deaminase A (RIDA) Activity and Cell Proliferation Rate: Role of Lysine Succinylation of RIDA

Reactive intermediate deaminase (Rid) proteins are enzymes conserved in all domains of life. UK114, a mammalian member of RidA subfamily, has been firstly identified as a component of liver perchloric acid-soluble proteins (L-PSP). Although still poorly defined, several functions have been attributed to the mammalian protein UK114/RIDA, including the reactive intermediate deamination activity. The expression of UK114/RIDA has been observed in some tumors, arousing interest in this protein as an evaluable tumor marker. However, other studies reported a negative correlation between UK114/RIDA expression, tumor differentiation degree and cell proliferation. This work addressed the question of UK114/RIDA expression in human non-tumor HEK293 cell lines and in some human tumor cell lines. Here we reported that human RIDA (hRIDA) was expressed in all the analyzed cell line and subjected to lysine (K-)succinylation. In HEK293, hRIDA K-succinylation was negatively correlated to the cell proliferation rate and was under the control of SIRT5. Moreover, K-succinylation clearly altered hRIDA quantification by immunoblotting, explaining, at least in part, some discrepancies about RIDA expression reported in previous studies. We found that hRIDA was able to deaminate reactive enamine-imine intermediates and that K-succinylation drastically reduced deaminase activity. As predicted by in silico analysis, the observed reduction of deaminase activity has been related to the drastic alterations of hRIDA structure inferred by K-succinylation. The role of hRIDA and the importance of its K-succinylation in cell metabolism, especially in cancer biology, have been discussed.

Health Informatics of Composite Nanoparticles as an Effective Contrast Agent in Magnetic Resonance Imaging and for Glioblastoma Radiosensitization

Background: Radiation therapy is a method using radiation to treat tumors, and has become commonly used and most effective treatments for malignant tumors. However, radiotherapy still has the shortcomings of high radiation dose, especially radiation resistance of tumor cells. Multifunctional nanoradiosensitizers can enhance the radiosensitivity of tumor cells and improving the effect of radiotherapy. Hyaluronic acid-functionalized ironbismuth composite particles are an effective contrast agent for magnetic resonance imaging (MRI), and can be used as radiosensitizers. Method: Hyaluronic acid-functionalized iron-bismuth composite nanoparticles HABilOPs can be prepared by a hydrothermal polyol method. In the cytotoxicity experiment, we took human glioblastoma cell line U87MG in logarithmic growth phase and rat vascular smooth muscle cells. We set different mass concentrations of 0, 15, 30, 45, 60, 100, and 300 mg/LHA-BMOPs was cultured for 48 h to calculate the cell proliferation rate. In the histocompatibility experiment, we injected HA-BilOPs solution into the tail vein of ICR mice to observe the pathological changes of mouse organs. In the radiosensitization experiment, we took U87MG cells in logarithmic growth phase for group culture. X-ray irradiation of 0, 3, 6, 9 Gy was given, and the HA-BilOPs group was added with culture medium of different mass concentration of LHA-BilOPs, and the combined group was added to the culture medium of different mass concentration of HA-BilOPs first, and then given X-ray irradiation of 0, 3, 6, 9 Gy. To analyse the features of glioblastoma, the conventional MRI manifestations of glioblastoma were examined. Results: We found that different concentrations of HA-BilOPs had no significant effect on the proliferation of vascular smooth muscle cells and U87MG cells. It was found that no pathological changes occurred in mouse organs after tail vein injection of HA-BilOPs solution. Nanoparticles can be phagocytosed by U87MG cells and it was found that U87MG cell proliferation rate had a significant linear negative correlation with HA-BilOPs mass concentration (0–200 mg/L) and radiation dose (0–9 Gy). At 6 Gy X Under irradiation, 200 mg/L HA-BilOPs reduced the cell proliferation rate to (41± 7)%. Using 100 mg/L HA-BilOPs and 6 Gy X-ray irradiation for colony formation experiments, the proliferation rate of U87MG cells in the combined group was statistically lower than that of the blank control group and radiotherapy group (P < 0.05). Conclusion: Hyaluronic acid-functionalized iron-bismuth composite nanoparticles have a significant radiosensitizing effect on glioblastoma and proved to be a good contrast agent used in MRI.

Bone Regeneration Using Rat-Derived Dedifferentiated Fat Cells Combined with Activated Platelet-Rich Plasma

Bone regeneration using mesenchymal stem cells has several limitations. We investigated adipose-derived dedifferentiated fat (DFAT) cells as an alternative, and evaluated their cell proliferation rate, osteoblast differentiation, and bone regeneration ability in combination with activated platelet-rich plasma (aPRP). Rat DFATs and aPRP were isolated using ceiling culture and centrifugation, respectively. The cell proliferation rate was measured, and the cells were cultured in an osteoblast differentiation medium under varying concentrations of aPRP for 21 days and stained with Alizarin red. Gene expression was evaluated using real time polymerase chain reaction. Critical defects were implanted with DFAT seeded gelatin sponges under aPRP, and four weeks later, the bone regeneration ability was evaluated using micro-computed tomography and hematoxylin-eosin staining. The cell proliferation rate was significantly increased by the addition of aPRP. Alizarin red staining was positive 21 days after the start of induction, with significantly higher Runt-related transcription factor 2 (Runx2) and osteocalcin (OCN) expression levels than those in the controls. A 9 mm critical defect was largely closed (60.6%) after four weeks of gelatin sponge implantation with DFAT and aPRP. Therefore, materials combining DFAT cells and aPRP may be an effective approach for bone regeneration. Further research is needed to explore the long-term effects of these materials.

PSIV-22 Supplemental effects of dietary lysophospholipids in lactation diets on sow performance, milk composition, and intestinal health of piglets

Dietary lysophospholipids could enhance nutrient utilization through a structural change of enterocyte membrane with increasing permeability. The objective of this study was to determine supplemental effects of dietary lysophospholipids in lactation diets on sow performance, milk characteristics, and intestinal health of piglets. The 52 pregnant sows were allotted to 2 treatments in randomized complete block design with parity and BW as blocks at d 110 of pregnancy. The treatments were CON (no added lysophospholipids) and LPL (at 0.05% lysophospholipids; Lipidol-Ultra, Pathway Intermediates, Shrewsbury, UK). The lactation diets were formulated to meet or exceed nutrient requirements suggested by NRC (2012). Milk samples from 12 sows per treatment were collected to measure gross energy, protein, fat, fatty acid profile, and immunoglobulins (IgG and IgA) on d 1 and d 18 of lactation. Twelve piglets per treatment were euthanized on d 18 to collect tissues to measure tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), malondialdehyde, protein carbonyl, IgA, microbiota in jejunal and colonic mucosa, morphology and crypt cell proliferation rate in the jejunum. Data were analyzed using the MIXED procedure of SAS. Sows fed LPL tended to increase (P = 0.084) litter size (11.9 vs. 12.6) on d 18 of lactation and decrease (P = 0.079) ADFI (8.72 vs. 8.02 kg) during d 9 to d 18 of lactation. Sows fed LPL tended to increase (P = 0.092) IgG (1.14 vs. 1.94 g/L) in the milk. Sows fed LPL increased (P &lt; 0.05) crypt cell proliferation rate (39.38 vs. 40.94%) in the jejunum. Supplementation of lysophospholipids in lactation diet did not affect proinflammatory cytokines, oxidative stress markers, and microbiota in jejunum and colon of piglets on d 18 of lactation. In conclusion, supplementation of dietary lysophopholipids improved productive performance and the intestinal cell proliferation of piglets with enhancing IgG concentration in the milk.